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Gut Contents of Cockroach PDF

This document describes a study examining the gut contents of cockroaches for protozoa. It outlines the materials and reagents used, including saline solution, fixatives, stains, ethanol, xylene and glassware. The procedure involves dissecting the gut, making a smear, fixing, staining and observing under a microscope. Various protozoa were observed, including Gregarina sp., Nyctotherus sp. and Ballantidium sp. Their systematic positions and identifying characteristics are provided.

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Debmalya Biswas
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2K views5 pages

Gut Contents of Cockroach PDF

This document describes a study examining the gut contents of cockroaches for protozoa. It outlines the materials and reagents used, including saline solution, fixatives, stains, ethanol, xylene and glassware. The procedure involves dissecting the gut, making a smear, fixing, staining and observing under a microscope. Various protozoa were observed, including Gregarina sp., Nyctotherus sp. and Ballantidium sp. Their systematic positions and identifying characteristics are provided.

Uploaded by

Debmalya Biswas
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Study of gut

contents of
cockroach for
protozoa
B.Sc. Part – III(Honours)

Prof. Jyoti Das


8/25/2017
Aim: To study the gut content of cockroach for protozoa
Materials Required:
a) Sample: Adult cockroach (Periplaneta americana)
b) Reagents:
i. 0.5% NaCl solution ,
ii. Schaudinn’s fixative,
iii. Iodinated alcohol,
iv. Distilled water,
v. Heidenhain’s haematoxylin,
vi. 3% iron alum,
vii. 1% iron alum,
viii. Ascending and descending grades of ethanol,
ix. Xylene,
x. DPX.
c) Glasswares:
i. Glass dropper/ Pasteur pipette,
ii. 100 ml measuring cylinder,
iii. 50 ml and 100 ml beaker,
iv. Petridish with cover,
v. Glass rod,
vi. Dropping bottle,
vii. Coupling jar,
viii. Grease free glass slides,
ix. Cover slips.
Preparation of Reagents:
Reagents Components Method of preparation
i. 0.5% NaCl solution  NaCl – 0.5g,, 0.5g of NaCl wax
 Distilled water – dissolved in 100ml of
100ml. distilled water and mixed
thoroughly.
ii. Schaudinn’s fixative  Saturated aqueous The saturated aq. Solution
mercuric chloride of mercuric chloride was
solution (2 parts), mixed with ethanol in the
 Absolute ethanol (1 ratio 2:1. It was mixed
parts). thoroughly and kept in a
 Glacial acetic acid glass container covered.
iii. Iodinated alcohol  A few crystals of The two reagents were
iodine, mixed and kept in a
 70% ethanol – coupling jar.
50ml.
iv. Heidenhain’s  Haematoxylin The haematoxylin powder
haematoxylin (mordant powder – 0.5g, was dissolved in 10 ml
free)  Distilled water – 95% ethanol and then
100ml, distilled water was added.
 95% ethanol – The stain was then ready
10ml. for use.
v. 3% iron alum  Ferric ammonium 3gm of Ferric ammonium
(mordant) sulphate – 3gm, sulphate was dissolved in
 Distilled water – 100ml distilled water and
100ml. stirred and kept as a
coupling jar.
vi. 1% iron alum  Ferric ammonium 1gm of Ferric ammonium
(differentiating reagent) sulphate – 3gm, sulphate was dissolved in
 Distilled water – 100ml distilled water and
100ml. stirred and kept as a
coupling jar.

Procedure:
a) Dissection of the sample:
The sample was taken and pinned down on a dissection tray. It was
cut open longitudinally and the hindgut was smeared and collected in
a watch glass.
b) Smear Preparation:
The smear of the gut content was made on a very thin, grease free
slide with the help of 0.5% NaCl solution.
c) Fixation:
The smear was then fixed in Schaudinn’s fixative for 10-15 minutes.
d) Staining:
i. The slide was dipped in iodinated alcohol for 1 minute.
ii. The slide was passed through descending grades of ethanol
70%, 50%, 30% each for 1 minute.
iii. Next, it was dipped in distilled water.
iv. The smear was then stained in Heidenhain’s haematoxylin
followed by a dip in distilled water. It was then observed under
a microscope.
v. If proper staining has taken place, the slide was dipped in 1%
iron alum for differentiation.
vi. After proper differentiation the slide was rinsed in distilled
water and then dehydrated by passing through ascending grades
of alcohol for 2-3 min each.
vii. It was then quickly passed through xylene and thereafter the
slide was mounted in DPX and observed under microscope.
Observation:
Various parasites were observed in the smear prepared from the gut content of
cockroach. The observed parasites are as follows:
a) Gregarina sp.
Systematic Position:
Subkingdom – Protozoa
Phylum – Apicomplexa
Class – Sporozoea
Subclass – Gregarina
Genus – Gregarina

b) Nyctotherus sp.
Systematic Position:
Subkingdom – Protozoa
Phylum – Ciliophora
Class – Polymenophora
Subclass – Spirotrichia
Order – Heterotrichida
Genus – Nyctotherus
Identifying Characters:
 Almost oval shaped body.
 Presence of a large irregular macronucleus and a short
micronucleus.
 Cytopharynx well developed.
 Cytopyge at the posterior end.

c) Ballantidium sp.
Systematic Position:
Subkingdom – Protozoa
Phylum – Ciliophora
Class – Litostomatea
Order – Vestibuliferida
Genus – Ballantidium
Identifying Characters:
 Outer lining covered by longitudinal row of spial cilia.
 Vestibule at the anterior end that leads to cytosome.
 Presence of food vacuole and contractile vacuole.
 Presence of cytopyge at the anterior end.

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