Investigating public support for biosecurity measures to mitigate pathogen transmission through the herpetological trade.

The expanding global trade in herpetofauna has contributed to new infectious disease dynamics and pathways that allow for the rapid spread of pathogens geographically. Improved biosecurity is needed to mitigate adverse biodiversity, economic and human health impacts associated with pathogen transmission through the herpetological trade. However, general lack of knowledge of the pathogen transmission risks associated with the global trade in herpetofauna and public opposition to biosecurity measures are critical obstacles to successfully preventing pathogen transmission. In 2019 we administered a survey to 2,007 members of the public in the United States of America to ascertain their support for interventions to prevent the spread of Batrachochytrium dendrobatidis (Bd), Batrachochytrium salamandrivorans (Bsal), ranaviruses, and Salmonella through the herpetological trade. We presented survey respondents with different potential hazards associated with pathogen transmission through this trade, namely ecological, economic, and human health impacts. We used structural equation models to determine how these different hazards and respondents' characteristics influenced respondents' support for quarantine and veterinary observation of herpetofauna imported into the United States, mandatory tests for diseases of concern, and best practices to reduce stress and improve the care of live herpetofauna during transport to the United States. Respondents' values and their perceived susceptibility and sensitivity to different hazards associated with pathogen transmission were key determinants of their support for biosecurity. Respondents with strong biospheric and altruistic values demonstrated sensitivity to ecological and human health impacts associated with pathogen transmission, whereas respondents with strong egoistic values demonstrated sensitivity to economic impacts. Respondents had limited knowledge of Bd, Bsal or ranaviruses, the size of the herpetological trade, or how this trade may contribute to pathogen transmission. Improved outreach and education on pathogen transmission through the herpetological trade is required, but it is important that messages are tailored to people with different values to elicit their support for biosecurity.

Vertebrate host phylogeny influences gut archaeal diversity.

Commonly used 16S rRNA gene primers do not detect the full range of archaeal diversity present in the vertebrate gut. As a result, several questions regarding the archaeal component of the gut microbiota remain, including which Archaea are host-associated, the specificities of such associations and the major factors influencing archaeal diversity. Using 16S rRNA gene amplicon sequencing with primers that specifically target Archaea, we obtained sufficient sequence data from 185 gastrointestinal samples collected from 110 vertebrate species that span five taxonomic classes (Mammalia, Aves, Reptilia, Amphibia and Actinopterygii), of which the majority were wild. We provide evidence for previously undescribed Archaea-host associations, including Bathyarchaeia and Methanothermobacter, the latter of which was prevalent among Aves and relatively abundant in species with higher body temperatures, although this association could not be decoupled from host phylogeny. Host phylogeny explained archaeal diversity more strongly than diet, while specific taxa were associated with both factors, and cophylogeny was significant and strongest for mammalian herbivores. Methanobacteria was the only class predicted to be present in the last common ancestors of mammals and all host species. Further analysis indicated that Archaea-Bacteria interactions have a limited effect on archaeal diversity. These findings expand our current understanding of Archaea-vertebrate associations.

Molecular detection of Wolbachia endosymbiont in reptiles and their ectoparasites.

Wolbachia, a maternally transmitted Gram-negative endosymbiont of onchocercid nematodes and arthropods, has a role in the biology of their host; thus it has been exploited for the filariasis treatment in humans. To assess the presence and prevalence of this endosymbiont in reptiles and their ectoparasites, blood and tail tissue as well as ticks and mites collected from them were molecularly screened for Wolbachia DNA using two sets of primers targeting partial 16S rRNA and Wolbachia surface protein (wsp) genes. Positive samples were screened for the partial 12S rRNA and cytochrome c oxidase subunit 1 (cox1) genes for filarioids. Of the different species of lizards (Podarcis siculus, Podarcis muralis and Lacerta bilineata) and snakes (Elaphe quatuorlineata and Boa constrictor constrictor) screened from three collection sites, only P. siculus scored positive for Wolbachia 16S rRNA. Among ectoparasites collected from reptiles (Ixodes ricinus ticks and Neotrombicula autumnalis, Ophionyssus sauracum and Ophionyssus natricis mites), I. ricinus (n = 4; 2.8%; 95% CI, 0.9-7) from P. siculus, N. autumnalis (n = 2 each; 2.8%; 95% CI, 0.9-6.5) from P. siculus and P. muralis and O. natricis (n = 1; 14.3%; 95% CI, 0.7-55.4) from Boa constrictor constrictor scored positive for Wolbachia DNA. None of the positive Wolbachia samples scored positive for filarioids. This represents the first report of Wolbachia in reptilian hosts and their ectoparasites, which follows a single identification in the intestinal cells of a filarioid associated with a gecko. This data could contribute to better understand the reptile filarioid-Wolbachia association and to unveil the evolutionary pattern of Wolbachia in its filarial host.

Role of reptiles and associated arthropods in the epidemiology of rickettsioses: A one health paradigm.

We assessed the presence of Rickettsia spp., Coxiella burnetii and Anaplasma phagocytophilum in reptiles, their ectoparasites and in questing ticks collected in a nature preserve park in southern Italy, as well as in a peri-urban area in another region. We also investigated the exposure to these pathogens in forestry workers, farmers and livestock breeders living or working in the nature preserve park given the report of anecdotal cases of spotted fever rickettsioses. Rickettsia spp. were molecularly detected in Podarcis muralis and Podarcis siculus lizards (i.e., 3.1%), in Ixodes ricinus (up to 87.5%) and in Neotrombicula autumnalis (up to 8.3%) collected from them as well as in I. ricinus collected from the environment (up to 28.4%). Rickettsia monacensis was the most prevalent species followed by Rickettsia helvetica. An undescribed member of the family Anaplasmataceae was detected in 2.4% and 0.8% of the reptiles and ectoparasites, respectively. Sera from human subjects (n = 50) were serologically screened and antibodies to Rickettsia spp. (n = 4; 8%), C. burnetti (n = 8; 16%) and A. phagocytophilum (n = 11; 22%) were detected. Two ticks collected from two forestry workers were positive for spotted fever group (SFG) rickettsiae. Ixodes ricinus is involved in the transmission of SFG rickettsiae (R. monacensis and R. helvetica) in southern Europe and lizards could play a role in the sylvatic cycle of R. monacensis, as amplifying hosts. Meanwhile, N. autumnalis could be involved in the enzootic cycle of some SFG rickettsiae among these animals. People living or working in the southern Italian nature preserve park investigated are exposed to SFG rickettsiae, C. burnetii and A. phagocytophilum.

The Phylogenetic Structure of Reptile, Avian and Uropathogenic Escherichia coli with Particular Reference to Extraintestinal Pathotypes.

The impact of the Gram-negative bacterium Escherichia coli (E. coli) on the microbiomic and pathogenic phenomena occurring in humans and other warm-blooded animals is relatively well-recognized. At the same time, there are scant data concerning the role of E. coli strains in the health and disease of cold-blooded animals. It is presently known that reptiles are common asymptomatic carriers of another human pathogen, Salmonella, which, when transferred to humans, may cause a disease referred to as reptile-associated salmonellosis (RAS). We therefore hypothesized that reptiles may also be carriers of specific E. coli strains (reptilian Escherichia coli, RepEC) which may differ in their genetic composition from the human uropathogenic strain (UPEC) and avian pathogenic E. coli (APEC). Therefore, we isolated RepECs (n = 24) from reptile feces and compared isolated strains' pathogenic potentials and phylogenic relations with the aforementioned UPEC (n = 24) and APEC (n = 24) strains. To this end, we conducted an array of molecular analyses, including determination of the phylogenetic groups of E. coli, virulence genotyping, Pulsed-Field Gel Electrophoresis-Restriction Analysis (RA-PFGE) and genetic population structure analysis using Multi-Locus Sequence Typing (MLST). The majority of the tested RepEC strains belonged to nonpathogenic phylogroups, with an important exception of one strain, which belonged to the pathogenic group B2, typical of extraintestinal pathogenic E. coli. This strain was part of the globally disseminated ST131 lineage. Unlike RepEC strains and in line with previous studies, a high percentage of UPEC strains belonged to the phylogroup B2, and the percentage distribution of phylogroups among the tested APEC strains was relatively homogenous, with most coming from the following nonpathogenic groups: C, A and B1. The RA-PFGE displayed a high genetic diversity among all the tested E. coli groups. In the case of RepEC strains, the frequency of occurrence of virulence genes (VGs) was lower than in the UPEC and APEC strains. The presented study is one of the first attempting to compare the phylogenetic structures of E. coli populations isolated from three groups of vertebrates: reptiles, birds and mammals (humans).

Bacteriophage as an alternative to prevent reptile-associated Salmonella transmission.

Salmonellosis is a major global public health issue; its most common infection, gastroenteritis, accounts for approximately 90 million illnesses and 150,000 mortalities per year. Eradicating salmonellosis requires surveillance, prevention and treatment, entailing large expenditures. However, it is difficult to control Salmonella transmission because it occurs via multiple routes; exotic reptiles are a reservoir of Salmonella and comprise one such route. As the popularity of exotic pets and animal exhibition has increased, human encounters with reptiles have also increased. As a result, reptile-associated salmonellosis (RAS) has been recognized as an emerging disease. The development of antimicrobial resistance in RAS-causing Salmonella sp. requires alternatives to antibiotics. In this study, bacteriophages have been established as an alternative to antibiotics because only target bacteria are lysed; thus, they are promising biocontrol agents. Here, bacteriophage pSal-SNUABM-02, which infects and lyses reptile Salmonella isolates, was isolated and characterized. The morphology, host range, growth traits and stability of the phage were investigated. The phage was assigned to Myoviridae and was stable in the following conditions: pH 5-9, 4-37°C, and ultravioletA/ultravioletB (UVA/UVB) exposure. Salmonella clearance efficacy was tested using planktonic cell lysis activity and biofilm degradation on polystyrene 96-well plates and reptile skin fragments. The phage exhibited vigorous lysis activity against planktonic cells. In in vitro biofilm degradation tests on reptile skin and polystyrene plates, both low- and high-concentration phage treatments lowered bacterial cell viability by approximately 2.5-3 log colony-forming units and also decreased biomass. Thus, bacteriophages are a promising alternative to antibiotics for the prevention and eradication of RAS.

Molecular detection of the reptile-associated Borrelia group in Amblyomma dissimile, Mexico.

Recently, the first record of Borrelia associated with reptiles in Mexico was published; however, no studies have been done to assess the role of Mexican ticks as potential vectors of this Borrelia group. Amblyomma dissimile is a hard tick mainly associated with amphibians and reptiles in this country. The aim of this study was to evaluate the presence of Borrelia in A. dissimile from Mexico. We collected 60 A. dissimile individuals attached to 16 Rhinella horribilis. DNA was extracted and all specimens were screened individually for Borrelia by amplification of a fragment of the 16S rDNA and an additional fragment of the flagellin gene. Five ticks were positive for Borrelia, DNA sequences corresponded to Borrelia sp. and group with sequences of the reptile-associated Borrelia group. This is the first report of Borrelia in A. dissimile and the second report of the reptile-associated Borrelia group in Mexico. This study also highlights the importance of this tick species as potential vector of this group.

Molecular detection of vector-borne agents in ectoparasites and reptiles from Brazil.

Trombidiformes and Mesostigmata mites, as well as Ixodida ticks, infest ectothermic tetrapods worldwide, potentially acting as vectors of bacteria, viruses and protozoa. The relationship among ectoparasites, transmitted pathogenic agents (e.g., Borrelia spp., Coxiella spp., Hepatozoon spp., and Rickettsia spp.) and ectothermic hosts has been scarcely investigated. This research focuses on a large collection of Brazilian herpetofauna screened for the presence of arthropod ectoparasites and vector-borne microbial agents. Reptiles (n = 121) and amphibians (n = 49) from various locations were infested by ectoparasites. Following genomic extraction, microbial agents were detected in 81 % of the Acari (i.e. n = 113 mites and n = 26 ticks). None of the mites, ticks and tissues from amphibians yielded positive results for any of the screened agents. Blood was collected from reptiles and processed through blood cytology and molecular analyses (n = 48). Of those, six snakes (12.5 %) showed intraerythrocytic alterations compatible with Hepatozoon spp. gamonts and Iridovirus inclusions. Hepatozoon spp. similar to Hepatozoon ayorgbor and Hepatozoon musa were molecularly identified from seven hosts, two mite and two tick species. Rickettsia spp. (e.g., Rickettsia amblyommatis, Rickettsia bellii-like, Rickettsia sp.) were detected molecularly from four mite species and Amblyomma rotundatum ticks. Phylogenetic analyses confirmed the molecular identification of the above-mentioned microbial agents of mites and ticks related to snakes and lizards. Overall, our findings highlighted that the Brazilian herpetofauna and its ectoparasites harbour potentially pathogenic agents, particularly from the northern and south-eastern regions. The detection of several species of spotted fever group Rickettsia pointed out the potential role of ectothermic hosts and related arthropod ectoparasites in the epidemiological cycle of these bacteria in Brazil.

Global Prevalence of Chlamydial Infections in Reptiles: A Systematic Review and Meta-Analysis.

Background and Objectives:Chlamydia spp. are potential zoonotic pathogens that can infect a wide range of animal hosts. In reptiles, Chlamydia can cause hepatitis, pneumonitis, and conjunctivitis and it can cause high mortality in young animals. The objectives of this study were to estimate the pooled prevalence of chlamydial infections in reptiles and to assess the trend of these infections over time. Materials and Methods: The study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Relevant studies were retrieved from PubMed, Scopus, and Web of Science. The retrieved studies were screened for eligibility. Then, important data were extracted from the included studies. A random effects model was used for all analyses. Subgroup analysis was used to assess heterogeneity for orders of reptiles, continents where the studies were conducted, and types of specimens. Cumulative meta-analysis and meta-regression were used to determine the trend of the prevalence over time. The quality of each included study was evaluated. Results: Of 106 studies (with a total of 2607 samples), 20 met the inclusion criteria and were included in the meta-analysis. The pooled prevalence of chlamydial infections in reptiles was 23.5% (95% confidence interval [CI]: 15.4-34.0). The trend of chlamydial infections increased from 1990 to 2008; thereafter, it was almost stable at slightly over 20%. The most commonly reported Chlamydia spp. were Chlamydia psittaci, Chlamydia pneumoniae, Chlamydia pecorum, and Chlamydia caviae. Among reptiles, the prevalence of chlamydial infections was highest in crocodiles (57.3% [95% CI: 32.5-78.9]). Among continents, the prevalence of chlamydial infections was highest in Australia (68.6% [95% CI: 36.8-89.1]). Conclusions: Based on the included studies, the prevalence of chlamydial infections in reptiles was high, especially in crocodiles. Because C. psittaci and C. pneumoniae are commonly found in reptiles and are well-known zoonotic pathogens, they should be of concern for human health.

Challenges in microbiological identification of aerobic bacteria isolated from the skin of reptiles.

INTRODUCTION: Bacterial pathogens are often involved in dermatitis in reptiles. Exact identification of reptile-specific but otherwise uncommon bacterial species may be challenging. However, identification is crucial to evaluate the importance of the detected bacterial species. OBJECTIVE: The aim of this study was to assess the number of aerobic bacterial isolates cultured from skin-derived samples of reptiles which were not reliably identified by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS), and to determine their identity. MATERIAL AND METHODS: Routine bacterial diagnostics were performed on 235 skin samples, and 417 bacterial isolates were analysed by MALDI-TOF MS. The isolates were grouped into categories based on their first score: category I (≥ 2.00), category II (≥ 1.70 and < 2.00), and category III (< 1.70). Isolates from category III were further investigated by 16S rRNA gene sequencing and the following criteria were applied: query cover 100%, e-value rounded to 0.0 and sequence identity (%) > 98.00% for genus identification, and > 99.00% for species identification. RESULTS: The majority of bacterial isolates were in category I (85.1%) or category II (8.4%). In category III (6.5%) results achieved at first by MALDI-TOF MS corresponded to the results of the molecular analysis in 8.0% of isolates at the species level and in 24.0% at the genus level. Bacterial isolates classified as category III were heterogenic in genus (e.g. Chryseobacterium, Devriesea, Pseudomonas, Staphylococcus, Uruburuella), and some have only been described in reptiles so far. CONCLUSIONS: Most of the aerobic bacterial isolates cultured from reptile skin achieved high scores by MALDI-TOF MS. However, in the majority of category III isolates MALDI-TOF MS results were different from those of the molecular analysis. This strengthens the need to carefully examine low-scored results for plausibility and to be familiar with the occurrence and morphology of relevant reptile-specific bacterial species (e.g. Devriesea agamarum) as well as with the limits of the database used.

[Aerobic bacteria from skin lesions in reptiles and their antimicrobial susceptibility]. / Aerobes Keimspektrum und Resistenzlage bei Hautläsionen von Reptilien.

OBJECTIVE: Bacterial skin infections are common in reptiles. Although many such infections are influenced by multifactorial problems, specific treatment of bacterial infections is an important consideration. The objective of this study was to evaluate the range of aerobic bacteria in skin lesions of reptiles and to determine their antimicrobial susceptibility. MATERIAL AND METHODS: Swabs of skin lesions from 219 reptiles were cultured for aerobic bacteria between January 2017 and June 2018. Isolates were identified based on growth on selective agar plates, biochemical parameters, as well as MALDI-TOF MS. Antibiotic susceptibility testing was carried out using the microdilution method. RESULTS: A total of 306 isolates were identified, mostly gram-negative, including Pseudomonas spp. (n = 48), Citrobacter spp. (n = 31, only in chelonians), aerobic spore-forming bacteria (n = 30), Aeromonas spp. (n = 20), Acinetobacter spp. (n = 20), Proteus spp. (n = 15), Staphylococcus spp. (n = 15), Klebsiella spp. (n = 13), Enterococcus spp. (n = 13), Morganella spp. (n = 11) as well as 78 other gram-negative and 12 other gram-positive bacteria. Colonization with 2 (n = 80) or more (n = 16) bacterial isolates was seen in 96 animals. Antibiotic susceptibility testing was carried out with 208 of the 306 isolated bacteria. Many isolates were sensitive (minimal inhibitory concentration [MIC] in µg/ml ≤ breakpoint) to enro- (E) and marbofloxacin (M): 86.4 % MIC ≤ 0.5 (E) and 95.5 % MIC ≤ 1 (M) for Pseudomonas spp., 86.4 % MIC ≤ 0.5 (E) and 90.9 % MIC ≤ 1 (M) for Citrobacter spp., 75.0 % MIC ≤ 0.5 (E) and 100 % MIC ≤ 1 (M) for Aeromonas spp. Trimethoprim/sulfamethoxazol proved to be effective against most of the Citrobacter spp. (90.9 % MIC ≤ 2/38) and Aeromonas spp. (75.0 % MIC ≤ 2/38). Amikacin was effective against nearly all Pseudomonas spp. (97.7 % MIC ≤ 16), Citrobacter spp. (95.5 % MIC ≤ 16) and Aeromonas spp. (93.8 % MIC ≤ 16). CONCLUSION AND CLINICAL RELEVANCE: The majority of isolates were gram-negative; the clinical relevance of individual isolates must, however, be evaluated on a case by case basis. Many of the isolated bacteria were sensitive to fluoroquinolones as well as aminoglycosides. Susceptibility testing is recommended since use of these antibiotics should be limited and for every tested group of antibiotics resistant isolates were found.

Emerging Infectious Diseases of Chelonians: An Update.

Chelonians are increasingly challenged by anthropogenic threats and disease. This article summarizes recent literature and clinical experiences regarding 4 emerging infectious diseases in turtles and tortoises: ranaviruses, cryptosporidiosis, intranuclear coccodiosis of Testudines, and Emydomyces testavorans.

Prevalence of Salmonella serovars isolated from reptiles in Norwegian zoos.

BACKGROUND: Reptiles are known to be asymptomatic carriers of Salmonella spp. in their gastrointestinal mucosa and a variety of Salmonella serovars including exotic serovars mainly associated with reptiles as well as human pathogenic serovars have been isolated. There are many case reports of reptile-associated Salmonella infections worldwide, including one case in Norway in 2000. In August 2017, there was a legislative change in Norway that allowed more permissive reptile ownership and legalized the keeping of 19 different reptile species by private persons. There has been a concern that this new legislation will lead to an increase in reptile-associated salmonellosis in Norway, however knowledge is lacking on the occurrence of Salmonella spp. in Norwegian reptiles. The aim of this study was therefore to investigate the prevalence of Salmonella spp. in captive reptile species in Norway, identify the serovars and evaluate their zoonotic potential. Thus, cloacal swabs were taken from 53 snakes, 15 lizards and 35 chelonians from three Norwegian zoos, and assessed for the presence of Salmonella spp. by culture, biochemical testing and serotyping. RESULTS: In total, 43% of the reptiles were shedding Salmonella spp., with a prevalence of 62%, 67% and 3% in snakes, lizards and chelonians, respectively. A total of 26 different serovars were found, including Salmonella enterica spp. enterica (40%) and S. enterica spp. arizonae (4%), both of which are considered to have a high zoonotic potential. S. enterica spp. diarizonae, salamae and houtenae were also identified, however these serovars are considered to have a lower zoonotic potential. CONCLUSIONS: The current study demonstrates that captive Norwegian reptiles are carriers of potentially zoonotic Salmonella spp. Given the increasing popularity of reptiles as pets and the legislative change, reptile-associated salmonellosis could become an increasingly important public health concern in Norway. Adequate public information about the risk of Salmonella infection as well as preventive measures to avoid Salmonella transmission from reptiles to humans is needed. The risk of Salmonella infection is considered low when recommended precautions are taken and good hygiene exhibited.

Comparison of detection methods for Salmonella enterica shedding among reptilian patients at a veterinary teaching hospital.

In the United States, ~1.4 million sporadic human Salmonella enterica infections occur annually, with an estimated 6% attributable to reptile exposure. Detection of Salmonella in reptiles can be challenging given the limitations among detection methods. We evaluated sampling and detection methods for S. enterica in a cross-sectional study of reptilian patients (n = 45) over the course of 13 mo. Two sampling methods (cloacal swabs, electrostatic cloth body-feet samples) and 3 detection methods (enriched culture, lateral flow immunoassay [LFI], real-time PCR) were compared using McNemar and Fisher exact tests. Results varied by species, sample type, and detection method. In total, 14 of 45 (33%) patients were positive by culture, 10 of 45 (22%), and/or 13 of 45 (29%) by rtPCR. Among rtPCR-positive results, cloacal swabs (12 of 45 [27%]) resulted in a higher detection than body-feet wipes (4 of 45 [9%]; p = 0.01). Among culture-positive results, shedding was most commonly detected after additional incubation at room temperature when testing cloacal swabs (9 of 45 [20%]). However, there was significant disagreement between sampling methods (cloacal vs. body-feet; p = 0.03). No samples were positive by LFI. In general, cloacal swabs yielded the highest test-positive rates, irrespective of testing method. Our study highlights the importance of using detection methods optimized for the sample being tested.

Source Attribution of Salmonella in Macadamia Nuts to Animal and Environmental Reservoirs in Queensland, Australia.

Salmonella enterica is a common contaminant of macadamia nut kernels in the subtropical state of Queensland (QLD), Australia. We hypothesized that nonhuman sources in the plantation environment contaminate macadamia nuts. We applied a modified Hald source attribution model to attribute Salmonella serovars and phage types detected on macadamia nuts from 1998 to 2017 to specific animal and environmental sources. Potential sources were represented by Salmonella types isolated from avian, companion animal, biosolids-soil-compost, equine, porcine, poultry, reptile, ruminant, and wildlife samples by the QLD Health reference laboratory. Two attribution models were applied: model 1 merged data across 1998-2017, whereas model 2 pooled data into 5-year time intervals. Model 1 attributed 47% (credible interval, CrI: 33.6-60.8) of all Salmonella detections on macadamia nuts to biosolids-soil-compost. Wildlife and companion animals were found to be the second and third most important contamination sources, respectively. Results from model 2 showed that the importance of the different sources varied between the different time periods; for example, Salmonella contamination from biosolids-soil-compost varied from 4.4% (CrI: 0.2-11.7) in 1998-2002 to 19.3% (CrI: 4.6-39.4) in 2003-2007, and the proportion attributed to poultry varied from 4.8% (CrI: 1-11) in 2008-2012 to 24% (CrI: 11.3-40.7) in 2013-2017. Findings suggest that macadamia nuts were contaminated by direct transmission from animals with access to the plantations (e.g., wildlife and companion animals) or from indirect transmission from animal reservoirs through biosolids-soil-compost. The findings from this study can be used to guide environmental and wildlife sampling and analysis to further investigate routes of Salmonella contamination of macadamia nuts and propose control options to reduce potential risk of human salmonellosis.

Mycoses are emerging in reptiles / Emergen las micosis en reptiles

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Comparison of the phylogenetic analysis of PFGE profiles and the characteristic of virulence genes in clinical and reptile associated Salmonella strains.

BACKGROUND: Salmonella is generally considered as a human pathogen causing typhoid fever and gastrointestinal infections called salmonellosis, with S. Enteritidis and S. Typhimurium strains as the main causative agents. Salmonella enterica strains have a wide host array including humans, birds, pigs, horses, dogs, cats, reptiles, amphibians and insects. Up to 90% of reptiles are the carriers of one or more serovars of Salmonella. Extraintestinal bacterial infections associated with reptiles pose serious health threat to humans. The import of exotic species of reptiles as pet animals to Europe correlates with the emergence of Salmonella serotypes, which not found previously in European countries. The presented study is a new report about Salmonella serotypes associated with exotic reptiles in Poland. The goal of this research was to examine the zoonotic potential of Salmonella strains isolated from reptiles by comparative analysis with S. Enteritidis strains occurring in human population and causing salmonellosis. RESULTS: The main findings of our work show that exotic reptiles are asymptomatic carriers of Salmonella serovars other than correlated with salmonellosis in humans (S. Enteritidis, S. Typhimurium). Among the isolated Salmonella strains we identified serovars that have not been reported earlier in Poland, for example belonging to subspecies diarizonae and salamae. Restriction analysis with Pulsed-field Gel Electrophoresis (PFGE), showed a great diversity among Salmonella strains isolated from reptiles. Almost all tested strains had distinct restriction patterns. While S. Enteritidis strains were quite homogeneous in term of phylogenetic relations. Most of the tested VGs were common for the two tested groups of Salmonella strains. CONCLUSIONS: The obtained results show that Salmonella strains isolated from reptiles share most of virulence genes with the S. Enteritidis strains and exhibit a greater phylogenetic diversity than the tested S. Enteritidis population.

Identification and Characterization of Escherichia coli, Salmonella Spp., Clostridium perfringens, and C. difficile Isolates from Reptiles in Brazil.

Considering the increasing popularity of reptiles as pets and their possible role as reservoirs of pathogenic microorganisms, the aim of this study was to isolate Escherichia coli, Salmonella spp., Clostridium perfringens, and C. difficile strains from reptiles in Brazil and to characterize the isolated strains. The characterization was based on phylogenetic typing of E. coli, identification of virulence genes of E. coli, C. perfringens, and C. difficile, serotyping of Salmonella spp., ribotyping and MLST of C. difficile and antimicrobial susceptibility test of pathogenic strains. Cloacal swabs were collected from 76 reptiles, of which 15 were lizards, 16 chelonians, and 45 snakes, either living in captivity, in the wild, or as companion animals. E. coli was isolated from 52 (68.4%) reptiles, of which 46 (88.4%) were characterized as phylogroup B1. The virulence factor CNF1 of E. coli was found in seven (9.2%) sampled animals, whereas the gene of EAST1 was found in isolates from two (2.6%) reptiles. Three isolates positive for CNF1 were resistant to cephalothin, one of which was also resistant to ciprofloxacin, trimethoprim/sulfamethoxazole, and chloramphenicol, being then classified as multidrug resistant strain (MDR). Salmonella enterica was identified in 26 (34.2%) reptiles, of which 13 belonged to the subspecies enterica. Serotypes such as S. Mbandaka, S. Panama, S. Infantis, S. Heidelberg, and S. Anatum were identified. One isolate of S. enterica subsp. houtenae was resistant to cephalothin and ciprofloxacin. C. perfringens type A was isolated from six (7.8%) animals. C. difficile was isolated from three (3.9%) reptiles. Two of these isolates were toxigenic and classified into ribotypes/MLST 081/ST9 and 106/ST42, which have been previously reported to infect humans. In conclusion, reptiles in Brazil can harbor toxigenic C. difficile and potentially pathogenic E. coli and Salmonella enterica subsp. enterica, thus representing a risk to human and animal health.