User Awareness on ‘Single Window Search’ Tool provided by the KMC Health Sciences Library, MAHE, Manipal: A Study

In academic libraries, information resources are increasing at an alarming rate due to information explosion and development in information and communication technology. Open access resources available for access are also increasing. Academic libraries are putting their best effort to make the process of conducting literature search much easier as users find it difficult to search for literature on a given research topic through several types of resources separately. This necessitated the integration of information resources for easy access. A Web-Scale Discovery tool integrates library subscribed content, library catalogue, digital repositories, open access resources and offers a search and retrieval interface for the users to access the required information easily and quickly. To support users in conducting an effective literature search, KMC Health Sciences Library, MAHE, Manipal has implemented Web-scale Discovery Service, named it as Single Window Search and provided access in Library web portal. The objective of the study was to understand the awareness and satisfaction on different features and modules available at the Single Window Search tool among the library users at KMC Health Sciences Library, MAHE, Manipal. The study revealed that almost all users (92.16%) who participated in the study were aware of the Basic Searching (Title, Keyword, Author) feature of the single window search tool. From the study, it is understood that more than 60.78% of users are satisfied with Basic Searching option and 45.10% of users are satisfied with the Advanced Searching option provided in Single Window Search tool

Quantification of extended-spectrum-beta-lactamase-producing Enterobacteriaceae from water sources in Hall County, Georgia, USA

Extended-spectrum beta-lactamases (ESBLs) are a family of enzymes that confer resistance to a number of antibiotics, including those containing a beta lactam ring. ESBLs exhibit antibiotic resistance by destroying the antibiotic’s structure and may be encoded by bacterial plasmids that can easily be transferred between bacteria from the family Enterobacteriaceae. Organisms that produce ESBLs pose both threats and challenges in the administration of appropriate therapeutic agents to treat infections. Water environments such as streams can help the spread of antibiotic-resistant bacteria which can originate from a variety of sources, including food processing, waste water treatment plants, and urban runoff. We are studying the isolation and identification of ESBL-producing Enterobacteriaceae from water samples obtained from a water treatment plant and its receiving stream in north Georgia. In this paper, we carried out enumeration of ESBL-producing bacteria from water samples obtained immediately upstream and downstream from the water treatment plant. ESBL-producing Enterobacteriaceae were observed from both pre-treated water obtained from the water treatment plant and from upstream (240/ 100 ml) and downstream (240/ 100 ml) water samples, including ESBL-producing Escherichia coli and Klebsiella pneumonia. Our results indicate that ESBL-producing Enterobacteriaceae are present in all water sources sampled. This suggests that the waste water treatment plant is not the source of these microorganisms. Further studies are needed to determine the originating source

The Trichoplusia Ni Gut Microbiome and Its Derivation from the Phyllosphere of Its Food Plants

Insects are the most abundant and diverse animals on planet Earth and rely on their diverse microbiomes to be so. The insect gut microbiome is vital in the growth and development of many insect species. Trichoplusia ni, the cabbage looper, is a generalist herbivore, but little is known about its microbiome. In this study, a metagenomic analysis of fecal samples was used to determine the effect of diet on the microbiome of T. ni larvae. Larvae were reared on six plant species, the microbiome was sampled from fecal material, and the phyllosphere was sampled from leaves the larvae ate. Bacteria were identified using 16S ribosomal RNA gene sequences, and diversity was quantified. Similar phyla, classes, and families were found in both types of samples, suggesting T. ni larvae primarily obtain their gut microbiome from their diet. However, the gut microbiome of T. ni larvae is not identical to the phyllosphere, particularly in relative abundances, suggesting that other factors in the insect gut environment may further modify the diversity of the microbiome. This study adds to the growing body of evidence about the implications of diet for the insect gut microbiome

ANTIBIOTIC-RESISTANT GRAM-NEGATIVE BACTERIA IN A LOCAL WASTEWATER TREATMENT PLANT IN NORTH GEORGIA

Antibiotics are some of the most essential lifesaving drugs currently available to us against infectious diseases. Several pathogens are developing resistance to these drugs and many have become resistant to multiple antibiotics. Such multi-drug resistance is becoming a global issue and has been found in many environments including wastewater treatment plants (WWTPs). In this study, we sampled a local WWTP for antibiotic-resistant bacteria. This WWTP is in North Georgia and runs directly into Lake Lanier, which is the main source of drinking water for the Atlanta area as well as a popular spot for water recreation. We obtained treated and raw sewage water samples. These water samples were filtered, grown in an enrichment media and plated on selective media containing antibiotics that yielded the isolation of antibiotic-resistant bacteria. These isolates were further characterized for antibiotic resistance using disk diffusion method and identified using 16s rRNA gene sequencing. All isolates obtained from both raw sewage and treated water samples were multi-drug resistant. Further analysis using PCR revealed all isolates possessed at least one bla gene and hence were identified to be ESBL-producing bacteria. Our results are consistent with other findings reported worldwide suggesting multi-drug resistant bacteria persist in treated water

Identification and Localization of Myxococcus xanthus Porins and Lipoproteins

Myxococcus xanthus DK1622 contains inner (IM) and outer membranes (OM) separated by a peptidoglycan layer. Integral membrane, β-barrel proteins are found exclusively in the OM where they form pores allowing the passage of nutrients, waste products and signals. One porin, Oar, is required for intercellular communication of the C-signal. An oar mutant produces CsgA but is unable to ripple or stimulate csgA mutants to develop suggesting that it is the channel for C-signaling. Six prediction programs were evaluated for their ability to identify β-barrel proteins. No program was reliable unless the predicted proteins were first parsed using Signal P, Lipo P and TMHMM, after which TMBETA-SVM and TMBETADISC-RBF identified β-barrel proteins most accurately. 228 β-barrel proteins were predicted from among 7331 protein coding regions, representing 3.1% of total genes. Sucrose density gradients were used to separate vegetative cell IM and OM fractions, and LC-MS/MS of OM proteins identified 54 β-barrel proteins. Another class of membrane proteins, the lipoproteins, are anchored in the membrane via a lipid moiety at the N-terminus. 44 OM proteins identified by LC-MS/MS were predicted lipoproteins. Lipoproteins are distributed between the IM, OM and ECM according to an N-terminal sorting sequence that varies among species. Sequence analysis revealed conservation of alanine at the +7 position of mature ECM lipoproteins, lysine at the +2 position of IM lipoproteins, and no noticable conservation within the OM lipoproteins. Site directed mutagenesis and immuno transmission electron microscopy showed that alanine at the +7 position is essential for sorting of the lipoprotein FibA into the ECM. FibA appears at normal levels in the ECM even when a +2 lysine is added to the signal sequence. These results suggest that ECM proteins have a unique method of secretion. It is now possible to target lipoproteins to specific IM, OM and ECM locations by manipulating the amino acid sequence near the +1 cysteine processing site

Survival in Nuclear Waste, Extreme Resistance, and Potential Applications Gleaned from the Genome Sequence of Kineococcus radiotolerans SRS30216

Kineococcus radiotolerans SRS30216 was isolated from a high-level radioactive environment at the Savannah River Site (SRS) and exhibits γ-radiation resistance approaching that of Deinococcus radiodurans. The genome was sequenced by the U.S. Department of Energy's Joint Genome Institute which suggested the existence of three replicons, a 4.76 Mb linear chromosome, a 0.18 Mb linear plasmid, and a 12.92 Kb circular plasmid. Southern hybridization confirmed that the chromosome is linear. The K. radiotolerans genome sequence was examined to learn about the physiology of the organism with regard to ionizing radiation resistance, the potential for bioremediation of nuclear waste, and the dimorphic life cycle. K. radiotolerans may have a unique genetic toolbox for radiation protection as it lacks many of the genes known to confer radiation resistance in D. radiodurans. Additionally, genes involved in the detoxification of reactive oxygen species and the excision repair pathway are overrepresented. K. radiotolerans appears to lack degradation pathways for pervasive soil and groundwater pollutants. However, it can respire on two organic acids found in SRS high-level nuclear waste, formate and oxalate, which promote the survival of cells during prolonged periods of starvation. The dimorphic life cycle involves the production of motile zoospores. The flagellar biosynthesis genes are located on a motility island, though its regulation could not be fully discerned. These results highlight the remarkable ability of K radiotolerans to withstand environmental extremes and suggest that in situ bioremediation of organic complexants from high level radioactive waste may be feasible

Genome Evolution and the Emergence of Fruiting Body Development in Myxococcus xanthus

BACKGROUND: Lateral gene transfer (LGT) is thought to promote speciation in bacteria, though well-defined examples have not been put forward. METHODOLOGY/PRINCIPLE FINDINGS: We examined the evolutionary history of the genes essential for a trait that defines a phylogenetic order, namely fruiting body development of the Myxococcales. Seventy-eight genes that are essential for Myxococcus xanthus development were examined for LGT. About 73% of the genes exhibit a phylogeny similar to that of the 16S rDNA gene and a codon bias consistent with other M. xanthus genes suggesting vertical transmission. About 22% have an altered codon bias and/or phylogeny suggestive of LGT. The remaining 5% are unique. Genes encoding signal production and sensory transduction were more likely to be transmitted vertically with clear examples of duplication and divergence into multigene families. Genes encoding metabolic enzymes were frequently acquired by LGT. Myxobacteria exhibit aerobic respiration unlike most of the delta Proteobacteria. M. xanthus contains a unique electron transport pathway shaped by LGT of genes for succinate dehydrogenase and three cytochrome oxidase complexes. CONCLUSIONS/SIGNIFICANCE: Fruiting body development depends on genes acquired by LGT, particularly those involved in polysaccharide production. We suggest that aerobic growth fostered innovation necessary for development by allowing myxobacteria access to a different gene pool from anaerobic members of the delta Proteobacteria. Habitat destruction and loss of species diversity could restrict the evolution of new bacterial groups by limiting the size of the prospective gene pool

Extended-spectrum Beta -Lactamase Producing Enterobacteriaceae in Flat Creek Water Reclamation Center

Extended-spectrum beta-lactamases (ESBLs) are enzymes that are capable of degrading frequently used antibiotics such as, penicillin and cephalosporin. Organisms that produce these enzymes are therefore multi-drug resistant. ESBLs are commonly produced by the Enterobacteriaceae group of bacteria that are part of normal gut flora. Since first identified in 1893, ESBLs have become increasingly prevalent because ESBL enzymes are plasmid mediated, thus their resistant genes are easily transferred between bacteria via horizontal and vertical gene transmission. Currently, ESBLs pose a significant health threat due to their ability to cause a multitude of difficult to treat infections and are one of the leading causes of death world-wide. As of yet, there is no data on prevalence of ESBL-producing Enterobacteriaceae in North Georgia water sources. Flat creak is located in North Georgia and runs directly into Lake Lanier, which is a main source of drinking water for the Atlanta area as well as a popular spot for water recreation. We obtained efferent and afferent water samples from Flat Creek Water Reclamation Center in Hall County Georgia. These water samples were filtered, grown in an enrichment media and plated on selective media that yielded the isolation of ESBL-producing Enterobacteriaceae. We found ESBL producing enterobacteria in every sample we tested. Further identification methods revealed the presence of ESBL producing Kebsiella pneumoniae, Kebsiella oxytoca, Enterobacter cloacae, Citrobacter freundii and Escherichia coli. Following identification, disk diffusion method was used to test susceptibility of the isolates to several commonly used antibiotics. Lastly, DNA was extracted from the isolates to confirm the presence of bla genes that encode for ESBLs

Multidrug resistance found in extended-spectrum beta-lactamase producing Enterobacteriaceae in Flat Creek Water Reclamation Facility and Flat Creek water samples in Hall County, Georgia.

Extended-spectrum beta-lactamases (ESBLs) are enzymes that destroy several frequently used antibiotics such as, penicillin and cephalosporin. Many pathogenic bacteria harbor these enzymes and are therefore multi-drug resistant. ESBLs are commonly produced by Enterobacteria which are common gut flora of many mammals. ESBLs are plasmid encoded and therefore are easily transferred between bacteria. Because of the rampant and improper use of antibiotics, ESBL producing bacteria have become prevalent and pose a significant health threat due to their ability to cause a multitude of difficult to treat infections. They are one of the leading causes of death world-wide. As of yet, there is no data on prevalence of ESBL-producing Enterobacteriaceae in North Georgia water sources. Flat creak is located in North Georgia and runs directly into Lake Lanier, which is used for swimming and drinking water. We obtained efferent and afferent water samples from Flat Creek Water Reclamation Center in Hall County Georgia. We also collected water samples upstream and downstream from the treatment plant in Flat Creek. These water samples were filtered, grown on an enrichment media and plated on selective media that allowed isolation of ESBL-producing Enterobacteriaceae. We found ESBL producing enterobacteria in all the samples. Further identification methods revealed the presence of ESBL producing Kebsiella pneumoniae, Kebsiella oxytoca, Enterobacter cloacae, and Escherichia coli. Currently we are confirming the presence of bla genes that encode for ESBL in our isolates. We are also carrying out the disk diffusion method to test susceptibility of the isolates to several commonly used antibiotics

13. Bugs in Bugs: Analysis of microbiota of Trichnoplusia ni Larvae

All multicellular organisms including plants and animals harbor a community of microbes called the microbiota. The gut microbiome plays an important role in digestion, immunity and overall health of an animal. Research has shown that the insect microbiome is associated with resistance to natural invaders, virulence, digestion, and reproductive success. The insect microbiome may also metabolize defensive compounds, and influence the ability of insects to associate with certain host plants. Recent research suggests that hornworm caterpillars (Manduca sexta) have no permanent microbiome; instead acquiring transient microbes from the plants they eat. In this project, students from two courses (BIOL1108K and BIOL2500L) collaborated to understand the acquisition of gut microbiome of Trichnoplusia ni, a nocturnal cabbage looper or commonly referred to as owlet moths. Biology 1108K collected fecal samples from T. ni larvae fed on various pre-selected plant diet. The fecal samples were suspended in saline, swabbed on nutrient agar plates, and incubated for 24 h at 37˚C. Bacterial colonies that appeared on these plates were then analyzed and characterized by BIOL2500L students. Our group comprising of four students analyzed three bacterial colonies using microbiological techniques learned in BIOL2500L. We investigated the colony and cell morphology, Gram reaction, antibiotic resistance and biochemical features of all three bacteria. We identified all three bacterial colonies to be Gram-positive Staphylococcus sp, which is interestingly part of the normal skin microbiota suggesting that humans and insects may share similar microbiota