1. The document discusses a dissertation on the impact of Clinostomum complanatum infection in the freshwater fish Nandus nandus, with a focus on histology. 2. Methods used to collect parasites from infected fish organs and prepare histological slides are described. 3. Results showed a prevalence of 47.36% infection in examined fish, with the highest intensities observed in November and December. 4. Histopathological analysis revealed destructive effects on the liver, ovaries, and spleen tissues of infected fish, including necrosis, degeneration, and structural damage.

1. IMPACT OF CLINOSTOMUM COMPLANATUM
INFECTION IN Nandus nandus WITH SPECIAL
REFERENCE TO HISTOLOGY
DISSERTATION
For the Partial Fulfillment of
Master of Fisheries Science
Department of Zoology and Applied Aquaculture,
Barkatullah University, Bhopal.
(2014)
SUBMITTED BY
DEEPENDRA SINGH RAJAWAT
UNDER THE GUIDANCE OF UNDER THE CO-GUIDANCE OF
Dr. Pradeep Shrivastava Dr. Pinky Kaur

2.  INTRODUCTION
 OBJECTIVE
 MATERIAL AND MEATHOD
 RESULT
 CONCLUSION

3. Aquaculture is the fastest growing food production sector in the
world and provides a significant supplement to, and substitute for,
wild fish and plants. However, disease has become a primary
constraint to aquaculture growth and is now responsible for the
svelopevere impact on both the economic and socio-economic
dement in many countries of the world. Addressing health questions
with both pro-active and reactive programmes has therefore become
an urgent requirement for sustaining the growth of aquatic animal
food production.
The effect of parasites on the fresh water fishes (Nandus
nandus) is of considerable importance because of its widespread
occurrence and an intermediate link in the food chain. Parasites may
effect host physiology in many ways that induce stress in the host.
Fish disease consitute one of the most important
problems and challenges confronting fish culturist.

4.  The presence of parasite Clinostomum complanatum in
fish Nandus nandus is infected the organ liver,
spleen, ovary and intestine. The parasitic cyst of
Clinostomum complanatum is found in liver and
spleen.

5.  Collection of the host fish :-
 The host fish Nandus nandus were collected from
local fish market of Bhopal regions like RRL,
Habibganj, Barkherra, Piplani, Govindpura market
for the collection of fish specimen. Length and
weight were also taken.

6.  The physical parameters like length, standard
length and weight of the fish were noted.
External examination of the fish includes through
examination of oral cavity, outer body surface,
fins and operculum which mostly harbour the
trematode cysts. The hosts brought to laboratory
were subjected to a through investigation as per
the methods employed by Cable, 1977 and Mayer
and Olson, 1975.

7. 1. Parasites were collected as ecto-parasites and endo-parasites
from skin, gills and internal organs by dissecting the fish,
respectively in normal saline and were examined alive condition
in order to identify them.
2. Parasites collected were fixed in fixative - alcohol formal acetic
(AFA) and glycerin alcohol in cuticular parasites (Humason,
1979).
3. The fixative should be thoroughly removed from specimens prior
to sustaining wash out formalin in distilled water and AFA in 70%
alcohol.
4. The specimens preserved in 70-90% alcohol were dehydrated
through the alcoholic series 30% (10mins), 50% (10 mins).
5. Then they were kept 5-10 minute in dilute Aceto Carmine for
staining.
6. Specimens were slightly washed within 70% alcohol stain.

8. 7. And further dehydration by passing specimens through an
accending series of alcohols i.e.70% (10 mins.), 90% (10 mins.)
and 100% alcohol(10mins.).
• After complete dehydration, the specimens were transferred to a
cleaning agent (Xylene) which rendered the specimens
transparent mount in DPX.
• The stains, dehydrated and cleared specimens were mounted in
Dextrin Plasticized Xylene (DPX) mountant.
• The cover glass was preferably lowered horizontally to avoid
trapping of air bubbles and displacement of specimens.
• Finally photographs were taken with the help of Olympus
Microscope at different combinations of magnification.

9. PREPARATION OF HISTOLOGICAL SLIDES :-
Diseased fish tissues were collected were quickly
washed with physiological saline solution (.75%NaCl) and
immediately fixed in 10% formalin ,dehydrated, paraffin
impregnation, paraffin embedding, blocking out of
impregnated tissuses, trimming and sectioning, affixing
and deparaffinization, rehydrated the tissues, stained in
hematoxyline dehydrated in alcoholic series and cleaned in
xyelene, mounted in DPX and labelled according to
samples.

10. (i) Prevalence = Total No. of Hosts Infected×100
Total No. of Hosts Examined
(ii) Mean Intensity = Total No. of parasites
Total No. of Infected Hosts Examined
(iii) Relative Abundance = Total No. Parasites
Total No. of Hosts Examined

11. S.No. MORPHOMETRICS CHARACTERS
1 Body Length 3.360-5.760
2 Body Width 1.20-2.60
3 Oral sucker (OS) Length 0.12-0.14
4 Oral sucker (OS) Width 0.24-0.88
5 Ventral sucker (VS) Length 0.56-0.72
6 Ventral sucker (VS) Width 0.24-0.72
7 Anterior Testis Length 0.52-0.54
8 Anterior Testis Width 0.50-0.51
9 Posterior Testis Length 0.42-0.45
10 Posterior Testis Width 0.78-0.79
11 Ovary Length 0.21-0.25
12 Ovary Width 0.3-0.32

12. 57 of Nandus nandus (ranging from 9-12 cm in total length) were
examines for helminth parasites. Of this number, 27 (47.36%) were found
to be infected by helminth parasites.
Total no. of fishes observed - 57
Total no. of fishes found infected - 27
Total no. of parasites collected - 138
Total prevalence% - 47.36
Total intensity - 5.11
Total abundance - 2.421
Present study revealed a total of 4 classes of parasitic helminthes were
found. These classes are the Cestodes, nematodes, trematodes and
acanthocephalan were collected during investigation.
The maximum prevalence was recorded in the month of November.
While the slightly less was observed in month of December and October.

13. The maximum prevalence was recorded in the month of November.
While the slightly less was observed in month of December and October

14. The maximum intensity was recorded in the
month of December. While the slightly less was
observed in month of October and minimum in
month of December

15. The histo-pathological effect of trematode in liver of N.
nandus, exhibited necrosis and loosening of hepatic
tissue, degeneration and shrinkage in hepatocytes and
shifting of nuclei at the periphery of cells.
The result showed the destructive and distartive results
in the liver, ovary and spleen when Nandus infected with
Clinostomum Complanatum.

16. Fig.6. Cross section of liver X100 Fig.7. Cross section of liver X400

17. Spleen :-
The red pulp which may occupy the majority of the organ is
containing too many sinusoids filled with red blood cells surrounded
by some trabeculae and diffused lymphatic tissues throughout the red
pulp of spleen. Infiltration by lymphoid cells and showing
hyperaemia.
Fig.8. Microphotography showing cross section of spleen X400

18. OVARIES :-
Histo-pathologically the ovaries of N. nandus exhibited
thining and damaged in their walls due to the association with
metacercariae. Pathological effect included irregular shape of
oocytes, and necrosis.

19.  The infection of Clinostomum complanatum in
Nandus nandus is occur in organ liver spleen
ovary where the parasitic cyst is found, it will
effect the tissues of these organ due to which
the metabolism of fish is distrubed .Thus it will
harmful for fish growth.

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