Livestock Research for Rural Development 27 (8) 2015 Guide for preparation of papers LRRD Newsletter

Citation of this paper

Seroprevalence study of swine brucellosis and knowledge of pig traders and dressers at slaughter facilities in Temeke Municipality of Dar es Salaam, Tanzania

C Simon, E D Karimuribo1 and B M Kessy2

Livestock Department - Temeke Municipal Council, P.O. Box 46343, Dar es Salaam, Tanzania
cleophasshayo@gmail.com
1 Department of Veterinary Medicine and Public Health, Faculty of Veterinary Medicine
Sokoine University of Agriculture, P. O. Box 3021, Morogoro, Tanzania.
2 Department of Veterinary Surgery and Theriogenology, Faculty of Veterinary Medicine, Sokoine University of Agriculture
P.O. Box 3020, Morogoro, Tanzania

Abstract

A cross sectional study was carried out between December 2013 and January 2014 to establish seroprevalence of swine brucellosis in pigs slaughtered at five pig slaughter facilities in Temeke Municipality located in Dar es Salaam city, Tanzania. The study also assessed knowledge and practices of pig traders, dressers and pork inspectors with respect to swine brucellosis. A total of 414 serum samples collected at five purposively selected slaughter facilities were screened using Buffered Plate Agglutination Test (BPAT) and Standard Plate Test (SPT). All positive reactors to BPAT and SPT were confirmed by Rivanol test. Overall, the seroprevalence of swine brucellosis was 0.7% (95% C.I=0.2-2.1) (3/414 pigs screened) based on Rivanol test results. Three positive reactor pigs recorded at Mchicha Tazara (1) and Keko A (2) pig slaughter facilities were traced to originate from Singida (2) and Simiyu (1) regions. It was further established that the majority of pigs slaughtered in Temeke originated from Manyara (36.0%), Dodoma (21.7%) and Singida (18.6%) regions. The majority of pig traders (66.7%) and pig dressers (94.4%) had limited knowledge on occurrence of swine brucellosis.

 

To the best of our knowledge this is a systematic study which has reported for the first time the seroprevalence of swine brucellosis in pig herds transported for slaughter in towns in Tanzania. Findings of this study indicate that swine brucellosis is prevalent albeit at low levels amongst slaughtered pigs. There is therefore a need to strengthen disease surveillance as well as adopting appropriate disease control measures in the country including sensitization of the public on the public health importance of swine brucellosis.

Key words: BPAT, Brucella, public health, Rivanol, SPT


Introduction

Brucellosis is a major zoonotic disease of public health affecting domestic animals, wildlife and humans (OIE 2009a). According to OIE, it is the second most important zoonotic disease in the world after rabies and it is common in countries that do not have good standardized and effective public health and domestic animal health programs (Kunda et al 2005; Abubakar et al 2011). The aetiological agent of brucellosis in pigs is Brucella suis (biovars 1, 2 or 3) which is a small non-motile coccobacillus or short rods facultative intracellular Gram-negative bacteria. Brucellosis caused by B. suis biovar 1 and 3 causes a severe disease in human similar to the one caused by B. mellitensis (Corbel 2006; Meirelles-Bartoli et al 2012; Rahman et al 2012). Pigs can get infection by consuming contaminated feed, water and during mating with infected pigs. Environment contaminated with reproductive organs discharges or aborted materials especially where different stock species are kept increases the risk of infection between pigs (Megid et al 2010).

 

Infection in pigs results in irregular oestrus, abortions, stillbirths, small litters and birth of weak piglets. Infected swine tissues have high concentration of B. suis organisms as compared with cattle (Meirelles-Bartoli et al 2012). This may have a serious implication in pigs and personnel dealing with pigs. Due to the long duration of human illness and its convalescence, brucellosis is perceived as an important socio-economic as well as a medical problem. Its clinical presentation in human confuse with other diseases like malaria and typhoid (Corbel 2006; Kunda et al 2007; WHO 2013).

 

Brucellosis can be controlled by test and slaughter. Diagnostic approaches include the use of widely acceptable serological screening tests like the Rose Bengal test (RBT) and the Buffered Plate Agglutination Test (BPAT). These tests use Brucella abortus antigen for serological testing of antibodies against all smooth Brucella species i.e. B. mellitensis, B. abortus, and B.suis due to cross reactivity caused by lipopolysaccharide or membrane proteins shared (Paulo et al 2000). It is recommended that positive reactor samples should be confirmed using an established confirmatory and/or complementary test such as Enzyme-Linked Immunosorbent Assay (ELISA), Fluorescence Polarisation Assay and Rivanol (OIE 2009b; Poester et al 2010; Montasser et al 2011). Culture remains the gold standard while DNA probes or polymerase chain reaction (PCR) methods can be used to demonstrate the agent in various biological samples.

 

In Tanzania pig keeping provides an opportunity as an income generating activity but is faced with a number of production constraints such as availability of quality feeds, prevalence of diseases like helminths and abortions (Karimuribo et al 2011). In recent years, there has been increased concentration of human population in urban and peri-urban areas (Kunda et al 2005). Urbanisation and population growth in Temeke has increased demand for food and quality protein source like pork. Since most pigs are kept in villages, pig traders purchase them from keepers, collect and pool them into one pen managed by pig trader temporarily (10-14 days) before being transported to Temeke where they fetch high price.

 

 There is scarcity of research information on the prevalence and factors associated with swine brucellosis in Tanzania. The present study aimed at establishing swine brucellosis seroprevalence and assess knowledge of pig traders and dressers at slaughter facilities in Temeke Municipality of Dar es Salaam, Tanzania. The study also attempted to document sources of pigs slaughtered in Temeke Dar es Salaam as well as practices of managing such pigs which might influence the occurrence of brucellosis in pigs. It was anticipated that findings from this study would help to guide further research and necessary intervention for improved pig production industry.


Materials and methods

Study area

 

The study was conducted in Temeke Municipal Council of Dar es Salaam city which lies between Latitude 6 48' and 7 10' South of equator, and between Longitude 39 33' and 40 East of Greenwich meridian. Temeke Municipality is composed of 30 administrative wards and 120 streets. It has total area of 786.5 km2 and was selected as a representative of Municipalities of Dar es Salaam city. Five out of eight purposively selected slaughter facilities in Temeke were used during this study.

 

Study design and sampling strategy

 

A cross sectional study was conducted between December 2013 and January 2014 to establish the magnitude of swine brucellosis in pig slaughter facilities in this survey. Pigs in the slaughter facilities were used as sources of pigs to be sampled. Each pig slaughter facility was visited once each time a new consignment of pigs arrived for slaughter until when required sample size was met. The maximum sample size was calculated according to formula by Naing et al (2006) [i.e.  n = Z2 p(1-p)/d2 ] where n= total number of pigs sampled to detect at least one infected animal at 95% confidence interval (Z statistic value =1.96), p= expected prevalence (p) of 50% at 5% precision (d). The prevalence of 50% (0.5) was used in this case to get maximum sample size as prevalence of the disease has not been established.

 

Purchased pigs from the pig keepers were identified by putting identification marks (shaved marks) right at the time of purchasing. Each pig was accompanied by information related to age, sex, identification mark, and management in the herd obtained through structured interview. Each time when new batches of pigs were received in the slaughter facilities holding pen, proportional contribution of each facility was calculated to establish the number of pigs to be picked for sampling from each facility. The identification marks of each pig present in respective slaughter facility were recorded in pieces of papers, folded, put in a box, mixed and each time a piece of paper was randomly picked without replacement. Respective pigs for each identification mark read from the randomly picked piece of paper were bled to get blood samples. A total of 414 samples were proportionally collected from the five pig slaughter facilities based on the established average number of pigs present in the holding pens as indicated in Table 1. 

Table 1: Number of pigs proportionally sampled from each slaughter facility holding pen

Facility name

Sex

Total number

Males

Females

Mchicha Tazara

70

102

172

Keko A

31

63

94

Keko B

24

52

76

Mama Jack and Nuru

14

9

23

Mbagala Charambe

18

31

49

Total

152

262

414

Structured interview to pig keepers, pig traders, dressers and pork inspectors

 

A questionnaire comprising of closed ended type questions was used to collect information from 115 purposively selected pig keepers who had pigs for selling to pig traders. Data collection approach employed face-to-face interview of pig keepers which was conducted by 21 pre-trained pig traders from the source regions of Pwani, Dodoma, Katavi, Manyara, Simiyu and Singida.  Individual pig information recorded were sex, age and the identification mark (either existing or marked by pig traders before being transported to Temeke-Dar es Salaam. Herd-related information recorded by pig traders included: herd size (small (≤ 10 pigs), medium (11-20 pigs) or large (≥ 21 pigs), feed sources (purchasing of readymade feed, buy ingredients and compound, use of green leafy material from the farm), pig management system (intensive or semi-intensive), history of buying new pigs (within the past one year), sharing of boars, contact with wild pigs, presence of other livestock species in the farm, reproductive health issues like abortion and still birth, handling and disposal of placenta or aborted foetuses. Data collection from 21 pig traders, 19 dressers and seven pork inspectors was carried out by the researcher (CS) using face-to-face interviews. A close-ended questionnaire was used to collect information on knowledge and practices in respect to risks and preventive measures taken by pig slaughter facility personnel against swine brucellosis.

 

Blood sampling at slaughter facilities

 

Collection of blood samples from 414 pigs was carried out by the researcher at each slaughter facility that participated in the study. By the use of a vacutainer needle holder and tube, five millilitres (ml) of blood were aseptically collected from the pig’s vena cava using plain vacutainer tubes (Vacuum tube sterile, Zhejiang Kangsh Beijing). Each serum sample was clearly labeled and left in standing position in a rack within a cool box during sample collection in the field. The collected blood samples were taken to Temeke Municipal Livestock Office the same day for centrifugation with an automatic electric powered centrifuge at a speed of 2500 rotations per minute for 10 minutes to get clear serum. The harvested serum samples were pippeted into sterile cryovials, labeled and stored in a refrigerator at 4C for a maximum of seven days after which all sera were shipped to Sokoine University of Agriculture (SUA) laboratory in a cool box with ice packs where they were stored at -20C until analysis.

 

Buffered Plate Agglutination Test (BPAT), Standard Plate Test (SPT) and Rivanol (RivT) tests

 

All 414 serum samples collected were screened for swine brucellosis using BPAT according to procedures by OIE (2009a). The antigen used was Brucella abortus kindly supplied by the National Veterinary Services Laboratory (NVSL- USA -NVSL 1920 serial number 10-51303 Dayton Avenue Ames IA 50010). The test was performed on a glass plate where 80 microlitre (l) of serum samples, positive and negative control were pipetted followed by additional of 30l of antigen. The antigen and sera were hand mixed using a pipette tip in circular manner. The plate was hand rocked twice clockwise and anticlockwise at intervals of four minutes and incubated at room temperature for four minutes. Results were read after eight minutes in a bright light guided by the controls. Any agglutination was recorded as a positive reaction and negative if there was no agglutination.

 

Standard Plate Test (SPT) was performed according to Alton et al (1975) on each BPAT positive serum sample. Briefly, from each BPAT positive sera and controls, decreasing quantities i.e. 80l, 40l, 20l and 10l were pipetted onto a glass plate. Thirty microlitre of antigen (NVSL 1920 serial number1-1301, Dayton avenue USA) were placed adjacent to each sera and mixed. The plate was then rocked four times in between four minutes interval and results were recorded after eight minutes.

 

Confirmation of the positive samples was done using Rivanol test (RivT) which was performed according to the reagent manufacturer instructions (NVSL Dayton Avenue-USA). Briefly 300l each positive serum sample and controls were pipetted in tubes into which equal volume of rivanol solution were dispensed, mixed and left at room temperature for 5 minutes. The mixture was centrifuged at 1000 rotations per minute (rpm) for five minutes. From the supernatant 80l, 40l, 20l and 10l were placed onto a glass plate and 30 l of antigen were added to each and mixed starting from the lowest concentration (10) to the highest (80l). The glass plate was hand rocked four times in between six minutes interval and results read after 12 minutes. Any agglutination was recorded as a positive reaction and negative if no agglutination against their dilution. The cut-off point for positive sample was 1:64

 

Data analysis

 

All data were entered and cleared in Microsoft Office TM 2007 Excel sheet and later exported to EPI info TM Version 7.1.1.14 (Centre for Disease Control, Georgia Atlanta USA 2007) for analysis. Descriptive and inferential statistics of the outcome variable were calculated as proportions. The proportions were compared by Chi square test at 95% confidence level to establish any statistical significance.


Results

Characteristics of pigs slaughtered in Temeke

 

It was found that 103 (89.6%) out 115 pig keepers had small herd size. The herd size ranged from one to 104 pigs. Most pigs were from Manyara, followed by Dodoma, Singida, Katavi and Pwani regions with their percentage contribution as indicated in Figure 1.

Figure 1. Source areas for the pigs transported for slaughter

The majority of pigs purchased by pig traders were females which overall constituted 63.3% of all 414 pigs for slaughter as shown in Table 2. This trend of buying more female pigs than boars was practised similarly by pig traders in all six regions where the proportion of female pigs ranged from 58.4% (Manyara region) to 88.0% (Pwani region). Most pigs purchased were of adult size constituting 73.3% of all pigs transported to Temeke Municipality. With exception to Mama Jack and Nuru facility (which received pigs from Simiyu only), other slaughter facilities received slaughter pigs from more than one region (three to four regions per facility) as indicated in Table 2.

Table 2: Main characteristics of pigs purchased for slaughter in Temeke

Variable

Level

Source region

Pwani

Dodoma

Katavi

Manyara

Simiyu

Singida

Total

Percentage (%)

Sex

Male

3

33

6

62

20

28

152

36.7

Female

22

57

18

87

29

49

262

63.3

Age group

Adults (= 10 months)

19

70

23

114

28

48

302

72.9

Finisher (< 10 months)

6

20

1

35

21

29

112

27.0

Small (= 10 pigs)

6

32

8

41

0

16

103

89.6

Herd size

Medium (11-20 pigs)

0

0

0

3

6

2

11

9.6

Large (= 21 pigs)

1

0

0

0

0

0

1

0.9

Management
system

Intensive (Pen)

7

32

8

44

1

18

110

95.7

Semi intensive

0

0

0

0

5

0

5

4.4

Purchasing of
new pigs*

Yes

3

22

3

38

6

12

84

73

No

4

10

5

6

0

6

31

27

Sharing of boars

Yes

6

31

7

43

6

13

106

92.2

No

1

1

1

1

0

5

9

7.8

Slaughter facility

Mchicha Tazara

6

60

0

80

26

0

172

41.6

Keko A

17

0

17

17

0

43

94

22.7

Keko B

2

18

7

49

0

0

76

18.4

Mama Jack and Nuru

0

0

0

0

23

0

23

5.6

Mbagala charambe

0

12

0

3

0

34

49

11.8

* Within the last 12 months

Pig management and reproductive health at source villages

 

The majority (53.9%) of pig keepers reported to buy readymade feed for their pigs. Boar sharing was reported by 106 (92.2%) out of 115 pig keepers interviewed. Twenty (17.4%) out of 115  pig keepers reported cases of abortion in their pigs while still birth was mentioned by seven (6.1%) pig owners. History of abortion was high in Manyara (seven cases), followed by Dodoma (five), Simiyu (4) and Singida (3) as shown in Table 3. Sixty one (53.04%) of 115 pig keepers reported to use bare hand when disposing the placenta following furrowing/abortion, 31 (26%) use a hoe while 23 (20%) used a spade.

Table 3: Occurrence of pig reproductive health problems as reported by pig keepers

Variable

Level

Region

Total
number

Percentage
(%)

Pwani

Dodoma

Katavi

Manyara

Simiyu

Singida

History of abortion

Yes

0

5

1

7

4

3

20

17.4

No

7

27

7

37

2

15

95

82.6

Abortion events frequency

1

0

5

1

3

4

2

15

13

2

0

0

0

4

0

1

5

4.4

History of still birth

Yes

0

1

0

5

0

1

7

6.0

No

7

31

8

39

6

17

108

93.9

Knowledge and practices of pig traders, dressers and pork inspectors

 

Out of 19 pig dressers interviewed, 15 (79.0%) belonged to age group of 21-40 years. The majority had working experience of 4-6 years in pig dressing. Eighteen out of 19 pig dressers (94.7%) had limited knowledge on disease they could get from dressing pigs. Four out of 10 who reported to know the disease were not aware of the protective measure to be taken and wound were the major health problem reported by 14 (73.7%) of 19 pig dressers interviewed as shown in Table 4.

Table 4: Summary of questionnaire administered to pig dressers in Temeke

Parameter

Level

No. of Respondents (n=19)

Percentage (%)

Pig dressers age

21-40 years

15

79

41-60 years

4

21

Number of years worked

Less than 1 year

2

10.5

1-3 years

4

21.5

4-6 years

9

47.4

7-9 years

1

5.3

Over 10 years

3

15.8

Knowledge of health problems associated with pig dressing

Yes

15

79

No

4

22

List health problems associated with pig dressing

Wounds

14

73.7

Dont know

3

15.8

Wounds and diseases

2

10.5

Knowledge of diseases they can get from pigs

Yes

1

5.3

No

18

94.7

Ever heard of swine brucellosis

Yes

10

52.6

No

9

47.4

Source of information about swine brucellosis

Fellow in work

4

40

I saw it in my farm

3

30

Neighbours

3

30

Ways to protect from the disease

Dont know

4

40

Wear protective gears

6

60

Use of reproductive organs from slaughtered pigs

Sold to customers

19

100

Offered to people

0

0

All seven (100.0%) pork inspectors were aware of diseases they could contract from pigs. They were aware of transmission of swine brucellosis, the symptoms and protection methods from the disease. All 21 pig traders interviewed mentioned that, all the pigs transported were for slaughter. All traders mentioned to pool collected pigs from different pens into one. Fourteen (66.7%) out of 21pig traders were not aware of disease they could contract from pigs. It was further found that even the pig traders (52.2%  n=11)   who mentioned to ever heard of swine brucellosis, were not aware if the disease affects human beings and nine out of these (81.8%) didn’t know the protective measure to be taken as shown  in Table 5.

Table 5: Summary of a questionnaire administered to Pig traders

Parameter

Level

No. of respondents (n=21)

Percentage (%)

Pig traders age

21-40 years

12

57.1

41-60 years

9

42.9

Number of years of transporting pigs

Less than I year

6

28.6

1-3 years

5

23.8

4-6 years

8

38.1

7-9 years

1

4.7

Over 10 years

1

4.7

Purpose of pigs transported to Temeke

Slaughter

21

100

Sell to pig keepers

0

0

Weeks taken to collect pigs for transportation

One week

4

19.1

2 weeks

14

66.7

Over 2 weeks

3

14.3

Weeks taken for brought pigs to get slaughtered

One week

5

23.8

2 weeks

16

76.2

Over 2 weeks

0

0

Knowing diseases they can get from pigs

Yes

7

33.3

No

14

66.7

Ever heard of swine brucellosis

Yes

11

52.4

No

10

47.6

Does swine brucellosis affect human

Yes

0

0

No

11

100

Ways to get protected from the disease

Wear protective gears

2

18.2

Dont know

9

81.8

Seroprevalence of swine brucellosis

 

The seroprevalence of 0.7% (95%, C.I=0.2-2.1 i.e. three out of 414 pigs screened), was recorded in this study. All three seropositive animals belonged to the adult category of pigs (≥10 months). Of the three seropositive pigs, two were females and one was a boar. The two females were sampled from Keko A pig slaughter facility site while one boar was from Mchicha Tazara slaughter facility giving slaughter site seroprevalence of 2.1% and 0.6% respectively. They were originating from Singida (2) and Simiyu (1) regions respectively. 


Discussion

Three pigs were commonly found in most pig herds. The minimum, average and maximum age for pigs slaughtered was six, 12.4 and 28 months respectively. The majority of purchased pigs were females, probably pig keepers kept females than males for breeding purposes. The adult pigs dominated likely because at an average of 12 months most pigs reach appreciable size for selling by most small scale pig keepers. Most pigs were from small scale pig keepers probably due to low capital required for investing in small scale pig production. Pig traders purchased pigs from Manyara, Dodoma, Singida, Simiyu and Pwani. Sharing boars for mating sows on heat and purchasing of new pigs within a year was practiced by most pig keepers probably because female pigs were more kept than boars for breeding, but also sold pigs need to be replaced by purchasing new ones so as to remain in production. Basing on reproductive health problems, single abortion events were highly reported in Dodoma, followed by Simiyu and Singida. Improper disposal and handling of aborted material/foetuses reported were indication that pig keepers were not aware of the risk associated with handling the placenta with unprotected hands, as well as contamination of the environment which may transmit infection to other livestock in farms.

 

Most pig dressers were not aware of swine brucellosis and preventive measures due to limited information about the disease. The use of reproductive organs (tests, uterus and mammary glands commonly known as “lipsi”) reported by pig dressers is a health risk to customers during purchasing and to consumers since Brucella predilect in these organs (Corbel 2006).  Although about half of the pig traders reported to have ever heard of swine brucellosis they were not aware if the disease affects human beings. Cysticercosis was only the disease known by pig traders which can be explained by experience of condemned carcasses during post mortem inspection. All (seven) pork inspectors were aware of swine brucellosis due to the training they undergo in colleges.

 

Individual seroprevalence findings of the present study were slightly higher than those found in Nigeria by Onunkwo et al (2011) who reported a seroprevalence of 0.6% in pigs. However other studies in Croatia (Cvetnic et al 2009) and in Greece (Burriel et al 2003) reported higher seroprevalence recorded at 1% and 3% respectively. Studies by Van der Giessen and Priadi (1988) in Jakarta and Surabaya abattoirs in India, documented even the highest seroprevalence of 22.3% and 14.9%, respectively. The present study recorded swine brucellosis in only sexually matured pigs which could be due to increased time for exposure with age and reproductive organs maturity where Brucella organisms predilect (Corbel 2006).


Conclusions


Acknowledgement

The authors would like express their sincere thanks to the African Development Bank (ADB) through the South African Development Countries Transboundary Animal Diseases (SADC-TADs) project secretariat for funding this work. We are gratefully to the Ministry of Livestock Development and Fisheries, Faculty of Veterinary Medicine (SUA), Temeke Municipal Council Director, pig owners and pig market stakeholders for enabling this research to be accomplished.


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Received 8 June 2015; Accepted 1 July 2015; Published 1 August 2015

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