Characterization of L-asparaginase from Streptomyces koyangensis SK4 with acrylamide-minimizing potential in potato chips.

Microbial L-asparaginase is well known for its application in food industries to reduce acrylamide content in fried starchy food. L-asparaginase produced by Arctic actinomycetes Streptomyces koyangensis SK4 was purified and studied for biochemical characterization. The L-asparaginase was purified with a yield of 15.49% and final specific activity of 179.77 IU/mg of protein. The enzyme exhibited a molecular weight of 43 kDa. The optimum pH and temperature for maximum activity of the purified enzyme were 8.5 °C and 40 °C, respectively. The enzyme expressed maximum activity at an incubation period of 30 min and a substrate concentration of 0.06 M. The enzyme has a low Km value of 0.041 M and excellent substrate specificity toward L-asparagine. The enzyme activity was inhibited by metal ions Ba2+ and Hg2+, while Mn2+ and Mg2+ enhanced the activity. The study evaluated the acrylamide reduction potential of L-asparaginase from Streptomyces koyangensis SK4 in potato chips. The blanching plus L-asparaginase treatment of potato slices resulted in a 50% reduction in acrylamide content. The study illustrated an effective acrylamide reduction strategy in potato chips using L-asparaginase from a psychrophilic actinomycete. Besides the acrylamide reduction potential, L-asparaginase from Streptomyces koyangensis SK4 also did not exhibit any glutaminase or urease activity which is an outstanding feature of L-asparaginase to be used as a chemotherapeutic agent.

Dissemination of multidrug resistant bacteria to the polar environment - Role of the longest migratory bird Arctic tern (Sterna paradisaea).

The ever-increasing prevalence of antibiotic-resistant bacteria(ARB), primarily due to the frequent use and misuse of antibiotics, is an issue of serious global concern. Migratory birds have a significant role in dissemination of ARB, as they acquire resistant bacteria from reservoirs and transport them to other environments which are relatively less influenced by anthropogenically. We have investigated the prevalence of ARB in a long-distance migratory bird, the Arctic tern (Sterna paradisaea) captured from the Svalbard Archipelago. The birds were tagged with geolocators to track their extraordinary long migration, and the cloacal samples were collected before the migration and after the migration by recapturing the same birds. The tracking of 12 birds revealed that during the annual cycle they underwent a total of 166 stopovers (11-18, mean = 3.8) and recovery points along the Atlantic Ocean. Twelve major bacterial genera were identified from Arctic tern cloacal samples, which are dominated by Staphylococcus spp. and Aerococcus spp. The bacterial isolates showed resistance against 16 antibiotics (before migration) and 17 antibiotics (after migration) out of 17 antibiotics tested. Resistance to ß-lactam and quinolone class of antibiotics were frequent among the bacteria. The study highlights the potential role of Arctic tern in the dissemination of multidrug resistant bacteria across far and wide destinations, especially to the polar environments.

Bacterial community structure and functional profiling of high Arctic fjord sediments.

Kongsfjorden, an Arctic fjord is significantly affected by the glacier melt and Atlantification, both the processes driven by accelerated warming in the Arctic. This has lead to changes in primary production, carbon pool and microbial communities, especially that in the sediment. In this study, we have examined the bacterial community structure of surface (0-2 cm) and subsurface (3-9 cm) sediments of Kongsfjorden using the high throughput sequencing analysis. Results revealed that bacterial community structure of Kongsfjorden sediments were dominated by phylum Proteobacteria followed by Bacteroidetes and Epsilonbacteraeota. While α- and γ-Proteobacterial class were dominant in surface sediments; δ-Proteobacteria were found to be predominant in subsurface sediments. The bacterial community structure in the surface and subsurface sediments showed significant variations (p ≤ 0.05). Total organic carbon could be one of the major parameters controlling the bacterial diversity in the surface and subsurface sediments. Functional prediction analysis indicated that the bacterial community could be involved in the degradation of complex organic compounds such as glycans, glycosaminoglycans, polycyclic aromatic hydrocarbons and also in the biosynthesis of secondary metabolites.

Influence of environmental factors on benthic nitrogen fixation and role of sulfur reducing diazotrophs in a eutrophic tropical estuary.

Benthic nitrogen fixation in the tropical estuaries plays a major role in marine nitrogen cycle, its contribution to nitrogen budget and players behind process is not well understood. The present study was estimated the benthic nitrogen fixation rate in a tropical estuary (Cochin) and also evaluated the contribution of various diazotrophic bacterial communities. Nitrogen fixation was detected throughout year (0.1-1.11 nmol N g-1 h-1); higher activity was observed in post-monsoon. The nifH gene abundance was varied from 0.8 × 104 to 0.6 × 108 copies g-1dry sediment; highest was detected in post-monsoon. The Cluster I and Cluster III were the dominant diazotrophs. Sulfur reducing bacterial phylotypes (Deltaproteobacteria) contributed up to 2-72% of total nitrogen fixation. These bacteria may provide new nitrogen to these systems, counteracting nitrogen loss via denitrification and anammox. Overall, the study explained the importance of benthic nitrogen fixation and role of diazotrophs in a monsoon influenced tropical estuarine environments.

Abundance and diversity of diazotrophs in the surface sediments of Kongsfjorden, an Arctic fjord.

Diazotrophy in the Arctic environment is poorly understood compared to tropical and subtropical regions. Hence in this study, we report the abundance and diversity of diazotrophs in Arctic fjord sediments and elucidate the role of environmental factors on the distribution of diazotrophs. The study was conducted during the boreal summer in the Kongsfjorden, an Arctic fjord situated in the western coast of Spitsbergen. The abundance of nifH gene was measured through quantitative real-time PCR and the diversity of diazotrophs was assessed by nifH targeted clone library and next generation sequence analysis. Results revealed that the abundance of nifH gene in the surface sediments ranged from 2.3 × 106 to 3.7 × 107 copies g- 1. The δ-proteobacterial diazotrophs (71% of total sequence) were the dominant class observed in this study. Major genera retrieved from the sequence analysis were Desulfovibrionaceae (25% of total sequence), Desulfuromonadaceae (18% of total sequence) and Desulfobacteriaceae (10% of total sequence); these are important diazotrophic iron and sulfur-reducing bacterial clade in the Kongsfjorden sediments. The abundance of nifH gene showed a significant positive correlation TOC/TN ratio (r2 = 0.96, p ≤ 0.05) and total organic carbon (p ≤ 0.05) content in the fjord sediments. The higher TOC/TN ratio (4.24-14.5) indicated low nitrogen content organic matter in the fjord sediments through glacier runoff, which enhances the abundance and diversity of nitrogen fixing microorganisms.

Further report of Bariaka alopiae Cressey, 1966 (Copepoda, Siphonostomatoida) from the Indian Ocean with new host and geographic record.

A copepod parasite, Bariaka alopiae Wilson, 1932 (Eudactylinidae) infested on thresher sharks caught from the Indian Exclusive Economic Zone off Andaman and Nicobar archipelago, eastern Indian Ocean is further reported. Adult females of B. alopiae were collected from the gill filaments of two host species, Alopias pelagicus Nakamura and A. superciliosus Lowe. The species B. alopiae can be easily distinguished from other species within the genus by the following characteristic features: cylindrical body devoid of spines, eighteen segmented antennules and four segmented abdomen. In the Indian Ocean, this parasite was known only from its original description 51 years ago, based on materials from western Indian Ocean off Madagascar. Further the present study reports, A. pelagicus as a new host for this parasite.

Heavy-metal resistance in Gram-negative bacteria isolated from Kongsfjord, Arctic.

Isolation and characterization of heterotrophic Gram-negative bacteria was carried out from the sediment and water samples collected from Kongsfjord, Arctic. In this study, the potential of Arctic bacteria to tolerate heavy metals that are of ecological significance to the Arctic (selenium (Se), mercury (Hg), cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn)) was investigated. Quantitative assay of 130 isolates by means of plate diffusion and tube dilution methods was carried out by incorporation of different concentrations of metals. Growth in Se and Pb at a concentration of 3000 µg/L was significantly lower (P≤0.0001) than at 2000 µg/L. The minimum inhibitory concentration for Cd and Hg was 50 µg/L (P≤0.0001, F=264.23 and P≤0.0001, F=291.08, respectively) even though in the tube dilution test, Hg-containing tubes showed much less growth, revealing its superior toxicity to Cd. Thus, the level of toxicity of heavy metals was found to be in the order of Hg>Cd>Cu>Zn>Pb>Se. Multiple-metal-resistant isolates were investigated for their resistance against antibiotics, and a positive correlation was observed between antibiotic and metal resistance for all the isolates tested. The resistant organisms thus observed might influence the organic and inorganic cycles in the Arctic and affect the ecosystem.

An investigation into occasional White Spot Syndrome Virus outbreak in traditional paddy cum prawn fields in India.

A yearlong (September 2009-August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved oxygen (DO) content of water from feeder canal showed low oxygen levels (as low as 0.8 mg/L) throughout the study period. There was no disease outbreak in the perennial ponds. Poor water exchange coupled with nutrient loading from adjacent houses resulted in phytoplankton bloom in shallow seasonal ponds which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon besides considerably high alkaline pH. Ammonia levels were found to be very high in these ponds. WSSV outbreak was encountered twice during the study leading to mass mortalities in the seasonal ponds. The hypoxia and high ammonia content in water and abrupt fluctuations in temperature, salinity and pH might lead to considerable stress in the shrimps triggering WSSV infection in these traditional ponds.

Relative survival of Escherichia coli and Salmonella typhimurium in a tropical estuary.

Microcosm studies have been carried out to find out the relative survival of Escherichia coli and Salmonella typhimurium in a tropical estuary. Survival has been assessed in relation to the important self-purifying parameters such as biotic factors contained in the estuarine water, toxicity due to the dissolved organic and antibiotic substances in the water and the sunlight. The results revealed that sunlight is the most important inactivating factor on the survival of E. coli and S. typhimurium in the estuarine water. While the biological factors contained in the estuarine water such as protozoans and bacteriophages also exerted considerable inactivation of these organisms, the composition of the water with all its dissolved organic and inorganic substances was not damaging to the test organisms. Results also indicated better survival capacity of E. coli cells under all test conditions when compared to S. typhimurium.

Microbial quality of shrimp products of export trade produced from aquacultured shrimp.

Bacteriological quality of individually quick frozen (IQF) shrimp products produced from aquacultured tiger shrimp (Penaeus monodon) has been analysed in terms of aerobic plate count (APC), coliforms, Escherichia coli, coagulase-positive staphylococci, Salmonella, and Listeria monocytogenes. Eight hundred forty-six samples of raw, peeled, and deveined tail-on (RPTO), 928 samples of cooked, peeled, and deveined tail-on (CPTO), 295 samples of headless, undeveined shell-on (HLSO), and 141 samples of raw, peeled, and deveined tail-off (RPND) shrimps were analysed for the above bacteriological parameters. Salmonella was isolated in only one sample of raw, peeled tail-on. Serotyping of the strain revealed that it was S. typhimurium. While none of the cooked, peeled tail-on shrimp samples exceeded the aerobic plate count (APC) of 10(5) colony forming units per gram (cfu/g), 2.5% of raw, peeled, tail-on, 6.4% of raw, peeled tail-off, and 7.5% of headless shell-on shrimp samples exceeded that level. Coliforms were detected in all the products, though at a low level. Prevalence of coliforms was higher in headless shell-on (26%) shrimps followed by raw, peeled, and deveined tail-off (19%), raw, peeled tail-on (10%), and cooked, peeled tail-on (3.8%) shrimps. While none of the cooked, peeled tail-on shrimp samples were positive for coagulase-positive staphylococci and E. coli, 0.6-1.3% of the raw, peeled tail-on were positive for staphylococci and E. coli, respectively. Prevalence of staphylococci was highest in raw, peeled tail-off (5%) shrimps and the highest prevalence of E. coli (4.8%) was noticed in headless shell-on shrimps. L. monocytogenes was not detected in any of the cooked, peeled tail-on shrimps. Overall results revealed that the plant under investigation had exerted good process control in order to maintain superior bacteriological quality of their products.