Detection of Testudinid alphaherpesvirus, Chlamydia spp., Mycoplasma spp., and Salmonella spp. in free‑ranging and rescued Italian Testudo hermanni hermanni
DOI:
https://doi.org/10.12834/VetIt.1915.13833.1Keywords:
Mycoplasma agassizii, Reintroduction, Salmonella, Testudinid alphaherpesvirus, Testudo hermanni, Free‑rangingAbstract
Testudo hermanni is included as near‑threatened in the Red List of the International Union for Conservation of Nature, while T. hermanni hermanni is considered endangered in the Italian Red List. Appropriate management of smuggled or seized wild individuals is recommended before their reintroduction into the wild. Accordingly, a health monitoring study was carried out. During 2014‑2016, 133 oral swabs and 121 cloacal swabs were collected from a total of approximately 180 free‑ranging and rescued T. hermanni hermanni from eight different Italian regions to investigate the presence of DNA of Testudinid alphaherpesvirus (TeAHV), Chlamydia spp. and Mycoplasma spp. in the oral cavity, and Salmonella spp. isolates in the cloaca. Mycoplasma spp. was detected in 52 out of 87 (59.77%) of rescued and in 1 out of 46 free‑ranging (2.17%) individuals; 33 out of 53 (62.26%) Mycoplasma spp. positive samples were typed as M. agassizii by PCR. Salmonella spp. was isolated from 45 out of 121 (37.19%) cloacal swabs, typed into 14 serovars, and characterized for complete antimicrobial susceptibility. A significantly different distribution of Salmonella spp. isolates was found in 2016 in comparison with 2014 and 2015, without any difference between free‑ranging and rescued tortoises. All the tested tortoises were negative for TeAHV and Chlamydia spp. These results are considered a baseline information critical to monitor the dynamics of these microorganisms in free‑ranging and rescued populations of T. h. hermanni, and to correctly approach the management of rescued animals and possible relocation programs.
References
Archer G. A., Phillips C. A., Adamovicz L., Band M., Byrd J. & Allender M. C. 2017. Detection of copathogens in free-ranging eastern box turtles (Terrapene carolina carolina) in Illinois and Tennessee. J Zoo Wildl Med, 48, 1127–1134.
Bertelloni F., Chemaly M., Cerri D., Gall F.L. & Ebani V.V. 2016. Salmonella infection in healthy pet reptiles: Bacteriological isolation and study of some pathogenic characters. Acta Microbiol Immunol Hung, 63, 203-216.
Bertrand S., Rimhanen-Finne R., Weill F.X., Rabsch W., Thornton L., Perevoscikovs J., van Pelt W. & Heck M. 2008. Salmonella infections associated with reptiles: the current situation in Europe. EuroSurveill, 13, 18902.
Bosch S., Tauxe R.V. & Behravesh C.B. 2016. Turtle-Associated Salmonellosis, United States, 2006-2014. Emerg Infect Dis, 22, 1149-1155.
Brown D.R., Crenshaw B.C., McLaughlin G.S., Schumacher I.M., McKenna C.E., Klein P.A., Jacobson E.R. & Brown M.B. 1995. Taxonomic analysis of the tortoise mycoplasmas Mycoplasma agassizii and Mycoplasma testudinis by 16S rRNA gene sequence comparison. Int J Syst Bacteriol, 45, 348-350.
Brown M.B., McLaughlin G.S, Klein P.A., Crenshaw B.C., Schumacher I.M., Brown D.R. & Jacobson E.R. 1999. Upper respiratory tract disease in the gopher tortoise is caused by Mycoplasma agassizii. J Clin Microbiol, 37, 2262-2269.
Cannon R.M., Roe R.T. & Australian Bureau of Animal Health. 1982. Survey to measure the prevalence of a disease. In Livestock disease surveys: a field manual for veterinarians, Australian Government Publishing Service, Canberra.
Clinical and Laboratory Standards Institute (CLSI). 2016. Performance Standards for Antimicrobial Susceptibility Testing. 26th ed. CLSI supplement. Clinical and Laboratory Standards Institute, Wayne, Pennsylvania.
Corrente M., Madio A., Friedrich K.G., Greco, G., Desario C., Tagliabue S., D'Incau M., Campolo M. & Buonavoglia C. 2004. Isolation of Salmonella strains from reptile faeces and comparison of different culture media. J Appl Microbiol, 96, 709-715.
Corsalini T. 1975. Ricerche sulla frequenza delle salmonelle nelle tartarughe della provincia di Foggia (Testudo hermanni). Atti SISVET, 29, 624-627.
Corti C., Bassu L., Biaggini M., Bressi N., Capula M., Di Cerbo A.R., Di Francesco N., Di Tizio L., Fiacchini D., Lo Cascio P., Mastropasqua F., Nulchis V., Oneto F., Ottonello D., Richard J., Romano A., Satta M.G., Scillitani G., Spilinga C. & Vanni S. 2013. Aggiornamento sulla distribuzione italiana delle testuggini terrestri appartenenti al genere Testudo. In Atti II Congresso SHI Abruzzo-Molise "Testuggini e Tartarughe”, Chieti, Museo Universitario “G. d’Annunzio”, 27-29 September 2013.
Di Ianni F., Dodi P.L., Cabassi C.S., Pelizzone I., Sala A., Cavirani S., Parmigiani E., Quintavalla F. & Taddei S. 2015. Conjunctival flora of clinically normal and diseased turtles and tortoises. BMC Vet Res, 11, 91.
Dipineto L., Capasso M., Maurelli M.P., Russo T.P., Pepe P., Capone G., Fioretti A., Cringoli G. & Rinaldi L. 2012. Survey of co-infection by Salmonella and oxyurids in tortoises. BMC Vet Res, 28, 69.
Di Tizio L., Giacalone G. & Zuffi M.A.L. 2016. Testudo hermanni. In: Stoch F., Genovesi P. (ed.), Manuali per il monitoraggio di specie e habitat di interesse comunitario (Direttiva 92/43/CEE) in Italia: specie animali. ISPRA, Serie Manuali e linee guida, 141/2016, 256-257. https://www.minambiente.it/sites/default/files/archivio/allegati/rete_natura_2000/Manuale_specie_animali_2016_prima_parte.pdf accessed on 24 August 2020.
European Food Safety Authority (EFSA), European Centre for Disease Prevention and Control (ECDC). 2018. The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2016. EFSA J, 16, e05182.
European Commission (EC) 2003. Directive 2003/99/EC of The European Parliament and of the Council of 17 November 2003 on the monitoring of zoonoses and zoonotic agents, amending Council Decision 90/424/EEC and repealing Council Directive 92/117/EEC. Off J, L325, 12/12/2003.
Ebani V.V., Cerri D., Fratini F., Meille N., Valentini P. & Andreani E. 2005. Salmonella enterica isolates from faeces of domestic reptiles and a study of their antimicrobial in vitro sensitivity. Res Vet Sci, 78, 117-121.
Farkas S.L. & Gál J. 2009. Adenovirus and mycoplasma infection in an ornate box turtle (Terrapene ornata ornata) in Hungary. Vet Microbiol, 138, 169-173.
Feldman S.H., Wimsatt J., Marchang R.E., Johnson A.J., Brown W., Mitchell J.C. & Sleeman J.M. 2006. A novel mycoplasma detected in association with upper respiratory disease syndrome in free-ranging eastern box turtles (Terrapene carolina carolina) in Virginia. J Wildl Dis, 42, 279-289.
González Candela M., Martín Atance P., Seva J., Pallaré F.J. & Léon Vizcaíno L. 2005. Granulomatous hepatitis caused by Salmonella Typhimurium in a spur-thighed tortoise (Testudo graeca). Vet Rec, 157, 236-237.
Harris J.R., Neil K.P., Behravesh C.B., Sotir M.J. & Angulo F.J. 2010. Recent multistate outbreaks of human salmonella infections acquired from turtles: a continuing public health challenge. Clin Infect Dis, 50, 554-559.
Hartley M. & Lysons R. 2011. Development of the England Wildlife Health Strategy-a framework for decision makers. Vet Rec, 68, 158.
Hidalgo-Vila J., Díaz-Paniagua C., De Frutos-Escobar C., Jiménez-Martínez C. & Pérez-Santigosa N. 2007. Salmonella in free living terrestrial and aquatic turtles. Vet Microbiol, 119, 311-315.
Hotzel H., Blahak S., Diller R. & Sachse K. 2005. Evidence of infection in tortoises by Chlamydia-like organisms that are genetically distinct from known Chlamydiaceae species. Vet Res Commun, 29 (S1), 71–80.
Kabeya H., Sato S. & Maruyama S. 2015. Prevalence and characterization of Chlamydia DNA in zoo animals in Japan. Microbiol Immunol, 59, 507-515.
Kolesnik E., Obiegala A. & Marschang R.E. 2017. Detection of Mycoplasma spp., herpesviruses, topiviruses, and ferlaviruses in samples from chelonians in Europe. J Vet Diagnost Invest, 29, 820-832.
Jacobson E.R., Brown M.B., Wendland L.D., Brown D.R., Klein P.A., Christopher M.M. & Berry K.H. 2014. Mycoplasmosis and upper respiratory tract disease of tortoises: a review and update. Vet J, 201, 257-264.
Laroucau K., Ortega N., Vorimore F., Aaziz R., Mitura A., Szymanska-Czerwinska M., Cicerol M., Salinas J., Sachse K. & Caro M.R. 2020. Detection of a novel Chlamydia species in captive spur-thighed tortoises (Testudo graeca) in southeastern Spain and proposal of Candidatus Chlamydia testudinis. Syst Applied Microbiol, 43, 126071.
Lecis R., Paglietti B., Rubino S., Are B.M., Muzzeddu M., Berlinguer F., Chessa B., Pittau M. & Alberti A. 2011. Detection and characterization of Mycoplasma spp. and Salmonella spp. in free-living European tortoises (Testudo hermanni, Testudo graeca, and Testudo marginata). J Wildl Dis, 47, 717-724.
Lierz M., Hagen N., Harcourt-Brown N., Hernandez-Divers S.J., Lüschow D. & Hafez H.M. 2007. Prevalence of mycoplasmas in eggs from birds of prey using culture and a genus-specific mycoplasma polymerase chain reaction. Avian Pathol, 36, 145-150.
Marenzoni M.L., Santoni L., Felici A., Maresca C., Stefanetti V., Sforna M., Franciosini M.P., Casagrande Proietti P. & Origgi F.C. 2018. Clinical, virological and epidemiological characterization of an outbreak of Testudinid Herpesvirus 3 in a chelonian captive breeding facility: lessons learned and the first evidence of TeHV3 vertical transmission. PLos One, 13, e0197169.
Marenzoni M.L., Zicavo A., Veronesi F., Morganti G., Scuota S., Coletti M., Passamonti F., Santoni L., Natali P. & Moretta I. 2015. Microbiological and parasitological investigation on chelonians reared in Italian facilities. Vet Ital, 51, 173-178.
Marschang R.E., Gravendyck M. & Kaleta E.F. 1997. Herpesviruses in tortoises: investigations into virus isolation and the treatment of viral stomatitis in Testudo hermanni and T. graeca. Zentralbl Vet B, 44, 385-394.
Marschang R.E., Papp T., Ferretti L., Hochscheid S. & Bentivegna F. 2009. Detection and partial characterization of herpesviruses from Egyptian tortoises (Testudo kleinmanni) imported into Italy from Libya. J Zoo Wildl Med, 40, 211-213.
Martel A, Blahakm S, Vissenaekens H. & Pasmans F. 2009. Reintroduction of clinically healthy tortoises: the herpesvirus Trojan horse. J Wildlife Dis, 45, 218-220.
Mazzotti S. 2006. Testudo hermanni (pp. 386-395). In Atlante degli Anfibi e Rettili d’Italia. (Barbieri F., Doria G., & Sindaco R. eds.). Societas Herpetologica Italica. Polistampa, Firenze.
Messmer T.O., Skelton S.K., Moroney J.F., Daugharty H. & Fields B.S. 1997. Application of a nested, multiplex PCR to psittacosis outbreaks. J Clin Microbiol, 35, 2043-2046.
Ministero dell'ambiente e della tutela del territorio e del mare. 2019. Linee guida per il ricollocamento in natura di individui del genere Testudo spp. confiscati. https://www.minambiente.it/sites/default/files/archivio/allegati/cites/linee_guida_testudo_aggiornate_2019.pdf accessed on 27 August 2020.
Mitura A., Niemczuk K., Zaręba K., Zając M., Laroucau K. & Szymańska-Czerwińska M. 2017. Free-living and captive turtles and tortoises as carriers of new Chlamydia spp. PLoS One, 12, e0185407.
Origgi F.C. 2012. Testudinid Herpesviruses: A Review. J Herpetol Med Surg, 22, 42-54.
Origgi F.C. & Rigoni D. 2003. A Tortoise herpesvirus outbreak in Italy: History, Diagnosis, Therapy and Follow up. Proc. ARAV, 21-23.
Origgi F.C., Tecilla M., Pilo P., Aloisio F., Otten P., Aguilar-Bultet L., Sattler U., Roccabianca P., Romero C.H., Bloom D.C. & Jacobson E.R. 2015. A Genomic Approach to Unravel Host-Pathogen Interaction in Chelonians: The Example of Testudinid Herpesvirus 3. PLoS One, 10, 0134897.
Pasmans F., De Herdt P., Chasseur-Libotte M.L., Ballasina D.L. & Haesebrouck F. 2000. Occurrence of Salmonella in tortoises in a rescue centre in Italy. Vet Rec, 146, 256-258.
Pasmans F., De Herdt P., Dewulf J. & Haesebrouck F. 2002. Pathogenesis of infections with Salmonella enterica subsp. enterica serovar Muenchen in the turtle Trachemys scripta scripta. Vet Microbiol, 87, 315-325.
Pees M., Rabsch W., Plenz, B., Fruth A., Prager R., Simon S., Schmidt V., Munch S. & Braun P. 2013. Evidence for the transmission of Salmonella from reptiles to children in Germany, July 2010 to October 2011. Euro Surveill, 18, 20634.
Percipalle M., Giardina G., Lipari L., Piraino C., Macrì D. & Ferrantelli V. 2011. Salmonella infection in illegally imported spur-thighed tortoises (Testudo graeca). Zoonoses Public Health, 58, 262-269.
Rondinini C., Battistoni A., Peronace V. & Teofili C. 2013. Lista Rossa IUCN dei Vertebrati Italiani. Comitato Italiano IUCN e Ministero dell’Ambiente e della Tutela del Territorio e del Mare, Roma, Italia.
Rubini S., Ravaioli C., Previato S., D'Incau M., Tassinari M., Guidi E., Lupi S., Merialdi G. & Bergamini M. 2016. Prevalence of Salmonella strains in wild animals from a highly populated area of north-eastern Italy. Ann Ist Super Sanità, 52, 277-280.
Soldati G., Lu Z.H., Vaughan L., Polkinghorne A., Zimmermann D. R., Huder J. B. & Pospischil A. 2004. Detection of mycobacteria and chlamydiae in granulomatous inflammation of reptiles: a retrospective study. Vet Pathol, 41, 388-397.
Sting R., Ackermann D., Blazey B., Rabsch W. & Szabo I. 2013. Salmonella infections in reptiles--prevalence, serovar spectrum and impact on animal health. Berl Munch Tierarztl Wochenschr, 126, 202-208.
Taylor-Brown A., Rüegg S., Polkinghorne A. & Borel N. 2015. Characterisation of Chlamydia pneumoniae and other novel chlamydial infections in captive snakes. Vet Microbiol, 178, 88-93.
Thrusfield M. 2005. Surveys. In Thrusfield M. Veterinary epidemiology. 3rd edn. Blackwell Publishing, London, 228-246.
VanDevanter D.R., Warrener P., Bennett L., Schultz E.R., Coulter S., Garber R.L. & Rose T.M. 1996. Detection and analysis of diverse herpesviral species by consensus primer PCR. J Clin Microbiol, 34, 1666-1671.
Vojdani A. & Franco A.R. 1999. Multiplex PCR for the detection of mycoplasma fermentans, m. hominis, and m. penetrans in patients with chronic fatigue syndrome, fibromyalgia, rheumatoid arthritis, and Gulf War syndrome. J Chronic Fatigue Syndrome, 5, 187-197.
World Organisation for Animal Health (OIE). 2018. Salmonellosis. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.09.08_SALMONELLOSIS.pdf
