Preliminary Investigations on Salmonella spp. Incidence in Meat Chicken Farms in Italy, Germany, Lithuania and the Netherlands

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1 Internatinal Jurnal f Pultry Science 7 (8): , 2008 ISSN Asian Netwrk fr Scientific Infrmatin, 2008 Preliminary Investigatins n Salmnella spp. Incidence in Meat Chicken Farms in Italy, Germany, Lithuania and the Netherlands Juzas Pieskus, Maria Pia Francisini, Patrizia Casagrande Prietti, Felix Reich, Edvardas Kazeniauskas, Ceslva Butrimaite-Ambrzeviciene, Myklas Mauricas and Nic Blder 1 Institute f Immunlgy Vilnius University, Mletu pl. 29, L-08249, Vilnius, Lithuania 2 Dipartiment di Scienze Bipatlgiche ed Igiene delle Prduzini Animali e Alimentari, Facltà di Medicina Veterinaria, Via San Cstanz Perugia, Italy 3 Institut fur Lebensmittelqualitat und sicherheit, Tierarzliche Hchschule Hannver, Bischfhler Damm 15, D Hannver, Germany 4 Animal Sciences Grup, Wageningen UR, Divisin f Infectius Diseases, Edelhertweg 15, 8200 AB Lelystad, The Netherlands Abstract: The briler chickens, especially if intensively reared, can be cnsidered as an imprtant reservir f Salmnella infectins in humans. Many cnsumers assume that briler chickens, grwn under cnventinal cmmercial cnditins, have higher infectin levels f Salmnella than free-range rganic chickens. The subject f this study was firstly t assess the incidence f Salmnella spp. In cnventinal chicken farms, lcated in different areas f Italy, Germany, Lithuania and in the Netherlands. In additin rganic farms were investigated bth in Italy and the Netherlands. The typificatin f Salmnella was als effected in attempt t value the distributin f the rganism n the basis f the different gegraphical areas. The incidence f Salmnella in cnventinal briler farms was 29% in Lithuania, 20% in Italy and 11% in The Netherlands, while in Germany Salmnella was nt detected. Salmnella was islated frm rganic briler flcks in Italy (18, 1%) and in The Netherlands (3, 7%). Our results indicated that Salmnella Enteritidis and Salmnella Typhimurium dminated in Lithuanian briler flcks while Salmnella Infantis and Salmnella Java were predminant in the Netherlands. Salmnella hadar and Salmnella heidelberg seemed t be prevalent in Italy. Key wrds: Brilers, cnventinal, rganic, salmnella Intrductin Overbeke et al. (2006) reprted n significant differences Briler chickens, especially if intensively reared, can be in the prevalence f Salmnella between rganic and cnsidered as an imprtant reservir f Salmnella cnventinal brilers at slaughter. The subject f this infectins fr humans (EFSA, 2006). Varius risk factrs study was t assess the incidence f Salmnella spp. in can be respnsible fr Salmnella infectin and its briler farms f 4 EU cuntries: Italy, Germany, The spread in pultry farms: husing system, flck size, Netherlands and Lithuania. In Italy and in the different age f chicken and seasn f the year. (Rse et Netherlands besides the cnventinal farms, rganic al., 1999; Heyndrickx et al., 2002). Many cnsumers flcks were investigated in rder t preliminary assume that the rganic chickens shed less Salmnella determine the diffusin f Salmnella spp. in this than cnventinal brilers because f the particular rearing cnditin. The typificatin f Salmnella was system management (lw density stcking, access t als effected in attempt t evaluate the distributin f the utside and special diets) (Bailey and Csby, 2005). A rganism n the basis f the different gegraphical high number f chickens in the farm can increase the areas. chance f infectin with Salmnella, (Mllenhrst et al., 2005). On the ther hand in rganic farm the access t Materials and Methds utside may increase the risk f the infectin with Sampling: This study was perfrmed in 4 EU cuntries Salmnella thrugh the cntact with faeces f wild birds (Italy, Germany, Lithuania and the Netherlands). In Italy and ther animals. Cui et al. (2005) fund that and in the Netherlands bth rganic and cnventinal rganically raised brilers had higher prevalence f farms were investigated. In Italy the investigatins were Salmnella than brilers raised cnventinally. In perfrmed in 11 rganic flcks and in 10 cnventinal several studies the incidence f Salmnella was lwer flcks ver a perid f tw years. During the prductin in rganic than in cnventinal briler farms (Heuer et cycle envirnmental samples cnsisting f 5 pls f al., 2001; Wlf-Reuter et al., 2002). Hwever, Van litter, 2 pls f dust and 1 water sample were cllected. 813

2 Pieskus et al.: Salmnella spp. incidence in brilers At slaughterhuse 60 caecal samples per farm, subdivided in 2 pls, were cllected. In the Netherlands 18 cnventinal flck and 108 rganic flcks were examined ver a perid f tw years. Different numbers f dust, litter samples, water and caecal pl samples were cllected frm each flck (Table 1). Deliveries f rganic feed were als sampled and analysed individually. In Lithuania and in Germany the investigatins were perfrmed in 27 and 22 cnventinal flcks respectively. In Lithuania flck samples cnsisting f 5 pls f litter, 2 pls f dust and 1 water sample were cllected during the prductin cycle. Litter since it is nt heat treated samples cnsisted f bts swabs. Five (pled) samples f caecum per flck were taken frm a briler slaughterhuse. In Germany nly caecum samples were cllected and analysed as ne pled sample per flck. Dust samples were cllected frm the walls, fans and ther surfaces and pled int samples f apprximately 25 g. Per pl Farms were supplied frm lcal underwater reservir in Italia and in Lithuania, while Dutch farms were all cnnected t public water supply. In all cases the water sampled cnsisted f pl f 1 litre cllected frm nipples. Micrbilgical analysis: Salmnella were islated by standard methds (Internatinal Organizatin fr Standardizatin 6579, 1998). Frm each pl f litter, dust and caecal cntent, 10 g were hmgenised with apprximately. 1:10 Buffered peptne water (BPW) fr 60 secnds; 25 ml f each water sample were pured int a cntainer fllwed by adding 225 ml f BPW. The samples were incubated at 35±2 C fr h. And then 1±0.1 ml was transferred int 10 ml Muller Kauffmann Tetrathinate Nvbicin brth (MKTTn)brth (1.0 ml in 9.0 ml TBG at 37 C fr 24 h) (Mdified Semislid Rappaprt Vassiliadis Agar in The Netherlands) and 0.1±0.02 ml was transferred int 10 ml Rappaprt Vassiliadis Brth (RV) brth. The tubes were incubated in a water bath at 42±0.5 C fr h.the subcultures frm the enrichment media were made nt Xylse Lysine Desxychlate (XLD) agar plates and n Hecten enteric (Rambach agar in Germany and Brilliant Green Agar in the Netherlands) agar plates and then incubated aerbically at 37 C fr hurs. One lpful f inculum was used fr each plate. Presumptive clnies were inculated nt Mc Cnkey agar (Standard I agar in Germany and Brilliant Green Agar in the Netherlands), incubated at 37 C fr 24 h and bichemically checked with Ureum Agar (UA), Triple Sugar Irn Agar (TSI) using cmmercial tests (API 20 E). Identificatin was perfrmed by agglutinatin testing with agglutinatin sera. The 95% cnfidence intervals (CI) fr the bserved prevalence f Salmnella-psitive samples were estimated by linear interplatin frmula (Mntgmery Ranger, 1999): CI = p - z [p (1-p)/n], CI = p + z [p (1-p)/n] p = number f psitive samples/number f tested samples; z = (95%) 1.96; n = number f tested samples. Results All results are summarized in Table 1. In Italy 2 ut f 10 cnventinal farms were psitive fr S. hadar and S. heidelberg (20%) while 2 ut f 11 rganic flcks were psitive fr S. Hadar (18, 1%). In particular, in cnventinal farms Salmnella was islated frm 5 ut f 50 litter samples (10%; CI 1, 6-18, 3 ) and frm 4 ut f 20 caecal samples (20%; CI 2, 4-37, 5 ). In ne farm bth litter and caecal samples were Salmnella psitive, while in the ther farms it was detected nly in the intestinal samples. In the rganic farms Salmnella was islated frm 4 f 22 caecal samples,(18, 8%; CI 2, 3-33, 6). The dust and water samples were always Salmnella negative. The results btained in Lithuania shwed that the mst infected samples were litter and caecum (25, 9%; CI and 25%; CI respectively) while Salmnella presence resulted lwer in dust and water (9, 6%; CI and 3, 8%; CI , respectively). 22 cnventinal flcks, cllected ver a perid f ne year gave negative results fr Salmnella in all cases. In the Netherlands the analysis fr Salmnella was perfrmed in 11 cnventinal farms with a ttal f 18 flcks and in 108 flcks frm 16 rganic briler farms during 2 years. With respect t rganic farming a ttal f 181 litter samples were taken; frm which 4 (2, 2%) were Salmnella psitive. All samples frm dust and envirnmental samples prved t be Salmnella negative. S. infantis was islated in 1 rganic feed sample frm ne flck, while S. senftenberg was fund, bth in a feed sample and in litter samples frm anther flck. N pls f caecal samples frm rganic chickens were Salmnella psitive at slaughter. Salmnella sertypes islated frm rganic flcks were: S. infantis, S. senftenberg, ne islate frm the B grup and ne frm the C grup culd nt be typed cmpletely. In Dutch cnventinal flcks the mst infected samples resulted t be caecum (4 psitive ut f 36 = 11.1%). The sertype islates were S. infantis in the samples frm ne farm (dust, litter and caeca) and S. Java frm litter samples in ne ther farm. Discussin Varius risk factrs exist fr infectin with Salmnella spp. and its diffusin in pultry farms: husing system, flck size, different age f chicken and seasn f the year (Angen et al., 1996; Skv et al., 1999). This study was intended t investigate the incidence f Salmnella in briler chicken farms lcated in limited gegraphical areas f 4 different cuntries: Italy, Germany, Lithuania and the Netherlands. In Italy and in the Netherlands the 814

3 Pieskus et al.: Salmnella spp. incidence in brilers Table 1: Salmnella spp. prevalence in cnventinal and rganic meat chicken farms in Italy, Germany, Lithuania and The Netherlands N.f N.f N. f N. f Type f tested psitive tested psitive Cuntry prductin Sample flcks flcks samples samples CI (95%) Sertype Italy Org. Dust S. hadar, Litter Water Caecum Ttal: Cnv. Dust S. hadar, Litter Water Caecum S. heidelberg Ttal: Lithuania Cnv. Dust S. enteritidis, Litter S. typhimurium Caecum Water Ttal: The Netherlands Cnv. Env./dust S. infantis, Litter S. java Caecum Water Ttal: Org. Env./dust S. infantis Litter S. senftenberg Caecum S. Grup B Feed S. Grup C Ttal: Germany Cnv. Caecum Ttal per 4 cuntries: (10%) (3.2%) presence f Salmnella was als investigated in the cnsisted f caecal samples, always cnsidered the rganic meat type chicken in rder t evaluate a best substratum fr the Salmnella recvery. pssible influence f the rearing cnditins. Our results Salmnella islatin was als slightly inferir t indicated that the incidence f Salmnella spp. varies in expectatins in Italy. In Lithuania and in the Netherlands the cuntries investigated. The percentage f a higher prevalence fr Salmnella was detected in Salmnella islates in cnventinal husing system cmparisn with the EU Baseline study. Salmnella in was 29% in Lithuania, 11.1% in the Netherlands and 9% rganic briler flcks was bserved bth in Italy (2 ut f in Italy. In ne Dutch farm Salmnella culd be islated 11 flcks) and in The Netherlands (4 ut f 108 flcks). frm samples taken in briler huses after cleaning and The data is nt enugh extensive t make a cmparisn disinfectin (data nt shwn). Frm these data a specific between the tw rearing systems but they may give an interventin culd be started, which prved t be effective idea in relatin t the distributin f Salmnella in n the farms. The infectin was prved t persist in rganic briler farms. In the literature there is faecal remains, in cracks and jints r in the feeder cntrversial data abut the influence f husing auger system inside the briler huses. In this farm systems fr Salmnella infectin. In several studies the black beetles and larvae appeared t be a vectr fr incidence f Salmnella was lwer in rganic than in transmissin f Salmnella Java. In anther study this cnventinal briler farms (Heuer et al., 2001; Wlftype f transmissin was als fund (Blder, 2004). In Reuter et al., 2002; Van der Hulst et al., 2004). Italian the EU Baseline study n brilers (EFSA, 2007) the data indicated that the presence f Salmnella in results fr the fur cuntries invlved in this study rganic briler flcks is cmparable with cnventinal deviated frm the results f the present paper. In the briler flcks and are in agreement with thse reprted baseline study, perfrmed n the flck basis, Germany, by Van Overbeke et al. (2006), wh did nt find Italy, Lithuania and the Netherlands had respectively significant differences in the prevalence f Salmnella 15%; 28.3%; 2.9% and 7.5% f Salmnella psitive between rganic and cnventinal brilers at slaughter. flcks. Hwever the data frm ur investigatins cvered Bailey and Csby (2005) als did nt agree in assuming nly a fractin f the ttals presented in the Baseline that the free range r rganic cnditins can influence study. In Germany Salmnella in cnventinal briler the Salmnella presence in the chickens. The flcks was nt detected but it shuld be underlined that explanatin f the lwer Salmnella prevalence in Dutch the data is referred t limited sampling, thugh it rganic brilers culd be that rganic brilers grw 815

4 Pieskus et al.: Salmnella spp. incidence in brilers lnger (8 week +) than cnventinal brilers (5-6 weeks) Salmnella culd change year by year in relatin t and are mstly slw grwing brands. This culd explain different factrs, like the use f certain antibitics; fr the absence f Salmnella in caecal cntents f instance the recent distributin f S. Hadar in several previusly Salmnella psitive flcks. The prevalence areas culd be due t its resistance t flurquinlnes at the age f 8 weeks may have drpped belw the (Tran et al., 2004; Breuil et al., 2000). detectin level f the survey. Salmnella infectin experiments in cnventinal brilers shwed that even Cnclusin: The subject f this study was primary t after relatively heavy Salmnella challenge at the early assess the incidence f Salmnella spp. in meat age, brilers may have cleared the infectin at chicken farms f 4 EU cuntries: Italy, Germany, the apprximately 6 weeks f age (Blder et al., 1999; Lintn Netherlands and Lithuania. In Italy and in the et al., 1985). Feed samplings in Dutch rganic briler Netherlands rganic flcks were als investigated in farms were included since feed fr rganic brilers is rder t cmpare the Salmnella distributin in different usually nt pelletized. If it is nt heat treated it culd be rearing cnditins. Our results cannt be cnclusive, cnsidered a risk factr in transmissin f Salmnella. since they are referred t limited areas in the cuntries In this study rganic feed samples prved t be and nly Italy and the Netherlands culd prduce data Salmnella psitive and in ne ccasin the same n bth rearing systems (rganic and cnventinal), sertype was nt islated frm feed and litter samples thugh the results were nt very cmprehensive either. in the same flck. Hwever mnitring results indicate In the Netherlands and in Lithuania cnventinal that there is a negligible risk frm pelletized feed. husing system seems t cnstitute a favurable (Blder, unpublished data). Our results shwed that the cnditin fr Salmnella spread in brilers. Italian data mst infected samples were frm litter and caecum in shwed that n significant differences in the incidence cmparisn with thse f dust and water. Sme f Salmnella were fund between rganic and investigatrs have determined that the cntents f the cnventinal briler rearing system. The absence f caeca cnstitute the best single sample site fr the Salmnella in caecum samplings frm rganic chickens search f Salmnella (Barrw et al., 1988). Others have in the Netherlands can be explained by the natural cmpared sampling f litter and the use f drag swabs clearance, which cnsequently leads t the prevalence fr detectin f Salmnella in pultry flcks (Kingstn, belw detectin level f the survey with the age. 1981). Cncerning the dust samples, Salmnella was psitive in cnventinal farms in Lithuania and in the Netherlands and always negative in rganic farms. Althugh dust can be cnsidered an excellent vectr fr Salmnella, survival in dust is limited (Davies and Acknwledgements The study was part f the research prject Pultry Flrgut funded by the Eurpean Cmmissin Cntract n.: Fd-CT-200 X Breslin, 2003). Gast et al. (1998) suggested that infectin culd ccur by ral ingestin f external References surfaces cntaminated by Salmnella. Different Angen, Ø. Skv, M.N. Chriel, J.F. Agger and M. Bisgaard, Salmnella servars were identified in chicken. At EU level the mst cmmn servar was S. Enteritidis which represented apprximately half f the islates (EFSA, 2007). Our results indicated that S. Enteritidis and S. Typhimurium dminated in Lithuanian briler flcks, while in the Netherlands S. infantis and S. java were A retrspective study f Salmnella infectin in Danish briler flcks. Prev. Vet. Med., 26: Bailey, J.S. and D.E. Csby, Salmnella prevalence in free-range and certified rganic chickens. J. Fd Prtectin, 68: islated. In Italy S. hadar and S. heidelberg were fund t be prevalent. These results cnfirm thse f the EU Baseline study in brilers (EFSA, 2007) where wide variety f prevalent Salmnella servars was islated in different cuntries. Besides, ther investigatrs (Byrd et al., 1997; Ry et al., 2002) fund that S. Kentucky and S. Heidelberg were predminant sertypes islated frm pultry meat prducts. The investigatrs frm Japan (Limawngpranne et al., 1999) detected that S. blckley, S. hadar and S. bredeney were predminant in brilers, meanwhile S. enteritidis was fund nly in 0.9% f samples. It seems t prve that distributin f Salmnella servars in the wrld depends n the gegraphical regin. In additin the distributin f Barrw, P.A., J.M. Simsn and M.A. Lvell, Intestinal clnizatin in the chickens by fdpisning Salmnella sertypes; micrbial characteristics assciated with faecal extractin. Avian Pathlgy, 17: Blder, N.M., J.A. Wagenaar, F.F. Putirulan, K.T. Veldman and M. Smmer, The effect f flavphsphlipl (Flavmycin) and salinmycin sdium (Sacx) n the excretin f Clstridium perfringens, Salmnella enteritidis and Campylbacter jejuni in brilers after experimental infectin. Pult. Sci., 78: Blder, N.M., Bestrijding van Salmnella Java vereist maatwerk: De drgang hinderen. Pluimveehuderij, 34:

5 Pieskus et al.: Salmnella spp. incidence in brilers Breuil, J., A. Brisabis, I. Casin, L. Armand-Lefevre, S. Limawngpranne, S., H. Hayashidani, A.T. Okatani, K. Fremy and E. Cllatz, Antibitic resistance in On, C. Hirta, K. Kanek and M. Ogawa, Salmnellae islated frm humans and animals in Prevalence and persistence f Salmnella in briler France: cmparative data frm, 1994 and J. chickens flcks. J. Vet. Med. Sci., 61: Anti-micrbial Chemtherapy, 46: Lintn, A.H., Z.A. Al-Chalaby and M.H.Hintn,1985. Byrd, J.A., D.E. Crrier, J.R. DeLach and D.J. Nisbert, Natural subclinical salmnella infectin in chickens: Cmparisn f drag-swab envirnmental a ptential mdel fr testing the effects f varius prtcls fr the islatin f Salmnella in pultry prcedures n Salmnella shedding. Vet. Recrd, huses. Avian Dis., 41: : Cui Shengui, Beilei Ge, Jie Zheng and Jianghng Meng, Mllenhrst, H.C.J. Van Wudenberg, E.G.M. Bkkers Prevalence and Antimicrbial resistance f and I.J.M. de Ber, Risk factrs fr Campylbacter spp. and Salmnella servars in Salmnella Enteritidis infectins in Laying hens, rganic chickens frm Maryland retail stres. Appl. Pult. Sci., 84: and Envirn. Micrbi., 71: Rse, N., F. Beauden, P. Druin, J.Y. Tux, V. Rse and Davies, R.H. and M. Breslin, Persistence f P. Clin, Risk factrs fr Salmnella enterica Salmnella enteritidis type 4 in the envirnment and subsp. enterica cntaminatin in Frech briler arthrpd vectrs n an empty free range chicken chicken at the end f the rearing perid. Preventive farm. Envirn. Micrbi., 5: Vet. Med., 39: EFSA, Trends and surces f znses, zntic Ry, P., A.S. Dhilln, L.H. Lauernman, D.M. Schaberg, D. agents and antimicrbial resistance in the Bandli and J. Jhnsn, Results f Salmnella Eurpean unin in, The EFSA J., islatin frm pultry prducts, pultry, pultry EFSA, Reprt f the Task Frce n Znses envirnment and ther characteristics. Avian Dis., Data Cllectin n the Analysis f the baseline 46: survey n the prevalence f Salmnella in briler Skv, M.N., O. Angeng, M. Chrie, J.E. Olsen and M. flcks f Gallus gallus in the EU, Part A: Bisgaard, Risk factrs assciated with Prevalence estimates. The EFSA J., Salmnella enterica servar Typhimurium infectin Gast, R.K., B.W. Mitchel and P.S. Hlt, Airbrne in Danish briler flcks. Pult. Sci., 78: transmissin f Salmnella enteritidis infectin Tran, T.P., T.T. Nguyen, M. Akiba, N. Ogasawara, D. between grups f chicken in cntrlled- Shinda, A.T. Okatani and H. Hayashidani, envirnmental islatin cabinets. Avian Dis., 42: Prevalence f Salmnella spp. in pigs, chickens and ducks in the Mekng Delta, Vietnam. Heuer, O.E., K. Pedersen, J.S. Andersen and M. Madsen, Van der Hulst-van M.C. Arkel, R.P. Kwakkel, T.B Prevalence and antimicrbial susceptibility f Rdenburg, Salmnella and Campylbacter thermphilic Campylbacter in rganic and nd in rganic briler prductin system. In 22 Wrld s cnventinal briler flcks. Letter in App. Micrbi., Pultry Cngress, Istanbul, Turkey, p: : Van Overbeke, I., L. Duchateau, L. De Zutter, G. Albers Heyndrickx, M., D. Vandekerchve, L. Herman, I. Rllier, and R. Ducatelle, A cmparisn survey f K. Grijspeerdt and I. De Zutter, Rutes fr rganic and cnventinal briler chickens fr Salmnella cntaminatin f pultry meat: infectius agents affecting health and fd safety. epidemilgical study frm hatchery t Avian Dis., 50: slaugterhuse. Epidemilgy Infectin, 129: 253- Wlf-Reuter, M., S. Matthes and F. Ellendrf, Salmnella prevalence in intensive, free and Kingstn, D.J., A cmparisn f culturing drag rganic prductin systems. Archiv für swabs and litter fr identificatin f infectins with Gefügelkunde, 66: 158. Salmnella spp. in cmmercial chickens flcks. Avian Dis., 25:

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