Albamonte Rodríguez, Andreu

Universitat Autònoma de Barcelona. Facultat de Veterinària

Diposit Digital de Documents de la UAB

THE ROLE OF WILD ANIMALS AS A RESERVOIR OFZOONOTIC AGENTS : Clostridium difficileAndreu Albamonte Rodríguez Bachelor’s Degree Final Thesis Veterinary Medicine School, UAB                    June 2017 Accipiter nisus A total of 235 samples were analysed by microbiologyand 123 by PCR. No direct C. difficile isolation was obtained in theselective media culture. Only 1 out of the 123 PCR-analysed samples waspositive for the toxin A gene (tcdA). The sample camefrom an Eurasian Sparrowhawk (Accipiter nisus). Noneof the samples were positive for the toxin B gene(tcdB).4.        RESULTSPCR  positive sample for tcdA by PCRAccipitriformes (13%)Apodiformes (3%)Bucerotiformes(2%)Anseriformes (2%)Passeriformes(13%)Strigiformes(18 %)Others (3%)Columbiformes(3%)Charadriiformes(13%)Mammals(n=93)Birds(n=118)Reptiles (n=11)Others (5%)Genetta genetta(3%)Martes foina (2%)Caproeluscaproelus(3%)Cervuselaphus(3%)Erinaceuseuropaeus(43%)Sus scrofa(27%)Mustela vison (14%)Testudo horsfieldii (9%)Mauremysleprosa (18%)Testudohermanni(18%)TrachemysScripta(46%)Falconiformes (20%)3.1  Sampling representation4.1  Prevalence of Clostridium difficile4.2 Other potential zoonotic bacteriaPure cultures isolated from wildlife (n=51) Antibiotic Resistance (AR) Testing for Escherichia coli (n=12)3.2 Laboratorial sampling process5.   DISCUSSION1.   INTRODUCTIONSensitiveIntermediateResistantThe highest prevalence source of C difficile infections havebeen described in pig and cattle farms. There are few studiesreporting CD toxigenic strains in farm pest species such asrodents and shrews (Jardine et al. 2013; Himsworth et al.2014; Andrés-Lasheras et al. 2017). However, studies fromfarming-free areas or Nature Parks such as Doñana in Spain(Alderete et al. 2011) or in Slovenia (Bandelj et al. 2011)obtained zero prevalence of CDI in wild animals and in wildpasserine birds respectively.In our study, a cloacal swab from an Accipiter nisus waspositive for the toxin A gene of CD by PCR. This is the firstdescription of a CDI positive wild raptor in Catalonia. Thisanimal came from the Maresme (Catalonia) and it washospitalized due to a mild electrocution. The Accipiter nisusis one of the smaller size raptors in Spain, which habits innon-urbanized, wooded areas, catching small birds.Catalonia is a region with a high density of porcine farms.Although we cannot determine the origin of this infection, itmight be supposed that the Accipiter nisus could get infectedby CD predating small birds and rodents that were in closecontact to pig farms . As regards the microbiological results,the fact that no CD isolation could be seen even after theethanol treatment for activating the spore germination, couldbe explained by the low burden load of wildlife as reservoirsof CDI. So, it is important to complement the detection usinga much more sensitive method like the PCR.Finally, the prevalence of multidrug resistant (MDR) bacteriasuch as E.coli or Klebsiella pneumoniae in wild animals, wasanother relevant fact. Most of E. coli isolates (75%) wereresistant to 3 or more antimicrobials, as previously reported(Pinto et al. 2015; Vidal et al. 2016). The highest percentageof resistance were to Ampicillin (91,6%), Cephalexin (83,3%)and Amoxicillin-Clavulanic acid (66,6%). Moreover, K.pneumoniae was only sensitive to 11% antibiotics(Chloramphenicol and Tobramycin). All samples of the studyprovided from non-treated wild animals suggesting thesource of MRD bacteria infection could be the pollution of theenvironment by the high density of livestock farms and theintensity of antimicrobials treatments. To assess the prevalence of toxigenic strains of C. difficile (tcdA and tcdB) in wildlife of Catalonia. To determine the presence of other potential zoonotic enteropathogens in wildlife of Catalonia.Gram stain of C. difficileAR Testing Klebsiellapneumoniae (n=1)Wildlife attended at the Wildlife Rehabilitation Center (WRC) of Torreferrusa (Catalonia, Spain) were sampled at the first moment of the admission during the period betwen October of 2016and May of 2017 Fecal or cloacal samples were systematically analysed by means of standarized microbiological and molecular techniques as shown in the scheme (3.2)Laddersamples LadderC+ C+Selective culture of C. difficile6. CONCLUSIONS• This is the first description of a wild raptor positiveby PCR to C difficile toxigenic A strain in Catalonia.• The presence of MDR bacteria in no-drug-treatedwildlife is a real fact.• The source of these infections could be theenvironment pollution secondary to intensivelivestock production.• The presence of these zoonotic bacteria in wildanimals should be considered of relevance at theanimal-human-ecosystem interface since they candisseminate them through their migratory routes.The ecology and epidemiology of wildlife diseases, particularly those that are zoonotic, must become an area of focus for public health programs(Siembieda et al., 2011). Many studies have demonstrated that wild animals are reservoirs of zoonotic pathogens that can represent a risk for thehuman health, such as viral zoonotic pathogens (West Nile or Influenza A), parasites (Toxoplasma gondii) and multidrug-resistant zoonotic bacteria(Vidal et al., 2016). However, there are few studies in wildlife related to emergent pathogens.Clostridium difficile (CD) is considered one of the most important nosocomial agent producing severe disease (diarrhea, colitis, toxic megacolon) andmortality in hospitalized patients. This pathogen is a Gram-positive, anaerobic, spore-forming bacillus, with the capacity of producing toxins that areresponsible of these human deaths. In the last years, CD has also been associated with outcomes of diarrhea in the general population (communitycases) suggesting the involvement of livestock (pig, beef, poultry) as a reservoirs of CD infection (CDI). However, the role of wild animals in theepidemiology of CDI is scarcely investigated. For this reason, the main objective of this study has been to determine the presence of wildlife asreservoirs of CD toxigenic strains2.     OBJECTIVES 3.    MATERIAL and METHODS Alderete PA, Eulalia M, Sánchez G, Luis J, Cancelo B. 2011. Universidad Complutense de Madrid Facultad de Veterinaria Departamento de Sanidad Animal; Clostridium difficile;Prevalencia e  importancia ecológica en animales domesticos y fauna salvaje. Bandelj P, Trilar T, Racnik J, Zadravec M, Pirš T, Avbersek J, Micunovic J, Ocepek M, Vengust M. 2011. Zero prevalence of Clostridium difficile in wild passerine birds in Europe. FEMS Microbiol. Lett. 321:183–185. Himsworth CG, Patrick DM, Mak S, Jardine CM, Tang P, Weese JS. 2014. Carriage of Clostridium difficile by wild urban Norway rats (Rattus norvegicus) and black rats (Rattus rattus). Appl. Environ. Microbiol. 80:1299–305. Jardine CM, Reid-Smith RJ, Rousseau J, Weese JS. 2013. Detection of Clostridium difficile in small and medium-sized wild Mammals in Southern Ontario, Canada. J. Wildl. Dis. 49:418–21. Pinto, A., Simoes, R., Oliveira, M. Vaz-Pires, P., Brandao, R., da Costa, P.M. 2015. Multidrug resistance in wild bird populaions: importance of the food chain. J Zoo Wildl Med, 46, 723-31.Vidal, A., Baldomà, L., Molina-Lopez. R., Martin. M., Darwich. L., 2017. Microbiological diagnosis and antimicrobial sensitivity profiles in diseased free-living raptors. J Med Mic. 61, 837-43.7. Bibliography

The role of wild animals as a reservoir of zoonotic agents : clostridium difficile

http://ddd.uab.cat/pub/tfg/2017/179831/TFG_aalbamonterodriguez_poster.pdf

The role of wild animals as a reservoir of zoonotic agents : clostridium difficile

Abstract

Similar works

Full text

Available Versions

Diposit Digital de Documents de la UAB