Occurrence of ß-Lactam Resistance Genes and Plasmid-Mediated Resistance Among Vibrios Isolated from Southwest Coast of India.

Antimicrobial resistance (AMR) is a serious global threat driven by the overuse of drugs in humans, animals, as well as the contamination of natural environments with antimicrobial residues. In recent years, the rise of community-acquired infections resistant to antibiotics has drawn renewed attention to the environmental compartment, in particular for pathogens found in aquaculture systems. We quantified the prevalence of antibiotic resistance in Vibrios isolated from the Cochin Estuary as well as the adjoining shrimp farms, and seafood from markets. A total of 280 Vibrio strains were subjected to antimicrobial susceptibility testing and screened for the presence of blaTEM, blaCTX-M, and blaNDM-1 genes. All strains identified were resistant to at least three antimicrobials, and the percentage of drugs resistant per strain ranged from 16% up to 60%. All the strains from the estuary were resistant to amoxicillin, ampicillin, cephalothin, and colistin. Similarly, strains isolated from seafood were resistant to enrofloxacin, furazolidone, and trimethoprim, and all strains from shrimp farms were resistant to colistin. Plasmid-mediated antibiotic resistance was observed in 21% of the strains. In addition, the presence of blaNDM-1 gene was confirmed in 22.85% of the strains. The presence of multiple resistant phenotypes in vibrios, including resistance to last-resort compounds in domestic food sources, raises serious concerns for public health in the Cochin Estuary. Although localized in nature, our findings also have vital implications for the spread of AMR internationally, given the prominence of South India for seafood exports.

GroEL PCR- RFLP - An efficient tool to discriminate closely related pathogenic Vibrio species.

Vibrio sp. are autochthonous to marine and estuarine waters. Several species of Vibrio are pathogens. It is of utmost importance to detect and discriminate the Vibrio sp. that are often involved in food and water borne infections. Since 16S rRNA based identification has limited utility in differentiating the closely related pathogenic species from non pathogenic species, we have evaluated the discriminatory power of groEL PCR-RFLP for identification of closely related Vibrio sp. Accordingly, in the current study, the efficiency of groEL PCR- RFLP for detection and accurate differentiation of known pathogens among Vibrio sp. such as V. cholerae, V. parahaemolyticus, V. vulnificus, V. mimicus, V. fluvialis, V. alginolyticus, V. anguillarum was evaluated. PCR amplified groEL gene fragment of each Vibrio sp. was digested separately using 5 restriction enzymes viz. Hha1, Rsa1, Alu1, Dde1 and Mbo1. The accuracy of the method was further validated by insilico restriction analysis of multiple strains of each species using NEBcutter. The method proved to be efficient for detection and differentiation of Vibrio species under study. Phylogenetic analysis also revealed groEL gene to be a better phylogenetic marker for Vibrio compared to 16S rRNA. Hence, the method can be employed for accurate detection of Vibrio sp. including the closely related species.

Screening and Characterization of Cold-Active ß-Galactosidase Producing Psychrotrophic Enterobacter ludwigii from the Sediments of Arctic Fjord.

Low-temperature-tolerant microorganisms and their cold-active enzymes could be an innovative and invaluable tool in various industrial applications. In the present study, bacterial isolates from the sediment samples of Kongsfjord, Norwegian Arctic, were screened for ß-galactosidase production. Among the isolates, KS25, KS85, KS60, and KS92 have shown good potential in ß-galactosidase production at 20 °C. 16SrRNA gene sequence analysis revealed the relatedness of the isolates to Enterobacter ludwigii. The optimum growth temperature of the isolate was 25 °C. The isolate exhibited good growth and enzyme production at a temperature range of 15-35 °C, pH 5-10. The isolate preferred yeast extract and lactose for the maximum growth and enzyme production at conditions of pH 7.0, temperature of 25 °C, and agitation speed of 100 rpm. The growth and enzyme production was stimulated by Mn2+ and Mg2+ and strongly inhibited by Zn2+, Ni2+, and Cu+. ß-Galactosidases with high specific activity at low temperatures are very beneficial in food industry to compensate the nutritional problem associated with lactose intolerance. The isolate exhibited a remarkable capability to utilize clarified whey, an industrial pollutant, for good biomass and enzyme yield and hence could be well employed in whey bioremediation.

Survival of multi-drug resistant enteropathogenic Escherichia coli and Salmonella paratyphi in Vembanadu lake as a function of saltwater barrier along southwest coast of India.

The objective of the study was to evaluate the survival response of multi-drug resistant enteropathogenic Escherichia coli and Salmonella paratyphi to the salinity fluctuations induced by a saltwater barrier constructed in Vembanadu lake, which separates the lake into a freshwater dominated southern and brackish water dominated northern part. Therefore, microcosms containing freshwater, brackish water and microcosms with different saline concentrations (5, 10, 15, 20, 25 ppt) inoculated with E. coli/S. paratyphi were monitored up to 34 days at 20 and 30 °C. E. coli and S. paratyphi exhibited significantly higher (p < 0.05) survival at 20 °C compared to 30 °C in all microcosms. Despite fresh/brackish water, E. coli and S. paratyphi showed prolonged survival up to 34 days at both temperatures. They also demonstrated better survival potential at all tested saline concentrations except 25 ppt where a significantly higher (p < 0.0001) decay was observed. Therefore, enhanced survival exhibited by the multi-drug resistant enteropathogenic E. coli and S. paratyphi over a wide range of salinity levels suggest that they are able to remain viable for a very long time at higher densities in all seasons of the year in Vembanadu lake irrespective of saline concentrations, and may pose potential public health risks during recreational activities.

An assessment of potential public health risk associated with the extended survival of indicator and pathogenic bacteria in freshwater lake sediments.

Microcosm studies were performed to evaluate the survival of Escherichia coli, Salmonella paratyphi and Vibrio parahaemolyticus in water and sediment collected from the freshwater region of Vembanad Lake (9'35 °N 76'25 °E) along the south west coast of India. All three test microorganisms showed significantly (p < 0.01) higher survival in sediment compared to overlying water. The survival in different sediment types with different particle size and organic carbon content revealed that sediment with small particle size and high organic carbon content could enhance their extended survival (p < 0.05). The results indicate that sediments of the Lake could act as a reservoir of pathogenic bacteria and exhibit a potential health hazard from possible resuspension and subsequent ingestion during recreational activities. Therefore, the assessment of bacterial concentration in freshwater Lake sediments used for contact and non contact recreation has of considerable significance for the proper assessment of microbial pollution of the overlying water, and for the management and protection of related health risk at specific recreational sites. Besides, assessment of the bacterial concentration in sediments can be used as a relatively stable indicator of long term mean bacterial concentration in the water column above.

Isolation and characterization of aerobic thermophilic bacteria from the savusavu hot springs in fiji.

The relative isolation and unique physical properties of the Savusavu Hot Springs in Fiji may yield unique thermophiles. This study was conducted to determine the presence of aerobic thermophilic bacteria in these hot springs. A total of 104 thermophilic bacterial isolates were characterized and using Thermus and Bacillus strains as controls, 58% of the isolates were identified as Anoxybacillus flavithermus, 19% as Geobacillus stearothermophilus/Bacillus licheniformis, 10% as Thermus sp. TG153 and 10% as Thermus sp. TG206. Four isolates were unique in their molecular patterns suggesting there may be novel bacteria in the Savusavu hot springs.

Antibiotic resistance pattern of motile aeromonads from farm raised fresh water fish.

Motile aeromonads isolated from the intestines of farm-raised freshwater fish such as Catla catla, Labeo rohita and Ctenopharyngodon idella have been characterized to species level. Morphological and physiological grouping revealed 61% Aeromonas hydrophila, 30% Aeromonas caviae, 7% Aeromonas sobria and 2% which remained unidentified. Hemolytic activity was detected mostly in A. hydrophila, while only half of the A. sobria and A. caviae showed this activity. Antibiotic resistance patterns of the strains revealed that they had acquired a relatively higher resistance to oxytetracycline, amoxycillin, ampicillin, novobiocin and polymixin-B, implicating possible use of these antibiotics in the aquaculture systems.