Internatinal Jurnal f Pultry Science 3 (2): 129-134, 2004 Asian Netwrk fr Scientific Infrmatin 2004 Epidemilgical Study n the Clnizatin f Chickens with Campylbacter in Briler Farms in Malaysia : Pssible Risk and Management Factrs Saleha A.A. Faculty f Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangr, Malaysia Abstract: Nineteen chicken flcks frm ten briler farms were studied fr the clnizatin f Campylbacter. A variety f factrs such as farm lcatin and chicken huse structure, water surce, rearing practice and hygiene management were investigated. Each flck was sampled weekly, frm day-ld-chicks t slaughterage chickens and envirnmental samples which include water, feed, wd shavings, flies and chicken huse envirnment were cllected t examine fr the presence f Campylbacter. In all farms, Campylbacter was nt detected in ne- and seven-day-ld chicks. Campylbacter was first detected in 38.2% f 14-day-ld-chicks and 45.3% f 21-day-ld chickens. Samples f feed, wd shavings, flies and chicken huse envirnment were all negative while nly 1.5% f untreated water supplies were fund psitive fr campylbacters. Prevalence f campylbacters clnizatin was pssibly assciated with untreated water, presence f ther animals and unhygienic management practices; als flying birds culd be a surce as they were fund t harbur campylbacters; in ne farm where 'fishing net' was placed ver the chicken huse t prevent birds frm entering, Campylbacter was nt islated in the chickens up t slaughter age. Key wrds: Campylbacter, clnizatin, briler chicken, envirnmental factrs Intrductin Campylbacterisis is reprted as ne f the mst cmmn bacterial gastrenteritis in humans, ften caused by cnsumptin r handling f pultry meat r pultry prducts. Several studies have shwn that the gastrintestinal tracts f pultry, namely chickens are frequently clnized by Campylbacter and that the prevalence f Campylbacter jejuni in briler chickens may be as high as 90.0% (Genigergis et al., 1986; Jnes at al., 1991). Since these campylbacters can survive rutine slaughtering and dressing prcessing peratins, a high prprtin f chicken carcasses sld at retail utlets are fund t be cntaminated and that up t 25.0% f freshly laid eggs may be cntaminated (Kazwala, 1993). The surce f C. jejuni in briler chickens at prductin level is nt clear althugh a number f studies have been carried ut (Genigergis et al., 1986; van de Griessen et al., 1992; Humprey et al., 1993; Kazwala et al., 1993). Frm these and ther studies, several factrs were suspected t be the surce r vectr f campylbacters in chickens; amng these factrs were the envirnment f the pultry huses, farm persnnel, presence f animals such as dgs, cats and pests as well as insects and farm hygiene and management practices. This study was undertaken t investigate the surce(s) f campylbacters in clnizing the briler chickens in the farms. Materials and Methds The farms: Ten briler farms were studied, with farms raising abut 5000 t 25 000 birds. The briler huses ranged frm ne t six per farm, depending n the size f the farm with at least 2500 t 5000 birds per huse. The briler huses were f pen type with natural ventilatin, with rfs made f either crrugated zinc sheets r nipah palm leaves (as thatch rfs) and the chickens were raised either n deep litter system r raised flr system with slatted flr. The farms were either lcated in palm il estates r n clear land. A number f the farms used tap water while thers used pnd water which were pumped directly int the water tanks situated at the rf tp f the briler huses. Islatin f campylbacters frm chicks and chickens: The large farm were represented by tw t three flcks and the smallhlder farms (with 5000 briler chickens) by ne flck. The farms received day-ldchicks frm a number f hatcheries. At each farm, clacal swabs were taken frm abut 25 t 30 day-ldchicks per flck upn their arrival. The chicks and chickens were sampled every week at each farm till they reached slaughtering age, abut 6-7 weeks ld. Islatin f campylbacters frm feed, water and the envirnment: At least 50g f feed were taken frm each farm - frm newly pened bag as well as thse in the feed trays. As fr the water, samples f at least 100 ml each, were taken frm the water tanks and frm the drinkers. Envirnmental samples cnsisted f swabs mistened with sterile peptne water which were used t swab the 129
Table 1: The descriptin f the farms and the chicken huses that were studied Parameter Range / Type N. f farms per type N. f chickens per farm 5000-22000 5000-10000 = 5 farms 10000-22000 = 5 farms N. f huses per farm 1-6 1-3 = 5 farms 4-6 = 5 farms Rearing (grwing ) days 42-45 42 days = 7 farms 45 days = 3 farms Husing system Deep litter r raised n Deep litter = 5 farms slatted flr system Raised n slatted flr = 5 farms Empty days 14-30 days 14 days = 6 farms 21-30 days = 4 farms Presence f ther animals Yes r n Yes = 6 farms N = 4 farms Surce f water supply Surface water (pnd), Surface water = 6 farms tap water (pipe) Pipe = 4 farms Lcatin On clear land r in il Clear land = 7 farms palm estate Palm il estate = 3 farms Rf type Zinc sheets r thatch rf Zinc sheets = 4 farms Thatch rf = 6 farms walls and the flrs f the briler huses; als wd T islate and estimate the number f Campylbacter shavings, dust and flies were included. These samples in water samples, the mst prbable number (MPN) were taken frm empty huses nly, that is, a week methd, described by Bltn et al. (1982) with sme befre day-ld-chicks were brught in. Wd shavings mdificatins by Humphrey and Muscat (1989), was were used in brders fr day-ld-chicks until they were used. One (1) ml prtin f each water sample was abut tw weeks ld. pipetted int 10 biju bttles, each cntaining 5 ml f All samples were taken aseptically and each placed in enriched Brucella brth (see abve). The bttles were separate sterile plastic bags r bttles. then incubated at 37 C fr 48 h. After incubatin, the brth was streaked nt Campylbacter Selective Bld Assessment f farm management and hygiene - Free Agar (Oxid) cntaining cefperasne [32 mg/l] practices: The management and hygiene practices at and amphtericin B [10 mg/l]. The plates were incubated the farms were bserved as well infrmatin gathered micr aerbically at 42 C fr 48 h. thrugh a set f questinnaires frwarded t the farm The identificatin f suspected clnies f supervisrs and/r wrkers. Campylbacter was dne as described previusly Amng the questins cvered were husbandry and (Saleha, 2002). The plates streaked with brth hygiene practices during the rearing perid, cleaning cntaining water samples were read fr the presence f and disinfectin practices and time perid when the Campylbacter and the number f campylbacter - huses were empty; cntact with animals, including negative brth were recrded. The MPN f presence f pests such as rats and flying birds. campylbacters per ml present in each water sample was read frm a table derived frm the frmula f Islatin and identificatin f Campylbacter: Clacal Campbell ( 1974 ). swabs and envirnmental samples as well as feed and water samples were brught t the labratry fr culture. Statistical analysis: Statistical tests f significance was All samples were examined fr the presence f used t assess the significance f differences in Campylbacter. The culture technique and identificatin islatin rates between grups. prcedures fr clacal swabs were as described previusly (Saleha, 2002). The feed, wd shavings and huseflies (Musca dmestica) caught in the briler huses, were each Results Briler farms and huses: Ten briler farms were studied. See Table 1. placed in separate enrichment brth, made up f Brucella brth (BBL) supplemented with cefperasne [32 mg/l], amphtericin B [10 mg/l], sdium pyruvate [0.25 g/l], sdium metabisulphite [0.25 g/l] and ferrus sulphate [0.25 g/l]. The bttles were then incubated at 37 C fr 24 h in a micr aerbic atmsphere. 130 Prevalence f campylbacters in day-ld chicks t slaughter-age chickens: Each f the 10 farms was sampled weekly (ne visit/week/farm) frm day-ldchicks till they reached slaughter age. The number f chicks and chickens sampled per visit per farm varied
Table 2: Risk factrs investigated fr pssible campylbacters clnizatin in briler chickens at the farms Pssible factrs N. f farms N. f flcks N. f Campylbacter - psitive flcks (%) (n=10) (n=19) Flck size <5000 5 6 5 (83.3) >5000 5 13 13 (100.0) Husing type deep litter 5 11 11 (100.0) raised flr 5 8 7 (87.5) Surce f water supply pipe/tap 4 5 4 (80.0) pnd/surface 4 14 14 (100.0) Lcatin clear land 7 7 6 (85.5) il palm estate 3 12 12 (100.0) Presence f ther animals Yes 6 13 13 (100.0) N 4 6 5 (83.3) Type f rf zinc sheets 4 8 8 (100.0) thatch 6 11 10 (90.9) frm 25 t 90, depending n the size f the farm. In all assciatin with the clnizatin f campylbacters in farms, campylbacter was nt islated in 1-day-ld (455 briler chickens. These factrs were flck size, husing chicks) and 7-day-ld chicks (465 chicks). The detectin type, surce f water supply, lcatin f the farm, f the initial presence f campylbacters in the farms presence f ther animals n the farm and type f rf was as fllws: in 6 farms campylbacters were as shwn in Table 2. The results shwed that the six islated in 38.2% (191 f 500) 14-day-ld chicks; in 3 factrs were nt assciated with risk f clnizatin (p farms campylbacters were islated in 45.3% (77 f value fr all six factrs > 0.1). 170) 21-day-ld chickens while in ne farm, campylbacter was nt islated in the chicks and chickens, even at slaughter age. Occurrence f Campylbacter in feed, water and envirnmental samples Feed: Campylbacter was nt islated frm 113 samples f feed frm newly pened bags as well as in the feed trays. Water: A ttal f 206 water samples were taken frm water tanks in the chicken huses and drinkers. Three (1.5%) samples were psitive fr campylbacters - samples frm water tanks in three different farms using untreated water frm the pnds. Wd shavings: All 34 samples were negative fr Campylbacter. Flies and huse envirnment: Campylbacter was nt islated frm the 30 flies and frm any 114 samples frm the chicken huses envirnment which cnsisted f tw swab samples each f the walls, flrs and dust frm a ttal f 19 chicken huses. Pssible risk factrs fr campylbacters clnizatin in the farms: Six factrs were analyzed fr their 131 Discussin Clnizatin refers t a benign, nn-pathlgical cmmensal relatinship that exists between a hst and an rganism r parasite. Campylbacters usually enter int this relatinship in the gastrintestinal tract f chickens (Stern, 1994). Fr an enteric rganism t establish and maintain clnizatin, it invlves a cmplex interactin between the hst and rganism and fr C. jejuni, the precise mechanism by which it clnizes is nt knwn (Stern, 1994). The surce f Campylbacter in briler chickens at prductin level is nt clear but the incidence f them present generally increases with age (Jnes et al., 1991). In this study, the surces f campylbacters in chicks in the farms were als nt clear as the factrs investigated shwed n significant assciatin with the risk f clnizatin. It has been reprted that vertical transmissin f Campylbacter infectin in pultry is highly imprbable (van de Giessen, 1992; Jnes et al., 1991). One reasn is that C. jejuni wuld nt penetrate int the cntents f the eggs (Dyle, 1984). Althugh egg penetratin studies revealed that C. jejuni culd be islated ccasinally frm the inner shell and membranes f refrigerated egg (Dyle, 1984), it des nt present a threat because C. jejuni des nt survive fr mre than
6 h in egg cntents (Neill et al., 1985). chlrinatin f water and cleaning and disinfectin f In the present study, all ne - day ld and seven - day ld shed drinking system, the prprtin f birds clnized briler chicks were fund negative fr Campylbacter with campylbacters reduced frm 81 t 7% which clnizatin. Thus, this study supprted the findings that increased t 84% when the interventin prgramme campylbacters are nt transmitted vertically frm stpped (Pearsn et al., 1993). The drinking water parents t chicks. This was because these briler samples in three farms were fund psitive fr chicks became clnized with Campylbacter at abut Campylbacter and the surces were untreated surface tw t three weeks f age. Other studies shwed similar water. findings, hwever ne study reprted clnizatin can Insects, particularly huse flies, have been shwn t ccur as early as seven days ld (Genigergis et al., play a rle in the disseminatin f Campylbacter 1986) while thers reprted at three t five weeks f age infectin in chickens (Rsef and Kapperud, 1983; Shane (Lindblm et al., 1986; Kazwala et al., 1990; Jacbs- et al., 1985). The present study failed t islate Reitsma, 1992) and at fur t eight weeks f age (Stern campylbacters frm flies; it may be because the et al., 2001). A study by Jacbs-Reitsma et al. (2001) number f flies sampled were small. The islatin f reprted n evidence fr vertical transmissin f Campylbacter frm darkling beetles and lesser Campylbacter in briler flcks in bth breeder and mealwrms inside the chicken huse by Jacbs - briler farms. Reitsma (1992) indicated a transmissin rute frm Once campylbacters have entered a flck, all the insects t briler chickens, althugh it is prbably mre chickens in the flck became clnized and stayed likely in that study the rute was frm chickens t insects clnized up t the time f slaughter (Lindblm et al., as the insects were psitive after the chickens were 1986; Kazwala et al., 1990). This finding was shwn in fund psitive. Hazeleger et al. (2001) reprted that the this study - there was an increase in the numbers f darkling beetle culd play a rle in the transmissin f psitive chickens at six r seven weeks f age Campylbacter as the micrrganisms fr a few days in cmpared t chicks at tw t three weeks ld. the beetles. In the study by Lindblm et al. (1986) n 250 brilers, Anther likely surce f C. jejuni in chicks is by the farm nne was psitive fr C.jejuni, while ther studies fund wrkers, intrducing the rganisms by their ftwear 24-72% f the flcks were Campylbacter - negative and clthing (Kazwala et al., 1990) and by tending ther (Kapperud et al., 1993; Humphrey et al., 1993). The pultry and pigs befre entering the briler huse present study fund nly ne flck r 6% negative fr (Kapperud, 1993). The wrkers had n prper wrking Campylbacter clnizatin. Amng the factrs which attire nr d they put n bts. Als, there was n may cntribute t the absence f campylbacters in this ftbath at the entrance f the huse. Mrever, hygiene particular farm were reasnably gd hygiene practices were minimal. Hence, the pssibility f crssmanagement f the farm, the wner wrked alne (n cntaminatin between huses by wrkers is high. van ther wrker) and it was the nly farm with 'fishing net' de Giessen et al. (1992) reprted that the use f placed ver the chicken huse t prevent pests and separate bts fr farm wrkers tending the flcks and birds frm entering. washing f hands befre dealing with pultry were The samples taken frm the huse envirnment in a assciated with a reduced risk f acquiring infectin with number f epidemilgical and risk factrs studies C. jejuni, while Humphrey et al. (1989) fund that dipping include litter, feeds, drinking water, insects, pests and bts in disinfectant befre farm wrkers entered briler ther animals (Kazwala et al., 1990; Humphrey et al., huse either delayed r prevented clnizatin with C. 1993; Kapperud et al., 1993). Generally, feeds and litter jejuni. The study by Crry et al. (2001) indicated that are fund t be Campylbacter - negative. Feeds are prly cleaned and disinfected transprt crates are a dried and pelleted which are likely t be unfavurable fr prbable surce f infectin with campylbacters. survival f campylbacters (Dyle and Rman, 1982). Rdents, in particular rats, and free - flying birds are The wrk n the survival f C. jejuni in briler feed fund anther likely surce which may intrduce C. jejuni t that C. jejuni underwent rapid death rate in feeds within chicks. They may be reservirs fr campylbacters. 24 h when stred at 20 C (Humphrey et al.,1993). Wd There were rats in all the farms studied as reprted by shavings are dry and resinus and their usage n the the farm wrkers and wners. The presence f rats n flr f the briler huse prbably has a bactericidal the farm has been shwn t be assciated with an effect n campylbacters as it des fr salmnellae increased risk (Kapperud et al., 1993). A study by (Olesink et al., 1971). Kasrazadeh and Genigergis (1987) fund 86.7% f rat Drinking water, in particular untreated water, was fund faeces were psitive fr C. jejuni. Rats, and als mice, t be assciated with Campylbacter clnizatin. In may cntaminate feed and water which then became the ne study, it was fund that the predminant surce f surce f C. jejuni fr the chickens. Once a few chickens C. jejuni n the farm was the water supply; when were clnized, they further cntaminated the feed and interventin prgramme was instituted, such as drinkers r water trughs and this caused the 132
clnizatin t spread. Campylbacter jejuni has been Hazeleger, W.C., G.J. Cenen and R.R. Beumer, 2001. islated frm birds, such as crws, pigens, blue magpies, sparrws and gray starlings (It et al., 1988; Kapperud and Rsef, 1983). On ccasins, during the visits t the farms, tw t three birds were seen flying in and ut f the briler huses in all the farms except the farm where n campylbacter was islated. These birds culd be the surce and spread campylbacters t the chickens. In tw separate studies n flying birds caught arund pultry farms and in crws caught arund residential areas, it was fund that 18% f the flying birds (Saleha et al., 2001) and 25.3% f the crws (Chng, 2001) harbured campylbacters. The management and hygiene practices in the farm d play imprtant rles in the clnizatin f C. jejuni in chickens (Humphrey et al., 1993; Kazwala et al., 1993). Generally, farms which practice gd management and hygiene have lwer rate f Campylbacter infectin (Kazwala et al., 1993) and may limit r prevent the hrizntal spread f C. jejuni (Humphrey et al., 1993) within the flck and between flcks. Acknwledgements The authr wuld like t thank clleagues fr their critical cmments, En. Kamarzaman Ahmad fr his technical assistance, the briler chicken farm wners fr making this study pssible and the University fr prviding financial aid. References Bltn, F.J., D.C. Hinchliffe and L. Rbertsn, 1982. A mst prbable number methd fr estimating small numbers f campylbacters in water. J. Hyg. Camb., 89: 185-190. nd Campbell, R.C., 1974. Statistics fr Bilgists. 2.ed. Lndn: Cambridge University Press. Chng, C.T., 2001. Campylbacter in crws: prevalence and antibitic resistance. Final year prject under DVM prgramme, Universiti Putra Malaysia. Crry, J.E.L., V.M. Allen, F. Aydin, H.I. Atabay, S.A. Bull, G. Dmingue and T.H. Humphrey, 2001. Clnizatin f briler chickens with thermphilic campylbacters in three different integrated pultry cmpanies during th rearing and slaughter. Abstracts f 11 Internatinal Wrkshp n Campylbacter, Helicbacter and related Organisms in Int. J. Med. Micrbil., 291 (S.31): 41. Dyle, M.P. and D.J. Rman, 1982. Sensitivity f Campylbacter jejuni t drying. J. Fd Prt., 45: 507-510. Dyle, M.P., 1984. Assciatin f Campylbacter jejuni with laying hens and eggs. Appl. Envirn. Micrbil., 47: 533-536. Genigergis, C., M. Hassuneh and P. Cllins, 1986. Campylbacter jejuni infectin n pultry farms and its effect n pultry meat cntaminatin during slaughtering. J. Fd Prt., 49: 895-903. 133 Survival f Campylbacter jejuni in darkling beetles (Alphitbius diaperinus). Abstracts f 11 th Internatinal Wrkshp n Campylbacter, Helicbacter and related Organisms in Int. J. Med. Micrbil., 291 (S.31): 37. Humphrey, T.J., A. Henley and D.G. Lanning, 1993. The clnizatin f briler chickens with Campylbacter jejuni: sme epidemilgical investigatins. Epidemil. Infect., 110: 601-607. Humphrey, T.J. and I. Muscat, 1989. Incubatin temperature and the islatin f Campylbacter jejuni frm fd, milk r water. Letts. Appl. Micrbil., 9: 137-139. It, K., Y. Kubkura, K. Kanek, Y. Ttake and M. Ogawa, 1988. Occurrence f Campylbacter jejuni in freeliving wild birds frm Japan. J. Wildlife Dis., 24: 467-470. Jacbs-Reitsma, W.F., 1992. Epidemilgy and identificatin f Campylbacter bacteria in pultry prductin. In Prceedings XIX Wrld's Pultry Cngress, Amsterdam, I: 371-374. Jacbs-Reitsma, W.F., C. Becht, T. De Vries, J. van der Plas, B. Duim and J. Wagenaar, 2001. N evidence fr vertical transmissin f Campylbacter in a study n Dutch breeder and Briler farms. Abstracts f 11 th Internatinal Wrkshp n Campylbacter, Helicbacter and related Organisms in Int. J. Med. Micrbil., 291 (S.31): 39. Jnes, F.T., R.C. Axtell, D.V. Rives, S.E. Scheideler, F.R. Tarver Jr., R.L. Wakker and M.J. Wineland, 1991. A survey f Campylbacter jejuni cntaminatin in mdern briler prductin and prcessing systems. J. Fd Prt., 54: 259-262. Kapperud, G. and O. Rsef, 1983. Avian wildlife reservir f Campylbacter fetus subsp. jejuni, Yersinia spp. and Salmnella spp. in Nrway. Appl. Envirn. Micrbil., 45: 375-380. Kapperud, G., E. Skjerve, L. Vik, K. Hauge, A. Lysaker, I. Aalmen, S.M. Ostrff and M. Ptter, 1993. Epidemilgical investigatin f risk factrs fr Campylbacter clnizatin in Nrwegian flcks. Epidemil. Infect., 111: 245-255. Kasrazadeh, M. and C. Genigergis, 1987. Origin and prevalence f Campylbacter jejuni in ducks and duck meat at the farm and prcessing plant level. J. Fd Prt., 50: 321-326. Kazwala, R.R., J.D. Cllins, R.A. Hannan and H.O'M. Crinin, 1990. Factrs respnsible fr the intrductin and the spread f Campylbacter jejuni in cmmercial pultry prductin. Vet. Rec., 121: 305-306. Kazwala, R.R., S.F.H. Jiwa and A.E. Nkya, 1993. The rle f management systems in the epidemilgy f thermphilic campylbacters amng pultry in Eastern zne f Tanzania. Epidemil. Infect., 110: 273-278.
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