CHUKA UNIVERSITY

JANUARY-MARCH FIELD ATTACHMENT REPORT CARRIED OUT AT REGIONAL

VETERINARY INVESTIGATION LABORATORY, KARATINA

DURATION: 8 WEEKS

BY
NAME: JOEL GACHURI NJOGU

REGISTRATION NUMBER: EB5/43601/19

A FIELD ATTACHMENT REPORT SUBMITTED TO THE DEPARTMENT OF PHYSICAL

SCIENCES FOR FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF BACHELOR OF
SCIENCE BIOMEDICAL SCIENCE AND TECHNOLOGY OF CHUKA UNIVERSITY.

1
DECLARATION

I declare that, this field attachment report is my original work, and a product of activities that I have
carried out at Regional Veterinary Investigation Laboratory from 10TH JANUARY –4TH MARCH 2022
and it has not been submitted by any other student to any department or university for academic award or
any other purpose.

Name: JOEL GACHURI NJOGU Registration number: EB5/43601/19

SIGN………………………………………DATE…………………………………

2
RECOMMENDATION
This report has been submitted with our approval as the University supervisors

1. Name
Institution
Signature…………………………………Date…………………………………

2. Name
Institution
Signature…………………………………Date ……………………

3
DEDICATION
Dedicate this Work to Almighty God, CHUKA UNIVERSITY for giving me the chance to undertake my
attachment for the award of my degree and attachment institution I am indeed grateful for the opportunity,
I also dedicate this work to my family for their financial and moral support.

4
ACKNOWLEDGEMENT
My sincere gratitude goes to the Management of Karatina RVIL for the opportunity to be an attaché for
hand on training in the institution.
Highly appreciated are all the technical staff of all laboratory sections in RVIL for sharing in the
knowledge and bench-work techniques wholeheartedly.
My gratitude also goes to my Supervisor Mr. Kariuki for the assessment during the attachment period.
To my fellow attachees, thank you for making part of the team. The sky is the limit.

5
ABSTRACT.
Field attachment is a field of practical training experience that prepares trainees for the task they are
expected to perform on completion of their training. This report contains information which I gathered
during my individual training conducted at RVIL Karatina. The attachment period was two months where
I started on 10THJANUARY and on 4THMARCH 2022 marked the end of my attachment period. The
institution is located in Nyeri County along Nanyuki-Nairobi Highway, Jamii road, Opposite Karatiana
Sub-county Hospital. The Laboratory is committed in providing quality Veterinary attention to farmers in
the mandate region. The institution also participates in various ongoing research especially drug and
antibiotic resistance. The institution is in collaboration with other RVIL across the county, they are 8 such
Laboratories in Kenya. The institution also ventures majorly on improving the health of livestock in the
mandate region and to prevent emergence of diseases through surveillance and vaccination services in the
mandate region.

I was able to learn and gain skills knowledge and experience by engaging ourselves in all laboratory’s
sections despite the challenges such as difficulty in adapting to the new environment, financial problems
among many others. Through this experience I was able to prepare media, primary culture, secondary
culture, AST techniques, Parasitology operations e.g. egg warm count and pathological procedures under
minimal supervision.

6
LIST OF ACRONYMS AND ABBREVIATIONS
CVL----Central Veterinary Laboratories
RVIL—Regional Veterinary Investigation Laboratory
KARLO--Kenya Agricultural Research and Livestock Organaization
ILRI–International Livestock Research Institute
KEMRI–Kenya Medical Research Institute
KEBS–Kenya Bureau of Standards
EU---European Union
WHO---World Health Organisation
CDC---Centers for Disease Control
VBM---Valuable Biological Material
GLWP---Good Laboratory Work Practice
SOP---Standard Operating Procedure
PPE--- Personal Protective Equipment
MRT---Milk Ring Test
RBPT---Rose Bengal Plate Test
CFT---Complement Fixation Test
ELISA--- Enzyme Linked Immuno-sorbent Assay
NCD--- New Castle Disease
HI--- Haemagglutination Inhibition.
PCR--- Polymerase Chain Reaction
EPG---Egg Per Gram

7
TABLE OF CONTENTS

DECLARATION………………………………………………………………….. 2
RECOMMENDATION………………………………………………………….... 3
DEDICATION……………………………………………………………………... 4
ACKNOWLEDGEMENT…………………………………………………………. 5
ABSRACT………………………………………………………………………...... 6
ABREVIATIONS………………………………………………………………….. 8
CHAPTER ONE………………………………………………………………......... 9
1.1 BACKGROUND INFORMATION OF THE INSTITUTION…………....... 10
1.2 GOALS AND OBJECTIVES…………………………………………………... 10
CHAPTER TWO……………………………………………………………………. 11
2.0 LITERATURE REVIEW OF RVIL KARATINA……………………………. 11
2.1 OVER VIEW ORGANIZATION OF RVIL KARATINA……………………. 11
2.3 ORGANOGRAM…………………………………………………………….. 13
2.4 DUTIES AND RESPONSIBILITIES………………………………………….. 14
2.6 BIOSAFETY AND BIOSECURITY…………………………………………… 15
2.10 GOOD LABORATORY WORK PRACTICE…………………………………. 16
2.11 BAD LABORATORY PRACTICES (DON’TS)……………………………….. 16
CHAPTER 3………………………………………………………………………..... 18
3.1 METHDOLOGY AND INSTRUMENTATION………………………………… 18
3.2 INSTRUMENTATION………………………………………………………….. 20
CHAPTER FOUR…………………………………………………………………... 23
4.0 GENERAL OVERVIEW………………………………………………………. 23
4.1 MEDIA AND STERILIZATION………………………………………………... 22
4.2 BACTERIOLOGY UNIT………………………………………………………... 25
4.3 PARASITOLOGY UNIT………………………………………………………... 28
4.4 PATHOLOGY UNIT (POST MORTEM)………………………………………. 30
4.5 SEROLOGY SECTION………………………………………………………….. 32
4.6 BIOCHEMISTRY SECTION…………………………………………………….. 36

8
4.7 FIELD ACTIVITIES…………………………………………………………………... 36
CHAPTER 5………………………………………………………………………..….. 38
5.1 SKILL, TECHNIQUES AND LESSONS LEARNT AND THEIR RELEVANCE…….. 38
5.2 CONTRIBUTIONS MADE TO THE INSTITUTION……………………………… 38
5.3 PROBLEMS ENCOUNTERED…………………………………………………….. 38
5.4 RECOMMENDATION…………………………………………………………… 39
5.5 CONCLUSION……………………………………………………………………... 39
5.6 REFERENCE………………………………………………………………………. 39

9
CHAPTER ONE
1.1 Background information of the institution

Karatina RVIL was established with NORAD support and started its operation on 31 st January 1997. The
sponsorship lasted for five years ending on 31st December 1979. Its establishment was due to give
farmersgreater support in the diagnosis, investigation and control of disease associated with livestock in
order to sustain a viable vibrant livestock in the region. RVIL Karatina is amongst the 8 labs in Kenya.
This lab was put up to mitigate challenges farmers are undergoing. The geographical location is
centralized to cover the following counties Nyeri, Muranga, Kirinyaga, Embu, Tharaka Nithi, Meru,
Laikipia, Marsabit and Samburu .
Animals dealt with

 Cattle.
 Avian species
 Fish.
 Bees.
 Porcine species
 Miscellaneous e.g. camel
Diseases handled include:
• Mastitis.
• Brucellosis.
• Anthrax.
• Trypanosomiasis.
• Helminthiasis.
• Rabies.
And many others.

1.2 Goals and objectives

• To fully get exposure to diagnosis, treatment, and prevention of diseases.
 To have a deeper insight to Microbiology work as far as diagnostics of diseases is concerned.
• To have clear view of the link between class work and real practical work in the diagnostic
Laboratory.
• To understand epidemiology of certain diseases Central region.
• To get full exposure to post mortem cases
• To be fully exposed to field cases study and meat inspection.

10
Mandate of the lab
To generate, preserve and effectively disseminate information for veterinary clinical intervention and
early warning system for animal health in order to access global market for livestock, livestock products
and by-products.
Vision
To be a benchmark reference Veterinary investigation laboratory in East and Central Africa.
Mission
To provide, facilitate quality and timely laboratory services in order to effectively contribute in
prevention, control and eradication programmes of animal disease, pest and vector for increased food
safety and security as well as create employment, promote local and international trade and industrial
growth in a suitable environment.

CHAPTER TWO
2.0 LITERATURE REVIEW OF RVIL KARATINA
2.1 Over view organization of RVIL Karatina
RVIL Karatiana has been in operation for more than 37 years since it was established. RVIL Karatina has
been working in collaboration with well-wishers e.g. European Union and other stake holders in ensuring
quality veterinary services are provided to farmers in the mandate region.
RVIL Operates in a referral system to the Central Veterinary Laboratory located at Kabete. The facility in
its capacity diagnosed and managed 4500 cases of Bovine species, 3500 cases of Avian species, 2000
cases of Porcine species and 1300 cases of Ovine cases in the last one year.
Currently the laboratory operates through the following machinery:

 Specimen collection
 Specimen safe handling
 Specimen Transportation
 Specimen safe reception at the Laboratory
 Specimen safe storage
 Specimen Processing
 Report and validation of Laboratory results
 Prescription by the Doctor in Duty

11
RVIL Karatina with its own excellent system of operations which are in compliance to the ministry of
Agriculture, Livestock and Fisheries and in Collaboration with Ministry of Health, the laboratory internal
audit is performed in every three months to ensure compliance and the report given
Observers of global Veterinary and Human health e.g. WHO, Ministry of Health, KARLO,ILRI,
Ministry of Agriculture, Livestock and Fisheries supervises the operation of the laboratory ensuring
adherence to ISO requirements and Zero hazard and pollution emitance to the community
2.2 Overview of the organization
The laboratory is supervised by the Central Veterinary Investigation Laboratory located at Kabete,
Kiambu County through a referral system.
The Officer-in-charge (Deputy Director of Veterinary Services) is the head of the station. He mandates
the Deputy Officer-in-charge, Chief Veterinary Officer(s) and the Support Staff.
The senior Laboratory analyst reports directly to The Chief Veterinary Officer, he also mandates the
Laboratory analyst, the technologists and the Support staff.

12
 2.3 ORGANOGRAM

OFFICER-IN CHARGE RVIL

KARATINA
DEPUTY DIRECTOR OF VETERINARY
SERVICES

DEPUTY OFFICER-IN CHARGE

CHIEF VETERINARY OFFICER

CHIEF VETERINARY
OFFICER (S)

SENIOR LABORATORY ANALYST

SUPPORT STAFF

CHIEF LIVESTOCK HEALTH

ASSISTANT

LIVESTOCK HEALTH
ASSISTANT (S)

LABORATORY
LABORATORY ANALYST
TECHNOLOGIST

13
2.4 DUTIES AND RESPONSIBILITIES
Officer-in-charge

 Officer in charge of the station.

 General supervises and coordinates all Laboratory’s activities and ensures delivery of
Laboratory’s services.
 Management of all resourses; human and material.
 Preparation and Imlementation of the station workplan.
 Liaison officer with relevant agencies and external stakeholders.
Deputy Officer-in-charge

 Deputy officer-in-charge of the station.

 Head of RVIL Technical operations.
 Quality assurance and standards officer charged with establishing, improving and maintain
standards.
 Routine disease diagnosis, disease surveys, farm visits exensions and outreach programmes.
Chief Veterinary Officer

 Validation of Laboratory results and prescription of choice of drug to the client.

 Routine Pathology work.
 In charge of training students on attachment.
 Routine disease diagnosis, disease surveys, farm visits extensions and outreach programmes
Senior Laboratory Analyst

 Receive Laboratory results and validates, reports to the Chief Veterinary Officers
 Routine Laboratory officer in all sections.
 Charged with establishing, improving and maintaining stanards in the Bacteriology and Media
room.

Laboratory Technologists and Analyst

 Carries out routine laboratory diagnostic procedures and submit the results to the laboratory
analyst for Validation.
 Preparation of media and ensures availability of the media when required.
 Ensures zero hazard leakage or pollution in the laboratory.

14
2.5 ACTIVITIES CARRIED OUT DURING THE ATTACHMENT PERIOD.

OBJECTIVES OF THE ATTACHMENT.

• To fully get exposure to veterinary services such as diagnosis, treatment, and prevention of
diseases.
• To have clear view of the link between class work and real practical work in the diagnostic
veterinary practices
• To fully get exposure to several laboratory procedures carried out at the laboratory.
• To understand epidemiology of certain diseases.
• To get full exposure to post mortem cases.

2.6 BIOSAFETY AND BIOSECURITY

Laboratory biosafety
Laboratory Biosafety describes the containment principles, technologies and practices that are
implemented to prevent the unintentional exposure to pathogens and toxins or their incidental release.
With the objective of containment being to confine biohazards (pathogens and toxins) and prevent
potential exposure of the laboratory worker to infectious agents.

2.7 Components of biosafety include;

Equipment used and procedures
• Biosafety cabinets
• PPE
• Autoclaves etc.

 Transport equipments
 Decontamination chemicals and reagents
 Storage of samples and other lab materials.
 Administrative – Registers, forms: roles, rules and regulations
 Entry/Exit procedures
 Waste disposal procedures- liquids, solids
 Detailed spill procedures

15
2.8 Laboratory Biosecurity
Laboratory Biosecurity describes the protection, control and accountability for Valuable Biological
materials (VBM) within laboratories, in order to prevent their unauthorized access, loss, theft, misuse,
diversion or intentional release.
Components of Biosecurity include;
Physical security-access control and monitoring
Personnel management
Information security
Transport of Biological materials
Accident, injury and incident response plans
Training and practice drills
Security updates and re-evaluation

2.9 BIOSECURITY IN RELATION TO BIOSAFETY

Biosecurity is part of Biosafety
Biosecurity and Biosafety are components of Good Laboratory Work Practice (GLWP)
Many laboratories with good Biosafety programs have already integrated basic elements of Biosecurity.
Laboratory Biosecurity and Biosafety share a common goal: protecting the personnel and the
environment, keeping Valuable Biological Material (VBM) safely, securely inside the areas where they
are used and stored.
2.10 GOOD LABORATORY WORK PRACTICE
These are guidelines on technical methods designed to avoid or minimize the majority of laboratory
accidents, injuries, and work related infections caused by human error, poor laboratory techniques and
misuse of equipment.
GLWP do’s

 Wear proper PPE

 Decontaminate/sterilize all biological waste before disposal
 Disinfect surfaces before and after work
 Label all specimens in the lab.
 Keep inventory for all Biological agents
 Follow authorized SOPs
 Observe proper hand washing
 Know the location of all safety and emergency equipment and procedures used in the lab.
 Know the Fire drill procedures and Locations of all exits.
 Know the location of the closest telephone.

16
 Familiarize yourself with all procedures before doing the lab exercise.
 Report all accidents, hazards or chemical spills to the instructor.
 Tie back all long hair and remove dangling jewellry during lab work sessions.
 Always make sure that Electrical Equipment is turned in the off position before plugging it into
the socket.
 Handle all Lab animals with care.
 Use extreme care when handling sharp objects.
 Dispose all Chemicals, broken glasses and other lab materials into the proper containers.
 When heating liquids in a test tube always point the test tube away from other people and
yourself.
 Keep all material away from open flames
 Take care when handling hot glassware and apparatus in the lab.
 Read labels and Equipment instructions carefully before use.
 Label all reagents/chemical properly (include date prepared, expiry date, name of person who
prepared & storage temperatures)
 Document all procedures
 Keep lab doors locked always
 Authorize all samples going out of lab.
 Avoid overcrowding in the lab
 2.11 BAD LABORATORY PRACTICES (DON’TS)
 Don’t shake hands in the lab
 Don’t eat/drink, smoking in the lab.
 Don’t pour Biological waste in the drain.
 Don’t take contaminated material (PPE) outside the Lab.
 Don’t work alone in the lab
 Avoid overcrowding in the lab.
 Do not mouth pipette.
 Never leave the laboratory without washing your hands.
 Never return unused reagents/chemicals to the original container.
 Never mix the reagents /chemicals before asking the instructor.
 Never use electrical Equipment around water.
 Never work in the Lab alone
 Never smell, taste or touch chemicals.
 Don’t wear contacts in the lab without proper eye protection.
 Never add water to an acid.
 Don’t chew gums or eat candy during lab exercise
 Never experiment on your own.
 Don’t engage in practical jokes and boisterous conduct in the lab.
 The use of personal audio or video equipment is prohibited in the lab.
 Don’t sit on the lab benches.
 Never leave the equipment after working.
 Don’t run in the lab.

17
CHAPTER 3

3.1 METHDOLOGY AND INSTRUMENTATION

METHODOLOGY:
Training and Observation.
During my attachment period, training was the major way of learning the skills in my area of study. The
training and observation of how the laboratory carries out its daily activities and how the staffs conduct
themselves, was the first thing to be taught by our supervisor

I received the following training during the entire attachment period.

• How to receive and reject samples from farmers.
• How to run various test on different samples submitted.
• How to operate, run and maintain various machines
• How to prepare microscopic structure slides and all the work that involves microscopy.
• How to enter results of various test into LIMS.
• How to prepare media, culturing of samples and interpretation of results.
Guidance and Demonstration.
Before engaging in any hands-on activities, the supervisor started by demonstrating the activity to us then
we could carry out the activity on our own. The supervisor guided us in activities that required high
skilled personnel and extra caution. Those activities that didn’t require most of practical work the
supervisor would guide us through on how to do it until the task is complete.

3.2 INSTRUMENTATION
Instrumentation refers to the tools or means used by a laboratory personnel to carry out their routine
activities in laboratory in order to achieve its goals and objectives. The instruments that I interacted with
are as follow; `

3.2.1 Biosafety/Biological Cabinet

Is an enclosed, ventilated equipment in a laboratory for safely working with materials contaminated with
pathogens that requires serious handling, the most recommended biosafety cabinet for use is Class 2b that
protects the environment, technician and the sample being handled.

18
Principle
The principle of operation of the cabinet involves a fan mounted in the top of the cabinet that draws a
curtain of sterile air over the workstation where the biological products are being handled. The air then
moves underneath the work station and back up to the top of the cabinet before passing through HEPA
filters. The exhaust that moves out of the facility consist of air being drawn into the front of the cabinet
underneath the work station.
The working mechanism of Class II BSCs differs according to its types. It has an open front with inward
airflow for personnel protection, downward HEPA filtered laminar airflow over the work surface for
product protection and HEPA filtered exhausted air for environmental protection. The room air and
recirculated air are HEPA filtered before flowing downwards over the work area. Class II BSCs can be
exhausted into the containment zone or directly to the outside atmosphere through a thimble or hard-
ducted connection depending on the types. The amount of air that recirculates or exhausts depends on the
types.
3.2.2 Autoclave
An autoclave is a machine that uses steam under pressure to kill harmful bacteria, viruses, fungi, and
spores on items that are placed inside a pressure vessel. The items are heated to an appropriate
sterilization temperature for a given amount of time. The moisture in the steam efficiently transfers heat
to the items to destroy the protein structure of the bacteria and spores.
3.2.3 Pressure cooker
A good alternative for the autoclave is the pressure cooker, which is based on the same principle: steam
accumulation, higher pressure and higher temperature. In contrast to the autoclave with downward
displacement of air, the pressure cooker operates with upward displacement of air by steam. Pressure
cookers are much cheaper than autoclaves.
Working Principle
The autoclave works on principle of moist heat sterilization. The high pressure inside the chamber
increases the boiling point of water for the sterilization of equipment. The higher pressure also ensures the
rapid penetration of heat into the deeper parts of equipment. The moisture present in the steam causes
coagulation of proteins of microbes causing irreversible loss of their activity and functions.
Working
Conditioning Phase (C): Air inhibits sterilization and must be removed from the chamber during the first
phase of the sterilization cycle known as conditioning. In dynamic air removal-type steam sterilizers, the
air can be removed from the chamber using a vacuum system. It can also be removed without a vacuum
system using a series of steam flushes and pressure pulses. Gravity-type sterilizers use steam to displace
the air in the chamber and force the air down the sterilizer drain.
Exposure Phase (S): After the air is removed, the sterilizer drain closes and steam is continuously
admitted into the chamber, rapidly increasing the pressure and temperature inside to a predetermined
level. The cycle enters the exposure phase and items are held at the sterilization temperature for a fixed
amount of time required to sterilize them.

19
Exhaust Phase (E): During the final phase of the cycle, exhaust, the sterilizer drain is opened and steam is
removed, depressurizing the vessel and allowing the items in the load to dry.
3.2.4 Hot Air Oven
A hot air oven, laboratory instrument that uses dry heat to sterilize laboratory equipment and other
materials. Equipment cannot be wet or material that will not melt, catch fire, or change form when
exposed to high temperatures are sterilized by using the dry heat sterilization method.
Working Principle
Electrical devices work on the principle of dry and hot air convection, conduction, and radiation. The hot
air convection process is of two types. a. Gravity convection process: Heated air expands and possesses
less density than cooled air which rises up and displaces the cooler air (the cooler air descends). It
produces inconsistent temperature within the chamber thus has a slow turnover. b. Mechanical
convection: Use of fitted blower or fan that actively forces heated air throughout all areas of the chamber.
This dry heat destroys bacterial endotoxins (or pyrogens) which are difficult to eliminate by other means.
This property makes it applicable for sterilizing glass bottles that are to be filled aseptically.
Dry heat kills by oxidation, protein denaturation, and toxic effects of elevated levels of electrolytes and it
is more efficient.
3.2.5 Centrifuge
Centrifuges are used in various laboratories to separate fluids, gases, or liquids based on density. In
research and clinical laboratories, centrifuges are often used for cell, organelle, virus, protein, and nucleic
acid purification.An example of centrifuge use in a clinical setting is for the separation of whole blood
components. Different assays necessitate serum or plasma, which may be obtained with centrifugation.
Serum is obtained by letting a whole blood sample clot at room temperature. The sample is then
centrifuged and the clot is removed, leaving a serum supernatant.
Working principle of centrifugation
A centrifuge is used to separate particles suspended in a liquid according to particle size and density,
viscosity of the medium, and rotor speed. Within a solution, gravitational force will cause particles of
higher density than the solvent to sink, and those less dense than the solvent to float to the top.
Centrifugation takes advantage of even minute differences in density to separate particles within a
solution. As the rotor spins around a central axis, it generates a centrifugal force acting to move particles
away from the axis of rotation. If the centrifugal force exceeds the buoyant forces of liquid media and the
frictional force created by the particle, the particles will sediment.
3.2.6 Anaerobic Jar
McIntosh and Fildes’ anaerobic jar is an instrument used in microbiology laboratory for the generation of
anaerobic conditions (anaerobiosis) to culture obligates anaerobes such as Clostridium spp. Anaerobiosis
obtained by McIntosh and Fildes’ anaerobic jar is one of the excellent and most widely used methods for
anaerobiosis but it requires costly special apparatus and a vacuum pump. Availability of gas supply is
another major drawback of this system. Currently, it is being replaced by a more convenient Gaspak
systems.
Working principle

20
McIntosh and Fildes’ anaerobic jar works on the principle of evacuation and replacement, where the air
inside the chamber is evacuated and replaced with mixture of gases (consisting of 5% CO2, 10% H2, and
85% N2).
It is practically impossible to evacuate all the air so some amount of oxygen will still be left behind. The
residual oxygen left behind is converted to water using spongy palladium or platinum catalyst. The
catalyst acts as a catalyzing agent causing slow combination of hydrogen and oxygen to form water.
Reduced methylene blue is generally used as an indicator (mixture of NaOH, methylene blue, and
glucose). It becomes colourless anaerobically but regains blue colour on exposure to oxygen.

3.2.7 Light Microscope

Instrument that produces enlarged images of small objects, allowing the observer an exceedingly close
view of minute structures at a scale convenient for examination and analysis.

Working principle
Light microscopes visualize an image by using a glass lens, and magnification is determined by, the
lens’s ability to bend light and focus it on the specimen, which forms an image. When a ray of light
passes through one medium into another, the ray bends at the interface causing refraction. The bending of
light is determined by the refractive index, which is a measure of how great a substance slows the speed
of light. The direction and magnitude of the bending of the light are determined by the refractive indexes
of the two mediums that form the interface.
A medium with a lower refractive index such as glass to air normally speeds up the light penetration and
makes light bend away from the normal and when light is passed through a medium with a greater
refractive index such as air to glass, it normally slows down and bends towards the normal,
perpendicularly to the surface.
If an object is put between these two mediums i.e between water and air, in this case, a prism, the prism
will bend the light at an angle. This is how the microscopic lenses work, they bend the light at an angle.
The lens (convex) on receiving the light rays, focuses the rays at a specific point known as the focal point
(F-point). The measure of distance from the centre of the lens and the focal point is known as the focal
length.

3.2.8 Biochemistry Analyser

A biochemistry analyser, also known as a clinical chemistry analyser, is used to measure metabolites
present in biological samples such as blood or urine. The study of these liquids makes it possible to
diagnose many diseases.
Working principle
You may recall the iodine-starch reaction experiment from a school science class. When you add iodine
solution to a starch solution, the solution will turn blue instantly. The existence of starch can be confirmed
in this way. In addition, if saliva is added to the coloured solution, the colour will disappear. This is
because the starch is dissolved by the function of an enzyme called amylase contained in saliva. An

21
automatic biochemical analyser utilizes this kind of reaction, and converts the quantity of a specific
substance in blood into an amount of colour change for measurement. The analysis method of measuring
the amount of colour change is called the colorimetric analysis method.

3.2.9 ELISA Reader Machine

An ELISA reader is an instrument that is used to read the fluorescent, chemiluminiscent, or chromogenic
response of the ELISA in a 96-well plate. The instrument is designed to fit standard 96-well plates, and
will provide quantitative information about the extent of the response for each individual well of the plate.
The ELISA reader is both a fluorimeter (able to input light at an excitation wavelength for the
fluororophore being used as the label, and to record the intensity of the emitted light at the emission
wavelength), and a spectrophotometer (able to record the intensity of light emitted from
chemiluminescent reactions and chromogenic reactions across a range of wavelengths.

Working principle
ELISA works on the principle that specific antibodies bind the target antigen and detect the presence and
quantity of antigens binding. In order to increase the sensitivity and precision of the assay, the plate must
be coated with antibodies with high affinity. ELISA can provide a useful measurement of antigen-
antibody concentration.

22
CHAPTER FOUR
4.0 GENERAL OVERVIEW.
The facility has the following units Media and sterilization, Bacteriology, Parasitology, Serology,
Biochemistry and the Pathology Section (Post Mortem)
4.1 MEDIA AND STERILIZATION
This unit entails machines that are used in autoclaving and preparation of media. They include.
Autoclaves, distiller, weighing balance, hot air oven among others.
Bio-risks in this unit include:
i. Slippery floors.
ii. Old cultures.
iii. Electricity,UV light.
iv. Sharp objects e.g. broken glass.
v. Chemicals.
vi. Hot surfaces.
vii. Operational equipment.
viii. Media and agars.
a. Media preparation.

 In this section, making and sterilization of media is done. Media prepared include: peptone water,
Simmons citrate agar, blood agar, Mac conkey agar, Glucose vi Phosphate, Tryptone Bile agar,
Hugh and Leifon’s medium(OF) medium
Types of Media
i. Basal media- supports growth of bacteria that do not need enrichment e.g. nutrient broth, nutrient
agar, and peptone water. Staphylococcus and Enterobactericeace grow in these media.
ii. Enriched media- are usually enriched by adding blood, serum or egg e.g. blood agar
iii. Selective media- favour the growth of a particular bacterium by inhibiting the growth of
undesired bacteria and allowing growth of desired bacteria e.g. Mac Conkey agar, Selenite broth.
iv. Differential/indicator media- contains an indicator where a particular microorganism causes
change in the indicator e.g. blood, neutral red, tellurite.
v. Transport media- used when specimen cannot be cultured soon after collection e.g. Cary Blair
medium, Stuart media.
vi. Storage media- used for storing the bacteria for a long period of time e.g. Egg saline medium,
chalk cooked meat broth.

23
• Blood agar.
Blood agar is a solid media used that is differential media in terms of production of haemolysis, and it is
an enriched media since it contains nutrients required to support a wide variety of organisms. We used
blood ager infusion agar. We dissolved 10gms of agar into 250mls in distilled water. The standard
proportion of preparation is dissolving 40gms of agar in 1000mls of distilled water.
• Mac conkey agar.
Mac conkey agar is a selective media that selects against gram positive bacteria since it contains bile salts
which inhibits growth of gram positive bacteria. It is also differential media since it contains an indicator
crystal violet which differentiates lactose fermentors from lactose fermentors.(lactose fermenters have
pink colonies). We dissolved 12.875gms in 250mls of distilled water. The standard proportion for
preparation is dissolving 51.5gms into 1000mls of distilled water. It was given a batch number 008.

 Simmons citrate agar.

Simmons citrate agar is a solid media used to differentiate gram negative bacteria on the basis of citrate
utilization. We dissolved 1.81gms of reagent to 50mls of distilled water. The standard proportion for
preparation is suspending 36.23gms in 1000mls of distilled water. It was given a batch number 001.

 Peptone water preparation.

Peptone water is a liquid media that is used to mainly to multiply bacteria for subcultures. The standard
proportion of peptone water is 15grms of reagent to 1000mls of distilled water therefore we dissolved
3.75grms in 250mls of distilled water then followed the standard operating procedures during preparation.
We then dispensed to 83 bijou bottles and gave a batch number 008.

 Glucose vi Phosphate
Glucose vi Phosphate is a liquid media which is prepared by adding peptone water, D-Glucose sugar,
Potassium Hydrogen Phosphate using the ratio 1:1:1 ie. 2.5 g of each in 500 mls of water.

 Tryptone Bile Agar

This media is used in the rapid detection and enumeration of E. coli. In the preparation you suspend 36.5
grams in 1000ml distilled water, sterilize by autoclaving at 121 0 C for 15 Minutes

b. Cleaning of special glassware.

Procedure.
i. Dirty glassware is received from bacteriology and other sections.
ii. They are entered into a register for records.

24
iii. They are then decontaminated by autoclaving at 121 degrees for 15 minutes.
iv. They are then soaked in tap water for 24 hours.
v. They are then washed and rinsed with distilled water.
vi. They are then wrapped using a paper foil.
vii. They are then sterilized by autoclaving at 121 degrees for 15 minutes.
viii. They are then dried in a hot air oven.
ix. They are then stored.
x. They are entered in a record book.

c. Collection, preparation of defibrinated blood from sheep.

Procedure.
i. Choose a healthy sheep then restrain it appropriately.
ii. Take a blood sample in a hepparised capillary tube then run a pcv (packed cell volume). This
sample is taken from the ear vein or coccigeal vein. The normal range of a sheep is 28-42. This should be
carried out for quality and control sensuality every fortnight.
iii. Sterilize the area around the neck.
iv. Shave the hair around the area of collection.
v. Locate the jugular vein.
vi. Using an 18 gauge needle and an adapter draw the blood to a conical flask containing glass beads.
vii. Swirl gently until the fibrin is formed.
viii. Store in a fridge until when need arises.
ix. Take a sample for quality control testing.

4.2 BACTERIOLOGY UNIT

In this unit, culturing of bacteria and other tests are carried out e.g. California mastitis test, somatic cell
count and quality control. Microscopy is also done. Samples brought to this unit include milk samples,
blood samples among other samples. This unit mainly carries out culturing and isolation of bacteria to
diagnose disease causative agents and prescription by carrying out Antibiotic susceptibility tests.

California mastitis test.

25
This is a milk test that is used to diagnose mastitis in milk. Its procedure is: Measure 2mls of the milk
samples from each quarter and place in a compartment then add equal amounts of reagent each quarter.
Observe for clots and any color changes. This test gives a rough estimate to the test as it cannot be used as
a confirmatory test.
Somatic cell count test.
This test gives the exact somatic cell count in a milk sample. This test gives a more particular result and
can be used as a confirmatory test apart from culture. Its procedure is: place 50mls of reagent to 50mls of
sample (it uses a lysis buffer) use a somatic cell counter cassette to take a portion of the mixture and place
in in a somatic cell counter machine.
SCC RANGES
0-200,000 ------------------ NEGATIVE
200,001-400,000------------ TRACE
400,001-1,200,000---------- 1
1,200,001-5,000,000-------- 2
+5,000,000--------------------- 3

A- < 10000 C- < 10000

B- < 10000 D- < 10000

Primary culture
Once a sample is received it is checked for correct labelling and recorded in a Laboratory internal form.
Primary cultures are made in blood agar and Mac CONKEY agar and incubated overnight under 35 0
Milk culture, use a sterile wire loop and make a primary inoculum by smearing. Streak is done and after
each streaking you sterilize the wire loop.
Primary cultures of tissues from the pathology nit, use a heated scalpel and burn the surface of the tissue
and then insert a sterile wire loop and streak in the medium.
Mac CONKEY Agar is used to differentiate lactose fermenters from non-lactose fermenters.
Blood Agar allows growth of both Fastidious and Non Fastidious bacteria.
Secondary Culture
Secondary Culture is done because of two main reasons: to obtain a pure culture and to multiply the
number of colonies. Secondary cultures are done on Blood Agar.
Microscopy.

26
This is done to describe bacteria and other organisms. Bacteria sample is taken from a culture then they
are gram stained then observed under a microscope. The cultures are reported to be either gram positive
or gram negative strains of either cocci or rods.

Culture and isolation of bacteria.

Cultures of bacteria on blood agar
Differentiates organisms by their ability to produce hemolysis.
Hemolysis on B/A
Blood contains inhibitors for certain bacteria such as neisseria and hemophylus genera. So the agar must
be heated to inactivate this genera, and to release essential growth factors. Heating converts it to
chocolate agar.
Types of hemolysis produced are; alpha, beta and gamma hemolysis.
Description of cultures
Described based on size, color, texture/consistency, and the type of hemolysis produced.
Mac CONKEY differentiates between lactose fermenters and non-lactose fermenters.

Preparation of slide
a. Take a glass slide and put a drop of normal saline
b. Pick a single colony
c. Make a smear on the slide
d. Air dry
e. Gram staining
Gram stain
Procedure;
i. Air dry smear and apply heat fixation for two minutes
ii. Crystal violet is filtered onto the slide and decanted after two minutes
iii. Lugol`s solution is applied then decanted
iv. Decolorize with acetone for a split second and rinse with tap water immediately
v. Apply carbol fuschin for 2 minutes and wash off.
vi. Incubate to dry

27
vii. Proceed to microscopy

Biochemical Tests
Setting of Biochemicals depends on the gram stain produced.
Gram Positive cocci are cultured in Manitol Salt Agar medium, gram negative rods are subjected to the
IMVICUM tests(Indole, Methyl red, voges Proskeur, Indole, Citrate, Urease, Mannitol) and the gram
positive rods are cultured in peptone water and Glucose vi Phosphate.
Reporting of Biochemical tests
Cultures of Biochemicals will have a change in colour, gas production depending on each medium.
Biochemical tests are performed in order to identify the bacteria depending on their characteristic
produced in each medium.
These tests involve growing the organism in various carbohydrate containing media (sugars) and
observing which sugars are altered by the organism to produce acid. Other tests detect end products of
bacterial growth such as indole and hydrogen sulphide while others test the organism for enzymatic
activity such as oxidase , catalase e.t.c.

Antibiotic sensitivity/susceptibility
This test is done to help choose the antibiotic that will be most effective against the specific types of
bacteria or fungus infecting an individual person or animal. It can guide the doctor the drug of choice and
dosage to treat infections. Disc diffusion method is used to determine antibiotic sensitivity where discs
impregnated with various antibiotics are placed on the surface of an agar plate that has been inoculated
with the organism isolated from the sample. The antibiotic diffuses outward from the disk over a standard
incubation time and the diameter of the zone of inhibition is measured. The size of this zone is compared
with standards to determine the sensitivity of the organism to the drug.

4.3 PARASITOLOGY UNIT

Protozoology Section
In this unit, tests for protozoan (haemoparasites) parasites are done. Diagnosis is also done for protozoan
diseases e.g. East coast fever, Anaplasmosis, Babesiosis and Theileriosis.
TYPES OF PROTOZOOLOGICAL SAMPLES
For babesiosis samples are blood smears or blood gland smear.
For anaplasmosis samples are blood smears or blood gland smear.

28
For east coast fever samples are blood gland smear.
For trypanosomiasis samples are blood smear blood gland smears and EDTA.
Cowdriosis/ heart water samples are brain squash smears.

Tests done in this unit:

Packed cell volume
This test is done to measure the proportion of erythrocyte cell count in a blood sample in.
Procedure: obtain a blood sample intended to be tested using a heparinised capillary tube, fill the capillary
tube to three quarter, wipe excess blood on the tube, lock the tube on one side using plasticine then place
the tubes in a centrifuge. Centrifuge at 1200 revolutions per minute.
Read the results using a haematocrit reader.
The normal range of Packed Cell Volume is 24%-46% in cattles and slightly above in calves.

Making of blood and gland smears.

Blood smears are made in three forms:

 Thick blood smears are made so as to observe parasites that are extracellular e.g. trypanasomiasis
parasites.
 Thin blood smears are made to observe individual cells and intracellular parasites.
 Impression smears are made during post mortem.

Procedure of making a blood smear:

Put a drop of blood on one end of a slide, using another slide make one streak that is smooth advancing
forward ensuring you obtain a head and a tail. Thick blood smears are obtained on the head while thin are
obtained from the tail. They are then stained using giemsa stain and then observed under a microscope.
The slide is first fixed in ethanol to fix it for a few minutes.
Performing a GIEMSA stain
Make a GIEMSA stain reagent by mixing with buffered water in a ratio of 1:10 (1 part of GIEMSA in 9
parts of buffered water). Then submerge the slide in the GIEMSA stain for 30 minutes.
Microscopy
Observe the slide under a microscope at power ×100 with oil immersion.

29
Anaplasma marginale
Others observed include:
Babesia bovis
Koch’s blue bodies.

Helminthiology section
In this unit they mainly deal with helminthes. They include trematodes, nematodes and cestodes. Samples
brought to this unit are mainly fecal samples and in few cases stomach contents from the pathology
section.
Tests done include:
i. Floatation technique which lets eggs with a low refractive index to float these eggs includes
strongyle and cestode eggs. Procedure:

 Calibrate a beam balance. We used a triple beam balance.

 Weigh 3gms of fecal sample.
 Emulsify with 30mls of tap water.
 Crush using a homogenizer.
 Measure 57mls of saturated salt solution (sodium sulphate, magnesium sulphate, or sodium
chloride.)
 Pour the sample through a tea strainer to the saturated salt solution.
 Use a dropper to stir the sample vigorously to send the eggs into a suspension and quickly
withdraw a sample and fill both compartments of a Mac master slide.
 Mount the Mac Master’s slide on a microscope and give it 5minutesfor the eggs to rise up.
 Focus and observe at power ×10.
 Count eggs from all chambers and those within columns of the chambers and those that fall on
the lines of the chambers excluding eggs that fall on the marginal lines of the chambers.
Calculate using the factor: Average of (Chamber 1+ Chamber 2) multiply by 50 eggs/gram.

4.4 PATHOLOGY UNIT (POST MORTEM)

The pathology section constitutes of the reception and post mortem. At the reception, different clients
with different enquiries are received and attended to by the receptionist and the pathologist who is on
duty. Samples are submitted by farmers, veterinary field officers and private practitioners.
Once the sample is received, the officer on duty at the reception takes the details of the animal from
which the sample was collected and captures the disease history, species of the animal, age, sex and
duration of illness and the test requested. Details of the owner are also collected including the name,
address, phone number, county, sub-county, location and the village are also recorded for ease of follow –
ups and facilitation of further investigations through farm-visits and active surveillance.

30
The person who submits the samples is required to pay some money depending on the test requested. The
sample is then labelled with an identification number according to species and recorded in the sample
register. Recording is captured manually and then stored electronically (LIMS) for data management and
as required by ISO.
Depending on the test requested, an inter-lab form is then filled capturing the sample details, laboratory
identity number (case number), date, and the laboratory to which it is to be sent. The sample is then sent
to the particular lab accompanied with the inter-lab form.
Samples that are received at the reception includes whole blood (EDTA), faeces, serum, urine, carcasses,
live animals, vaginal swabs, nasal swabs, organs like livers, kidneys and blood/gland smears (can also be
impression smears).
Animal species received include bovines (cows), ovines (sheep), equine (horse), avians (birds eg hens),
canines (dogs), porcines (pigs), felines (cats) and miscellaneous e.g. camels.
Animal carcasses received for postmortems should not be opened until microscopy is done to ascertain
absence of Bacillus anthracis, a gram positive bacilli that has terminal spores and to prevent the release of
bacillus spores into the environment. Microscopy also rules out other haemoparasites e.g. Babesia spp,
Anaplasma spp, Theileria parva etc which may have caused the death of the animal. This is done by
preparing blood smears and staining under hematological techniques. The smear should be prepared such
that the head of the smear is thick and the tail thin. The thick portion is used to scan the blood while the
thin end is used for final identification. Staining of the blood smears applies the use of stains like Giemsa,
Rowmanosky stains or Wrights stain. These stains allow for the detection of white blood cells, red blood
cells and platelet abnormalities.
Once microscopy rules out presence of Bacillus anthracis, the pathologist on duty opens the carcass and
records any pathological change observed and can proceeds to collecting organs as samples for further
diagnosis in the laboratory. The carcass is then disposed into a Bio pit and the pit is well closed.

Cases handled include:

Bovine:
1) A 6 months old calf.
Clinical signs: dyspnoea, lacrimation, enlarged sub pleural lymph nodes,
Post mortem findings: sunken eyes, edema in the lungs, emphysema, fluids in the bronchi, hemorrhage in
the anterior side of trachea and the kidney, dry ruminal ingesta.
Diagnosis: East coast fever.

2) A adult female Friesian cow.

Clinical sins: restlessness, recumbency, off feeds, labored defecation.
Post mortem finding: hydro peritoneum (whole blood), fluids in the abomasums, fluids in the rumen,
enlarged heart, torsion of the abomasum on the short axis.

31
Diagnosis: torsion of abomasum.

Avian:
1. 2 hens.
Clinical signs: ruffled feathers (fever), reddened head.
Post mortem finding: hard pealing skin (dehydration), clear mucoid in the trachea, slight hemorrhage in
the trachea, ovarian tissue, spleen, adhesions between liver and body wall, adhesion between gizzard and
body wall, congestion in the liver, necrosis in the liver, cloudy air sacs, cheesy and creamy mass in the
coelom, enlarged kidney filled with urate and a reddened ovarian tissue.
Diagnosis: collibacillosis.
2. 14 hens (7 live, 7 dead)
Clinical signs: ruffled feathers, depression.
Post mortem findings: congestion and hemorrhages in the eyes, brisk muscles, heart, spleen, in the bursa
of fibricus, proventriculus, enlarged urate filled kidneys, bronze color discoloration of liver, edematous
unregressed bursa of fibricus .
Diagnosis: infectious bursa disease.
Samples taken: heart, spleen, liver, portions of trachea and intestines, bursa of fibricus.

4.5 SEROLOGY SECTION

The serology section deals with diagnosis of infectious diseases through tests involving antibody antigen
reactions tests like Enzyme Linked Immuno-sorbent Assay (ELISA), Complement Fixation Test (CFT),
Milk Ring Test (MRT) and Rose Bengal Plate Test (RBPT).
Diseases tested include Contagious Bovine Pleuro-Pneumonia (CBPP), Contagious Caprine Pleuro-
Pneumonia (CCPP), Brucellosis and Johnes disease. Serum samples submitted are used for diagnosis of
CBPP, Brucellosis and Johnes disease while milk sample are used for diagnosis of Brucellosis. Rose
Bengal Plate Test and Milk Ring Test are screening tests for Brucellosis.

Indirect ELISA Test for Brucella abortus

Antibody can be detected by indirect ELISA. In this technique, antigen is coated on the microtiter well.
Serum or some other sample containing primary antibody is added to the microtiter well and allowed to
react with the coated antigen. Any free antibody is washed away and the bound antibody to the antigen is
detected by adding an enzyme conjugated secondary antibody that binds to the primary antibody.

32
Unbound secondary antibody is then washed away and a specific substrate for the enzyme is added.
Enzyme hydrolyses the substrate to form a coloured product. The amount of coloured end product is
measured by spectrophotometry plate readers that can measure the absorbance of all the wells of a 96 well
plate.

Test Principle
The test principle of ELISA is based on antibody-antigen reaction. The microtiter wells are coated with
the capture antigen that is specific to the antibody in the test. The test sample is added to the antigen to
form an antigen-antibody complex. The unbound antibody is washed a buffer. A conjugate is then added
containing an enzyme conjugated to a secondary specific to target primary antibody. Enzyme commonly
used is horseradish peroxidase. The unbound conjugate is washed away using a buffer after which a
substrate is added. The substrate is catalysed into a coloured product whose optical density is read using a
spectrophotometer.
Requirements
Micro title plates
Micro pipette
Coating reagents
Test kit
Buffer

Procedure
i. Reagents are equilibrated to room temperature (18-25°C) before use. Each strip is given a
number.
ii. Samples are added with the negative and positive control serum provided being used for both
serum and milk testing.
iii. 100µl of sample dilution buffer is added to each well that will be used for serum samples and
serum controls.
iv. 4µl of positive control serum and negative control serum is added to selected wells coated with
Brucella abortus antigen. For confirmation purposes, it is recommended to run the control sera in
duplicates.
v. 4µl of serum samples is added to selected wells coated with Brucella abortus antigen. The
samples can be tested in duplicates.
vi. The plate is shaken thoroughly and sealed with a strip and incubated at 37° C for one hour.

33
vii. The plates are rinsed three times with PBS Tween buffer, filling up the wells at each rinse,
emptying the plate and tapping hard to remove all remains of fluid.
viii. 100µl of HRP conjugate is added to each well, sealing the plate and incubating at 37°C for 1
hour.
ix. Step No. 4 is repeated.
x. 100µl of substrate solution is added to each well and incubated for 10min at room temperature,
with timing began after the first well is filled.
xi. The reaction is stopped by adding 50µl of stop solution to each well and mixed thoroughly. The
stop solution is added in the same order as the substrate solution in step 7.
xii. The optical density of controls and samples is measured at 450nm in a micro plate photometer
within 15min after adding the stop solution to prevent fluctuation.

Results and Interpretation

Samples are exposed to non-infectious Rubella antigen coated wells on micro titer plates. Rubella
antibodies if present in the test sample bind to the antigens in the well. HRP conjugate added
subsequently forms a complex with Brucella antibodies. Unbound material is removed by rinsing before
the addition of a substrate solution. A blue colour develops which is due to the conversion of the substrate
by the conjugate. A positive result is indicated by development of a blue colour. A reaction is stopped by
the addition of the stop solution, the colour changes to yellow.
Rose Bengal Plate Test (RBPT)
RBPT is commonly used testing sera collected from cattle suffering from Brucella abortus. It is an
agglutination test carried out on white plate/tile. The Rose Bengal reagent is stained with a pink colour.
Equal amount of staining and antibody in the test sample placed on a white plate which is non-absorbent.
Brucella specific antigen will bind to the antibody in the test serum forming agglutination. A positive
presents as agglutination on the plate while a negative shows no clumps.
Requirements
White tile
B. abortus antigen
Micro pipette
Procedure
i. Test Serum (0.03 ml) is mixed with an equal volume of antigen on a white tile or enamel plate to
produce a zone approximately 2 cm in diameter.
ii. The mixture is agitated gently for four minutes at ambient temperature, and then observed for
agglutination.
iii. Any visible reaction is considered to be positive.

34
Results and interpretation
When an animal is infected with Brucella abortus, the immune system of the animal produces antibodies
against B. abortus antigen. Specific antigen bind to a specific antibody forming agglutination complex,
when clumps are formed is a good indicator that the animal is infected.
The test was negative since there was no clumps formed after agitating the mixture.
Rapid Antigen Test for Newcastle Disease in Avian
Newcastle disease is a viral infection with specific IgG antibodies in chicken. The disease is caused by
virus called Paramyxo virus. NDV is complicated because different isolates and strains of the virus may
induce variations in the severity of the disease even in a given host.
Requirements
Rapid test kits.
Phosphate buffer.
Disposable droppers.
Croacal swabs preserved in peptone water.
Procedure
i. Add two drop of the solution (croacal swab + peptone water) into the kit.
ii. Add phosphate buffer solution and let the kit rest for three minutes.
Results and interpretation
A colour band will appear in the left section of the window as the control band. This shows that the kit is
working properly. The right section of the window indicates test results. This band is the test band. If the
control band does not appear in the result windows the test is considered invalid. Only one band appeared
control band which indicated that the test was negative for Newcastle Disease Virus (NDV).
Rabies Test
Principle
The Antigen Rapid Rabies Ag Test Kit is a chromatography immunoassay for the qualitative detection of
Rabies virus antigen in canine, bovine, raccoon dog’s secretion of saliva and brain homogenates. The kit
has a letter ‘T’ and ‘C’ as test line and control line on the device. The specially selected Rabies virus
antibodies are used in test band as both capture and detector materials.
Requirements
Antigen Rapid Test Kit
Sample collection swabs.
Disposable droppers.
Buffer solution.

35
4.6 BIOCHEMISTRY SECTION
It is a science of using the chemical analysis of body fluids to obtain information about clininical
conditions of the body.
Blood fluids used include: serum/plasma, cerebrospinal fluid, saliva etc., sweat
In many diseases there are changes in the chemical composition of fluids tests are done to determine the
differences between sick and normal animals. Changes can be qualitative/ quantitative therefore used as a
tool in clinical Biochemistry. This provides diagnosis, prognostication, and treatments of a disease also do
preventions. Substances measured are:
a. Substances measured in serum include; glucose, total protein, individual protein, cholesterol,
hormones, vitamins using a Biochemistry analyzer.
b. Metabolites in circulation, non functioning waste products that are in the process of being cleared
in the body e.g. urea, billirubin, creatinin, ammonium, etc
c. Substances released in cells as a result of damage e.g. abnormal permeability of cell wall,
abnormal cell divisions, enzymes etc
d. Drugs and toxic substances e.g. ammonium therapeutic rugs. It is easier to detect a malfunction
rather to determine a cause e.g. could be a destruction of cells, genetic defect, insufficient blood supply,
malignancy, and defect in cellular recognition of certain things.
Demonstration of the working of the analysis of; glucose, total protein, individual protein, cholesterol,
hormones, vitamins was done using Biochemistry analyzer.

4.7 FIELD ACTIVITIES

i. At the Slaughter house
We visited the local slaughter house as one major duties of the facility is inspection and surveillance for
livestock diseases even in the animal products and by products like meat, skin etc.
Meat Inspection
Procedure of slaughter. Mainly divided into 2 stages.

 Antemoterm- animals are brought and held at a holding yard and let to rest for 24 hours while
they are being inspected whether they are healthy and are fit for consumption. At this point you
examine for bruises, emaciation, lumpy skin disease, foot and mouth disease, and swelling of
lymph nodes.
 Lie ridge- at this point examination is done checking for temperature, pulse rate, palpate lymph
node, mouthing- incase of foot and mouth disease and then check the mucus membranes.

36
  Stunning box-they use captive bolt pistol to stun.
 Post mortem- killing floor- the animal is severed to drain blood.
 Hoisting-the animal is then hoisted upside down for proper bleeding.
 Flaying- undress the animal.
 Evisceration- removal of all internal organs.
 Splitting- the carcass is split into two thereafter inspected
 Inspection- inspect main organs:
 Liver – check for flukes, cysts, abscesses and other worms.
 Spleen-check for abscesses, swellings and consistency.
 Heart- in bovine check for cystcercus bovis in all the sceptaes and the heart muscle.
 Kidneys- check for consistency.
 Lungs- palpate for swellings and check for necrosis and parasites.
 Muscles- check for cystcercus bovis in bovine muscles.
 Head-check for infections.
 Lymph nodes- slice them to check for infections and abnormality.

 Reasons for inspection.

 To examine for various zoonotic parasites e.g. taenia parasites, notifiable diseases e.g. anthrax,
brucellosis, non notifiable diseases e.g. contagious caprine pleuro pneumonia, CBPP, fasciolosis,
foot and mouth disease, among others
 To check for bone and meat diseases.
 Check for food poisoning in meat e.g. streptococcus, salmonellas, botulism, e.t.c
 To ensure safety for consumers.

ii. At Wambugu Farm-Field Day

Attended field activity at Wambugu farm located in Nyeri County. The event is held each year in
collaboration with the ministry of Agriculture.
We engaged farmers on how to collect samples for proper analysis of the test sample.
The event objectives is to enhance capacity building amongst the farmers and to self-aware them on
where to get laboratory tests done for their livestock.

37
CHAPTER 5
5.1 DISCUSSION
SKILL, TECHNIQUES AND LESSONS LEARNT AND THEIR RELEVANCE
During this training period, I have gained a lot of experience, knowledge and exposure. There were many
changes from the point of learning environments and discussion among colleagues, which has helped me
improve in skills like interpersonal relations, listening, presentation skills and acting freely around people.
I have learnt many lessons which includes; how to operate in a Biosafety cabinet, how to prepare different
media, how to operate the autoclave and much of importance how to produce quality work in the field of
Bacteriology. Other skills learned include how to perform different post mortems of different species.

5.2 CONTRIBUTIONS MADE TO THE INSTITUTION

During the attachment period I was able to deliver the following:
Routine cleaning and disinfection of laboratory and media section.
Routine laboratory work: diagnostic, analyzing of results and entering of laboratory records in the
Laboratory Information Management System.
Took part in a field activity to sensitize farmers on the methods of specimen collection and safe
transportation to the laboratory for diagnostic purposes.
Took part in a research project and collecting of data for the same.

5.3 PROBLEMS ENCOUNTERED

Serological tests are very expensive and it was thus difficult during the attachment period to carry out
tests on few submitted samples until they reached a threshold that allowed the opening and use of
reagents in the test kits.
Activities in the in the Biochemistry lab could not be carried out as the facility does not receive enough
specimens, since the reagents are expensive the management find it uneconomical thus we could not learn
practically but just through demonstration.

38
Large number of students on attachment in the facility hindered effective interaction of the technical staff
with the students due to congestion on benches and limited equipment and reagents.
There was less learning time to be attached in the institution as it would allow as for two months.
The institution operating on a referral system with Kabete Central Veterinary Laboratory we were
disadvantaged with cases of viral infection diagnosis being referred to Kabete.

5.4 RECOMMENDATION
More awareness to be made in the mandate region on the availability of Biochemistry and Serological
tests being performed in the laboratory so as to these section are able to run economically.
More funding should be channeled towards the institution to facilitate procurement and purchase of
equipment and reagents required for sample processing. In this way, farmers and other stakeholders will
not have to wait for results for longer times as more samples are waited to be submitted for these tests to
be carried out.
In the future, the institution should consider admitting fewer numbers of students as this promotes
effective interaction between the students and technical staff.
In the future the institution should give students more learning time i.e. three months as indicated in the
school letter.
More awareness needs to be created on the importance of such a facility and its capacity to carry out
diagnosis, surveillance, research, management and control of animal pests and diseases and regulatory
management and quality assurance of animals, animal products, by-products and animal
health/production inputs.

5.5 CONCLUSION
The attachment was a great opportunity to gain more experience and meet new friends. This enabled us to
interact and increase our knowledge on different areas. The field practices helped us gain more experience
on the theoretical work we had learnt at school. The staff was more than willing to help us throughout the
laboratory work. There were enough resources to ensure that we were well equipped with everything that
we need to our work. The doctors were of great help in ensuring that we were taught and acquired
knowledge that we needed. The facility has done more than expected to ensure smooth undertakings
during this period.

39
5.6 REFERENCE

40