Integron study, molecular typing and characterization of Salmonella serotypes isolated from seafood and poultry

The primary habitat of Salmonella is the gastrointestinal tract of animals and they are discharged into the water bodies through the feces. Aquatic animals act as asymptomatic reservoirs of a wide range of Salmonella serotypes. The inevitable delay in the detection of Salmonella contamination and the low sensitivity of the conventional methods is a serious issue in the seafood industry. Due to the indiscriminate use, the antibiotics are finally accumulated in the aquatic environment which provides the required antibiotic stress for the emergence of more and more antibiotic resistant phenotypes ofSalmonella. Several genetic determinants like integrons, genomic islands etc. play their role in acquisition and reshuffling of antibiotic resistance genes. A large number of virulence determinants are required for Salmonella pathogenicity. The virulence potential of Salmonella is determined, to some extent, by the presence of phages or phage mediated genes in the bacterial genome. There is much intra-serotype polymorphism in Salmonella and epidemiological studies rely on genetic resemblance of the isolated strains. Proper identification of the strain employing the traditional and molecular techniques is a prerequisite for accurate epidemiological studies (Soto et al., 2000). In this context, a study was undertaken to determine the prevalence of different Salmonella serotypes in seafood and to characterize them

Characterization of Bacteriocins from Bacillus licheniformis strain BTHT8 and Bacillus subtilis strain BTFK101 isolated from marine sediment

Pathogenic microorganisms such as Bacillus cereus, Listeria Monocytogenes and Staphylococcus sp have caused serious diseases, and consequently contributed to considerable economic loss in the food and agricultural industries. Antibiotics have been practically used to treat these pathogens since penicillin G was discovered more than half a century ago. Many different types of antibiotics have been discovered or synthesized to control pathogenic microorganisms. Repetitive use and misuse of antibiotics by the agricultural and pharmaceutical industries have caused the emergence of multidrug-resistant microorganisms, even to the strongest antibiotics currently available; therefore, the rapid development of more effective antimicrobial compounds is required to keep pace with demand. Bacteria were isolated from marine water and sediment samples collected from various locations off the coast of Cochin and salt pans of Tuticorin using pour plate technique. One hundred and twelve isolates were obtained. Seventeen isolates exhibiting antimicrobial activity were segregated after primary screening. The secondary screening which was aimed at selection of bacteria that produce proteinaceous inhibitory compounds, helped to select five strains viz. BTFK101, BTHT8, BTKM4, BTEK16 and BTSB22. The five isolates inhibited the growth of six Gram positive test organisms viz. B. cereus, B. circulans, B. coagulans, B. pumilus, Staphylococcus aureus and Clostridium perfringens. After quantitative estimation of the bacteriocin production, the two strains BTFK101 and BTHT8 were selected for further study.

Protease Production by haloarchaea Natrinema sp. BTSH10 isolated from salt pan of South India

The present study led to the recognition of Natrinema sp. BTSH 10 isolated from saltern ponds, as an ideal candidate species for production of gelatinase, which was noted as a halozyme capable of showing enzyme activity in the presence of 15% NaCl. Results obtained during the course of the present study indicated potential for application of this enzyme in industrial catalysis that are performed in the presence of high concentrations of salt. The enzyme characteristics noted with this gelatinase also indicate the scope for probable applications in leather industry, meat tenderization, production of fish sauce and soy sauce. Since halophilic proteases are tolerant to organic solvents, they could be used in antifouling coating preparations used to prevent biofouling of submarine equipments. The gelatinase from haloarchaea could be considered as a probable candidate for peptide synthesis. However, further studies are warranted on this haloarcheal gelatinase particularly on structure elucidation and enzyme engineering to suit a wide range of applications. There is immense scope for developing this halozyme as an industrial enzyme once thorough biochemistry of this gelatinase is studied and a pilot scale study is conducted towards industrial production of this enzyme under fermentation is facilitated. Based on the present study it is concluded that haloarchaea Natrinema sp. that inhabit solar saltern ponds are ideal source for deriving industrially important halozymes and molecular studies on enzymes are prerequisite for their probable industrial applications. This is the first time this species of archaea is recognized as a source of gelatinase enzyme that has potential for industrial applications.

Effect of market-driven minigrids on the frequency stabilization process of isolated middle-sized power grids

Neueste Entwicklungen in Technologien für dezentrale Energieversorgungsstrukturen, erneuerbare Energien, Großhandelsenergiemarkt, Mini- und Mikronetze, verteilte Intelligenz, sowie Informations- und Datenübertragungstechnologien werden die zukünftige Energiewelt maßgeblich bestimmen. Die derzeitigen Forschungsbemühungen zur Vernutzung aller dieser Technologien bilden die Voraussetzungen für ein zukünftiges, intelligentes Stromnetz. Dieses neue Konzept gründet sich auf die folgenden Säulen: Die Versorgung erfolgt durch dezentrale Erzeugungsanlagen und nicht mehr durch große zentrale Erzeuger; die Steuerung beeinflusst nicht mehr allein die Versorgung sondern ermöglich eine auch aktive Führung des Bedarf; die Eingabeparameter des Systems sind nicht mehr nur mechanische oder elektrische Kenngrößen sondern auch Preissignale; die erneuerbaren Energieträger sind nicht mehr nur angeschlossen, sondern voll ins Energienetz integriert. Die vorgelegte Arbeit fügt sich in dieses neue Konzept des intelligenten Stromnetz ein. Da das zukünftige Stromnetz dezentral konfiguriert sein wird, ist eine Übergangsphase notwendig. Dieser Übergang benötigt Technologien, die alle diese neue Konzepte in die derzeitigen Stromnetze integrieren können. Diese Arbeit beweist, dass ein Mininetz in einem Netzabschnitt mittlerer Größe als netzschützende Element wirken kann. Hierfür wurde ein neues Energiemanagementsystem für Mininetze – das CMS (englisch: Cluster Management System) – entwickelt. Diese CMS funktioniert als eine von ökonomischorientierte Betriebsoptimierung und wirkt wie eine intelligente Last auf das System ein, reagierend auf Preissignale. Sobald wird durch eine Frequenzsenkung eine Überlastung des Systems bemerkt, ändert das Mininetz sein Verhalten und regelt seine Belastung, um die Stabilisierung des Hauptnetzes zu unterstützen. Die Wirksamkeit und die Realisierbarkeit des einwickelten Konzept wurde mit Hilfe von Simulationen und erfolgreichen Laborversuchen bewiesen.

Genotyping Klebsiella pneumoniae isolated from hepatic abscesses in three patients from Bogota, Colombia

Pyogenic liver abscess caused by Klebsiella pneumoniae represents an ever increasing entity which has mainly been described as occurring in Asia, even though, on a smaller scale, cases are being more frequently described from the USA and Europe, 13% overall mortality being reached worldwide. Affected patients are severely sick, suffering from fever, sweating, having increased acute phase reactants and risk factors such as Diabetes Mellitus, alcoholism and the inherent characteristics of the bacteria causing the disease. Objective: in this work we used a Multilocus Sequencing Typing (MLST), a nucleotide sequence-based method in order to characterize the genetic relationships among bacterial isolates. Materials and methods: the report is focused on three cases involving patients suffering from pyogenic liver abscess caused by Klebsiella pneumoniae in two hospitals in Bogota, Colombia, where phenotyping and hypermucoviscosity studies were carried out, as well as the genotyping of cultured Klebsiella isolates. Reults: it was found that the isolated microorganism in cases I and II corresponded to the same K. pneumoniae strain, having 100% sequence identity for the 5 genes being studied while the strain in Case III was genotypically different. Conclusion: it is important to carry out multidisciplinary studies allowing all pyogenic liver abscess cases reported in Colombia to be complied to ascertain the frequency of microorganisms causing this pathology in our country, as well as a genotyping study of different K. pneumoniae strains to compare them and confirm clonal and pathogenicity relationships through housekeeping gene analysis.

Differentiation and coordination within a composite training policy providing efficient support namely to isolated museum professionals

In Portugal, namely in municipal museums, the reduced percentage of museum professionals that have a stable administrative position imposes a high degree of polyvalence allowing them to coordinate, often by themselves, the quotidian activities at the museum(s) they run.

Organic farming in isolated landscapes does not benefit flower-visiting insects and pollination

Organic farming has often been found to provide benefits for biodiversity, but the benefits can depend on the species considered and characteristics of the surrounding landscape. In an intensively farmed area of Northeast Italy we investigated whether isolated organic farms, in a conventionally farmed landscape, provided local benefits for insect pollinators and pollination services. We quantified the relative effects of local management (i.e. the farm system), landscape management (proportion of surrounding uncultivated land) and interactions between them. We compared six organic and six conventional vine fields. The proportion of surrounding uncultivated land was calculated for each site at radii of 200, 500, 1000 and 2000 m. The organic fields did not differ from the conventional in their floral resources or proportion of surrounding uncultivated land. Data were collected on pollinator abundance and species richness, visitation rates to, and pollination of experimental potted plants. None of these factors were significantly affected by the farming system. The abundance of visits to the potted plants in the conventional fields tended to be negatively affected by the proportion of surrounding uncultivated land. The proportion fruit set, weight of seeds per plant and seed weight in conventional and organic fields were all negatively affected by the proportion of surrounding uncultivated land. In vine fields the impact of the surrounding landscape was stronger than the local management. Enhancement of biodiversity through organic farming should not be assumed to be ubiquitous, as potential benefits may be offset by the crop type, organicmanagement practices and the specific habitat requirements in the surrounding landscape.

Susceptibility of different insect pupae to the bacterial symbiont, Xenorhabdus nematophila, isolated from the entomopathogenic nematode Steinernema carpocapsae

Cells of the bacterial symbiont Xenorhabdus nematophila from the entomopathogenic nematode, Steinernema carpocapsae entered the pupae of Plutella xylostella after 15 minutes treatment with suspensions containing the bacterial cells. Secretions of Xenorhabdus nematophila, in either broth or water, were found lethal to the pupae of P. xylostella when applied in moist sand. The bacterial symbiont Xenorhabdus nematophila was found lethal to the pupae of greater wax moth (Galleria mellonella), beet armyworm (Spodoptera exigua), diamondback moth (Plutella xylostella) and black vine weevil (Otiorhynchus sulcatus) in the absence of the nematode vector and the cells of X. nematophila entered the haemocoele of the pupae.

Pathogenicity of the bacterium Xenorhabdus nematophila isolated from the entomopathogenic nematode Steinernema carpocapsae and its secretions against Galleria mellonella larvae

The entomopathogenic bacterium, Xenorhabdus nematophila was isolated from the hemolymph of Galleria mellonella infected with Steinernema carpocapsae. The bacterial cells and its metabolic secretions have been found lethal to the Galleria larvae. Toxic secretion in broth caused 95% mortality within 4 d of application whereas the bacterial cells caused 93% mortality after 6 d. When filter and sand substrates were compared, the later one was observed as appropriate. Similarly, bacterial cells and secretion in broth were more effective at 14% moisture and 25 °C temperature treatments. Maximum insect mortality (100%) was observed when bacterial concentration of 4×106 cells/ml was used. Similarly, maximum bacterial cells in broth (95%) were penetrated into the insect body within 2 h of their application. However, when stored bacterial toxic secretion was applied to the insects its efficacy declined. On the other hand, when the same toxic secretion was dried and then dissolved either in broth or water was proved to be effective. The present study showed that the bacterium, X. nematophila or its toxic secretion can be used as an important component of integrated pest management against Galleria.

Proteomic analysis of plasma membrane vesicles isolated from the rat renal cortex

Polarized epithelial cells are responsible for the vectorial transport of solutes and have a key role in maintaining body fluid and electrolyte homeostasis. Such cells contain structurally and functionally distinct plasma membrane domains. Brush border and basolateral membranes of renal and intestinal epithelial cells can be separated using a number of different separation techniques, which allow their different transport functions and receptor expressions to be studied. In this communication, we report a proteomic analysis of these two membrane segments, apical and basolateral, obtained from the rat renal cortex isolated by two different methods: differential centrifugation and free-flow electrophoresis. The study was aimed at assessing the nature of the major proteins isolated by these two separation techniques. Two analytical strategies were used: separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at the protein level or by cation-exchange high-performance liquid chromatography (HPLC) after proteolysis (i.e., at the peptide level). Proteolytic peptides derived from the proteins present in gel pieces or from HPLC fractions after proteolysis were sequenced by on-line liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several hundred proteins were identified in each membrane section. In addition to proteins known to be located at the apical and basolateral membranes, several novel proteins were also identified. In particular, a number of proteins with putative roles in signal transduction were identified in both membranes. To our knowledge, this is the first reported study to try and characterize the membrane proteome of polarized epithelial cells and to provide a data set of the most abundant proteins present in renal proximal tubule cell membranes.

Proteomic analysis of plasma membrane vesicles isolated from the rat renal cortex

Polarized epithelial cells are responsible for the vectorial transport of solutes and have a key role in maintaining body fluid and electrolyte homeostasis. Such cells contain structurally and functionally distinct plasma membrane domains. Brush border and basolateral membranes of renal and intestinal epithelial cells can be separated using a number of different separation techniques, which allow their different transport functions and receptor expressions to be studied. In this communication, we report a proteomic analysis of these two membrane segments, apical and basolateral, obtained from the rat renal cortex isolated by two different methods: differential centrifugation and free-flow electrophoresis. The study was aimed at assessing the nature of the major proteins isolated by these two separation techniques. Two analytical strategies were used: separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at the protein level or by cation-exchange high-performance liquid chromatography (HPLC) after proteolysis (i.e., at the peptide level). Proteolytic peptides derived from the proteins present in gel pieces or from HPLC fractions after proteolysis were sequenced by on-line liquid chromatography-tandem mass spectrometry (LC-MS/MS). Several hundred proteins were identified in each membrane section. In addition to proteins known to be located at the apical and basolateral membranes, several novel proteins were also identified. In particular, a number of proteins with putative roles in signal transduction were identified in both membranes. To our knowledge, this is the first reported study to try and characterize the membrane proteome of polarized epithelial cells and to provide a data set of the most abundant proteins present in renal proximal tubule cell membranes.