Abstract
Aims: This study was undertaken to assess the abundance of multiple-antibiotic-resistant (MAR) Salmonella strains in fecal samples of Rhinoceros unicornis of the Kaziranga National Park (KNP), India.
 Study Design: Antibiotic-resistance profile of the Salmonella isolates from fecal samples of rhinoceros was carried out by replica plating on Muller Hinton Agar (MHA) plates containing antibiotics. The presence of class 1 integrons in metallo-β-lactamase (MBL) producing Salmonella isolates was determined by multiplex PCR.
 Place and Duration of Study: The study was carried out on rhinos of KNP situated in the Golaghat district of Assam, India in April 2015.
 Methodology: Fresh rhino fecal samples (designated as 1R, 2R, 3R, 4R and 5R) were dilution plated onto MacConkey agar. Purified bacterial colonies were then streaked separately on bismuth sulphite (BS) agar plates. All black colonies which are characteric growth of Salmonella were selected and used to make master plates on Luria Agar. To determine the antibiotic-resistance profile of the isolates, master-plates of purified single colonies of Salmonella spp. were replicate-printed on plates containing antibiotics from the β-lactam, aminoglycoside, or quinolone groups. To detect the presence of an integron, a conserved segment polymerase chain reaction was used.
 Results: 97.6% of the Salmonella isolates tested were resistant to >1 antibiotics (multidrug resistant or MAR). A total of 100 isolates from two fecal samples, 4R and 5R, were found to be imipenem resistant; 52 of them tested positive for the presence of MBLs. Five of the twenty MBL producing strains contained class 1 integrons.
 Conclusion: Because Salmonella is usually spread by drinking contaminated water, it is likely that water bodies in KNP are contaminated with MAR Salmonella strains. In case of Salmonellosis outbreak among Indian one-horned rhinos, our antibiogram will assist the veterinarians to choose the appropriate regimen of antibiotics for the rhinos in the KNP.
Highlights
The greater Indian one-horned rhinoceros (Rhinoceros unicornis) is mostly confined to the Kaziranga National Park (KNP) in Assam's Golaghat district
Salmonellosis caused by Salmonella spp. infection in both black and Indian one-horned rhinoceroses (Diceros bicornis, Rhinoceros unicornis) is often fatal [7]
We found that the incidence and abundance of MAR Salmonella spp. were high in fecal samples from Indian rhinos, and that a significant proportion of MAR isolates carried class 1 integrons
Summary
The greater Indian one-horned rhinoceros (Rhinoceros unicornis) is mostly confined to the Kaziranga National Park (KNP) in Assam's Golaghat district. Bian and his colleagues [5] investigated the gut microbiota of the white rhinoceros They used barcoded pyrosequencing to characterize 105,651 16S rRNA gene sequences obtained from fecal samples from five white rhinoceroses. Salmonellosis caused by Salmonella spp. infection in both black and Indian one-horned rhinoceroses (Diceros bicornis, Rhinoceros unicornis) is often fatal [7]. Many pathogenic Salmonella isolates have integrons containing antibiotic-resistant gene cassettes. In 2003, Lindstedt and his colleagues [12] found integrons with sizes 650, 1000, 1200, 1500, 1600, 1700, 2000 and 2100 bp in many of the isolates of S. typhimurium (97 %) and S. enteritidis (22%) Such isolates, pose a threat as a zoonotic pathogen with increased resistance to several antibiotics. Yong and colleagues [14] identified clinical isolates of Pseudomonas spp. and Acinetobacter spp. that produce metallo-βlactamase (MBL)
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