Comparison of Cape Town and Skirrow's Campylobacter isolation ...

Trop Anim Health Prod (2011) 43:1007–1013 DOI 10.1007/s11250-011-9799-z

ORIGINAL RESEARCH

Comparison of Cape Town and Skirrow's Campylobacter isolation protocols in humans and broilers in Morogoro, Tanzania Petro Jacob & Robinson H. Mdegela & Hezron Emmanuel Nonga

Accepted: 3 February 2011 / Published online: 26 February 2011 # Springer Science+Business Media B.V. 2011

Abstract Comparison of Cape Town and Skirrow's protocols used in isolation of Campylobacter in humans and broilers was carried out in a cross-sectional study in Morogoro, Tanzania. A total of 176 and 158 human stool and broiler intestinal samples were collected, respectively. While human stool samples were collected from selected health centers, broiler intestinal samples were obtained from selected farms and chicken markets. Samples were inoculated and cultured in duplicate using two protocols and prevalence of Campylobacter were established. In humans, the prevalence of Campylobacter isolates was significantly higher (P0.05) higher Campylobacter isolation rate (54.4%) was recorded in broilers with 8 weeks compared to 7 (14.6%) and 9 weeks (8.9%).

Campylobacter isolation in humans Discussion The prevalence of 9.1% (n=16) and 21.6% (n=38) were recorded with Skirrow's and Cape Town protocols, respectively. The proportion of isolates was significantly higher (P< 0.001) with isolation using Cape Town protocol than the Skirrow's protocol. C. jejuni was the predominant species isolated with Cape Town protocol which accounted to 92.1% (n=35) of all the isolates. C. coli and C. lari were isolated at 5.3% (n=2) and 2.6% (n=1), respectively. Similarly, in Skirrow's protocol, C. jejuni was isolated at 87.5% (n=14), while C. coli and C. lari were at 6.3% (n=1) and 6.3% (n=1), respectively (Table 1). The isolation of Campylobacter spp. from human samples with respect to age strata, sex, and stool consistency is summarized in Table 2. For age, isolation of 25.5% (n=13) and 20% (n=25) were recorded in stratum I and stratum II, respectively, and the difference was not significant (P>0.05). While in females, the prevalence of Campylobacter species was 22.1% (n=19), in males, it was 21.2% (n=19), but the difference was insignificant (P>

This study has demonstrated a significant high prevalence of thermophilic Campylobacter in both humans and broilers chickens with Cape Town protocol when compared to Skirrow's protocol. To the best of our knowledge, this is the first study to use Cape Town protocol in Tanzania. Several other studies have been conducted with Skirrow's protocol which reported prevalence of Campylobacter in humans at the range of 9.3% to 18% (Lindblom et al. 1995; Mdegela et al. 2006). In broilers, the prevalence of 50% to 69% has been recorded (Mdegela et al. 2006; Chuma 2008; Kusiluka et al. 2009). The prevalence of Campylobacter reported in these previous studies are in line with the results of the current study with Skirrow's protocol. However, with Cape Town protocol, the results of this study were in agreement with work published by Le Roux and Lastovica (1998); Engberg et al. (2000); Lastovica and Le Roux (2001) and Lastovica (2006), which suggested that Cape Town protocol was superior over Skirrow's protocol on Campylo-

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Table 1 Comparison of Skirrow's and Cape Town protocols in isolation of Campylobacter from human and broiler samples Samples

Isolation protocols Sample (n) Number of OR isolates (%)

Broiler cecal content Cape Town Skirrow's Human stools Cape Town Skirrow's

158 158 176 176

123 105 38 16

(77.8) (66.5) (21.6) (9.1)

95% CI for OR P value Number of Campylobacter isolated (%)

1.77

1.23–2.57

0.03

2.75

1.42–5.52

0.001

C. jejuni

C. coli

C. lari

112 98 35 14

9 6 2 1

2 2 1 1

(91.1) (92.5) (92.1) (87.5)

(7.3) (5.7) (5.3) (6.3)

(1.6) (1.9) (2.6) (6.3)

OR odds ratio, 95%CI for OR 95% confidence limit for odds ratio, P value the χ2 test's probability value

bacter isolation from human and animal samples. Furthermore, Cape Town protocol has been considered to be simple, efficient, and cost-effective alternative method for Campylobacter isolation in humans and animals (Le Roux and Lastovica 1998; Lastovica and Le Roux 2001). It is therefore recommended that the protocol may be used in various health amenities including medical laboratories, health centers, dispensaries, and hospitals in Tanzania for Campylobacter isolation. In the present study, Cape Town protocol improved human stool cultures positive from 9.1% to 21.6%. In broiler chickens, the prevalence was improved from 66.5% to 77.8%. Such higher Campylobacter isolation has not been reported in Tanzania. This is supported by other studies conducted elsewhere, in which Cape Town protocol improved stool culture positive from 7.1% recorded earlier with Skirrow's protocol to 21.8% (Le Roux and Lastovica 1998). This shows that the magnitude of infection previously reported in humans and broilers was underestimated. The higher prevalence recorded in humans suggest that the magnitude of Campylobacter infection is big and posses a great threat as an emerging public health problem in Tanzania. Chickens being the main carrier of the pathogens were also found to have high prevalence which may be the likely major source of human campylobacteriosis. Studies that have been carried out with Cape Town protocol yielded more Campylobacter spp. compared to Skirrow's protocol (Le Roux and Lastovica 1998; Lastovica and Le Roux 2001; Lastovica 2006). In this study, three

different Campylobacter spp. including C. jejuni, C. coli, and C. lari were identified. This narrower species diversity identified during this study might have been due to inadequate identification technique, as only limited biochemical tests were used. The difference in species isolations may also be attributed by common Campylobacter spp. circulating in the local environment, sampling techniques employed, seasonality, and laboratory methodologies used. Based on this ground, further studies on speciation of the Campylobacter isolates with Cape Town protocol in Tanzania using more biochemical tests and advanced Campylobacter identification techniques are recommended. In the current study, C. jejuni constituted the majority of the identified Campylobacter spp. in humans and broilers in both Cape Town and Skirrow's protocols. This agrees with the findings of other Campylobacter studies (Kassa et al. 2005; Mdegela et al. 2006; Chuma 2008; Kusiluka et al. 2009) which observed significantly higher identification of C. jejuni than C. coli in humans and broilers. The predominance of C. jejuni in humans and chickens further suggests the involvement of this species in Campylobacterassociated enteritis in man. Indeed, C. jejuni may be the common species that may be circulating from broilers to humans and vice versa in Morogoro and possibly in Tanzania. Contrary from many other earlier findings that reported higher Campylobacter infections in young (children) than adult individuals (Ali et al. 2003; Rani et al. 2004; Mdegela et al. 2006; Chuma 2008), the results of the current study

Table 2 Isolation of Campylobacter spp. from human samples with respect to age strata, sex, and stool consistency Factor

Category

Age

Stratum I15 years Female Male Diarrhea Semisolid Solid

Sex Stool consistency

Sample (n)

Isolates (n)

Prevalence (%)

C. jejuni (%)

C. coli (%)

C. lari (%)

OR

95% CI

P value

51 125 86 90 38 110 28

13 25 19 19 11 23 4

25.5 20.0 22.1 21.2 28.9 20.9 14.4

92.3 92.0 94.7 89.5 90.9 95.7 75.0

0.0 8.0 5.3 5.3 0.0 4.3 25.0

7.7 0.0 0.0 5.3 9.1 0.0 0.0

1.37

0.6–3.1

0.55

1.06

0.5–2.3

0.98

1.00 0.65 0.41

0.15

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showed that there was no significant difference in prevalence of Campylobacter in humans with respect to age strata. Although Campylobacter infections in humans has been reported to decline with age (Coker et al. 2002), a comprehensive community-based cohort study in Egypt showed that infection could be pathogenic regardless the age of an individual, underscoring the need for strengthening prevention and control strategies for campylobacteriosis in all age groups (Rao et al. 2001). Furthermore, the calamity of HIV/AIDS which is currently increasing at an alarming rate especially in sub-Saharan Africa is worthy considered. It has been stipulated that because of HIV/AIDS, the number of campylobacteriosis in adult population in sub-Saharan Africa has rapidly increased (Coker et al. 2002). Previously published work by Mdegela et al. (2006) reported that there were no significant difference of prevalence of Campylobacter between male and female individuals which is in line with our findings. This suggests that both males and females are equally predisposed to source of Campylobacter infection. However, Coker and Adefeso (1994), Ali et al. (2003), and Chuma (2008)) recorded significantly higher Campylobacter isolation in males than females which are yet to be substantiated (Chuma 2008). In broilers, the most susceptible age recorded with Campylobacter infection (84.3%) was 8 weeks. However, the decline of prevalence from 84.3% to 66.7% was observed in chickens of 9 weeks of age in which the reason was not established. Previous studies in Tanzania also reported that Campylobacter isolation increase as the chicken age increases (Mdegela et al. 2006; Chuma 2008). Chicks can become colonized with Campylobacter within 1 to 7 days of hatching, and the bacterial burden may reach up to 1.2×107 CFU/g as the age increases (Bull et al. 2006; Horrocks et al. 2009). Based on the findings of this study, majority of broilers in Morogoro were slaughtered at the age of 8 weeks which suggests farmers, chicken meat processor, and consumers may be at higher great risk of being infected with Campylobacter where hygienic practices are not adhered to. In conclusion, the present study has introduced Cape Town protocol which is thought to be superior and efficient in Campylobacter isolation from human and animal samples in Tanzania. The protocol can be used for diagnosis of campylobacteriosis both in humans and animals and where possible to replace the commonly used Skirrow's protocol. Furthermore, broilers have been found to have high prevalence of Campylobacter infection. Farmers, farm workers, workers at broiler slaughter centers, processors at all levels, and consumers need to adopt standard hygienic practices of handling and processing of broiler meat so as to minimize the contamination of meat and meat processing facilities with Campylobacter which in

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turn may pose risk to the public. Awareness of the necessity for hand washing after contact with broilers and their products and the importance of proper cooking and handling of foods of poultry origin are probably as important in preventing Campylobacter infection. Generally, public health education programs on personal and environmental hygiene are strongly recommended to reduce potential Campylobacter public health risks in humans. Acknowledgements The authors extend much thanks to Belgian Technical Cooperation (BTC) for funding this work. Laboratory technicians in the Department of Veterinary Medicine and Public Health are thanked for their technical assistance. Patients, owners, and workers of selected Health facilities, broiler farms, restaurants, and chicken markets are gratefully acknowledged for their good cooperation during sample collection.

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