This is an important topic of mammalian (Male) reproductive toxicology.By doing this test sperm abnormalities should be cured. This topic is available in net but not like, what a master student try to find out.If there is anything wrong then correct me please. In this ppt i have included methods of semen analysis and the importance and some agents which create semen abnormalities.

1. INDIRA GANDHI NATIONAL TRIBAL UNIVERSITY
AMARKANTAK, M.P.
PRESENTATION TOPIC:-SEMEN ANALYSIS
SUBMITTED BY:-
GUTHAL BASUMATARY
M.Sc. 3rd Sem
DEPARTMENT OF ZOOLOGY

2. semen
• Semen also known as seminal fluid is an
organic fluid that may contain spermatozoa.
• Semen is produced and originates from the
seminal vesicle, which is located in the
pelvis.
• It is secreted by the gonads and other sexual
organs of male or hermaphroditic animals and
can fertilize female ova.
• In humans, seminal fluid contains several
components besides spermatozoa, proteolytic
and other enzymes as well as fructose are
elements of seminal fluid which promote the
survival of spermatozoa and provide a
medium through which they can move or
swim.

3. Semple collection(Semen)
• Methods used for sperm collection- 1.
masturbation, 2. condom collection, 3.
epididymal extraction
• 1. Sexual abstinence for at least 2 to 5 days
before collection of the sample
• Masturbation is only the suitable and best
method for the semen collection.
• Collect the sample (semen specimen) in a clean
wide mouthed plastic or glass container.
• Label the specimen with names, date, and time
of collection.
• Hold the sample with carefully and maintain at
room temperature.
• The specimen must arrive at lab within one
hour.

4. Importance of semen analysis
1. To investigate male infertility
2. To verify the vasectomy successful.
3.Used in stud farming (horse breeding) and farm animal breeding.
4.To observe the abnormality of sperm.
5. To detect the low no of sperm and volumes.

5. NORMAL RANGE OF SEMEN

6. SEMEN ANALYSIS
1. Semen analysis is also known as “SEMINOGRAM”.
2. Semen analysis represents a critical components of the initial evaluation
of the infertile male and is indicative of normal and abnormal
spermatogenesis, sperm morphology and sperm motility.
3. In combination with specialized sperm function test, it can be good
measure of infertility.
4. Semen analysis can be divided into two types
1. Macroscopic
2. Microscopic

7. MACROSCOPIC TEST OF SEMEN ANALYSIS
Following observations are come under this test.
1. LIQUIFICATION TEST:-
• Keep the semen sample in incubator in order to liquefy.
• Incubator is maintain at 37◦C temperature semen is produced.
• Pick the semen in dropper if not so it means it is abnormal.
• If it is sticky means it is abnormal.

8. 2.VOLUME:-
The normal volume of ejaculate after 2 to 5 days of sexual abstinence is
1.5 to 5 ml. The volume of the semen is measured by measuring dropper or
measuring cylinder.
3. COLOUR:-
The normal colour of the semen is greyish white.
4. pH OF SEMEN:-
Normal semen pH is in the range of 7.2-8.2 and it tends to increase with
time after ejaculation. Changes are usually due to inflammation of the prostate
or seminal vesicles. pH of the semen is test by using the pH parameter.
5. VISCOSITY:-
Failure to liquefy is usually a sign that there is inadequate secretion by
the prostate of the proteolytic enzymes fibrinolysin, fibrinogenase, and
aminopeptidase.

9. MICROSCOPIC EXAMINATION
Following observations are the microscopic examination:-
1. TOTAL SPERM COUNT:-
Over 15 million sperm per milliliter is considered normal,
according to the WHO in 2010.Older definitions state 20 million.
A lower sperm count is considered OLIGOZOOSPERMIA. A
vasectomy is considered successful if the sample
is AZOOSPERMIA (zero sperm of any kind found.
The sperm can be count by using HEAMOCYTOMETER or
MAKLER CHAMBER. It is observe under the microscope.

10. 2. Sperm motility
• It is study of moving sperms in any ejaculated
sample
• Take sperm into the sperm counting chamber
• Usually 20x magnification is used for counting
sperm cells and Sperm motility classified into-
a) Rapid progressive motility (ie, >25 μm/s at 37°C
and >20 μm/s at 20°C; Note: 25 μm is
approximately equal to 5 head lengths or half a
tail length).
b) Slow or sluggish progressive motility
c) Nonprogressive motility (<5 μm/s)
d) Immotility
• Sperms show < 50% rapid progressive and slow
progressive to achieve fertilization

11. 3. MORPHOLOGY TEST
• It is deal with shape size and appearance of the sperm
• Characteristics should be assist by carefully observing a stained sperm sample under the microscope
Procedure:-
1. takeout semen and drop in a slide
2. it is smear and fixed with ethanol ether and keep for 5-10 minutes
3. Then the sample is dipped into the different level alcohol . Starting from 80%, 70%, to 50% till it
reaches the distilled water . This is called as Hydration.
4. Now it have to be stain in Haematoxylin stain after keeping it for 3 minutes in taggotor.
5. Now reverse the process. Dip the slide in 50%, 70%, 80% and 90% alcohol. This is called
Dehydration
6. For better staining, the sample should be dipped in Orange G for 3 minutes.
7. Then after dipping in 90% alcohol stain the sample in EA50
8. Again dip it in90% alcohol followed by Xylene
9. This sample is taken to microscope and just observe it at 100x magnification and Emergent oil
is applied for perfect assessment
10. 10. Several shapes, size and forms of sperm will be observed
11. They will seen in normal forms, abnormal head, abnormal neck and abnormal tail.

12. • Head defects: Large, small, tapered, pyriform,
round, amorphous, vacuolated (>20% of the
head area occupied by unstained vacuolar
areas) heads with small acrosomal area (<40%
of head area), double heads, any combination
of these.
• Neck and midpiece defects: Bent neck;
asymmetrical insertion of midpiece into head;
thick, irregular midpiece; abnormally thin
midpiece; any combination of these.
• Tail defects: Short, multiple, hairpin, broken,
bent, kinked, coiled tails, or any combination
of these.
• Cytoplasmic droplets: Greater than one-third of
the area of a normal sperm head.

13. Disabilities related with semen
• Problems with sexual function — for example, difficulty with ejaculation or
small volumes of fluid ejaculated, reduced sexual desire, or difficulty
maintaining an erection (erectile dysfunction)
• Pain, swelling or a lump in the testicle area
• Recurrent respiratory infections
• Inability to smell
• Abnormal breast growth (gynecomastia)
• Decreased facial or body hair or other signs of a chromosomal or hormonal
abnormality
• A lower than normal sperm count (fewer than 15 million sperm per
milliliter of semen or a total sperm count of less than 39 million per
ejaculate)

14. FACTORS AFFECTING SEMEN QUALITY
• Bisphenol A (BPA)
BPA is used in industries to synthesize polycarbonate and epoxy resins. Since the
1960s, it has been used in the manufacturing of plastic bottles, suction cup, inner coating of
food and beverage cans, and so on.
• DDT
As an effective pesticide, DDT was widely used in agriculture and forestry.
• Dioxins and dioxin-like compounds:-
Dioxins had been shown to exhibit antiestrogenic activity.
• Heavy metal:-
It is beyond argument that cadmium and lead can induce male reproductive toxicity.
World Health Organization (WHO) indicated that even low-level exposure to lead and
cadmium (400 μg/l and 10 μg/l, respectively) can enable the semen has a significant quality
descend, although it did not show conclusive evidence of male hormonal changes in
reproduction.