Isolation of Brucella inopinata from a White's tree frog (Litoria caerulea): pose exotic frogs a potential risk to human health?

Introduction: Cold-blooded hosts, particularly exotic frogs, have become a newly recognized reservoir for atypical Brucella species and strains worldwide, but their pathogenicity to humans remains largely unknown. Here we report the isolation and molecular characterization of a B. inopinata strain (FO700662) cultured from clinical samples taken from a captive diseased White's Tree Frog (Litoria caerulea) in Switzerland. The isolation of B. inopinata from a frog along with other reports of human infection by atypical Brucella raises the question of whether atypical Brucella could pose a risk to human health and deserves further attention. Methods: The investigations included histopathological analysis of the frog, bacterial culture and in-depth molecular characterization of strain FO700662 based on genome sequencing data. Results and Discussion: Originally identified as Ochrobactrum based on its rapid growth and biochemical profile, strain FO700622 was positive for the Brucella- specific markers bcsp31 and IS711. It showed the specific banding pattern of B. inopinata in conventional Bruce-ladder multiplex PCR and also had identical 16S rRNA and recA gene sequences as B. inopinata. Subsequent genome sequencing followed by core genome-based MLST (cgMLST) analysis using 2704 targets (74% of the total chromosome) revealed only 173 allelic differences compared to the type strain of B. inopinata BO1T, while previously considered the closest related strain BO2 differed in 2046 alleles. The overall average nucleotide identity (ANI) between the type strain BO1T and FO700622 was 99,89%, confirming that both strains were almost identical. In silico MLST-21 and MLVA-16 also identified strain FO700662 as B. inopinata. The nucleotide and amino acid-based phylogenetic reconstruction and comparative genome analysis again placed the isolate together with B. inopinata with 100% support. In conclusion, our data unequivocally classified strain FO700622, isolated from an exotic frog, as belonging to B. inopinata.

DETECTION OF AN ARENAVIRUS IN A GROUP OF CAPTIVE WAGLER'S PIT VIPERS (TROPIDOLAEMUS WAGLERI).

A group of eight Wagler's pit vipers (Tropidolaemus wagleri) from a private collection died with respiratory signs within 6 mo of one another. The group consisted of an adult breeding pair that was wild caught and six offspring from this pair. Four of the dead snakes were submitted for gross and histopathology. Signs of bacterial pneumonia were detected in all four examined snakes. No inclusion bodies suggestive of viral infection were found in any of the examined tissues. Polymerase chain reactions for the detection of ferla-, adeno-, reo-, and nidoviruses were all negative, but reptarenaviruses closely related to viruses previously described in boa constrictors (Boa constrictor) with inclusion body disease were detected in two of the four snakes. This is the first description of reptarenaviruses in viperid snakes. The pathogenic role of the virus in illness is unknown.

DETECTION OF OPHIDIOMYCES OPHIODIICOLA IN TWO CAPTIVE BOCOURT WATER SNAKES ( SUBSESSOR BOCOURTI) AND ONE CAPTIVE PUEBLAN MILK SNAKE ( LAMPROPELTIS TRIANGULUM CAMPBELLI).

Two captive Bocourt water snakes ( Subsessor bocourti) presented with chronic white skin lesions on their heads; Ophidiomyces ophiodiicola was identified by culture and polymerase chain reaction (PCR) in skin scrapings from both snakes. Histopathology performed in one Bocourt water snake revealed fungal hyphae in epidermal structures of lesions. One Pueblan milk snake ( Lampropeltis triangulum campbelli) from the same zoologic institution presented with yellow crusts and white blisters on its body, from which O. ophiodiicola was identified by culture and PCR. Two of the three snakes apparently recovered from lesions after multiple natural sheds, whereas the third snake died. This is the first report of O. ophiodiicola infection in Bocourt water snakes and in a Pueblan milk snake, as well as the first report of O. ophiodiicola in France.

DETECTION OF INTRANUCLEAR COCCIDIOSIS IN TORTOISES IN EUROPE AND CHINA.

Intranuclear coccidiosis of tortoises (TINC) has been described in association with systemic disease in various species of tortoises. TINC has been detected in numerous tortoises from the United States, but there are only a few reports from tropical tortoises in Germany and no reports from Asia. Using a real-time polymerase chain reaction assay, samples from 1,011 tortoises were screened for the presence of TINC. Samples originated from animals kept in captivity in Europe and in China. Coccidia were detected in a total of 27 chelonians (2.7%), including the first description of TINC in a marginated tortoise ( Testudo marginata ), Hermann's tortoise ( Testudo hermanni ), African spurred tortoise (Centrochelys sulcata), and yellow-footed tortoise (Chelonoidis denticulatus). The highest percentage of positive animals was found in radiated tortoises ( Astrochelys radiata ). Although the percentage of positive animals was relatively low, this study demonstrates the global distribution of TINC in captive chelonians as well as expanding the known host range for these pathogens.

Ranavirus infections associated with skin lesions in lizards.

Ranaviral disease in amphibians has been studied intensely during the last decade, as associated mass-mortality events are considered to be a global threat to wild animal populations. Several studies have also included other susceptible ectothermic vertebrates (fish and reptiles), but only very few cases of ranavirus infections in lizards have been previously detected. In this study, we focused on clinically suspicious lizards and tested these animals for the presence of ranaviruses. Virological screening of samples from lizards with increased mortality and skin lesions over a course of four years led to the detection of ranaviral infections in seven different groups. Affected species were: brown anoles (Anolis sagrei), Asian glass lizards (Dopasia gracilis), green anoles (Anolis carolinensis), green iguanas (Iguana iguana), and a central bearded dragon (Pogona vitticeps). Purulent to ulcerative-necrotizing dermatitis and hyperkeratosis were diagnosed in pathological examinations. All animals tested positive for the presence of ranavirus by PCR and a part of the major capsid protein (MCP) gene of each virus was sequenced. Three different ranaviruses were isolated in cell culture. The analyzed portions of the MCP gene from each of the five different viruses detected were distinct from one another and were 98.4-100% identical to the corresponding portion of the frog virus 3 (FV3) genome. This is the first description of ranavirus infections in these five lizard species. The similarity in the pathological lesions observed in these different cases indicates that ranaviral infection may be an important differential diagnosis for skin lesions in lizards.