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Seroprevalence of Antibodies to Toxoplasma gondii in Butchers and Buffaloes at Lahore Pakistan.

Byline: Awais Anees Azhar Maqbool Umbreen Javed Khan Ghazalah Yasmin and Farhat Zahra

Abstract

The purpose of this study was to record the seroprevalence of Toxoplasma gondii antibodies in buthchers and buffaloes at Lahore Pakistan. Blood samples of 50 butchers and 50 buffaloes were taken from main abattoir and local buffalo farms around Lahore and assessed for seroprevalence of T. gondii by using a latex agglutination test (LAT). In the present study 10% of butchers and 14% buffaloes were found seropositive for T. gondii infection. Butchers ranging in age from 51-60 (n=12) years and above exhibited the highest seropositivity 25% followed by those aging 41- 50 years (n=10) 20%. Similarly in buffaloes 14% was also related to age by using LAT.

Key words: Toxoplasmosis seroprevalence butcher buffaloes.

Toxoplasma gondii is an intracellular protozoan parasite (Dubey 2009) which infects humans as well as a wide variety of mammals and birds Tenter et al. (2000) Sadia et al. (2012). It exhibits a predator-prey type life cycle and felines are the only definitive host (Tenter and Oswaldo 2009). T. gondii is found throughout the world and tends to be more prevalent in tropical climates. Toxoplasma has a complex life cycle consisting of intestinal and tissue phases. The intestinal phase of the infection occurs only in felines and exhibits a typical intestinal coccidian life cycle consisting of merogony and gamogony. The tissue phase of the infection can be transmitted congenitally to offspring and to other intermediate hosts through carnivores (Al-Daoody 2012). Ingestion of an infected animal will release the bradyzoites from the tissue cysts which then infect cells in the new host. The infection is acquired from under cooked meat.

During the 1980s' toxoplasmic encephalitis emerged as a common complication associated with AIDS (Marques et al. 2008; Sukthana 2006; Jones et al. 1999; Parmley et al. 1992). Congenital toxoplasmosis is more like to be symptomatic and can be particularly severe. The parasite can be transmitted to humans in several ways including ingestion of raw or undercooked meat contact with soil eating poorly washed raw fruits and vegetable and ingestion of oocysts in cat faeces (Berger- Schoch et al. 2011). An investigation of recent outbreak has suggested that T. gondii could also be waterborne (Zahida et al. 2013; Jones and Dubey 2010; Heukelbach et al. 2007; Dubey 2004).

Due to the increasing risk of public health by ingestion of contaminated meat and milk the development of reliable and sensitive method for diagnosing the toxoplasmosis becomes very important. As the butchers are considered to be at more risk for toxoplasmois and buffaloes are important source of meat in Pakistan the aim of present study was to record the seroprevalence of toxoplasmosis in butchers and buffaloes in Lahore Pakistan.

Materials and methods

A total of 100 serum samples (butchers n=50 and buffaloes n=50) were analyzed for anti- Toxoplasma antibodies. These samples were collected randomly from Lahore abattoir and local buffaloes farms. History of butchers and buffaloes was noted. Under aseptic measures 8-10 mL of blood was drawn (intravenous) with the help of disposable syringes and transferred slowly to a screw capped sterile clean test tube to avoid hemolysis (Figueiredo et al. 2001).

The samples were left for about an hour for blood clotting to occur. The clotted blood was then separated with a fine loop and serum samples were centrifuged at 3500 rpm for 5 minutes. The sterile serums supernatants were stored at -20C until further analysis. The serum samples were analyzed for specific IgG Toxoplasma antibodies using Latex Agglutination (LA) commercial test kit "Toxoplasma Latex" from Quimica Clinica ApelicadaSA Amosta Spain. The "Toxoplasma gondii Latex" was used according to manufacturer's instructions.

Results

Table I shows the prevalence of anti-T. gondii antibodies in sera collected from five age groups of butchers by latex agglutination test (LAT). Amongst the five groups of butchers Group-5 (51-60 years and above) had the highest seropositive percentage i.e. 25% followed by Group-4. The rest of the age groups had no positive sample (Table I). Two butchers showed antibody titer at screening dilution of 1:16 two butchers gave antibody titer of 1:128 and one was positive at screening dilution of 1:256 (Table I). Overall seropositive percentage by using LAT was 10% in butchers at Lahore Abattoir.

Table I.- Prevalence of anti-Toxoplasma gondii antibodies in sera collected from different age groups of butchers and

###buffaloes by latex agglutination test (LAT).

Age (Years)###No. of###Antibody titer###Sero positive###Sero positive

###individual###1:64###1:128###1:256###(%)###(%)

Butchers

Group-1 (15-20)###04###0###0###0###0###0

Group-2 (21-30)###06###0###0###0###0###0

Group-3 (31-40)###08###0###0###0###0###0

Group-4 (41-50)###10###1###1###0###2###20

Group-5 (51-60) and above###12###1###1###1###3###25

Total###50###02###02###01###05###10

Buffaloes

Group-1(1-5)###10###0###0###0###0###0

Group-2 (5-10)###12###1###1###0###2###16.66

Group-3 (10-15)###28###1###2###2###5###17.85

Total###50###2###3###2###7###14

Table I shows the prevalence of anti-T. gondii antibodies in sera collected from three age groups of buffaloes by LAT. Buffaloes in Group-3 (10-15 years and above) had the highest seropositive percentage (17.85%) Group-2 (5-10 years) had 16.66% seropositive while the Group-1 (1-5 years) had no positive case. Two buffaloes showed antibody titer at screening dilution of 1:16 three buffaloes showed antibody titer at 1:128 and two showed antibody titer at screening dilution of 1:256 (Table II). Overall seropositive percentage in buffaloes for Toxoplasma antibodies was 14%. History of butchers and buffaloes was noted (Table II). The data collected was analyzed and presented in terms of percentage so that prevalence of T. gondii antibodies and their relation with cats can be proved. There exist a positive correlation between the animal (cats) contact with the butchers and buffaloes (Tables II).

Discussion

In the present study the overall prevalence of anti-toxoplasma antibodies in butcher was 10%. Among 50 butchers examined in the present study an antibody titer of 1:256 was found in one butcher which suggested the possible recent contact an antibody titer of 1:128 was found in two butchers which was due to acquired or evolving immunity an antibody titer of 1:16 was also found in two butchers which indicated residual or non-specific immunity. In general low titers indicated past exposure and probable immunity while high titer of 1:256. These results are in agreement with Chaudhry et al. (2006) who randomly collected fifty serum samples from butchers for analysis of T. gondii by LAT. Out of which14 samples showed at 1:16 dilution. Out of them 10 were seropositive on 1:64 dilutions. It shows 20% prevalence of the toxoplasmosis in butchers.

Table II.- Animal Information and butchers information.

###n###Response###%

###frequency

Buffalo

Group-1 (1-5)###50###10###20

Group-2 (5-10)###50###12###24

Group-3 (10-15)###50###28###56

Sex (all female)###50###50###100

Breed-Cross breed (Neeli and###50###50###100

Ravi)

Contact with cats###50###50###100

Butchers

Age Group

Group 1 (15-20)###4###8

Group 2 (21-30)###6###12

Group 3 (31-40)###8###16

Group 4 (41-50)###10###20

Group 5 (51 and above)###22###44

Educational achievement

Primary###30###60

Secondary###16###32

Tertiary###4###8

Year of service###(Average)

15###3###6

610###5###10

1120###7###14

2130###10###20

greater than 30###25###50

Type of training

Formal training###7###14

On-the-job training###43###86

Safety information shared

Keep clothes clean###Yes###8(12)

Keep surroundings clean###No###43(86)

Wash hands###No###0(0)

Wear protective gears###No###1(2)

Other/Don't remember###NA###0(0)

Begin work day with clean

protective clothing

Always###No###45(90)

Sometimes###Yes###5(10)

Last medical exam

Current year###2###2(4)

1 year before###6###6(12)

2 years before###8###8(16)

3 years before###5###5(10)

4 years before###19###19(38)

Can't remember/No response###10###10(20)

Similar results were reported by Mazumder et al. (1988) and Samad et al. (1997) who used LAT for the detection of antibodies against T.gondii in butchers and found 24% and 50% cases positive respectively. Zaki (1995) also supported the results of present report by studying seroprevalence of T. gondii in domestic animals whereas findings of Zhang and Wei (2000) were in partial favor with the results of present report who evaluated the diagnostic value of LAT in 288 human serum samples for the detection of antibodies of T. gondii and found 8.9% seropositivity with positive titers (greater than or=1:32)

As far as the age of butchers and anti-toxoplasma antibodies titer relationship is concerned the highest percentage seropositivity was observed in Group-5 followed by Group-4. This might be due to the continuous exposure to the contaminated environment at abattoir and livestock farms and availability of the final hosts such as cats. These findings are in agreement with the results of Chaudhari et al. (2006) and Samad et al. (1997). The older butchers had higher serpositive cases for anti-Toxoplasma gondii antibodies than younger butchers this statement supports the study conducted by Jones et al. (2009) Pawelec et al. (2002) and Young (2000).

It was also noted that those butchers who had been in close and prolonged contact with the animals at abattoir showed highest seropositive results of toxoplasmosis. These findings are in agreement with those of Fayomi et al. (1987) who had shown the highest prevalence of toxoplasmosis (87.0%) among the staff of slaughterhouse. Overall prevalence of anti-toxoplasma antibodies in buffaloes was recorded as 12%. Prevalence of anti- toxoplasma antibodies were studied by researcher around such as El-Ridi et al. (1990) in Egypt (20.0%) Samad et al. (1997) in Bangladesh (12.09%) and Zaki (1995) in Pakistan (25%). This variation in the results could be due to difference in environmental and geographical conditions different ways of consuming meat like undercooked uncooked and unhygienic conditions e.g. contamination of food items by flies and other insects carrying oocysts and intimate association with pets (Al-Daoody 2012; Edelhofer and Prossinger 2010; Torada 2001; Wallace 1972).

The seroprevalence of T. gondii in buffaloes varied with different age groups. The highest seropositivity of (17.85%) was found in Group 3 (10-15 years and above) followed by Group -2 (16.66%) and no seropositive case was recorded in Group-1 (1-5 years). The present study revealed that the prevalence of antibodies increased with age. These findings are in concomitant with (Santos et al. 2010 2013; Figueiredo et al. 2001) that infection was higher in older animals than younger animals.

Among 50 buffaloes examined in the present study anti- body titer of 1:16 was recorded in two buffaloes who indicated residual or nonspecific immunity 3 showed antibody titer of 1:128 which was due to acquired or evolving immunity whereas two buffalo were positive at antibody titer of 1:256 which suggested no recent contact with parasite.

From the data it was concluded that seroprevalence of toxoplasmosis in buffaloes was low (14.0%) in Lahore Pakistan. Clinical disease is not unusual in immunocompromised individuals and ruminants. The prevalence of Toxoplasma infection in humans and animals is often associated with infection in pets (Al-Daoody 2012; Edelhofer and Prossinger 2010). Toxoplasmosis is zoonotic infection occurring in man in domestic animals wild animals and birds. Keeping in view of these findings proper system of management and health practices should be adapted to reduce the exposure rates.

Acknowledgement

All praises and thanks to Almighty Allah who taught man who he did not know. The literature of subject was provided to me by the Department of Zoology Punjab University Lahore; University of Veterinary and Animal Sciences Lahore and the Pakistan Scientific and Technical Information Center (PASTIC) Islamabad. Contribution of all these institutions of national importance in the field of science and research is highly appreciated.

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Author:Zahra, Farhat; Yasmin, Ghazalah; Khan, Javed; Umbreen, Maqbool; Azhar, Awais Anees
Publication:Pakistan Journal of Zoology
Article Type:Report
Geographic Code:9PAKI
Date:Oct 31, 2014
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