Tsukamurella serpentis sp. nov., isolated from the oral cavity of Chinese cobras (Naja atra).

Two bacterial strains, HKU54T and HKU55, were isolated from the oral cavity of two Chinese cobras (Naja atra) in Hong Kong. 16S rRNA gene sequence analysis revealed 100 % sequence identity between HKU54T and HKU55, and the two strains shared 99.0 % sequence identities with Tsukamurella inchonensis ATCC 700082T. The two strains had unique biochemical profiles distinguishable from closely related species of the genus Tsukamurella. DNA-DNA hybridization confirmed that they belonged to the same species (≥92.1±7.9 % DNA-DNA relatedness) but were distinct from all other known species of the genus Tsukamurella (≤52.6±5.3 % DNA-DNA relatedness). Chemotaxonomic and morphological analyses of the two strains also demonstrated results consistent with their classification in the genus Tsukamurella. The DNA G+C contents of strains HKU54T and HKU55 were 69.2±1.5 mol% and 69.2±1.3 mol% (mean±sd; n=3) respectively. A novel species, Tsukamurella serpentis sp. nov., is proposed to accommodate strains HKU54T and HKU55, with HKU54T (=JCM 31017T=DSM 100915T) designated as the type strain.

Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina.

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina.

Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina. / Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina.

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89

) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina.

Comparison of different protocols for the extraction of microbial DNA from reef corals

This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

Salmonella serovar distribution in cobras (Naja kaouthia), snake-food species, and farm workers at Queen Saovabha Snake Park, Thailand.

The purpose of the current study was to investigate the prevalence and serovar distribution of Salmonella isolates in cobras and their environment at a snake park. A total of 166 fecal or intestinal samples were examined, comprising 39 samples from captive cobras (Naja kaouthia), 70 from recently wild-caught cobras, 19 from wild-caught cobras that had been kept on the farm for over 3 months, 18 from mice (Mus musculus), 12 from frogs (Hoplobatrachus rugulosus), and 8 from farm workers. Specific serological identification was performed, and pulsed-field gel electrophoresis (PFGE) was utilized for DNA analysis. Out of all snakes (n = 128), 20 of the 30 animals used for snake food and 3 of the 8 samples from personnel were positive for Salmonella spp. There were 228 Salmonella isolates, with a total of 29 serovars from subspecies I and IIIb, composed of 24 serovars from cobras and 5 from the other sources. Salmonella Amsterdam was predominant in captive-born and captive cobras, followed by S. Poona and S. Bareilly, respectively (P < 0.05). Salmonella I 4,[5],12:i:- was the sole serovar detected from the mice, while 3 serovars including Ramatgan, I 4,[5],12:e,h:-, and rough strain were detected only from frogs (P < 0.001). Salmonella Derby was only detected in workers. On the basis of the PFGE results, evidence of movement of isolates between human beings and snakes, and between snakes and frogs, was found for S. Poona and S. Wandsworth, respectively. The study suggests that Salmonella spp. act as true residents in the intestinal tract of cobras with high risk of environmental contamination through fecal shedding.

Oral bacterial flora of the Chinese cobra (Naja atra) and bamboo pit viper (Trimeresurus albolabris) in Hong Kong SAR, China.

OBJECTIVE: To determine the oral bacterial flora associated with two common local venomous snakes in Hong Kong, namely the Chinese cobra (Naja atra) and the bamboo pit viper (Trimeresurus albolabris). DESIGN: Cross-sectional study. SETTING: A non-government organisation and a regional hospital in Hong Kong. SUBJECTS: Thirty-two Chinese cobras and seven bamboo pit vipers. MAIN OUTCOME MEASURES: Species identification of bacteria in the oral cavity of both snakes and their antibiotic susceptibilities. RESULTS: The oral cavity of Chinese cobra harbour a wide range of pathogenic bacteria, including: Gram-negative bacterial species like Morganella morganii, Aeromonas hydrophila and Proteus, and Gram-positive bacteria like Enterococcus faecalis, coagulase-negative Staphylococcus as well as anaerobic species (clostridia). The oral cavity of the Chinese cobra is more likely than that of the bamboo pit viper to harbour pathogenic bacteria associated with snakebite infection (P<0.001). The median number of pathogenic bacteria per snake was significantly higher in the Chinese cobra (P<0.001). All pathogenic Gram-negative bacteria isolated were susceptible to levofloxacin. Amoxicillin/clavulanate provided good coverage against pathogenic Gram-positive bacteria (Enterococcus faecalis) and anaerobes. CONCLUSION: 'Prophylactic' antibiotic treatment for Chinese cobra bites may be beneficial, owing to the multiple pathogenic bacteria in its oral cavity and the higher risk of ensuing necrosis. The regimen of levofloxacin plus amoxicillin/clavulanate appears promising for this purpose, but further study is required to confirm its clinical utility in patients.

Dermatophilus chelonae in a king cobra (Ophiophagus hannah).

A mass was removed from the left flank of a 10-yr-old male king cobra (Ophiophagus hannah), and histologic examination revealed granulomatous dermatitis with intralesional gram-positive cocci and filamentous bacteria. Fourteen months later, a histologically similar subcutaneous mass was removed from a different site. One year later, a large subcutaneous mass at the first surgical site was removed, and histopathologic examination revealed multiloculated granulomas with intralesional gram-positive cocci. An organism was cultured and identified by 16S ribosomal RNA gene sequencing as Dermatophilus chelonae. After a course of antibiotic therapy, no further lesions were seen for 5 mo.