1. health and fitness
2. disease
3. infertility

PRACTICAL CLASS MANUAL
VETERINARY PHYSIOLOGY-III (VPB-221)
(Endocrinology, Male & Female Reproduction, Lactation,
Growth and Environment Physiology)
Compiled by:
Dr. R. Kumar, Prof. & Head
Dr. K.B. Sharma, Prof.
Dr. M.S.Verma, Asstt. Prof.
DEPARTMENT OF VETERINARY PHYSIOLOGY & BIOCHEMISTRY
DR. G.C.NEGI COLLEGE OF VETERINARY & ANIMAL SCIENCES
HIMACHAL PRADESH KRISHI VISHAVIDYALAYA
PALAMPUR (H.P.) 176062
Third Edition (Reprint)
2014
PRACTICAL CLASS MANUAL
VETERINARY PHYSIOLOGY-III (VPB-221)
(Endocrinology, Male & Female Reproduction, Lactation,
Growth and Environment Physiology)
Name of Student ________________________
Admission No.
________________________
Compiled by :
Dr. R. Kumar, Prof& Head
Dr. K.B. Sharma, Professor
Dr. M.S.Verma, Asstt. Prof.
DEPARTMENT OF VETERINARY PHYSIOLOGY& BIOCHEMISTRY
DR. G.C.NEGI COLLEGE OF VETERINARY & ANIMAL SCIENCES
HIMACHAL PRADESH KRISHI VISHAVIDYALAYA
PALAMPUR (H.P.) 176062
Third Edition (Reprint)
2014
1
Foreword
I am very pleased to learn that teachers of Department of Veterinary Physiology have
brought out the Second Edition of “Practical Class Manual on Endocrinology and
Reproduction”. The faculty of Department of Veterinary Physiology have already prepared
Practical class manuals on Cardiovascular & Respiratory System, Digestive & Excretory System
and Growth and environmental physiology for all the undergraduate courses as per VCI course
curriculum, which will be used by the students of this college. This manual will definitely be
useful in understanding the intricacies of the laboratory exercises.
I am sure the Second edition of Practical Class Manual on Endocrinology and
Reproduction with many new additions will help the students
to enhance their skill and
understanding of these exercises.
I wish to congratulate the authors of Manuals for this great effort.
Dean,
Dr. G C Negi College of Veterinary & Animal Sciences,
CSK HPKV, Palampur
2
PREFACE
There is a general feeling among the Veterinary academia that laboratory exercises and
practicals have not been getting due in the curriculum. This feeling has been gaining ground
especially in the land grant system of education which is currently in vogue in most of the
Agricultural Universities of the country.
It is felt that every laboratory exercise or topic can not be explained, shown or got done
by individual student in a short duration (two hours) of laboratory class. There was, therefore, a
long felt need to bring out system – wise “Practical Class Manuals” which could serve as a
handy laboratory aid to the individual student.
Department of Veterinary Physiology have already prepared Practical class manuals on
Cardiovascular & Respiratory System, Digestive & Excretory System and Growth and
environmental physiology for all the undergraduate courses as per VCI course curriculum. The
present Second edition of manual on Endocrinology and Reproduction has many additions and
some of the exercised has been updated. With the manual handy with each student, he would be
expected to come too the laboratory class well prepared and get to the business of conducting
and experiment right away.
Besides explaining the individual exercise on different systems, the manual contains
appendices of normal values and spare pages for write-ups by the students.
The author express gratitude to all those to helped us in various ways and invite
suggestions from readers to improve the content and style of the manual.
R. Kumar
K.B. Sharma
M.S.Verma
3
TABLE OF CONTENTS
Sr.
Name of Experiment
No.
Page
No.
1
An introduction: male and female reproductive organs
of domestic animals.
5
2
Phases of estrus cycle in domestic animals
10
3
Detection of oestrus in farm animals.
12
4
Semen and its components.
14
5
Morphological evaluation of semen.
16
6
Microscopic evaluation of semen.
18
7
Determination of live and dead sperm count.
20
8
Common abnormalities of spermatozoa.
23
9
Measurement of hormone concentration in body fluids.
26
10
Effect of insulin on carbohydrate metabolism.
29
11
Growth measurement in different species of domestic
30
Date
Instructor
Signature
animals
12
13
14
15
16
17
18
Instruments and Equipment’s used for measurement of
physical environment/Climate
Basic Requirement of a meteorological station/
observatory
Climatic Change related to environmental physiology
and the effect and relationship of environment/
weather on animals
To record various physiological parameters and study
the effect of extreme weather on animal.
Annexure –I Comparative anatomy
36
Annexure –II Duration of oestrous cycle and its phases
in different species of animals
Annexure –III Seminal characteristics in different
species of animals.
54
42
45
46
53
55
4
Exercise -1 Introduction to male and female reproductive organs
of domestic animals.
Sexual differentiation of male/ female reproductive organs
The sexual differentiation, like the differentiation of other systems proceeds in
consecutive steps. Chromosomal sex is determined at the time of fertilization and establishment
of gonadal sex is followed by differentiation of the Mullerian and Wolfian duct systems into
female and male accessory genitalia respectively. The final step in the establishment of psychic
sex is with the characteristic male or female sexual behaviour.
Homologies of Male and Female Reproductive system
Male
Female
Indifferent internal gonad
Testis, Rete testis
Ovary, Rete Ovary
Mesonephric tubules
Vas efferens, Paradidymis, Epoophoron, Paraphoro
Vas Aberreans.
Mesonephric Duct
Epididymis, Vas deferens, Gartner’s duct
Ejaculatory duct, Appendix
of epididymis
Mullerian Duct
Appendix of testis, Prostatic Appendages of ovary, ovary
utricle ( Uterus Muscularis)
fimbria of oviduct, uterus,
vagina
(part
of
external
genitalia)
Urinogenital Sinus
Prostatic membranous and Urethra,
cavernous
vestibule,
vagina
urethra, (part of ), vestibular gland,
Bulbourethral gland, prostate Para urethral gland.
gland
Genital Tubercle
Glans Penis, Corpus Penis
Glans Clitoris, Corpus C
Urethral folds
Raphe of scrotum and Penis
Labia Minora
Labioscrotal Swelling
Scrotum
Labia majora
5
(I)
Female reproductive organs:
-
Ovary
-
Tubular genital duct/ fallopian tube
-
Uterus
-
Cervix
-
Vagina
-
Vulva
The organs of reproduction in female are ;
1. Ovary :
The ovaries serve a double function similar to the testis. Both organs have a gametogenic
and an endocrine function. This dual role is complimentary, since gametogenesis requires certain
changes in the reproductive tract to complete reproduction. Response of reproductive tract is
caused by the gonadal hormones. Ovaries are paired organs and there is some difference in
functional capacity of the two ovaries in various species of animals. Ovaries are located in
abdominal cavities. Gross shape of ovaries varies according to species and stage of estrus cycle.
The polytoccous animals ( Sow, Bitch and Cat ) have several follicles or corpus luteum, giving
appearance of cluster of grapes, while in monotoccous ( Cow, Mare, Ewe) have more ovoid
shaped ovaries unless a graffian follicle of corpus luteum is present. The graffian follicles are of
following types.
I. Primordial / Primary Follicles
II. Growing / Secondary Follicles
III. Tertiary / Graffian follicles
IV. Atretic Follicles
2. Tubular genital tract:
Tubular genital tract of female serves as the transportation route for sperm in uterine tube
for fertilization. Uterine tube also called as oviduct or fallopian tube are paired convoluted
tubules that reaches from ovaries to uterus. The ovarian end is funnel shaped that helps in
collecting ova at the time of ovulation. Middle portion of uterine tube is called as ampulla and
next to uterus is isthmus. There is no demarcation at point of junction of uterine tube and uterus.
3. Uterine Tube:
It is a tortuous organ of small diameter in all domestic species. Lumen of uterine tube is
honey combed in appearance. Epithelial lining of the lumen of uterine tube is also termed as
“tunnel of love” for transporting ova, sperm and zygotes. In domestic animals uterus consist of
two horns ( cornua ) and a body. Development of longer horns in certain species (bitch & sow)
is related to delivery of litters. Uterus is supported by broad ligaments of uterus, which is subject
6
to considerable stretching during pregnancy. During non-pregnancy the uterus is held in the
dorsal pelvic area. Uterus is composed of three distinct layers:
I. Serous membrane
II. Myometrium
III. Endometrium
- it is extension of peritoneum.
- it consists of three muscle layers.
- it comprises of epithelial lining of lumen glands and
connective tissue.
4. Cervix :Cervix is the doorway for uterus, a physiological barrier separating the external
environment from internal environment of the animal. Beyond the cervix lies uterus, a perfect
incubator for zygotes and microorganisms. Outside cervix lies vagina. Cervix is a thick walled
sphincter like organ. It has a thick muscular wall capable of contracting to close the passageway
or relaxing to accommodate movement of semen during estrus phase. Lumen of cervix is
tortuous, since it is composed of many folds called annular rings. Cervix has tall columnar
epithelial cells mixed with goblet cells, which have important secretary functions. Goblet cells
secretes mucus depending upon stage of estrous. During estrus the cervix is hyperemic.
5. Vagina :Vagina serves as passage for the foetus outward at parturition and for semen inward
following copulation. Exterior limits of vagina marks the confluence of urinary tract with
reproductive tract. Epithelial lining of vagina undergoes cyclic changes under influence of
ovarian hormones.
6. External Genitalia / Vulva :
It consists of labia majora, labia minora and clitoris.
7
Male reproductive organs: The organs of reproduction in males are :
-
Testis
Rete testis
Epididymis
Vas – deferens
Seminal vesicles
Cowper’s gland
Ampullae
Prostate Gland
Urethra
Penis ( in mammals )
Ejaculatory duct ( in birds)
1. Testis :
It descends from abdominal cavity into scrotal sac, much before puberty, except in
elephants and whales. In seasonal breeders testis ascends back into inguinal canal during nonbreeding season.
Each of the testis contains convoluted seminiferous tubules, (diameter – 0.2 mm, length
– 50 to 100 mm) which communicates to rete testis then through epididymis and vas- deferens to
the urethra. Sertoli cells are large basally situated having prominent nucleoli and provides
nourishment to the developing sperms and probably also produces androgens and estrogens.
Sertoli cells are attached to the basement membrane and have numerous long processes which
may contact all other cells with in seminiferous tubules. They are believed to serve as nurse cells
for the developing spermatids.
Blood supply to the scrotal testis is also peculiar. Testicular artery becomes convoluted
just above the testis and is largely surrounded by veins taking out blood from testis. Veins from
a pampiniform plexus. At this site heat is exchanged with counter – current mechanism. Heat
exchange process is important since the spermatogenesis occurs at the lower temperature than
the body temperature ( 32-35 ºC), so the arterial blood reaching the testis is cooled down by
venous blood.
2. Rete Testis : Rete testis fluid is mainly contributed by sertoli cells. The blood plasma crosses the
blood – testis barrier and some constituents’ passes into rete – testis fluid. This fluid provides the
liquid medium for the movement of spermatozoa to epididymis. As most of the fluid of retetestis are reabsorbed in epididymis. Hence the spermatozoa concentration is lower than
ejaculated semen in rete testis. Fluid does not contain glucose, but is rich in inositol, which is
energy source for spermatozoa. From here sperm takes 2-3 hours to reach epididymis.
8
3. Epididymis :
It is well developed in mammals with head, tail and body but poorly developed in
birds. It’s highly tortuous and total length is about 120 feet in bull and 180 feet in boar. Four
basic functions of epididymis are:
a. Transport b. Maturation c. Absorption & Secretion d. Storage of sperms.
4. Vas –deferens :
It is a muscular duct that transports spermatozoa from epididymis to urethra by peristaltic
movement during ejaculation, no secretions in this region; end of vas deferens is dilated to form
ampulla, which contains lot of gland and is more prominent in stallions.
5. Accessory sex glands :
a. Seminal vesicle
b. Prostate gland
c. Cowper’s gland
a. Seminal vesicle are paired glands and located lateral to ampulla. It is absent in dogs and
fowl. Secretion is highly viscous and milky. Secretion of seminal vesicles contributes
50% of the semen volume. In bull its yellowish due to riboflavin content.
b. Prostate gland is absent in fowl. Its secretion is little in quantity in most of the species
except in dogs where it contributes most of the seminal plasma.
c. Cowper’s gland/Bulbourethral gland: In boar its secretion is thick, white and rubber like
consistency and this causes coagulation of semen in boar. In other species its secretions
helps in flushing of urethra before ejaculation.
6. Ejaculation:
Ejaculation is the process of discharging semen out of male reproductive tract.
During ejaculation semen is not allowed to enter in urinary bladder due to the closure of
sphincter of urinary bladder.
9
Exercise II : Phases of estrus cycle in domestic animals.
Oestrus cycle: The oestrus cycle is a cycle of endocrine events associated with
morphological changes in the cows’ reproductive tract accompanied with certain behavioural
traits.
The reproductive cycle of adult female mammals results from fluctuations in the endocrine
activities of the anterior pituitary gland and the ovaries. The pituitary gonadotropins regulate
both the germ and non-germ tissues of the gonad, while the ovarian hormones control the
functional state of the reproductive tract and external genitalia. The estrus cycle is regulated by
pituitary- gonad axis with the release of FH/LH determining the cyclical fluctuations of estrogen
and progesterone which in turn affect the vaginal epithelium and uterus. Vaginal histology
changes during the cycle as a function of circulatory levels of the ovarian hormones. There are
four stages in the estrus cycle of the domestic animals(i)
Proestrus, (ii) Estrus (iii) Metestrus and (iv) Diestrus.
Proestrus ( Day 17-20)



Regression of the corpus luteum of the previous cycle and drop in the progesterone
concentrations in circulation.
The dominant follicle is selected and grows producing increasing amounts of oestradiol.
In late proestrus, the influence of oestrogens on the reproductive tract and behaviour of
the cow can be observed.
Oestrus (Traditionally considered as day 0 of a new cycle)






Oestrus is also called heat and lasts on average 4-24 hours.
During oestrus the cow/heifer is receptive to a bull and stands for mating (standing heat).
The dominant follicle reaches its maximum growth, matures and ovulates.
Ovulation is induced by high LH concentrations (preovulatory LH surge).
Ovulation takes place approximately 14h after the onset of oestrus and after the
behavioural signs of oestrus have ceased.
Behavioural signs of oestrus are due to the influence of oestrogens:
- restlessness
-drop in milk production
-standing to be mounted
-presence of clear mucus
-swelling and reddening of vulva
Metoestrus (Day 2-4)
10



oestrus ends
a corpus luteum (CL) is formed
progesterone levels in circulation begin to rise
Dioestrus (Day 5-17)



period of maximum corpus luteum size and function
high levels of progesterone in circulation
at the end of dioestrus, luteolysis of the corpus luteum begins.
11
Exercise-III: Detection of oestrus in farm animals.
Oestrus is defined as period of sexual receptivity, during which ovulation occurs in most
of the species and corpus luteum formation begins. Various methods used for oestrus detection
in farm animals are:
1. Physical symptoms
2. Prading of vasectomised bull and Androgenized heifers
3. Examination of external and internal genitalia
4. Examination of cervical mucus.
1. Physical Symptoms:
Animal in estrus exhibit some typical symptoms. On keen observation of these
symptoms estrus detection can be made. Some of the typical symptoms are : Bellowing,
Excessive movement, Frequent movement, Courtship behaviour, Mounting behaviour, Tail
raising, Frequent urination etc.
2. Prading of vasectomised bull and Androgenic heifers:
Bull prading method is used to detect animal in oestrus at organised farms. Vasectomised
bulls are taken to animal barn, where it approaches female in oestrus and gives Flehmen reaction
after sniffing vulva / licking urine of the animal in oestrus. On basis of bull prading animals so
detected in oestrus are separated and examined.
The culled heifers that have been treated with androgens can be used as oestrus checkers in
organised dairy farm rather than using altered bulls. The androgenized heifers are safer as these
female poses no disease threats. Continued treatment provides an effective animal for long
periods and androgen infections are easier and less expensive than some of the surgical
procedures.
3. Examination of external and internal genitalia:
The animals separated after bull prading showed following changes in external genitalia,
vulva is hypermic, congested and swollen. There may be some discharge from vulva. In internal
genitalia following changes may be observed.
Cervix – Open
Uterus – In tone
Ovary – Presence of graffian follicles.
12
5. Examination of cervical mucus :
Cervix is considered a glandular organ. Internal lining of cervix is of columnar
epithelium and near fornix of cervix is stratified squamous epithelium, which secretes mucus
secretions. These mucus secretions contain chloride ions, which forms fern pattern.
For examination of cervical mucus, collect the cervical mucus in a vial or directly take
on a glass slide, spread it. Dry in air. Examine under microscope. Fern pattern can be observed.
On the basis of degree of fern pattern formation stage of oestrus can be predicted.
Fig.1 Degree of fern pattern in cervical mucus of oestrus cow
(A)
Pr
(B)
Primary, secondary and tertiary branches in fern (A) 40X,
(B) 10 X magnification
13
Exercise IV : Semen and its components
Semen is composed of seminal plasma and spermatozoa. The quantity and sperm
concentration vary with species. Animals discharging large volume of semen have poor sperm
concentration. Total sperm discharged per ejaculate is 1000-1500 × 10
6
/ ml
in
bulls. For
artificial insemination two types of straws are used i.e. French medium straw of 0.5ml, which
contains 30 million sperms/ straw or 60 million sperms/ ml and French mini straw of 0.25ml
which contains 25 million straw or 100 millions of sperms/ ml.
Semen is composed of spermatozoa and seminal plasma. Its source is epididymis and vas
deferens, which provide most of the fluid portion (seminal plasma). In terms of the total volume,
the contribution of the epididymis and vas deferens is small. In bull the greatest contribution to
the fluid volume of the semen is from the vesicular gland, with minor contribution from the
prostate gland and Bulbourethral glands. In boars, there are greater contributions from the
prostate and Bulbourethral gland with smaller proportions from vesicular glands. These
differences are reflected in the chemical composition of the semen. Bull semen is higher in
fructose and sorbitol, which comes from the vesicular glands, whereas boar semen is higher in
most minerals, the major source of these being the prostate gland.
Spermatozoa
The concentration ( no. / ml ) of spermatozoa in an ejaculate of semen is approximately
150 millions for stallions, 200 millions for boars, 1.2 billions for bulls and 2 billions for rams.
Approximately 60 to 70 % of spermatozoa in the semen are expected to be progressively motile
with an average speed of 6 mm per minute. In high quality semen, 80 to 90 % of the
spermatozoa will have normal morphology, concentration; motility percent and morphology are
all important criteria in the evaluation of semen for artificial insemination. Spermatozoa of bulls
have and overall length of 60 to 70 µ.
The head is 8 to 10 µ long with tail accounting for the remainder. The head is flattened, about 4
µ wide and 0.5 µ thick. Both boar and ram have sperm of similar size, while sperm of stallions
are smaller ( about 50 µ in length).
The protein content is similar to the blood plasma in bulls. It contains high concentration of
electrolytes and does not contains glucose and cholesterol. Major substances present in semen,
which provides energy to spermatozoa are GPC, Fructose, Sorbitol, Inositol and Citric acid.
14
Exercise : Draw & label a diagram of sperm of cattle.
Signature of student
Signature of Instructor
15
Exercise V : Morphological Evaluation of semen
Objectives :
1. To use superior bulls to its full potential.
2. To reject inferior quality semen samples.
3. To evaluate the sexual status of bulls.
In morphological or physical or macroscopic examination of semen samples following
parameters are evaluated.
1. Colour
2. Transparency
3. Volume
4. Viscosity
5. pH
1. Colour :
Normal colour of semen sample is greyish white to yellowish white. In bulls and
rams
its creamish white. In some abnormal semen samples having red colour or brown
colour may be due to presence of blood and faecal contamination respectively. In some
samples high yellow colour may be due to excess of riboflavin.
2. Transparency :
A normal semen sample is opaque. However it may be translucent or cloudy due
to poor sperm concentration or due to infective semen.
3. Volume :
The volume of semen sample is generally constant with in species, however it
varies frequency of collection. The volume of ejaculate in different species of animals is as
follows.
Species
Average Volume ( ml)
Volume Range (ml )
Bull
4
1-15
Stallion
70
30-250
Ram
1
0.7-0.3
Boar
250
125-500
Dog
10
1-25
Cat
0.04
----
Cock
0.6
-----
16
4. Viscosity :
A thick creamy consistency for bull semen is considered to be excellent.
A thin creamy semen is considered very good and graded as +++.
A thick milky semen is considered as good and graded as ++.
A thin milky semen is considered as fair and graded as +.
A watery type of semen is of poor quality.
5. pH :
pH of freshly ejaculated semen ranges from 6.4 to 6.9 in buffalo – bull. Ejaculated semen
becomes acidic on storage due to metabolic activity of sperm. Semen sample with alkaline pH
is considered as of very poor quality.
Ex. Morphological evolution of Semen .
1. Colour ______________________________
2. Transparency ________________________
3. Volume _______________________________
4. Viscosity _______________________________
5. pH__________________________________-
Signature of student
Signature of Instructor
17
Exercise – VI : Microscopic evaluation of semen
Microscopic evaluation of semen is carried out by mainly three methods i.e.
1.
Progressive motility.
2.
Determination of sperm count.
3.
Determination of Live & Dead sperm count.
1. Progressive motility
The motility of a sample of semen is expressed as the percent of cells that
are motile under their own power. The progressively motile sperm is the one that is
moving or progressing from one point to another in more or less a straight line. Most of
the ejaculates show other type of motility. These include circular and reverse movements
due to a tail abnormality or vibrating or rocking type of movements often associated with
aging. Progressive motility is the most important individual quality test, because fertility
is highly correlated with the number of motile sperms inseminated. The percentage of
progressive motility can range from 0 to 80%. The usual motility determination is a
subjective measurement based on individual judgement. Semen samples with progressive
motility of less than 40% are not suitable for insemination except in case it is from a very
superior bull.
2. Determination of sperm concentration:
Sperm concentrations are expressed as number of cell per ml and must be
known on each ejaculate in order to make the maximum number of breeding units
containing a given number of motile sperms per unit. Sperm concentrations can be
determined mainly by two methods:
a) Direct cell count (Hemocytometer/Neubauer chamber method)
In this method Neubauer counting chamber is used for direct cell count.
This procedure is quite similar to TEC method. Thoma’s pipette used for TEC is used.
Procedure
Equipment required are:
o Neubauer counting chamber
o Thoma’s RBC pipette.
o Diluting fluid: i) Distilled water mixed with eosin.
ii) Chloramine-T
iii) Diluted formaldehyde solution.
o
Microscope etc.
18
Fix Neubauer containing chamber at low power of microscope to identify TEC area. In
Thoma Pipette suck semen sample upto 0.5 mark and then diluting fluid upto 101 mark.
Mix properly, discard a few drops and then charge the chamber. Allow few minutes for
setting thereafter count the sperms in five squares. Express your results as :
No. sperms / ml = no. sperm in 0.1 mm3 × 10 × dilution rate ×1000
Where 0.1
10
1000
area / volume of counting
to make value per micro litre ( µl)
to make valur per mililiter
This is time consuming and tedious method of determining sperm concentration.
The inexperienced persons should make two dilutions of a sample and count two
chambers from each dilution as a check on his technique.
Results:
a) Progressive Motility :-
b) Sperm Concentraion :
Date
Signature of Student
Signature of Invigilator
19
Exercise – VII : Determination of live and dead sperm count
Eosin is referred to as a differential stain in that it cannot pass through living cell
membranes but can pass through nonliving cell membranes. A background stain such as
nigrosin, opal blue, or fast green helps make the unstained sperm heads visible. The percentage
of live sperm in a sample of semen has been used as verification of motility determinations.
However it should be kept in mind that percentage of live sperms will always be somewhat
higher than the percentage of motility.
Staining Technique
Several staining mixtures have given good results. All of them contain about 1% eosin plus
one of the background stains. Preferred background stains are 2% green, 4% aniline blue or 5%
nigrosin. Both eosin and background stain are dissolved in 2.9% sodium citrate dehydrate
buffer. For slide preparation take one drop of stain and another drop of semen sample in a Petri
dish. Mix them with a glass rod. Take a little quantity of this mixture on a clean glass slide.
Spread it uniformly and dry it in the air. Examine it under oil immersion lens.
20
Exercise: Live and Dead Sperm Count.
Results:
Field no.
Live sperms
Dead sperms
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
Total =
Percent of live sperms
=
Percent of dead sperms =
Date
Signature of student
Signature of Instructor
21
Exercise: Complete evaluation of Semen
(A)
(B)
Morphological
1. Colour
------------------------------------
2. Transparency
------------------------------------
3. Volume
------------------------------------
4. Viscosity
------------------------------------
5. ph
------------------------------------
Mass motility
(C) Sperm concentration
------------------------------------
------------------------------------
(D Live –dead sperm count
(C)
Live
------------------------------------%
Dead
------------------------------------%
Remarks
------------------------------------%
Abnormalities:
------------------------------------
Date :
Signature of student
Signature of Instructor
22
Exercise VIII : Abnormalities of spermatozoa
Material Required:
1.Eosin – Nigrosin Stain
2.Microslides
3.Sample Pipette
4.Microscope
5.Semen sample
6.Lens Paper
Prepare the smear of the semen with eosin nigrosin as for counting for the livedead spermatozoa examine and observe individual sperm for morphological abnormalities and
classify them as follows.
1. Primary Abnormalities:
a. Head abnormalities:
Giant head (Macrocephalic)
Small head (Microcephalic)
Double head
Pyriform heads
Long and narrow heads
Loose abnormal heads
Asymmetrical heads
Primordial cells
b. Middle piece abnormalities
Double middle piece
Coiled middle piece
Swollen middle piece
23
Abaxial attached middle piece
Short or enlarged middle piece
Filiform middle piece
Vestigial middle piece
(c) Tail abnormalities
Tightly coiled tails
Double tails
Absent or short tails
Broken tails
Kinky tails
Truncated tails
II. Secondary abnormalities:
Loose or free normal heads
Detached galea capita
Proximal droplets
Distal droplets
Loose or free middle piece
Loose or free tail
Bent middle piece
Kinked middle piece
Coiled tails
Bent tails
Tufted tails.
24
Exercise : Examine and record different abnormalities of spermatozoa
1. Primary Abnormalities:
a. Head abnormalities:
b. Middle piece abnormalities
c. Tail abnormalities
II. Secondary abnormalities:
Date
Signature of Student
Signature of Instructor
25
Exercise IX: Measurement of hormone concentration in body fluids
Most of the hormones are present in the circulating body fluids and the tissues in
extremely minute quantity. Some hormones are present in as low quantity as one-millionth of a
milligram (pg/ml). Common methods used for estimation of hormones are:
1. Bioassay
2. Radioimmunoassay (RIA)
3. Enzyme Immunoassay (ELISA)
4. Enzyme multiple assay (EMIT)
1. Bioassay:
Bioassay means use of an appropriate animal preparation in which one can test
the action of the hormone. For instance an appropriate bioassay for ADH is to measure the
degree of water conservation caused by injecting plasma or an aqueous extract of the body tissue
from an experimental animal into a test animal that does not itself secrete ADH. And to
compare the animal’s response to known quantity of pure ADH
2. Radioimmunoassay :
RIA is based on the principle of competitive binding between radiolabelled
antigen present in the body fluid to be assayed for binding to limited amount of specific
antibody. Three main components required for RIA are fluid to be assayed, antibody, purified
hormone labelled with radioactive isotope.
The hormone present in plasma/serum competes for binding sites on the
antibody. The quantity of each of two hormones, the natural and radioactive will bind in
proportion to their concentration. Precipitating agent Polyethyel glycol (PEG) is used , the
antibody – hormone complex is separated from the reminder of solution and quantity of
Radiolabelled hormone that has bound with the antibody is measured by Gamma/Beta
counter. If large amount of radioactive hormone has bound it means that natural hormone
present is in very small quantity and vice-versa. With help of standard curve the concentration
of hormone in a unknown sample is measured. Incubation period at different intervals vary for
various hormones.
Drawbacks:
1. Radioactive materials used in the process is a health hazard.
2. Storing and disposal of radioactive material is also very difficult.
26
3. Enzyme Linked immunoassay (ELISA)
ELISA is an EIA used for antigen measurements. It ia a heterogeneous, solid phase assay
that requires the separation of reagents. ELISA has two available techniques for antigen
measurement, the sandwich technique and the competitive technique. The sandwich or double
antibody technique begins with an antibody bound to a antibody bound to a polystyrene well
plus the antigen to be measured. An enzyme conjugate is then added to the well with bound
antigen- antibody or immune complex. Next a substrate is added to the enzyme conjugate which
is bound to the immune complex. From here if there are changes due to the presence of the
enzyme conjugate bound to the immune complex, a positive test or colour change will occur.
The antigen competitive inhibition assay begins with an antibody bond to a polystyrene well
plus a test sample containing an antigen mixture to which an antigen-enzyme conjugate is added.
At this point competitive inhibition occurs between the antigen-enzyme conjugate and a
unlabelled antigen, depending on which antigen type is in excess, two different outcomes can
follow when binding to specific antibody occurs. After the formation of an immune complex
from an antigen-antibody binding, the reagents are separated by washing. Next a substrate is
added to the complex. If the antigen enzyme conjugate is the antigen in excess a colour change
will occur indicating that the substrate was chemically changed as a result of the enzyme
conjugate being bound to the immune complex. If it is the unlabelled antigen that is in excess
there will be a little to no change in colour because the test sample contains antigen-typespecific antigen.
4. EMIT ( Enzyme multiple assay)
Emit is an EIA used for rapidly assaying micro amounts of drugs and substances in
human biological fluids. It is homogenous, liquid phase assay that measures haptens for drugs,
hormone and metabolite determinations.
EMIT vs. ELISA
EMIT and ELISA have many characteristics that separate them from each other. EMIT is
mainly used in drug, hormone, and metabolite determinations. It measures haptens which are
small molecule while ELISA measures macromolecules such as antigens and antibodies and is
used for diagnosing infectious disease and immunoglobins. EMIT is faster than ELISA but
ELISA has greater sensitivity. Both of these EIAs have a long shelf life and can be done by
personnel with only minimal training.
27
Exercise:
Measurement of hormone concentration in body fluids
1.
Bioassay:
2.
Radioimmunoassay :
3.
Enzyme Linked immunoassay (ELISA)
4.
EMIT ( Enzyme multiple assay)
Date
Signature of Student
Signature of Instructor
28
Exercise - X : Effect of insulin on carbohydrate metabolism.
Immediately after a high carbohydrate meal, the glucose that is absorbed into blood
causes rapid secretion of insulin, which in turn causes rapid uptake, storage and use of glucose
by almost all tissues of the body especially by liver, adipose tissue and muscles. Insulin
increases the activity of glucokinase enzyme from liver.
It increases the activity of
phosphofructokinase enzyme that promotes glycogen synthesis. Rain cell normally uses only
glucose for energy. Therefore it is essential that the blood glucose level, be maintained above a
cricitcal level, which is one of the important functions of blood glucose control systems. Insulin
has either very little or no effect on uptake or use of glucose by brain cells. When blood glucose
level falls to 20 to 50 mg/dl (In Monogastrics) symptoms of hypoglycaemic shock develops.
Experiment
Select four calves randomly Record their body weight. Calculate dose of insulin @ 0.25
I.U./Kg. body weight. Collect control blood sample and inject the calculated dose of
insulin intravenously. Collect subsequent blood samples at interval of 5, 10 and 15
minutes. Determine the blood glucose level in all these blood samples and draw
conclusion from results.
Exercise :
a) Basic Blood glucose level :
b) Blood glucose level at
5 minutes –
10 minutes –
15 minutes –
Conclusion –
Signature of student
Signature of Instructor
29
Exercise-XI. Growth measurement in different species of domestic
animals.
The most common method of measuring growth in different domestic animals is
by way of recording height, length, girth and bone measurement
Following measures and measurements can be used to monitor growth in various
species of domestic animals. The term measurement is generally applied for height,
length, girth and bone measurement of animals.
1. HEIGHT:
Height of an animal is the vertical distance from the ground to the height point of
withers. It is measured in hands (a hand is equal to four inches) in case of horses and
in inches in other animals.
The measuring equipment consists of on upright stick
graded up to 18-19 hands or 72-76 inches and a cross rod carrying a spirit level. The
cross rod slides up upright down the upright stick. The following precautions must be
observed while recording the height of an animal.
1.
The animal should be made to stand on hard leveled ground.
2.
The animal should be made to stand on all its four legs i.e., with
equal weight
on each limb, i.e. the animal should stand square.
3.
Animals are measured from near side particularly horse.
4.
Keep the stick hidden when you approach the animal.
5.
If the animal is afraid of the stick, the man holding him should place his hand
over the eye.
6.
Let the cross rod slide slowly to the highest point of withers With a nervous horse
which is frightened by the touch of Cross rod, place your
hand on wither and
bring the rod gently down on it.
7.
Mention whether the horse is being measured bare foot or shoed.
8.
In case of cattle particularly Indian Cattle (Zebu) through the hump is at a higher
level yet for the sake of uniformity, the height is measured just behind the hump.
(withers) as the foreign cattle don’t
9.
have the hump.
When no standard stick is available, measure the height of animal by comparing
it with your chin or nose tip.
10.
When a large number of animals are to be measured it is better and convenient
to use wall graphs.
30
11.
Ordinary measuring tape can also be used to measure height by holding
it
tightly from the highest point of the withers to the coronet in a slanting position.
It will give the approximate height of horse.
B. LENGTH :
Length of an animal is measured in inches from the point of shoulder to the
point of buttocks in case of horse and in case of cattle, sheep and goat from the
point of shoulder to pin bone. Ordinary measuring tape is used for this purpose. If
measuring tape is not available the length can be measured by means of any cord
or rope and then it is measured by foot rule Wall graphs are also helpful in
recording length.
C. GIRTH:
Girth or circumference of the body of an animal is measured at girth place
which is just behind point of elbow with a measuring tape. The tape should neither
be held loose nor tight but with sufficient pressure and reading is recorded in
inches.
D. Bone Measurement:
To be sure that the animal is growing according to age and nutritional level,
the cannon bone of a horse is measured by
metallic tape. The off fore-leg is
held up and the measuring tape preferably metallic one is placed round the near
fore Cannon bone about 2” below knee. The tape should be drawn tight without
undue force. If there is some abnormality in the near fore Cannon e.g., splint etc.
Which can give false measurement the other fore-leg should be measured. If both
the legs are affected, no measurement should be taken and the fact be recorded in
the report.
Precaution
1.
In taking heart girth measurements, it is important that the animal is kept off
water for a minimum of 12 hours.
2.
The animal should be made to stand on all its four legs squarely under the body
and use the head up in normal position and the tape be passed
round
the
body just at the back of shoulder at the smallest circumference and pulled up
snugly.
31
E.
WEIGHT: The estimation of weight of an animal is useful:
(i) To determine its growth.
(ii) To determine the approximate weight of dressed meat.
(iii) To estimate quantity of fodder/ration required.
(iv) To calculate dose of medicine to be administered.
Procedure
1.
Use of Weigh Bridge: It is used to get the most accurate weight of the animals.
Before taking weight, make sure the needle is at O or the
Walk the animal over it and record the weight
2.
weigh
bridge
is
tared.
indicated on the dial.
Formulae for approximate Calculation of Weight: These are used when weighing
bridge is not available as is the case in field. The formulae used are:
(i)
Shaeffer’s Formula:
(A)
Live weight in Kgs. = L X G2 /660
Whereas L = Length in inches and
G = Girth in inches
(B)
Live weight in Kgs = HG2 x L/11200
Whereas L = Length in centimeters and
HG = Heart girth in cms.
(ii) Aggarwal’s Formula: Live weight in Kgs. = L x G/A
Value of A is 64.5 if the girth is less than 164 cm.
A is 61.0 , if the girth is between 165-200 cm.
A is 57.5, if the girth is over 200 cm.
The weight of young calves, sheep and goats is calculated by the Shaeffer’s
formula only. Cattle generally require fodder 1/10 of body weight and weight of dressed
Carcass, blood, hide, and fat is 2/5th,
1/13th, 1/8th and 1/12th of body weight.
32
Observations:
i.
Breed-
ii. Tag No
iii. Age
iv. Sex
v. BW-
Table; 1 Recording of body growth measurements parameters
Dates
of Age of calf Height
recordings
Length
Girth
Body
Ration
at
(inches/
(inches/
(inches/
weight
required
recording
cms)
cms)
cms)
of calf
(kgs)
(weeks)
Remark
(kgs)
33
Table: 2. Recording of physiological parameters
Date of
Res. Rate
recordings
(res./min.)
Rectal Temp.
(0C)
Heart Rate
Pulse rate
(beats /min.)
(pulse/min.)
Table:3. Recording of Environmental elements
Dates
of
recordings
Environmental Temp.(0C)
Inside shed
outside shed
Humidity (%)
Inside shed
outside shed
34
Formula for growth curves:
1. Actual weight gain= L X G2 /660 (inches) or HG2 x L/11200(cms)
Whereas L = Length
G = Girth
2 . Absolute weight gain=
W2-W1
________
t2- t1
Where W 2= Final weight gain
W 1= Initial weight gain
t1=Initial time (age) in weeks
t2=Final time (age)
3. Relative weight gain=
Where
W2-W1
_________
W1
W1= Initial body weight
W2= Final body weight
35
Exercise XII: Instruments and Equipment’s used for measurement of
physical environment / Climate.
The animal is intimately and inseparably associated with the physical environment
or climate from its conception till death. The independent environmental or climatic
factors which should be measured include:
1. Air temperature.
2. Atmospheric Moisture Content.
3. Air motion/wind speed.
4. Surface temperature of solid surroundings.
5. Metabolic Heat production
6. Surface temperature of the animal.
7. Rate of Water loss.
The instruments and techniques used to measure these are:
(1) Air temperature and Thermometry
The temperature of the air in our environment (ambient temperature) is the
single characteristic used to describe thermal environment. Temperature is defined as
a relative measure of the heat content (Enthalpy) of a physical system. Three types of
thermometers particularly useful in climatology are:
36
(a) Liquid-in-Glass thermometers:
Mercury filled thermometers are the most
common instrument for measuring air temperature or dry bulb temperature. The
precautions required even with this instrument are:
i) Shield the bulb of thermometer with a reflective material in direct sun-light.
Otherwise the result will be erratic because of heating by infra-red radiation. Heavy
aluminum foil fastened loosely with cellophane tape works well.
ii) Shielding should not restrict the flow of air around the bulb.
iii) Take observations at several spots around the animal and the mean of there.
37
(b) Thermoelectric Thermometer: This class of instruments utilize the phenomena that
on electric current flows continuously in a closed circuit of two dissimilar metals
(thermojunctioins) when the junctions of the metals are maintained at different
temperatures.
The current generated is a function of the temperature difference
between junctions and it can be measured with the help of galvanometer or
potentiometer. For measurement in biological and meteorological range, copper and
constantin (Copper-Nickel alloy) thermo junctions or thermocouples are recommended.
(c) Thermistors: The thermistor thermometer is a highly useful device. The principle on
which these function is that certain semi-conductors exhibit a pronounced change in
electrical resistance in response to
slight change in temperature. Measurement of
this change in resistance provides a measurement of temperature at the thermistor.
Battery-powered thermistor thermometer makes a rugged
portable instrument well
suited to bioclimatology. Thermistor beads can be placed in hypodermic
needles
take sub-dermal temperature and in
temperature
steel rods
for
ground
to
measurement at various depths. The instrument should be periodically recalibrated as
the semi-conductor material may undergo slight change with aging.
2. ATMOSPHERIC MOISTURE CONTENT
The amount of water vapours in the air i.e., humidity controls the rate of
evaporation of water from skin
surface and from other moist tissues e.g., lungs,
respiratory passages and conjunctiva of eyes etc. The rate of air motion and degree of
saturation of the ambient air governs the rate of evaporation.
If the air is already
saturated, no evaporation can occur. Relative Humidity is defined as the amount of
moisture in the air as compared with the amount that air can contain at that
temperature.
It is usually expressed in percentage.
The psychometric chart is a
convenient graphical representation of the mathematical interrelationships of
temperature, volume, total heat content, dew point, absolute humidity, relative
humidity etc. Absolute humidity is defined as amount of water vapours in air at that
temperature.
38
Atmospheric moisture content can be measured by:
a)
Sling Psychrometer consists of two thermometers clamped in a frame
turn is fastened to a swivel handle. A wet silk wick covers
which in
the bulb of one while the
other thermometer is bare hence called wet bulb and dry bulb thermometers. When the
instrument is rapidly whirled water evaporates from the wick, cooling the bulb. The rate
of evaporation from the wick is a function of the vapor pressure gradient
determining
in turn the depression of the wet bulb thermometer reading below the dry bulb. From
the psychrometric metric chart or table, the vapor pressure can be read directly. The
precautions include.
39
1.
One minute swinging adequately cools the wet bulb to its lowest reading. If
temperature is continuously falling, then repeat swinging a few times.
2.
There should be no obstructions in the path of the swinging thermometers.
3.
Distilled water should be used as it prolongs the life of wick. If
wick is dirty, it
should be washed with detergent and rinsed thoroughly with distilled water.
40
b) Motor Drives Psychrometer: These battery powered aspiration
psychrometers are
of several types available for field and conventional use. These provide the same
results as the sling psychrometer and the air motion is caused mechanically instead of
whirling across the thermometer bulbs.
c) Dew point Hygrometer: These instruments indicate the dew point temperature by
providing a means for cooling a polished surface. The temperature at which moisture
just begins to condense on the surface (detected by the close appearance) is the dew
point. The surface of
dew point hygrometer cools the air sufficiently so that
moisture in it reaches saturation line and condeness.
d) Hair Hygrometer: Human hair absorbs and desorbs moisture with change in
atmospheric humidity.
The length of hair under tension changes in turn with its
moisture content. This motion is converted through a system of levers to a pointer
indicating the relative humidity.
Fitted with a pen, the pointer records the relative
humidity on a revolving drum.
41
3. AIR MOTION/WIND SPEED AND ANEMOMETRY
The movement of air or wind causes the heat transfer by convection and
evaporation between the animal and environment. The units for the wind speed is
distance per unit of time e.g., Kms./hour or Kms./minute etc. are suggestive of mass
movement past a point. Turbulent air with little net mass movement has
power. Instruments which depend upon rate of cooling and are
high cooling
non-directional
like
thermo anemometer can measure this. Directional and (propeller, swinging vane)
semi-directional (cup) anemometers can be used to measure the wind velocity provided
air moves consistently from a known direction.
1.
Propeller and Cup Anemometers:- These instruments convert the kinetic energy of
the moving air mass into rotary motion of an impeller. The rotating impeller or propeller
or cup is connected to a D.C. generator and thus the current generated drives a
recording per or on indicating meter calibrated directly to read the wind speed, directly
in
meters/second or Kms./hr. It is very well suited to meteorological
observations
but the disadvantage of these for microclimate of the animal is that they are directional
end don’t indicate air speed accurately at low wind flow.
42
2. Swinging Vane Anemometers: The velocity pressure of the moving air stream is
converted to static pressure against a counter weighed vane in the velometer.
It is
popularly used in the heating and ventilating field for the measurement and adjustment
of air distribution systems.
3.
The thermo anemometers used for measuring wind velocity include:
(a) Wilson Thermoanemometer
(b) Alnor thermoanemometer
(c) Anemotherm
(d) Heated globe Anemometer.
4.
Kata Thermometer: It is used to determine cooling power of air as a
measure
of
efficiency of ventilation in factories, mines etc.
43
(iv) Atmospheric Pressure: The weight of atmosphere at the earth’s surface exerts a
considerable squeezing force.
The atmospheric pressure varies with change in
altitude, temperature of air and the meteorological
phenomena.
The
standard
atmospheric pressure unit agreed internationally is 760 m.m. of Mercury Column
corresponding to the mean atmospheric pressure at sea level. Another common unit of
atmospheric pressure is millibar.
One millibar is defined as the pressure of 1000
dynes/cm2. For conversion 1.013 millibar = 760 m.m. Hg.
The instrument used to measure atmospheric pressure is called BAROMETER
and these are of following types:
(i) Mercury Barometers: The basic principle in all barometers is the balancing of the
atmospheric forces against some known resistance. In
the pressure of the air balances a column of
mercury.The
the mercury barometer,
simplest
mercury
barometer was discovered by Torricelli. It consists of a sealed glass tube completely
filled with mercury inverted into a reservoir containing additional mercury. The pressure
of the air on the surface of the dish supports the column in
the tube.
Space above
mercury in the tube is a complete vacuum. Thus the only downward force on the
column comes from the weight of mercury. A correction for temperature is also applied.
If temperature fluctuation is regular and varied, then the correction should be applied
making use of correction table supplied along with. In modern barometers, the height
of column above reservoir is either already marked by a scale or in measurement by
Vernier scale.
(ii) Aneroid Barometer: This instrument doesn’t contain any liquid. The heart of this
instrument consists of an evacuated pressure sensitive capsule. The top is supported
by a stout spring to prevent collapsing. Variation in external. pressure causes the top to
move slightly up and down.
A system of levers and gears magnifies this
movement moving a pointer over a scale. A correction must be made for temperature,
gravity and relative humidity.
(iii) Microbarograph: It is a portable recording aneroid type barometer. The motion of
the pen is damped by two oil-filled dash pots which
eliminate motion due to shock
44
and vibration. The spring drives drum revolves under the pen to provide a continuous
record of atmospheric pressure.
(iv)
Measurement of Light: The animal eye is sensitive to a narrow based of
wavelengths in the electromagnetic spectrum. Light causes heating
characteristics of wavelength duration and intensity
are
also
effect but the
important.
It
is
measured with the help of Light meters. The Common direct reading light meters use a
selenium photovoltaic cell. Illumination striking the cell causes a current to flow, a
meter in the circuit indicates the light intensity of a source measured directly in meter
candles.
Spectrophotometers: These work on principle that light energy at a selected
bond is measured by use of appropriate filters over photoelectric cells.
wave
For more
precise analysis of the spectral quality of reflected or transmitted light, a recording
spectrophotometer can be used.
Sun-shine Recorders: Campbell - stokes apparatus adopted by World Meteorological
Organization is used for measuring sun-shine. A water filled glove concentrates the
solarays and burns traces on a special card. Length of burn is on index of length of
sun-shine. Great care should be taken in placing the instrument and selection of cards
to permit proper interpretation.
(v)
Measurement of Noise: Noise is an environmental stress of increasing impact in
our technologically advanced society. Its effect is well demonstrated by the reports of
45
losses of milk production in dairies situated near jet airports. It is measured by sound
and acoustical instruments which include:
1. Sound Level Meters: The unit used to measure sound is decibel (db). The fighter
jets while taking off produce a noise of 120 db. Sound
sound level which is a measure of apparent
level
meter
measures
loudness which varies with frequency.
Sound pressure level is a physical measurement of the root mean square variation in
pressure of
the waves measured in microbars. Sound level measurements are made
with the frequency response weighted to match the response of the ear.
2.
Microphones: Ceramic-type microphones are usually standard with sound level
meter but if temperature of surroundings is high, condenser type microphones should
be used. It should be recalibrated periodically.
46
Exercise-XIII. Basic Requirement of a meteorological station/
Observatory.
The most important climatic variables are temperature, humidity, wind, sunshine, rain-fall and evaporation. For the field meteorological station/observatory, the
essential requirements for making useful meteorological observations are:
(i)
The site should be representative of the Animal-soil-crop climate
conditions for which data is intended.
(ii)
Each instrument should give a reliable measure of meteorological variable; they
should be easy to maintain.
(iii)
Observations should be easy to make and to record.
(iv)
The observers should be reliable and properly trained.
(i) Selection of Site: It should be representative of the intended conditions and as large
an area as possible.
It shouldn’t be an area of abrupt climatic differences e.g.,
swamps, mountains, river, gorges and lakes etc. to measure local weather phenomena
such as humidity, wind and precipitation etc., additional stations should be installed.
The site should be on level ground, away from any road, free from obstructions like
houses and big trees etc. It should be easily
approachable by observers to ensure
punctuality and reliability of observations.
The station should have slow growing
drought resistant grass cover. It should be 10 meter X 10 meter in dimensions and be
centrally located in open area of 50 X 50 meter. It should have 1.2 meter
high
wire
mesh fence fixed to posts 2 meter high and 2.5 metros from each other with a hinged
door at the center of East-West fence with a narrow path North to South to keep
observer away from self- recording instruments. Some of factors which have effect on
weather station include:
(i)
Vegetation : The vegetation is more in irrigated areas compared to dry areas
resulting in lower temperature, higher humidity and decreased evaporation.
(ii) Topography: Elevation differences affect precipitation, minimum temperature, wind
speed and wind direction.
47
(iii) Rivers:
These have less effect confined to 100 metros except for large rivers and
deltas.
(iv) Lakes:
(v) Sea:
Depends on size of lake but generally effects 1 to 2 Kms.
Effects are very rapid in first 2 Kms but are there upto 10-15 Kms. Affect
wind speed, humidity and temperature.
(vi) Altitude:
With increase in altitude temperature and evaporation decrease while
rain fall and wind tend to increase.
(vii) Mountains: Down wind effects up to distances 50 times their height.
(ii) INSTRUMENTS REQUIRED:
(a) Stevenson thermometer shelter - Used to place instruments for measuring air
temperature and humidity.
(b) Maximum-Minimum air temperature - Consisting of Mercury-in-glass maximum
thermometer set for daily
thermometer.
OBSERVATIONS.
(c) Humidity to be recorded
- One wet bulb thermometer with wick and (twice daily)
small water reservoir. One dry bulb thermometer both are mercury-in-glass.
(d) Rain fall (Daily)
- Rain Gauge Receiving area is 200-500
Cm. Capacity upto 200 mm. of rain with
graduated measuring cylinders.
or
Self recording clock rivers rain gauge apparatus.
(e) Wind flow or Speed
-
Cup counter Wind Anaemometer 2 meter
and direction (Daily)high with a speed range of 10,000 Kms. with a swinging or
Wind Vane.
48
(f) Sunshine (Daily)
- Campbell stokes Sun-shine recorder with
leveling devices and recording Cards.
(g) Evaporation (Daily)
- U.S. Open Pan Evaporimeter Non-Corrosive
with still well hook gauge and metric or
caliper.
Weather Conditions
-
Visual Observation on cloud cover.
Visibility, Hail occurrence, Wind direction etc.
Along with these, direct reading instruments should be installed to check accuracy
and to calibrate these instruments.
Observations should be taken regularly and punctually. Readings must be made each
day at the same time. Exact time of actual observation must always be noted on the
daily record sheets. Observer should follow the same set routine while taking
observations. For this purpose daily routine sheet should be drawn for specifically each
station for type and order of observations, time markings, change of cards, filling of
reservoirs and checks to be made etc.
Each station should maintain observation sheets for daily, monthly and yearly data A
complete set of records should be kept at the station.
49
Exercise XIV Climatic Change related to environmental physiology
and the effect and
relationship of environment / weather on animals
The effect of weather or environment can be in form of
(i) Effect on livestock health and production.
(ii) Animal Diseases and Parasites.
(i)
Effect on Livestock health and production
It includes environmental
problems of livestock housing, health and
production. Apart from its direct effects weather affects farm animals through
the crops on which they feed and the ground on which they are kept. It affects
their
feeding, growth, fecundity and health, their geographical distribution, the
yield, quantity and quality of animal products,
the preparation of these products
and their capacity for storage and transport. Genotype environment interactions
in cattle are generally assumed to exist.
The performance of beef cattle in
relation to their environment needs to be further investigated world wide. In
temperate zones, livestock is less sensitive than plants to climatic stress. Some
processes however such as milk production are fairly closely related to
temperature and moisture.
(ii)
Animal Diseases and Parasites
Direct and indirect effects of weather on the various types of animal
diseases, injury and death, economic losses, forecasting incidence and intensity
of animal diseases are included in it. Meteorological factors can influence animal
diseases in various ways by affecting.
(a)
The resistance of hosts to germs/pathogens
(b)
The resistance and evolution of these germs/pathogens during their
biological
cycle particularly when part of this cycle occurs outside the host animal.
(c)
The conditions under which control measures are applied.
Knowledge of the interrelations between animals, weather and certain diseases
is adequate to provide animal disease forecasts which are available in some countries.
These services should be extended to include other animal diseases and more
countries.
50
Exercise XV To record various physiological parameters and study
the effect of extreme weather on animal.
Before Exposure
(I)
Temperature
(ii)
Respiratory Rate
(iii)
Heat Beat
(iv)
Pulse
(v)
Salivation
(vi)
Water Intake
(vii)
Any Other Response
2.
After Exposure for 1hour.
EFEECT OF COLD:- Behavioural adjustments to the cold environments tends to
be just the reverse of those made to hot climate. Efforts are made to increase internal
heat production and decrease the steep thermal
gradient between the body’s core
and the environment. Body surface is minimized to conserve heat, rate respiration is
reduced to decrease the heat loss from the surface of respiratory tract, motor activities
are increased. However if these efforts fail to maintain body temperature, they tend to
cease. The consequence is death for all homiotherms except those, few which escape
the riggers of cold environment by hibernating.
Most domesticated animals are
physiologically and behaviourally better equipped too deal with cold environments than
the hot ones since in general they have evolved from temperate zone stock.
51
The behavioural adjustments of mammals to cold environment is as follows:-
(i)
To keep body temperature normal, (a) there is increased heat production
through increased food consumption and motor activities. (b) With the help of
thermal radiation by seeking direct sun-light and warmer bodies i.e., group
huddling.
(c)
The evaporations decreased through decrease in respiratory rate, decreased
body surface (flexure) and minimization of air movements over
body
surfaces.
(ii)
To counter moderate but not a critical decrease in body temperature
(a)
Heat production is increased through hyperplasia, shivering and decreased
general activity.
(b)
Thermal radiation as mentioned above.
(c)
Evaporation is decreased by depressing respiration and increasing body
flexure to maximum.
(iii)
To counter critical decrease, no warming strategy is adopted. There is cessation
of shivering, depressed metabolism, lethargy and reduced
mental
Under artificially controlled environmental
conditions,
temperatures
0oF greatly increased the heat production
and feed consumption in Zebu
cattle. While there is no appreciable
alertness.
near
increase in heat production in Holstein
Friesians while smaller European breads like Jersey exhibit moderate increases
in heat production and feed consumption under similar conditions. Common and
well-known
behavioural responses to cold temperature in all our domestic
animals
are increase in metabolic rate and food consumption, muscular
tone is
raised with apparent shivering and animals seek protective shelter
against wind and rain and huddle together to conserve warmth.
3.
EFFECT OF WATER DEPRIVATION:- Water and salts are lost from the body
continuously. The intake of water is governed by the thirst mechanism. The sensation
of thirst results from a dryness of the mucosa of the mouth and pharynx, which in turn is
due to a reduced flow of saliva in the thirsty animal. However the volume of fluid
52
ingested depends also
upon the amount of water absorbed from gastrointestinal
tract. The hypothalamus is known to be involved in regulation of water intake. If the
temperature-regulating center in the anterior hypothalamus is warmed during exercise
or hot environment, the animal drinks more water than normal.
Cooling of
hypothalamus has the opposite effect and the destruction of anterior hypothalamus
also results in reduced water intake.
If the water is withheld from animal, it results in excessive loss of water from the
body because water is continuously lost from the skin and respiratory tract through
evaportion and along with salts, It is lost in urine and faeces. The consequence is
finally dehydration of the animal. Under these conditions, animal will attempt to
conserve water by producing more concentrated urine and levels of ADH in the plasma
increases. This property is better in sheep in comparison to cattle.
Conservation of water is also achieved through reduction in food intake which
reduces the quantity of waste materials to be excreted in urine and hence the amount
of water needed to accompany these materials is less. The feces of Goat and Camel
are the finest examples of water conservation by preventing its loss in feces. Absence
of water in feces make them hard and pellet type. The metabolism rate of the animal
decreases which also helps to keep the mine vol. minimal. There is less water loss by
evaporation from skin and respiratory tract.
There is increase in concentration of
plasma and reduction in its volume leading to increased colloidal osmotic pressure as a
result of which fluid enters the plasma from the interstitial space. Water from GIT tract
is absorbed and rate of salivary secretion is reduced. Camels and sheep can tolerate
27-32% loss of their body weight during dehydration. Water consumption for cattle in
hot environment is governed by the severity of heat and amount of dry matter eaten.
Studies in climatic chambers on Ayreshire steers which were deprived of
drinking water for 4 days at 15oC showed a decrease in feed intake, excretion of urine
and feces, evaporation, body weight, heat production, respiratory ventilation, plasma
potassium and urinary output. At the some time, there is a corresponding increase in
haematocrit, plasma total solids, plasma Sodium and Chloride, blood urea and urinary
sodium output. Dehydration for 2 days at 40 oC also elicit similar changes in most of
these physiological variables. Reduced water consumption in Indian Steers to one half
53
the normal amount resulted in a slower passage of digesta and increased absorption of
water from terminal section of the GIT.
If the animals with normal intake of water are subjected to heat, there is
immediate sweating but in dehydrated steers (4 days), the sweating occurred after a
delay of 1.5 hours with a resultant increase in body temperature.
There is no permanent change or any impairment of health due to short period of
dehydration. The restoration to normal of the various physiological values deranged by
dehydration generally requires more than 1 day.
animals at 400 C.
Rehydration with cold water of
The rate of fall was maximum in skin and minimum in rectal
temperature. Respiratory rate decreased from 130 to 40/minute.
Rehydration with warm water resulted in no change in body temperature but
caused respiratory rate to rise from 130 to 180/minute within 30 minutes.
To study the effect of dehydration, note down the following parameters:
S.No
Parameter
1.
Respiratory Rate
2.
Heart Rate
3.
Body Temperatu re
4.
Salivary Secretion
5.
Muzzle Secretion
6.
Body Weight
7.
Water Intake
8.
Any Other
9.
) Faeces(Quantity
Before-dehydration
After -dehydration
and Consistency)
10.
Urine
(Quantity
and Consistency
54
4.
EFFECT OF FOOD DEPRIVATION:- Adult animals maintain body weight at the
same level for years sometimes, despite great variation in caloric expenditure. Thus
feed intake must be adapted to caloric expenditure in adults. Similarly young animals
grow at well-defined rates in spite
of variability of energy expenditure suggesting
that there must be an adjustment of feed intake to requirements during growth period.
Effects of feeding have been explained by several hypothesis like
(a)
The thermostatic hypothesis: The specific dynamic action of feed increasing the
heat stress of the body as a whole.
(b)
The Glucostatic hypothesis:- The availability and utilization of glucose from body
fluids.
(c)
The Lipostatic hypothesis:
The concentration of circulating metabolites is
influenced by the fat in the depots.
(d) The concentration of serum amino acids.
Regulation of feed intake appears to be correlated with regulation of water
balance. Animal given no water, eat little or no dry food while those given no food drink
little or no water.
Hunger, appetite and satiety which are associated with feed intake are functions
of the central nervous system. Regulation of food intake is through complex neuro
physiological systems evident in hypothalamus and other parts and also for satiety
center or inhibitory mechanisms for the feeding responses. The lateral hypothalamic
area may be designated as feeding center’ and ventromedial area as ‘Satiety
Centre’.
Electrical stimulation of lateral hypotholamus markedly increase feed intake
while that of ventromedial area decrease the feed intake in cats. If certain points at
posterior hypothalamus are ablated or destroyed selectively, it leads to significant but
less profound changes in feed intake.
Pathological or Experimental lesions of
Ventromedial hypothalamus with associated obesity is primarily due to hyperphagia or
overeating.
Deprivation of food for a long time will cause a distinct weight loss, weakness,
emaciation and depression of general activities of the animal while hunger for a short
time will significantly effect the rumino-recticular movements, glucose level and VFA
levels. To study the effect of hunger, following parameters should be noted.
55
Before Deprivation
After Deprivation
1. Rumino-reticular Movements
______________
_____________
2. Blood Glucose Levels
_______________
_____________
3. Respiration
________________
_____________
4. Water Intake
________________
_____________
5. Blood Temp.
_______________
_____________
6. Heart Rate
______________
_____________
_______________
_____________
7. Body weight
5. EFFECT OF EXERCISE:- The exercise to the animal leads to mental
stimulation
and there is release of Epinephrine and Nor epinephrine which leads to increase in
heart rate, pulse, blood Pressure, Respiration to meet increased demand of Oxygen by
tissues., Besides that it the sweating leading to loss of water through evaporation, Heat
production body temperature. So the following parameters should be observed:
Before Exercise
After Exercise
1. Heart rate.
_______________
______________
2. Pulse rate
_______________
_______________
3. Body Temp.
_______________
_______________
4. Respiratory Rate
_______________
______________
5. Sweating
_________________
______________
6. Salivation
________________
_______________
56
6. Effect of Fear and Excitement: The fear will cause CNS stimulation causing the
animal to be excited. So the effect of fear and excitement is going to be the same as of
the exercise. So the same parameters should be observed.
7. Effect of Bath:
The bath given to the animal in summer provides it relief by helping it to lose the
heat. So the body temperature would
be decreased. But a bath in winter is likely to
add more stress as animal has to maintain its body temperature.
So it will show
increase in heart rate and muscular shivering etc.
57
ANNEXURE – I
COMPARATIVE ANATOMY OF FEMALE REPRODUCTIVE TRACT OF
VARIOUS SPECIES
ORGAN
COWS
EWE
SOW
MARE
Ovary
Almond shaped
Almond shaped
Berry shaped
Kidney shape
Active Ovary
Right
Right
Right
Left
Number of
Graffian
Follicles
Length of
Uterine tube
(cm)
Uterus
1-2
1-4
10-25
1-2
25
15-19
14-30
20-30
Bipartite
Bipartite
Bi-cornuate
Bipartite
Length of
8-10
4-10
10-23
7-8
25-30
10-14
10-23
20-35
Cervix (cm)
Anterior
vagina
58
Annexure – II
DURATION OF ESTROUS CYCLE AND ITS PHASES IN DIFFERENT
SPECIES OF ANIMALS
Species
Cow
Ewe
Sow
Mare
Goat
Bitch
Oestrus
21 days
17 days
20 days
22 days
21 days
4-7 months
Estrus
12-18 hrs
24-36 hrs
48-72 hrs
4-8 days
30-40 hrs
7-9 days
Metestrus
3-5 days
2-3 days
2-3 days
2-3 days
2-3 days
80-90 days
Diestrus
10-14 days
10-12
11-13 days
10-12 days
13-15days
-------
cycle
days
Pro-estrus
3-4 days
2-3 days
3-4 days
2-3 days
2-3 days
9 days
Time of
10-12 hrs
after end of
estrous
24-36 hrs
after
begins of
estrous
35-45 hrs
begins of
estrous
1-2 days
before end
of estrous
cycle
30-36 hrs
before end
of estrous
cycle
During 1-3
days of
estrous
cycle
ovulation
59
Annexure – III
SEMINAL CHARACTERISTICS IN DIFFERENT SPECIES OF ANIMALS
Species
Volume of
Sperm count /ml
Spermatozoa
Site of semen
ejaculate (ml)
×10 6
size
deposition
H
M
T
9
13
44
Bull
5-8
1000-1500
Ram
0.7-2
1500-3000
Stallion
75-100
100-150
7
10
42
Body of uterus
Boar
200-300
100-150
8
11
30
Uterus
Dog
2-16
10-200
7
10
44
Vagina
Cat
0.01-0.2
100-5000
---------
Vagina
Fowl
0.4-1
800-6500
---------
--------
Seminal Plasma –
---------
Body of uterus
Uterus
Acidic in bulls and rams
Alkaline in boar + stallion
Principal ions
-
Na & ClCa, Mg – small quantities
K substantial amount in spermatozoa
Buffering agent –
Hco3- ( source – vesicle gland )
Energy source –
fructose, sorbitol ( sugar alcohol)
GPC ( glyceryl phosphoryl choline)
Fructose
–
Bull, Ram semen high conc.
Stallion, boar – low conc. ( i.e. why storage capacity is low as compared
to bull + ram semen)
60