Microflora of the Dygestive System of Freshwater Turtles Trachemys sp. near the Center of Zagreb

Gastrointestinalna mikroflora gmazova sastoji se od aerobnih i anaerobnih, gram-negativnih i gram-pozitivnih bakterija, gljivica i protozoa. Većina izolata opisani su kao oportunistički patogeni koji mogu izazvati bolest u imunokompromitiranih ljudi i životinja. Podaci o bolestima koje kornjače roda Trachemys mogu proširiti na druge populacije kornjača nisu poznate, no poznato je da mogu biti rezervoar bakterija roda Salmonella te su zbog toga prijetnja zdravlju ljudi, osobito imunokompromitiranih pojedinaca. Osim salmonela, poluvodene kornjače mogu u svom probavnom sustavu nositi i brojne druge oportunističke bakterije, koje pod određenim okolnostima mogu uzrokovati bolest u ljudi i životinja. Svrha ovog istraživanja bila je steći detaljniji uvid u mikrofloru probavnog sustava kornjača roda Trachemys, te dobiti što više podataka o njezinom sastavu i eventualnim mikroorganizmima koji mogu naštetiti zdravlju ljudi i autohtonih populacija kornjača. U tu svrhu,obrisci usne šupljine i klokake od ukupno 30 kornjača uzeti su nakon kliničkog pregleda i manuelne fiksacije životinja. Uzorkovane kornjače, ulovljene su u bari nedaleko centra grada Zagreba, na području Zoološkog vrta. Obrisci su obrađeni u mikrobiološkom laboratoriju Zavoda za bolesti peradi s klinikom Veterinarskog fakulteta Sveučilišta u Zagrebu. Obrisci su nasađeni na neutralni agar i krutu selektivnu hranjivu podlogu, te potom inkubirani 24 sata pri sobnoj temperaturi. Bakterije su identificirane bojanjem po Gramu, testom tvorbe katalaze, citokrom oksidaze i API biokemijskim testovima. Za dokaz gljivica uzorci su nasađeni na Sabouraudov agar i inkubirani pet dana na sobnoj temperaturi. Iz obrisaka usne šupljine kornjača izolirano je ukupno 15 različitih vrsta bakterija, od čega je devet vrsta gram-negativnih (Proteus vulgaris, Alcaligenes, faecalis, Enterobacter cloacae, Morganella morganii, Aeromonas hydrophila, Serratia fonticola, Escherichia coli, Chryseobacterium indologenes, Brevundimonas vesicularis) i šest vrsta gram-pozitivnih bakterija (Bacillus sp., Micrococcus spp., Micrococcus luteus, Staphylococcus sp., Staphylococcus xylosus, Staphylococcus sciuri). Iz obrisaka kloake kornjača izolirano je ukupno 19 različitih vrsta bakterija, od kojih je 11 gram-negativnih (Proteus vulgaris, Enterobacter cloacae, Alcaligenes faecalis, Aeromonas hydrophila, Chryseobacterium indologenes, Aeromonas salmonicida, Morganella morganii, Enterobacter gergoviae, Kluyvera ascorbata i Citrobacter freundii) i osam gram-pozitivnih bakterija (Bacillus sp., Micrococcus luteus, Staphylococcus sp. Streptococcus sp., Bacillus cereus, Micrococcus sp., Staphylococcus xylosus i Staphylococcus sciuri). Iz obrisaka usne šupljine i kloake identificirane su gljivice Mucor sp., Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Cladosporium sp., Penicillium sp., Candida albicans i Rhodotorula rubra. Većina izoliranih bakterija i gljivica predstavljaju normalnu mikrofloru staništa poluvodenih kornjača i njihovog probavnog sustava, no ipak mogu biti i potencijalni patogeni u životinja koje su ozlijeđene ili pod stresom, te imunokompromitiranih ljudi. Informacije o mikroflori probavnog sustava ove vrste kornjača uvelike mogu rasvjetliti i njihovu invazivnost, te utjecaj na autohtone populacije kornjača.Gastrointestinal microflora of reptiles consists of aerobic and anaerobic, gram-negative and gram-positive bacteria, fungi and protozoa. Most isolates are described as opportunistic pathogens that can cause disease in immunocompromised humans and animals. Data on the diseases of the Trachemys sp. that can be extended to other turtle populations is not known but it is known that they can be reservoars of Salmonella and are therefore a threat to the health of humans, particular immunocompromised individuals. Apart from Salmonella turtles also carry numerous other opportunistic bacteria in their digestive system, which under certain circumstances can cause a disease in humans and animals. The purpose of this study was to gain a more detailed insight into the microbial digestive tract system of Trachemys sp. and to obtain as much information as possible about its composition and possible microorganisms that could harm the health of humans and native turtle populations. For this purpose, the swabs of mouth and cloaca were taken from 30 turtles after physical examination and manual fixation. Turtles are caught in a pond near the center of Zagreb, in the Zoo area. Samples were analyzed in the laboratory of the Department of Avian Diseases with Clinic on Faculty of Veterinary Medicine, University of Zagreb. Swabs were plated on Nutrient agar and Brilliant green agar and incubated for 24 hours at room temperature. The bacteria were identified by Gram staining, catalase and cytochrome oxidase tests and API biochemical tests. Samples for fungi identifications were plated on Sabouraud agar and incubated for five days at room temperature. From the swabs taken from oral cavity we isolated 15 different types of bacteria from which were nine gram-negative (Proteus vulgaris, Alcaligenes, faecalis, Enterobacter cloacae, Morganella morganii, Aeromonas hydrophila, Serratia fonticola, Escherichia coli, Chryseobacterium indologenes and Brevundimonas vesicularis) and six gram-positive bacterias (Bacillus sp., Micrococcus spp., Micrococcus luteus, Staphylococcus sp., Staphylococcus xylosus and Staphylococcus sciuri). From the swabs taken from cloaca we isolated 19 different types of bacteria from which 11 were gram-negative (Proteus vulgaris, Enterobacter cloacae, Alcaligenes faecalis, Aeromonas hydrophila, Chryseobacterium indologenes, Aeromonas salmonicida, Morganella morganii, Enterobacter gergoviae, Kluyvera ascorbata and Citrobacter freundii) and eight were gram-positive bacterias (Bacillus sp., Micrococcus luteus, Staphylococcus sp. Streptococcus sp., Bacillus cereus, Micrococcus spp., Staphylococcus xylosus and Staphylococcus sciuri). Most of the isolated bacteria and fungi represent normal flora of the environment and the gastrointestinal system of fresh water turtles, but may be potential pathogens in animals that are injured or stressed and immunocompromised humans. Information on the microflora of the digestive system of this type of turtle can greatly clarify their invasiveness as well as their impact on native turtle population