NABARD RESEARCH STUDY - 10

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FINAL REPORT OF NABARD PROJECT

Development of Iron-enriched Spent Hen Meat Products for

Boosting Layer Industry and Entrepreneurship

(14th May 2018 to 13th August 2020)

SUBMITTED BY

Dr. D SAPCOTA

Principal Investigator

College of Veterinary Science,

Assam Agricultural University

Khanapara, Guwahati-22

SUBMITTED TO

DEPARTMENT OF ECONOMIC ANALYSIS & RESEARCH (DEAR)

NABARD, BANDRA KURLA COMPLEX, BANDRA (EAST)

MUMBAI- 4000 51

About NABARD Research Study Series

The NABARD Research Study Series has been started to enable wider dissemination of research conducted/sponsored by NABARD on the thrust areas of Agriculture and Rural Development among researchers and stakeholders. ‘Development of Iron Enriched Spent Hen Meat Products for Boosting Layer Industry and Entrepreneurship’ is the tenth in the series. India reportedly tops the list of nations with most anaemic women and children. Iron deficiency is rampant among children below the age of three and women. This study looks at how meat of spent laying hens can be utilized in producing meat products fortified with heme iron through the usage of blood suiting the taste and convenience of school goers as well as their mothers. The study also proposes an economic model for preparing iron-enriched products from spent hens for small entrepreneurs. Complete list of studies is given on the last page.

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DISCLAIMER

This study has been supported by the National Bank for Agriculture and Rural Development (NABARD) under its Research and Development (R&D) Fund. The contents of this publication can be used for research and academic purposes only with due permission and acknowledgement. They should not be used for commercial purposes. NABARD does not hold any responsibility for the facts and figures contained in the book. The views are of the authors alone and should not be purported to be those of NABARD.

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TABLE OF CONTENTS

S. No. Chapters Page No.

Acknowledgement 4

Executive summary & Recommendation 5-8

1. Introduction 9-10

2. Review of literature 11-33

3. Objectives & Methodology 34-40

4. Results & Discussion 41-75

5. Summary & Conclusion 76-78

References 79-88

Annexure I: Score card for taste panel evaluation 89

Annexure II: Small scale preparation model scheme for

iron enriched chicken meatballs and nuggets

90-93

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ACKNOWLEDGEMENT

The research team of the project expresses its thankfulness to the NABARD, Mumbai for providing

financial support to carry out the research. The team is also thankful to the Director of Research

(Vety), AAU, Khanapara campus for providing necessary facilities to implement the project.

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EXECUTIVE SUMMARY & RECOMMENDATIONS

In the recent times, there has been an increase in the layer poultry production in the North-

East Region (NER), especially in the state of Assam. Around 70,000 eggers have been reported in

the state and the number is still growing as layer enterprise (News Bulletin Assam Vety. Council,

July-Dec/2016). The projected growth in layer industry is expected to be 2-3%, annually. The

surge in maize and oil seed production, of late, in the state must have boosted the layer industry to

achieve this feat which was initially non-existent. These birds after completion of their laying

cycle, termed as spent layers, are disposed off as culled birds at a very low-price (Rs. 30-50/kg) in

contrast to their broiler counterparts. The meats of these birds are tougher, less juicy and have

lower meat-to-bone ratio, thus, making it inferior in quality in comparison to meat of broilers. At

present, the availability of spent hen meat is sporadic which is limited to the area of production

and its market structure is not scientifically organized. It is envisaged that in near future more

number of layer farms will come up in the state which may result in the price of culled bird to

further dip due to low acceptance by the consumers and eventually create a negative impact on the

layer farming. Therefore, there is a need to develop certain technology so as to utilize the culled

birds to give a sustainable support to budding layer farmers.

On the other hand, there is a high demand for processed meat since more than 95 per cent

of the population is non-vegetarian in the NER region. Though there is no authentic data on the

quantum of processed meat (produced locally or imported from outside the state), the significant

trade in the supermarkets, KFC/J14 and other innumerable outlets indicate the growing

consumption and fondness of poultry meat by the meat lovers.

On the other hand, India reportedly tops the list of nations with most anaemic women

and children (IndiaSpend, Oct. 27/2016). Iron-deficiency (anaemia) in India is rampant among

children below the age of three (78.9 %) and women (55 %). In another report, it was recorded

that 72% of Indian married women are suffering from anaemia (Assam Tribune, Apr. 17/2017).

It has also appeared that the Nutritional Anaemia Prophylaxis Programme of issuing a weekly

iron table to adolescent girls and boys could not effectively solve the problem of anaemia.

The success of blood-fortified foods in addressing iron deficiency has been reported by

several researchers. Blood is a rich source of iron and proteins of high nutritional and functional

quality (Ofori and Hsieh, 2012). More utilization of slaughterhouse blood could prove

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nutritionally advantageous as blood is a rich source of heme iron (Oellingrath and Slinde, 1985)

and are more readily available than the non-heme type. It is expected that blood-fortified foods

may be readily accepted by the consumers judging from the eating habits of the people of the

NER region.

In this context, the present study strives to address the above stated problems

simultaneously, by utilizing the meat of spent laying hens in producing meat products fortified

with heme iron through the usage of blood suiting to the taste and convenience of school goers

as well as their mothers. The objectives of the study are as follows:

i. To encourage layer poultry farmers by utilizing their spent hens so as to run the

production cycle smoothly.

ii. To develop technology for preparation of iron-enriched processed meat products from

spent hens, suiting to the convenience of school going children and women.

iii. To study the quality characteristics of newly developed spent chicken products.

iv. To propose an economic model for preparing iron-enriched products from spent hens for

small entrepreneurs.

v. Transfer of technology to the end-user.

Healthy spent hens, quantum sufficit, were collected from layer egg producing farms,

scientifically slaughtered and processed. The carcass was deboned to collect their meat, along

with fat, preserved at -21°C until further use. The blood was collected during the slaughter to

which food grade anticoagulant Sodium citrate (citric acid) was added @ 0.3g/100ml to prevent

coagulation and was stored at refrigerated temperature (4±1°C) until further use. The meat was

minced using automatic mincer prior to the preparation of meat products. Several pilot studies

were conducted to evaluate the desirable and feasible levels of whole blood incorporation to the

final product. The basic recipes for both products were standardized and lean meat was replaced

with whole blood at 11%, 14% and 17% for chicken nuggets while for chicken meat balls, lean

meat was replaced with whole blood at 5%, 7.5 and 10% and prepared according to standard

method. The organoleptic quality, iron and cholesterol content, physico-chemical, proximate

composition were evaluated and storage stability studies (pH, WHC, TBA, Tyrosine value,

microbiological status) were conducted at an interval of 5 days up to 15 days for both the

products packaged under aerobic and vacuum condition and stored at 4±1°C. The data on all

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parameters were analyzed using appropriate statistical analytical methods to arrive at a valid

conclusion. Subsequently, ‘small-scale poultry product preparation model’ unit was developed

for the preparation of iron-enriched poultry products, i.e., meat balls and meat nuggets

commercially so as to encourage educated unemployed youths to attract towards starting

entrepreneurship

From the studies conducted it was found that the blood incorporated meat products

showed a significant (P<0.05) increase in the iron content to the extent of 154.56% in

meatballs and 264.72% in nugget at highest level on comparison to their control counterparts.

Simultaneously, the protein content in both the products were significantly (P<0.05)

increased. On the other hand, both fat and cholesterol contents in meatballs and nuggets were

significantly (P<0.05) reduced. On sensory evaluation of the chicken meatballs and chicken

nuggets, it was observed that an addition of whole blood up to 7.5% to chicken meatballs and

11% to chicken nuggets did not affect their overall acceptability wherein the panelist chose

‘Excellent to Very good’ score. This revealed the acceptability and possibility of using blood

at the said levels without any detrimental effect on sensory qualities. The meatballs and

nuggets retain their sensory qualities up to 15 days of storage at 4±1°C. The various storage

stability studies of the meat products were undertaken for refrigerated temperature (4±1°C) at

an interval of 5 days up to 15 days with aerobic and vacuum packaging module. The meat

products stored under room temperature showed spoilage, 24 hours post-preparation,

indicating room temperature as unfavorable for storage and unsafe for consumption.

The pH values of freshly prepared chicken meatballs and chicken nuggets increased

proportionately with the increase in the level of blood incorporation which might be due to

the influence of higher blood pH (7.6). During storage period at 4±1°C the pH value

gradually increased with the time, starting from the 0 day to 15th day, irrespective of the

blood level incorporated or the packaging methods. The thiobarbituric acid (TBA) values of

chicken meatballs and chicken nuggets were found to rise significantly (P<0.05) with the

increase in blood levels demonstrating a higher lipid oxidation than their control counterparts,

which may be due to the higher catalytic impact of iron on lipid oxidation. The TBA values

increased progressively with storage time. All the blood incorporated groups of chicken

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meatballs and nuggets showed significantly (P<0.05) higher tyrosine values which might be

due to the higher pH level and richer protein content in the blood. The tyrosine value

increased proportionately with the storage time in all the groups under both packaging system

owing to the intrinsic changes. The water holding capacity (WHC) of the chicken meatballs

and chicken nuggets were found to be significantly (P<0.05) higher in all the blood treated

groups. The replacement of lean meat with whole blood at increasing levels, higher pH and

strong emulsion resulted in elevated moisture level and subsequently higher WHC. However,

with increase in storage period, decrease in WHC was observed resulting from denaturation

of myofibrillar proteins and loosening up of muscle-microstructures allowing more water to

be entrained.

No microbial growth was observed in any of the samples till 5th day but was subsequentlynoted

with the extension of storage period. However, the total plate count (TPC) of all the groups were

within the permissible level (log 103 cfu/g of sample), both under aerobic and vacuum

packaging. Also, neither yeast and mould nor incidence of Salmonella were detected in both the

meat products (aerobic and vacuum packaged) throughout the storage period. It was interesting

to note that during the entire study period the vacuum packaging gave better results than its

aerobic counterpart, in terms of sensory qualities as well as physico-chemical and microbial

status. The findings of the study revealed that chicken meatballs with 7.5% and chicken nuggets

with 11% blood incorporation could increase the iron content to the extent of 99.62 and 184.49

%, respectively with remarkable protein elevation. Most importantly, these two levels were

highly acceptable in regard to sensory qualities.

The major issue observed in the study was shelf life of the meat products under

refrigeration temperature (4±1°C). The meat products remained safe for consumption without

any affect in the quality up to 10 days. It is recommended that the meat products may be stored

under freezing temperature (18-21°C) for better shelf life and longer storage duration (3

months). Also, vacuum packaging is recommended over aerobic packaging as the vacuum

packaging gave better results than its aerobic counterpart, in terms of sensory qualities as well as

physico-chemical and microbial status.

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CHAPTER I

INTRODUCTION

The state of Assam has witnessed a rise in the number of layer farms established in the recent

times with a steady growth in this sector. It has been reported that the Northeast Region of India

imports around 42 lakh of table eggs from outside the state, daily. Out of which Assam gets around 70

per cent (29.40 lakh in number). Thus, there is a high demand of table eggs in the state. Of late, it has

been observed that maize cultivation in the state has exponentially been increased, which may be due

to the high demand of this feedstuff as cash crop and also as animal feed. On the other hand, certain

companies, like M/s Godrej, Sibasais Oils etc., have taken up palm oil seeds cultivation, covering

large areas of Assam (especially, Goalpara, Kamrup and Bongaigaon districts), Arunachal Pradesh,

Nagaland and Mizoram. Both these feeds are the high sources of energy in the diet for poultry.

Further, other private companies have started producing DDGS (Dried distillers grain soluble)

industrially, mostly in the districts of Kamrup (Saigaon) and Karbi Anglong (Khatkhati), which has

given a boost to the protein deficient poultry feed industry. Needless to say that Assam is self

sufficient on rice polish production. Thus, the entire scenario is changing to favour a leaping progress

for layer poultry production in the state.

Conventionally, the layer chickens, after completion of their 72 weeks economic

production period in the commercial farms are sold as spent hens/culled birds. Being matured birds,

their meat are tough, less tender/juicy, making it inferior in quality in comparison to meat of broilers

and therefore less preferred by the consumers,. They are sold at a very low-price, around Rs. 30-50/kg

which is a stark contrast to their broiler counterparts. Due to this problem often-time it becomes

difficult to sell off these birds. At present, the availability of spent hen meat is sporadic which is

limited to the area of production and its market structure is not scientifically organized. It can be

foreseen that in near future large numbers of layer farms will come up in the state and the price of

culled bird may further dip due to low acceptance by the consumers, eventually giving a negative

impact on the layer farming. Therefore, there is a need to develop certain technology so as to utilize

the culled birds to give a sustainable support to budding layer farming.

Additionally, there is a high demand for processed meat since more than 95 per cent of the

population is non-vegetarian in the NER region. Though there is no authentic data on the quantum of

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processed meat (produced locally or imported from outside the state), the significant trade in the

supermarkets, KFC/J14 and other innumerable outlets indicate the growing consumption and fondness

of poultry meat by the meat lovers.

Recently, India has witnessed unprecedented incident of Covid-19 cases due to which large

number of people hailing from Assam who were engaged in private sectors in other states have

returned to the state. The prevailing uncertainty to resume their original work place has left, many of

them preferring to work in their mother state. Since poultry farming is relatively an easier avocation to

try for, many have shown interest as entrepreneurship. Parallelly, the state government is also

expending various schemes so as to help the returned migrant workers. These schemes carries soft

loans with government subsidies. Needless to say that this opportunity will give further fillip to the

upcoming poultry industry.

Deficiency of iron in human body is the most common nutritional deficiency prevalent

worldwide and more extensive among women and children. India reportedly tops the list of nations

with most anaemic women and children (IndiaSpend, Oct. 27 2016). In the country, the iron-

deficiency, anaemia was reported in 58.6% of children, 53.2% of non-pregnant women and 50.4% of

pregnant women in 2016, as per the NFHS (National Family Health Survey). India carries the highest

burden of the disease despite having an anaemia control programmes for the last 50 years. In another

report it was observed that 72% of Indian married women suffer from anaemia (Assam Tribune, Apr.

17, 2017). It has also appeared that the Nutritional Anaemia Prophylaxis Programme of issuing a

weekly iron table to adolescent girls and boys could not be able to solve the problem of anaemia.

The success of blood-fortified foods in addressing iron deficiency has been reported by several

researchers. Blood is a rich source of iron and proteins of high nutritional and functional quality (Ofori

and Hsieh, 2012). More utilization of slaughterhouse blood could prove nutritionally advantageous as

blood is a rich source of heme iron (Oellingrath and Slinde, 1985) and are more readily available than

the non-heme type. It is expected that blood-fortified foods may be readily accepted by the consumers

judging from the eating habits of the people of the NER region.

In this context, the present study strives to address the above stated problems simultaneously,

by utilizing the meat of spent laying hens in producing meat products fortified with heme iron through

the usage of blood suiting to the taste and convenience of school goers as well as their mothers.

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CHAPTER II

REVIEW OF LITERATURE

A) Meat balls

Grasso et al. (2019) investigated the effect of texturized soy protein (TSP) at different inclusion levels

(15% and 30%) with and without nutritional yeast as flavour enhancer on the sensory and instrumental

texture quality of beef meatballs, compared to a soy and yeast-free control. Sixty participants assessed

the samples using Check-all-that-apply (CATA) questions and hedonic scales. Overall, the texture of all

TSP-containing samples received significantly higher acceptability scores than control, while 15% TSP

with yeast received the highest flavour and overall acceptability scores. Penalty-lift analysis of CATA

terms identified the main drivers for liking as ‘‘moist looking’’, ‘‘juicy’’, ‘‘soft’’ and ‘‘crumbly and

easy to cut’’. Control samples were significantly more often associated than the other recipes to the term

‘‘hard’’, a key driver for dislike and the least associated to ‘‘soft’’ and ‘‘crumbly and easy to cut’’.

Adding 15–30% TSP with or without yeast inclusion could be beneficial for the development of future

meat hybrids with acceptable sensory quality.

Minantyo et al. (2019) studied the effect of adding Kelor (Moringa Oleifera Lam) leaves on the

nutritional and organoleptic characteristics of milkfish meatball. The aim of the present work was to

improve the protein, calcium, and fibre contents and organoleptic characteristics of milkfish (Chanos

chanos) meatballs. Results obtained showed that calcium and fibre content of the meatball was

increased which may be due to presence of vital minerals (calcium, copper, iron, potassium,

magnesium, manganese and zinc) and natural antioxidants present in kelor leaves. However, there was a

reduction in the protein content of milkfish meatballs. Milkfish meatballs added with 10% boiled kelor

leaves were most accepted by the panelists.

Çağlar et al. (2018) investigated the effect of yellow, black, and brown mustard seeds on color,

thiobarbituric acid-reactive substances (TBARS, microbiological and sensory qualities of meatballs

during storage. Heat treatment of mustard seeds decreased the TBARS value of meatball samples

(p<0.0001). The addition of mustard seeds decreased count of aerobic mesophilic bacteria (p<0.0001),

enterobacteriaceae (p<0.0001), psychrophilic bacteria (p<0.0001) and yeast and mould of the meatball

samples (p<0.0001). On 15th day of storage, the yellow mustard added meatballs sample showed better

colour, appearance, flavour and acceptability ratings than those added with black and brown mustard

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seeds. The improvement in microbiological quality of the meatballs was attributed to the antibacterial

and antifungal properties of the mustard seeds.

Essa et al. (2018) assessed the influence of sugar beet pulp powder on reduction of fat when added to

meatballs. The fat in the meatball was replaced with sugar beet pulp powder at the ratio of 25 and 50 %

of animal fat. Chemical composition, oil holding capacity and water holding capacity (WHC) of raw

materials were examined and the meatballs were estimated of cooking attributes and sensory

characteristics. The cooking yield of meatballs was reported to be increased while cooking loss

decreased which might be attributable to the ability of sugar beet pulp powder to hold excess water. No

significant difference (p ≥ 0.05) in sensory properties was found between control and the sugar beet pulp

powder added meatballs.

Jahan et al. (2018) conducted an experiment to find out the effect of different levels of pomegranate

extract and synthetic antioxidant (Beta Hydroxyl Anisole) on the quality of beef meatballs. Pomegranate

extract at 0.1%, 0.2%, and 0.3% were added to fresh and preserved beef meatballs and stored for 60

days which were evaluated for proximate composition, sensory, physicochemical, biochemical and

microbiological quality. The experiment was conducted by factorial experiment in completely

randomized design. CP, EE, raw and cooked pH value, sensory traits significantly increased (p<0.05)

while DM, Ash, FFA, POV, TBARS, TVC, TCC, TYMC decreased (p<0.05) in all treatments. On the

other hand, significant decrease (p<0.05) in all the sensory traits, CP, raw and cooked pH, cooking loss,

TCC, and TYMC at different days of intervals were reported. In view of the sensory, nutritional,

physicochemical, biochemical and microbial properties, 0.3% pomegranate extract was recommended

for value addition in beef meatball which enriches the product with natural antioxidant.

In a study, Yeung and Huang (2018) explored the effects of food proteins-soy protein, sodium caseinate,

whey protein, egg albumen powder, and skim milk powder on the texture and sensory acceptability of

phosphate-free meatballs. The results were compared with the texture and sensory acceptability of

meatballs with phosphate. Textural studies revealed that adding of different food proteins to phosphate-

free meatballs increased its hardness and the hardness caused by egg albumen powder was recorded

highest, followed by sodium caseinate and whey protein. Phosphate-added and phosphate-free meatballs

without any protein additives were reported to be less chewy than meatballs with protein additives.

Addition of egg albumen powder, sodium caseinate, and whey protein, however, could increase the

meatballs’ chewiness. The pH values and water compositions of all groups of meatballs were recorded

between 6.36 and 6.96 and between 49.81% and 56.79%, respectively. Sensory evaluation revealed that

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soy protein had negative effects on sensory acceptance, whereas whey protein scored the highest in

overall sensory evaluation. The hygiene quality test showed that the total plate count of all meatball

groups had less than 105 CFU/mL while Escherichia coli and coliforms were not detected.

Bayomy et al. (2017) reported that the quail meatballs and pickled quail eggs as market products were

acceptable by the consumers. The nutritional composition of the quail pickled eggs and quail meatballs

were also determined. The quail meatballs showed 70.56% moisture content and 29.44% dry matter

which included 7.78%, 12.98%, 7.10% and 2.48% for total lipids, proteins, ash and carbohydrate,

respectively on wet basis. Total cholesterol content in quail meatballs was recorded at 0.087g/100g. The

nutritional values of pickled quail eggs recorded for moisture, dry matter, total lipids, proteins, ash and

carbohydrate on wet basis were 53.32%, 46.68%, 20.09%, 20.38%, 4.70% and 1.51%, respectively. The

average cholesterol content in pickled quail eggs was 2.06 g/100 g yolk. Total essential and non-

essential amino acids were 6.17g/100g and 5.87g/100g protein, respectively in quail meatballs. In

pickled quail eggs, the values of total essential amino acids and nonessential amino acids were recorded

at 6.84g/100g and 8.81g/100g protein, respectively. Total saturated fatty acids (SFAs) in quail meatballs

and quail pickled eggs were 26.37% and 37.52%, respectively. Total mono-unsaturated fatty acids and

poly-unsaturated fatty acids contents in quail meatballs were 36.61% and 36.21, respectively, while

these values for pickled quail eggs were recorded at 54.12% and 8.36%, respectively.

Okuskhanova et al. (2017) presented a new technology with poultry meat (neck and back part), rice, sea

cabbage (Laminaria) and carrot. Three variants of meatball were prepared with different weight ratios of

Laminaria i.e., Variant 1 (15%), Variant 2 (10%) and Variant 3 (5%). The comparative quality and

organoleptic indicators of meatballs were studied. It was noted that the developed meatballs had soft

consistency, pleasant flavour, better sensory characteristics and balanced composition. The highest

protein level was recorded in variant 2 (19.7%) while the lowest was observed in the control group

(10.1%). Variant 2 meatballs also showed an increased level of mineral elements (3.11%) in comparison

to variant 1 (2.6%) and variant 3 (1.6%). The moisture and fat content of the developed meatballs

showed higher values than the control group. Different proportions of Laminaria in meatball

formulations caused significant changes in content of I, Mg, K and Na. Reduction in the concentrations

of these elements were observed when the Laminaria weight ratio in meatballs was lowered which

points to the positive effect of Laminaria on food quality.

Ferawati et al. (2017) evaluated the microbiological quality and safety of meatballs collected from five

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different manufacturers around Payakumbuh City, West Sumatra, Indonesia. Microbiological analysis

of meatball samples resulted in aerobic plate count range from 7 log CFU/gr to 8.623 log CFU/gr. Total

coliform ranges from 1.041 log Most Probable Number (MPN)/gr to 3.380 log MPN/gr. The qualitative

detection of borax and formalin content on all meatball samples were found negative. Thus, it remains

essential to include the significance of effective hygiene practices as an important safety measure in

consumer education programmes.

The addition of coriander extracts to turkey meatballs was studied to determine its effects on the quality

of the meatballs (Gantner et al., 2017). The meatballs were stored in modified atmosphere at 4°C±1°C

for 9 days. The addition of coriander extract at 500 ppm level delayed the process of lipid oxidation

during 6 days storage period and growth of aerobic microorganisms was inhibited up to 9 days of

storage. The usage of dose at 500 ppm as well as at 200 ppm had no significant effect on the sensory

features, but had an impact on colour parameters of the meatballs. In all the meatballs added with

coriander extract, volatile terpenes were identified.

Ozturk et al. (2017) applied the vacuum cooling and the conventional cooling techniques for the cooling

of the meatball and to show the vacuum pressure effect on the cooling time, the temperature decrease

and microbial growth rate. The results of the vacuum cooling and the conventional cooling (cooling in

the refrigerator) were compared with each other for different temperatures. The study shows that the

conventional cooling was much slower than the vacuum cooling. Moreover, the microbial growth rate of

the vacuum cooling was extremely low compared with the conventional cooling. Thus, the lowest

microbial growth occurred at 0.7 kPa and the highest microbial growth was observed at 1.5 kPa for the

vacuum cooling. The mass loss ratio for the conventional cooling and vacuum cooling was about 5%

and 9% respectively.

Reddy and Vani (2017) selected sorghum, oat and barley flours each at 5, 10 and 15 percentages to

prepare value added chicken meat balls. Among different treatments, chicken meat balls with 15% oat

flour produced significantly (p<0.05) higher cooking yield, emulsion stability, water holding capacity,

penetration values, % crude fibre, moisture % with low crude fat % and better organoleptic traits than

the rest of the formulations. Hence incorporation of oat flour at 15% level was considered to be

optimum with all the desired qualities of value added chicken meat balls.

Zamora et al. (2017) utilized breadnut as a meat replacement for meatball to determine the most

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preferred formulation of breadnut meatball, sensory acceptability, cost of direct materials and shelf life

of breadnut meatball. The three formulations of the product were: Formulation 1- 25% breadnut and

75% ground beef, Formulation 2- 50% breadnut and 50% ground beef and Formulation 3- 75%

breadnut and 25% ground beef. To determine the acceptability of the product, consumer type panel was

used. It was noted that Formulation 1 was the most preferred group. Based on the sensory acceptability

of 60 respondents, there was no significant difference among the formulated products in terms of

general acceptability, texture, flavour and after taste. The costs of direct materials for formulated

products were also cheaper than the control group per serving of 43 grams + 2 grams. The shelf life of

the formulated product was two weeks from the day it was formulated.

Deepshikha Deuri et al.(2016) developed a ready-to-eat Vawksa rep (smoked pork product) and studied

the synergistic effect of curing ingredients and vacuum packaging on the physico-chemical and storage

quality during refrigerated storage at (4°C±1°C) for 15 days. Four different batches of Vawksa rep

samples were prepared, i.e., T-1 (uncured, first cooked at 121°C for 15 min, and then smoked at 120°C

for 30 min), T-2 (uncured, cooked, and smoked simultaneously at 120°C for 45 min), T-3 (cured, first

cooked at 121°C for 15 min, and then smoked at 120°C for 30 min), and T-4 (cured, cooked, and

smoked simultaneously at 120°C for 45 min).Cooking yield was significantly higher (p<0.05) for the T-

4. The pH of T-3 and T-4 samples were significantly higher (p<0.05) on day 15. The tyrosine value of

all the samples increased significantly (p<0.05) at the different time interval of the storage period.

Thiobarbituric acid value was significantly (p<0.05) lower in T-3 sample both at the beginning and at

the end of storage period. The microbiological profile reflected lower total plate count in T-3 and T-4

than the remaining group. In addition, Escherichia coli count was negative for T-3 and T-4 samples

throughout the storage period. The sensory attributes of T-3 and T-4 samples recorded superior scores

for colour, flavour, texture, juiciness, and overall acceptability.

Nasrin et al. (2016) carried out a study to determine appropriate level of wheat flour with bottle gourd

leaves extract which could be used in meatball production. Meatballs were prepared as per the

formulations categorized into four treatment groups viz., T1 (control), T2- (5% bottle gourd leaves

extract + 5% wheat flour), T3 (5% bottle gourd leaves extract+ 8% wheat flour ) and T4 (5% bottle

gourd leaves extract +10% wheat flour). The meatballs were packed under aerobic condition and stored

at refrigeration temperature. With the progression of the storage period, increase in the DM content was

reported in all the treatment groups. On the other hand, Crude protein, ether extract, raw and cooked pH

decreased (p<0.05) with storage period. Significant differences (p<0.05) were observed among the

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different treatment groups for colour, cooking loss, flavour, tenderness and overall acceptability.

TBRAS value, total viable count, total coli form count and yeast-mould count of meatballs showed

significant differences (p< 0.05) among the different treatment groups. The meatball group with 5%

bottle gourd leaves extract and 5% wheat flour was observed to be most acceptable than the other

treatment groups. Incorporation of wheat flour and bottle gourd leaves extract at 5% each can

successfully be used in manufacturing of meatballs with the added functional properties of wheat flour

as a prebiotic and antioxidant activity of bottle gourd leaves extract.

Verma et al. (2016) explored the possibilities of utilization of green cabbage (Brassica olerecea) in the

preparation of chicken meatballs. The green cabbage was incorporated at various levels, namely 0%,

15% and 25% by replacing lean meat in the formulation and subsequently storage at 4 ± 1°C. Crude

protein, fat, ash and energy content of the chicken meatballs showed significantly decreasing (p<0.05)

trend with increasing levels of green cabbage. There was a significant increase (p<0.05) in the moisture

content, cooking yield, moisture retention and moisture protein ratio in treated products than the control

and recorded highest for the group with 15% green cabbage. Chicken meatballs were aerobically packed

and assessed for storage quality under refrigerated (4 ±1°C) conditions. Sensory attributes showed a

significantly (p<0.05) decreasing trend for all the groups, whereas pH, total plate counts, yeast and

mould count, free fatty acid and thiobarbituric acid reacting substances values increased significantly

(p<0.05) throughout the storage period. The addition of green cabbage as lean meat replacement at 15%

could be used successfully in chicken meatballs conspicuous of the extended shelf life resulting from

retardation of the lipid oxidation by the antioxidant compounds present in green cabbage and the slowed

microbial growth under refrigerated (4 ±1°C) conditions.

Anandh (2015) prepared chicken meat balls stored at 4 ±1ºC to assess the quality changes after every

five days up to 20 day of storage. The pH, moisture content, thiobarbituric acid and tyrosine values

increased gradually while extract release volume decreased significantly with the increasing storage

period. Throughout the storage period, all microbial counts were recorded within the acceptable limits

of cooked meat products. Sensory scores decreased significantly after 15 days of storage. The findings

showed that chicken meat balls can be safely stored up to 15 days at 4 ±1ºC in LDPE pouches under

aerobic packaging.

Öpi and Harun (2015) investigated the interaction between cooking time and storage temperatures of the

processed chicken meatballs made from minced chicken meat vide the sous method. The chicken

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meatballs were categorised into four experimental groups according to the application of heat treatment

(10 and 20 minutes) and storage time (+2 and +10°C). Cl. perfringens and Listeria spp. were not

detected in meatball samples during the storage period. Samples cooked at 10 minute and stored at +2°C

showed the highest count of total aerobic mesophilic bacteria (p<0.05). Higher TBARS values for

samples stored at 10 °C were observed indicating higher lipid oxidation and suggest the evidence of

interaction between cooking time and storage temperature. On the contrary, cooking time and storage

temperature showed no influence on the colour and textural properties on the meatballs. From the study,

it was reported that cooking time x storage temperature had more affect on the microbiological and

chemical properties than the colour and textural properties of chicken meatballs.

Evivie et al. (2015) evaluated the organoleptic properties of soy meatball on inclusion of varying levels

of Moringa oleifera leaves powder. The result of the sensory evaluation of the products showed that the

soy meatballs were generally accepted above an average of 3.5. However, soy meatball sample without

Moringa oleifera leaves powder inclusion was most acceptable (very much liked) in terms of colour,

taste, texture and overall acceptance with a mean score of 4.37. The colour scores differed significantly

(P>0.05). The taste significantly (P>0.05) decreased with the increase in Moringa oleifera leaves

powder levels.

Ergezer et al. (2014) determined the effects of using different amounts of potato puree (PP) (10 and

20%) and 10% bread crumbs (BC) as an extender on chemical composition, energy values, colour

measurements, water holding capacity (WHC), penetration values, thiobarbituric acid value (TBA) and

sensory analyses of meatballs. The control group (C) were without extender. Meatballs were cooked in a

pre-heated 180°C electric oven. Uncooked meatballs formulated with 20% PP had the highest moisture

content. No significant differences were recorded for protein contents of uncooked meatballs. 20% PP

increased moisture and fat retention values and water holding capacity of meatballs. Meatballs with

10% BC had the lowest (the hardness in the texture) and meatballs with the 20% PP had the highest (the

softness in the texture) penetration values. Formulating meatballs at a level of 20% resulted lower L*

values. TBA values of control group were higher than PP added meatballs at the end of the storage

period. Flavour scores for meatballs formulated with PP were higher than control and meatballs

formulated with BC. Meatballs formulated with 10% PP had similar overall acceptability with meatballs

added with 10% BC.

Turhan et al.(2014) studied the nutritional and storage quality of meatballs formulated with different

levels of bee pollen (0, 1.5, 3.0, 4.5 and 6.0%) during storage at 41℃ for 9 d. Protein content of

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meatballs increased, while moisture content decreased with increased pollen. The addition of pollen

improved cooking loss but decreased the redness (Hunter a value) and sensory scores. Textural

properties (hardness, springiness, gumminess, and chewiness) were affected by pollen addition and the

hardness and gumminess values of meatballs decreased as the pollen content increased. While C18:0

content of meatballs slightly decreased with pollen addition, C18:2n-6c, C18:3n-3, C20:5n-3 and PUFA

contents increased. The PUFA/saturated fatty acids (P/S) ratio increased from 0.05 in the control to 0.09

in meatballs with 6.0% pollen. The n-6/n-3 ratio decreased from 11.84 in the control to 3.65 in the

meatballs with 6.0% pollen. The addition of pollen retarded the lipid oxidation and inhibited the

bacterial growth in meatballs. The values of pH, redness, TBA, total aerobic mesophilic bacteria,

coliform bacteria and S. aureus counts changed significantly during storage. The results suggested that

bee pollen could be added to enhance the nutritional and storage quality of meatballs with minimal

changes in composition and/or sensory properties.

Halini et.al. (2013) determined the effects on the quality of meatball prepared with different ratios of

chicken and duck meat. Meatballs were produced by using 5 different ratios of duck and chicken meat

i.e., A = 0:100, B = 25:75, C = 50:50, D = 75:25, and E = 100:0. Increase in the ratio of duck meat

caused increase in moisture, ash, and carbohydrate (by difference), but decreases in protein and fat. The

range of moisture, protein, fat, ash and carbohydrate content of meat ball in this study were 68.26 –

73.42 %, 10.45 – 5.88 %, 10.88 – 8.50 %, 1.84 – 2.86 %, 8.56 – 10.99 %, respectively. Determination of

texture and colour by instrument showed significant differences (P<0.05) among samples. The highest L

value (lightness) (65.89) was observed in formulation A which contained 100% chicken meat, while the

lowest L value (55.65) was observed in formulation E containing 100% duck meat. The addition of duck

meat thus contributed to a darker colour product. Texture analysis showed decreasing values of shear

stress (253.31 – 541.50) with the increase of duck meat ratio.

Bhat et al. (2013) evaluated the utilization of skin in the preparation of meat balls incorporating at

different levels viz., 25%, 50%, 75% and 100% replacing lean meat in the formulation. The meat balls

were further enrobed to see the effect of coating on the quality characteristics of meat balls. Both coated

as well as control meat balls were aerobically packed in low density polyethylene (LDPE) pouches and

stored at refrigerated condition at 4±1°C. The studies for various quality traits were conducted at a

regular interval of 0, 7 and 14 days. Significant reductions (P<0.05) were observed in the emulsion

stability, cooking yield, proximate composition and sensory attributes of meat balls with the increasing

skin level in the meatballs. Based on sensory studies, meat balls containing 50% skin were optimized as

best among the coated as well as non-coated meat balls. TBARS value, total plate count and

19

psychrophilic count increased significantly (P<0.05) whereas the scores for various sensory attributes

decreased significantly (P<0.05) with the progression of storage days. Coliforms were not detected

during the 14 days storage period. Meat balls utilizing chicken skin could be stored for a period of 14

days at refrigerated temperature (4±1°C) with slight changes in the quality attributes under acceptable

limits.

Ferawati and Marlida (2012) tested the effectiveness of using coconut shell liquid smoke in preservation

of meatballs at the concentration of 0%, 3%, 5% and & 7% and its effect on the shelf life at refrigeration

temperature on 0, 5, 10 and 15 days. Parameters like water content, protein content, fat content, pH,

Total plate count shelf life were found to be significant on protein content only. Application of 7%

liquid smoke on meatballs stored at 4±1o C were more observed to be most acceptable and its shelf life

increased up to 15 days storage with decreased in pH and moisture content in comparison to control.

The study concluded that liquid smoke could be used as an effective preservative agent for meatballs

and most preferably at 7%.

Several trial were conducted by Chandralekha et al. (2012) for incorporating the suitable binder at right

percentage for the development of value added chicken meat balls. Three different binders viz., Bengal

gram flour, corn flour and soy flour each at three different levels viz., 5, 10 and 15 percentages were

added to prepare the value added chicken meat balls to asses the binder at its optimum desirable level.

Among different treatments, chicken meat balls incorporated with Bengal gram flour at 10 percent level

recorded a significantly (p<0.05) higher yield, emulsion stability and higher water holding capacity and

better organoleptic traits than the rest of the formulations. Hence incorporation of Bengal gram flour at

10 percent level in chicken meat balls was considered to be optimum for all the desired qualities of a

value added meat product.

Gök and Bor (2012) investigated the effects of olive leaf extract (500 and 1000 ppm), blueberry extract

(500 and 1000 ppm) and Zizyphus jujuba extract (500 and 1000 ppm) on color, lipid oxidation

(thiobarbituric acid-reactive substances, TBARS) and microbiological and sensory quality of meatball

during storage periods. The addition of olive leaf extract, blueberry extract and Zizyphus jujuba extract

reduced (P < 0.001) TBARS in meatball. Olive leaf extracts showed higher antioxidant activity than

blueberry and Z. jujuba extracts. Samples having 1000 ppm of blueberry extract had the highest a*

value (P < 0.001). On a given storage day, samples having Z. jujuba extracts showed lower microbial

microbial count than other samples, indicating the antibacterial effect of Z. jujube. The highest scores (P

20

< 0.001) for overall sensory quality were recorded for the meatballs with 1000 ppm of blueberry fruit

extract.

Ikhlas, et al. (2011) analysed the proximate composition and physicochemical properties of quail

meatballs prepared with different types of flour. Meatballs were produced using 65% quail meat, 3%

flour (cassava, corn, wheat, sago and potato flour), 3.2% soy protein isolate, 10% palm oil, 2.1% salt,

2% sugar, 0.9% mixed spices and 13.8% cool water. The proximate composition of the quail meatballs

produced comprised of 64.94-66.33% moisture, 13.43-14.47% protein, 10.32-13.77% fat, 2.30-2.95%

ash and 4.80-7.67% carbohydrates. The analysed texture profiles were significantly different (p<0.05)

among all the treatment groups. The hardness of the quail meatballs using potato flour was recorded

highest (10.08 kg), followed wheat (9.18 kg), corn (9.08 kg), sago (8.45 kg) and cassava flour (7.90 kg).

The sensory evaluation of the quail meatballs produced a moderate score of 5 on a 7-point hedonic

scale. The sensory score showed that quail meat can be successfully used in the manufacture of

meatballs as an alternative to other meats such as beef and chicken, using different types of flour.

Cassava flour was found to be the best and most acceptable formulations for quail meatballs production.

Temelli et al. (2011) investigated the microbiological changes occurring during the processing stages of

chicken kadinbudu meatballs produced in a private poultry meat processing plant, Bursa/Turkey. One

hundred and seventy samples were collected from the production stages and non-meat ingredients were

examined for total aerobic mesophilic bacteria (TAMB), coliform bacteria, Escherichia coli (E. coli),

Enterobacteriaceae, enterococci, staphylococci-micrococci, coagulase positive staphylococci, yeast and

molds. In chicken ground meat, the TAMB, Enterobacteriaceae, enterococci, staphylococci-micrococci,

yeast and mold counts were recorded at 3.75, 2.38, 2.92, 2.77, 2.59 and 3.23 log cfu/g, respectively.

There was a significant increase in TAMB counts in samples of pre-dusted, battered and breaded patties

(p<0.05). Of the non-meat ingredients, only flour had a significant effect on the TAMB increase in pre-

dusted patty samples (R2 = 0.55, Beta = 0.74). There was approximately one log reduction in all sample

counts after frying, with statistically significant reductions only in TAMB and Enterobacteriaceae

counts (p<0.05). Samples after cooling and packaging had microbial counts under detection levels,

indicating good personnel hygiene, and cleaning and disinfection applications leading to zero post

contamination to the product after the cooking stage. E. coli or coagulase positive staphylococcus was

not detected in any of the samples either from the production stages or the non-meat ingredients. The

results indicated that neither the ground chicken meat nor the non-meat ingredients caused initial and

21

secondary and/or cross-contaminations for the final chicken kadınbudu meatballs. Furthermore, the heat

application with two stages was found to be sufficient for the production of a non-hazardous product

and there no post-contaminations after cooking were observed.

Siriken et al. (2009) studied a total of 100 Turkish sucuk samples for pysico-chemical and

microbiological quality. The maximum levels of moisture, fat and pH permitted according to the

standards of Turkish Food Regulations are 40, 40% Fresh Matter (FM) and 5.8, respectively and sucuk

should not contain any starch. In these respects, 51.5% and 32% of the samples was found high for

moisture and pH values, respectively and the fat value of all the samples were in line with the standard.

Interestingly, starch was detected in 66% of the samples. Lactobacilli and micrococci/staphylococci

were recorded at <102-108 and <102-107 cfu g-1, respectively. Negative correlations were observed

between Lactobacilli levels and pH values (p<0.01). However, such correlation was not observed in few

sucuk samples. NaCl was detected above 5% Dry Matter (DM) of 25 samples. Maximum and minimum

collagen and hydroxyproline levels were recorded at 3.20-9.68 and 0.40-1.21 μg mg-1 (DM),

respectively. The percentage of moisture, fat, pH, NaCl, starch, collagen and hydroxyproline levels

showed variation among the samples analysed. There is no official regulation for collagen and

hydroxyproline levels of Turkish sucuk. Therefore, these studies may serve as the base for the

preparation of Official Standards of quality for collagen and hydroxyproline content of sucuk in Turkey.

Vural and Aksu (2006) examined the effects of gamma irradiation on the microbiological and

physiochemical characteristics of meatballs. Meatball samples were produced experimentally and

divided into six subgroups. The first group (control group) was not subjected to an irradiation process

and the remaining groups were subjected to irradiation at doses of 1, 2, 3, 5 and 7 kGy. All groups were

analysed on 0 (the day of irradiation process), 5, 10, 15, 20 and 30 days. The study revealed a significant

reduction in the microbiological flora of the meatballs in relation to gradual increases in irradiation

dosage. Faecal coliform bacteria counts, coliform bacteria counts and S. aureus counts were eliminated

after applying 2 kGy, 3 kGy and 7 kGy irradiation doses, respectively. During storage at 4±1 °C, the

bacterial populations systematically increased. Compared to the control group, low dose irradiation did

not significantly change pH values, but higher pH levels were determined during high doses of

irradiation. Irradiation increased the peroxide levels, but this increase did not seem to correlate with

irradiation dosage. FFA levels decreased with increasing levels of irradiation dosage.

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Reddy et al. (2005) studied the effect of ginger added to chicken meat balls at 2, 4 and 6% levels,

divided into two batches i.e., deep fat fried and ready-to-eat (RTE) and the other unfried was ready-to-

cook (RTC). The chicken meat balls were packed in polyethylene bags, stored at -20±2°C and evaluated

at 15 days intervals up to 60 days. Increasing level of ginger resulted in higher pH values and flavour

scores and a decrease in thiobarbituric acid (TBA) values of chicken meat balls next to BHA added

samples. Statistically no significant effect was observed due to addition of ginger on penetrometre

values and microbial load. During frozen storage for 60 days, there was significant (p<0.01) increase in

pH and TBA values and decrease in penetrometer values, microbial load, flavour and juiciness scores.

RTC was found better than RTE samples for all parameters except microbial load. The results showed

that ginger at 6% level exhibited antioxidant property along with enhancement of flavour and juiciness

of the chicken meat balls.

Yılmaz et al. (2005) studied the effects of different cooking processes (grilling, oven and microwave

cooking) on the microbial flora of the raw meatballs inoculated with E. Coli O157:H7 at the level 2_104

cfu/ml stored at 4 _C. While Salmonella was found in each sample, none of the samples contained C.

perfringens or E. coli O157:H7. The processes of grill cooking or microwave cooking decreased the

microbial flora by 2–3 log cycles. E. coli O157:H7 was completely destroyed by all cooking methods.

E. coli O157:H7 count of the raw meatball samples increased by 1.5 log cycles at the end of storage

compared to beginning.

Salih et al. (1987) conducted a test for monitoring lipid oxidation in poultry products. Thiobarbituric

acid (TBA) values of cooked poultry meat were determined by a distillation method and by a modified

extraction method. Perchloric acid was used in the extraction method to release TBA reactive

substances from the meat. The maximum absorption wavelength of the TBA-malonaldehyde complex

was 531 nm. The TBA values for cooked chicken and turkey were highly correlated (r= .91) for both the

extraction and distillation methods. Sensory scores of warmed-over flavor in pre-cooked poultry were

highly correlated with TBA values in both the distillation (r= .83) and extraction (r= .85) methods.

Results indicated that the extraction method was faster, easier to perform and accurate as the distillation

method for monitoring lipid oxidation in poultry.

B) Nuggets

Sarkar et al. (2019) undertook a study to standardize processing protocol of traditional bhujia by

incorporation with spent hen meat powder and to evaluate the economics of developed product. Four

treatment combinations i.e. Treatment A (0% meat powder), Treatment B (10% meat powder),

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Treatment C (15% meat powder) and Treatment D (20% meat powder) were prepared to evaluate

economics of the bhujias. To analyse the economics of production, the total production cost of

treatments (per 100kg) were calculated on the basis of overhead production cost and formulation cost of

bhujia mix powder which was found to be Rs. 17,595.00, Rs. 26,085.00, Rs. 30,330.00 and Rs.

34,575.00 for Treatment A, B, C and D, respectively. The cost of formulation for 100 kg bhujia mix

powder was found highest for the Treatment D followed by Treatment C, B and A. Per day expenditure

for processing of 100 kg bhujia mix powder was found to be highest in Treatment D followed by

Treatment C, Treatment B and Treatment A. Maximum total net profit was found for Treatment D

followed by Treatment C, Treatment B and Treatment A. Break-even point were estimated as

Rs.1,95,006.00, Rs. 1,94,993.00, Rs. 1,95,012.00 and Rs.1,95,002.00 for Treatment A, B, C and D

respectively. The cost benefit ratio was estimated around 30% for all the treatments. Estimating the

details of economics of the developed products, it can be concluded that a viable enterprise can be

established by keeping MRP per kg of bhujia as Rs. 190, Rs. 261, Rs. 283 and Rs. 304 for Treatment A,

Treatment B, Treatment C and Treatment D respectively.

Shinde et al. (2019) prepared Japanese quail meat nuggets with addition of 6% Finger millet flour

(Eleusine coracana) and were compared with control. Cooked Japanese quail meat nuggets were packed

in low density polyethylene (LDPE) pouches and stored at refrigeration temperature (4±1°C).

Assessment for various quality parameters such as physico-chemical, proximate, microbiological and

sensory characteristics were conducted. Moisture, protein and fat content declined significantly with the

advancement of storage period, however, ash content increased significantly. The microbiological

parameters such as total plate count and yeast and mould count increased with the increasing days of

storage but were within the permissible limits. The psychrophilic count, however, was not observed up

to 20 days of storage. The sensory scores for all sensory characteristics declined progressively with the

advancement of storage period of 20 days. However, both control and 6% finger millet flour added

Japanese quail meat nuggets were acceptable up to 20th day of refrigerated storage.

Sujarwanta et al. (2019) conducted an experiment to find out the effect of curcuma flour fortification of

chicken nugget. Five formulations were developed with inclusion of curcuma flour to the chicken

nuggets at 0, 0.5, 1, 1.5, and 2% of filler (w/w). The chicken nuggets were analysed for their physical,

chemical and sensory characteristics. Curcuma flour fortification did not affect the water holding

capacity, tenderness, protein and fat contents of chicken nugget (P>0.05), but increased the vitamin E

and curcumin contents of chicken nugget (P<0.05). Sensory characteristic test results showed that the

24

fortification of curcuma flour did not affect acceptability in the sensory attributes of chicken nugget. In

conclusion, chicken nugget with curcuma flour fortification at the level of 2.0% was reported as the best

formulation for enhancing nutritional properties of the chicken nugget.

Anandh and Villi (2018) prepared restructured spent hen meat nuggets by using spent hen meat

emulsion and ground spent hen meat batter and their quality and sensory acceptability were evaluated.

Restructured emulsion nuggets had significantly (p<0.01) higher pH, emulsion stability, product yield

and drip loss while restructured spent hen meat nuggets prepared from ground spent hen meat batter had

significantly (p<0.01) higher moisture contents. Protein content between restructured emulsion and

ground spent hen meat batter nuggets did not differ significantly. Sensory evaluation scores for

appearance and colour, flavour, juiciness, tenderness, binding and overall acceptability (p<0.01) were

higher for restructured spent hen meat nuggets prepared from emulsion than spent hen meat nuggets

prepared from ground spent hen meat batter. The restructured spent hen meat nuggets prepared by

emulsion was rated from moderately to highly acceptable, whereas the restructured spent hen meat

nuggets prepared by ground spent hen meat batter was rated as moderately acceptable. In conclusion to

the study, spent hen meat could be successfully used for value addition in the preparation of chicken

nuggets.

Rindhe et al. (2018) evaluated the effect of hydrated wheat bran on physico-chemical, textural, color

and sensory qualities of spent hen meat nuggets. Moisture, protein, fat and aw decreased significantly

(P<0.05), while ash, fibre, water holding capacity, emulsion stability and cooking yield increased

significantly (P<0.05) with the incorporation of wheat bran. It was noted that addition of hydrated wheat

bran significantly (P<0.05) increased moisture retention and fat retention. The colour profile recorded

an increase in lightness (L*) and yellowness (b*) while redness (a*) decreased (P<0.05) with the

addition of hydrated wheat bran. The effect of addition of hydrated wheat bran to nuggets was also

observed in regard to textural properties. Hardness, chewiness and gumminess of spent hen meat

nuggets were higher, whereas springiness, stringiness, cohesiveness and resilience were lower in wheat

bran added nuggets than the control group. All the sensory properties decreased significantly (P<0.05)

with the addition of hydrated wheat bran. However, sensory panellists rated the nuggets on the basis of

quality as ‘very good to excellent’. The study successfully developed a fibre enriched spent hen meat

nuggets with the incorporation of pre-hydrated 8% wheat bran.

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Saini et al. (2018) undertook a study to standardize processing protocol of fried snack and to evaluate

economics of the developed product. Three treatments were prepared with incorporation of rosemary

extract (T1), betel extract (T2) and their 1:1 combination (T3) in the product by replacing chicken

powder (3%) to evaluate economics of the fried snack. All treatments and control group were deep fried

at 190°C for 45 sec. In estimating the cost of production, cost of formulation was found highest for

group T2. Maximum total net profit was found for control followed by T2, T4 and T1. The break-even

point was estimated at₹ 14,31,707.37 for control and ₹ 24,96,454.21, ₹ 16,33,154.67 and 21,88,047.71

for T1, T2 and T3, respectively. The cost benefits ratio was found highest for control and T3 whereas

T1 recorded the lowest. From the estimated economics of the developed product, the study concluded

that a viable enterprise could be established by keeping rate ₹ 261, ₹ 241, ₹ 260 and ₹ 230 for rosemary

extract, betel extract, their 1:1 combination incorporated product and control, respectively.

Sravadnya et al. (2018) conducted an economic study of value added Hot Extruded puffed product by

incorporation of spent hen meat emulsion in the flour in various proportions. Spent hen meat emulsion

and two flour mixtures (Corn flour and Rice flour) were admixed in 15:85 (T1), 20:80 (T2) and 25:75

(T3) proportions to obtained emulsion based dough to produce hot extruded puffed product by

effectively using twin screw hot extruder. Hot extruded puffed products thus obtained were rich in

protein because of incorporation of spent hen meat which constituted around 10-15% protein. Economic

survey was conducted by comparing the spent hen meat incorporated puffed product with other meat

products in the meat industry and adoption of most feasible methodology for production of cheapest

form of highly value added puffed products in meat industry.

Verma et al. (2018) in their study evaluated the effect of natural tenderizer on histological, physico-

chemical and sensory properties of raw and cooked emu meat. Fresh emu meats were procured from

local market and cut into small chunk of approximately 3cm3 size and were marinated with different

concentrations (0, 5, 7 and 10% w/v) of cucumis and papaya juice for 24 h at 4±1°C. Based on sensory

evaluation results, 10% w/v cucumis and papaya juice samples being noted as most acceptable were

allotted for further studies. The results of various studies showed remarkable variations between the

treated emu meat with plant proteolytic enzymes and the corresponding control. Sensory scores revealed

a significant improvement in flavor, tenderness and juiciness of treated samples compared to control

samples. It was also observed that the muscle fibre diameter and sarcomere length value were slightly

lower in the treated samples than control. Significant (P<0.01) reduction was noted in cooking yield and

pH of samples treated with cucumis in comparison to papaya. The shear force value was significantly

26

reduced (P<0.01) in all the treated samples. There was significant (P<0.01) improvement in flavor,

juiciness, tenderness and overall acceptability scores of all treated samples in comparison to control.

Arshad et al. (2017) aimed to investigate the effect of wheat germ oil and α-lipoic acid on the quality

characteristics, antioxidant status, fatty acid profile, and sensory attributes of chicken nuggets. Six types

of diets were prepared for feeding the chickens to evaluate the quality of nuggets made from the leg

meat of these experimental animals. These included control, diet enriched with wheat germ oil (WGO),

which is a rich natural source of α-tocopherol (AT), diet with added AT or α-lipoic acid (ALA), diet

with a combination of either ALA and WGO (ALA + WGO) or ALA and synthetic AT (ALA + AT).

ALA has great synergism with synthetic as well as natural AT (WGO). The diet with WGO and ALA

showed the best potential with respect to both antioxidant activity and total phenolic content. HPLC

results revealed that the chicken nuggets made from WGO + ALA group showed maximum deposition

of AT and ALA. The stability of the nuggets from control group was found to be significantly lower

than that of nuggets from the WGO + ALA group. Total fatty acid content too was higher in the nuggets

from this group. The poly unsaturated fatty acids (PUFA) were found to be higher in the nuggets from

the groups fed with a combination of natural and synthetic antioxidants. It was concluded that the

combination of natural and synthetic antioxidants in the animal feed exerts a synergistic effect in

enhancing the stability and quality of chicken nuggets.

Pathera et al. (2017a) conducted a study on the effect of dietary fibre enrichment (wheat bran) and

cooking methods (oven, steam and microwave) on the functional and physico-chemical properties of

raw nuggets formulation as well as nutritional, colour and textural properties of chicken nuggets.

Among the various cooking methods used for nuggets preparation, steam cooked nuggets had

significantly (p<0.05) higher water holding capacity (56.65%), cooking yield (97.16%) and total dietary

fibre content (4.32%) in comparison to oven and microwave cooked nuggets. The effect of cooking

methods and wheat bran incorporation were also noticed on textural properties of the nuggets. Hardness,

firmness and toughness values of oven and steam cooked nuggets were significantly (p<0.05) higher

than microwave cooked nuggets. Among nuggets prepared by different cooking methods, cohesiveness

of microwave cooked nuggets was found to be highest (p<0.05), whereas, oven cooked nuggets had the

highest gumminess and chewiness values (p<0.05). Steam cooked nuggets were found to be better

among all nuggets due to their higher cooking yield and dietary fibre content.

27

Akesowan (2016) formulated a low fat, high dietary fibre and phosphate free chicken nuggets by adding

various levels of a konjac flour/xanthan gum (KF/XG) (3:1) mixture (0.2–1.5 %, w/w) and shiitake

powder (SP) (1–4 %, w/w). A central composite rotatable design was used to investigate the influence

of variables on the physical and sensory properties of nuggets and to optimize the formulated nugget

formulation. The addition of the KF/XG mixture and SP was effective in improving nugget firmness and

increasing hedonic scores for colour, taste, flavour and overall acceptability. It was observed that the

nuggets became darker on addition with more SP. Optimal nuggets with 0.39 % KF/XG mixture and

1.84 % SP recorded reduction in fat, higher dietary fibre and amino acids. After storage at frozen

condition (-18±2°C), optimal formulated nuggets showed slower reduction in moisture, hardness and

chewiness in comparison to standard nuggets. Konjac flour and SP also recorded lowered lipid

oxidation in frozen formulated nuggets. Slight changes in sensory score were observed in both nuggets

and were found to be microbiologically safe after 75 days of frozen storage period.

Kumar et al. (2016) conducted a comparative study on the quality of nuggets prepared from different

combination of spent duck and spent hen meats in terms of physico-chemical and sensory attributes.

Nuggets from 75% spent duck and 25%spent hen (T1), 50% spent duck and 50% spent hen (T2) and

25% spent duck and 75% spent hen (T3) meats were prepared by standard methods. The quality

parameters study included pH, thiobarbituric acid (TBA), tyrosine value (TV), moisture (%), protein

(%), fat (%) and sensory attributes. Results revealed that pH, TV, and protein showed insignificant

difference between the treatments whereas TBA, moisture and fat varies significantly. Among the

sensory attributes colour, flavour, juiciness, texture, tenderness and overall palatability showed

significantly higher scores except appearance for nuggets made from T3 combination of meat. From this

study, it was concluded that the overall quality of nuggets prepared from higher ratios of spent hen meat

was better followed by other combinations of meat nuggets.

Pathera et al. (2016) conducted a study to evaluate the effect of oven, steam and microwave cooking

methods on lipid oxidation, microbiological and sensory quality of chicken nuggets. Nuggets were

prepared and analyzed at a regular interval of 5 days from the day of production to spoilage of products

under refrigerated storage. Cooking methods significantly affected the lipid oxidation of nuggets.

Highest lipid oxidation was reported in microwave cooked nuggets; however cooking methods did not

affect the microbiological quality of nuggets during storage. Products were safe for consumption up to

15 days at refrigerated storage as the microbial count was within the permissible limit. Steam cooked

nuggets showed better sensory scores in comparison to microwave and oven. Overall acceptability

28

scores for all the products were more than 6.0 at the end of storage period, reflecting more than

moderate acceptance till the products were microbiologically safe.

Reddy et al. (2016) carried out an experiment for comparing the meat quality attributes of spent breeder,

layers and broilers meat to assess the suitability for preparing the processed chicken meat products.

Spent breeder hen meat recorded significantly (P<0.05) lower drip loss, highest water-holding capacity,

lower shear force value, highest total protein extractability, lowest collagen, highest collagen solubility,

lower cooking loss, highest protein content and superior sensory scores in comparison to spent layer and

broiler meat. From the results obtained from the study, it was concluded that spent breeder meat was

superior to the meat of spent layers and broilers birds and more suitable for preparation of processed and

value added chicken meat product.

Cagdas and Kumcuoglu (2015) investigated the inhibitory effect of grape seed powder (GSP) on lipid

oxidation in chicken nuggets during frozen storage for 5 months. Chicken nuggets were prepared by

dipping into a batter containing GSP and pre-fried at 180°C and then stored at −18°C. Thiobarbituric

acid reactive substance (TBARS) values slowly increased during the first 2 months of storage and then

slightly decreased. However, at the end of the storage period, the levels were increased to 0.4

mgMDA/kg meat and were lowest in 10 % GSP (0.104 mg MDA/kg meat). Generally, samples treated

with GSP had lower POV, pAV, TBARS, and CD values in comparison to the control. These findings

indicated that GSP significantly (p<0.05) retarded lipid oxidation in pre-cooked chicken nuggets.

Parveez et al. (2015) prepared chicken nugget with chicken breasts, battered and deep-fried in

vegetable oil. The present study was undertaken to evaluate physico-chemical properties, viz., pH,

cooling yield (%), moisture (%), crude protein (%), ether extract (%), ash (%), crude fibre (%),

moisture- protein ratio, and coating thickness (cm), and sensory attributes, viz., colour &

appearance, flavour, juiciness, texture, and overall acceptability of chicken nuggets enrobed with

pea (Pisum sativum) flour at two different concentrations in the batter mix, viz., 25% w/w (Batter

mix-I) and 5% w/w (Batter mix-II), fortified with 6% papaya (Carica papaya ) pulp as the source

of fibre in the product. The study revealed that crude protein, ether extract, crude fibre, and coating

thickness were significantly (P<0.05) higher in the product enrobed with batter mix-II, as compared

to the product enrobed with batter mix-I. Conversely, moisture content and moisture-protein ratio

were higher in the product enrobed with batter mix-I, as compared to the product enrobed with

batter mix-II. The concentration of pea flour in the enrobing material had no effect (P>0.05) on

29

ash, pH, and cooling yield. The texture and overall acceptability score of the products enrobed

with batter mix-I were significantly (P<0.05) higher than the products enrobed with batter mix-

II. The study concluded that chicken nuggets enrobed with batter mix-I had higher consumer

acceptability than the chicken nuggets enrobed with batter mix- II and conversely, chicken nuggets

enrobed with batter mix-II showed higher nutritional values than the chicken nuggets enrobed

with batter mix-I.

Polizer et al. (2015) aimed to develop and evaluate a chicken nugget formulation with partial

substitution of the meat or fat by pea fibre. Three formulations were developed: Control (C) –

commercial formulation, Fibre Less Meat (FLM) – reduction of 10% of meat and addition of 2% of pea

fibre and Fibre Less Fat (FLF) – reduction of 10% of fat and addition of 2% pea fibre. The products

were characterized for their pH value, instrumental colour, texture, cooking loss (frying), proximate

composition and sensory properties (acceptance test). The control treatment presented lower (p<0.05)

pH values when compared to FLM and FLF. The analysis of cooking loss showed no differences

(p>0.05) amongst the treatments. The texture analysis showed no significant differences amongst the

treatments for elasticity and cohesiveness, although the FLF batch was firmer than the others (p<0.05).

Regarding the sensory acceptance test, the consumers observed no difference (p>0.05) amongst the

three treatments for aroma, texture, flavour or overall acceptability. From the study, it was observed that

it was possible to partially replace meat and fat by pea fibre in chicken nuggets, without compromising

most of the physicochemical characteristics or altering the sensory acceptance.

Verma et al. (2015) in their study evaluated the effect of salt substitution (Treat- I) and added pea hull

flour (PHF) at 8 (Treat-II), 10 (Treat-III) and 12 % (Treat- IV) levels on the quality of low fat chicken

nuggets (Control). Replacement of NaCl significantly affected (P<0.05) emulsion and product pH,

emulsion stability, cooking yield, ash content while PHF had additional effect on moisture and protein.

Dietary fibre content in the product significantly (P <0.05) increased at each level of PHF. The colour

parameters remained similar to control due to salt replacement while added PHF decreased their values.

Textural properties were lower (P <0.05) in the treated products. Addition of PHF significantly (P

<0.05) decreased cholesterol and glycolipids contents at 8 % and 12 % levels, respectively. Sensory

evaluation exhibited that 40 % NaCl can efficiently be replaced and 8 % PHF can be incorporated as a

source of fibre in low fat chicken nuggets without significant effect on various attributes.

30

Das et al. (2013) carried out a trial to prepare nuggets from the relatively tough and fibrous meat of desi

spent hen using fermented bamboo shoot as a phytopreservative in order to enhance the physico-

chemical, microbiological and keeping quality of the nuggets. Lean meat of desi spent hen was minced

and blended along with other non-meat ingredients and fermented bamboo shoot @10%. The emulsion

was filled in metallic moulds and steam cooked and cut into pieces. Ready-to eat nuggets thus prepared

were packed in sterilized LDPE zip bags and stored at 4±1°C up to 15 days for quality evaluation.

Emulsion stability (%), cooking yield (%) and proximate composition were studied on the day of

preparation, while estimation of pH, TBA values, microbial load and sensory evaluation were carried

out at 5 days interval and up to 15th day of storage. The emulsion stability (%), cooking yield (%),

moisture (%), crude protein (%) and total ash (%) of FBS treated nuggets differed significantly (p<0.01)

from the control products. Storage studies revealed significantly lower (p<0.01) pH, TBA value, total

plate count, psychrophillic count and counts for yeast and moulds in FBS treated nuggets in comparison

to control products. Both control and treated nuggets exhibited gradual loss of panel ratings during the

storage period (4±1°C for 15 days), however, nuggets containing fermented bamboo shoot revealed

significantly higher (p<0.01) mean sensory scores in terms of flavour, texture, juiciness and overall

acceptability. Nuggets with better physico-chemical and shelf life can be prepared with incorporation of

fermented bamboo shoot @10% (w/w) to the nugget emulsion.

Suradkar et al. (2013) studied to assess the comparative quality of chicken nuggets prepared from

broiler, spent hen and combination (50:50) meats in terms of physico-chemical and sensory attributes.

Chicken nuggets from broiler (T1), spent hen (T2) and combination 50:50 (T3) meats were prepared by

standard methods. The quality parameters studied included pH, moisture (%), protein (%), fat (%),

emulsion stability, cooking yield (%) (both raw and cooked product) and sensory attributes. Results

revealed that moisture, fat and cooking yield showed significant difference between the treatments.

Among the sensory attributes, juiciness, texture and overall palatability showed significantly higher

scores for nuggets made from broiler meat. Other attributes like the appearance and flavour in case of

T1 also showed higher score than the other treatments but there was no significant difference. From the

study, it was concluded that the overall quality of nuggets prepared from broiler meat was better

followed by combination meats and spent hen nuggets.

Kumar and Tanwar (2011) in an experiment prepared chicken nuggets from spent hen meat using

ground mustard as phyto-preservative without impairing the sensory attributes of the product. The

antioxidant and antimicrobial efficacy of mustard on keeping quality of the product was also assessed.

31

The emulsion stability (%), cooking yield (%) and moisture content (%) of the product containing

ground mustard differed significantly (p≤0.05) from the control. Nuggets containing ground mustard

maintained significantly (p≤0.05) higher sensory scores throughout the storage period (at 4±1 °C for 15

days). The pH as well as thiobarbituric acid value increased significantly (p≤0.05) with advancement of

storage period. Ground mustard maintained significantly lower thiobarbituric acid values throughout the

observation period than the control. Microbiological studies revealed significant increase in total plate

count and lipolytic count with the length of storage period. Microbial counts were found to be

significantly (p≤0.05) higher in control than in nuggets containing ground mustard.

Souza et al. (2011) aimed to evaluate the viability of the use of spent laying hens’ meat in the

manufacturing of mortadella-type sausages with healthy appeal by using vegetable oil instead of animal

fat. 120 Hy-line® layer hens were distributed in a completely randomized design into two treatments of

six replicates with ten birds each. The treatments were birds from light Hy-line® W36 and semi-heavy

Hy-line® Brown lines. Cold carcass, wing, breast and leg fillets yields were determined. Dry matter,

protein, and lipid contents were determined in breast and leg fillets. The breast and leg fillets of three

replicates per treatment were used to manufacture mortadella. After processing, sausages were evaluated

for proximal composition, objective colour, microbiological parameters, fatty acid profile and sensory

acceptance. The meat of light and semi-heavy spent hens presented good yield and composition,

allowing it to be used as raw material for the manufacture of processed products. Mortadellas were safe

from microbiological point of view, and those made with semi-heavy hens fillets were redder and better

accepted by consumers. Values for all sensory attributes were evaluated over score 5 (neither liked nor

disliked). Both products presented high polyunsaturated fatty acid contents and good polyunsaturated to

saturated fatty acid ratio. The excellent potential for the use of meat from spent layer hens of both

varieties in the manufacturing of healthier mortadella-type sausage was demonstrated.

Warhadpande et al. (2010) assessed the level of inclusion of chicken blood plasma (CBP) in the

preparation of cakes in respect of certain physico-chemical quality traits. The cakes were prepared with

and without added flavour. In each group, seven cakes were prepared from 0 (control) to 60% level of

inclusion of CBP with 10% interval. The cakes at 40% level of incorporation of CBP recorded highest

visual grades for colour and consistency. There was gradual rise in cake volume up to 40% level and on

further increase in level of inclusion of CBP resulted into subsequent fall in cake volumes. The pH of

cakes did not differ significantly up to 20% level but it increased beyond 20%. The moisture, total ash

and crude protein contents of cakes exhibited an increasing trend from 0 to 60% level of inclusion of

32

CBP. The ether extract of cakes showed a gradual decrease at increased level of inclusion of CBP.

There was no significant effect of flavour for all the parameters studied. Based on the overall results, it

may be concluded that CBP could be successfully used up to 30% level of inclusion for value addition

in egg products.

Contreras-Castilloa et. al. (2008) carried out a trial to characterise and compare the quality of the

mechanically separated meat (MSMs) obtained from carcasses from breeding and laying hens to provide

useful information for the processing industry. The composition of the mechanically separated meat

(MSM) varied between the two groups. Laying hens contained the most crude protein (P<0.05) and

high ash content (P<0.05). Calcium concentrations and bone contents were higher (P<0.05) for laying

(448 mg/100 g and 1.25%, respectively) than for breeding hens (299 mg/100 g and 0.78%, respectively).

Levels of unsaturated fatty acids for laying and breeding hens (75.89 and 72.82%, respectively) and

cholesterol concentrations (73 and 61 mg/100 g, respectively) were higher (P<0.05) for laying hens than

for breeding ones.

Reddy (2008) carried out a trial to study the effect of soy flour, corn flour and Bengal gram flour

separately each at 5, 10 and 15 % levels in the formulation of spent chicken meat nuggets. The study

revealed that 10% soy flour, 15% corn flour and 15 % Bengal gram flour formulations have lower

cooking losses, better emulsions stability, higher water holding capacity and superior sensory

characteristics compared to control and these were selected for pursuing the storage studies at

refrigerated storage (4±1°C). In the second phase, the selected flour formulations of spent chicken meat

nuggets were subjected to refrigerated (4±1°C) storage temperature and quality characteristics were

evaluated at every 4 days interval for a period of 20 days. The decreasing trend of water holding

capacity was noticed throughout the storage period. The mean ± S.E values of pH and 2-thiobarbituric

acid reactive substance values also significantly (P<0.05) increased with increase in refrigerated storage

were irrespective of flour formulations. The percent moisture and fat content of nuggets significantly

(P<0.05) decreased and percent protein content was significantly (P<0.05) increased during the storage

period. The percent total ash content was not affected significantly (P>0.05) by both flour formulations

and storage period. The total plate counts (log 10 CFU/ gm of meat sample) were significantly (P<0.05)

increased in the all flour formulations of spent chicken meat nuggets during the refrigerated (4±1°C)

storage period. The yeast and mould counts of nuggets significantly (P<0.05) increased in refrigerated

storage. Organoleptic evaluation scores revealed that the storage significantly (P<0.05) reduced scores

of all flour formulations of spent chicken meat nuggets. The cost of the production of nuggets extended

33

with 15% Bengal gram flour was lower followed by 15% corn flour followed by 10% soy flour and

control. The cost of the production (Rs/kg) of spent chicken meat nuggets prepared with control, 10%

soy flour, 15% corn flour and 15% Bengal gram flour were recorded at Rs. 150.00, Rs. 147.50, Rs.

134.00 and Rs. 132.00 respectively.

Biswas et al. (2006) conducted a study to compare and assess the quality of chicken and duck patties

prepared from broiler, spent hen and duck. The meat emulsions were analyzed for pH, moisture, protein,

fat, total plate count (TPC), total psychrophilic count (TPSC) and emulsion stability for raw emulsion

and in addition, the cooking yield, Thiobarbituric acid (TBA) value and sensory qualities such as,

appearance, flavour, tenderness, juiciness and overall acceptability were analysed for cooked patties.

The patties prepared from broiler meat showed significantly highest moisture content, emulsion stability

and cooking yield and on the other hand, the fat content was significantly highest in duck patties. The

TPC, TPSC, TBA values and sensory qualities of all the patties were within the acceptable level up to

14th day of refrigerated storage. There were no major drawbacks of the patties prepared from spent hen

and duck in comparison to those of broilers. From the study, the values of the major parameters were

observed to be within the range of standard values and in conclusion, the use of spent hen and duck meat

could be promoted for preparing nutritionally sound and acceptable patties.

Kumar and Sharma (2006) developed chicken patties from spent hen meat prepared from a standardized

formulation and were extended with barley flour (hydrated 1:l) at 0, 5, 10 and 15% levels replacing the

corresponding amount of lean meat. At 15% replacement level, pH, moisture, protein and fat percentage of

raw as well as cooked patties decreased as compared to 0% level (Pc0.05). A significant decrease (P<0.05)

in shear force value and per cent shrinkage of the product was recorded. The emulsion stability and cooking

yield improved significantly (P<0.05) with the level of extension with barley flour. The findings indicated

that chicken patties from spent hen meat could be extended with up to 10 per cent barley flour (hydrated 1:

l) in an economic formulation

34

CHAPTER III

OBJECTIVES & METHODOLOGY

Objectives:

i. To encourage layer poultry farmers by utilizing their spent hens so as to run the production

cycle smoothly.

ii. To develop technology for preparation of iron-enriched processed meat products from spent

hens, suiting to the convenience of school going children and women.

iii. To study the quality characteristics of newly developed spent chicken products.

iv. To propose an economic model for preparing iron-enriched products from spent hens for

small entrepreneurs.

v. Transfer of technology to the end-user.

Methodology:

Collection of spent hens: Healthy spent hens, quantum sufficit, were collected from layer egg

producing as per the availability after antemortem examination. These were brought to the

Laboratory, Dept. of Poultry Science, College of Veterinary Science, Khanapara for processing.

Processing of spent hens: The spent hens were scientifically slaughtered and processed. The carcass

was deboned to collect their meat, along with fat, preserved at -21°C until further use. The blood was

collected during the slaughter to which food grade anticoagulant Sodium citrate (citric acid) was

added @ 0.3g/100ml to prevent coagulation and was stored at refrigerated temperature (4±1°C) until

further use. The meat was minced using automatic mincer prior to the preparation of meat products.

Preparation of iron-enriched poultry meat products: Pilot studies were conducted for both

chicken meat balls and chicken nuggets, separately to evaluate the desirable and feasible levels of

blood incorporation to the final product. Instead of coagulated blood, whole blood was used to

ensure uniform mixing of the meat product with blood. The standardized basic recipe for the control,

respective to the treated chicken nuggets and chicken meat balls were maintained the same. For the

treated chicken nuggets, lean meat was replaced with whole blood at 11%, 14% and 17%. On the

other hand, for chicken meat balls, lean meat was replaced with whole blood at 5%, 7.5 and 10%.

The standardize formulation of chicken nuggets and chicken meat balls are given in the Tables below

(Table 1 and 2).

35

Table 1: Formulation for preparation of whole blood incorporated chicken nuggets

SL

NO

INGREDIENTS CONTROL

T1

(0% Blood)

TREATMENT

T2

(11% Blood)

TREATMENT

T3

(14% Blood)

TREATMENT

T4

(17% Blood)

1 Spent hen meat 71.5 60.5 57.5 54.5

2 Vegetable oil/

Poultry fat

5.0

5.0

5.0

5.0

3 Extender (Wheat

Flour)

5.0

5.0

5.0

5.0

4 Binder (Soya

Flour)

7.0

7.0

7.0

7.0

5 Spices 1.5 1.5 1.5 1.5

6 Condiments* 5.0 5.0 5.0 5.0

7 Blood 0 11.0 14 17.0

8 Ice flakes 5.0 5.0 5.0 5.0

9 Total 100 100 100 100

10 Salt 1.5 1.5 1.5 1.5

Table 2: Formulation for preparation of whole blood incorporated chicken meat balls

SL

NO

INGREDIENTS CONTROL

T1

(0% Blood)

TREATMENT

T2

(5% Blood)

TREATMENT

T3

(7.5% Blood)

TREATMENT

T4

(10% Blood)

1 Spent hen meat 73.5 68.5 66 63.5

2 Vegetable oil/

Poultry fat 5.0 5.0 5.0 5.0

3 Extender (Wheat

flour) 5.0 5.0 5.0

5.0

4 Binder (Soya

flour) 5.0 5.0 5.0

5.0

36

5 Spices 1.5 1.5 1.5 1.5

6 Condiments* 5.0 5.0 5.0 5.0

7 Blood 0 5.0 7.5 10

8 Ice flakes 5.0 5.0 5.0 5.0

9 Total 100 100 100 100

10 Salt 1.8 1.8 1.8 1.8

*Condiments used were garlic and ginger mixture in the ratio of 2:1.

Table 3: Formulation of Spice mixture

Sl. no Ingredients Parts

1 Anise 10

2 Black pepper 10

3 Cardamom 5

4 Cinnamon 4

5 Clove 1

6 Coriander 23

7 Cumin 23

8 Mace 2

9 Nutmeg 2

10 Red chilli powder 20

The minced chicken meat was mixed thoroughly with the non-meat ingredient according to the

formulation for both the chicken meat products and subsequently cooked. The prepared meat products

were then packaged under normal (aerobic) and vacuum condition and stored under refrigeration

temperature.

The standard procedures followed to prepare these poultry products are presented in Fig. 1 and Fig.2.

37

Slaughtering, processing and deboning of the chicken

Meat mincing

Grinding the meat with ingredients

Shaping into ball of 10g, each

Boiling or steam cooking (75-85° for 45mts)

Ready-to- eat chicken meat balls

Slaughtering, processing and deboning of the chicken

Meat mincing

Grinding the meat with ingredients

Fig. 1. Chicken Meat Balls

38

Filling tightly in aluminium or steel moulds with lids

Cooking in water (80°C for 45mts; 5lb pressure)

Cold showering to cool

Chilling (4°C)

Cutting (4cmX1.5cmX1.5cm)

Ready-to-eat Nuggets

1. Packaging, Quality evaluation of prepared products and Storage stability studies:

The freshly prepared chicken meat balls and chicken nuggets were divided into two lots, each.

The first lot was subjected to organoleptic evaluation by the panel of expert judges and zero day

analysis for iron and cholesterol content, physico-chemical, proximate and microbial status.

Second lot was packaged in UV sterilized food grade LDPE pouch (3 micron thickness;

0.46g/100 sq inch water vapour transmission rate) under two modules: (i) Normal/Aerobic and

(ii) Vacuum and were stored at two different temperature i.e., at room temperature and

refrigeration temperature (4±1°C) to monitor the physico-chemical and microbial changes during

storage at the interval of 5 days up to 15 days. The third packaging module i.e., Modified

atmosphere packaging (MAP) could not be carried out as proposed as the equipment went out of

order during the research period and unavailability of other facility in the institute compelled this

portion of work to be discontinued.

Fig. 2. Chicken Nuggets

39

2. Organoleptic evaluation: Ready-to-eat chicken meat balls and chicken nuggets incorporated

with different levels of whole blood were subjected to evaluation for organoleptic qualities by

serving them to a semi-trained panel of 9 members. Panel members were allowed to sit in a well-

lighted and ventilated room. A clean glass of drinking water was offered to each member for

rinsing the mouth before and after testing of each product. They were neither informed about the

identity of the products nor allowed any conversation or discussion among themselves during the

evaluation process. All the samples were evaluated for colour, flavour, texture, juiciness and

overall acceptability by using a 7-point hedonic scale (Annexure-I) as described by Bratzler,

1971.

3. Estimation of iron content: The iron content in the meat product was estimated by following the

method of Narain & Ilango (2015) with slight modification which is based on thiocyanate

spectrophotometric technique.

4. Estimation of cholesterol content: The cholesterol content of the meat products was determined

by following the method as described by Rajkumar et.al. (2004).

5. Proximate composition of the poultry meat products: The proximate compositions of the

sausages were determined on as such basis as per methods of AOAC (2007) for a) Moisture b)

Crude Protein and c) Ether Extract (Fat)

6. Physico-chemical characteristics:

a) pH: The hydrogen ion concentration (pH) of the poultry meat products were determined by

following the method as described by Pippen et al. (1965)

b) Water holding capacity (WHC): Water holding capacity was determined using centrifuged

technique (Wardlaw et.al., 1973).

c) Thiobarbituric acid (TBA) value: The TBA values of the chicken meat balls and nuggets

were determined at different periods of storage by the method as described by Witte et al.

(1970).

d) Tyrosine value (TV): In the presence of tyrosine, folin ciocalteu phenol reagent produces a

blue color, the intensity of which is a measure of protein cleavage (Strange et al., 1977)

7. Microbiological Studies: Total viable plate count, yeasts and moulds count and incidence of

Salmonellae of samples at all stages of storage up to 15 days were estimated following the

standard method of APHA (1992).

8. Development of a small-scale poultry product preparation model unit:

A ‘small-scale poultry product preparation model’ unit was developed so as to encourage

educated unemployed youths to attract towards starting entrepreneurship for the preparation of

40

iron-enriched poultry products, i.e., meat balls and meat nuggets commercially.

An outline of ‘small-scale poultry product preparation model’ is given herewith, under

Annexure- II.

9. Entrepreneurship Training programme:

Two entrepreneurial training programmes were organized at the Department of Poultry Science,

College of Veterinary Science, Khanapara, Guwahati on Feb. 27/2020 and March 07/2020. For

this, educated unemployed youths having aptitude towards starting of entrepreneurship were

selected from different districts of the state, randomly. A total of 18 numbers of participants

attended the two training programmes. On submission of the project proposal, it was envisioned

to impart training programmes to three groups. However, due to prevailing Lockdown condition

in the context of Covid-19 only two target groups could be managed. They were given three-day

long trainings on the preparation of iron-enriched poultry products so as to start small-scale

production unit as business enterprise.

10. Statistical analysis: The data on all parameters were analyzed using appropriate statistical

analytical methods to arrive at a valid conclusion.

11. Deliverables:

It becomes evident from the study that the meats from spent hen could be used to prepare

iron enriched meatballs and nuggets employing simple technology. Thus, it will address the

imminent problem of the upcoming layer farmers so as to dispose off their spent layers stock

after completion of each laying cycle.

It may boost the entrepreneurship promotion among the unemployed youths of this region in

the context of novel meat product production and marketing.

It may boost the nutritional security of children and women.

Consumption of value added meat products will improve the overall health of human

resources in the society.

41

CHAPTER IV

RESULTS & DISCUSSION

Pilot Study:

Series of pilot studies were conducted to ascertain the level of blood to be incorporated in either

of the products, chicken meatballs and chicken nuggets. This was done to incorporate the

maximum level of iron, in the form of blood in the product, however without compromising the

taste and consistency of the products. For this, semi-trained panelists were selected and above

products were offered and allowed to judge on the basis of 7-point Hedonic scale. The taste

panelists opined that a maximum of 10.0 % blood can be incorporated in chicken meat balls.

Beyond this level the ball could not maintain desired ball shape. This may be due to the

loosening the binding ability of meat-dough for higher incorporation of blood as the elevated

level influenced on moisture content of the product and interfered in binding properties. On the

other hand, in case of chicken nuggets the level could be taken up to 17 % level. Since the

product was prepared in metallic mould the binding ability of meat-dough had little impact.

However, when the level of blood was used beyond 17 per cent level, the panelist experienced

metallic flavour and the product received lower than 5 (good) score.

After the pilot study, chicken nuggets were prepared with whole blood incorporated at

11%, 14% and 17% and chicken meatballs were prepared with whole blood at 5%, 7.5 and 10%

as per the standardized formulation and standard procedure, The freshly prepared chicken

meatballs and chicken nuggets, divided into two lot each where first lot were subjected to

organoleptic evaluation by the panel of expert judges and zero day analysis for iron and

cholesterol content, physico-chemical, proximate and microbial status. Second lot were packaged

in UV sterilized food grade LDPE pouch under two modules: (i) Normal/Aerobic and (ii)

Vacuum and were stored at room and refrigeration temperature (4±1°C) and the storage stability

studies were monitored at an interval of 5 days up to 15 days. The results and observations are

given in details below.

Iron content

The estimation of iron content for chicken meatballs and chicken nuggets were conducted and

the results are shown in the table 1.01 and 1.02, respectively.

42

Table 1.01: Iron content (mg/100mg) of chicken meatballs incorporated with whole blood

(Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

36.40a±0.49 63.33 b±1.47 72.66 c±0.87 92.66 d±0.47

Means bearing different superscripts differ significantly (P<0.05)

In chicken meat balls, the group with the highest blood level (10%) showed a

significantly (P<0.05) high iron content (mg/100g) of 92.66±0.47 while the lowest was recorded

in control group at 36.44±0.49. The iron content significantly (P<0.05) increased in proportion to

the increasing level of blood inclusion. It was interesting to note that the 10% blood

incorporation in meat emulsion could increase the iron content to the extent of 154.56 per cent

when compared to the control group.

Table 1.02: Iron content (mg/100mg) of chicken nuggets incorporated with whole blood

(Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

37.07a ±0.45 105.46b ±0.38 119.20c±0.65 135.20d±2.1

Means bearing different superscripts differ significantly (P<0.05)

The results of iron estimation in the chicken nuggets showed that the group with highest

blood level (17%) recorded iron content at 135.20±2.1 mg/100g while the control group had the

lowest iron content of 37.33±0.45 mg/100g. A significantly (P<0.05) increasing trend of iron

content was also observed with the increase in the blood inclusion levels in the chicken nuggets.

A remarkable increased in the iron content was noted where an increase from 184.49 per cent

(11% blood) to 264.72 per cent (17%blood) was recorded on comparison to its control

counterparts. Thus, from the present study, it was evidenced that through the incorporation of

blood, the iron level could be elevated to an appreciable degree in both the meat products.

43

Blood, a good source of nutrients, has a high content of essential amino acids and high

bioavailability of iron namely heme iron and it is considered as non-allergenic protein when

compared to dairy and soy proteins ((Sorapukdee and Narunatsopanon, 2017). Heme iron is

better absorbed than nonheme iron (Walter et al., 1993; Kikafunda and Sserumaga, 2005).

Whole animal blood has traditionally been used in Europe and Asia in such products as blood

sausage, blood pudding, blood cake, and blood curd (Liu, 2002). Slaughterhouse blood is rich in

heme iron and utilization of more blood for human consumption should be of nutritional

advantage (Oellingrath and Slinde, 1985).

Moisture content

Proximate composition with respect to moisture in chicken meatballs and chicken nuggets are

shown in Table 2.01 and 2.02, respectively.

Table 2.01: Moisture content (%) of chicken meatballs incorporated with whole blood

(Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

63.52a ±0.04 64.00b±0.03 64.68c±0.04 65.24d±0.07

Means bearing different superscripts differ significantly (P<0.05)

Table 2.02: Moisture content (%) of chicken nuggets incorporated with whole blood

(Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

63.82a±0.08 65.34b±0.05 65.82c±0.07 66.08d±0.1

Means bearing different superscripts differ significantly (P<0.05)

From the results, it was very clear that the moisture percentage significantly (P<0.05)

increased in proportion to the addition of blood in both the meat products. This is because over

44

90 per cent of blood plasma is water, while less than 10 per cent is dissolved substances, mostly

proteins. A high water content of blood must have influence the end results. The findings of the

study are in agreement with Chowdhury et. al. (2015) who also reported of increasing moisture

percentage with increasing blood level in pork sausage incorporated with blood.

Fat content

The results of the proximate analysis with respect to fat % (as such basis) for chicken meatballs

and chicken nuggets are depicted in Table 3.01 and 3.02, respectively.

Table 3.01 Fat content (%) of chicken meatballs incorporated with whole blood (Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

9.08d±0.02 8.86c±0.01 8.02b±0.07 7.65a±0.03

Means bearing different superscripts differ significantly (P<0.05)

In chicken meat balls, it was observed that the fat content (%) significantly (P<0.05)

decreased in proportion to the increasing level of blood incorporation from 5 per cent to 10 per

cent. In the control group, the fat content was found to be 9.08±0.02 per cent; whereas, in the

group with highest level of blood (10%) the fat content was recorded at 7.65±0.03 per cent.

Table 3.02: Fat content (%) of chicken nuggets incorporated with whole blood (Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

8.96d±0.03

7.19c±0.04

6.78b±0.01

6.61a±0.03

Means bearing different superscripts differ significantly (P<0.05)

The fat content (%) of chicken nuggets in the control group recorded the highest (P<0.05)

value of 8.96±0.03; whereas, the lowest (P<0.05) was recorded in 17% blood incorporated group

45

(6.61±0.03). The values followed the same trend to that of meat balls. This may be due to the

fact that blood has a very low fat content and the replacement of lean meat with the increasing

level of blood may have influenced the results. Similar results were also reported by Chowdhury

et. al. (2015) and Hazarika and Biro (1993).

Crude protein content

The proximate composition (as such basis) of chicken meatballs and chicken nuggets

with regard to protein content are depicted in Table 4.01 and 4.02, respectively.

Table 4.01: Crude Protein content (%) of chicken meatballs incorporated with whole

blood (Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

19.34a±0.25 20.38b±0.03 21.65c±0.14 22.23d±0.09

Means bearing different superscripts differ significantly (P<0.05)

The crude protein of chicken meatballs showed a clear picture of enhancement in protein

content with the incorporation of blood which increased significantly (P<0.05) with the

increasing blood levels. The highest protein content (%) was recorded in T4 (10% blood) group

at 22.23±0.09. Apart from the contribution of protein from the chicken meat, the high protein

content of the blood must have influenced the results. It has been reported that chicken blood

contains about 87-88% crude protein on dry matter basis (Sorapukdee and Narunatsopanon,

2017; Liu, 2002) which depicts the high protein nature of blood. Blood is sometimes referred to

as liquid protein (Ockerman and Hansen, 2000).

Table 4.02: Crude Protein content (%) of chicken nuggets incorporated with whole blood

(Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

21.07a±0.10 22.56d±0.15 22.02c±0.10 21.85b±0.07

Means bearing different superscripts differ significantly (P<0.05)

46

In case of chicken nuggets, it was observed that all the blood incorporated groups showed

a significantly (P<0.05) higher protein content than its control counterpart. The increase in

protein content is attributable to the added blood which is naturally high in protein. However,

when compared within the blood incorporated groups, it was noted that with the increment in

levels of blood, there was a significant (P<0.05) fall in protein content (%) in T4 (21.68±0.07)

and T3 (22.02±0.10) groups when compared to T2 (22.56±0.15). This may be due to the

replacement of lean meat in proportion with the increasing blood level, thus, affecting the overall

protein content of the nuggets as lean meat constitutes the major portion of the nuggets. The

chicken broiler meat is also reported to contain 88.49% protein (Straková et al., 2011) on dry

matter basis.

Total cholesterol

The total cholesterol content of chicken meatballs and chicken nuggets are shown in Table 5.01

and 5.02, respectively.

Table 5.01: Total cholesterol content (mg/100 g) of chicken meatballs incorporated with

whole blood (Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

23.74d±0.38 21.11c±0.13 19.72b±0.28 18.29a±0.27

Means bearing different superscripts differ significantly (P<0.05)

In the chicken meatballs samples, there was a significant (P<0.05) decrease in the

cholesterol content (mg/100g) with the increasing level of blood levels. The highest value was

recorded for control group (23.74±0.38 mg/100g) and the least in T4 group (18.29±0.27

mg/100g).

47

Table 5.02: Total cholesterol content (mg/100 g) of chicken nuggets incorporated with whole

blood (Mean±SE)

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

22.18c±0.94 17.34b±0.15 15.98a±0.18 15.19a±0.16

Means bearing different superscripts differ significantly (P<0.05)

The results of cholesterol content (mg/100g) of chicken nuggets reflected a significantly

(P<0.05) lower cholesterol level in all the blood incorporated groups on comparison with the

control group. It was also noteworthy within the blood incorporated groups that T2 was

significantly higher than T3 and T4; whereas, there was no significant difference between T3 and

T4, though T4 was numerically lower.

An interesting observation was noted that in both the meat products, the level of

cholesterol decreased with the increase in blood level. The lowest fat % in both the meat

products were observed in the groups having the highest blood level might be due to the lower

fat content of the blood as discussed earlier. The cholesterol content of chicken meat (breast and

leg) ranges from 59 to 84 mg/100g (Dinh et al., 2011). Therefore, the increasing replacement of

lean meat in proportion to the blood inclusion further lowered the cholesterol content as well as

fat, thus having a direct affect on the overall level of cholesterol reflecting lower values.

Organoleptic quality

The organoleptic quality (colour, flavour, juiciness, texture and overall acceptance) scores of

chicken meatballs under aerobic and vacuum package conditions are given in the Tables 6.01.01

through 6.01.05.

48

Table 6.01.01: Colour scores (Mean±SE) of chicken meatballs at different blood levels

under aerobic or vacuum packaged condition (4±1°C)

Storage period (Days)

Packaging

condition

CONTROL T1

(0% blood)

TREATMENT T2

(5% blood)

TREATMENT T3

(7.5% blood)

TREATMENT T4

(10% blood)

0 Aerobic 6.00±0.31 6.57±0.20 6.71±0.18 5.71±0.42

5

Aerobic 6.00±0.31 6.50±0.24 6.64±0.18 5.71±0.42

Vacuum 6.00±0.31 6.50±0.18 6.64±0.18 5.71±0.42

10

Aerobic 5.93±0.23 6.14±0.46 6.50±0.24 5.64±0.32

Vacuum 6.00±0.22 6.36±0.28 6.64±0.18 5.71±0.43

15

Aerobic 5.57±0.38 6.00±0.24 6.29±0.29 5.43±0.23

Vacuum 5.64±0.18 6.21±0.34 6.50±0.15 5.57±0.35

The freshly prepared chicken meatballs exhibited a higher colour score in T2 and T3

groups in comparison with control group while T4 group scored the lowest. However, no

significant difference was observed between the groups. The higher scores observed in the T2

and T3 group maybe because of the darker colour in comparison to the pale colour of the control

group. However, the panelist rated the meatballs with 10% whole blood with a lower score of

5.71 which may be due to the unnaturally darker colour and less appealing appearance. The dark

colour is attributed to haemoglobin present in the blood which is one of the main colour

contributors to meat products (Oellingrath and Slinde, 1985) and on oxidation it turns dark

brown.

No significant difference was seen in colour score among the groups and the packaging

system during the storage period up to 15th day. However, the colour score gradually decreased

numerically in all the groups with increase in the storage time. It was also noted that the colour

of the meatballs packaged under vacuum system showed a better colour score in comparison to

aerobic packaged meat balls. An acceptable colour of blood-containing products is necessary

before it can be introduced to the consumers as blood products are dark in color, since blood

49

contains about 14% hemoglobin (Warris and Rhodes, 1977). The dark color is the main sensory

quality problem when blood is added to meat products (Mielnik and Slinde, 1983).

Table 6.01.02: Flavour scores (Mean±SE) of chicken meatballs at different blood levels

under aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic B7.00d±0.00 6.50ab±0.019 6.29ab±0.29 5.93a±0.23

5

Aerobic AB6.86b±0.09 6.50ab±0.15 6.21ab±0.15 5.71a±0.29

Vacuum B7.00b±0.00 6.50ab±0.19 6.29ab±0.29 5.79a±0.26

10

Aerobic AB6.50b±0.19 6.14ab±0.34 6.00ab±0.22 5.50a±0.29

Vacuum AB6.71b±0.15 6.29ab±0.21 6.14ab±0.14 5.57a±0.23

15

Aerobic A6.07a±0.25 5.93a±0.07 5.79a±0.21 5.29a±0.26

Vacuum AB6.71b±0.10 6.29ab±0.14 6.14ab±0.20 5.57a±0.28

Means bearing different superscripts column-wise (Lower case) or row-wise (Upper case) differ

significantly (P<0.05)

The freshly prepared chicken meatballs of the control group T1 exhibited significantly

(P<0.05) higher flavor score than all the blood incorporated groups (T2, T3 and T4) and T4

received the lowest scores of 5.93±0.23. The lower flavor score of blood incorporated groups

may be due to the characteristic taste of blood (Oellingrath and Slinde, 1985), making it less

palatable and with the increasing amount of whole blood influencing for undesirable flavor as

stated by Ockerman and Hansen (2000). With the advancement of the storage period i.e., up to

15 day, the gradual decrease in the flavor score was witnessed in all the four groups but the

significant difference (P<0.05) was observed only for control group on 15 day in the aerobic

packaged group. It is also noted that the vacuum packaged meatballs of all the groups showed a

50

better flavor score in comparison to the aerobic packaged groups throughout the storage period.

This may be due to lower lipid oxidation than its aerobic counterpart.

Table 6.01.03: Juiciness scores (Mean±SE) of chicken meatballs at different blood levels

under aerobic or vacuum packaged condition (4±1°C)

Storage

period (Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic 6.00±0.24 6.29±0.29 6.50±0.15 6.14±0.14

5

Aerobic 5.93±0.23 6.21±0.15 6.29±0.21 5.93±0.23

Vacuum 6.00±0.22 6.29±0.18 6.43±0.17 6.00±0.22

10

Aerobic 5.79±0.21 5.93±0.23 6.14±0.34 5.79±0.26

Vacuum 5.93±0.17 6.07±0.25 6.21±0.26 5.86±0.28

15

Aerobic 5.50±0.19 5.64±0.21 5.79±0.36 5.57±0.32

Vacuum 5.71±0.38 5.86±0.34 6.00±0.15 5.71±0.26

In regard to the juiciness of the freshly prepared meatballs, there was no significant

(P>0.05) difference among the groups; however, the panelists rated 6 to 7 scores (Very Good to

Excellent) in all the blood incorporated groups, T3 scoring the highest (6.50±0.15). On the other

hand, the control group was rated with a score of 6.00±0.24 which may be due to higher cooking

loss causing more sensation of dryness than its counterparts. The better scores in the blood

incorporated groups may be due to the higher moisture content in the product contributed by the

whole blood and lower cooking loss which might have influenced the juiciness. It is known that

blood comprises of 75-82% moisture (Leoci, 2014). On the contrary, the T4 group with 10%

whole blood received the lowest score (6.14±0.14) in comparison to T2 and T3. This may be due

to the sodden consistency of the product in proportion to high level of moisture making it less

palatable.

The juiciness score of the products gradually decreased with the extension of storage

period up to 15 days at 4±1°C regardless of the blood levels or packaging system, though not

significantly. This may be due to lower water holding capacity resulting from denaturation of

myofibrillar proteins and loosening up of micro structure of muscles allowing more water to be

51

entrained (Nagamallika et al., 2006). Nevertheless, the vacuum packaged groups consistently

gave better scores throughout the storage time than the aerobic counterpart.

Table 6.01.04: Texture scores (Mean±SE) of chicken meatballs at different blood levels

under aerobic or vacuum packaged condition (4±1°C)

Storage

period (Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic 7.00b±0.00 6.50ab±0.19 6.21ab±0.15 5.86a±0.28

5

Aerobic 6.93b±0.07 6.43ab±0.20 6.00a±0.22 5.71a±0.18

Vacuum 7.00b±0.00 6.50ab±0.19 6.14a±0.24 5.79a±0.21

10

Aerobic 6.71b±0.15 6.14ab±0.26 5.71a±0.21 5.50a±0.33

Vacuum 6.71c±0.18 6.43bc±0.17 5.86ab±0.34 5.57a±0.32

15

Aerobic 6.50b±0.15 5.93ab±0.20 5.64a±0.36 5.36a±0.18

Vacuum 6.64b±0.14 6.21ab±0.21 5.64a±0.18 5.43a±0.22

Means bearing different superscripts column-wise differ significantly (P<0.05)

The texture score of the meatballs was found to be significantly (P<0.05) high in control

group (7.00±0.00); whereas, in the blood incorporated groups the score decreased. This could be

due to the higher moisture content resulting in loose consistency and less chewy texture of the

products, thereby, affecting its palatability. On 5th day, the T3 and T4 groups received a

significantly (P<0.05) lower texture scores than the control group and similar trend was seen up

to 15 day of storage under both packaging systems. However, no significant difference was

observed between the control and T2 group (both aerobic and vacuum) throughout the storage

period. Among the blood inclusion groups, there was no significant difference in the texture

throughout the storage period regardless of packaged system, except on 10th day between

vacuum packaged T2 and T4 groups.

Notwithstanding the packaging system or the blood level, the fall in texture score was

observed to be directly proportionate with the gradual increase in the storage period up to 15

days at 4±1°C; however with no significant difference. Apparently, the lower texture score was

52

noticed with the increasing blood levels. Though the packaging system was not found to have a

pronounced affect on the discussed trait, yet the vacuum packaged meatballs showed a better

texture score as compared to the aerobic packaged counterparts.

Table 6.01.05: Overall acceptability scores (Mean±SE) of chicken meatballs at different

blood levels under aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic B7.00±0.00 B6.86±0.09 B6.64±0.18 B6.36±0.18

5

Aerobic B7.00b±0.00 AB6.64ab±0.18 AB6.50ab±0.29 AB6.00a±0.22

Vacuum B7.00b±0.00 B6.79ab±0.2 B6.64ab±0.21 B6.21a±0.26

10

Aerobic AB6.43±0.30 A6.14±0.24 AB6.29±0.10 AB5.71±0.21

Vacuum AB6.71±0.18 AB6.50±0.19 AB6.36±0.28 AB6.00±0.22

15

Aerobic A6.00±0.24 A6.00±0.22 A5.79±0.24 A5.43±0.28

Vacuum AB6.29±0.10 AB6.21±0.15 AB6.14±0.09 AB5.71±0.15

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

Panelists rated the overall acceptability of the freshly prepared meatballs with Excellent

score (7.00±0.00) for control group. Though all the treated groups (T2, T3 and T4) received

lower scores but their acceptability level remained comparable to that of control.

Till 5th day, the control group could be able to maintain its Excellent character; whereas

the score was significantly (P<0.05) lower among all the three treated groups under aerobic

condition. The control group could be able to retain its comparable quality till 10th day in aerobic

condition or till 15th day in vacuum condition. The overall acceptability score of the meat

products decreased gradually with storage which significantly (P<0.05) pronounced by 15th day

in all the groups under aerobic packaging system. Albeit decreasing overall acceptability score,

the meatballs packaged under vacuum system could score better in comparison to its aerobic

packaged counterpart.

53

The overall acceptability from the above results reveals that an addition of whole blood

up to 7.5% to chicken meatballs is possible as the score showed comparable results with that of

control group. It was also noted that the meatballs retained its desirable quality (‘Very good to

Excellent’) up to 10 days of storage. Additionally, vacuum system was observed to be a better

packaging option for maintaining the sensory qualities for a longer duration.

The outcome of using whole blood in meat processing is the final product with dark color

which may have negative effect on the appearance of the product. The amount of whole blood

used in meat and other food products tends to be relatively low as increasing the proportion has a

detrimental effect on sensory qualities, particularly flavor and color (Ockerman and Hansen,

2000). The traditional use of whole blood is generally restricted to products such as blood

sausages where the black color is both expected and acceptable (Ockerman and Hansen, 2000).

The organoleptic quality (colour, flavour, juiciness, texture and overall acceptance)

scores of chicken nuggets prepared from meat emulsions incorporated with whole blood at levels

from 11% up to 17% and preserved under aerobic or vacuum package conditions are given in the

Tables 6.02.01 through 6.02.05.

Table 6.02.01: Colour scores (Mean±SE) of chicken nuggets at different blood levels under

aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL

T1

(0% blood)

TREATMENT

T2

(11% blood)

TREATMENT

T3

(14% blood)

TREATMENT

T4

(17% blood)

0 Aerobic 7.00c±0.00 6.86bc±0.09 6.14ab±0.34 5.50a±0.27

5

Aerobic 7.00c±0.00 6.71bc±0.18 6.00ab±0.19 5.43a±0.30

Vacuum 7.00c±0.00 6.79bc±0.15 6.14ab±0.21 5.50a±0.39

10

Aerobic 6.79c±0.10 6.43bc±0.23 5.79ab±0.24 5.14a±0.18

Vacuum 6.86c±0.09 6.64bc±0.18 5.93ab±0.13 5.36a±0.21

15

Aerobic 6.71c±0.15 6.14bc±0.18 5.64ab±0.26 5.00a±0.19

Vacuum 6.79c±0.10 6.21bc±0.26 5.71ab±0.34 5.14a±0.09

Means bearing different superscripts column-wise differ significantly (P<0.05)

54

The panelists gave the highest colour scores (7.00±0.00) to the control group, which

remained comparable throughout the storage period of 15 days. The nuggets prepared by

incorporating 11% blood could be able to get significantly (P<0.05) similar score to that of

control group, throughout the said storage period. In comparison to the control group the colour

scores of T3 and T4 groups were significantly (P<0.05) lower. This indicates that the panelists

did not appreciate more intense colour of the nugget beyond 11%. It can be noted that the heme

component of the blood on oxidation imparts a characteristic undesirable brown colour in the

product.

All the groups under both aerobic and vacuum packaged systems showed non-significant

difference in the colour score with the extension of storage period up to 15 days stating no

impact of packaging system. However, beyond 10 days of storage showed significantly (P<0.05)

lower score.

Table 6.02.02: Flavour scores (Mean±SE) of chicken nuggets at different blood levels under

aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic 7.00b±0.00 6.36ab±0.18 6.21ab±0.26 5.43a±0.20

5

Aerobic 6.79b±0.15 6.36b±0.18 6.14ab±0.14 5.36a±0.18

Vacuum 6.86b±0.14 6.43b±0.23 6.21ab±0.26 5.36a±0.18

10 Aerobic 6.50b±0.27 5.93ab±0.28 5.71ab±0.34 5.14a±0.36

Vacuum 6.71b±0.15 6.14b±0.26 6.00ab±0.36 5.14a±0.28

15 Aerobic 6.21b±0.26 5.50ab±0.29 5.36ab±0.24 4.86a±0.24

Vacuum 6.50b±0.11 5.71ab±0.21 5.50a±0.29 4.93a±0.35

Means bearing different superscripts column-wise differ significantly (P<0.05)

Addition of whole blood in preparing chicken nuggets modified the flavor, especially

with the highest level of blood incorporation (17%). Since the heme component of the blood

imparts metallic taste and odor in the product, proportionately high level of blood might have

55

impacted on the flavor score. The nuggets containing 11% blood received a similar score to that

of control group throughout the storage period. On storage at 4±1°C up to 15 days, no significant

difference in the flavor was recorded in all the four groups regardless of the blood levels or

packaging methods.

Table 6.02.03: Juiciness scores (Mean±SE) of chicken nuggets at different blood levels

under aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL T1

(0% blood)

TREATMENT T2

(11% blood)

TREATMENT T3

(14% blood)

TREATMENT T4

(17% blood)

0 Aerobic B6.86±0.09 B7.00±0.00 B6.71±0.18 B6.50±0.19

5

Aerobic B6.86±0.09 B6.86±0.14 B6.64±0.18 B6.36±0.24

Vacuum B6.86±0.14 B6.86±0.14 B6.64±0.18 B6.43±0.20

10

Aerobic AB6.43±0.17 AB6.43±0.20 AB6.14±0.26 AB5.79±0.31

Vacuum AB6.50±0.19 AB6.50±0.22 AB6.21±0.26 AB5.86±0.34

15

Aerobic A5.71±0.34 A5.93±0.25 A5.71±0.26 A5.36±0.28

Vacuum A5.93±0.07 AB6.21±0.10 AB6.00±0.15 AB5.79±0.21

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

The juiciness score of the freshly prepared nuggets recorded a numerically higher score

in T2 (11% blood) group when compared to control group (7.00±0.00 vs 6.86±0.09) as depicted

in Table 6.02.03. This might be due to higher moisture content contributed by the blood and

lower cooking loss, thus making it juicier than the control. Conversely, more inclusion of blood

(14% and 17%) in the product could not attract more score which may be due to the sodden

consistency in proportion to high level of moisture, making it less palatable. However, no

significant difference was observed among the groups throughout the storage period, irrespective

of the packaging system. It was observed that the juiciness attribute of the products gradually

decreased with the increase in storage time up to 15 days at 4±1°C regardless of the blood levels

or packaging methods.

56

The juiciness score remained significantly (P<0.05) high till the 5th day of storage.

Thereafter, despite the diminished scores, it remained between Good to Very good, as judged by

the panelists. This may be attributed to decrease in the water holding capacity resulting from

denaturation of myofibrillar proteins and loosening up of micro structure of muscles allowing

more water to be entrained (Nagamallika et al., 2006). The influence of packaging methods on

the juiciness was insignificant; however, the panelists conferred higher score to the products

packaged under vacuum method as compared to its aerobic counterpart throughout the storage

period.

Table 6.02.04: Texture scores (Mean±SE) of chicken nuggets at different blood levels under

aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL T1

(0% blood)

TREATMENT T2

(11% blood)

TREATMENT T3

(14% blood)

TREATMENT T4

(17% blood)

0 Aerobic 7.00b±0.00 B6.79ab±0.15 C6.50ab±0.19 B6.07a±0.23

5

Aerobic 7.00b±0.00 B6.64ab±0.18 BC6.29ab±0.26 AB5.93a±0.28

Vacuum 7.00b±0.00 B6.71ab±0.15 BC6.43ab±0.20 AB5.93a±0.23

10

Aerobic 6.64b±0.18 AB6.14ab±0.14 ABC5.86ab±0.18 AB5.50a±0.33

Vacuum 6.71b±0.18 AB6.36ab±0.28 ABC5.86a±0.39 AB5.64a±0.32

15

Aerobic 6.36b±0.14 A5.71ab±0.18 A5.36a±0.28 A5.14a±0.09

Vacuum 6.57b±0.17 AB6.29ab±0.15 AB5.64a±0.28 AB5.43a±0.30

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

The control group could attract the maximum score of 7.00±0.00 on texture of the freshly

prepared nuggets (Table 6.02.04). However, as the blood level inclusion increased from 11 % to

17%, a proportionately slight decline in the texture score was observed and a significantly

(P<0.05) lower score in T4 group was recorded when compared with the control group. The drop

in texture score could be attributed to the very soft and loose consistency and less chewy texture

of the products resulting from the higher moisture content, thereby, affecting its palatability. The

same trend was observed on 5th, 10th and 15th day between T1 and T4 under both aerobic and

57

vacuum packaging system. On the other hand, T2 was found with scores that were non

significant when compared to control group up to 15th day, both under aerobic and vacuum

packaged. The group T3 scored significantly lower scores on 10th day (vacuum) and 15th day

(aerobic and vacuum) as compared to that of control group.

The effect on texture of control group at all point of storage period was notably non-

significant. In contrary, all the blood incorporated groups (aerobic packaged) were recorded with

a significantly lower scores by the end of 15th day of storage, which, however, was not observed

in vacuum packaged groups. In addition, it was noted that the product stored under vacuum

condition exhibited a better texture score as compared to the products preserved under aerobic

condition.

Table 6.02.05: Overall acceptability scores (Mean±SE) of chicken nuggets at different blood

levels under aerobic or vacuum packaged condition (4±1°C)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic B7.00b±0.00 B6.93b±0.07 C6.50ab±0.19 B6.14a±0.26

5

Aerobic AB6.79b±0.15 B6.64ab±0.14 BC6.29ab±0.26 B6.00a±0.11

Vacuum B7.00b±0.00 B6.71ab±0.29 BC6.43ab±0.17 B6.07a±0.23

10 Aerobic AB6.50b±0.29 AB6.21ab±0.15 ABC5.93ab±0.25 AB5.50a±0.19

Vacuum AB6.71b±0.15 AB6.36ab±0.18 ABC6.21ab±0.18 AB5.79a±0.31

15

Aerobic A6.14b±0.18 A5.79ab±0.21 A5.50ab±0.31 A5.14a±0.24

Vacuum AB6.43b±0.13 A6.00ab±0.11 AB5.71ab±0.21 AB5.50a±0.19

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

The panelists conferred the highest overall acceptability score of 7.00±0.00 to the control

group on 0 day (Table 6.02.05). The T4 group with the maximum level of blood (17%) was

recorded with significantly (P<0.05) lower score of 6.14±0.26 as compared with the control

group. However, it is noteworthy that the overall acceptability was scored at 6.14±0.26 under 7-

point Hedonic scale, which is still a good score of acceptance. The same trend followed between

58

T4 and T1 groups, both aerobic and vacuum packaged, up to 15 day of storage. On the other

hand, no significant difference was observed between T1, T2 and T3 groups, aerobic and

vacuum alike, during the storage period.

The overall acceptability score of the nuggets at all levels of blood incorporation and

packaging systems declined gradually with time but was markedly significant (P<0.05) by the

15th day in all the aerobic packaged groups. On comparison between the two packaging methods,

the vacuum system exhibited a slightly better overall acceptance than its aerobic packaged

counterpart all through the storage period, revealing its ability to retain sensory qualities for a

longer duration.

From the overall acceptance score of chicken nuggets, the 11% blood group has been

graded under “Very good to Excellent” according to the 7-point Hedonic scale among the blood

incorporated groups suggesting the acceptability of its sensory quality. This reflects the

possibility of using whole blood up to 11% for nuggets without serious effect on sensory

qualities and stored up to 10 days at 4±1°C, preferably under vacuum packaging method, though

aerobic packaging also still holds good for consumption up to 10 days.

Storage stability studies:

a) Room temperature:

It was observed that the chicken meatballs and nuggets packed under aerobic and vacuum

atmosphere and stored at room temperature (35-38°C) exhibited signs of spoilage after 24 hours

of its preparation. The presence of slime and off odour was evident. Further studies were not

conducted as the meat products were spoiled which is a distinct indication of room temperature

being unfavourable for storage of meat products and unsafe for consumption.

b) Refrigerated temperature:

Under refrigerated temperature (4±1°C), physico-chemical studies viz., pH, TBA, Tyrosine,

Water holding capacity and microbial studies viz., Total plate count, yeast & mould count,

presence of salmonella, if any, were carried out for both the meat products packaged under the

aerobic or vacuum nodule to monitor the storage stability. The results and observations are

presented below.

59

pH:

The pH is an important physico-chemical property of meat which has an imperative

influence on other quality traits viz., emulsifying capacity, emulsion stability, cooking loss,

flavour, juiciness, texture and drip loss. Water holding capacity and emulsifying ability of meat

are both affected when the meat or its product undergo changes in pH. The pH, being an

important measure to estimate relative acidity or alkalinity of meat and meat products, is

indicative of the potential storage life of the meat/meat products. The mean pH values of

aerobically and vacuum packaged chicken meatballs and chicken nuggets stored at refrigerated

temperature are shown in Table 7.01 and 7.02, respectively.

Table 7.01: pH values of chicken meatballs at different blood levels under aerobic or

vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic A6.22a±0.002 A6.24b±0.002 A6.26c±0.002 A6.27c±0.003

5

Aerobic A6.23a±0.002 A6.25b±0.002 A6.27c±0.002 B6.29d±0.002

Vacuum A6.22a±0.002 A6.25b±0.002 A6.26b±0.002 AB6.28c±0.002

10

Aerobic B6.27a±0.004 B6.29b±0.01 B6.30b±0.004 C6.32c±0.01

Vacuum B6.26a±0.002 B6.28b±0.002 B6.30c±0.01 C6.31c±0.01

15

Aerobic D6.36a±0.01 C6.37a±0.002 D6.37a±0.002 E6.39b±0.002

Vacuum C6.29a±0.002 B6.29a±0.004 C6.34b±0.002 D6.35b±0.004

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

It was observed that the pH values of freshly prepared chicken meatballs significantly

(P<0.05) increased with the increase in the level of blood incorporation (Table 7.01.01). On 0

day, the pH value of the control group was recorded at 6.22±0.002 while the highest pH value

was recorded in the T4 group (6.27±0.003) where the maximum blood inclusion was made

60

(10%). The pH of all the blood incorporated groups were significantly higher (P<0.05) than the

control group at all point of storage, except on 15th day. The increase in pH values in the blood

incorporated groups are mainly due to the addition of blood which has higher pH (7.6). In the

control group the pH was influenced by the inherent pH of meat which ranges from 5.5 to 6.0.

Higher pH in meat products in comparison to fresh meat is attributed to higher degree of

oxidation and loss of free acidic groups of meat protein upon cooking (Lawrie, 1998; Lingaiah

and Reddy, 2001)

During the storage period at 4±1°C, the pH values gradually increased from 0 day

onwards. A significant increase was recorded on 5th day for aerobic packaged T4 group while no

significant increase were observed in others groups under aerobic and vacuum condition. After

the 5th day of storage, significantly (P<0.05) higher pH were observed in all the groups. Till 10th

day of storage, no significance difference was recorded between the two packaged systems, in

each group. This may be due to the oxidation of fatty acids during storage and a concomitant

increase of bacteria which release metabolites during their metabolism (Jay, 1996). The results

are in accordance with several authors, who also reported increase in pH values of meat products

stored under refrigeration (Reddy, 2008; Kim et. al., 2014; Marcinkowska-Lesiak et. al., 2016).

In addition, it was interesting to note that the change of pH in the vacuum packed meatballs

was lower than that of aerobic packed environment. This is in agreement with Sahoo and

Anjaneyulu (1997) who reported a significantly (P<0.05) higher pH values of vacuum packaged

buffalo meat nuggets than control aerobic packaged. It was evident that storage time and

packaging method exerted considerable effects on pH of the chicken meatballs.

Table 7.02: pH values of chicken nuggets at different blood levels under aerobic or vacuum

packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic A6.36a±0.01 A6.45b±0.01 A6.47bc±0.004 A6.50c±0.02

5

Aerobic B6.40a±0.01 A6.46b±0.01 AB6.48b±0.003 AB6.53c±0.002

Vacuum A6.38a±0.002 A6.45b±0.004 AB6.48bc±0.002 A6.50c±0.002

61

10

Aerobic B6.41a±0.01 B6.51b±0.004 BC6.51b±0.02 B6.54b±0.004

Vacuum B6.40a±0.01 A6.46b±0.004 AB6.49b±0.01 AB6.53c±0.002

15

Aerobic B6.43a±0.002 B6.51b±0.01 C6.53bc±0.002 B6.55c±0.01

Vacuum B6.41a±0.01 A6.47b±0.004 BC6.51c±0.01 B6.54c±0.002

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

The pH value in freshly prepared chicken nugget was 6.36±0.01 in the control group. The

value increased significantly (P<0.05) in all the treated groups, i.e., T2, T3 and T4 (Table 7.02).

The same trend was seen throughout the storage period. The higher pH clearly shows the

influence of the added blood in the nuggets. By and large, the nature of change in pH values

under different treatment groups and the entire storage period was similar to that of meatballs.

With storage, the pH values gradually increased and the significant (P<0.05) change was

observed on 10th and 15th day in both the packaged groups. The vacuum packaged nuggets

showed numerically lower pH scores than its aerobic counterparts in all the groups at all point of

storage.

Among the blood treated groups, T2 group was recorded with the lowest pH value in both

packaging modules which is a desirable trait for longer storage stability. Nonetheless, storage up

to 10th day is preferred in the treated groups considering the high pH nature of the blood.

Thiobarbituric acid (TBA) value

The storage stability of any lipid containing food product is estimated by the extent of oxidative

rancidity developed in it. Rancidity of fat in stored foods is defined as the development of stale

off flavour as a result of oxidation of unsaturated fatty acids. Thiobarbituric acid value measures

the carbonyl residues in the form of malondialdehyde (MDA) formation which is a breakdown

product of peroxidised polyunsaturated fatty acid resulting from lipid peroxidation in the

product. The rate of this change depends on initial bacterial load, physical and biochemical

change, availability of oxygen, temperature of storage and muscle composition.

62

The mean TBA values of chicken meatballs as influenced by incorporation of 5%, 7.5%

and 10% blood, storage period and packaging methods are given in Table 8.01. On 0 day, a

significant (P<0.05) rise in the TBA values were observed with the increasing blood levels in the

meatballs. The TBA value for T1 group was recorded at 0.26±0.01 mg MDA/kg which was

significantly (P<0.05) lower than the T2, T3 and T4 groups. This trend was followed at all point

of storage period regardless of aerobic or vacuum packaged system. This demonstrated a higher

lipid oxidation in the blood incorporated groups when compared to the control group which may

be due to more iron content released from heme pigment on heating and iron is one of the major

catalysts for lipid oxidation (Love and Persson 1974; Igene et.al., 1979).

Table 8.01: TBA values (mg malondialdehyde/kg) of chicken meatballs at different blood

levels under aerobic or vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic A0.26a±0.01 A0.32b±0.01 A0.35c±0.004 A0.38d±0.01

5

Aerobic C0.49a±0.01 C0.59b±0.004 C0.64c±0.01 C0.69d±0.01

Vacuum B0.47a±0.001 B0.55b±0.002 B0.60c±0.001 B0.64d±0.002

10

Aerobic E0.72a±0.002 E0.86b±0.002 E0.93c±0.002 E1.02d±0.01

Vacuum D0.68a±0.01 D0.81b±0.01 D0.89c±0.004 D0.95d±0.003

15

Aerobic G1.03a±0.001 G1.17b±0.002 G1.23c±0.002 G1.33d±0.002

Vacuum F0.92a±0.002 F1.11b±0.003 F1.17 c ±0.002 F1.26d±0.002

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

A progressive and significant (P<0.05) increase in the TBA values was noticed during the

storage period irrespective of the blood levels or packaging methods. Chowdhury et. al. (2015),

Das et al. (2013), Kumar and Tanvar (2011) and Chidanandaiah et al. (2009) also reported a

similar increase in TBARS values upon storage of different meat products.

63

It was also noted that the meatballs packaged under vacuum condition showed

numerically lower TBA value as compared to those under aerobic packaging. This was

consistent in all the treated groups. In agreement to our findings, Bhoyar et al., (1997) reported

that vacuum packaged chicken steaks exhibited lower TBA values than their air packaged

counterparts throughout the storage period and also recorded a significant increase in TBA

values during storage in both vacuum and aerobically heat sealed packaging systems. It may be

attributed to aerobic packaging of product and oxygen permeability of packaging material that

led to faster lipid oxidation (Brewer et al., 1992; Nag et al., 1998). However, in the present

study, the TBA values were lower than threshold value of 2.0 mg MDA/kg (Greene and Cumuze,

1982) even by day 15, which otherwise would produce detectable off odours or flavor. Among

the blood incorporated groups, T2 group recorded with lowest TBA values at all point when

compared to T3 and T4. However, if a higher level of blood inclusion was to be opted, T3 group

with 7.5% blood also exhibits values at a safe range up to 15 day of storage which makes it

acceptable level. Nevertheless, storage up to 10 days may be preferred over 15 days due to the

higher pH, protein and moisture content of the meat product under consideration.

Table 8.02: TBA values (mg malondialdehyde/kg) of chicken nuggets at different blood

levels under aerobic or vacuum packaged condition (Mean±SE)

Storage

period (Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic A0.29a±0.002 A0.41b±0.001 A0.45c±0.001 A0.52d±0.001

5

Aerobic C0.54a±0.001 C0.72b±0.001 C0.78c±0.01 C0.88d±0.003

Vacuum B0.50a±0.002 B0.68b±0.01 B0.74c±0.002 B0.83d±0.001

10

Aerobic E0.80a±0.002 E1.03b±0.002 E1.11c±0.001 E1.24d±0.002

Vacuum D0.71a±0.01 D0.94b±0.002 D1.02c±0.003 D1.15d±0.002

15

Aerobic G1.07a±0.002 G1.34b±0.002 G1.41c±0.002 G1.50d±0.003

Vacuum F0.98a±0.002 F1.25b±0.001 F1.32c±0.002 F1.41d±0.004

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

64

The results of mean values of TBA of chicken nuggets as influenced by incorporation of

11%, 14% and 17% blood, storage period and packaging methods are given in the Table 8.02.

The TBA value of freshly prepared chicken nuggets for the control group was recorded

the lowest, i.e., 0.29±0.02 mg MDA/kg, while a significantly (P<0.05) higher TBA values were

recorded in all the blood incorporated groups, with T4 group exhibiting the highest value

(0.52±0.01). The nature of response of TBA with the increasing blood levels in the meat

emulsion indicated the increase in lipid oxidation due to the presence of more heme iron, a major

catalyst of lipid oxidation. The findings are similar to chicken meatballs.

The TBA values of chicken nuggets also followed a significantly (P<0.05) increasing

trend from day 0 to 15 in case of all the groups and packaging systems (aerobic and vacuum).

Also, the vacuum packaged nuggets exhibited significantly (P<0.05) lower TBA values as

compared to those under aerobic packaging. This might be attributed to aerobic packaging of

product and oxygen permeability of packaging material (Brewer et al. 1992) that led to lipid

oxidation as discussed above. Despite the increase in the TBA values with storage time, the

values did not exceed the threshold value of 2.0 mg MDA/kg (Greene and Cumuze, 1982) even

at day 15, showing no indication of oxidative rancidity. On comparison among the blood

incorporated groups, T2 recorded with the lowest TBA values at all point of storage when

compared to T3 and T4. This makes T2 group, with 11% blood is preferable among the treated

groups.

Tyrosine value

Tyrosine value is used as method of detecting microbial spoilage in muscle foods (Jay,

1987). It measures tyrosine and tryptophan present in the protein extract of meat which is taken

as an indicator of proteolysis (Strange et al., 1977). Mean tyrosine values obtained at different

storage intervals in refrigeration (mg/100g) for aerobically and vacuum packaged chicken

meatballs and chicken nuggets incorporated with different levels of blood are presented in the

Table 9.01.01 and 9.02.01, respectively.

65

Table 9.01: Tyrosine values (mg/100g) of chicken meatballs at different blood levels under

aerobic or vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic A26.48a±0.02 A27.69b±0.1 A30.78c±0.05 A32.46d±0.05

5

Aerobic A26.66a±0.03 A27.99b±0.06 A31.01c±0.05 A32.73d±0.03

Vacuum A26.61a±0.02 A27.85b±0.13 A30.96c±0.12 A32.67d±0.05

10

Aerobic D30.77a±0.17 B32.39b±0.05 B35.52c±0.03 B37.35d±0.12

Vacuum A26.74a±0.07 A28.00b±0.04 A31.15c±0.11 A32.88d±0.02

15

Aerobic C39.06a±0.04 C40.77b±0.10 C43.98c±0.39 C46.08d±0.19

Vacuum B31.00a±0.17 B32.32b±0.05 B35.61c±0.11 B37.40d±0.11

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

On 0 day, the tyrosine value of the chicken meatballs exhibited an increasing trend from

26.48±0.02 (control) to 32.46±0.05 (10% blood). The change observed was a significant

(P<0.05) increase in the tyrosine value with the increase in blood inclusion level in the meat

emulsion (Table 9.01). Similar development was evident at all point of storage period regardless

of aerobic or vacuum packaged system. This may be due to the higher pH and protein content in

the blood incorporated groups leading to proteolysis and more protein degradation influencing

the tyrosine value. During storage, the tyrosine value significantly (P<0.05) increased on 10 day

in all the aerobic packaged groups; however, considerable change was not observed in vacuum

packaged groups.

On the 15th day, the increase (P<0.05) in tyrosine values was noteworthy in all the groups

packaged under both aerobic and vacuum. In concordance to our findings, Biswas et al. (2011)

also reported a highly significant increase in tyrosine value with increase in storage period at

refrigeration temperature. Pearson (1968) stated that tyrosine value increased with the storage

until deamination of amino acids limited the formation of free amino acids.

66

Table 9.02: Tyrosine values (mg/100g) of chicken nuggets at different blood levels under

aerobic or vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic A29.38a±0.03 A33.35d ±0.03 A32.58c±0.18 A30.80b±0.23

5

Aerobic A29.56a±0.07 A33.68d±0.08 A32.82c±0.18 A31.08b±0.24

Vacuum A29.52a±0.08 A33.50d±0.04 A32.77c±0.20 A31.01b±0.21

10 Aerobic C33.67a±0.05 C39.08d±0.10 C38.36c±0.03 B35.67b±0.04

Vacuum B30.01a±0.04 B34.02d±0.08 B33.24c±0.07 A31.22b±0.04

15

Aerobic E41.25a±0.08 D49.69d±0.06 E46.56c±0.07 C42.47b±0.04

Vacuum D35.52a±0.03 C39.40d±0.09 D38.89c±0.09 B36.06 b±0.10

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

In chicken nuggets, the tyrosine value on 0 day was recorded at 29.38±0.03, 33.35±0.03,

32.58±0.18 and 30.80±0.23 mg/100mg in T1, T2 T3 and T4 groups, respectively (Table 9.02).

All the blood incorporated groups showed significantly (P<0.05) higher tyrosine value than their

control counterpart. The increase in the values is attributable to the added blood which has a high

pH (7.6) influencing on the tyrosine value. Also, all the groups exhibited significantly (P<0.05)

different tyrosine values on comparison among the groups and this trend was observed up to 15

day regardless of the packaging method. It was also noteworthy that with the increment in

addition of blood, a fall in tyrosine value was recorded in groups, T3 and T4 when compared to

T2. This may be in correlation with the protein content where T2 exhibits the highest level in

comparison with its treated counterparts.

The tyrosine value behaved similarly to that of chicken meatballs in respect to different storage

period under both the packaging conditions.

Tyrosine value is an indicator of proteolysis and protein degradation which have some

degree of correlation with pH and standard plate count of the product (Eyas ahmed, 2007).

Therefore, the longer exposure time might have allowed the meat product to undergo autolysis or

67

bacterial proteolysis. However, in all cases it was observed that vacuum atmosphere had

considerable effect on lowering the rate of proteolysis which was reflected in lower tyrosine

value in comparison to its aerobic counterpart.

Water holding capacity (WHC)

The water holding capacity(WHC) of meat products is a very important quality attribute

which has an influence on product yield and is also important in terms of eating quality (Cheng

and Sun, 2008). It essentially affects the appearance before cooking, behavior during cooking

and juiciness on mastication (Lawrie, 2006). Water holding capacity is a desirable character in

relation to its palatability and functional property (Reddy, 2008). The water holding capacity of

chicken meatballs and chicken nuggets packaged under aerobic and vacuum condition are

displayed in Table 10.01and Table 10.02, respectively.

Table 10.01: WHC (ml/100g) of chicken meatballs at different blood levels under aerobic or

vacuum packaged condition (Mean±SE)

Storage

period (Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic G41.44a±0.14 F48.18b±0.11 F50.22c±0.14 E51.56d±0.14

5

Aerobic E40.09a±0.17 E47.02b±0.09 E48.18c±0.11 C49.51d±0.11

Vacuum F40.80a±0.22 F47.82b±0.11 D49.42c±0.17 D50.40d±0.11

10

Aerobic C38.40a±0.11 C43.56b±0.14 B45.42c±0.17 B47.20d±0.17

Vacuum D39.11a±0.14 D46.22b±0.14 C47.47c±0.17 C49.33d±0.14

15

Aerobic A36.09a±0.09 A41.24b±0.17 A43.20c±0.09 A45.51d±0.11

Vacuum B37.60a±0.11 B44.18b±0.11 B45.33c±0.14 D47.29d±0.11

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

68

The freshly prepared chicken meatballs recorded WHC values of 41.44±0.14,

48.18±0.11, 50.22±0.14 and 51.56±0.14 in T1, T2, T3 and T4 groups, respectively (Table

10.01). The WHC of the chicken meatballs were found to be significantly (P<0.05) higher in all

the blood treated groups when compared to control group. The replacement of lean meat with

whole blood at increasing levels resulted in an increase in moisture level which resulted in higher

WHC as the behavior of water itself is an additive and the ratio of water to meat affects the

overall WHC of the meat emulsion (Warner, 2017). Additionally, higher pH and strong emulsion

formation might also have contributed towards higher water holding capacities in the meat

product (Chowdhury et.al, 2015). Throughout the storage period, the WHC was observed to be

significantly higher in the blood incorporated groups than the control. The value differed

significantly (P<0.05) among the blood treated groups.

The mean WHC values decreased significantly (P<0.05) with increase in storage period

irrespective of the groups or packaging methods. This might be due to the denaturation of

myofibrillar proteins and loosening up of the microstructure of muscles allowing more water to

be entrained (Nagamallika et al., 2006). The nature of response of WHC on packaging method

throughout the storage period revealed that the vacuum packaged atmosphere helped the

meatballs in retaining water better in comparison to aerobic packaged group. Higher the WHC,

better is the quality of the meat product.

Table 10.02: WHC (ml/100g) of chicken nuggets at different blood levels under aerobic or

vacuum packaged condition (Mean±SE)

Storage

period (Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic E42.22a±0.14 E51.11b±0.14 F54.49c±0.11 F59.73d±0.11

5

Aerobic D41.07a±0.11 D49.60b±0.11 D52.18c±0.23 D57.33d±0.14

Vacuum E41.78a±0.14 E50.49b±0.11 E53.42c±0.17 E58.49d±0.23

10

Aerobic C40.00a±0.24 B46.40b±0.11 B49.24c±0.17 B53.69d±0.22

Vacuum C40.36a±0.17 C48.44b±0.20 C51.38c±0.11 C56.00d±0.14

69

15

Aerobic A36.36a±0.17 A43.47b±0.17 A46.31c±0.17 A51.38d±0.11

Vacuum B38.40a±0.27 B46.40b±0.11 B49.16c±0.11 B54.22d±0.14

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

On 0 day, the freshly prepared chicken nuggets recorded a significantly (P<0.05) higher

mean WHC values for T2 (51.11±0.14), T3 (54.49±0.11) and T4 (59.73±0.11) groups than the

control group T1 (42.22±0.14). Significant difference in the WHC values were also observed

among the blood treated groups. This trend was exhibited throughout the storage period. From

the above results, it is noteworthy that WHC was affected by the level of blood incorporated in

the chicken meat nugget formulations. The nature of response of WHC in respect to blood levels

showed (Table 10.02) an increased WHC value with higher blood level which may be mainly

due to higher moisture level, higher alkalinity and strong emulsion formation. The WHC

behavior of chicken nuggets with respect to storage period and packaging method were found to

be similar to chicken meatballs.

Microbial studies

The behavior of microorganisms in foods is governed by the constraints through a variety

of environmental and ecological factors. These include water activity, pH, chemical composition,

presence of natural or added antimicrobial agents and storage temperature as well as the

processing factors such as heat treatment and physical manipulation (Hobbs, 1986).

Total Plate Count (TPC)

The chicken meatballs and chicken nuggets with different blood levels were subjected for

Total Plate Count (TPC log10 cfu/g) at day zero and subsequently at 5 th, 10th and 15th day of

storage at 4±1°C. The mean values of Total Plate Count (TPC) of aerobically and vacuum

packaged chicken meatballs and chicken nuggets containing different level of blood are

presented in Table 11.01 and Table 11.02, respectively.

70

Table 11.01: TPC (cfu/g) of chicken meatballs at different blood levels under aerobic or

vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (5% blood)

TREATMENT

T3 (7.5% blood)

TREATMENT

T4 (10% blood)

0 Aerobic ND ND ND ND

5

Aerobic ND ND ND ND

Vacuum ND ND ND ND

10

Aerobic B3.07a±0.04 B3.20ab±0.04 B3.25b±0.03 A3.32b±0.01

Vacuum ND ND ND ND

15

Aerobic C3.45a±0.01 C3.51ab±0.02 C3.57ab±0.01 B3.65b±0.02

Vacuum A2.74a±0.09 A2.92b±0.05 A3.06bc±0.07 A3.18c±0.03

(ND= Not detected);

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ significantly (P<0.05)

There was no evidence of viable microbial growth in the chicken meatballs up to 5th day

of storage in all the treated groups as well as two packaging system. This shows the sanitary and

hygiene level maintained during processing and insignificant post processing contamination. The

analysis on the data showed that there was an impact on the TPC count due to addition of blood,

period of storage and packaging atmosphere which was observed from the 10th day of storage.

There was an apparent growth of microorganisms by day 10 in the aerobically packaged

meatballs in all groups and the count was significantly (P<0.05) higher in T3 and T4 when

compared to control but no significance difference was observed among the blood treated

groups. A significantly higher TPC count in all blood treated groups (aerobic and vacuum) was

observed when compared to its control counterpart. Higher TPC count may be attributed to the

effect of blood addition which contributed higher level of moisture, protein, amino acids and pH

and the availability of oxygen which created a more conducive medium for growth of

microorganisms than the control group.

71

On the 15th day of storage, a significant (P<0.05) increase in the TPC count was observed

in all the groups under aerobic as well vacuum packaged system. A similar result was also

reported by Kumar et. al. (2015) where the microbial growth increased with the time of storage.

It is noteworthy that the development of mesophiles occurred more quickly in the product stored

using aerobic packaging than in the vacuum-packed product. The higher oxygen permeability of

the first packaging module may have been the main reason for the rapid microbial growth. This

also shows that vacuum packaging had a better shelf–life when compared to aerobic packaging

under refrigerated conditions.

In the present study, TPC count of control and treatment groups of chicken meatballs

were within the permissible level (log 103 cfu/g of sample) of Food Safety and Standards (Food

products standards and additives) Regulations, 2011, India in cooked meat products, even on 15th

day of storage. There was no evidence of spoilage of meat products and remained safe for

consumption. Therefore, using of blood at 7.5% level in chicken nuggets preparation is safe up to

15 days of refrigerated storage, provided strict sanitary measures is practiced to avoid pre- and

post-processing contaminations. Blood being a good source of nutrients provides a good medium

for microbial growth, 10 day storage period may be considered under vacuum packaging to

extend the shelf life of the product.

Table 11.02: TPC (cfu/g) of chicken nuggets at different blood levels under aerobic or

vacuum packaged condition (Mean±SE)

Storage

period

(Days)

Packaging

condition

CONTROL

T1 (0% blood)

TREATMENT

T2 (11% blood)

TREATMENT

T3 (14% blood)

TREATMENT

T4 (17% blood)

0 Aerobic ND ND ND ND

5

Aerobic ND ND ND ND

Vacuum ND ND ND ND

10

Aerobic B3.02a±0.04 A3.33c±0.02 A3.25bc±0.02 A3.23b±0.03

Vacuum ND ND ND ND

72

15

Aerobic C3.48a±0.02 B3.71c±0.02 B3.64bc±0.01 B3.61b±0.01

Vacuum A2.92a ±0.05 A3.33c±0.02 A3.27bc±0.03 A3.21b±0.05

(ND= Not detected);

Means bearing same superscripts column-wise (Lower case) or row-wise (Upper case) do not differ

significantly (P<0.05)

The chicken nuggets with different blood levels packed under aerobic and vacuum

method showed similar behavior to that of chicken meatballs in respect to packaging conditions

and storage time period for all four treatment groups. The TPC count was significantly (P<0.05)

higher in T2 and T3 groups than the control and T4 groups. The TPC value between T2 and T3

were comparable.

The growth rate of microorganism was also found faster in blood incorporated groups

and those packaged under aerobic with storage time. The increasing trend of microbiological

count in present study was similar to the findings were reported by Kumar et al. (2007) in

chicken meat patties. Among the blood treated groups, the group with 11% blood was recorded

with the highest TPC count; however, it is far below the permissible limit (log 103 cfu/g of

sample) in cooked meat products, even on 15th day and the level may be considered for inclusion

in meat emulsion for nuggets preparation.

Yeast and Mould count

A food safety concern with regard to surface growth of moulds on any meat or meat

products is the mycotoxin production. Many studies have reported the development of these

microorganisms in meat products stored under refrigeration, especially under aerobic conditions

(Parra et al., 2010, Samelis & Georgiadou, 2000; Santos et al., 2005). In the present study,

analysis of yeast and mould for chicken meatballs and nuggets were performed to check for

potential growth of these microorganisms during the storage period under refrigerated

temperature. The Yeast and Mould count observed for chicken meatballs and chicken nuggets

are displayed in the Table 12.01 and Table 12.02, respectively.

73

Table 12.01: Yeast & Mould Count (cfu/g) (Mean±SE) of chicken meatballs at different

blood levels under aerobic or vacuum packaged condition (4±1°C)

Day

CONTROL T1 (0% blood )

TREATMENT T2 (5% blood )

TREATMENT T3 (7.5% blood)

TREATMENT T4 (10.% blood)

Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum

0 ND ND ND ND ND ND ND ND

5 ND ND ND ND ND ND ND ND

10 ND ND ND ND ND ND ND ND

15 ND ND ND ND ND ND ND ND

(ND= Not detected)

The yeast and mould counts were not detected for the chicken meatballs groups both

under aerobic or vacuum packaging module up to 15th day of storage at refrigerated temperature

(4±1°C). This revealed that there was no post processing contamination and no potential growth

up to 15 days of refrigerated storage in both aerobic and vacuum packaging module.

Table 12.02: Yeast & Mould Count (cfu/g) (Mean±SE) of chicken nuggets at different blood

levels under aerobic or vacuum packaged condition (4±1°C)

Day

CONTROL T1(0% blood)

TREATMENT T2 (11% blood)

TREATMENT T3 (14% blood)

TREATMENT T4 (17%blood)

Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum

0 ND ND ND ND ND ND ND ND

5 ND ND ND ND ND ND ND ND

10 ND ND ND ND ND ND ND ND

15 ND ND ND ND ND ND ND ND

(ND= Not detected)

Similar to chicken meatballs, the yeast and mould counts were also not detected in the

chicken nuggets irrespective of blood levels or packaging method up to 15 day of refrigerated

storage which indicated the high sanitary standard maintained during the processing and also no

evidence of post processing contamination. The shelf life of chicken nuggets was found to be

intact up to 15 days of refrigerated storage in both aerobic and vacuum packaging modules.

74

Presence or absence of Salmonella

Poultry meat is one of the most consumed and expanding globally traded meat products

(Antunes et. al., 2016). Salmonella infections are of major public health concern worldwide as

this pathogen causes food-borne disease in humans (Chen et. al., 2010). The presence of

Salmonella in poultry meat and poultry meat products occurs primarily due to cross-

contamination and undercooking (Luber, 2009). The incidence of Salmonella in meat or meat

products shows the need for close supervision of processing and sanitation practices (Weissman

and Carpenter, 1969). The detection of this pathogen in poultry meat and its products, both at the

production level and before consumption, can play a significant role in the prevention of food-

borne salmonellosis (Temelli et. al., 2012). In the present study, test was conducted for the

presence of salmonella, if any, in the chicken meatballs and chicken nuggets and the results are

shown in Table 13.01 and 13.02, respectively.

Table 13.01: Incidence of Salmonella in chicken meatballs at different blood levels under

aerobic or vacuum packaged condition (4±1°C)

Day

CONTROL T1 (0% blood)

TREATMENT T2 (5% blood )

TREATMENT T3 (7.5% blood)

TREATMENT T4 (10% blood)

Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum

0 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

5 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

10 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

15 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

(+ = presence of Salmonella in one sample; - = absence in one sample; number of replicates=5)

Table 13.02: Incidence of Salmonella in chicken nuggets at different blood levels under

aerobic or vacuum packaged condition (4±1°C)

Day

CONTROL T1 (0% blood)

TREATMENT T2 (11% blood)

TREATMENT T3 (14% blood)

TREATMENT T4 (17%blood)

Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum Aerobic Vacuum

0 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

5 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

10 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

15 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

(+ = presence of Salmonella in one sample; - = absence in one sample; number of replicates=5)

75

The incidence of Salmonella was not detected in chicken meatballs and chicken nuggets

incorporated with different levels of blood during the entire refrigerated storage (4±1°C) period

of 15 days under both aerobic and vacuum packaging conditions for all treatment groups (Table

13.01 and 13.02). This indicated that proper sanitation practices were followed during processing

and the products were safe for consumption.

The enrichment of spent hen meat products with iron as envisioned through the

utilization of slaughter by-products like blood is found promising from the findings of the

present study. The inclusion of whole blood for preparation of meat balls was found to be most

suitable at 7.5% which showed 99.61% high iron and 11.94% more protein contents and 11.67%

lower fat without compromising in the sensory qualities and a good response of acceptance by

the taste panelists. Likewise, inclusion of 11% blood was acceptable in regards to sensory

qualities of nuggets which imparted 184.49% more iron, 7.07% higher protein, correspondingly

19.75% lower dietary fat in the product.

76

CHAPTER V

SUMMARY AND CONCLUSION

The present study was aimed to utilize spent hen for production of chicken meatballs and chicken

nuggets enriching it with iron through the use of chicken blood. Initially, the feasibility of

incorporating the appropriate level of blood for production of meatballs and nuggets were

explored through several pilot studies. Whole blood was used to ensure its uniform mixing in the

meat emulsion. The standardized basic recipe for the control, respective to the chicken nuggets

and chicken meatballs were prepared. For the treated chicken nuggets, lean meat was replaced

with whole blood at 11%, 14% or 17%. On the other hand, for chicken meatballs, lean meat was

replaced with whole blood at 5%, 7.5% or 10%. The chicken meatballs and chicken nuggets

were subjected to several compositional and organoleptic studies. The blood incorporated meat

products showed a significant (P<0.05) increase in the iron content to the extent of 154.56% in

meatballs and 264.72% in nugget at highest level on comparison to their control counterparts.

Simultaneously, the protein content in both the products were significantly (P<0.05) increased.

On the other hand, both fat and cholesterol contents in meatballs and nuggets were significantly

(P<0.05) reduced. On sensory evaluation of the chicken meatballs and chicken nuggets, it was

observed that an addition of whole blood up to 7.5% to chicken meatballs and 11% to chicken

nuggets did not affect their overall acceptability wherein the panelist chose ‘Excellent to Very

good’ score. This revealed the acceptability and possibility of using blood at the said levels

without any detrimental effect on sensory qualities. The meatballs and nuggets retain their

sensory qualities up to 15 days of storage at 4±1°C. The various storage stability studies of the

meat products were undertaken for refrigerated temperature (4±1°C) at an interval of 5 days up

to 15 days. The packaging module was limited to only aerobic and vacuum since the modified

atmospheric packaging equipment became non-functional. The proposed studies under room

temperature were discontinued because of evident spoilage after 24 hours post-preparation of the

meat products. It is an indication of room temperature being unfavourable for storage of meat

products and unsafe for consumption.

The pH values of freshly prepared chicken meatballs and chicken nuggets increased

proportionately with the increase in the level of blood incorporation which might be due to the

influence of higher blood pH (7.6). During storage period at 4±1°C the pH value gradually

77

increased with the time, starting from the 0 day to 15th day, irrespective of the blood

levelincorporated or the packaging methods. The thiobarbituric acid (TBA) values of chicken

meatballs and chicken nuggets were found to rise significantly (P<0.05) with the increase in

blood levels demonstrating a higher lipid oxidation than their control counterparts, which may be

due to the higher catalytic impact of iron on lipid oxidation. The TBA values increased

progressively with storage time. All the blood incorporated groups of chicken meatballs and

nuggets showed significantly (P<0.05) higher tyrosine values which might be due to the higher

pH level and richer protein content in the blood. The tyrosine value increased proportionately

with the storage time in all the groups under both packaging system owing to the intrinsic

changes. The water holding capacity (WHC) of the chicken meatballs and chicken nuggets were

found to be significantly (P<0.05) higher in all the blood treated groups. The replacement of lean

meat with whole blood at increasing levels, higher pH and strong emulsion resulted in elevated

moisture level and subsequently higher WHC. However, with increase in storage period, decrease

in WHC was observed resulting from denaturation of myofibrillar proteins and loosening up of

muscle-microstructures allowing more water to be entrained.

No microbial growth was observed in any of the samples till 5th day but was subsequently

noted with the extension of storage period. However, the total plate count (TPC) of all the groups

were within the permissible level (log 103 cfu/g of sample), both under aerobic and vacuum

packaging. Also, neither yeast and mould nor incidence of Salmonella were detected in both the

meat products (aerobic and vacuum packaged) throughout the storage period. It was interesting

to note that during the entire study period the vacuum packaging gave better results than its

aerobic counterpart, in terms of sensory qualities as well as physico-chemical and microbial

status.

From the findings of the study, it was concluded that chicken meatballs with 7.5% and

chicken nuggets with 11% blood incorporation could increase the iron content to the extent of

99.62 and 184.49 %, respectively with remarkable protein elevation. Most importantly, these two

levels were highly acceptable in regard to sensory qualities.

The major issue observed in the study was shelf life of the meat products under refrigeration

temperature (4±1°C). The meat products remained safe for consumption without any affect in the quality

up to 10 days. It is recommended that the meat products may be stored under freezing temperature (18-

21°C) for better shelf life and longer storage duration (3 months). Also, vacuum packaging is

78

recommended over aerobic packaging as discussed above.

In an effort to transfer of the developed technology to end users, leaflets in English and

Assamese were prepared as hands on the technology to the end users. Correspondingly, a Small-

Scale Poultry Product Preparation Model Unit was developed for the unemployed youths and

two Entrepreneurial training programmes were organized involving youths from different districts of

Assam.

79

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89

ANNEXURE- I

SCORE CARD FOR TASTE PANEL EVALUTION

Product: Date:

Name:

Introduction :

The product should be judged according to the following criteria. Any additional

comments would be highly appreciated and may be written in the space provided for the

purpose.

Scores under Hedonic scale Excellent 7

Very good 6

Good 5

Fair 4

Poor 3

Very poor 2 Extremely poor 1

Sample Colour Flavour Juiciness Texture Overall

Acceptance

A

B

C

D

Remarks: Signature

90

ANNEXURE –II

SMALL - SCALE PREPARATION MODEL UNIT FOR

IRON ENRICHED CHICKEN MEATBALLS & NUGGETS

Chicken Meat Balls Chicken Nuggets

A) NON-RECURRING EXPENDITURE : (For preparing products with 5 Kg chicken

meat/batch)

1. Land, Land-development and other costs Free, if owned by the

Entrepreneur

2. Building

3. Equipments : The following equipments with approximate price are required

Sl

no

Name Qty(no) Capacity Approx Price

(Rs.)

1 Killing cone with stand 5 cones 5 birds 28000.00

2 Scalding tank 1 5 birds 35000.00

3 De-feathering machine 1 5 birds 12500.00

4 Utility table (Stainless

steel)

1 Small size:

180x84x90cm

13600.00

91

5 Trays (Steel) 4 16 x 32 x 1.25

inch

4000.00

6 Knives (L/S) 4 - 1000.00

7 Disposable drum/ dustbin 2 20-25 litres 2000.00

8 Food grade anticoagulant

(Trisodium citrate

@0.3g/100ml blood)

1 500g 1700.00

9 Blood collection jar

(Plastic)

3 100 ml 240.00

10 Deep freezer cum chiller

(-180C)

1 350 litres 26120.00

11 Mincer (meat) Electric 1 5 kg 4000.00

12 Mould (metal) 4 4x4x8 inch 600.00

13 Momo steamer 1 12 litre 5000.00

14 Cooking vat Aluminium

top

1 29-36 Inch

diameter; approx

30 Kg

5100.00

15 Packaging material

(LDPE)

1 pack 250 g (3 micron

thickness),

200pcs/pack

500.00

16 Sealing machine (for

aerobic packaging)

1 - 2200.00

17 Cooking arrangement

a) Burner 1 - 2400.00

b) LPG gas 1 - 800.00

18 Measuring plastic cups

(Graduated)

1 set 50 to 500ml 250.00

19 Weighing balance (Dial

type- plastic)

1 5 Kg 1500.00

20 Weighing balance (Plastic

Digital)

1 1 kg 1200.00

21 Dial Thermometer

(graduated up to 100°C)

1 - 700.00

Total 148410.00

Thus, total non-recurring expenditure is Rs. 148410.00

92

B) RECURRING EXPENDITURE : (For preparing products with 5 Kg chicken meat/batch)

1. Cost of live spent chicken: Rs. 180/ kg x 8 kg = Rs 1440.00

(Assuming that 8 kg meat yields 5 kg deboned meat after processing)

2. Non-meat ingredients requirement for 5 Kg chicken meat balls preparation:

SL.

No

Non meat

ingredients

Quantity Approx Price

(Rs.) Total (Rs.)

1 Maida 250g 60/Kg 15.00

2 Soya flour 250g 75/Kg 20.00

3 Cooking oil 250ml 120/litre 30.00

4 Condiments

250g

167g

Garlic

150/kg 25.00

83g

Ginger

150/kg 15.00

5 Spices 75 1000/kg 75.00

6 Salt 90 20/kg 2.00

7 Blood 250ml - -

Total 182.00

3. Non-meat ingredients requirement for 5 Kg chicken nugget preparation

SL. No

Non meat

ingredients

Quantity Approx Price

(Rs.) Total (Rs.)

1 Maida 250g 60/Kg 15.00

2 Soya flour 350g 75/Kg 26.00

3 Cooking oil 250ml 120/litre 30.00

4 Condiments

250g

167g

Garlic

150/kg 25.00

83g

Ginger

150/kg 15.00

5 Spices 75 1000/kg 75.00

6 Salt 90 20/kg 2.00

93

7 Blood 550ml - -

Total 188.00

4. Electricity cost (L/S.) : Rs. 2000 per month*(per day Rs.67.00)

5. Labour cost : Rs. 12,000 per month** (per day Rs. 400.00)

Thus, total recurring expenditure to make products with 5 Kg chicken meat per day is

(1440+182+188+67+400) = Rs. 2277.00

*To be calculated based on the number of batches of 5Kg of processed meat product

preparation in a day.

** Based on the Daily labour wage, it may be free if family members are involved in the

activities.

NB:

1. The chicken meat balls and the chicken nuggets thus prepared will have iron contents of

99.62 and 184.49 per cent, respectively higher than those normally available.

2. This model has been prepared focussing on Guwahati using a minimum unit of 5 Kg

deboned meat per batch. The cost of production will continue to decrease as the scale of

operation will increase.

Minced chicken meat Mincing harvested chicken meat

Slaughtered Spent hen harvesting Deboned spent hen meat

Meat Balls

T4 (10% Blood) Meat Balls

T3 (7.5% Blood)

Meat Balls

T1 (0% Blood)

Meat Balls

T2 (5% Blood)

Aerobic & Vacuum Packaged

Meatballs T1 (0% Blood)

Aerobic & Vacuum Packaged

Meatballs T2 (5 % Blood)

Aerobic & Vacuum Packaged

Meatballs T3 (7.5% Blood)

Aerobic & Vacuum Packaged

Meatballs T4(10% Blood)

Nuggets

T1 (0% Blood)

Nuggets

T2 (11% Blood)

Nuggets

T3 (14% Blood)

Nuggets

T4 (17% Blood)

Aerobic & Vacuum Packaged

Nuggets T1 (0% Blood)

Aerobic & Vacuum Packaged

Nuggets T2 (11% Blood)

Aerobic & Vacuum Packaged

Nuggets T3 (14% Blood)

Aerobic & Vacuum Packaged

Nuggets T4 (17% Blood)

Homogenising sample Filtration process for pH

Estimation of Iron Microbial count

Enterprenurship Training programme for Technology Transfer (27-02-2020)

Enterprenurship Training programme for Technology Transfer(07-03-2020)

NABARD Research Series

S. No. Title of Study Agency 1. Whither Graduation of SHG Members? An

exploration in Karnataka and Odisha National Bank for Agriculture and Rural Development (NABARD)

2. Study on Strengthening the value chain of TDF Wadi Projects in Andhra Pradesh

Administrative Staff College of India, Hyderabad

3. Developing a roadmap of Social Enterprise Ecosystem- as a precursor for a viable Social Stock Exchange in India

Grassroots Research and Advocacy Movement (GRAAM)

4. Sustainability of Old Self Help Groups in Telangana

Mahila Abhivrudhi Society, Telangana

5. Impact Assessment of RuPay Card on Weaker and Marginalized Sections in Bihar and Uttar Pradesh

Rambhau Mhalgi Prabodhini, Mumbai

6. Getting More from Less: Story of India’s Shrinking Water Resources

Indian Council for Research on International Economic Relations (ICRIER)

7. Identifying the Most Remunerative Crop-Combinations Regions in Haryana: A Spatial- Temporal Analysis

Centre for Research in Rural and Industrial Development (CRRID)

8. Climate Change Impact, Adaption and mitigation: Gender perspective in Indian Context

ICAR- National Institute of Agricultural Economics and Policy Research (ICAR-NIAP)

9. Achieving Nutritional Security in India: Vision 2030

Indian Council for Research on International Economic Relations (ICRIER)

10. Development of Iron Enriched Spent Hen Meat Products for Boosting Layer Industry and Entrepreneurship

Assam Agriculture University, Guwahati

NATIONAL BANK FOR AGRICULTURE AND RURAL DEVELOPMENT

Plot No. C-24, 'G' Block, Bandra Kurla Complex Rd, Bandra East, Mumbai, Maharashtra 400051