research regarding the physiological adaptation and … · 2013-02-07 · prof. mire Şan vioara...

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UNIVERSITY OF AGRICULTURAL SCIENCES AND VETERINARY MEDICINE CLUJ-NAPOCA

USAMV PhD SCHOOL FACULTY OF ANIMAL SCIENCE AND BIOTECHNOLOGIES

ENG. IULIA EUGENIA FE ŞTIL Ă

RESEARCH REGARDING THE PHYSIOLOGICAL ADAPTATION AND THE INFLUENCE OF SOME FACTORS ON METABOLIC PROFILE IN DAIRY

CATTLE

SUMMARY OF THE PHD THESIS

SCIENTIFIC COORDINATOR: PROF. MIREŞAN VIOARA Ph.D.

CLUJ-NAPOCA 2012

II

SUMMARY

of the PhD Thesis

RESEARCH REGARDING THE PHYSIOLOGICAL ADAPTATION AND THE INFLUENCE OF SOME FACTORS ON METABOLIC PROFILE IN

DAIRY CATTLE

elaborated by Eng. FEŞTILĂ IULIA EUGENIA, under the scientific supervision of Prof. MIREŞAN VIOARA Ph.D., from the University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca

The PhD Thesis “RESEARCH REGARDING THE PHYSIOLOGICAL ADAPTATION

AND THE INFLUENCE OF SOME FACTORS ON METABOLIC PROFILE IN DAIRY

CATTLE” is structured in two parts and contains 11 chapters.

INTRODUCTION

Consumer propensities are generally oriented towards animal protein that is provided

in most mammals and birds exploiting farm products and by-products which have high

biological value. In recent years, in Romania, the population became increasingly aware of

the importance of food quality for the harmonious development of the human body. In this

respect, our country is already perceived the Western influence of a healthier diet. Romania

currently has outstanding opportunities for implementing organic farming practices, both

due to lower level of use of chemical fertilizers in agriculture and the context of increasing

demand for organic products in Western markets.

The study of metabolic profile in dairy cattle, gives us an insight into animal health,

which will be found in the quantity and quality of the products obtained.

By this doctoral thesis we wanted to bring some contribution to the knowledge of

factors that influence the metabolic profile of cattle bred and exploited in different systems

products. Also, with implementing the results, we tried to develop relevant conclusions and

recommendations, with which technologies can be improved increasing the welfare of dairy

cattle and the products obtained.

In our country, growth and exploitation of animals in organic farming has grown,

especially since the EU has implemented the concept of welfare, which was adopted and

implemented from us. Establishment of organic farms for dairy cattle growth requires

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knowledge of physiological factors and their metabolic profile, which is a requirement for

application of appropriate technology use.

Our research fit into the theme of European research on sustainable agriculture, biodiversity

conservation and production of animal products that ensure the bio security of human food.

PART I

BIBLIOGRAPHIC STUDY

CHAPTER I NEWS AND PERSPECTIVES IN GROWTH AND

OPERATION OF CATTLE

This chapter highlights some issues relating to the cattle global and national levels, as

well as factors that led to the rearing of cattle and increased yields obtained from both

conventional and organic systems.

FAO statistics shows the upward trend of yields obtained from cattle in most

countries. Thus, if in 2000, world milk production was 490.611 million tons, in 2009 it

reached 583.401 million tons

Table I.1

Dynamics of total milk production of cows during 1980 – 2009 (th tones)

(Sources: www.fao.org; Buletin FAO 2010)

Anii de referin ţă/ Years of reference Specificare Specification 1980 1991 2000 2001 2003 2005 2007 2009

2009/1980 %

2009/1991 %

Pe glob/world 417200 460993 490611 505746 479537 529662 531563 583401 139,83 115,35 AFRICA 10618 15178 19762 20993 21118 21519 21720 28964 272,78 137,96 Egipt 648 691 1638 1900 1900 2300 2300 3200 493,82 168,42 AMERICA DE NORD 76419 84171 97290 98947 99108 88388 88420 94074 123,10 95,07 Canada 7830 7915 8090 7960 7880 8100 8215 8213 104,89 103,17 Mexic 6949 6336 9311 9658 9842 9898 9912 10549 151,80 109,22 S.U.A. 58139 66423 77023 77248 77252 80287 80350 85859 147,67 111,14 AMERICA DE SUD 24082 31986 44722 46218 46312 48066 48530 60988 253,25 131,97 Argentina 5311 6375 10121 8500 7900 8100 8183 10366 195,17 121,95 Columbia 2187 4017 5600 6020 6055 6770 6832 7545 344,99 125,33 ASIA 33026 55041 95725 102509 104685 122098 124352 150632 456,10 146,94 China 1191 4816 7138 11612 11685 24200 26800 35509 2981,44 305,79 India 17318 27700 34000 35300 36500 38500 39300 45140 260,65 127,87 Japonia 6526 8169 8497 8385 8360 8282 8268 7909 121,92 94,32 Turcia 3732 3000 9000 8620 8638 9500 9520 11583 310,35 134,37 EUROPA 170258 162660 209626 211524 210564 209334 207253 208141 122,25 98,40 Austria 3434 3344 3340 3292 3450 3159 3200 3229 94,03 98,08 Finlanda 3236 2712 2450 2532 2471 2595 2586 2332 72,05 92,42 Franţa 26720 26334 24999 25197 24614 25282 25436 23341 87,35 92,64 Germania 33754 30976 28331 27874 28350 27600 27432 27938 82,76 100,22 Elveţia 3653 3892 3889 3944 3900 3800 3700 4073 111,49 103,27 Italia 10546 10000 12444 11353 11000 10500 10323 12219 115,86 107,62 România 4038 3450 4301 4637 4852 5720 5830 5208 128,97 112,31 U.K 15917 14976 14488 14869 15056 14577 13542 13236 97,74 89,01 Oceania 12240 14126 23486 25555 25065 24717 23720 24855 203,06 97,26

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The main way to increase production efficiency, especially is the quality and

potential of biological material existing in livestock, while biological material has

undergone an intensive program of improvement on production lines, which has now

resulted in improved breeds specialized on production lines.

In recent years, in Romania, the population became increasingly becoming aware of

the importance of quality food for the harmonious development of the human body. In this

respect, our country is already perceived Western influence a healthier diet. Romania

currently has outstanding opportunities for implementing organic farming practices, both

due to lower level of chemical inputs in agriculture and context of increased demand for

organic products on Western markets.

Development of organic farming in Romania in recent years was based on

requirements for organic products, the EU could meet only 27% of import needs-European

investment required and because organic farming in our country. Demand for organic

products is increasing it may be a new opportunity for Romanian agriculture exports.

Table I.2 Situation of organic farming in Romania (thousands)

CHAPTER II

METABOLIC PARTICULARITIES AND FACTORS THAT ARE AFFE CTING

THE METABOLIC PROFILE OF DAIRY CATTLE

Living organisms carry out a continuous exchange of energy and information with

the external environment, they are open systems in thermodynamic terms. Metabolism

conducted under permanent interaction between living and inanimate matter. Due to this

Indicatori / Indicators 2006 2007 2008 2009 2010 2011

Number of registered organic operators 3409 3834 4191 3228 3155 9703

Area cultivated in organic farming, arable 45605 65112 86454 110014,4 148033,5 147581,55

Organic Area, (ha) pastures and meadows 51200 57600 46006,5 39232,8 31579,11 78197,51

Organic, permanent crops (ha) orchards and vine 294 954 1518 1869,4 3093,04 4166,62

Collection of spontaneous flora 38700 58728 81279 88883,4 77294,35 338051

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issue, in Chapter II we conducted a literature study on metabolic particularities of dairy

cattle and factors influencing their metabolic profile. Were described aspects of metabolism

of nutrients (proteins, carbohydrates, fats, minerals, vitamins). The factors that influence the

metabolic profile of dairy cows were: nutrition, reproduction and climatic factors.

CHAPTER III OXIDATIVE STRESS IN DAIRY CATTLE

In recent years assessing oxidative stress contributed to knowledge of fundamental

mechanism involved in metabolic disorders, particularly important for dairy cattle.

Therefore, in Chapter III, we conducted a literature study on the mechanism of oxidative

stress and its monitoring of biomarkers. Were described enzymatic antioxidant defense

systems of the body that are designed to protect cells from excess reactive oxygen species

(ROS) and free radicals and oxidative stress are indicators of tissue.

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PART II PERSONAL RESEARCH

CHAPTER IV

THE ORGANISATION OF EXPERIMENTAL DEVICE AND RESEARC H DEVELOPMENT

4.1 THE AIM AND THE OBJECTIVES OF THE EXPERIMENTS

The aim of the PhD Thesis

In the present research and experiments from the Thesis, the aim was to highlight

"PHYSIOLOGICAL ADAPTATION AND INFLUENCE OF SOME FACTORS ON

METABOLIC PROFILE IN DAIRY COWS."

The objectives of this thesis:

1. Organizing experimental model in two different operating systems and growth:

conventional and ecological systems;

2. Monitoring the studied biological material, on physiological states, the production levels,

and the seasons and different metabolic profiles in two farms;

3. Quantitative and qualitative determination of milk by lactation according to physiological

and reproductive parameters;

4. Determination of the phenotypic correlation between production performance and milk

production traits at the biological material studied;

5. The influence of season on the main quality indices of milk;

6. Chromatographic determination method of bovine milk retinol;

7. The determination of the authenticity of products from organic milk and determining their

quality parameters;

8. Determining major polymorphisms of milk in order to make an early marker assisted selection;

9. Research on the influence of physiological state on metabolic profile and hematology

parameters in dairy cattle;

10. Research on the influence of seasons on metabolic profile and hematological parameters

in dairy cattle;

11. Determining the oxidative stress status of biological material studied by using specific

markers;

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4.2 THE ORGANISATION OF THE EXPERIMENTS

The experiments were conducted during the years 2010-2012, in two farms with

different operating conditions and growth and different dairy breeds:

- farm SC Agrolact SRL Aiud, Alba County (denoted farm "A"), where its grown and

exploited in conventional system a population of dairy cattle from Bălţată Românească

Simmental breed.

- farm A.F. Mihalca V. Ioana-Petrova, Maramureş County (denoted farm "B"), where its

grown and exploited in organic system a population of dairy cattle from Brună of

Maramureş breed. We mention that the farm is organic certified from 2007.

4.3 THE BIOLOGICAL MATERIAL

The biological material studied was the population of dairy cattle Bălţată

Românească Simmental and Brună of Maramures breeds. The biological material studied

was represented by 267 heads of dairy cattle from farm "A" that we have plots based on

lactation and 50 heads of dairy cows on the farm "B" also plots according to lactation.

Of all biological material studied three groups were formed in both dairy breeds were

followed from lactation I to lactation III:

Biological material

Specification

UM Farm „A”

Bălţată Românească Simmental

Farm „B”

Brună de Maramureş

Lactation I - Lot I Heads 17 13

Lactation II - Lot II Heads 15 16

Lactation III - Lot III Heads 18 13

Total Heads 50 42

The biological material which has been the object of our experiments was operated

under different management (conventional and organic) and the feed was a unique mixture

being made in each firm from its own resources. Feeding was done "ad libitum" with unique

feed both summer and winter.

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4.4 METHODS

In the experiments for this thesis were used the following methods:

� to determine the production performance of the biological material in the study, data

were taken from farms, each with a base of information on biological material;

� were followed the first calving age, total and normal lactation, total milk production,

milk normal production, calving interval, breast rest, the dynamic completed

lactations;

� was monitored quantity and quality of milk and breeding indices;

� was followed the authenticity of organic products;

� the determination of hematological parameters were followed by automatic analyzers

Abacus Junior Vet and Vet Myndray BC, which utilize whole blood analyzers;

� blood biochemical indices were determined using veterinary apparatus Abaxis

VetScan VS-2 with rotors for the bovine species;

� antioxidant enzymes GPx and SOD were determined by UV-Vis spectrophotometer

Master Touch Screen;

� to determine the qualitative characteristics of milk were used MilkoScan devices,

Ecomilk (protein, fat, lactose, SUN), Fossomatic (number of somatic cells) and

BactoScan (total number of germs);

� milk retinol determination was performed by HPLC system with Waters software for

data processing;

� to identify polymorphic profile of milk proteins was performed by is electric

focusing technique (IEF);

� results were statistically processed using Microsoft Excel software, Graph Pad

ANOVA and IBM SPSS;

All these modern methods that were used, and measurements were performed in

research laboratories accredited from USAMV Cluj-Napoca.

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4.5 THE ORIGINAL ELEMENTS OF PHD THESIS

In this thesis, we consider the elements of originality, the following:

• the choice of experimental model and organizational procedures;

• the study of the metabolic profile and authenticity of the organic farm;

• Study on the influence of seasons on hematological profile and blood

biochemical indices in the two breeds studied;

• assessing oxidative stress through antioxidant enzymes glutathione peroxides

and superoxide dismutase based on physiological status;

• Provitamin A determination of organic milk compared with those obtained

from conventionally farmed cattle;

• establishing correlations between the type of polymorphism and metabolic

profile in order to improve early in the study of biological material;

• Study major polymorphisms of milk in order to make an early marker assisted

selection

CHAPTER V THE PRODUCTIVE PERFORMANCE OF THE STUDIED BREEDS

Trough the specific aptitudes, the race affects most quantitative and qualitative

characteristics of milk, and many other important features that contribute to the achievement

of milk production, such as precocity, longevity, feed conversion ability. In order to

characterize the main indices of milk production for cattle farms populations studied, as

indicators of milk production were considered total and normal lactation, total milk

production, fat and protein, normal production milk, fat and protein. They revealed to

establish the main quantitative and qualitative milk production. At the biological material

studied were highlighted also the main indices of breeding.

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5.1 THE AIM AND OBJECTIVES OF THE EXPERIMENT

The aim of our research was to highlight the production performance of two

populations of dairy husbandry systems operated. Thus, we sought production performance

throughout the experimental farm with dairy Bălţată Românească Simmental breed

operating in conventional farming system and the performance of the population Brună of

Maramures breed, organically farmed.

5.2 RESULTS AND DISCUSSIONS

The biological material studied namely the 50 heads of dairy cattle on three

lactations effectively prosecuted total existing farm "A", is characterized by an average

production potential of 5620, 96 kg of milk per lactation normal and 6479.56 kg on total

lactation milk.

In terms of quality, milk fat content had an average around 3.9% on total lactation

and normal lactation, with a nearly linear trend an the differences were small and

insignificant. Average content of protein substances, was 3.3% and 3.29% on total lactation

and normal lactation. The total amount of fat on lactation stood around 252.94 kg, 221.18 kg

on total lactation ,respectively on normal lactation. The total amount of protein had an

average of 214.12 kg and 185.31 kg on total lactation and normal lactation.

As regards biological material studied, the population of dairy cattle from Brună of

Maramures breed, he showed the average of quantities of milk production per lactation

3274.38 kg 3223.54 kg total and normal lactation milk.

Milk quality is enhanced by an average of 3.92% fat and 4.00% on total lactation

and normal lactation and a protein content of 3.52% and 3.53% on total lactation and normal

lactation. Milk fat content and protein has an evolutionary character almost linearly, the

average total lactation fat being 128.35 kg and 129.26 kg on total and normal lactation

while the amount of protein was 115.84 kg per lactation 113.06 kg on total and normal

lactation.

Knowing that the correlation characteristics are particularly important for accurate

and objective selection method. To this end, we calculated correlations between milk

production traits of biological material studied in both populations of dairy cows . Positive

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correlations based on intense obtained in our experiment, between the amount of fat and the

amount of milk fat content and the amount of protein in most lactation analyzed and studied

both races, we conclude that selection applied after the fat milk would improve correlated

with other features thereof.

CHAPTER VI

RESEARCH REGARDING THE INFLUENCE OF THE SEASON ON T HE MAIN MILK QUALITY PARAMETERS IN B ĂLŢATĂ ROMÂNEASCĂ SIMMENTAL

AND BRUNĂ OF MARAMURE Ş CATTLE POPULATIONS


The aim of our research was to highlight the influence of season on quality

parameters of milk from cows raised in a conventional farm system "A" and organically

farmed dairy farm "B".

Milk quality assessment was done by determining the following parameters: grease

G, P protein, lactose, non-fat dry substance SUN, somatic cell count SCC, the total number

of germs NTG and retinol in all four seasons.


Effects of seasons, should be considered as important factors with influence on milk

quality parameters. Seasons of the year are related more with different diets. They depend

on the nature and quality of feed required to provide the animal physiological needs,

changes in milk composition were more related to feeding factors than genetics.

Following the results we see that the qualitative features of milk showed variations

depending on the production season and growing conditions (Tables 26 and 29).

Fat content was influenced by specific race and food, and our experiments shown that the

lowest percentage of fat was recorded in summer in both populations studied, 3.81 ± 0.03%

for the population of Bălţată Românească and 3.44 ± 0.03% for Brună breed population and

the highest percentage in the winter of 4.07 ± 0.07% and 4.13 ± 0.13%

Protein content showed values between 3.18 ± 0.01% in summer and 3.32 ± 0.04% in winter

at the population from Bălţată Românească breed and 3.26 ± 0.05% during summer and

XII

3.63 ± 0.08% during Winter at the population from Brună breed, a low protein levels due to

the change in diet.. The low content of lactose in Summer season of 4.49 ± 0.08% for the

population of Bălţată Românească breed and 4.55 ± 0.05% and for the population of Brună

breed, are a consequence of decreased synthesis of lactose in the mammary gland, which

occurs in summer. SUN non-fat dry matter was influenced by the proportion of proteins,

lactose and minerals, values and ranged from 8.65 ± 0.04% 8.86 ± 0.07% in winter and

autumn, for Bălţată Românească breed population and between 8.4% and 9.04% autumn

season in the winter for Brună breed population.

The content of retinol (vitamin A) presented in summer the highest values in both

populations, of 47.09 ± 1.12 ug/100 ml at Bălţată Românească and 50.05 ± 1.31 ug/100 ml

at Brună, values that were influenced by diet rich in green mass which is a rich source of

carotenoids.

Fig. VI.1 HPLC Chromatogram of retinol separation from milk at a cattle population from Bălţată Românească Simmental breed

NTG and SCC values were within the limits imposed by national regulations

concerning quality and hygiene of cow's milk for processing (NTG less than or equal to

100.000/ml and SCN less than or equal to 400.000/ml). The highest values of NTG and

SCC were recorded in summer in both populations of 74.45 ± 4.51 thousand / ml and 171.35

± 28,700 / ml at Bălţată Românească breed and 96.4 ± 12.03 thousand / ml and 142 ± 24

020 / ml for Brună of Maramures breed.

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 min

-2

-1

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

mAU350nm,4nm (1.00)

0.77

4

2.50

9

2.68

2

2.90

1

3.29

8

XIII

CHAPTER VII RESEARCH REGARDING THE AUTENTICITY OF ORGANIC PRODU CTS

OBTAINED FROM DAIRY CATTLE MILK


The aim of our research was to highlight the main indices of quality of finished

products from organic milk produced on the farm AF Mihalca V. Ioana Petrova.

To highlight the quality of the finished product were pursued: physic-chemical parameters

(water content, fat percentage, protein percentage) and microbiological parameters

(Escherichia coli, coagulate positive staphylococci and coli forms.)


Cheese samples taken from the three types were consistent in terms of the STAS

effect on color: white with yellowish tint without foreign materials with flavor cheese,

specific to each type of cheese.

Following the results obtained in range " Telemea ecologică de Petrova " have shown

an average moisture percentage of 46.45%, an average percentage of fat in dry matter of

40.70% and an average of 3 NaCl, 33%. Results for "Caşcaval ecologic de Petrova" found a

41% percent moisture, 82%, the average percentage of fat in dry matter 44.70% and 2.92%

salt. Assortment of "Brânză frământată ecologică de Petrova" has revealed qualitatively by a

percentage of 42.86% water moisture, percentage of fat in dry matter 45.79% and a rate of

2.62% salt.

In the cases of the three types of cheese was found that E. coli was absent. Coli form

bacteria in cheese mixed showed an average of 7 ± 3.24 / g, and cheese 5 ± 3.16 / g For

Staphylococcus coagulate positive, the values were <100.

Following research on quality parameters of products from organic milk, it appears that

they meet current standards both physic-chemical and microbiological making organic cheese

sortimentele Petrova be appreciated and sought after by consumers (tables 32 and 33).

XIV

CHAPTER VIII THE SUDY OF MAJOR MILK PROTEIN POLYMORPHISMS IN TWO CATTLE

POPULATIONS BELONGING TO B ĂLŢATĂ ROMÂNEASCĂ SIMMENTAL AND BRUNĂ OF MARAMURE Ş BREEDS


The aim of our research was to study the major milk protein polymorphism in two

populations of cattle in experiment.

The primary objective was to determine the polymorphic profile of the six major

milk proteins, to determine the genetic structure of two populations of dairy cattle from

Bălţată Românească Simmental and Brună of Maramureş breeds


The analysis of electrophoresis profiles revealed at the 6 loci encoding the 6 major

types of milk proteins presence of common alleles in both populations of dairy cows (Table

VIII.1). It was found that the two breeds studied were found different frequencies of alleles

in 4 of the 6 loci studied. The frequency of alleles associated in various studies with higher

quality milk and cheese obtaining higher yields (αS1-CN C, β-CN A2, B β-CN, K-CN B, β-

LG B) is higher at Brună breed compared to Bălţată Românească breed. This is explained

by the fact that the Brună Marmureş breed breed was formed from Brună Schwyz, breed

that has been improved in increasing milk quality, which is intended mainly for processing.

Bălţată Românească breed was formed from Simmental and Sură de Stepă cattle, improved

breed towards producing a higher quantity of milk. This explains the relatively higher

frequency of alleles that have been associated in various studies with a larger quantity of

milk (B αS1-CN, β-CN A1, β-LG A) compared to those observed in the Brună breed.

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Tabel/Table VIII.1 Genetic structure in αS1-CN locus ,β-CN, αS2-CN, K-CN, α-LA, locusul β-LG la at the cattle population from Bălţată Românească Simmental şi Brună de Maramureş breed

CHAPTER IX

RESEARCH REGARDING THE INFLUENCE OF THE PHYSIOLOGIC AL STATE ON METABOLIC PROFILE AND HAEMATOLOGICAL INDIC ES ON

DAIRY CATTLE


The aim of our research was to highlight the biochemical and hematological

parameters of the biological material studied and its changes depending on the physiological

state. We followed the metabolic profile and hematological parameters in the last month of

gestation and the first month of lactation, which was divided in early lactation (0-7 days)

and advanced lactation (25-35 days). For metabolic profiling were determined 11

parameters: aspartate aminotransferase (AST), gamaglutamiltransferaza (GGT), alanine

aminotransferase (ALT), calcium (CA), magnesium (Mg), phosphorus (P), urea, cholesterol,

Bălţată Românească Simmental Brună de Maramureş Locusul Genotiypes Nr.

Indivizi/ genotipuri

Frecvenţa genotipurilor

Frecvenţa de genă

Nr. Indivizi/

genotipuri

Frecvenţa genotipurilor

Frecvenţa de genă

BB 30 0.75 17 0.68 BC 7 0.17 5 0.20

αS1-CN

CC 3 0.08

pB=0.84 qc=0.16 3 0.12

pB=0.78 qc=0.22

A1A1 7 0.17 2 0.08 A1A2 16 0.40 3 0.12

A2A2 11 0.28 14 0.56

A1B 3 0.07 3 0.12

A1C 1 0.03 - -

β-CN

A2B 2 0.05

pA1=0.43 qA2=0.50 mB=0.06 nC=0.01

3 0.12

pA1=0.20 qA2=0.68 mB=0.12

AA 13 0.32 4 0.16 AB 23 0.58 10 0.40

K-CN

BB 4 0.10

pA=0.61 qB=0.39 11 0.44

pA=0.36 qB=0.64

α-LA BB 40 1 pB=1 25 1 pB=1 AA 9 0.22 5 0.20 AB 25 0.63 12 0.48

β-LG

BB 6 0.15

pA=0.54 qB=0.46 8 0.32

pA=0.54 qB=0.46

XVI

total protein (PT) , albumin (ALB) and gamma globulin (γGLOB). Blood picture was

composed of 7 parameters: the number of white blood cells (WBC), erythrocyte count

(RBC), hemoglobin (Hb), hematocrit (Ht), mean corpuscular volume (MCV), mean red cell

hemoglobin (MCH) concentration mean erythrocyte hemoglobin (MCHC).


Our experiment results showed that early gestation and lactation are physiological

states leading to significant changes in hematological variables (Tables 54 and 55).

Variations on leukocyte counts showed a growth in advanced gestation in both populations

of dairy cows compared with early lactation period when they recorded the lowest values,

values in advanced gestation being 11.62 ± 0.26 X103 / mm3, the Bălţată Românească

Simmental breed population respectively 11.28 ± 0.28 X103 / mm3 for the people of Brună

Maramures breed, showing the total number of leukocytes increases only after 10 days after

parturition.

RBC levels increased in advanced pregnancy because of increased fetal growth during

this period that requires a greater amount of oxygen, the values obtained are 5.99 ± 0.07

x106/mm3 at the population from Bălţată Românească Simmental breed and 6.62 ± 0.07

x106/mm3 at the population from Brună Maramures breed

Low hemoglobin Hb during pregnancy advanced, compared with lactating advanced,

is because the female body in advanced gestation mobilizes hemoglobin in the fetal

circulation and the fact that the average life blood erythrocytes from pregnant females

compared with those during lactation is reduced.

Ht hematocrit value increased during pregnancy advanced in both populations of dairy

cows is determined by the increase in volume of erythrocytes (RBC), which in turn

increased the volume of water during pregnancy, specifying Ht during this period was of

27.31 ± 0.29% to ± 0.34 at Bălţată Românească and 31.4% at Brună of Maramureş. Level

erythrocyte indices MCV, MCH and call was influenced in both breeds Hb hemoglobin and

hematocrit Ht, there is a link between MCV mean corpuscular volume and mean erythrocyte

hemoglobin HEM, meaning that an increase in MCV in early lactation resulted increased

HEM same physiological and decreased MCV in advanced pregnancy entailed and

subtracting HEM.

XVII

Following the results obtained, it is observed that the values of the 11 blood

biochemical parameters track falls within the specialty literature cited, and the differences

are due to metabolic changes of animal body undergoes during pregnancy and lactation

(Tables IX.1 and IX.2).

Table IX.1

Mean values and variability of blood biochemical indices according to physiological state at the cattle population from Bălţată Românească –Simmental breed

ALAT- Alanine aminotransferase; Ca-Calcium; Mg -magnesium, P-Phosphorus; BUN-blood ureea nitrogen; Cholesterol; TP- Total protein; ALB -Albumin; γGlob- gammaglobulina; * Smith, 2009; ** University Davis of California , 2011; *** Radostis, 2007;

Table IX.2

Mean values and variability of blood biochemical indices according to physiological state at the cattle population from Brună de Maramureş breed


Increased activity of aspartaminotransferase AST and GGT gamaglutamiltransferase

during pregnancy and lactation advanced early (0-7 days) in both populations studied, is

because during pregnancy, aminotransferase enzymes have a role in the survival and

Late gestation Early lactation (0-7 days) Lactation (25-35 days) Variables

Specification

UM

n

Limits

X ± sx V% X ± sx V% X ± sx V% ASAT U/L 50 43-127* 53.58 ± 0.36 4.79 57.11 ± 0.43 5.53 49.06 ± 0.76 10.98 GGT U/L 50 15-38* 29.56 ± 0.70 16.87 34.62 ± 0.68 13.89 28.94 ± 0.36 8.81

ALAT U/L 50 11-40** 22.06 ± 0.28 9.17 20.14± 0.54 19.16 24 ± 0.30 9.09 Ca mg/dl 50 8.5-11.6** 9.39 ± 0.13 10.09 8.41 ± 0.07 6.10 9.12 ± 0.15 12.11 Mg mg/dl 50 1.7-2.87 *** 2.81 ± 0.06 15.10 2.39 ± 0.03 10.34 2.66 ± 0.02 7.33 P mg/dl 50 4.95-6.8** 5.28 ± 0.01 2.51 6.08 ± 0.02 2.34 5.80 ± 0.05 6.14

Uree mg/dl 50 6-27*** 23.16 ± 0.32 9.85 20.42 ± 0.17 6 24.96 ± 0.38 10.96 Colesterol mg/dl 50 65-220 *** 111.06 ±1.14 7.26 115.3 ± 0.62 3.38 134.7 ± 3.78 19.89

PT g/dl 50 5.7-8.1 *** 6.80 ± 0.08 8.99 6.27 ± 0.04 5.56 6.4 ± 0.04 5.30 ALB g/dl 50 2.1-4 *** 2.71 ± 0.01 5.09 2.13 ± 0.03 10.87 2.61 ± 0.01 5.09 γGlob g/dl 50 1.7-3.4** 1.94 ± 0.04 15.24 2.31 ± 0.03 11.91 1.92 ± 0.01 5.98

Late gestation Early lactation (0-7 days) Lactation (25-35 days) Variabile/ Variables

Specification

UM

n

Limits

X ± sx V% X ± sx V% X ± sx V% ASAT U/L 42 43-127* 48.86 ± 0.45 5.99 52.73 ± 0.63 7.71 46.6 ± 0.52 7.33 GGT U/L 42 15-38* 24.84 ± 0.30 7.93 28.05 ± 0.28 6.47 22.55 ± 0.27 7.76

ALAT U/L 42 11-40** 24.5 ± 0.40 10.69 23.15 ± 0.48 13.44 26.7 ± 0.36 8.80 Ca mg/dl 42 8.5-11.6** 10.05 ± 0.05 3.28 9.17 ± 0.04 3.38 9.64 ± 0.05 3.52 Mg mg/dl 42 1.7-2.87*** 2.8 ± 0.02 5.16 2.37 ± 0.01 5.35 2.71 ± 0.03 5.90 P mg/dl 42 4.95-6.8** 5.84 ± 0.05 5.72 6.11 ± 0.06 7.19 5.54 ± 0.02 5.77

Uree mg/dl 42 6-27*** 24.95 ± 0.24 6.36 22.35 ± 0.34 10.06 28.6 ± 0.29 6.62 Colesterol mg/dl 42 65-220 *** 125.15 ± 1.27 6.28 135.75 ± 0.80 3.86 185.7 ± 0.90 3.16

PT g/dl 42 5.7-8.1 *** 7.20 ± 0.04 4.30 6.2 ± 0.01 1.29 6.60 ± 0.05 5.45 ALB g/dl 42 2-4 *** 4.05 ± 0.03 5.67 2.9 ± 0.03 8.89 3.26 ± 0.02 4.98 γGlob g/dl 42 1.7-3.4** 2.27 ± 0.01 3.96 2.51 ±0.02 7.17 2.12 ± 0.01 3.30

XVIII

development of the embryo and fetus and in early lactation increased serum AST and GGT

level is a consequence of the stress of parturition and initiation of lactation.

Increased calcium in the second period of lactation in both populations at the value of

9.12 ± 0.15 mg / dl at Bălţată Românească Simmental and 9.64 ± 0.05 mg / dl at Brună of

Maramureş represent a physiological mechanism which is achieved by enhancing the action

of vitamin D calcium absorption and bone calcium mobilization by the action of parathyroid

hormone.

Serum total protein and albumin PT WHITE, showed a decrease in both early lactating

dairy cattle populations, because the protein body reserves are mobilized to provide amino

acids needed milk protein synthesis.

Elevated levels of serum globulins in early lactating γGLOB of 2.31 ± 0.03 mg / dL at

Bălţată Românească and 2.51 ± 0.02 mg / dL at Brună of Maramureş breeds, is

demonstrating that is provide the transmission of the passive immunity to newborn .

Low serum urea concentration in both breed during early lactation is own to high

amount of protein needed by the mammary gland for milk production. Reduced dry matter

intake during this period resulted in decreased absorption of ammonia, which resulted in

changes in urea metabolism in the rumen.

Increase lactating cholesterol in the range of 25-35 days in both populations, the

values of 134.7 ± 3.78 mg / dl to 185.7 ± BRS and 0.90 mg / dl in BM is due to increasing

lipid catabolism that occurs with advancing lactation.

Following the results, it was found that the values of hematological and biochemical

blood parameters followed of breeds studied range within the limits cited in the literature,

and these variations are influenced by physiological state.

CHAPTER X

RESEARCH REGARDING THE SEASON INFLUENCE ON METABOLI C PROFILE AND HEMATOLOGICAL INDICES ON DAIRY CATTLE


The aim of our research was to highlight the biochemical and hematological

parameters at the biological material studied and their seasonal changes depending on

climatic factors.

XIX

For the metabolic profilie were determined 10 parameters: aspartate aminotransferase

(AST), gamaglutamiltransferase (GGT), alanine aminotransferase (ALT), calcium (CA),

magnesium (Mg), phosphorus (P), urea, cholesterol, total protein (PT) and albumin (ALB).

Blood hematological profile was composed of 7 parameters: the number of white blood

cells (WBC), erythrocyte count (RBC), hemoglobin (Hb), hematocrit (Ht), mean

corpuscular volume (MCV), mean red cell hemoglobin (MCH) concentration mean

erythrocyte hemoglobin (MCHC).


Following the results obtained from the data presented in Tables 62 and 66 it is

observed that the values of hematological parameters followed of breeds under study, falls

within the specialty literature cited and differences between the four seasons are adaptations

animal body conditions specific environmental and management of the two farming systems

and the values obtained are not covered by pathology.

The high WBC recorded in summer in both populations studied is explained by

increased secretion of cortisol and thus stress due to excessive development of pathogen

affecting animal body and increasing blood viscosity, which may cause allergic effects,

inducing production of WBC. Decrease in RBC during the summer season, at the two

populations studied, is associated with increased in water consumption and reducing the

intake of oxygen, which is the physiological response of the body to be able to maintain

thermal equilibrium under conditions of high temperature . Also decrease heat stress ACTH

levels, which in turn acts by reducing the number of RBC Hb hemoglobin level, depending

on the number of erythrocytes, decreased in both breeds with increasing temperature and

low blood cell count. So, at Bălţată Românească breed Hb level was between 9.29 g / dl in

summer and 9.84 g / dl in the spring season, while at the Brună of Maramures breed, Hb

level ranged from 9, 56 g / dl in summer and 9.82 g / dl in winter. Ht values showed a

minimum in summer when temperatures were high and maximum during spring in both

populations of dairy cows. Explanation of this phenomenon is the increased water

consumption which automatically leads to an increase in volume of erythrocytes, along with

the emergence of hem dilution, case cited by other authors. MCV mean corpuscular volume

XX

level (45.63 to 49.45 fl - BRS, 45.52 to 46.21 fl BM, HEM mean erythrocyte hemoglobin

(16.58 to 17.79 pg - BRS; 15.45 -15.90 pg - BM) and mean erythrocyte hemoglobin

concentration MCHC (35.47 to 37.37 g/100 ml breed BRS, 33.72 to 34.97 g/100 ml breed

BM) were affected in both populations by the seasonal values of hemoglobin and hematocrit

Ht Hb, but were within the limits quoted by literature.

Following the results obtained, it is observed that the values of blood biochemical

parameters 11 track falls within the specialty literature cited (tablesX.1and X.2).

Table X.1 Mean values and variability of blood biochemical indices according to season at the cattle

population from Bălţată Românească –Simmental breed


Table X.2

Mean values and variability of blood biochemical indices according to season at Bălţată Brună de Maramureş breed


Increased serum levels of AST, GGT and ALT in the summer season is an adaptation

of the animal organism at high temperatures, because higher temperatures increase the

reaction rate of physiological processes.

Spring Summer Autumn Winter Variables Specificatio

UM

n

Limits X ± sx V% X ± sx V% X ± sx V% X ± sx V%

ASAT U/L 50 43-127* 48.43 ± 0.58 8.49 51.36 ± 0.31 4.30 48.50 ± 0.59 8.67 47.7 ± 0.38 5.67 GGT U/L 50 15-38* 28.79 ± 0.27 6.12 30.07 ± 0.20 4.84 28.87 ± 0.24 5.94 28.02 ± 0.18 4.59

ALAT U/L 50 11-40** 23.51 ± 0.27 8.16 24.67 ± 0.29 8.59 23.64 ± 0.22 6.80 23.41 ± 0.22 6.80 Ca mg/dl 50 8.5-11.6** 9.24 ± 0.08 6.55 9.40 ± 0.05 4.35 9.21 ± 0.04 3.48 9.14 ± 0.095 4.16 Mg mg/dl 50 1.7-2.87*** 2.33 ± 0.02 6.33 2.39 ± 0.02 7.18 2.29 ± 0.01 5.87 2.45 ± 0.01 4.32 P mg/dl 50 4.95-6.8** 6.21 ± 0.11 13.4 5.97 ± 0.09 10.8 6.26 ± 0.02 3.21 6.47 ± 0.02 2.78

Uree mg/dl 50 6-27*** 20.88 ± 0.20 7 22.36 ± 0.12 3.93 21.38 ± 0.26 8.7 20.38 ± 0.11 3.92 Colesterol mg/dl 50 65-220 *** 112.54 ± 0.60 3.83 109.24 ± 0.74 4.85 113.75 ± 0.87 5.41 116.46 ± 0.38 2.32

PT g/dl 50 5.7-8.1 *** 6.93 ± 0.09 9.62 7 ± 0.03 3.73 6.9 ± 0.05 5.58 6.76 ± 0.03 4.02 ALB g/dl 50 2-4 *** 2.62 ± 0.04 10.9 2.91 ± 0.02 5.88 2.73 ± 0.02 6.45 2.53 ± 0.03 9.04

Primăvara Spring

Vara Summer

Toamna Autumn

Iarna Winter

Variabile/ Variables

Specificaţie

Specification

UM

n

Limite fiziologice X ± sx V% X ± sx V% X ± sx V% X ± sx V%

ASAT U/L 42 43-127* 46.27 ± 0.26 3.73 49.75 ± 0.23 3.11 46.12 ± 0.35 4.92 45.27 ± 0.37 5.32 GGT U/L 42 15-38* 23.25 ± 0.31 8.77 24.12 ± 0.25 6.84 23.55 ± 0.46 12.86 22.91 ± 0.37 10.5

ALAT U/L 42 11-40** 24.28 ± 0.23 6.34 25.17± 0.20 7.70 24.69 ± 0.29 7.73 23.23 ± 0.44 12.4 Ca mg/dl 42 8.5-11.6** 9.68 ± 0.07 5.06 9.99± 0.07 5.10 9.62 ± 0.11 7.79 9.19 ± 0.06 4.78 Mg mg/dl 42 1.7-2.87*** 2.63 ± 0.02 6.84 2.61 ± 0.03 8.42 2.68 ± 0.02 5.90 2.71 ± 0.02 4.79 P mg/dl 42 4.95-6.8** 5.86 ± 0.01 1.87 5.82 ± 0.07 8.07 5.90 ± 0.02 3.22 6 ± 0.04 5.16

Uree mg/dl 42 6-27*** 21.22 ± 0.20 6.26 22.18 ± 0.17 5.04 21.65 ± 0.21 6.51 20.91 ± 0.12 4.01

Colesterol mg/dl 42 65-220 *** 110.02 ± 0.91 5.40 108.53± 0.73 4.37 111.86 ± 0.98 5.72 112.95 ± 0.25 1.45 PT g/dl 42 5.7-8.1 *** 6.9 ± 0.02 2.75 6.95 ± 0.04 3.88 6.74 ± 0.05 5.04 6.47 ± 0.07 7.10

ALB g/dl 42 2-3.6 *** 2.44 ± 0.04 12.29 2.92 ± 0.05 12.8 2.42 ± 0.04 11.98 2.28 ± 0.04 14.0

XXI

The results obtained in our experiment showed an increase in serum calcium in

summer, due to the fact that in the summer diet of the animals was introduced hay, which is

a rich source of minerals, especially Ca. Regarding the magnesium Mg, its absorption in the

rumen decreased due to higher intake of calcium and lower phosphorus. Phosphorus

concentration decrease during summer, recorded in both populations is attributed to

increased consumption of vegetables rich in oil, which contain phytic acid which prevents

complete absorption of phosphorus P.

Increased serum proteins in summer, was a consequence of extracellular fluid loss

due to high ambient temperature and introduction of leguminous plants in the diet, leading

to an increase in total serum protein.

High levels of cholesterol in the cold season, was a consequence of increased

secretion of thyroid hormones, which increase basal metabolism so that animals can ensure

thermoregulation in low temperature conditions.

CHAPTER XI

RESEARCH REGARDING THE EVALUATION OF OXIDATIVE STRE SS ACCORDING TO PHYSIOLOGICAL STATE AT DAIRY CATTLE


The aim of our research was to track whether oxidative stress and antioxidant it self

is involved in the transition from advanced gestation to early lactation and lactation period.

Evaluation of oxidative stress and antioxidant status were made by determining the

antioxidant enzymes glutathione peroxidase GPx and SOD superoxide dismutase.


Low levels of antioxidant enzymes obtained in our experiment in early lactation

(Tables 70 and 72) confirms that oxidative stress increases during the postpartum period.

Increased oxidative stress continuously increases lipid peroxidation products and decreased

levels of antioxidants as metabolic requirements associated with parturition and lactation

periods early lead to higher levels of reactive oxygen species (ROS).

XXII

Antioxidant enzyme GPx levels decreased immediately after parturition and recorded

values of 73.29 ± 2.60 U/gHb at Bălţată Românească breed (Table XI.1) and 71.3 ± 2.06 U /

gHb at Brună of Maramures breed (Table XI.2) because immediately after calving the

increased reactive oxygen species caused a decrease in antioxidant enzyme capacity.

Table XI.1

Mean levels and variability of GPx and SOD according to physiological state at Romanian spotted breed

GPx- glutationperoxidase; SOD- superoxid dismutase

Table XI.2

Mean levels and variability of GPx and SOD according to physiological state at Maramureş brown breed

GPx- glutationperoxidase; SOD- superoxid dismutase

Low levels of antioxidant enzymes obtained in our experiment in early lactation

confirms that oxidative stress increases during the postpartum period. Increased oxidative

stress has continuously increased lipid peroxidation products and decreased antioxidant

levels as metabolic requirements associated with parturition and lactation periods early lead

to higher levels of reactive oxygen species (ROS).

Parameters GPx SOD

Variables

Specification

UM

n

X ± sx V% X ± sx V%

Late gestation

U/g Hb

50

81.3 ± 1.86

16.18

1878.61 ± 48.49

18.25

Early lactation 0-7 days

U/g Hb

50

73.29 ± 1.42

13.76

1786.52 ± 41.82

16.55

Lactation 25-35 days

U/g Hb

50

79.21 ± 2.15

19.22

1828.52 ±59.12

22.86

Parameters GPx SOD

Variables

Specification

UM

n

X ± sx V% X ± sx V%

Late gestation

U/g Hb

42

76.13 ± 0.06

7.53

1756.33 ± 18.48

6.82

Early lactation 0-7 days

U/g Hb

42

71.3 ± 1.31

11.92

1646.96 ± 29.25

11.51

Lactation 25-35 days

U/g Hb

42

75.66 ± 1.03

8.90

1748.66 ± 23.01

8.53

XXIII

CONCLUSIONS

Our research showed that the values obtained of metabolic and hematological profile

in both populations studied range within the limits cited in the literature. Influence of season

on them leads to variations in the values obtained, but these variations are animal organism

adaptation to environmental conditions, ensuring dynamic equilibrium.

Physiological status and influence factors on metabolic profile in dairy cows is very

important to know, because some of these disturbances may damage the health of the cows,

adversely affecting production and reproduction, and finally return units.

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XXIV

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research regarding the physiological adaptation and … · 2013-02-07 · prof. mire Şan vioara...

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