i

FACULTY OF NATURAL SCIENCES

DEPARTMENT OF BIOLOGICAL SCIENCES

KOGI STATE UNIVERSITY, AYINGBA, KOGI SATE

A TECHHNICAL REPORT ON STUDENT INDUSTRIAL WORK EXPERIENCE

SCHEME (SIWES)

CARRIED OUT AT:

DEPARTMENT OF PHARMACOLOGY AND TOXICOLOGY

NATIONAL INSTITUTE FOR PHARMACEUTICAL RESEARCH AND

DEVELOPMENT (NIPRD)

IDU INDUSTRIAL LAY-OUT, IDU, ABUJA

BY:

ODEH ABIGAIL MAIRO

12BS1045

A PRE-REQUISITE FOR THE AWARD OF A GRADE IN SIWES SUBMITTED IN

PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE DEGREE OF

BACHELOR OF SCIENCE IN BIOLOGICAL SCIENCES

January

i

Certification

I Odeh Abigail Mario hereby certify that I wrote the students Industrial Work Experience

Scheme (SIWES) report. The SIWES training to place in the Department of Pharmacology and

Toxicology of the National Institute for Pharmaceutical Research and Development (NIPRD)

Idu, Abuja.

ii

Dedication

I dedicate this report to God Almighty for seeing me through the period of my SIWES

attachment and also to my parents/guardian for their unending support towards building a better

me. I pray that the good Lord reward them.

iii

Declaration

I Odeh Abigail Mario declare that the information in this technical report is my six months

industrial training experience carried out in department of pharmacology and Toxicology,

National Institute for Pharmaceutical Research and Development, Idu Industrial Layout Abuja.

iv

Acknowledgement

I wish to express my profound gratitude to my industry based SIWES supervisor Pham. Lucy

John-Africa a staff in Pharmacology and Toxicology Department NIPRD and Dr. Tijani A.Y,

for his contributions to my life both academically and morally during my six months of

attachment in the department. Not forgetting Mrs Augustina Nwachukwu for her unending

support.

I wish to thank my fellow IT students for their cooperation and support. I wish to also appreciate

my school supervisor Dr Aina.

v

Table of Contents

Certification i

Dedication ii

Declaration iii

Acknowledgement iv

Table of Contents v

List of Figures vii

List of Tables viii

CHAPTER ONE 1

Introduction 1

1.0 INTRODUCTION 1

1.1 AIMS OF SIWES 1

1.2 AGENCIES OF SIWES 2

1.3 SAFETY PRECAUTIONS 2

CHAPTER TWO 4

Organization Background 4

2.1 History 4

2.2 VISION OF NIPRD 4

2.3 MISSION OF NIPRD 4

2.4 ORGANIZATIONAL STRUCTURES OF NIPRD 5

CHAPTER THREE 8

Summary of Task Assigned 8

3.0 COMPARATIVE FEED ANALYSIS STUDY 8

Abstract 8

Summary 8

Materials 8

Laboratory Apparatus 9

Methodology 13

Procedures 15

Results 16

Discussion 26

Conclusion 28

vi

3.1 PILOT STUDIES FOR THE USE OF METABOLIC AND ORDINARY CAGES

(weighing of the feed, water and weight of Albino rats) 29

PURPOSE 29

MATERIALS 30

PROCEDURE 30

RESULTS 30

PRECAUTIONS 32

CONCLUSION 33

3.2 PSYCHOPHARMACOLOGY TEST (TESTING ANXIETY IN MICE USING EPM)

34

AIM 34

METHOD 34

PROCEDURE 35

OBSERVATION 35

3.3 ACUTE OVERDOSE EFFECT OF MORINGA IN MICE 36

BACKGROUND 36

EXPERIMENTAL 36

RESULTS 37

DISCUSSION 37

CONCLUSION 37

CHAPTER FOUR 38

4.0 SUMMARY OF ATTACHMENT ACTIVITIES 38

4.1 PROBLEMS ENCOUNTERED DURING ATTACHMENT 38

4.2 Conclusion 38

References 40

Appendix 41

vii

List of Figures

Figure 3. 1: Metabolic Cage [2] 9

Figure 3. 2: Y- Maze Apparatus [3] 10

Figure 3.3: Elevated-Plus Maze [4] 10

Figure 3.4: Light/Dark Box Apparatus [5] 11

Figure 3.5: The Hole-Board Apparatus [6] 11

Figure 3.6: Open Field Box Apparatus [7] 12

Figure 3. 7: Tail Suspension Box Apparatus [8] 12

Figure 3. 8: Facial Mask 15

Figure 3. 9: Growth Performance in Mice 25

Figure 3. 10: Growth Performance in Rats 25

Figure 3. 11: Rat in a Metabolic Cage 29

Figure 3. 12: Studying the Albino Mice using the Metabolic Cage 32

Figure 3.13: Mice in EPM 34

Figure 3. 14: Studying the Albino Mice Using the EPM 35

viii

List of Tables

Table 3. 1: Grouping of the Rodents 13

Table 3. 2: Feeds and Measurement 14

Table 3. 3: Body Weight of Rodents 16

1

CHAPTER ONE

Introduction

1.0 INTRODUCTION

Student industrial work experience scheme (SIWES) was designed by the federal government

and was being implemented by the industrial training fund (ITF) to bridge the gap between

theoretical and practical aspect of ones course of study due to the fact that the knowledge

gotten from various institution are more theoretical.

Nigerian graduates of tertiary institutions established SIWES in 1973 to solve the problem of

lack of adequate practical skills preparatory for employment in industries.

The scheme exposes students to industry-based skills necessary for a smooth transition from

the classroom to the world of work. It affords students of tertiary institutions the opportunity

being familiarized and exposed to the needed experience in handling machinery and equipment,

which are usually not available in the educational institutions.

1.1 AIMS OF SIWES

The aims of SIWES are:

Provide an avenue for students in institutions of higher learning to acquire industrial

skills and experience in their approved course of study.

Prepare students for the industrial works situation which they are likely to meet after

graduation.

Enlist and strengthen employers involvement in the entire educational process and

prepares student for employment in industry and commerce (Information and

guidelines for SIWES, 2002).

2

1.2 AGENCIES OF SIWES

Participation in SIWES has become a necessary pre-condition for the award of Diploma and

Degree certificates in specific disciplines in most institutions of higher learning in the country,

in accordance with the education policy of government.

OPERATORS: The ITF, the coordinating agencies (NUC, NCCE, NBTE), employers of

labour and the institutions.

THE SUPERVISING AGENCIES (NUC, NBTE, and NCCE): These agencies are to:

I. Ensure the establishment and accreditation of SIWES units in institutions under their

jurisdiction.

II. Direct for the appointment of full-time SIWES coordinators.

III. Ensure adequate funding of the SIWES units in all the institutions.

FUNDING: The Federal Government of Nigeria.

BENEFICIARIES: Undergraduate students of the following: Agriculture, Engineering,

Technology, Environmental, Science, Education, Medical Science and Pure and Applied

Sciences.

DURATION: Four months for Polytechnics and Colleges of Education and six months for the

Universities.

(ITF: Industrial Training Fund) (NUC: National Universities Commission) (NCCE: National

Commission for Cooperative Education) (NBTE: National Board for Technical Education)

1.3 SAFETY PRECAUTIONS

There are rules and regulation governing the laboratory in order to avoid any problem. They

include:

You must always wear your laboratory coat before coming into the laboratory and taken

off before visiting rest rooms, recreation room, and canteens etc..

Used hypodermic needles must be placed in commercially available containers with

puncture-proof, imperforate walls. When full these containers must be incinerated.

Always wash your hands before going out of the laboratory.

3

Consider any instrument in the laboratory hazardous and be careful handling them.

No making of noise, smoking, and eating in the laboratory..

All working surfaces should be disinfected and cleaned after use.

All specimen containers must be sterilized by autoclaving at 1210c for 15 minutes

Eye Glasses and Hand Gloves should be worn during blood collection.

You must not go out of the laboratory with Hand gloves on your hands.

Do not sit on laboratory benches.

You must wear closed shoes and not walking barefoot.

No unauthorized experiments are to be performed. If you are curious about trying a

procedure not covered in the experimental procedure, consult your laboratory instructor

4

CHAPTER TWO

Organization Background

2.1 History

BRIEF HISTORY OF NATIONAL INSTITUTE FOR PHARMACEUTICAL

RESEARCH AND DEVELOPMENT (NIPRD).

The institute was established under the science and technology act of 1980 with the primary

objective of developing drugs, biological products and pharmaceutical raw materials from

resources found in Nigeria.

NIPRD was established in 1988 and became operational in 1989 with the joint efforts of the

pharmaceutical society of Nigeria (PSN) and the federal government of Nigeria solely for

pharmaceutical research and development.

NIPRD has also developed medicinal products for the treatment and management of diseases

such as HIV/AIDS, malaria, TB, diabetes and SCD, which are prevalent in our society. NIPRD

is building capacity for the synthesis of first line retroviral and anti-malaria agents.

2.2 VISION OF NIPRD

To build a centre of excellence, for research and development of phytomedicines,

pharmaceutical and biological products for the services of mankind.

2.3 MISSION OF NIPRD

I. To apply appropriate modern sciences and technological resources to stimulate

local production of drugs through effective collaboration with the industry and

experts within and outside Nigeria.

II. Developing herbal products and phytomedicines to pilot stage for

commercialization.

III. Develop quality standard for drugs, herbal products phytomedicines, and

diagnostic for the purpose of control and regulation.

5

IV. Provide quality assurance services on drugs used in healthcare delivery as well

as herbal products and phytomedicines registered by the regulatory authorities

2.4 ORGANIZATIONAL STRUCTURES OF NIPRD

I. NIPRD consist of five (5) departments which are:

II. Medicinal plant research and traditional medicine

III. Medicinal chemistry and quality control

IV. Pharmacology and toxicology

V. Pharmaceutical technology and raw material development

VI. Microbiology & biotechnology

6

NIPRDS ORGANOGRAM

Legal unit

Protocol public relation unit

NIPRD research

clinic

Consultancy unit

DIRECTOR

GENERAL/CEO

GOVERNING BOARD

HONOURABLE

MINISTER

ICT, library, information & documentation unit

Director

medicinal plant

research and

traditional

medicine

department

Director

pharmaceutical

technology & raw

material

development

department (PT&RMD)

Audit unit

Procurement unit

Servicom unit

Director

finance&

accounts

Director

human

virology

&biotechnolog

y department

Director

medicinal

chemistry &

quality

control department

Director, pharmarmacco

lgy&toxicology

department M & e knowledge mgt

DEPUTY

DIRECTOR

Accounts

DEPUTY

DIRECTOR

Budget

DEPUTY

DIRECTOR

Finance DEPUTY

DIRECTO

R APDC

DEPUTY

DIRECTOR

Photoche

mistry

DEPUTY

DIRECTOR

pharmacognsy

traditional

medicine

DEPUTY

DIRECTOR

Biotechnolog

y

DEPUTY

DIRECTOR

human virology

DEPUTY

DIRECTOR

pharmaceutica

l technology

division

DEPUTY

DIRECTOR

raw materials

development

DEPUTY

DIRECTOR

Toxicology /

safety studies

DEPUTY

DIRECTOR

Pharmacolog

y

DEPUTY

DIRECTOR

GS

DEPUTY

DIRECTOR

Staff

welfare &

training

DEPUTY

DIRECTOR

medicinal

chemistry and

quality systems

development.

DEPUTY DIRECTOR

Biopharmaceuticals &

dug metabolism/

pharmacokinetics

(DMPK)

7

DEPARTMENT OF PHARMACOLOGY AND TOXICOLOGY

The department is divided into

Animal facility Centre

In vivo and in vitro laboratory

Whole animal studies laboratory

Plant collection section.

FUNCTIONS

Plant identification, collection and preservation

Acute toxicology studies

Chronic toxicology studies

Laboratory Animal breeding

Drug test and authentication

8

CHAPTER THREE

Summary of Task Assigned

3.0 COMPARATIVE FEED ANALYSIS STUDY

A comparative study of 3 different feed formula. (Effects on weight gain, growth performance and

mortality reduction of albino rats and mice)

Abstract

This study was carried out in the animal faculty centre. The comparison was between three

different feed formulas that was made; FF, CS and SF (control) and the use of rodents. Mice and

Rat was used as rodents during this experiments. There was total number of 48 rodents, 24 rats

and 24 mice, male and female inclusive.

Summary

The different feed formula were made up of different nutritional components individually and were

used to feed these rodents on daily basis. These rodents were selected randomly regardless of sex

and size (weight). Time was taken to select these rodents to ensure that the heathier ones were

selected over the weaker ones.

Materials

10 ordinary Cages

Weighing Balance

Hand Gloves

Face Mask

3 Different Feeds

Water

Book and Pen

48 albino mice and rats

9

Laboratory Apparatus

Laboratory apparatus is the individual instruments or pieces of equipment, or the entire set of

equipment, used in homes and labs to conduct projects and experiments. They are the most

common utensils and appliances that you need while performing hands on activities in a laboratory.

[1] Below are list of apparatus found in the pharmacology laboratory at NIPRD

Metabolic Cage: are for rodents weighing less than 300 grams. These Metabolic Cages include

four fluid collection tubes with funnels (two water spillage tubes and two urine collection tubes),

two faeces collection tubes, two urine rings and a feeder chamber and drawer sized for the animal

specified. The Cage for Mice also has mouse-size support grid and urine collection tubes. The

Diuresis Cage for Mice is identical in size to the Metabolic Cages but has a smaller separating

cone and collection funnel to minimize evaporation. It is used to collect by-products from a single

mouse. [2]

Figure 3. 1: Metabolic Cage [2]

10

Y- maze: is a simple maze used in animal cognition experiments. It is shaped like the letter Y,

providing the subject, typically a rodent, with a straightforward choice. Y-mazes are used to study

how the rodents function with memory and spatial learning through applying various stimuli.

Starting in the early 20th century, rodents were used in experiments such as the Y-Maze. These

concepts of Y-mazes are used to assess rodent behaviour. The different tasks, such as left-right

discrimination and forced alternation, are mainly used with rodents to test reference and working

memory. [3]

Figure 3. 2: Y- Maze Apparatus [3]

Elevated Plus Maze: The standard elevated plus-maze is commonly used to assess anxiety-like

behaviour in laboratory animals (rats/mice). The maze is usually a cross-shaped maze with two

open arms and two closed arms, which is elevated above the floor. [4]

Figure 3.3: Elevated-Plus Maze [4]

11

Light/Dark Box: The Light/Dark Box is a characteristic tool used in the assessment of anxiety.

The basic measure is the animals preference for dark, enclosed places over bright, exposed places.

Time spent in the lit (light) half of the arena, and the related exploratory behaviours, are reliable

parameters for assessing anxiolytic effects that may be useful in identifying and/or screening of

anxiolytic and anxiogenic agents. [5]

Figure 3.4: Light/Dark Box Apparatus [5]

Hole Board: The hole-board apparatus consists of an enclosed space, the floor of which has sixteen

holes in a grid-pattern. [6] The rodent, when placed in the apparatus, is free to dip its head through

the holes in the floor; the frequency and duration of this behaviour, known as 'head-dipping' is

thought to measure levels of neophilia. [6]

Figure 3.5: The Hole-Board Apparatus [6]

12

Open Field: The Open Field apparatus is broadly used to assess exploratory behaviour and is

validated for use in the measurement of anxiety related behaviours. [7]

Figure 3.6: Open Field Box Apparatus [7]

Tail Suspension Box: The tail-suspension test is a mouse behavioural test useful in the screening

of potential antidepressant drugs, and assessing of other manipulations that are expected to affect

depression related behaviours. Mice are suspended by their tails with tap [8]

Figure 3. 7: Tail Suspension Box Apparatus [8]

13

Methodology

The study was done using the 24 rats and mice respectively. The mice and rats were coded with

puric acid- the acid was used to label parts of their body for easy identification. For example, an

animal labelled on the head was tagged HD while the animal labelled on the tail was tagged TL.

Animals with label on both head and tail were tagged HDTL. The coding was done for all 48

rodents. Furthermore, their initial weight was being recorded with the aid of a weighing balance.

Thereafter, the grouping process was initiated; a total number of 10 ordinary cages were used for

the grouping of the 48 rodents.

Table 3. 1: Grouping of the Rodents

S/N No of

Cages

Rodents Sex Rodent

per cage

Feeds

1 1 Rats Male 5 Rats FF

2 1 Rats Female 5 Rats FF

3 1 Rats Male 5 Rats CS

4 1 Rats Female 5 Rats CS

5 1 Rats Control 4 Rats SF

6 1 Mice Male 5 Mice FF

7 1 Mice Female 5 Mice FF

8 1 Mice Male 5 Mice CS

9 1 Mice Female 5 Mice CS

10 1 Mice Control 4 Mice SF

Total Number of Cages = 10

Total Number of Male Rodents attached to FF = 10

Total Number of Male Rodents attached to CS = 10

Total Number of Female Rodents attached to FF = 10

Total Number of Female Rodents attached to CS = 10

14

Total of Number of Control Rodents attached to SF = 8

Total Number of Rodents = 48

Table 1.1 clearly shows how the grouping of the rodents was done.

The feeding aspect is one of the most significant aspect in this experiment. The feeding was done

three times a week; Monday, Wednesday and Friday respectively. The table below shows the

details of feed given to the rodents.

Table 3. 2: Feeds and Measurement

S/N

RATS

Feeds Measurement Sex

Mice

Feeds Measurement Sex

1 FF 400g Male FF 200g Male

2 FF 400g Female FF 200g Female

3 CS 400g Male CS 200g Male

4 CS 400g Female CS 200g Female

5 SF 320g Control SF 160g Control

The measurement column in the table above depicts the standard measure of feed given to these

rodents with their weight in consideration.

Furthermore, the 3 different feed formula that was introduced were in dry moulded form. The CS

feed and FF feed were moulded dry and looked like a chunk of mass with little or no H2O in it.

They were oven-dried while the SF feed came in a small cylindrical form, long and dry as well.

They could be easily described as a piece of chalk (pelleted) with a very brown colour. Also, the

feed FF had the same colouration, dark brown but feed CS had different colour entirely, cream

colour. These feeds were bagged differently and stored in the animal centre with each bag carefully

labelled.

Basically, these rodents were fed each day with these feeds. Their weight was recorded every two

days to keep record of the growth rate. However, leftover from the feeds were also weighed in

order to determine the consumption rate. Each rodents was provided with feed and water.

15

Procedures

The feed was 400 grams measure, 200 grams for the mice, 320 for rats (control) and 160 grams

for mice (control)

The animals were weighed on a 2 days interval alongside their feed, the readings were recorded

so as to get the weekly average. The fresh feeds were also weighed and replaced.

Figure 3. 8: Facial Mask

16

Results

Table 3. 3: Body Weight of Rodents

MICE

DATE

FF-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 HD 19 20 21.8 22.9 21.74

2 TL 15 13 16.9 18.1 16.6

3 LS 21 22 23.4 23.8 23.04

4 LL 17 18 21.7 22.3 20.4

5 RH 17 18 20.3 20.6 19.58

FEMALES 18.472

6 HDRS 21 19 20.9 22.1 20.6

7 NKRH 20 19 21.6 22.0 20.12

8 BKLS 17 17 18.3 18.0 17.26

9 NKLL 23 23 25.8 27.0 23.76

10 NM 19 19 21.2 22.1 20.06

20.36

MICE

B2

DATE

CS-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 BK 22 20 20.6 21.0 20.8

2 RS 21 20 19.4 19.0 19.68

3 RL 14 13 14.3 15.0 14.25

4 NK 17 16 16.4 17.0 16.68

5 LH 16 15 16.6 17.0 16.32

FEMALES 17.522

6 HDTL 23 22 21.6 22.0 22.12

7 LLRH 17 16 17.1 17.0 17.02

8 LLLH 19 17 17.1 17.0 17.42

9 BKHD 21 19 19.2 19.0 19.44

10 RLRS 18 17 16.9 17.0 17.38

18.67

17

MICE

MICE

DATE

FF-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID BODY

WT

2

BODY

WT

3

BODY

WT

4

BODY

WT

BODY WT

1 HD 19 20 21.8 22.9 21.74

2 TL 15 13 16.9 18.1 16.6

3 LS 21 22 23.4 23.8 23.04

4 LL 17 18 21.7 22.3 20.4

5 RH 17 18 20.3 20.6 19.58

FEMALES 18.472

6 HDRS 21 19 20.9 22.1 20.6

7 NKRH 20 19 21.6 22.0 20.12

8 BKLS 17 17 18.3 18.0 17.26

9 NKLL 23 23 25.8 27.0 23.76

10 NM 19 19 21.2 22.1 20.06

20.36

MICE

MICE

B2

DATE

CS-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID 1

BODY

WT

2

BODY

WT

3

BODY

WT

4

BODY

WT

BODY WT

1 BK 22 20 20.6 21.0 20.8

2 RS 21 20 19.4 19.0 19.68

3 RL 14 13 14.3 15.0 14.25

4 NK 17 16 16.4 17.0 16.68

5 LH 16 15 16.6 17.0 16.32

FEMALES 17.522

6 HDTL 23 22 21.6 22.0 22.12

7 LLRH 17 16 17.1 17.0 17.02

8 LLLH 19 17 17.1 17.0 17.42

9 BKHD 21 19 19.2 19.0 19.44

10 RLRS 18 17 16.9 17.0 17.38

18.67

18

RATS

RATS

DATE

FF-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID 1

BODY

WT

2

BODY

WT

3

BODY

WT

4

BODY

WT

BODY WT

1 HD 71 68 76.6 82.3 78.18

2 BK 109 116 130.3 137.2 128.5

3 TL 87 88 99.2 105.5 98.94

4 RL 88 86 100.2 105.3 98.7

5 LH 53 55 66.6 75.1 66.54

FEMALES

6 HDTL 84 89 101.2 105.7 98.38

7 LLRH 95 97 108.5 112.4 105.78

8 NKLL 58 58 64.7 68.8 64.5

9 HDRS 51 52 58.8 63.7 58.7

10 RSLS 72 75 82.5 87.5 82.8

20.36

RATS

RATS

B1

DATE

CS-MALE

0 1

22/01/15

1 2

23/01/15

2 3 4

26/01/15

3 4 6

28/01/15

AVG/WK

S/NO ID 1

BODY

WT

2

BODY

WT

3

BODY

WT

4

BODY

WT

BODY WT

1 RS 93 87 85.2 86.1 87.86

2 LS 103 100 100.3 98.4 100.54

3 LL 88 78 81.9 84.1 66.41

4 NK 83 75 73.4 73.7 76.22

5 RH 78 74 76.2 78.7 77.4

FEMALES

6 LLLH 119 113 119.4 124.5 120.78

7 RHLH 79 75 78.9 80.4 79.06

8 NKRH 63 60 60.4 62.4 62.36

9 BKHD 60 55 54.7 53.7 55.88

10 BKLS 50 48 48.7 48.7 48.88

19

MICE

MICE

DATE

FF-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID 1

BODY WT

2

BODY WT

3

BODY WT

4

BODY WT

BODY WT

1 HD 25.0 26.0 27.0 27.0 26.66

2 TL 20.0 22.0 23.0 24.0 23.00

3 LS 25.0 25.0 25.0 27.0 25.66

4 LL 23.0 25.0 25.0 22.0 24.00

5 RH 22.0 22.0 23.0 23.0 22.66

FEMALES

6 HDRS 20.0 24.0 24.0 25.0 24.33

7 NKRH 18.0 24.0 23.0 24.0 23.66

8 BKLS 16.0 21.0 21.0 22.0 21.33

9 NKLL 22.0 27.0 28.0 28.0 27.66

10 NM 19.0 24.0 25.0 25.0 24.66

MICE

MICE

DATE

CS-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 BK 21.90 21.0 21.0 22.0 21.33

2 RS 19.0 20.0 20.0 21.0 20.33

3 RL 15.0 15.0 15.0 16.0 15.33

4 NK 17.0 17.0 18.0 19.0 18.0

5 LH 17.0 19.0 19.0 20.0 19.33

FEMALES

6 HDTL 22.0 22.0 22.0 23.0 22.33

7 LLRH 18.0 18.0 18.0 18.0 18.00

8 LLLH 17.0 18.0 18.0 17.0 17.66

9 BKHP 19.0 19.0 20.0 19.0 19.33

10 BLRS 18.0 18.0 19.0 20.0 19.00

20

RATS

RATS

DATE

FF-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 HD 93.0 102.0 115.0 120.0 112.33

2 BK 150.0 158.0 169.0 174.0 167.00

3 TL 114.0 120.0 127.0 130.0 125.66

4 RL 114.0 125.0 132.0 138.0 131.66

5 LH 83.0 89.0 98.0 100.0 95.66

FEMALES

6 HDTL 112.0 114.0 118.0 122.0 118.00

7 LLRH 116.0 122.0 123.0 125.0 123.33

8 NKLL 73.0 80.0 85.0 94.0 86.33

9 HDRS 68.0 72.0 75.0 76.0 74.33

10 RSLS 97.0 103.0 110.0 114.0 109.00

RATS

RATS

DATE

CS-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 RS 88.0 87.0 86.0 83.0 85.33

2 LS 101.0 98.0 97.0 94.0 96.33

3 LL 85.0 85.0 87.0 87.0 86.33

4 NK 76.0 77.0 77.0 80.0 78.00

5 RH 80.0 84.0 86.0 90.0 86.66

FEMALES

6 LLLH 128.0 127.0 131.0 128.0 128.66

7 RHLH 82.00 83.0 84.0 80.0 82.33

8 NKRH 66.0 66.0 65.0 65.0 65.33

9 BKHD 56.0 53.0 56.0 53.0 54.00

10 BKLS 49.0 48.0 48.0 46.0 47.33

21

MICE

MICE

DATE

FF-MALE

8 15

06/01/15

9 18

09/02/15

10 20

11/02/15

11

13/02/15

AVG/WK

S/NO ID 1

BODY WT

2

BODY WT

3

BODY WT

4

BODY WT

BODY WT

1 HD 27.0 28.0 28.0 28.0 28.0

2 TL 24.0 26.0 26.0 27.0 26.3

3 LS 27.0 28.0 27.0 28.0 27.6

4 LL 22.0 27.0 26.0 28.0 27.0

5 RH 23.0 24.0 23.0 24.0 23.6

FEMALES

6 HDRS 25.0 25.0 26.0 27.0 26.0

7 NKRH 24.0 25.0 23.0 24.0 24.0

8 BKLS 22.0 23.0 22.0 22.0 22.3

9 NKLL 28.0 27.0 27.0 27.0 27.0

10 NM 25.0 26.0 25.0 25.0 25.3

MICE

MICE

DATE

CS-MALE

8 15

06/01/15

9 18

09/02/15

10 20

11/02/15

11

13/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 BK 22.0 22.0 22.0 22.0 22.0

2 RS 21.0 21.0 20.0 19.0 20.0

3 RL 16.0 15.0 16.0 15.0 15.3

4 NK 19.0 18.0 18.0 18.0 18.0

5 LH 20.0 21.0 21.0 20.0 20.6

FEMALES

6 HDTL 23.0 22.0 22.0 22.0 22.0

7 LLRH 18.0 18.0 18.0 18.0 18.0

8 LLLH 17.0 17.0 17.0 16.0 16.6

9 BKHP 19.0 19.0 20.0 20.0 19.6

10 BLRS 20.0 19.0 19.0 19.0 19.0

22

RATS

RATS

DATE

FF-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID 1

BODY WT

2

BODY WT

3

BODY WT

4

BODY WT

BODY WT

1 HD 120.0 123.0 128.0 132.0 127.6

2 BK 174.0 170.0 169.0 174.0 171.0

3 TL 130.0 134.0 142.0 145.0 140.6

4 RL 138.0 144.0 150.0 153.0 149.0

5 LH 100.0 105.0 111.0 114.0 110.0

FEMALES

6 HDTL 122.0 126.0 125.0 127.0 126.0

7 LLRH 125.0 131.0 128.0 131.0 130.0

8 NKLL 94.0 94.0 95.0 96.0 95.0

9 HDRS 76.0 80.0 79.0 80.0 79.6

10 RSLS 114.0 123.0 122.0 128.0 124.3

RATS

RATS

DATE

CS-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID 1

BODY WT

2

BODY WT

3

BODY WT

4

BODY WT

BODY WT

1 RS 88.0 87.0 86.0 83.0 85.33

2 LS 101.0 98.0 97.0 94.0 96.33

3 LL 85.0 85.0 87.0 87.0 86.33

4 NK 76.0 77.0 77.0 80.0 78.00

5 RH 80.0 84.0 86.0 90.0 86.66

FEMALES

6 LLLH 128.0 127.0 131.0 128.0 128.66

7 RHLH 82.00 83.0 84.0 80.0 82.33

8 NKRH 66.0 66.0 65.0 65.0 65.33

9 BKHD 56.0 53.0 56.0 53.0 54.00

10 BKLS 49.0 48.0 48.0 46.0 47.33

23

MICE

MICE

DATE

FF-MALE

8 15

13/02/15

9 18

16/02/15

10 20

18/02/15

11

20/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 HD 31.0 30.0 32.0 31..0

2 TL 28.0 28.0 28.0 28.0

3 LS 29.0 29.0 28.0 28.6

4 LL 29.0 29.0 30.0 29.3

5 RH 25.0 25.0 25.0 25.0

FEMALES

6 HDRS 26.0 26.0 26.0 26.0

7 NKRH 25.0 25.0 25.0 25.0

8 BKLS 23.0 23.0 23.0 23.0

9 NKLL 28.0 28.0 28.0 28.0

10 NM 25.0 25.0 26.0 25.3

MICE

MICE

DATE

CS-MALE

8 15

13/02/15

9 18

16/02/15

10 20

18/02/15

11

20/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 BK 22.0 22.0 21.0 21.6

2 RS 20.0 20.0 18.0 19.3

3 RL 16.0 16.0 15.0 15.6

4 NK 19.0 18.0 16.0 17.6

5 LH 21.0 21.0 18.0 20.0

FEMALES

6 HDTL 22.0 22.0 22.0 22.0

7 LLRH 18.0 18.0 18.0 18.0

8 LLLH 17.0 17.0 17.0 17.0

9 BKHP 20.0 20.0 21.0 20.3

10 BLRS 20.0 21.0 21.0 20.6

24

RATS

RATS

DATE

FF-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 HD 139.0 142.0 145.0 142.0

2 BK 180.0 186.0 188.0 184.6

3 TL 154.0 160.0 167.0 160.3

4 RL 159.0 162.0 168.0 163.0

5 LH 119.0 121.0 128.0 122.6

FEMALES

6 HDTL 131.0 132.0 136.0 133.0

7 LLRH 133.0 131.0 132.0 132.0

8 NKLL 99.0 104.0 109.0 104.0

9 HDRS 84.0 88.0 90.0 87.3

10 RSLS 135.0 138.0 142.0 138.3

RATS

RATS

DATE

CS-MALE

5 8

30/01/15

6 11

02/02/15

7 13

04/02/15

8

06/02/15

AVG/WK

S/NO ID BODY WT BODY WT BODY WT BODY WT BODY WT

1 RS DEAD DEAD DEAD

2 LS DEAD DEAD DEAD

3 LL 90.0 86.0 87.0 87.6

4 NK 86.0 83.0 82.0 83.6

5 RH 96.0 93.0 93.0 94.0

FEMALES

6 LLLH 134.0 132.0 134.0 133.3

7 RHLH 83.0 81.0 81.0 81.6

8 NKRH 71.0 69.0 70.0 70.0

9 BKHD 60.0 59.0 58.0 59.0

10 BKLS DEAD DEAD DEAD DEAD

25

The table above accommodates the recordings of the rodents as to how they react to the feeds.

Records were taken down at an interval of two days on a weekly basis. The table makes it possible

for one to observe the changes occurred in the rodents body weight.

Figure 3. 9: Growth Performance in Mice

Figure 3. 10: Growth Performance in Rats

0

5

10

15

20

25

30

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Growth Performance in Mice

FF CS

0

20

40

60

80

100

120

140

160

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Growth Perfomance in Rats

FF CS

26

Discussion

Figure 3.8 graphically shows the growth progression caused by the two feed, FF and CS

respectively. Notice that both categories had the same kick off point on the first recording but the

changes experienced during the second recording is quite obvious. Mice placed under the FF feed

experienced an immediate growth boost and their growth performance is obviously better than the

mice placed under the CS feed.

However, both categories experienced a sudden declination in their growth rate, FF seem to have

experienced a great declination but inclined rapidly as well. CS almost had a steady growth chart

with little more declination in the growth rate.

Having studied and analysed the growth chart, one could conclude by saying the FF feed had a

better impact on the growth of the mice than that of the CS feed.

The figure above clearly depicts the growth performance of rats under two different feeds, FF and

CS. Observing from the initial point, the CS feeds had the initial advantage as it started off on a

higher note than that of the FF feed.

However, the Growth performance for CS is certainly to poor as it depreciated instead of

appreciating. At a point, CS experienced a growth change of 88.5g, which is higher than 81.6g,

the initial body weight recorded but it later depreciated to a value of 86.4g. Unlike the FF Feed,

the growth performance was really great. The initial body weight recorded was 76.8g but the value

was projected later on to 140.5g.

Conclusively, it is safe to say that FF feed is better off than the CS feed as the growth performance

for rats placed under the CS feed was very poor.

Rodents maintained on diets of CS showed no diet-associated difference in food intake or weight

gain. This finding is consistent with the mice and rats. In addition, food intake did not change when

the type of rodent was switched from rat to mice. This lack of effect could indicate that the feed

lack the appropriate portion of nutrients to boost growth rate. However, when rats had access to

27

The growth graph shows that the FF feed contains a good quantity of protein nutrition.

Requirements of young rodents where substitution of protein like building nutrients corresponds

to their intensive growth.

The disharmony of nutrition composition of the CS feed can also have an unfavorable effect in the

breeding of rats like pet. Harmonic nutrition including optimal feeding technique has a great

relation to the increase of the welfare of these rodents.

28

Conclusion

From the tables and the graph, we see the comparison between the two feeds FF and CS. It was

observed that both mice and rats fed with the FF feed increased in weight and size.

On the other hand, rodents fed with CS were weaker and experience a poor progress in growth.

Death was recorded for four male rats and two female rats respectively.

From this, we concluded the FF feed to have a higher nutritional value with high protein amount

than other feeds.

29

3.1 PILOT STUDIES FOR THE USE OF METABOLIC AND ORDINARY CAGES

(weighing of the feed, water and weight of Albino rats)

ANIMALS

3 Clean rats (that have not been used for any previous study) were taken from the animal facility.

PURPOSE

The aim is to explore certain issues before undertaking large-scale study

METHOD

Experiment was performed using 3 rats coded 5,15, and 13 with their initial weight taken using

weighing balance. Each rat was placed in the designated metabolic cage also labelled 5 and 15

while 13 for ordinary cage. Animals were allowed to acclimatize to their environment and feed

for 13 days[from November 5th to 17th]/ Each rat was provided with feed and water. For the rats in

metabolic cages, powdered feed was used as recommended by the manufacturers of the cage. Large

pelleted feed was for the rat in ordinary cage.

Figure 3. 11: Rat in a Metabolic Cage

The principle of a metabolic cage is to confine a rodent in an enclosure with grid floor. The upper

chamber is made of smooth, gnaw-proof and noiseless polycarbonate which is transparent to admit

total room light and is placed on top of a funnel. The urine flows through the grid floor down into

30

the middle of funnel and into a collection vial. On the sides the feaces fall-down separately into

another vial. To avoid contamination, feeding and watering compartments are cleverly

incorporated outside the cage: In order to access the chamber containing the food drawer, the

animal has to enter it through a trap door. The water bottle is calibrated, as well as the subjacent

spilling bottle, to calculate the actual water intake of the animal. In case of a spillage, water is

prevented from entering the collection funnel.

MATERIALS

Two metabolic cages [Ugo Basile, Italy] an ordinary cage, weighing balance, gloves, Petri dish, syringe,

logbook and pen, water, feeds[large and small], 3 albino rats.

PROCEDURE

The feed was 50grams and water 200ml for the three cages. Every day the feed and water intake

were recorded to calculate the average for the week. Also fresh feeds and water were weighed and

replaced. The animal weight was recorded once in a week.

RESULTS

Table 1: Weight [in gram]

Animal

identification

Initial

weight

Nov 5 and

7

1st week

November

18

2nd week

November

25

3rd week

December

1

4th week

December

8

5th week

December

15

5 131 160 165 165 177 178

15 130 171 179 184 186 185

13 156 171 172 176 182 176

31

Table 2: Feed Intake [in gram]

Animal

identification

1st week

November

18

2nd week

November

25

3rd week

December

1

4th week

December 8

5th week

December 15

5 10 13 10 11 ?

15 14 17 13 12 ?

13 27 25 32 35 ?

Table3: Water Intake [in millilitres]

Animal

identification

1st week

November

18

2nd week

November

25

3rd week

Decembe

1

4th week

December

1

5th week December 15

5 31 18 16 15 ?

15 36 21 20 21 ?

13 35 23 19 22 ?

32

Figure 3. 12: Studying the Albino Mice using the Metabolic Cage

PRECAUTIONS

1. I ensured that all measurement of drinking bottles was one without covers.

2. The spilling water bottle was poured into the drinking bottle before measurement.

3. The feed given to the ordinary cage was long pelleted to avoid dropping through the wire

mesh.

4. Adequate sanitation was carried out i.e. washing the metabolic cage properly once a week

and changing beddings of the ordinary cage.

33

CONCLUSION

More metabolic cages should be employed as further proof of results of the pilot study.

The small pelleted feed was good for rats in the metabolic cage because they gained as much

weight as the one in ordinary cages.

34

3.2 PSYCHOPHARMACOLOGY TEST (TESTING ANXIETY IN MICE USING

EPM)

AIM

To measure anxiolytic effects of drug in rodents

ANIMALS

Twenty male rats were gotten from the Animal facility centre in NIPRD.

HUSBANDRY

The animals were housed in stainless steel metal cages. They were shared into a group of fives,

i.e. four cages

METHOD

Azidothymine (HIV drug) was dissolved in water, and also a solution of normal saline was

prepared, to be administered orally. The solution was to be used as control. An Elevated Plus Maze

was cleaned. Azidothymine were dosed per kilogramme body weight. After administration,

animals are placed in the Elevated plus maze for observation at intervals of 5mins

Figure 3.13: Mice in EPM

35

The EPM is a box made of wood which is made up of both opened and closed arms i.e. total of

four arms which enable easy observation.

PROCEDURE

Azidothymine was administered orally. All the animals in cage1 were dosed with normal saline,

which was administered as control. Animals were left for 60mins to allow the drug work. After

5mins, each animal was placed in the centre of the EPM. For every movement to and fro both

closed and opened, a tally is drawn and time taken was recorded using Stopwatch. This is done for

the next 5mins and then the animal is removed and the field is cleaned and disinfected before

placing another animal in for observation. The same procedure was carried out for the rest of the

animals.

OBSERVATION

While animals were placed in the EPM, they displayed the following behaviours; rearing, freezing,

continuous sniffing. The animals induced with higher dose of the drug evaluate anxiolytic effects

than those given lower dosage. Also, the animals were depositing watery faecal materials. The

same procedure was carried out for rats induced with Normal saline.

Figure 3. 14: Studying the Albino Mice Using the EPM

36

3.3 ACUTE OVERDOSE EFFECT OF MORINGA IN MICE

BACKGROUND

Moringa is the formulated and encapsulated freeze-dried aqueous root extract of Moringa Oleifera.

As a step towards eventual introduction into clinical use for management of many diseases, data

on oral acute toxicity profile of the product is required to obtain approval from regulatory agency

to embark on commercial production of Moringa. This study was therefore designed to obtain data

on oral acute toxicity of formulated Moringa.

PREPARATION OF MORINGA SOLUTION

The shell of capsule containing 250 mg of the active principle was opened and poured into a

mortar. The shell content was triturated into homogenous solution with warm distilled water (10

ml) to obtain a final stock concentration of 25 mg/ml.

ROUTE OF ADMINISTRATION

Appropriate volume of MORINGA solution was administered via oral route.

EXPERIMENTAL

The study was carried out in accordance with OECD guidelines 423 (OECD, 2001) for acute oral

toxicity testing of formulated Moringa. Three female non-pregnant mice were given 300 mg/kg

body weight of Moringa orally and were observed for signs of adverse effect and mortality if any

at 1h interval for 4hrs. In the absence of mortality after 48hours of Moringa administration.

Another set of three female mice was given 300 mg/kg body weight of the sample and was

observed for signs of toxicity and mortality. In the absence of mortality after 48hrs of Moringa

administration, the dose was scaled up to 2000mg/kg body weight in another fresh set of three

non-pregnant mice and were observed for signs of toxicity and mortality at 1h interval for 4hrs.

The dose of 2000 mg /kg body weight of Moringa was repeated after 48hours in another fresh set

of three non-pregnant female mice in the absence of mortality. They were also observed for signs

of toxicity and mortality at 1h interval for 4hrs. Subsequently, all the mice in the study were

observed daily for 14 days for possible manifestation of delayed toxicity.

37

RESULTS

There were no deaths or any signs of toxicity observed after oral administration of single doses of

the decoction of the roots of Moringaat any dose level up to the highest dose tested (5, 000 mg/kg).

Moringa did not produce significant change in behaviour, breathing, coetaneous effects, sensory

nervous system responses and gastrointestinal effects in the mice used. The median lethal dose

(LD50) of the Moringa was estimated to be greater than 5000 mg/kg body weight orally.

DISCUSSION

The results obtained from the current study showed that acutely Moringa did not produce adverse

effects and mortality at the doses tested. The absence of signs of toxicity and mortality at all doses

used suggests that Moringa can be classified as practically non-toxic acutely in mice via oral route

of administration according to Globally Harmonized System (Roll et al., 1986). It may therefore

be concluded that Moringa is safe acutely in mice.

CONCLUSION

The Moringa is practically non-toxic acutely up to a dose of 5000 mg /kg body weight orally in

mice.

38

CHAPTER FOUR

4.0 SUMMARY OF ATTACHMENT ACTIVITIES

During the six months of my attachment with pharmacology and toxicology in NIPRD. A lot of

experience was gained. Most of which was beneficial in my field of study and moral life. Some

of the experiences gained include:

Basic animal handling.

Various routes of drug administration (most especially orally).

Ability to carry out major bench work on behavioural studies (Epm and Y-maze).

Blood collection from laboratory animal.

Ability to carry out major chemical screening.

Proper Time Management.

Communication Skills.

4.1 PROBLEMS ENCOUNTERED DURING ATTACHMENT

At first it was quite uneasy to go at the pace of some researchers in the department. It was expected

of trainees to have adequate knowledge of some studies. So I had to search the net regularly on

studies to be carried out, as they threw questions at IT students during most studies

Furthermore, Finance was one of the major issue being faced. In our daily operation,money is

needed to help carry out specific activities . There was no allowance provided for the interns; this

was quite a big challenge as one would need finance for transportation and feeding.

4.2 Conclusion

Having gone through this process, I must say that the importance of industrial training is very

paramount. Its not just to train students on new skills but it is to actually prepare them for the

industry; the real life scenario. It also serves as a career starter for most students guiding them in

making a decision on which career path to take.

39

So far, I have gained much knowledge and I see it as a good foundation for my career. I now have

a taste of what the industry looks like and I am fully prepared to embark on the journey. With these

knowledge gained, chances has been created for improvements on the basis of my personal

commitment.

40

References

[1] Joshua M. Pearce, Building Research Equipment with Free, Open-Source Hardware. Science 337 (2012).

[2] Novak, C.M., Burghardt, P.R. and Levine, J.A., 2012, The use of a running wheel to

measure activity in rodents: Relationship to energy balance, general activity, and reward,

Neuroscience and Biobehavioral Reviews,

[3] Lalonde, R; Strazielle C (2008). "Relations between open-field, elevated plus-maze, and

emergence tests as displayed by C57/BL6J and BALB/c mice.".

[4] Shoji, H., Hagihara, H., Takao, K., Hattori, S., Miyakawa, T. (2012) T-maze Forced

Alternation and Left-right Discrimination Tasks for Assessing Working and Reference

Memory in Mice.

[5] Lalonde, R; Strazielle C (2008). "Relations between open-field, elevated plus-maze, and

emergence tests as displayed by C57/BL6J and BALB/c mice".

[6] Engin, E. and Treit, D. (2008). "The effects of intra-cerebral drug infusions on animals'

unconditioned fear reactions: A systematic review.". Prog. Neuro-Psychopharmacol Biol.

Psychiatry

[7] Careau, VC; Bininda-Emonds ORP, Ordonez G, Garland T, Jr. (2012).

[8] Cryan JF, Slattery DA. Animal models of mood disorders: Recent developments. Curr

Opin Psychiatry 2007.

41

Appendix