r&d (farah)

7/28/2019 R&D (Farah)

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SUPERVISORS:

DR. SITI NURSHEENA

PROF. G. SURESH KUMAR

DR. P. CHANDRAWATHANI

FARAH HAZIQAH BINTI MEOR TERMIZI

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INTRODUCTION

Blastocystis

- single-celled protozoan parasite

- found in the gastrointestinal tract

- B. hominis (in humans)

- Blastocystis sp. (other animals hosts)

- 4 common forms

(cysts, vacuolar, granular & amoeboid)

- fecal-oral transmission

(involves only the cyst form) Figure 1: Life cycle of Blastocystis

Tan (2004)

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Zoonotic infection

- Blastocystis is considered to be azoonoses

- Numerous Blastocystis isolatesfrom humans are believed to bepotentially zoonotic

: they have similar genotypes to

isolates found in a variety of otheranimals and birds species

Figure 2: Life cycle for Blastocystis cells

suggesting the existence of zoonotic

genotypes (subtypes 1 - 7) with various

host specificities (Tan, 2008).

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Lack of data

- most studies concentrated on Blastocystis inhumans

- Blastocystis sp. infections appear to be common in

birds

Japan: domestic fowls and ostriches 100% infection (Yamada et al., 1987a)

Russia: domestic hens 80 – 100% infection (Belova and Kostenko, 1990)

Czech Republic: domestic ducks 80% infection (Pakandl and Pecka, 1992)

- limited studies of Blastocystis from chickens/poultry inMalaysia

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OBJECTIVES

1) To determine the prevalence and morphology of Blastocystis sp. in chickens

2) To assess the in vitro growth pattern of Blastocystis sp. isolated from chickens (in progress)

3) To identify Blastocystis sp. subtypes from chickenisolates by PCR (in progress)

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MATERIALS & METHODS• Feb - May 2013: 68 chickens

• sampling sites- Tronoh wet market

- Titi Gantung wet market

- Super Kinta wet market

- Chemor pet shop

- Batu Gajah pet shop

- Chepor backyard chicken farm

- Semenyih chicken farm (Selangor)

• convenience sampling method - depends on availability

•

5 types of chickens:i. ‘ayam kampung’ (Village chickens)

ii. ‘ayam hutan’ (Jungle fowl)

iii. ‘ayam pedaging’ (Broiler chickens)

iv. ‘ayam kacukan’ (Crossbred chickens)

v. ‘ayam selasih’ (White silkie chickens)

‘ayam kampung’

‘ayam hutan’

‘ayam kacukan’

‘ayam selasih’

‘ayam pedaging’

(Perak)

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Faecal sample collection

the sample should be as fresh as possible

sample was stored in sterile stool collection container

Figure 2: Faecal sampling

(Photo by Farah)

Figure 3: Stool collection container

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Jones Medium (Standard medium)

In Vitro cultivation

Examine faecal suspension Incubate 370C; 48-72 hours

A pea size of fecal + 3 ml medium

(Photo by Farah)

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Stain with

Giemsa

Examine

under microscope

Fix with

methanol

Prepare a

thin smear

Slide preparation

(Photo by Farah)

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RESULTS & DISCUSSION

Table 1: Prevalence of Blastocystis sp. in chickens.

Study animals No. of faecal samples No. of chicken infected (%) Caged chickens

"Ayam pedaging" 15 0 Free-range chickens

"Ayam hutan" 1 1 (100) "Ayam kampung" 72 24 (33.3)

"Ayam selasih" 2 2 (100) Total 90 27 (30)

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Backyard chicken/free-ranged chickens ~ highprevalence

: scavenging habits

: unhygienic area

: pick up the infection from environment

contaminated with the faecal material of animals Cage-reared chicken ~ free of infection

: well managed, hygienic

: treated with antibiotic, vaccination

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Figure 4: Vacuolated form (arrow) of Blastocystis sp. from

faeces of chickens (x400).

(Photo by Farah)

• The vacuolar (or vacuolated form) was the most common Blastocystis cellform found in culture.• Vacuolar cells are usually spherical.• Cells vary greatly in size, ranging from 10µm to 30µm in diameter.

Figure 5: Vacuolar forms showing different size of Blastocystis

at 1000x magnification. (CV: central vacuole)

CV

CV

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Figure 6: Binary fission (BF) of vacuolated form in culture (x1000).

(Photo by Farah)

BF

• There are various reproductive mode of Blastocystis.• In cultures, it showed that the cell division with binary fission is the mainreproductive stage.

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Humans contract Blastocystis infection by drinking water/eating food contaminated with faeces from the infected animals.

common signs/symptoms :

- abdominal pain - constipation

- diarrhea - weight loss

- fatigue - flatulence

some may not have symptoms

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Prevent Blastocystis infection

1) Practice good hygiene

2) Avoid water/food that might be contaminated

- do not drink untreated water

- wash all raw vegetables/fruits before eating- avoid eating uncooked /undercooked food

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THANK YOU

r&d (farah)

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