Diversity and relative abundance of the bacterial pathogen, Flavobacterium spp., infecting reproductive ecotypes of kokanee salmon.

Science.gov (United States)

Lemay, Matthew A; Russello, Michael A

2014-11-04

Understanding the distribution and abundance of pathogens can provide insight into the evolution and ecology of their host species. Previous research in kokanee, the freshwater form of sockeye salmon (Oncorhynchus nerka), found evidence that populations spawning in streams may experience a greater pathogen load compared with populations that spawn on beaches. In this study we tested for differences in the abundance and diversity of the gram-negative bacteria, Flavobacterium spp., infecting tissues of kokanee in both of these spawning habitats (streams and beaches). Molecular assays were carried out using primers designed to amplify a ~200 nucleotide region of the gene encoding the ATP synthase alpha subunit (AtpA) within the genus Flavobacterium. Using a combination of DNA sequencing and quantitative PCR (qPCR) we compared the diversity and relative abundance of Flavobacterium AtpA amplicons present in DNA extracted from tissue samples of kokanee collected from each spawning habitat. We identified 10 Flavobacterium AtpA haplotypes among the tissues of stream-spawning kokanee and seven haplotypes among the tissues of beach-spawning kokanee, with only two haplotypes shared between spawning habitats. Haplotypes occurring in the same clade as F. psychrophilum were the most prevalent (92% of all reads, 60% of all haplotypes), and occurred in kokanee from both spawning habitats (streams and beaches). Subsequent qPCR assays did not find any significant difference in the relative abundance of Flavobacterium AtpA amplicons between samples from the different spawning habitats. We confirmed the presence of Flavobacterium spp. in both spawning habitats and found weak evidence for increased Flavobacterium diversity in kokanee sampled from stream-spawning sites. However, the quantity of Flavobacterium DNA did not differ between spawning habitats. We recommend further study aimed at quantifying pathogen diversity and abundance in population-level samples of kokanee combined with

Fish mucus alters the Flavobacterium columnare transcriptome

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Columnaris disease which is caused by Flavobacterium columnare severely impacts the production of freshwater finfish species. Due to the impact on the aquaculture industry, research efforts to better understand the biological processes of F. columnare including the formation of biofilms and their co...

The zebrafish galectins Drgal1-L2 and Drgal3-L1 bind in vitro to the infectious hematopoietic necrosis virus (IHNV) glycoprotein and reduce viral adhesion to fish epithelial cells

Digital Repository Service at National Institute of Oceanography (India)

Nita-Lazar, M.; Mancini, J.; Feng, C.; Gonzalez-Montalban, N.; Ravindran, C.; Jackson, S.; Heras-Sanchez, A.D.L.; Giomarelli, B.; Ahmed, H.; Haslam, S.M.; Wu, G.; Dell, A.; Ammayappan, A.; Vakharia, V.N.; Vasta, G.R.

galectin-3 (Drgal3-L1) in IHNV adhesion to epithelial cells. Our results suggest that the extracellular Drgal1-L2 and Drgal3-L1 interact directly and in a carbohydrate-dependent manner with the IHNV glycosylated envelope and glycans on the epithelial cell...

Modulation of genes related to the recruitment of immune cells in the digestive tract of trout experimentally infected with infectious pancreatic necrosis virus (IPNV) or orally vaccinated.

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Ballesteros, Natalia A; Rodríguez Saint-Jean, Sylvia; Pérez-Prieto, Sara I; Aquilino, Carolina; Tafalla, Carolina

2014-05-01

There are still many details of how intestinal immunity is regulated that remain unsolved in teleost. Although leukocytes are present all along the digestive tract, most immunological studies have focused on the posterior segments and the importance of each gut segment in terms of immunity has barely been addressed. In the current work, we have studied the regulation of several immune genes along five segments of the rainbow trout (Oncorhynchus mykiss) digestive tract, comparing the effects observed in response to an infectious pancreatic necrosis virus (IPNV) infection to those elicited by oral vaccination with a plasmid coding for viral VP2. We have focused on the regulation of several mucosal chemokines, chemokine receptors, the major histocompatibility complex II (MHC-II) and tumor necrosis factor α (TNF-α). Furthermore, the recruitment of IgM(+) cells and CD3(+) cells was evaluated along the different segments in response to IPNV by immunohistochemical techniques. Our results provide evidences that there is a differential regulation of these immune genes in response to both stimuli along the gut segments. Along with this chemokine and chemokine receptor induction, IPNV provoked a mobilization of IgM(+) and IgT(+) cells to the foregut and pyloric caeca region, and CD3(+) cells to the pyloric caeca and midgut/hindgut regions. Our results will contribute to a better understanding of how mucosal immunity is orchestrated in the different gut segments of teleost. Copyright © 2013 Elsevier Ltd. All rights reserved.

Evaluation of an Experimental Re-introduction of Sockeye Salmon into Skaha Lake; Year 2 of 3, 2001 Technical Report.

Energy Technology Data Exchange (ETDEWEB)

Fisher, Christopher (Colville Confederated Tribes, Omak Community Center, Omak, WA); Machin, Deanna; Wright, Howie (Okanagan National Fisheries Commission, Westbank, BC, Canada)

2002-04-01

This report summarizes the findings from YEAR 2 of a three-year disease risk assessment. The Okanagan Nation Fisheries Commission (ONFC) and the Colville Confederated Tribes (CCT) are investigating the risks involved in re-introducing sockeye salmon into Skaha Lake, part of their historical range (Ernst and Vedan 2000). The disease risk assessment compares the disease and infection status of fish above and below McIntyre Dam (the present limit of sockeye migration). The disease agents identified that are of a particular concern are: infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus type 2 (IHNV type2), erythrocytic inclusion body syndrome virus (EIBSV), the whirling disease agent (Myxobolus cerebralis), and the ceratomyxosis agent (Ceratomyxa shasta).

Flavobacterium kingsejongi sp. nov., a carotenoid-producing species isolated from Antarctic penguin faeces.

Science.gov (United States)

Choi, Jun Young; Kim, Jin Ho; Lee, Pyung Cheon

2018-03-01

Taxonomic studies were carried out on a carotenoid-producing strain, designated WV39 T , isolated from the faeces of Antarctic penguins. Cells of strain WV39 T were Gram-stain-negative, strictly aerobic, yellow and rod-shaped. 16S rRNA gene sequence analysis revealed that strain WV39 T was closely related to Flavobacterium qiangtangense JCM 19739 T (96.3 % similarity), Flavobacterium noncentrifugens NBRC 108844 T (95.5 %) and Flavobacterium aquatile LMG 4008 T (94.9 %). The predominant cellular fatty acids were iso-C15 : 0, iso-C15 : 0 3-OH and summed feature 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1ω7c). Menaquinone-6 was the sole quinone identified, and the major pigment was zeaxanthin. The major polar lipid was phosphatidylethanolamine. DNA-DNA relatedness of strain WV39 T with respect to its closest phylogenetic neighbours was 41.8 % for F. qiangtangense JCM 19739 T , 25.5 % for F. aquatile LMG 4008 T and 25.2 % for F. noncentrifugens NBRC 108844 T . The DNA G+C content of strain WV39 T was 39.8 mol%. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain WV39 T is concluded to represent a novel species of the genus Flavobacterium, for which the name Flavobacteriumkingsejongi sp. nov. is proposed. The type strain is WV39 T (=KCTC 42908 T =CECT 9085 T ).

Flavobacterium meningosepticum sepsis in an infant with a diarrheal prodrome.

Science.gov (United States)

Springer, S C; Johnson, G M

1999-02-01

A full term, previously normal 2 1/2-month-old black boy was transferred to our hospital from an outlying facility on hospital day 5 for failure to thrive. Three weeks before transfer, the infant was hospitalized for a diarrheal illness with fever. The baby received 3 days of ceftriaxone empirically and was discharged home after the sepsis evaluation was negative. Mild diarrhea and steady weight loss continued and the baby was readmitted. Blood culture done on admission grew Flavobacterium meningosepticum, an organism previously described as an uncommon cause of sepsis in neonates and immunocompromised individuals. As it is water-borne, it has been associated with infection via contaminated water. This organism is usually resistant to antibiotics commonly used for empiric treatment. To our knowledge, this is the first reported case of Flavobacterium bacteremia associated with a prodromal and concurrent diarrheal illness.

Flavobacterium hibisci sp. nov., isolated from the rhizosphere of Hibiscus syriacus L.

Science.gov (United States)

Moya, Gabriela; Yan, Zheng-Fei; Trinh, Huan; Won, Kyung-Hwa; Yang, Jung-Eun; Kook, Moo-Chang; Yi, Tae-Hoo

2017-04-01

A Gram-stain-negative, smooth, bright-yellow-pigmented, rod-shaped bacterial strain, slightly motile by gliding, catalase- and oxidase-positive and aerobic, but growing weakly under anaerobic conditions, was isolated from the rhizosphere of the flower mugunghwa (Hibiscus syriacus L.) located in Kyung Hee University, Yongin, Gyeonggi, South Korea. The strain named THG-HG1.4T grew at 15-35 °C, pH 6.5-9.0 and in the presence of 0-2.5 % (w/v) NaCl. The phylogenetic analysis based on 16S rRNA gene sequence showed that strain THG-HG1.4T was most closely related to Flavobacterium gyeonganense HME7524T (98.83 %) and Flavobacterium arsenitoxidans S2-3HT (97.28 %). The DNA G+C content of strain THG-HG1.4T was 41.2 mol%. In DNA-DNA hybridization, the DNA-DNA relatedness between strain THG-HG1.4T and its closest phylogenetic neighbour was below 64.1 %. The predominant isoprenoid quinone detected in strain THG-HG1.4T was menaquinone-6 (MK-6). The major polar lipids were found to be phosphatidylethanolamine, three unidentified lipids, two unidentified glycolipids and an unidentified aminolipid. The major fatty acids were identified as iso-C15 : 0, iso-C15 : 0 3-OH, C16 : 0, iso-C17 : 0 3-OH and summed feature 3. Thus, based on the report of the phenotypic, genotypic and phylogenetic characterization of strain THG-HG1.4T, it has been concluded that the novel isolate represents a novel species of the genus Flavobacterium.Flavobacterium hibisci sp. nov. is proposed, with THG-HG1.4T (=KACC 18852T=CCTCC AB 2016178T) as the type strain.

Flavobacterium psychrophilum, prevention and immune response

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Dalsgaard, Inger

The fish pathogen Flavobacterium psychrophilum is one of the main causes of mortality in farmed rainbow trout and other salmonid fish. The disease following infection is often called bacterial coldwater disease (BCWD) in USA or rainbow trout fry syndrome (RTFS) in Europe. An infected farm can exp...... goal is to examine gene expression and location of transcription products in rainbow trout fry, in order to optimize vaccination or immune-stimulation. The presentation will focus on the future plans for the project, since no data have yet been obtained....

Isolation and characterization of an Antarctic Flavobacterium strain with agarase and alginate lyase activities

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Lavín Paris

2016-09-01

Full Text Available Several bacteria that are associated with macroalgae can use phycocolloids as a carbon source. Strain INACH002, isolated from decomposing Porphyra (Rhodophyta, in King George Island, Antarctica, was screened and characterized for the ability to produce agarase and alginate-lyase enzymatic activities. Our strain INACH002 was identified as a member of the genus Flavobacterium, closely related to Flavobacterium faecale, using 16S rRNA gene analysis. The INACH002 strain was characterized as psychrotrophic due to its optimal temperature (17ºC and maximum temperature (20°C of growth. Agarase and alginate-lyase displayed enzymatic activities within a range of 10°C to 50°C, with differences in the optimal temperature to hydrolyze agar (50°C, agarose (50°C and alginate (30°C during the first 30 min of activity. Strain Flavobacterium INACH002 is a promising Antarctic biotechnological resource; however, further research is required to illustrate the structural and functional bases of the enzymatic performance observed during the degradation of different substrates at different temperatures.

Genetic and serological typing of European infectious haematopoietic necrosis virus (IHNV) isolates

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Johansson, T.; Einer-Jensen, K.; Batts, W.; Ahrens, P.; Bjorkblom, C.; Kurath, G.; Bjorklund, H.; Lorenzen, N.

2009-01-01

Infectious haematopoietic necrosis virus (IHNV) causes the lethal disease infectious haematopoietic necrosis (IHN) in juvenile salmon and trout. The nucleocapsid (N) protein gene and partial glycoprotein (G) gene (nucleotides 457 to 1061) of the European isolates IT-217A, FR-32/87, DE-DF 13/98 11621, DE-DF 4/99-8/99, AU-9695338 and RU-FR1 were sequenced and compared with IHNV isolates from the North American genogroups U, M and L. In phylogenetic studies the N gene of the Italian, French, German and Austrian isolates clustered in the M genogroup, though in a different subgroup than the isolates from the USA. Analyses of the partial G gene of these European isolates clustered them in the M genogroup close to the root while the Russian isolate clustered in the U genogroup. The European isolates together with US-WRAC and US-Col-80 were also tested in an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (MAbs) against the N protein. MAbs 136-1 and 136-3 reacted equally at all concentrations with the isolates tested, indicating that these antibodies identify a common epitope. MAb 34D3 separated the M and L genogroup isolates from the U genogroup isolate. MAb 1DW14D divided the European isolates into 2 groups. MAb 1DW14D reacted more strongly with DE-DF 13/98 11621 and RU-FR1 than with IT-217A, FR- 32/87, DE-DF 4/99-8/99 and AU-9695338. In the phylogenetic studies, the Italian, French, German and Austrian isolates clustered in the M genogroup, whereas in the serological studies using MAbs, the European M genogroup isolates could not be placed in the same specific group. These results indicate that genotypic and serotypic classification do not correlate. ?? 2009 Inter-Research.

Sickeningly sweet: L-rhamnose stimulates Flavobacterium columnare biofilm formation and virulence

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Flavobacterium columnare, the causative agent of columnaris disease causes substantial mortality worldwide in numerous freshwater finfish species. Due to its global significance and impact on the aquaculture industry continual efforts to better understand basic mechanisms that contribute to disease ...

Phenotypic and genotypic characterization of Flavobacterium psychrophilum from Finnish fish farms

DEFF Research Database (Denmark)

Madetoja, J.; Hanninen, M.L.; Hirvela-Koski, V.

2001-01-01

Finnish isolates (n = 37) of Flavobacterium psychrophilum isolated from farmed salmonids were studied using phenotypic and genotypic characteristics. The characteristics of isolates were compared with the characteristics of Swedish and Estonian F. psychrophilum isolates and the type strain, F...

Virus isolation vs RT-PCR: which method is more successful in detecting VHSV and IHNV in fish tissue sampled under field conditions?

DEFF Research Database (Denmark)

Knüsel, R.; Bergmann, S. M.; Einer-Jensen, Katja

2007-01-01

in Switzerland. Compared to SPNT, the RT-PCR method detected, as with virus isolation, a much lower number of positive cases; reasons for this discrepancy are discussed. Our results indicate that RT-PCR can not only be successfully applied in field surveys, but may also be slightly more sensitive than virus......This study compared the results of reverse transcription-polymerase chain reaction (RT-PCR) and traditional virus isolation on cell culture in detection of viral haemorrhagic septicaemia virus (VHSV) and infectious haematopoietic necrosis virus (IHNV). RT-PCR was used for 172 tissue sample pools...... (total of 859 fish) originating from a field survey on the occurrence of VHSV and IHNV in farmed and wild salmonids in Switzerland. These samples represented all sites with fish that were either identified as virus-positive by means of virus isolation (three sites, four positive tissue sample pools) and...

Comparative analysis of the Flavobacterium columnare genomovar I and II genomes

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Columnaris disease caused by Gram-negative rod Flavobacterium columnare is one of the most common diseases of catfish. F. columnare is also a common problem in other cultured fish species worldwide. F. columnare has three major genomovars; we have sequenced a representative strain from genomovar I (...

Use of ribosomal proteins as biomarkers for identification of Flavobacterium psychrophilum by MALDI-TOF mass spectrometry.

Science.gov (United States)

Fernández-Álvarez, Clara; Torres-Corral, Yolanda; Santos, Ysabel

2018-01-06

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) is a rapid methodology for identification of bacteria that is increasingly used in diagnostic laboratories. This work aimed at evaluating the potential of MALDI-TOF-MS for identification of the main serotypes of Flavobacterium psychrophilum isolated from salmonids, and its discrimination from closely related Flavobacterium spp. A mass spectra library was constructed by analysing 70 F. psychrophilum strains representing the serotypes O1, O2a, O2b and O3, including reference and clinical isolates. Peak mass lists were examined using the Mass-Up software for the detection of potential biomarkers, similarity and cluster analysis. Fourteen species-identifying biomarkers were detected in all the F. psychrophilum isolates tested, moreover, sets of serotype-identifying biomarkers ions were selected. F. psychrophilum-specific biomarkers were identified as ribosomal proteins by matching with protein databases. Furthermore, sequence variation corresponding to amino acid exchanges in several biomarker proteins were tentatively assigned. Closely related Flavobacterium species (F. flevense, F. succinicans, F. columnare, F. branchiophilum and F. johnsoniae) could be differentiated from F. psychrophilum by defining species identifying biomarkers and hierarchical cluster analysis. These results demonstrated that MALDI-TOF spectrometry represents a powerful tool for an accurate identification of the fish pathogen F. psychrophilum as well as for epidemiological studies. The results obtained in this study demonstrated that MALDI-TOF mass spectrometry represents a powerful tool that can be used by diagnostic laboratories for rapid identification of the fish pathogen Flavobacterium psychrophilum and its differentiation from other Flavobacterium-related species. Analysis of mass peak lists revealed the potential of the MALDI-TOF technique to identify epidemiologically important serotypes affecting

The carbohydrate L-rhamnose promotes biofilm formation which enhances Flavobacterium columnare virulence

Science.gov (United States)

Flavobacterium columnare, the causative agent of columnaris disease causes substantial mortality worldwide in numerous freshwater finfish species. Due to its global significance and impact on the aquaculture industry, continual efforts to better understand basic mechanisms that contribute to disease...

Commercial demonstration of kaolinitic clay for protection of Flavobacterium columnaris in sportfish

Science.gov (United States)

Sportfish farms in Arkansas routinely battle Columnaris disease, which is caused by Flavobacterium columnare. Columnaris is especially prevalent during the feed training of centrarchids such as largemouth bass and immediately following harvest of crappie, redear sunfish, and bluegill while they are ...

Influence of native catfish mucus on Flavobacterium columnare growth and proteolytic activity

Science.gov (United States)

Flavobacterium columnare causes columnaris disease of farmed and wild freshwater fish. Skin mucus is an important factor in early stages of columnaris pathogenesis, albeit little studied. Our objectives were to 1) characterize the terminal glycosylation pattern (TGP) of catfish mucus, 2) determine t...

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002 from four tropical fish species in Brazil

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F. Pilarski

Full Text Available Flavobacterium columnare is the causative agent of columnaris disease in freshwater fish, implicated in skin and gill disease, often causing high mortality. The aim of this study was the isolation and characterization of Flavobacterium columnare in tropical fish in Brazil. Piracanjuba (Brycon orbignyanus, pacu (Piaractus mesopotamicus, tambaqui (Colossoma macropomum and cascudo (Hypostomus plecostomus were examined for external lesions showing signs of colunmaris disease such as greyish white spots, especially on the head, dorsal part and caudal fin of the fish. The sampling comprised 50 samples representing four different fish species selected for study. Samples for culture were obtained by skin and kidney scrapes with a sterile cotton swabs of columnaris disease fish and streaked onto Carlson and Pacha (1968 artificial culture medium (broth and solid which were used for isolation. The strains in the liquid medium were Gram negative, long, filamentous, exhibited flexing movements (gliding motility, contained a large number of long slender bacteria and gathered into ‘columns'. Strains on the agar produced yellow-pale colonies, rather small, flat that had rhizoid edges. A total of four Flavobacterium columnare were isolated: 01 Brycon orbignyanus strain, 01 Piaractus mesopotamicus strain, 01 Colossoma macropomum strain, and 01 Hypostomus plecostomus strain. Biochemical characterization, with its absorption of Congo red dye, production of flexirubin-type pigments, H2S production and reduction of nitrates proved that the isolate could be classified as Flavobacterium columnare.

Survival of Flavobacterium psychrophilum in rainbow trout ( Oncorhynchus mykiss ) serum in vitro

DEFF Research Database (Denmark)

Wiklund, T.; Dalsgaard, Inger

2002-01-01

Virulent and non-virulent strains of Flavobacterium psychrophilum of different serotypes were examined for survival and growth in non-immune and immune rainbow trout serum, in vitro. A majority of the examined strains consumed complement of non-immune serum, but the complement cascade was not able...

An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either Piscirickettsia salmonis or IPNV

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Bols Niels C

2010-04-01

Full Text Available Abstract Background Due to the limited number of species specific antibodies against fish proteins, differential gene expression analyses are vital for the study of host immune responses. Quantitative real-time reverse transcription PCR (qRT-PCR is one of the most powerful tools for this purpose. Nevertheless, the accuracy of the method will depend on the careful selection of genes whose expression are stable and can be used as internal controls for a particular experimental setting. Findings The expression stability of five commonly used housekeeping genes [beta-actin (ACTB, elongation factor 1-alpha (EF1A, ubiquitin (UBQ, glyceraldehyd-3-phosphate dehydrogenase (GAPDH and tubulin alpha (TUBA] were monitored in salmonid cell lines CHSE-214 and RTS11 after infection with two of the most fastidious fish pathogens, the facultative bacterium Piscirickettsia salmonis and the aquabirnavirus IPNV (Infectious Pancreatic Necrosis Virus. After geNorm analysis, UBQ and EF1A appeared as the most stable, although EF1A was slightly upregulated at late stages of P. salmonis infection in RTS11. ACTB instead, showed a good performance in each case, being always considered within the three most stable genes of the panel. In contrast, infection-dependent differential regulation of GAPDH and TUBA was also demonstrated. Conclusion Based on the data presented here with the cell culture models CHSE-214 and RTS11, we suggest the initial choice of UBQ, ACTB and EF1A as reference genes in qRT-PCR assays for studying the effect of P. salmonis and IPNV on the host immune response.

Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve.

Science.gov (United States)

Wadowsky, R M; Yee, R B

1983-12-01

Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae. Both isolates were recovered from the hot water tanks of hospitals. Ferric PPi stimulated satellite growth between 0.01 and 0.1%.

Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve.

OpenAIRE

Wadowsky, R M; Yee, R B

1983-01-01

Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae. Both isolates were recovered from the hot water tanks of hospitals. Ferric PPi stimulated satellite growth between 0.01 and 0.1%.

Identification of four distinct phylogenetic groups in Flavobacterium columnare with fish host associations

Science.gov (United States)

Columnaris disease, caused by the Gram-negative bacterium Flavobacterium columnare, is one of the most prevalent fish diseases worldwide. An exceptionally high level of genetic diversity among isolates of F. columnare has long been recognized, whereby six established genomovars have been described t...

Early viral replication and induced or constitutive immunity in rainbow trout families with differential resistance to Infectious hematopoietic necrosis virus (IHNV)

Science.gov (United States)

Purcell, M.K.; LaPatra, S.E.; Woodson, J.C.; Kurath, G.; Winton, J.R.

2010-01-01

The main objective of this study was to assess correlates of innate resistance in rainbow trout full-sibling families that differ in susceptibility to Infectious hematopoietic necrosis virus (IHNV). As part of a commercial breeding program, full-sibling families were challenged with IHNV by waterborne exposure at the 1 g size to determine susceptibility to IHNV. Progeny from select families (N = 7 families) that varied in susceptibility (ranging from 32 to 90% cumulative percent mortality (CPM)) were challenged again at the 10 g size by intra-peritoneal injection and overall mortality, early viral replication and immune responses were evaluated. Mortality challenges included 20–40 fish per family while viral replication and immune response studies included 6 fish per family at each time point (24, 48 and 72 h post-infection (hpi)). CPM at the 1 g size was significantly correlated with CPM at the 10 g size, indicating that inherent resistance was a stable trait irrespective of size. In the larger fish, viral load was measured by quantitative reverse-transcriptase PCR in the anterior kidney and was a significant predictor of family disease outcome at 48 hpi. Type I interferon (IFN) transcript levels were significantly correlated with an individual's viral load at 48 and 72 hpi, while type II IFN gene expression was significantly correlated with an individual's viral load at 24 and 48 hpi. Mean family type I but not type II IFN gene expression was weakly associated with susceptibility at 72 hpi. There was no association between mean family susceptibility and the constitutive expression of a range of innate immune genes (e.g. type I and II IFN pathway genes, cytokine and viral recognition receptor genes). The majority of survivors from the challenge had detectable serum neutralizing antibody titers but no trend was observed among families. This result suggests that even the most resistant families experienced sufficient levels of viral replication to trigger specific

More Than Gliding: Involvement of GldD and GldG in the Virulence of Flavobacterium psychrophilum

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David Pérez-Pascual

2017-11-01

Full Text Available A fascinating characteristic of most members of the genus Flavobacterium is their ability to move over surfaces by gliding motility. Flavobacterium psychrophilum, an important pathogen of farmed salmonids worldwide, contains in its genome the 19 gld and spr genes shown to be required for gliding or spreading in Flavobacterium johnsoniae; however, their relative role in its lifestyle remains unknown. In order to address this issue, two spreading deficient mutants were produced as part of a Tn4351 mutant library in F. psychrophilum strain THCO2-90. The transposons were inserted in gldD and gldG genes. While the wild-type strain is proficient in adhesion, biofilm formation and displays strong proteolytic activity, both mutants lost these characteristics. Extracellular proteome comparisons revealed important modifications for both mutants, with a significant reduction of the amounts of proteins likely transported through the outer membrane by the Type IX secretion system, indicating that GldD and GldG proteins are required for an effective activity of this system. In addition, a significant decrease in virulence was observed using rainbow trout bath and injection infection models. Our results reveal additional roles of gldD and gldG genes that are likely of importance for the F. psychrophilum lifestyle, including virulence.

Evaluation of the antibody response to the LV-359-01 strain of flavobacterium columnare

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Flavobacterium columnare, the causative agent of columnaris disease produces substantial mortality worldwide among numerous freshwater farmed finfish species. As aquaculture production continues to increase the frequency of columnaris disease will only continue to rise. Add to this an increase in re...

Effectiveness of copper sulfate and potassium permanganate on channel catfish infected with Flavobacterium columnare

Science.gov (United States)

Copper sulfate (CuSO4) and potassium permanganate (KMnO4) were evaluated for their effectiveness to curtail mortality and decrease bacterial load in fish tissues and water in channel catfish Ictalurus punctatus naturally infected with Flavobacterium columnare, the causative agent of columnaris. Fis...

Draft genome sequence of the fish pathogen Flavobacterium columnare strain CSF-298-10

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We announce the genome assembly of Flavobacterium columnare strain CSF-298-10, a strain isolated from an outbreak of Columnaris disease at a commercial trout farm in Snake River Valley Idaho, USA. The complete genome consists of 13 contigs totaling 3,284,579 bp, average G+C content of 31.5% and 2933...

Immunization of rainbow trout Oncorhynchus mykiss (Walbaum) with a crude lipopolysaccharide extract from Flavobacterium psychrophilum

Science.gov (United States)

Control methods for Flavobacterium psychrophilum, the etiologic agent of bacterial coldwater disease (CWD) and rainbow trout fry syndrome, are limited and oftentimes ineffective; hence, research efforts have focused on vaccine development. This study tested the hypothesis that a crude lipopolysacch...

Utilization of oligo- and polysaccharides at microgram-per-litre levels in freshwater by Flavobacterium johnsoniae

NARCIS (Netherlands)

Sack, E.L.W.; Wielen, van der P.W.J.J.; Kooij, van der D.

2010-01-01

Aims: To obtain a bacterial strain that can be used to quantify biodegradable polysaccharides at concentrations of a few micrograms per litre in freshwater. Methods and Results: Flavobacterium johnsoniae strain A3 was isolated from tap water supplemented with laminarin, pectin or amylopectin at 100

Variable Number of Tandem Repeats (VNTR) analysis of Flavobacterium psychrophilum from salmonids in Chile and Norway

DEFF Research Database (Denmark)

Apablaza, Patricia; Brevik, Oyvind J.; Mjos, Svein

2015-01-01

Background: Flavobacterium psychrophilum causes serious fish diseases such RTFS and BCWD, affecting the aquaculture industry worldwide. Commercial vaccines are not available and control of the disease depends on the use of antibiotics. Reliable methods for detection and identification of differen...

Reproducible challenge model to investigate the virulence of Flavobacterium columnare genomovars in rainbow trout Oncorhynchus mykiss

Science.gov (United States)

Flavobacterium columnare is a Gram-negative bacterium that causes columnaris disease and has significant economic impacts on aquaculture production worldwide. Molecular analyses have demonstrated that there is genetic diversity among F. columnare isolates. A review of the published literature that u...

Genomic characterization of Flavobacterium psychrophilum serotypes and development of a multiplex PCR-based serotyping scheme

DEFF Research Database (Denmark)

Rochat, Tatiana; Fujiwara-Nagata, Erina; Calvez, Ségolène

2017-01-01

Flavobacterium psychrophilum is a devastating bacterial pathogen of salmonids reared in freshwater worldwide. So far, serological diversity between isolates has been described but the underlying molecular factors remain unknown. By combining complete genome sequence analysis and the serotyping me...... for bacterial coldwater disease resistance and future vaccine formulation....

Quality control ranges for testing broth microdilution susceptibility of Flavobacterium columnare and F. psychrophilium to nine antimicrobials

Science.gov (United States)

A multi-laboratory broth microdilution method trial was performed to standardize the specialized test conditions required for fish pathogens Flavobacterium columnare and F. pyschrophilum. Nine laboratories tested the quality control (QC) strains Escherichia coli ATCC 25922 and Aeromonas salmonicid...

Evaluation of potassium permanganate against an experimental subacute infection of Flavobacterium columnare in channel catfish, Icatlurus punctatus

Science.gov (United States)

The efficacy of potassium permanganate (KMnO4) as a prophylactic and therapeutic treatment for subacute infection of Flavobacterium columnare was demonstrated in experimentally infected channel catfish, Ictalurus punctatus. Catfish experimentally infected with F. columnare to mimic a subacute infec...

A comparative study on phyllosphere nitrogen fixation by newly isolated Corynebacterium sp. & Flavobacterium sp. and their potentialities as biofertilizer.

Science.gov (United States)

Giri, S; Pati, B R

2004-01-01

A number of nitrogen fixing bacteria has been isolated from forest phyllosphere on the basis of nitrogenase activity. Among them two best isolates are selected and identified as Corynebacterium sp. AN1 & Flavobacterium sp. TK2 able to reduce 88 and 132 n mol of acetylene (10(8)cells(-1)h(-1)) respectively. They were grown in large amount and sprayed on the phyllosphere of maize plants as a substitute for nitrogenous fertilizer. Marked improvements in growth and total nitrogen content of the plant have been observed by the application of these nitrogen-fixing bacteria. An average 30-37% increase in yield was obtained, which is nearer to chemical fertilizer treatment. Comparatively better effect was obtained by application of Flavobacterium sp.

Flavobacterium psychrophilum, invasion into and shedding by rainbow trout Oncorhynchus mykiss

DEFF Research Database (Denmark)

Madetoja, J.; Nyman, P.; Wiklund, T.

2000-01-01

The infection route of Flavobacterium psychrophilum into rainbow trout Oncorhynchus mykiss was studied using bath and cohabitation challenges as well as oral challenge with live feed as a vector. Additionally, the number of bacterial cells shed by infected fish into the surrounding water...... is discussed as an important invasion route for F. psychrophilum into the fish. The shedding rate of F. psychrophiIlun by infected fish was associated with water temperature and the mortality of the infected fish. High numbers of F. psychrophilum cells were released into the water by dead rainbow trout during...

Effect of Bacteriophages on the Growth of Flavobacterium psychrophilum and Development of Phage-Resistant Strains

DEFF Research Database (Denmark)

Christiansen, Rói Hammershaimb; Madsen, Lone; Dalsgaard, Inger

2016-01-01

The controlling effect of single and multiple phages on the density of Flavobacterium psychrophilum at different initial multiplicity of infection (MOI) was assessed in batch cultures to explore the potential for phage-based treatment of this important fish pathogen. A high initial phage concentr......The controlling effect of single and multiple phages on the density of Flavobacterium psychrophilum at different initial multiplicity of infection (MOI) was assessed in batch cultures to explore the potential for phage-based treatment of this important fish pathogen. A high initial phage...... concentration (MOI = 0.3–4) was crucial for efficient viral lysis, resulting in a 104–105-fold reduction of phage-sensitive cells (both single phages and phage cocktails), which was maintained throughout the incubation (>10 days). Following cell lysis, regrowth of phage-resistant strains was examined...... and resistant strains were isolated for further characterization. The application of a mathematical model allowed simulation of phage-host interactions and resistance development, confirming indications from strain isolations that phage-sensitive strains dominated the regrowing population (>99.8 %) at low MOI...

Occurrence of Flavobacterium psychrophilum in fish-farming environments

DEFF Research Database (Denmark)

Madetoja, J.; Dalsgaard, Inger; Wiklund, T.

2002-01-01

Occurrence of Flavobacterium psychrophilum in fish farms and fish-farming environments was studied using agar plate cultivation,the immunoflourescence antibody technique (IFAT) and nested PCR. Characteristics of 64 F. psychrophilum isolates from rainbow trout Oncorhynchus mykiss, fish farm rearing....... F. psychrophilum was detected and isolated from skin mucus, skin lesions and internal organs of diseased rainbow trout and from fish without clinical disease. The pathogen was also present in wild perch Perca fluviatilis, roach Rutilus rutilus, and ovarian fluids of farmed rainbow trout brood fish...... ribopatterns), ribotype A was the most dominant. Farmed rainbow trout brood fish carried a broad-spectrum of serologically and genetically different F. psychrophilum in ovarian fluids. Virulence of the tested isolates in rainbow trout varied and naturally infected rainbow trout shed 104 to 108 cells fish-1 h-1...

Composition of Extracellular Polymeric Substances (EPS) produced by Flavobacterium columnare isolated from tropical fish in Brazil.

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de Alexandre Sebastião, Fernanda; Pilarski, Fabiana; Lemos, Manoel Victor Franco

2013-01-01

Thirty nine isolates of Flavobacterium columnare from Brazilian fish farms had their carbohydrate composition of EPS evaluated by high efficiency liquid chromatography, using the phenol-sulfuric acid method of EPS. The occurrence of capsules on F. columnare cells was not directly related to biofilm formation, and the predominant monosaccharide is glucose.

Identification of genes encoding the type IX secretion system and secreted proteins in Flavobacterium columnare IA-S-4

Science.gov (United States)

Flavobacterium columnare, a member of the phylum Bacteroidetes, causes columnaris disease in wild and aquaculture-reared freshwater fish. The mechanisms responsible for columnaris disease are not known. Many members of the phylum Bacteroidetes use type IX secretion systems (T9SSs) to secrete enzymes...

Competition of a parathion-hydrolyzing Flavobacterium with bacteria from ditch water in carbon-, nitrate- and phosphate-limited continuous cultures

NARCIS (Netherlands)

Sprenger, W.; Dijkstra, A.; Zwart, G.; Van Agterveld, M.P.; Van Noort, P.C.M.; Parsons, J.R.

2003-01-01

The effect of competition for macroelements with bacteria from ditch water on the parathion-hydrolyzing Flavobacterium sp. ATCC 27551 (FB) was investigated within mixed continuous cultures under carbon-, nitrate- or phosphate-limited conditions. The high initial rate of parathion hydrolysis

Comparative effects of copper sulfate or potassium permanganate on channel catfish concurrently infected with Flavobacterium columnare and Ichthyobodo necator

Science.gov (United States)

An opportunistic study was conducted to determine the effects of two chemical therapeutants on channel catfish (CCF) Ictalurus punctatus concurrently infected Flavobacterium columnare and Ichthyobodo necator. Copper sulfate (CuSO4) and potassium permanganate (KMnO4) were investigated for their abil...

In vitro comparisons of the inhibitory activity of florfenicol copper sulfate and potassium permanganate towards Aeromonas hydrophila and Flavobacterium columnare

Science.gov (United States)

Aeromonas hydrophila and Flavobacterium columnare, the etiological agents of motile aeromonas septicemia (MAS) and columnaris disease, respectively, have been recently causing crippling moralities to the sunshine bass, Morone chrysops female X Morone saxatilis male (Percichthyidae), industry in the ...

Development of a reproducible challenge model to investigate the virulence of Flavobacterium columnare genomovars in rainbow trout Oncorhynchus mykiss (Walbaum)

Science.gov (United States)

Flavobacterium columnare is a Gram-negative bacterium that causes columnaris disease and has significant economic impacts on aquaculture production worldwide. Molecular analyses have demonstrated that there is genetic diversity among F. columnare isolates. A review of the published literature that...

Experimental Transmission of Infectious Pancreatic Necrosis Virus from the Blue Mussel, Mytilus edulis, to Cohabitating Atlantic Salmon (Salmo salar) Smolts

Science.gov (United States)

Pietrak, Michael R.; Bricknell, Ian

2013-01-01

Integrated multitrophic aquaculture (IMTA) reduces the environmental impacts of commercial aquaculture systems by combining the cultivation of fed species with extractive species. Shellfish play a critical role in IMTA systems by filter-feeding particulate-bound organic nutrients. As bioaccumulating organisms, shellfish may also increase disease risk on farms by serving as reservoirs for important finfish pathogens such as infectious pancreatic necrosis virus (IPNV). The ability of the blue mussel (Mytilus edulis) to bioaccumulate and transmit IPNV to naive Atlantic salmon (Salmo salar) smolts was investigated. To determine the ability of mussels to filter and accumulate viable IPNV, mussels were held in water containing log 4.6 50% tissue culture infective dose(s) (TCID50) of the West Buxton strain of IPNV ml−1. Viable IPNV was detected in the digestive glands (DGs) of IPNV-exposed mussels as early as 2 h postexposure. The viral load in mussel DG tissue significantly increased with time and reached log 5.35 ± 0.25 TCID50 g of DG tissue−1 after 120 h of exposure. IPNV titers never reached levels that were significantly greater than that in the water. Viable IPNV was detected in mussel feces out to 7 days postdepuration, and the virus persisted in DG tissues for at least 18 days of depuration. To determine whether IPNV can be transmitted from mussels to Atlantic salmon, IPNV-exposed mussels were cohabitated with naive Atlantic salmon smolts. Transmission of IPNV did occur from mussels to smolts at a low frequency. The results demonstrate that a nonenveloped virus, such as IPNV, can accumulate in mussels and be transferred to naive fish. PMID:23872575

Epidemiological cut-off values for Flavobacterium psychrophilum MIC data generated by a standard test protocol.

Science.gov (United States)

Smith, P; Endris, R; Kronvall, G; Thomas, V; Verner-Jeffreys, D; Wilhelm, C; Dalsgaard, I

2016-02-01

Epidemiological cut-off values were developed for application to antibiotic susceptibility data for Flavobacterium psychrophilum generated by standard CLSI test protocols. The MIC values for ten antibiotic agents against Flavobacterium psychrophilum were determined in two laboratories. For five antibiotics, the data sets were of sufficient quality and quantity to allow the setting of valid epidemiological cut-off values. For these agents, the cut-off values, calculated by the application of the statistically based normalized resistance interpretation method, were ≤16 mg L(-1) for erythromycin, ≤2 mg L(-1) for florfenicol, ≤0.025 mg L(-1) for oxolinic acid (OXO), ≤0.125 mg L(-1) for oxytetracycline and ≤20 (1/19) mg L(-1) for trimethoprim/sulphamethoxazole. For ampicillin and amoxicillin, the majority of putative wild-type observations were 'off scale', and therefore, statistically valid cut-off values could not be calculated. For ormetoprim/sulphadimethoxine, the data were excessively diverse and a valid cut-off could not be determined. For flumequine, the putative wild-type data were extremely skewed, and for enrofloxacin, there was inadequate separation in the MIC values for putative wild-type and non-wild-type strains. It is argued that the adoption of OXO as a class representative for the quinolone group would be a valid method of determining susceptibilities to these agents. © 2014 John Wiley & Sons Ltd.

Evaluation of the therapeutic effect of potassium permanganate at early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus)

Science.gov (United States)

The efficacy of potassium permanganate (KMnO4) against early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus) was evaluated. Fish were experimentally challenged, by waterborne exposure for 2 h to F. columnare after cutaneous abrasion, an...

PKR Activation Favors Infectious Pancreatic Necrosis Virus Replication in Infected Cells

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Amr A.A. Gamil

2016-06-01

Full Text Available The double-stranded RNA-activated protein kinase R (PKR is a Type I interferon (IFN stimulated gene that has important biological and immunological functions. In viral infections, in general, PKR inhibits or promotes viral replication, but PKR-IPNV interaction has not been previously studied. We investigated the involvement of PKR during infectious pancreatic necrosis virus (IPNV infection using a custom-made rabbit antiserum and the PKR inhibitor C16. Reactivity of the antiserum to PKR in CHSE-214 cells was confirmed after IFNα treatment giving an increased protein level. IPNV infection alone did not give increased PKR levels by Western blot, while pre-treatment with PKR inhibitor before IPNV infection gave decreased eukaryotic initiation factor 2-alpha (eIF2α phosphorylation. This suggests that PKR, despite not being upregulated, is involved in eIF2α phosphorylation during IPNV infection. PKR inhibitor pre-treatment resulted in decreased virus titers, extra- and intracellularly, concomitant with reduction of cells with compromised membranes in IPNV-permissive cell lines. These findings suggest that IPNV uses PKR activation to promote virus replication in infected cells.

A simple and efficient method for the extraction and separation of menaquinone homologs from wet biomass of Flavobacterium.

Science.gov (United States)

Wei, Hongfei; Zhao, Genhai; Liu, Hui; Wang, Han; Ni, Wenfeng; Wang, Peng; Zheng, Zhiming

2018-01-01

Menaquinone homologs (MK-n), that is, MK-4, MK-5, and MK-6, can be produced by the fermentation of Flavobacterium. In this study, we proposed a simple and efficient method for the extraction of menaquinones from wet cells without the process of drying the biomass. Meanwhile, a rapid and effective solution for the separation of menaquinone homologs was developed using a single organic solvent, which was conducive to the recovery of the solvent. The results showed that the highest yield was obtained with pretreatment using absolute ethanol at a ratio of 6:1 (v/m) for 30 min and then two extractions of 30 min each using methanol at a ratio of 6:1 (v/m). The recovery efficiency of the menaquinones reached to 102.8% compared to the positive control. The menaquinone homologs were effectively separated using methanol as eluent at a flow rate of 0.52 mL/min by a glass reverse-phase C18 silica gel column with a height-to-diameter ratio of 5.5:1. The recovery of menaquinones achieved was 99.6%. In conclusion, the methods for extraction and separation of menaquinone homologs from wet Flavobacterium cells were simple and efficient, which makes them suitable not only on a laboratory scale but also for application on a large scale.

Genomic structure of bacteriophage 6H and its distribution as prophage in Flavobacterium psychrophilum strains

DEFF Research Database (Denmark)

Castillo Bermúdez, Daniel Elías; Espejo, Romilio; Middelboe, Mathias

2014-01-01

Flavobacterium psychrophilum is currently one of the most devastating fish pathogens worldwide causing considerable economic losses in salmonid aquaculture. Recently, attention has been drawn to the use of phages for controlling F. psychrophilum, and phages infecting the pathogen have been isolated...... showed > 80% amino acid similarity to a specific region found in the virulent F. psychrophilum strain JIP02/86 (ATCC 49511), suggesting that a prophage similar to phage 6H was present in this strain. Screening for a collection of 49 F. psychrophilum strains isolated in Chile, Denmark, and USA...

A nuclear localization of the infectious haematopoietic necrosis virus NV protein is necessary for optimal viral growth.

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Myeong Kyu Choi

Full Text Available The nonvirion (NV protein of infectious hematopoietic necrosis virus (IHNV has been previously reported to be essential for efficient growth and pathogenicity of IHNV. However, little is known about the mechanism by which the NV supports the viral growth. In this study, cellular localization of NV and its role in IHNV growth in host cells was investigated. Through transient transfection in RTG-2 cells of NV fused to green fluorescent protein (GFP, a nuclear localization of NV was demonstrated. Deletion analyses showed that the (32EGDL(35 residues were essential for nuclear localization of NV protein, and fusion of these 4 amino acids to GFP directed its transport to the nucleus. We generated a recombinant IHNV, rIHNV-NV-ΔEGDL in which the (32EGDL(35 was deleted from the NV. rIHNVs with wild-type NV (rIHNV-NV or with the NV gene replaced with GFP (rIHNV-ΔNV-GFP were used as controls. RTG-2 cells infected with rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL yielded 12- and 5-fold less infectious virion, respectively, than wild type rIHNV-infected cells at 48 h post-infection (p.i.. While treatment with poly I∶C at 24 h p.i. did not inhibit replication of wild-type rIHNVs, replication rates of rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL were inhibited by poly I∶C. In addition, both rIHNV-ΔNV and rIHNV-NV-ΔEGDL induced higher levels of expressions of both IFN1 and Mx1 than wild-type rIHNV. These data suggest that the IHNV NV may support the growth of IHNV through inhibition of the INF system and the amino acid residues of (32EGDL(35 responsible for nuclear localization are important for the inhibitory activity of NV.

Effect of hydrogen peroxide and/or Flavobacterium psychrophilum on the gills of rainbow trout, Oncorhynchus mykiss (Walbaum)

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Kania, P. W.; Buchmann, K.

2015-01-01

The immune response and morphological changes in the gills of rainbow trout fry after immersion in hydrogen peroxide (H2O2), Flavobacterium psychrophilum or combined exposure were examined. The gills were sampled 4, 48, 125 and 192 h after exposure, and the regulation of expression of the following...... with gene expression. Although pretreatment with H2O2 before immersion in F. psychrophilum did not significantly alter the amount of bacteria found in the gill, the immune response was influenced: exposure to F. psychrophilum resulted in a negative correlation with expression of IL‐17c1, MHC I and MHC II...

Evaluation of the immune response in rainbow trout fry, Oncorhynchus mykiss (Walbaum), after waterborne exposure to Flavobacterium psychrophilum and/or hydrogen peroxide

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Kania, P. W.; Buchmann, Kurt

2015-01-01

The immune response in rainbow trout fry against Flavobacterium psychrophilum was elucidated using an immersion-based challenge with or without prior exposure to hydrogen peroxide (H2O2). Samples were taken from the head kidney 4, 48, 125 and 192 h after immersion, and the regulation of several...

Assessment of Aquaflor (c), copper sulfate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female x Morone saxatilis male

Science.gov (United States)

Two experiments were conducted to assess different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass (SB) (Morone chrysops female x Morone saxatilis male). Experiment 1 assessed the efficacy of copper sulfate (CuSO4), florfenicol-medicated...

A thermostable serralysin inhibitor from marine bacterium Flavobacterium sp. YS-80-122

Science.gov (United States)

Liang, Pengjuan; Li, Shangyong; Wang, Kun; Wang, Fang; Xing, Mengxin; Hao, Jianhua; Sun, Mi

2018-03-01

Serralysin inhibitors have been proposed as potent drugs against many diseases and may help to prevent further development of antibiotic-resistant pathogenic bacteria. In this study, a novel serralysin inhibitor gene, lupI, was cloned from the marine bacterium Flavobacterium sp. YS-80-122 and expressed in Escherichia coli. The deduced serralysin inhibitor, LupI, shows <40% amino acid identity to other reported serralysin inhibitors. Multiple sequence alignment and phylogenetic analysis of LupI with other serralysin inhibitors indicated that LupI was a novel type of serralysin inhibitor. The inhibitory constant for LupI towards its target metalloprotease was 0.64 μmol/L. LupI was thermostable at high temperature, in which 35.6%-90.7% of its inhibitory activity was recovered after treatment at 100°C for 1-60 min followed by incubation at 0°C. This novel inhibitor may represent a candidate drug for the treatment of serralysin-related infections.

Application of a complete factorial design for the production of zeaxanthin by Flavobacterium sp.

Science.gov (United States)

Masetto, A; Flores-Cotera, L B; Díaz, C; Langley, E; Sanchez, S

2001-01-01

Utilizing a four-liter fermentor and applying a complete factorial design 2(3), the combined effects of agitation speed, aeration rate, and corn steep liquor concentration on zeaxanthin production by Flavobacterium sp. were studied. Maximum growth and production of total carotenoids and zeaxanthin were obtained at 600 rpm, 2 vvm and 4.6% corn steep liquor. Under these conditions, zeaxanthin represented 86% of the total carotenoids produced. Lower values of the variables studied resulted in lower growth, volumetric production of zeaxanthin and total carotenoids, and favored the formation of other carotenoids such as beta-carotene and canthaxanthin. The positive effects on growth and total carotenoids and zeaxanthin formation were in a large extent due to the interaction of agitation/corn steep liquor. However, aeration also had a positive effect on growth.

Proteomic analysis of the fish pathogen Flavobacterium columnare

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Lawrence Mark L

2010-06-01

Full Text Available Abstract Background Flavobacterium columnare causes columnaris disease in cultured and wild fish populations worldwide. Columnaris is the second most prevalent bacterial disease of commercial channel catfish industry in the United States. Despite its economic importance, little is known about the expressed proteins and virulence mechanisms of F. columnare. Here, we report the first high throughput proteomic analysis of F. columnare using 2-D LC ESI MS/MS and 2-DE MALDI TOF/TOF MS. Results Proteins identified in this study and predicted from the draft F. columnare genome were clustered into functional groups using clusters of orthologous groups (COGs, and their subcellular locations were predicted. Possible functional relations among the identified proteins were determined using pathway analysis. The total number of unique F. columnare proteins identified using both 2-D LC and 2-DE approaches was 621, of which 10.95% (68 were identified by both methods, while 77.29% (480 and 11.76% (73 were unique in 2-D LC and 2-DE, respectively. COG groupings and subcellular localizations were similar between our data set and proteins predicted from the whole genome. Twenty eight pathways were significantly represented in our dataset (P Conclusion Results from this study provide experimental evidence for many proteins that were predicted from the F. columnare genome annotation, and they should accelerate functional and comparative studies aimed at understanding virulence mechanisms of this important pathogen.

Augmentation of the antibody response of Atlantic salmon by oral administration of alginate-encapsulated IPNV antigens.

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Lihan Chen

Full Text Available The objective of the present study was to assess the effect of alginate-encapsulated infectious pancreatic necrosis virus antigens in inducing the immune response of Atlantic salmon as booster vaccines. One year after intraperitoneal injection with an oil-adjuvanted vaccine, post-smolts were orally boosted either by 1 alginate-encapsulated IPNV antigens (ENCAP; 2 soluble antigens (UNENCAP or 3 untreated feed (control. This was done twice, seven weeks apart. Sampling was done twice, firstly at 7 weeks post 1st oral boost and the 2nd, at 4 weeks after the 2nd oral boost. Samples included serum, head kidney, spleen and hindgut. Serum antibodies were analyzed by ELISA while tissues were used to assess the expression of IgM, IgT, CD4, GATA3, FOXP3, TGF-β and IL-10 genes by quantitative PCR. Compared to controls, fish fed with ENCAP had a significant increase (p<0.04 in serum antibodies following the 1st boost but not after the 2nd boost. This coincided with significant up-regulation of CD4 and GATA3 genes. In contrast, serum antibodies in the UNENCAP group decreased both after the 1st and 2nd oral boosts. This was associated with significant up-regulation of FOXP3, TGF-β and IL-10 genes. The expression of IgT was not induced in the hindgut after the 1st oral boost but was significantly up-regulated following the 2nd one. CD4 and GATA3 mRNA expressions exhibited a similar pattern to IgT in the hindgut. IgM mRNA expression on the other hand was not differentially regulated at any of the times examined. Our findings suggest that 1 Parenteral prime with oil-adjuvanted vaccines followed by oral boost with ENCAP results in augmentation of the systemic immune response; 2 Symmetrical prime and boost (mucosal with ENCAP results in augmentation of mucosal immune response and 3 Symmetrical priming and boosting (mucosal with soluble antigens results in the induction of systemic immune tolerance.

Efficiency of oxytetracycline treatment in rainbow trout experimentally infected with Flavobacterium psychrophilum strains having different in vitro antibiotic susceptibilities

DEFF Research Database (Denmark)

Bruun, Morten Sichlau; Madsen, Lone; Dalsgaard, Inger

2003-01-01

The medication effect of oxytetracycline on groups of rainbow trout fry experimentally infected with three strains of Flavobacterium psychrophilum was investigated. The infection model was based on intraperitoneal injection of the pathogen and treatment was done using medicated feed resulting...... in 100 mg oxytetracycline/kg fish for 10 days. The three F. psychrophilum strains had different antimicrobial susceptibilities and successful treatment was only obtained in the trial using a strain with a MICOTC of 0.25 mug/ml. No effect of treatment was seen in the group infected with a strain having...

Radiological examination of the spinal column in farmed rainbow trout Oncorhynchus mykiss (Walbaum): experiments with Flavobacterium psychrophilum and oxytetracycline

DEFF Research Database (Denmark)

Madsen, Lone; Arnbjerg, J.; Dalsgaard, Inger

2001-01-01

Flavobacterium psychrophilum and oxytetracycline have both been associated with spinal deformities in salmonids. Experiments were carried out to investigate whether infection with F, psychrophilum or medication with oxytetracycline (OTC) at the fry stage would result in an increased occurrence...... of infected fish. OTC treatments of up to 200 mg of OTC(kgfish)(-1) day(-1) for 10 days and repeated three times did not result in increased spinal deformities relative to untreated control groups; therefore, medication of rainbow trout with oxytetracycline did not cause deformities of the spinal column under...

Diversity of Flavobacterium psychrophilum and the potential use of its phages for protection against bacterial cold water disease in salmonids

DEFF Research Database (Denmark)

Castillo, D.; Higuera, G.; Villa, M.

2012-01-01

Flavobacterium psychrophilum causes rainbow trout fry syndrome (RTFS) and cold water disease (CWD) in salmonid aquaculture. We report characterization of F. psychrophilum strains and their bacteriophages isolated in Chilean salmonid aquaculture. Results suggest that under laboratory conditions ph...... together with the bacteria in a ratio of 10 plaque‐forming units per colony‐forming unit. While we recognize the artificial laboratory conditions used for these protection assays, this work is the first to demonstrate that phages might be able protect salmonids from RTFS or CWD....

Differential virulence mechanisms of infectious hematopoietic necrosis virus in rainbow trout (Oncorhynchus mykiss) include host entry and virus replication kinetics

Science.gov (United States)

Penaranda, M.M.D.; Purcell, M.K.; Kurath, G.

2009-01-01

Host specificity is a phenomenon exhibited by all viruses. For the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV), differential specificity of virus strains from the U and M genogroups has been established both in the field and in experimental challenges. In rainbow trout (Oncorhynchus mykiss), M IHNV strains are consistently more prevalent and more virulent than U IHNV. The basis of the differential ability of these two IHNV genogroups to cause disease in rainbow trout was investigated in live infection challenges with representative U and M IHNV strains. When IHNV was delivered by intraperitoneal injection, the mortality caused by U IHNV increased, indicating that the low virulence of U IHNV is partly due to inefficiency in entering the trout host. Analyses of in vivo replication showed that U IHNV consistently had lower prevalence and lower viral load than M IHNV during the course of infection. In analyses of the host immune response, M IHNV-infected fish consistently had higher and longer expression of innate immune-related genes such as Mx-1. This suggests that the higher virulence of M IHNV is not due to suppression of the immune response in rainbow trout. Taken together, the results support a kinetics hypothesis wherein faster replication enables M IHNV to rapidly achieve a threshold level of virus necessary to override the strong host innate immune response. ?? 2009 SGM.

Colorimetric Method of Loop-Mediated Isothermal Amplification with the Pre-Addition of Calcein for Detecting Flavobacterium columnare and its Assessment in Tilapia Farms.

Science.gov (United States)

Suebsing, Rungkarn; Kampeera, Jantana; Sirithammajak, Sarawut; Withyachumnarnkul, Boonsirm; Turner, Warren; Kiatpathomchai, Wansika

2015-03-01

Flavobacterium columnare, the causative agent of columnaris disease in fish, affects many economically important freshwater fish species. A colorimetric method of loop-mediated isothermal amplification with the pre-addition of calcein (LAMP-calcein) was developed and used to detect the presence of F. columnare in farmed tilapia (Nile Tilapia Oreochromis niloticus and red tilapia [Nile Tilapia × Mozambique Tilapia O. mossambicus]) and rearing water. The detection method, based on a change in color from orange to green, could be performed within 45 min at 63°C. The method was highly specific, as it had no cross-detections with 14 other bacterial species, including other fish pathogens and two Flavobacterium species. The method has a minimum detection limit of 2.2 × 10(2) F. columnare CFU; thus, it is about 10 times more sensitive than conventional PCR. With this method, F. columnare was detected in gonad, gill, and blood samples from apparently healthy tilapia broodstock as well as in samples of fertilized eggs, newly hatched fry, and rearing water. The bacteria isolated from the blood were further characterized biochemically and found to be phenotypically identical to F. columnare. The amplified products from the LAMP-calcein method had 97% homology with the DNA sequence of F. columnare.

Effect of temperature on the expression of IFN-1 (α, STAT-1 and Mx-1 genes in Oncorhynchus mykiss (Salmoniformes: Salmonidae exposed with the virus of the infectious pancreatic necrosis (IPNV

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Donald Arguedas Cortés

2015-06-01

Full Text Available The infectious pancreatic necrosis (IPNV is the causative agent of an acute illness well characterized in salmonids worldwide. Clinical signs and mortality rates are dependent on several factors such as the viral dose, the age of the fish, the water temperature, among others. An experimental study was conducted to measure the effect of temperature on the gene expression profile of IFN-1(α, STAT-1 and Mx-1 in rainbow trout fry, exposed to IPNV. Fry (n=198 were exposed at 8, 12 and 16°C, and samples were taken for 21 days to determine the virus titer and gene expression. In the first 11 days the greatest viral titer was recorded at 8°C compared with the values obtained at 12 and 16°C. At 8°C, there was a significant increase on day 4 of mRNA Mx-1 (t-test, p<0.05, time in which the viral titer began to decrease. Furthermore, as the viral titer increased, STAT-1 and Mx-1 (r=0.91 and (r=0.96 increased, respectively. The animals were able to recover from day 4 from some of the symptoms of IPN. Clinical disease was developed only in fish exposed to 12°C and all died between days 6 and 14, despite the highly significant increase shown in the average expression level of Mx-1, compared with the values recorded at 8°C and 16°C (Tukey, p<0.0001. Additionally, the expression profiles of IFN-1(α and STAT-1 decreased completely (~0.016 and (~0.020 times on day 7. The highest expression level of IFN-1(α, occurred at 16°C (Tukey, p<0.0005. Fry exposed at 16°C were normal during the experiment. IFN-1(α possibly generated a protector effect from day 2 when they showed a significant expression increase compared with the results at 8°C and 12°C (t-student, p<0.0001; however, STAT-1 was not significantly affected by temperature, although the highest average expression value was recorded at 16°C. Our research supports the expression of relevant anti-viral response genes as IFN-1(α, STAT-1 and Mx-1 are physiologically modulated by the water temperature

Efficacy of florfenicol, copper sulfate and potassium permanganate in controlling a natural infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass, Morone chrysops female x Morone saxatilis male

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Sunshine bass (Morone chrysops female ' Morone saxatilis male) naturally infected with Aeromonas hydrophila and Flavobacterium columnare were randomly assigned to six treatments: 1) two treatments of waterborne exposures to copper sulfate (CuSO4), at 2.1 and at 4.2 mg/L (approximately one and two pe...

Walleye Autochthonous Bacteria as Promising Probiotic Candidates against Flavobacterium columnare

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Hamza Seghouani

2017-07-01

Full Text Available Walleye (Sander vitreus is the second most fished freshwater species in Canada. While much sought by anglers, walleye also supports substantial commercial fisheries. To cope with the recent decline of wild walleye populations, fish farmers produce juveniles for lake stocking. However, walleye breeding is particularly tedious, mostly due to high disease susceptibility at larval and juvenile developmental stages. The main threat is the columnaris disease, which is caused by Flavobacterium columnare, an opportunistic bacteria. As F. columnare strains exhibit increasing antibiotic resistance, there is a strong need to develop efficient and sustainable alternative strategies to control columnaris disease. Bacterial probiotics have been shown to mitigate infections either by enhancing host immune response or by inhibiting pathogen growth. Being successfully assessed in many fish/pathogen combinations, we developed a tailored probiotic strategy for walleye to prevent and treat columnaris disease. Thirty-seven endogenous bacterial strains were isolated from healthy walleye’s skin and gut, were tested in vitro against F. columnare. Significant antagonistic effect against F. columnare was measured for 2 out of 37 endogenous strains. These two probiotic strains were identified as Pseudomonas fluorescens. The antagonistic effect of these two successful probiotics was further validated in vivo during a 2-month stress trial: groups receiving probiotic treatments showed on average 53.74% survival improvement.

Preparation and purification of Flavobacterium heparinum chondroitinases AC and B by hydrophobic interaction chromatography

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Aguiar J.A.K.

1999-01-01

Full Text Available Flavobacterium heparinum is a soil bacterium that produces several mucopolysaccharidases such as heparinase, heparitinases I and II, and chondroitinases AC, B, C and ABC. The purpose of the present study was to optimize the preparation of F. heparinum chondroitinases, which are very useful tools for the identification and structural characterization of chondroitin and dermatan sulfates. We observed that during the routine procedure for cell disruption (ultrasound, 100 kHz, 5 min some of the chondroitinase B activity was lost. Using milder conditions (2 min, most of the chondroitinase B and AC protein was solubilized and the enzyme activities were preserved. Tryptic soy broth without glucose was the best culture medium both for bacterial growth and enzyme induction. Chondroitinases AC and B were separated from each other and also from glucuronidases and sulfatases by hydrophobic interaction chromatography on HP Phenyl-Sepharose. A rapid method for screening of the column fractions was also developed based on the metachromatic shift of the color of dimethylmethylene blue.

Iron Overload Is Associated With Oxidative Stress and Nutritional Immunity During Viral Infection in Fish.

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Tarifeño-Saldivia, Estefanía; Aguilar, Andrea; Contreras, David; Mercado, Luis; Morales-Lange, Byron; Márquez, Katherine; Henríquez, Adolfo; Riquelme-Vidal, Camila; Boltana, Sebastian

2018-01-01

Iron is a trace element, essential to support life due to its inherent ability to exchange electrons with a variety of molecules. The use of iron as a cofactor in basic metabolic pathways is essential to both pathogenic microorganisms and their hosts. During evolution, the shared requirement of micro- and macro-organisms for this important nutrient has shaped the pathogen-host relationship. Infectious pancreatic necrosis virus (IPNv) affects salmonids constituting a sanitary problem for this industry as it has an important impact on post-smolt survival. While immune modulation induced by IPNv infection has been widely characterized on Salmo salar , viral impact on iron host metabolism has not yet been elucidated. In the present work, we evaluate short-term effect of IPNv on several infected tissues from Salmo salar . We observed that IPNv displayed high tropism to headkidney, which directly correlates with a rise in oxidative stress and antiviral responses. Transcriptional profiling on headkidney showed a massive modulation of gene expression, from which biological pathways involved with iron metabolism were remarkable. Our findings suggest that IPNv infection increase oxidative stress on headkidney as a consequence of iron overload induced by a massive upregulation of genes involved in iron metabolism.

Adaptive response to starvation in the fish pathogen Flavobacterium columnare: cell viability and ultrastructural changes

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Arias Covadonga R

2012-11-01

Full Text Available Abstract Background The ecology of columnaris disease, caused by Flavobacterium columnare, is poorly understood despite the economic losses that this disease inflicts on aquaculture farms worldwide. Currently, the natural reservoir for this pathogen is unknown but limited data have shown its ability to survive in water for extended periods of time. The objective of this study was to describe the ultrastructural changes that F. columnare cells undergo under starvation conditions. Four genetically distinct strains of this pathogen were monitored for 14 days in media without nutrients. Culturability and cell viability was assessed throughout the study. In addition, cell morphology and ultrastructure was analyzed using light microscopy, scanning electron microscopy, and transmission electron microscopy. Revival of starved cells under different nutrient conditions and the virulence potential of the starved cells were also investigated. Results Starvation induced unique and consistent morphological changes in all strains studied. Cells maintained their length and did not transition into a shortened, coccus shape as observed in many other Gram negative bacteria. Flavobacterium columnare cells modified their shape by morphing into coiled forms that comprised more than 80% of all the cells after 2 weeks of starvation. Coiled cells remained culturable as determined by using a dilution to extinction strategy. Statistically significant differences in cell viability were found between strains although all were able to survive in absence of nutrients for at least 14 days. In later stages of starvation, an extracellular matrix was observed covering the coiled cells. A difference in growth curves between fresh and starved cultures was evident when cultures were 3-months old but not when cultures were starved for only 1 month. Revival of starved cultures under different nutrients revealed that cells return back to their original elongated rod shape upon

Genomic Characterization of Flavobacterium psychrophilum Serotypes and Development of a Multiplex PCR-Based Serotyping Scheme

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Tatiana Rochat

2017-09-01

Full Text Available Flavobacterium psychrophilum is a devastating bacterial pathogen of salmonids reared in freshwater worldwide. So far, serological diversity between isolates has been described but the underlying molecular factors remain unknown. By combining complete genome sequence analysis and the serotyping method proposed by Lorenzen and Olesen (1997 for a set of 34 strains, we identified key molecular determinants of the serotypes. This knowledge allowed us to develop a robust multiplex PCR-based serotyping scheme, which was applied to 244 bacterial isolates. The results revealed a striking association between PCR-serotype and fish host species and illustrate the use of this approach as a simple and cost-effective method for the determination of F. psychrophilum serogroups. PCR-based serotyping could be a useful tool in a range of applications such as disease surveillance, selection of salmonids for bacterial coldwater disease resistance and future vaccine formulation.

Emergence of a new rhabdovirus associated with mass mortalities in eelpout (Zoarces viviparous) in the Baltic Sea.

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Axén, C; Hakhverdyan, M; Boutrup, T S; Blomkvist, E; Ljunghager, F; Alfjorden, A; Hagström, Å; Olesen, N J; Juremalm, M; Leijon, M; Valarcher, J-F

2017-02-01

We report the first description of a new Rhabdoviridae tentatively named eelpout rhabdovirus (EpRV genus Perhabdovirus). This virus was associated with mass mortalities in eelpout (Zoarces viviparous, Linnaeus) along the Swedish Baltic Sea coast line in 2014. Diseased fish showed signs of central nervous system infection, and brain lesions were confirmed by histology. A cytopathogenic effect was observed in cell culture, but ELISAs for the epizootic piscine viral haemorrhagic septicaemia virus (VHSV), infectious pancreas necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and spring viraemia of carp virus (SVCV) were negative. Further investigations by chloroform inactivation, indirect fluorescence antibody test and electron microscopy indicated the presence of a rhabdovirus. By deep sequencing of original tissue suspension and infected cell culture supernatant, the full viral genome was assembled and we confirmed the presence of a rhabdovirus with 59.5% nucleotide similarity to the closest relative Siniperca chuatsi rhabdovirus. The full-genome sequence of this new virus, eelpout rhabdovirus (EpRV), has been deposited in GenBank under accession number KR612230. An RT-PCR based on the L-gene sequence confirmed the presence of EpRV in sick/dead eelpout, but the virus was not found in control fish. Additional investigations to characterize the pathogenicity of EpRV are planned. © 2016 John Wiley & Sons Ltd.

Virulence of a chimeric recombinant infectious haematopoietic necrosis virus expressing the spring viraemia of carp virus glycoprotein in salmonid and cyprinid fish

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Emmenegger, Eveline; Biacchesi, Stéphane; Mérour, Emilie; Glenn, Jolene. A; Palmer, Alexander D.; Brémont, Michel; Kurath, Gael

2018-01-01

Infectious haematopoietic necrosis virus (IHNV) and spring viraemia of carp virus (SVCV) are both rhabdoviruses of fish, listed as notifiable disease agents by the World Organization for Animal Health. Recombinant rhabdoviruses with heterologous gene substitutions have been engineered to study genetic determinants and assess the potential of these recombinant viruses for vaccine development. A recombinant IHNV (rIHNV), containing the full-length genome of a European IHNV strain, was modified by deleting the glycoprotein (G) gene and replacing it with a European SVCV G-gene to make the rIHNV-Gsvcv. The chimeric rIHNV-Gsvcv level of virulence in rainbow trout, common carp and koi was assessed, and its ability to induce a protective immune response in surviving koi against wild-type SVCV infection was tested. The rIHNV-Gsvcv infection of trout led to high mortality, ranging from 78% to 92.5%, after immersion. In contrast, no deaths occurred in juvenile common carp after infection with rIHNV-Gsvcv by either immersion or intraperitoneal (IP) injection. Similarly, koi infected with rIHNV-Gsvcv via IP injection had little to no mortality (≤9%). Koi that survived initial infection with a high dose of recombinant virus rIHNV-Gsvcv were protected against a virulent SVCV challenge resulting in a high relative per cent survival of 82.5%.

Virulence of Flavobacterium columnare genomovars in rainbow trout Oncorhynchus mykiss.

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Evenhuis, Jason P; LaFrentz, Benjamin R

2016-08-09

Flavobacterium columnare is the causative agent of columnaris disease and is responsible for significant economic losses in aquaculture. F. columnare is a Gram-negative bacterium, and 5 genetic types or genomovars have been described based on restriction fragment length polymorphism of the 16S rRNA gene. Previous research has suggested that genomovar II isolates are more virulent than genomovar I isolates to multiple species of fish, including rainbow trout Oncorhynchus mykiss. In addition, improved genotyping methods have shown that some isolates previously classified as genomovar I, and used in challenge experiments, were in fact genomovar III. Our objective was to confirm previous results with respect to genomovar II virulence, and to determine the susceptibility of rainbow trout to other genomovars. The virulence of 8 genomovar I, 4 genomovar II, 3 genomovar II-B, and 5 genomovar III isolates originating from various sources was determined through 3 independent challenges in rainbow trout using an immersion challenge model. Mean cumulative percent mortality (CPM) of ~49% for genomovar I isolates, ~1% for genomovar II, ~5% for the II-B isolates, and ~7% for the III isolates was observed. The inability of genomovar II isolates to produce mortalities in rainbow trout was unanticipated based on previous studies, but may be due to a number of factors including rainbow trout source and water chemistry. The source of fish and/or the presence of sub-optimal environment may influence the susceptibility of rainbow trout to different F. columnare genomovars.

Atlantic salmon endothelial cells from the heart were more susceptible than fibroblasts from the bulbus arteriosus to four RNA viruses but protected from two viruses by dsRNA pretreatment.

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Pham, Phuc H; Tong, Winnie W L; Misk, Ehab; Jones, Ginny; Lumsden, John S; Bols, Niels C

2017-11-01

Heart diseases caused by viruses are major causes of Atlantic salmon aquaculture loss. Two Atlantic salmon cardiovascular cell lines, an endothelial cell line (ASHe) from the heart and a fibroblast cell line (BAASf) from the bulbus arteriosus, were evaluated for their response to four fish viruses, CSV, IPNV, VHSV IVa and VHSV IVb, and the innate immune agonist, double-stranded RNA mimic poly IC. All four viruses caused cytopathic effects in ASHe and BAASf. However, ASHe was more susceptible to all four viruses than BAASf. When comparing between the viruses, ASHe cells were found to be moderately susceptible to CSV and VHSV IVb, but highly susceptible to IPNV and VHSV IVa induced cell death. All four viruses were capable of propagating in the ASHe cell line, leading to increases in virus titre over time. In BAASf, CSV and IPNV produced more than one log increase in titre from initial infection, but VHSV IVb and IVa did not. When looking at the antiviral response of both cell lines, Mx proteins were induced in ASHe and BAASf by poly IC. All four viruses induced Mx proteins in BAASf, while only CSV and VHSV IVb induced Mx proteins in ASHe. IPNV and VHSV IVa suppressed Mx proteins expression in ASHe. Pretreatment of ASHe with poly IC to allow for Mx proteins accumulation protected the culture from subsequent infections with IPNV and VHSV IVa, resulting in delayed cell death, reduced virus titres and reduced viral proteins expression. These data suggest that endothelial cells potentially can serve as points of infections for viruses in the heart and that two of the four viruses, IPNV and VHSV IVa, have mechanisms to avoid or downregulate antiviral responses in ASHe cells. Furthermore, the high susceptibility of the ASHe cell line to IPNV and VHSV IVa can make it a useful tool for studying antiviral compounds against these viruses and for general detection of fish viruses. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characterization of isolates of Flavobacterium psychrophilum associated with coldwater disease or rainbow trout fry syndrome II: serological studies

DEFF Research Database (Denmark)

Lorenzen, Ellen; Olesen, Niels Jørgen

1997-01-01

The possibility of seroIogical differentiation between isolates of Flavobacterium psychrophilum was analyzed by ELISA and slide agglutination. Twenty-five Danish isolates and 20 isolates from other European countries were studied using polyclonal rabbit antisera and whole-cell preparations....... Unabsorbed as well as reciprocally absorbed antisera and purified Ig preparations derived from the antisera were included. Most of the isolates originated from clinical outbreaks of rainbow trout fry syndrome (RTFS) or coldwater disease (CWD), but some were isolated from asymptomatic fish or from other fish...... species with different disease signs, The ELISA showed the existence of different serotypes most distinctly, but slide agglutination supported the ELISA results. Three serotypes were found among the isolates studied: 1 major serotype (serotype Th) represented most of the Danish isolates and isolates from...

Development and characterization of two cell lines from gills of Atlantic salmon

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Gjessing, Mona C.; Aamelfot, Maria; Batts, William N.; Benestad, Sylvie L.; Dale, Ole B.; Thoen, Even; Weli, Simon C.; Winton, James R.

2018-01-01

Gill disease in Atlantic salmon, Salmo salar L., causes big losses in the salmon farming industry. Until now, tools to cultivate microorganisms causing gill disease and models to study the gill responses have been lacking. Here we describe the establishment and characterization of two cell lines from the gills of Atlantic salmon. Atlantic salmon gill cell ASG-10 consisted of cells staining for cytokeratin and e-cadherin and with desmosomes as seen by transmission electron microscopy suggesting the cells to be of epithelial origin. These structures were not seen in ASG-13. The cell lines have been maintained for almost 30 passages and both cell lines are fully susceptible to infection by infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus (VHSV), infectious pancreatic necrosis virus (IPNV), Atlantic salmon reovirus TS (TSRV) and Pacific salmon paramyxovirus (PSPV). While infectious salmon anemia virus (ISAV) did not cause visible CPE, immunofluorescent staining revealed a sub-fraction of cells in both the ASG-10 and ASG-13 lines may be permissive to infection. ASG-10 is able to proliferate and migrate to close scratches in the monolayer within seven days in vitro contrary to ASG-13, which does not appear to do have the same proliferative and migratory ability. These cell lines will be useful in studies of gill diseases in Atlantic salmon and may represent an important contribution for alternatives to experimental animals and studies of epithelial–mesenchymal cell biology.

Inter-laboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction methods used for detection of infectious pancreatic necrosis virus in Chile

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David Tapia

2017-07-01

Conclusions: Overall, the ring trial showed high values of sensitivity and specificity, with some problems of repeatability and inter-laboratory variability. This last issue needs to be addressed in order to allow harmonized diagnostic of IPNV within the country. We recommend the use of the NRL methods as validated and reliable qRT-PCR protocols for the detection of IPNV.

Larval and juvenile Pacific herring Clupea pallasii are not susceptible to infectious hematopoietic necrosis under laboratory conditions

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Hart, L.M.; Traxler, G.S.; Garver, K.A.; Richard, J.; Gregg, J.L.; Grady, C.A.; Kurath, G.; Hershberger, P.K.

2011-01-01

Infectious hematopoietic necrosis (IHN) leads to periodic epidemics among certain wild and farmed fish species of the Northeast (NE) Pacific. The source of the IHN virus (IHNV) that initiates these outbreaks remains unknown; however, a leading hypothesis involves viral persistence in marine host species such as Pacific herring Clupea pallasii. Under laboratory conditions we exposed specific pathogen-free (SPF) larval and juvenile Pacific herring to 103 to 104 plaque-forming units (pfu) of IHNV ml–1 by waterborne immersion. Cumulative mortalities among exposed groups were not significantly different from those of negative control groups. After waterborne exposure, IHNV was transiently recovered from the tissues of larvae but absent in tissues of juveniles. Additionally, no evidence of viral shedding was detected in the tank water containing exposed juveniles. After intraperitoneal (IP) injection of IHNV in juvenile herring with 103 pfu, IHNV was recovered from the tissues of sub-sampled individuals for only the first 5 d post-exposure. The lack of susceptibility to overt disease and transient levels of IHNV in the tissues of exposed fish indicate that Pacific herring do not likely serve a major epizootiological role in perpetuation of IHNV among free-ranging sockeye salmon Oncorhynchus nerka and farmed Atlantic salmon Salmo salar in the NE Pacific.

Evaluation of a Subunit Vaccine to Infectious Hematopoietic Necrosis Virus, July 31, 1989 to September 30, 1990, Annual Report.

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Leong, JoAnn Ching

1990-09-01

The IHNV glycoprotein has been identified as the virion protein which elicits neutralizing antibody in rabbits and induces protective immunity in fish to homologous and heterologous strains of IHNV (Engelking and Leong, 1989). These findings suggested that genetic engineering might be used to develop an economically feasible IHNV vaccine for fish. Thus, a clone of the IHNV glycoprotein gene was made and expression of a portion of this gene in bacteria resulted in a prototype IHNV subunit vaccine. Only 350 bases of IHNV sequence was expressed in this initial vaccine construction because there were 16 cysteine residues in the glycoprotein gene. Previous work with the rabies glycoprotein had shown that when the entire gene was expressed in bacteria, a denatured protein was produced, presumably because appropriate folding mechanisms for disulfide bond formation in protein were absent in E. coli. The IHNV vaccine clone contained a region of the gene which encoded only one cysteine residue. Despite the efficacy of the vaccine in laboratory trials, it seemed useful to determine whether other regions of the IHNV glycoprotein gene would be expressed in an antigenically recognizable form in bacteria and thereby, provide increased protection in fish. The recombinant plasmids pXL2, pXL3, and pXL7 were constructed so that all regions of the glycoprotein gene were expressed in bacteria as trpE-G fusion proteins. All of these recombinant plasmids produced fusion proteins that were also analyzed in Western immunoblots with anti-IHNV sera and specific monoclonal antibodies. These results were compared with the proteins produced by p52G and p618G, the plasmids identified in the original vaccine construction. The results of this comparison are shown.

Detection and quantification of Flavobacterium psychrophilum specific bacteriophages in rainbow trout upon different administration methods: Implications for disease control in aquaculture

DEFF Research Database (Denmark)

Christiansen, Rói Hammershaimb; Madsen, Lone; Dalsgaard, Inger

2013-01-01

Flavobacterium psychrophilum is the pathogen causing the disease rainbow trout fry syndrome (RTFS), which has important implications for aquaculture production and trade worldwide. RTFS can be treated by antibiotic administration, but with the increasing problem of antibiotic resistant bacteria...... in the internal organs in rainbow trout, following different administration methods. Three phage administration methods using phage FpV-9 were used: phage bath, oral administration of phagesuspension directly into the stomach and feeding with phage-coated feed pellets. Phages were detected in all the four...... examined organs (intestine, brain, spleen and liver) with all three administration methods, demonstrating that the phages are capable of passing the intestinal wall and entering the bloodstream. The highest phage concentration was found in the intestine where a maximum of 3x1010 phages g-1 was obtained...

Phylogenetic relationships of Iranian infectious hematopoietic necrosis virus of rainbow trout (Oncorhynchus mykiss) based on the glycoprotein gene

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Adel, Milad; Amiri, Alireza Babaalian; Dada, Maryam; Kurath, Gael; Laktarashi, Bahram; Ghajari, Amrolah; Breyta, Rachel

2016-01-01

Infectious hematopoietic necrosis virus (IHNV), a member of family Rhabdoviridae and genus Novirhabdoviridae, causes a highly lethal disease of salmon and trout. In Iran IHNV was first detected in 2001 on farms rearing rainbow trout (Oncorhynchus mykiss). To evaluate the genetic relationships of IHNV from northern and western Iran, the sequences of a 651-nt region of the glycoprotein gene were determined for two Iranian isolates. These sequences were analyzed to evaluate their genetic relatedness to worldwide isolates representing the five known genogroups of IHNV. Iranian isolates were most closely related to European isolates within the genogroup E rather than those of North American genogroups U, M and L, or the Asian genogroup J. It appears that Iranian IHNV was most likely introduced to Iran from a source in Europe by the movement of contaminated fish eggs.

Transcriptional responses of resistant and susceptible fish clones to the bacterial pathogen Flavobacterium psychrophilum.

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Christelle Langevin

Full Text Available Flavobacterium psychrophilum is a bacterial species that represents one of the most important pathogens for aquaculture worldwide, especially for salmonids. To gain insights into the genetic basis of the natural resistance to F. psychrophilum, we selected homozygous clones of rainbow trout with contrasted susceptibility to the infection. We compared the transcriptional response to the bacteria in the pronephros of a susceptible and a resistant line by micro-array analysis five days after infection. While the basal transcriptome of healthy fish was significantly different in the resistant and susceptible lines, the transcriptome modifications induced by the bacteria involved essentially the same genes and pathways. The response to F. psychrophilum involved antimicrobial peptides, complement, and a number of enzymes and chemokines. The matrix metalloproteases mmp9 and mmp13 were among the most highly induced genes in both genetic backgrounds. Key genes of both pro- and anti-inflammatory response such as IL1 and IL10, were up-regulated with a greater magnitude in susceptible animals where the bacterial load was also much higher. While higher resistance to F. psychrophilum does not seem to be based on extensive differences in the orientation of the immune response, several genes including complement C3 showed stronger induction in the resistant fish. They may be important for the variation of susceptibility to the infection.

Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase, FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01

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Akira Inoue

2015-01-01

Full Text Available In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11. The monosaccharide is non-enzymatically converted to 4-deoxy-l-ery thro-5-hexoseulose uronic acid (DEH, then reduced to 2-keto-3-deoxy-d-gluconate (KDG by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01.

Genetic and serological diversity of Flavobacterium psychrophilum isolates from salmonids in United Kingdom.

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Ngo, Thao P H; Bartie, Kerry L; Thompson, Kim D; Verner-Jeffreys, David W; Hoare, Rowena; Adams, Alexandra

2017-03-01

Flavobacterium psychrophilum is one of the most important bacterial pathogens affecting cultured rainbow trout (Oncorhynchus mykiss) and is increasingly causing problems in Atlantic salmon (Salmo salar L.) hatcheries. Little is known about the heterogeneity of F. psychrophilum isolates on UK salmonid farms. A total of 315 F. psychrophilum isolates, 293 of which were collected from 27 sites within the UK, were characterised using four genotyping methods and a serotyping scheme. A high strain diversity was identified among the isolates with 54 pulsotypes, ten (GTG) 5 -PCR types, two 16S rRNA allele lineages, seven plasmid profiles and three serotypes. Seven PFGE groups and 27 singletons were formed at a band similarity of 80%. PFGE group P (n=75) was found to be numerically predominant in eight sites within the UK. Two major PFGE clusters and 13 outliers were found at the band similarity of 40%. The predominant profileobserved within the F. psychrophilum isolates examined was PFGE cluster II - (GTG) 5 -PCR type r1-16S rRNA lineage II - serotype Th (70/156 isolates examined, 45%). Co-existence of genetically and serologically heterogeneous isolates within each farm was detected, confounding the ability to control RTFS outbreaks. The occurrence over time (up to 11 years) of F. psychrophilum pulsotypes in three representative sites (Scot I, Scot III and Scot V) within Scotland was examined, potentially providing important epidemiological data for farm management and the development of site-specific vaccines. Copyright © 2017 Elsevier B.V. All rights reserved.

Genomic Diversity and Evolution of the Fish Pathogen Flavobacterium psychrophilum

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Eric Duchaud

2018-02-01

Full Text Available Flavobacterium psychrophilum, the etiological agent of rainbow trout fry syndrome and bacterial cold-water disease in salmonid fish, is currently one of the main bacterial pathogens hampering the productivity of salmonid farming worldwide. In this study, the genomic diversity of the F. psychrophilum species is analyzed using a set of 41 genomes, including 30 newly sequenced isolates. These were selected on the basis of available MLST data with the two-fold objective of maximizing the coverage of the species diversity and of allowing a focus on the main clonal complex (CC-ST10 infecting farmed rainbow trout (Oncorhynchus mykiss worldwide. The results reveal a bacterial species harboring a limited genomic diversity both in terms of nucleotide diversity, with ~0.3% nucleotide divergence inside CDSs in pairwise genome comparisons, and in terms of gene repertoire, with the core genome accounting for ~80% of the genes in each genome. The pan-genome seems nevertheless “open” according to the scaling exponent of a power-law fitted on the rate of new gene discovery when genomes are added one-by-one. Recombination is a key component of the evolutionary process of the species as seen in the high level of apparent homoplasy in the core genome. Using a Hidden Markov Model to delineate recombination tracts in pairs of closely related genomes, the average recombination tract length was estimated to ~4.0 Kbp and the typical ratio of the contributions of recombination and mutations to nucleotide-level differentiation (r/m was estimated to ~13. Within CC-ST10, evolutionary distances computed on non-recombined regions and comparisons between 22 isolates sampled up to 27 years apart suggest a most recent common ancestor in the second half of the nineteenth century in North America with subsequent diversification and transmission of this clonal complex coinciding with the worldwide expansion of rainbow trout farming. With the goal to promote the development of

Replication of Infectious Pancreatic Necrosis Virus in Different Cell Lines and in Rainbow Trout (Oncorhynchus Mykiss Fingerlings

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Matvienko Natalija

2014-07-01

Full Text Available The results of a study of Infectious Pancreatic Necrosis Virus (IPNV isolated in natural reservoirs in Ukraine are presented. The pathogenicity of isolates was investigated in vitro on cell cultures and in vivo on rainbow trout, Oncorhynchus mykiss (Walbaum, fingerlings. Experimental indications were that the Ukrainian IPNV isolates have affinity with reference European strains. During the reproduction of these isolates in cell cultures of FHM (fat head minnow, RTG-2 (rainbow trout gonads, and BF-2 (bluegill caudal peduncle, complicated degenerative changes were visible that finally led to the full destruction of cell monolayers. The experimental infection of rainbow trout fingerlings resulted in typical disease symptoms that were systemic. However, obvious evidence of viral infection was noted in single individuals only, and the majority of experimental fish died without visible disease symptoms. During the study of physicochemical properties, it was noted that Ukrainian isolates completely lost their infectivity with chloroform treatment and heating to 60°C. This proved that IPNV isolates are resistant to Ion concentrations in the range of pH 3.0 to 12.0.

Complete genome sequence analysis of the fish pathogen Flavobacterium columnare provides insights into antibiotic resistance and pathogenicity related genes.

Science.gov (United States)

Zhang, Yulei; Zhao, Lijuan; Chen, Wenjie; Huang, Yunmao; Yang, Ling; Sarathbabu, V; Wu, Zaohe; Li, Jun; Nie, Pin; Lin, Li

2017-10-01

We analyzed here the complete genome sequences of a highly virulent Flavobacterium columnare Pf1 strain isolated in our laboratory. The complete genome consists of a 3,171,081 bp circular DNA with 2784 predicted protein-coding genes. Among these, 286 genes were predicted as antibiotic resistance genes, including 32 RND-type efflux pump related genes which were associated with the export of aminoglycosides, indicating inducible aminoglycosides resistances in F. columnare. On the other hand, 328 genes were predicted as pathogenicity related genes which could be classified as virulence factors, gliding motility proteins, adhesins, and many putative secreted proteases. These genes were probably involved in the colonization, invasion and destruction of fish tissues during the infection of F. columnare. Apparently, our obtained complete genome sequences provide the basis for the explanation of the interactions between the F. columnare and the infected fish. The predicted antibiotic resistance and pathogenicity related genes will shed a new light on the development of more efficient preventional strategies against the infection of F. columnare, which is a major worldwide fish pathogen. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of Four Distinct Phylogenetic Groups in Flavobacterium columnare With Fish Host Associations

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Benjamin R. LaFrentz

2018-03-01

Full Text Available Columnaris disease, caused by the Gram-negative bacterium Flavobacterium columnare, is one of the most prevalent fish diseases worldwide. An exceptionally high level of genetic diversity among isolates of F. columnare has long been recognized, whereby six established genomovars have been described to date. However, little has been done to quantify or characterize this diversity further in a systematic fashion. The objective of this research was to perform phylogenetic analyses of 16S rRNA and housekeeping gene sequences to decipher the genetic diversity of F. columnare. Fifty isolates and/or genomes of F. columnare, originating from diverse years, geographic locations, fish hosts, and representative of the six genomovars were analyzed in this study. A multilocus phylogenetic analysis (MLPA of the 16S rRNA and six housekeeping genes supported four distinct F. columnare genetic groups. There were associations between genomovar and genetic group, but these relationships were imperfect indicating that genomovar assignment does not accurately reflect F. columnare genetic diversity. To expand the dataset, an additional 90 16S rRNA gene sequences were retrieved from GenBank and a phylogenetic analysis of this larger dataset also supported the establishment of four genetic groups. Examination of isolate historical data indicated biological relevance to the identified genetic diversity, with some genetic groups isolated preferentially from specific fish species or families. It is proposed that F. columnare isolates be assigned to the four genetic groups defined in this study rather than genomovar in order to facilitate a standard nomenclature across the scientific community. An increased understanding of which genetic groups are most prevalent in different regions and/or aquaculture industries may allow for the development of improved targeted control and treatment measures for columnaris disease.

Susceptibility of Koi and Yellow Perch to infectious hematopoietic necrosis virus by experimental exposure

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Palmer, Alexander D.; Emmenegger, Eveline J.

2014-01-01

Infectious hematopoietic necrosis virus (IHNV) is a novirhabdoviral pathogen that originated in western North America among anadromous Pacific salmonids. Severe disease epidemics in the late 1970s resulting from IHNV's invasion into farmed Rainbow Trout Oncorhynchus mykiss in North America, Asia, and Europe emphasized IHNV's ability to adapt to new hosts under varying rearing conditions. Yellow Perch Perca flavescens and Koi Carp Cyprinus carpio (hereafter, “Koi”) are aquaculture-reared fish that are highly valued in sport fisheries and the ornamental fish trade, respectively, but it is unknown whether these fish species are vulnerable to IHNV infection. In this study, we exposed Yellow Perch, Koi, and steelhead (anadromous Rainbow Trout) to IHNV by intraperitoneal injection (106 PFU/fish) and by immersion (5.7×105 PFU/mL) for 7 h, and monitored fish for 28 d. The extended immersion exposure and high virus concentrations used in the challenges were to determine if the tested fish had any level of susceptibility. After experimental exposure, Yellow Perch and Koi experienced low mortality (35%). Virus was found in dead fish of all species tested and in surviving Yellow Perch by plaque assay and quantitative reverse transcription polymerase chain reaction (qPCR), with a higher prevalence in Yellow Perch than Koi. Infectious virus was also detected in Yellow Perch out to 5 d after bath challenge. These findings indicate that Yellow Perch and Koi are highly resistant to IHNV disease under the conditions tested, but Yellow Perch are susceptible to infection and may serve as possible virus carriers.

In vitro infection of salmonid epidermal tissues by infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus

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Yamamoto, T.; Batts, W.N.; Winton, J.R.

1992-01-01

The ability of two rhabdoviruses, infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV), to infect fish skin was investigated by in vitro infection of excised tissues. Virus replication was determined by plaque assay of homogenized tissue extracts, and the virus antigen was detected by immunohistology of tissue sections. Gill, fin, and ventral abdominal skin tissues of rainbow trout Oncorhynchus mykiss that had been infected in vitro with a virulent strain of IHNV (193–110) produced substantial increases in virus titer within 24 h. Titers continued to increase up until day 3 of incubation; by this time, virus had increased 1,000-fold or more. This increase in IHNV titer occurred in epidermal tissues of fingerlings and of older fish. In another experiment, IHNV replicated in excised rainbow trout tissues whether the fish had been subject to prior infection with a virulent strain of IHNV (Western Regional Aquaculture Consortium isolate) or whether the fish had been infected previously with an attenuated strain of the virus (Nan Scott Lake, with 100 passes in culture). A virulent strain of VHSV (23/75) replicated effectively in excised gill tissues and epidermal tissues of rainbow trout and chinook salmon O. tshawytscha; however, the avirulent North American strain of VHSV (Makah) replicated poorly or not at all.

Genomics and physiology of a marine flavobacterium encoding a proteorhodopsin and a xanthorhodopsin-like protein.

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Thomas Riedel

Full Text Available Proteorhodopsin (PR photoheterotrophy in the marine flavobacterium Dokdonia sp. PRO95 has previously been investigated, showing no growth stimulation in the light at intermediate carbon concentrations. Here we report the genome sequence of strain PRO95 and compare it to two other PR encoding Dokdonia genomes: that of strain 4H-3-7-5 which shows the most similar genome, and that of strain MED134 which grows better in the light under oligotrophic conditions. Our genome analysis revealed that the PRO95 genome as well as the 4H-3-7-5 genome encode a protein related to xanthorhodopsins. The genomic environment and phylogenetic distribution of this gene suggest that it may have frequently been recruited by lateral gene transfer. Expression analyses by RT-PCR and direct mRNA-sequencing showed that both rhodopsins and the complete β-carotene pathway necessary for retinal production are transcribed in PRO95. Proton translocation measurements showed enhanced proton pump activity in response to light, supporting that one or both rhodopsins are functional. Genomic information and carbon source respiration data were used to develop a defined cultivation medium for PRO95, but reproducible growth always required small amounts of yeast extract. Although PRO95 contains and expresses two rhodopsin genes, light did not stimulate its growth as determined by cell numbers in a nutrient poor seawater medium that mimics its natural environment, confirming previous experiments at intermediate carbon concentrations. Starvation or stress conditions might be needed to observe the physiological effect of light induced energy acquisition.

Characterization of structural proteins of hirame rhabdovirus, HRV

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Nishizawa, Toyohiko; Yoshimizu, Mamoru; Winton, James; Ahne, Winfried; Kimura, Takahisa

1991-01-01

Structural proteins of hirame rhabdovirus (HRV) were analyzed by SDS-polyacrylarnide gel electrophoresis, western blotting, 2-dimensional gel electrophoresis, and Triton X-100 treatment. Purified HRV virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N), and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 156 KDa; G, 68 KDa; N, 46.4 KDa; M1, 26.4 KDa; and M2, 19.9 KDa. The electrophorehc pattern formed by the structural proteins of HRV was clearly different from that formed by pike fry rhabdovirus, spring viremia of carp virus, eel virus of America, and eel virus European X which belong to the Vesiculovirus genus; however, it resembled the pattern formed by structural proteins of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) which are members of the Lyssavirus genus. Among HRV, IHNV, and VHSV, differences were observed in the relative mobilities of the G, N, M1, and M2 proteins. Western blot analysis revealed that the G. N, and M2 proteins of HRV shared antigenic determinants with IHNV and VHSV, but not with any of the 4 fish vesiculoviruses tested. Cross-reactions between the M1 proteins of HRV, IHNV, or VHSV were not detected in this assay. Two-dimensional gel electrophoresis was used to show that HRV differed from IHNV or VHSV in the isoelectric point (PI) of the M1 and M2 proteins. In this system, 2 forms of the M1 protein of HRV and IHNV were observed.These subspecies of M1 had the same relative mobility but different p1 values. Treatment of purified virions with 2% Triton X-100 in Tris buffer containing NaCl removed the G, M1, and M2 proteins of IHNV, but HRV virions were more stable under these conditions.

Flavobacterium psychrophilum y su patología en alevines de Onchorhynchus mykiss del centro piscícola El Ingenio, Huancayo

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Jorge León

2008-12-01

Full Text Available En el presente trabajo se reporta la presencia de Flavobacterium psychrophilum, como agente causante de la patología “enfermedad bacteriana del agua fría” en alevines de Onchorhynchus mykiss “trucha arco iris” del Centro Piscícola El Ingenio, Junín (3250 m de altitud. La lesión macroscópica externa más frecuente fue la ulceración profunda de la región dorsal del pez acompañado de un ennegrecimiento localizado de la piel. Internamente se observó una marcada esplenomegalia, palidez del hígado, riñón y branquias, inflamación del intestino y acumulación de líquido ascítico en el peritoneo. No se detectó hemorragia interna. En Agar Cytophaga Modificado (ACM según Anaker & Ordal (1959 fueron aisladas inicialmente 29 Gram negativas, de las cuales según la caracterización fenotípica y pruebas bioquímicas 9 fueron consideradas como F. psychrophilum. Pruebas de susceptibilidad antibiótica mostraron alta sensibilidad de las cepas a Gentamicina, Ceftazidina, oxitetraciclina, Norfloxacina, furazolidona, Ciprofloxacina y Cefoxitina.

Transmission routes maintaining a viral pathogen of steelhead trout within a complex multi-host assemblage

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Breyta, Rachel; Brito, Ilana L.; Ferguson, Paige; Kurath, Gael; Naish, Kerry A.; Purcell, Maureen; Wargo, Andrew R.; LaDeau, Shannon L.

2017-01-01

This is the first comprehensive region wide, spatially explicit epidemiologic analysis of surveillance data of the aquatic viral pathogen infectious hematopoietic necrosis virus (IHNV) infecting native salmonid fish. The pathogen has been documented in the freshwater ecosystem of the Pacific Northwest of North America since the 1950s, and the current report describes the disease ecology of IHNV during 2000–2012. Prevalence of IHNV infection in monitored salmonid host cohorts ranged from 8% to 30%, with the highest levels observed in juvenile steelhead trout. The spatial distribution of all IHNV-infected cohorts was concentrated in two sub-regions of the study area, where historic burden of the viral disease has been high. During the study period, prevalence levels fluctuated with a temporal peak in 2002. Virologic and genetic surveillance data were analyzed for evidence of three separate but not mutually exclusive transmission routes hypothesized to be maintaining IHNV in the freshwater ecosystem. Transmission between year classes of juvenile fish at individual sites (route 1) was supported at varying levels of certainty in 10%–55% of candidate cases, transmission between neighboring juvenile cohorts (route 2) was supported in 31%–78% of candidate cases, and transmission from adult fish returning to the same site as an infected juvenile cohort was supported in 26%–74% of candidate cases. The results of this study indicate that multiple specific transmission routes are acting to maintain IHNV in juvenile fish, providing concrete evidence that can be used to improve resource management. Furthermore, these results demonstrate that more sophisticated analysis of available spatio-temporal and genetic data is likely to yield greater insight in future studies.

Genetic variation underlying resistance to infectious hematopoietic necrosis virus in a steelhead trout (Oncorhynchus mykiss) population

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Brieuc, Marine S. O.; Purcell, Maureen K.; Palmer, Alexander D.; Naish, Kerry A.

2015-01-01

Understanding the mechanisms of host resistance to pathogens will allow insights into the response of wild populations to the emergence of new pathogens. Infectious hematopoietic necrosis virus (IHNV) is endemic to the Pacific Northwest and infectious to Pacific salmon and trout (Oncorhynchus spp.). Emergence of the M genogroup of IHNV in steelhead trout O. mykiss in the coastal streams of Washington State, between 2007 and 2011, was geographically heterogeneous. Differences in host resistance due to genetic change were hypothesized to be a factor influencing the IHNV emergence patterns. For example, juvenile steelhead trout losses at the Quinault National Fish Hatchery (QNFH) were much lower than those at a nearby facility that cultures a stock originally derived from the same source population. Using a classical quantitative genetic approach, we determined the potential for the QNFH steelhead trout population to respond to selection caused by the pathogen, by estimating the heritability for 2 traits indicative of IHNV resistance, mortality (h2 = 0.377 (0.226 - 0.550)) and days to death (h2 = 0.093 (0.018 - 0.203)). These results confirm that there is a genetic basis for resistance and that this population has the potential to adapt to IHNV. Additionally, genetic correlation between days to death and fish length suggests a correlated response in these traits to selection. Reduction of genetic variation, as well as the presence or absence of resistant alleles, could affect the ability of populations to adapt to the pathogen. Identification of the genetic basis for IHNV resistance could allow the assessment of the susceptibility of other steelhead populations.

Data on consistency among different methods to assess atherosclerotic plaque echogenicity on standard ultrasound and intraplaque neovascularization on contrast-enhanced ultrasound imaging in human carotid artery

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Mattia Cattaneo

2016-12-01

Full Text Available Here we provide the correlation among different carotid ultrasound (US variables to assess echogenicity n standard carotid US and to assess intraplaque neovascularization on contrast enhanced US. We recruited 45 consecutive subjects with an asymptomatic≥50% carotid artery stenosis. Carotid plaque echogenicity at standard US was visually graded according to Gray–Weale classification (GW and measured by the greyscale median (GSM, a semi-automated computerized measurement performed by Adobe Photoshop®. On CEUS imaging IPNV was graded according to the visual appearance of contrast within the plaque according to three different methods: CEUS_A (1=absent; 2=present; CEUS_B a three-point scale (increasing IPNV from 1 to 3; CEUS_C a four-point scale (increasing IPNV from 0 to 3. We have also implemented a new simple quantification method derived from region of interest (ROI signal intensity ratio as assessed by QLAB software. Further information is available in “Contrast-enhanced ultrasound imaging of intraplaque neovascularization and its correlation to plaque echogenicity in human carotid arteries atherosclerosis (M. Cattaneo, D. Staub, A.P. Porretta, J.M. Gallino, P. Santini, C. Limoni et al., 2016 [1].

Genetic diversity and antimicrobial resistance of Flavobacterium psychrophilum isolated from cultured rainbow trout, Onchorynchus mykiss (Walbaum), in Spain.

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Del Cerro, A; Márquez, I; Prieto, J M

2010-04-01

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease (CWD) and rainbow trout fry syndrome (RTFS) in salmonids. These diseases are a major problem in the aquaculture industry in Spain, and a better understanding of the epidemiology of F. psychrophilum isolates is necessary to improve management strategies. In this study, to investigate genetic variability of this bacterium, pulsed-field gel electrophoresis after DNA digestion with endonuclease StuI, plasmid profiling analysis and antimicrobial susceptibility testing were undertaken with 25 isolates of F. psychrophilum from Spain. These isolates were classified into 17 patterns by PFGE analysis, which were grouped into four clusters and seven independent branches. Twenty isolates (80%) possessed plasmids of 3.5 kb (n = 13) or 5.5 kb (n = 7). No plasmids were associated with antibiotic resistance to oxytetracycline (OTC) or florfenicol (FLO). Twenty isolates (80%) had minimum inhibitory concentrations (MICs) to OTC of between 2.4 and 9.7 microg mL(-1), and all isolates were susceptible to FLO. A relationship between the origin of the isolates and PFGE genotypes was found. Plasmid profile typing correlated with PFGE profile typing, whereas no correlation was found between antimicrobial susceptibility testing and PFGE profiles. These results suggest that the population of F. psychrophilum with pathogenic potential in northern Spain is quite heterogeneous.

Genomic makeup of the marine flavobacterium Nonlabens (Donghaeana) dokdonensis and identification of a novel class of rhodopsins.

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Kwon, Soon-Kyeong; Kim, Byung Kwon; Song, Ju Yeon; Kwak, Min-Jung; Lee, Choong Hoon; Yoon, Jung-Hoon; Oh, Tae Kwang; Kim, Jihyun F

2013-01-01

Rhodopsin-containing marine microbes such as those in the class Flavobacteriia play a pivotal role in the biogeochemical cycle of the euphotic zone (Fuhrman JA, Schwalbach MS, Stingl U. 2008. Proteorhodopsins: an array of physiological roles? Nat Rev Microbiol. 6:488-494). Deciphering the genome information of flavobacteria and accessing the diversity and ecological impact of microbial rhodopsins are important in understanding and preserving the global ecosystems. The genome sequence of the orange-pigmented marine flavobacterium Nonlabens dokdonensis (basonym: Donghaeana dokdonensis) DSW-6 was determined. As a marine photoheterotroph, DSW-6 has written in its genome physiological features that allow survival in the oligotrophic environments. The sequence analysis also uncovered a gene encoding an unexpected type of microbial rhodopsin containing a unique motif in addition to a proteorhodopsin gene and a number of photolyase or cryptochrome genes. Homologs of the novel rhodopsin gene were found in other flavobacteria, alphaproteobacteria, a species of Cytophagia, a deinococcus, and even a eukaryote diatom. They all contain the characteristic NQ motif and form a phylogenetically distinct group. Expression analysis of this rhodopsin gene in DSW-6 indicated that it is induced at high NaCl concentrations, as well as in the presence of light and the absence of nutrients. Genomic and metagenomic surveys demonstrate the diversity of the NQ rhodopsins in nature and the prevalent occurrence of the encoding genes among microbial communities inhabiting hypersaline niches, suggesting its involvement in sodium metabolism and the sodium-adapted lifestyle.

Alkaline active cyanide dihydratase of Flavobacterium indicum MTCC 6936: Growth optimization, purification, characterization and in silico analysis.

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Kumar, Virender; Kumar, Vijay; Bhalla, Tek Chand

2018-05-15

The present work explores a rare cyanide dihydratase of Flavobacterium indicum MTCC 6936 for its potential of cyanide degradation. The enzyme is purified to 12 fold with a yield of 76%. SDS and native-PAGE analysis revealed that enzyme was monomer of 40 kDa size. The enzyme works well in mesophilic range at wide array of pH. The thermostability profile of cyanide dihydratase revealed that the enzyme is quite stable at 30 °C and 35 °C with half-life of 6 h 30 min and 5 h respectively. K m and V max for cyanide dihydratase of F. indicum was measured to be 4.76 mM and 45 U mg -1 with k cat calculated to be 27.3 s -1 and specificity constant (k cat /K m ) to be around 5.67 mM -1  s -1 . MALDI-TOF analysis of purified protein revealed that the amino acid sequence has 50% and 43% sequence identity with putative amino acid sequence of F. indicum and earlier reported cyanide dihydratase of Bacillus pumilus respectively. Homology modeling studies of cyanide dihydratase of F. indicum predicted the catalytic triad of the enzyme indicating Cys at 164, Glu at 46 and Lys at 130th position. The purified enzyme has potential applications in bioremediation and analytical sector. Copyright © 2017. Published by Elsevier B.V.

Optimization of nested polymerase chain reaction assays for identification of Aeromonas salmonicida, Yersinia ruckeri and Flavobacterium psychrophilum

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Taylor, P.W.; Winton, J.R.

2002-01-01

Nested polymerase chain reaction (PCR) assays were developed using first-round primers complementary to highly conserved regions within the bacterial 16S ribosomal RNA (rRNA) gene (universal eubacterial primers) and second-round primers specific for sequences within the 16S rRNA genes of Aeromonas salmonicida, Yersinia ruckeri, andFlavobacterium psychrophilum. Following optimization of the MgCl2 concentration and primer annealing temperature, PCR employing the universal eubacterial primers was used to amplify a 1,500-base-pair (bp) product visible in agarose gels stained with ethidium bromide. The calculated detection limit of this single-round assay was less than 1.4 × 104 colony-forming units (CFU) per reaction for all bacterial species tested. Single-round PCR using primer sets specific for A. salmonicida, Y. ruckeri, and F. psychrophilumamplified bands of 271, 575, and 1,100 bp, respectively, with detection limits of less than 1.4 × 104, 1.4 × 105, and 1.4 × 105 CFU per reaction. Using the universal eubacterial primers in the first round and the species-specific primer sets in the second round of nested PCR assays improved the detection ability by approximately four orders of magnitude to fewer than 14 CFU per sample for each of the three bacterial species. Such nested assays could be adapted to a wide variety of bacterial fish pathogens for which 16S sequences are available.

High virulence differences among phylogenetically distinct isolates of the fish rhabdovirus viral hemorrhagic septicaemia virus are not explained by variability of the surface glycoprotein G or the non-virion protein Nv

DEFF Research Database (Denmark)

Einer-Jensen, Katja; Harmache, Abdallah; Biacchesi, Stéphane

2014-01-01

-related novirhabdovirus [infectious hematopoietic necrosis virus (IHNV)], four chimaeric IHNV–VHSV recombinant viruses were generated. These chimaeric viruses included substitution of the IHNV glyco- (G) or non-structural (Nv) protein with their counterparts from either a trout-derived or a marine VHSV strain....... Comparative challenge experiments in rainbow trout fingerlings revealed similar levels of survival induced by the recombinant (r)IHNV–VHSV chimaeric viruses regardless of whether the G or Nv genes originated from VHSV isolated from a marine fish species or from rainbow trout. Interestingly, recombinant IHNV...... gained higher virulence following substitution of the G gene with those of the VHSV strains, whilst the opposite was the case following substitution of the Nv genes....

Molecular epidemiology and evolution of fish Novirhabdoviruses

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Kurath, Gael

2014-01-01

The genus Novirhabdoviridae contains several of the important rhabdoviruses that infect fish hosts. There are four established virus species: Infectious hematopoietic necrosis virus (IHNV), Viral hemorrhagic septicemia virus (VHSV), Hirame rhabdovirus(HIRRV), and Snakehead rhabdovirus (SHRV). Viruses of these species vary in host and geographic range, and they have all been studied at the molecular and genomic level. As globally significant pathogens of cultured fish, IHNV and VHSV have been particularly well studied in terms of molecular epidemiology and evolution. Phylogenic analyses of hundreds of field isolates have defined five major genogroups of IHNV and four major genotypes of VHSV worldwide. These phylogenies are informed by the known histories of IHNV and VHSV, each involving a series of viral emergence events that are sometimes associated with host switches, most often into cultured rainbow trout. In general, IHNV has relatively low genetic diversity and a narrow host range, and has been spread from its endemic source in North American to Europe and Asia due to aquaculture activities. In contrast, VHSV has broad host range and high genetic diversity, and the source of emergence events is virus in widespread marine fish reservoirs in the northern Atlantic and Pacific Oceans. Common mechanisms of emergence and host switch events include use of raw feed, proximity to wild fish reservoirs of virus, and geographic translocations of virus or naive fish hosts associated with aquaculture.

Variable Number of Tandem Repeats (VNTR) analysis of Flavobacterium psychrophilum from salmonids in Chile and Norway.

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Apablaza, Patricia; Brevik, Øyvind J; Mjøs, Svein; Valdebenito, Samuel; Ilardi, Pedro; Battaglia, Juan; Dalsgaard, Inger; Nylund, Are

2015-07-14

Flavobacterium psychrophilum causes serious fish diseases such RTFS and BCWD, affecting the aquaculture industry worldwide. Commercial vaccines are not available and control of the disease depends on the use of antibiotics. Reliable methods for detection and identification of different isolates of this bacterium could play an important role in the development of good management strategies. The aim of this study was to identify genetic markers for discrimination between isolates. A selection of eight VNTRs from 53 F. psychrophilum isolates from Norway, Chile, Denmark and Scotland were analyzed. The results were compared with previous work on the same pathogen using MLST for genetic differentiation. The VNTR analysis gave a separation between the F. psychrophilum isolates supporting the results of previous MLST work. A higher diversity was found among the Chilean isolates compared to those from Norway, which suggests a more homogenous reservoir in Norway. Transgenerational transmission of F. psychrophilum from other countries, exporting salmon embryos to Chile, may explain the differences in diversity. The same transmission mechanisms could also explain the wide geographical distribution of identical isolates in Norway. But, this could also be a result of movement of smolts and embryos. The selected VNTRs are stable genetic markers and no variation was observed after several passages on agar plates at different temperatures. These VNTRs are important additions for genotyping of F. psychrophilum isolates. Future studies on VNTRs of F. psychrophilum should include isolates from more host species from a wider geographical area. To get a more robust genotyping the VNTRs should be used in concert with MLST. Future studies of isolates with high and low virulence should focus on identifying virulence markers using VTNRs and MLST.

Profilings of MicroRNAs in the Liver of Common Carp (Cyprinus carpio) Infected with Flavobacterium columnare

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Zhao, Lijuan; Lu, Hong; Meng, Qinglei; Wang, Jinfu; Wang, Weimin; Yang, Ling; Lin, Li

2016-01-01

MicroRNAs (miRNAs) play important roles in regulation of many biological processes in eukaryotes, including pathogen infection and host interactions. Flavobacterium columnare (FC) infection can cause great economic loss of common carp (Cyprinus carpio) which is one of the most important cultured fish in the world. However, miRNAs in response to FC infection in common carp has not been characterized. To identify specific miRNAs involved in common carp infected with FC, we performed microRNA sequencing using livers of common carp infected with and without FC. A total of 698 miRNAs were identified, including 142 which were identified and deposited in the miRbase database (Available online: http://www.mirbase.org/) and 556 had only predicted miRNAs. Among the deposited miRNAs, eight miRNAs were first identified in common carp. Thirty of the 698 miRNAs were differentially expressed miRNAs (DIE-miRNAs) between the FC infected and control samples. From the DIE-miRNAs, seven were selected randomly and their expression profiles were confirmed to be consistent with the microRNA sequencing results using RT-PCR and qRT-PCR. In addition, a total of 27,363 target genes of the 30 DIE-miRNAs were predicted. The target genes were enriched in five Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including focal adhesion, extracellular matrix (ECM)-receptor interaction, erythroblastic leukemia viral oncogene homolog (ErbB) signaling pathway, regulation of actin cytoskeleton, and adherent junction. The miRNA expression profile of the liver of common carp infected with FC will pave the way for the development of effective strategies to fight against FC infection. PMID:27092486

The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: Analysis of relationships with other rhabdoviruses

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Bjorklund, H.V.; Higman, K.H.; Kurath, G.

1996-01-01

The nucleotide sequences of the glycoprotein genes and all of the internal gene junctions of the fish pathogenic rhabdoviruses spring viremia of carp virus (SVCV) and hirame rhabdovirus (HIRRV) have been determined from cDNA clones generated from viral genomic RNA. The SVCV glycoprotein gene sequence is 1588 nucleotides (nt) long and encodes a 509 amino acid (aa) protein. The HIRRV glycoprotein gene sequence comprises 1612 nt, coding for a 508 aa protein. In sequence comparisons of 15 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2–33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

Phylogeography of infectious haematopoietic necrosis virus in North America

DEFF Research Database (Denmark)

Kurath, G.; Garver, K.A.; Troyer, R.M.

2003-01-01

Infectious hematopoietic necrosis virus (IHNV) is a rhabdoviral pathogen that infects wild and cultured salmonid fish throughout the Pacific Northwest of North America. IHNV causes severe epidemics in young fish and can cause disease or occur asymptomatically in adults. In a broad survey of 323...... IHNV field isolates, sequence analysis of a 303 nucleotide variable region within the glycoprotein gene revealed a maximum nucleotide diversity of 8(.)6%, indicating low genetic diversity overall for this virus. Phylogenetic analysis revealed three major virus genogroups, designated U, M and L, which...... varied in topography and geographical range. Intragenogroup genetic. diversity measures indicated that the M genogroup had three- to fourfold more diversity than the other genogroups and suggested relatively rapid evolution of the M genogroup and stasis within the U genogroup. We speculate that factors...

Computer-aided vaccine designing approach against fish pathogens Edwardsiella tarda and Flavobacterium columnare using bioinformatics softwares

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Mahendran R

2016-05-01

Full Text Available Radha Mahendran,1 Suganya Jeyabaskar,1 Gayathri Sitharaman,1 Rajamani Dinakaran Michael,2 Agnal Vincent Paul1 1Department of Bioinformatics, 2Centre for Fish Immunology, School of Life Sciences, Vels University, Pallavaram, Chennai, Tamil Nadu, India Abstract: Edwardsiella tarda and Flavobacterium columnare are two important intracellular pathogenic bacteria that cause the infectious diseases edwardsiellosis and columnaris in wild and cultured fish. Prediction of major histocompatibility complex (MHC binding is an important issue in T-cell epitope prediction. In a healthy immune system, the T-cells must recognize epitopes and induce the immune response. In this study, T-cell epitopes were predicted by using in silico immunoinformatics approach with the help of bioinformatics tools that are less expensive and are not time consuming. Such identification of binding interaction between peptides and MHC alleles aids in the discovery of new peptide vaccines. We have reported the potential peptides chosen from the outer membrane proteins (OMPs of E. tarda and F. columnare, which interact well with MHC class I alleles. OMPs from E. tarda and F. columnare were selected and analyzed based on their antigenic and immunogenic properties. The OMPs of the genes TolC and FCOL_04620, respectively, from E. tarda and F. columnare were taken for study. Finally, two epitopes from the OMP of E. tarda exhibited excellent protein–peptide interaction when docked with MHC class I alleles. Five epitopes from the OMP of F. columnare had good protein–peptide interaction when docked with MHC class I alleles. Further in vitro studies can aid in the development of potential peptide vaccines using the predicted peptides. Keywords: E. tarda, F. columnare, edwardsiellosis, columnaris, T-cell epitopes, MHC class I, peptide vaccine, outer membrane proteins 

Modelling Infectious Hematopoietic Necrosis Virus Dispersion from Marine Salmon Farms in the Discovery Islands, British Columbia, Canada.

Directory of Open Access Journals (Sweden)

Michael G G Foreman

Full Text Available Finite volume ocean circulation and particle tracking models are used to simulate water-borne transmission of infectious hematopoietic necrosis virus (IHNV among Atlantic salmon (Salmo salar farms in the Discovery Islands region of British Columbia, Canada. Historical simulations for April and July 2010 are carried out to demonstrate the seasonal impact of river discharge, wind, ultra-violet (UV radiation, and heat flux conditions on near-surface currents, viral dispersion and survival. Numerical particles released from infected farm fish in accordance with IHNV shedding rates estimated through laboratory experiments are dispersed by model oceanic flows. Viral particles are inactivated by ambient UV radiation levels and by the natural microbial community at rates derived through laboratory studies. Viral concentration maps showing temporal and spatial changes are produced and combined with lab-determined minimum infectious dosages to estimate the infective connectivity among farms. Results demonstrate that neighbouring naïve farms can become exposed to IHNV via water-borne transport from an IHNV diseased farm, with a higher risk in April than July, and that many events in the sequence of farm outbreaks in 2001-2002 are consistent with higher risks in our farm connectivity matrix. Applications to other diseases, transfers between farmed and wild fish, and the effect of vaccinations are also discussed.

Cloning and characterization of a new cold-adapted and thermo-tolerant ι-carrageenase from marine bacterium Flavobacterium sp. YS-80-122.

Science.gov (United States)

Li, Shangyong; Hao, Jianhua; Sun, Mi

2017-09-01

ι-Carrageenases play a role in marine ι-carrageenan degradation, and their enzymatic hydrolysates are thought to be excellent antioxidants. In this study, we identified a new ι-carrageenase, encoded by cgiF, in psychrophilic bacterium Flavobacterium sp. YS-80-122. The deduced ι-carrageenase, CgiF, belongs to glycoside hydrolase family 82 and shows less than 40% amino acid identity with characterized ι-carrageenases. The activity of recombinant CgiF peaked at 30°C (1,207.8U/mg). Notably, CgiF is a cold-adapted ι-carrageenase, which showed 36.5% and 57% of the maximum activity at 10°C and 15°C, respectively. In addition, it is a thermo-tolerant enzyme that recovered 58.2% of its initial activity after heat shock. Furthermore, although the activity of CgiF was enhanced by NaCl, the enzyme is active in absence of NaCl. This study also shows that CgiF is an endo-type ι-carrageenase that hydrolyzes β-1,4-linkages of ι-carrageenan, yielding neo-ι-carratetraose as the main product. Its cold-adaptation, thermo-tolerance, NaCl independence and high neo-ι-carratetraose yield make CgiF an excellent candidate for industrial applications in production of ι-carrageen oligosaccharides from seaweed polysaccharides. Copyright © 2017. Published by Elsevier B.V.

Spatial and temporal heterogeneity of infectious hematopoietic necrosis virus in Pacific Northwest salmonids

Science.gov (United States)

Breyta, Rachel; Black, Allison; Kaufman, John; Kurath, Gael

2016-01-01

The aquatic rhaboviral pathogen infectious hematopoietic necrosis virus (IHNV) causes acute disease in juvenile fish of a number of populations of Pacific salmonid species. Heavily managed in both marine and freshwater environments, these fish species are cultured during the juvenile stage in freshwater conservation hatcheries, where IHNV is one of the top three infectious diseases that cause serious morbidity and mortality. Therefore, a comprehensive study of viral genetic surveillance data representing 2590 field isolates collected between 1958 and 2014 was conducted to determine the spatial and temporal patterns of IHNV in the Pacific Northwest of the contiguous United States. Prevalence of infection varied over time, fluctuating over a rough 5–7 year cycle. The genetic analysis revealed numerous subgroups of IHNV, each of which exhibited spatial heterogeneity. Within all subgroups, dominant genetic types were apparent, though the temporal patterns of emergence of these types varied among subgroups. Finally, the affinity or fidelity of subgroups to specific host species also varied, where UC subgroup viruses exhibited a more generalist profile and all other subgroups exhibited a specialist profile. These complex patterns are likely synergistically driven by numerous ecological, pathobiological, and anthropogenic factors. Since only a few anthropogenic factors are candidates for managed intervention aimed at improving the health of threatened or endangered salmonid fish populations, determining the relative impact of these factors is a high priority for future studies.

Rainbow trout fed diets with varying content of marine and plant origin; how does that influence the outcome of experimental infections of the fry with Flavobacterium psychrophilum and Yersinia ruckeri?

DEFF Research Database (Denmark)

Madsen, Lone; Ingerslev, Hans-Christian; Boye, Mette

2014-01-01

disease caused by Yersinia ruckeri is also an economically important disease which causes problems in rainbow trout fry as well as larger fish. Rainbow trout were fed from first-feeding with five different diets; diet A with marine fish oil (conventional fry diet), diet B (an organic version of A), diet C......Feed for rainbow trout aquaculture has traditionally been based on marine resources such as fish meal and fish oil. Because of a shortage of marine resources as well as the growing production of farmed fish, the feed industry has been forced to partially exchange fish meal protein with proteins...... with rape seed oil (like B but with rape seed oil exchanging marine fish oil), diet C with pea protein (like B but added pea protein) and diet E with rape seed oil and pea protein. When the fry had reached sizes 1.5 g and 4 g, groups of fish from the five diet groups were infected with Flavobacterium...

Stress Tolerance-Related Genetic Traits of Fish Pathogen Flavobacterium psychrophilum in a Mature Biofilm

Directory of Open Access Journals (Sweden)

Héctor A. Levipan

2018-01-01

Full Text Available Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout fry syndrome, and hence this bacterium is placed among the most important salmonid pathogens in the freshwater aquaculture industry. Since bacteria in biofilms differ substantially from free-living counterparts, this study sought to find the main differences in gene expression between sessile and planktonic states of F. psychrophilum LM-02-Fp and NCMB1947T, with focus on stress-related changes in gene expression occurring during biofilm formation. To this end, biofilm and planktonic samples were analyzed by RNA sequencing to detect differentially expressed candidate genes (DECGs between the two growth states, and decreasing the effects of interstrain variation by considering only genes with log2-fold changes ≤ −2 and ≥ 2 at Padj-values ≤ 0.001 as DECGs. Overall, 349 genes accounting for ~15% of total number of genes expressed in transcriptomes of F. psychrophilum LM-02-Fp and NCMB1947T (n = 2327 were DECGs between biofilm and planktonic states. Approximately 83 and 81% of all up- and down-regulated candidate genes in mature biofilms, respectively, were assigned to at least one gene ontology term; these were primarily associated with the molecular function term “catalytic activity.” We detected a potential stress response in mature biofilms, characterized by a generalized down-regulation of DECGs with roles in the protein synthesis machinery (n = 63, primarily ribosomal proteins and energy conservation (seven ATP synthase subunit genes, as well as an up-regulation of DECGs involved in DNA repair (ruvC, recO, phrB1, smf, and dnaQ and oxidative stress response (cytochrome C peroxidase, probable peroxiredoxin, and a probable thioredoxin. These results support the idea of a strategic trade-off between growth-related processes and cell homeostasis to preserve biofilm structure and metabolic functioning. In addition, LDH-based cytotoxicity

The complete genome structure and phylogenetic relationship of infectious hematopoietic necrosis virus

Science.gov (United States)

Morzunov , Sergey P.; Winton, James R.; Nichol, Stuart T.

1995-01-01

Infectious hematopoietic necrosis virus (IHNV), a member of the family Rhabdoviridae, causes a severe disease with high mortality in salmonid fish. The nucleotide sequence (11, 131 bases) of the entire genome was determined for the pathogenic WRAC strain of IHNV from southern Idaho. This allowed detailed analysis of all 6 genes, the deduced amino acid sequences of their encoded proteins, and important control motifs including leader, trailer and gene junction regions. Sequence analysis revealed that the 6 virus genes are located along the genome in the 3′ to 5′ order: nucleocapsid (N), polymerase-associated phosphoprotein (P or M1), matrix protein (M or M2), surface glycoprotein (G), a unique non-virion protein (NV) and virus polymerase (L). The IHNV genome RNA was found to have highly complementary termini (15 of 16 nucleotides). The gene junction regions display the highly conserved sequence UCURUC(U)7RCCGUG(N)4CACR (in the vRNA sense), which includes the typical rhabdovirus transcription termination/polyadenylation signal and a novel putative transcription initiation signal. Phylogenetic analysis of M, G and L protein sequences allowed insights into the evolutionary and taxonomic relationship of rhabdoviruses of fish relative to those of insects or mammals, and a broader sense of the relationship of non-segmented negative-strand RNA viruses. Based on these data, a new genus, piscivirus, is proposed which will initially contain IHNV, viral hemorrhagic septicemia virus and Hirame rhabdovirus.

Protection of rainbow trout against infectious hematopoietic necrosis virus four days after specific or semi-specific DNA vaccination

DEFF Research Database (Denmark)

LaPatra, S.E.; Corbeil, S.; Jones, G.R.

2001-01-01

A DNA vaccine against a fish rhabdovirus, infectious hematopoietic necrosis virus (IHNV), was shown to provide significant protection as soon as 4 d after intramuscular vaccination in 2 g rainbow trout (Oncorhynchus mykiss) held at 15 degreesC. Nearly complete protection was also observed at late......-protection against IHNV challenge for a transient period of time, whereas a rabies virus DNA vaccine was not protective. This indication of distinct early and late protective mechanisms was not dependent on DNA vaccine doses from 0.1 to 2.5 mug....

Genetic diversity and epidemiology of infectious hematopoietic necrosis virus in Alaska

Science.gov (United States)

Emmenegger, E.G; Meyers, T.R.; Burton, T.O.; Kurath, G.

2000-01-01

Forty-two infectious hematopoietic necrosis virus (IHNV) isolates from Alaska were analyzed using the ribonuclease protection assay (RPA) and nucleotide sequencing. RPA analyses, utilizing 4 probes, N5, N3 (N gene), GF (G gene), and NV (NV gene), determined that the haplotypes of all 3 genes demonstrated a consistent spatial pattern. Virus isolates belonging to the most common haplotype groups were distributed throughout Alaska, whereas isolates in small haplotype groups were obtained from only 1 site (hatchery, lake, etc.). The temporal pattern of the GF haplotypes suggested a 'genetic acclimation' of the G gene, possibly due to positive selection on the glycoprotein. A pairwise comparison of the sequence data determined that the maximum nucleotide diversity of the isolates was 2.75% (10 mismatches) for the NV gene, and 1.99% (6 mismatches) for a 301 base pair region of the G gene, indicating that the genetic diversity of IHNV within Alaska is notably lower than in the more southern portions of the IHNV North American range. Phylogenetic analysis of representative Alaskan sequences and sequences of 12 previously characterized IHNV strains from Washington, Oregon, Idaho, California (USA) and British Columbia (Canada) distinguished the isolates into clusters that correlated with geographic origin and indicated that the Alaskan and British Columbia isolates may have a common viral ancestral lineage. Comparisons of multiple isolates from the same site provided epidemiological insights into viral transmission patterns and indicated that viral evolution, viral introduction, and genetic stasis were the mechanisms involved with IHN virus population dynamics in Alaska. The examples of genetic stasis and the overall low sequence heterogeneity of the Alaskan isolates suggested that they are evolutionarily constrained. This study establishes a baseline of genetic fingerprint patterns and sequence groups representing the genetic diversity of Alaskan IHNV isolates. This

Infectious Hematopoietic Necrosis Virus Transmission and Disease among Juvenile Chinook Salmon Exposed in Culture Compared to Environmentally Relevant Conditions

Directory of Open Access Journals (Sweden)

J. Scott Foott

2006-02-01

Full Text Available The dynamics of IHNV infection and disease were followed in a juvenile Chinook salmon population both during hatchery rearing and for two weeks post-release. Cumulative weekly mortality increased from 0.03%–3.5% as the prevalence of viral infection increased from 2%–22% over the same four-week period. The majority of the infected salmon was asymptomatic. Salmon demonstrating clinical signs of infection shed 1000 pfu mL-1 of virus into the water during a 1 min observation period and had a mean concentration of 106 pfu mL-1 in their mucus. The high virus concentration detected in mucus suggests that it could act as an avenue of transmission in high density situations where dominance behavior results in nipping. Infected smolts that had migrated 295 km down river were collected at least two weeks after their release. The majority of the virus positive smolts was asymptomatic. A series of transmission experiments was conducted using oral application of the virus to simulate nipping, brief low dose waterborne challenges, and cohabitation with different ratios of infected to naïve fish. These studies showed that asymptomatic infections will occur when a salmon is exposed for as little as 1 min to >102 pfu mL-1, yet progression to clinical disease is infrequent unless the challenge dose is >104 pfu mL-1. Asymptomatic infections were detected up to 39 d post-challenge. No virus was detected by tissue culture in natural Chinook juveniles cohabitated with experimentally IHNV-infected hatchery Chinook at ratios of 1:1, 1:10, and 1:20 for either 5 min or 24 h. Horizontal transmission of the Sacramento River strain of IHNV from infected juvenile hatchery fish to wild cohorts would appear to be a low ecological risk. The study results demonstrate key differences between IHNV infections as present in a hatchery and the natural environment. These differences should be considered during risk assessments of the impact of IHNV infections on wild salmon and

Detection and phylogenetic analysis of infectious pancreatic necrosis virus in Chile.

Science.gov (United States)

Tapia, D; Eissler, Y; Torres, P; Jorquera, E; Espinoza, J C; Kuznar, J

2015-10-27

Infectious pancreatic necrosis virus (IPNV) is the etiological agent of a highly contagious disease that is endemic to salmon farming in Chile and causes great economic losses to the industry. Here we compared different diagnostic methods to detect IPNV in field samples, including 3 real-time reverse transcription PCR (qRT-PCR) assays, cell culture isolation, and indirect fluorescent antibody test (IFAT). Additionally, we performed a phylogenetic analysis to investigate the genogroups prevailing in Chile, as well as their geographic distribution and virulence. The 3 qRT-PCR assays used primers that targeted regions of the VP2 and VP1 genes of the virus and were tested in 46 samples, presenting a fair agreement within their results. All samples were positive for at least 2 of the qRT-PCR assays, 29 were positive for cell culture, and 23 for IFAT, showing less sensitivity for these latter 2 methods. For the phylogenetic analysis, portions of 1180 and 523 bp of the VP2 region of segment A were amplified by RT-PCR, sequenced and compared with sequences from reference strains and from isolates reported by previous studies carried out in Chile. Most of the sequenced isolates belonged to genogroup 5 (European origin), and 5 were classified within genogroup 1 (American origin). Chilean isolates formed clusters within each of the genogroups found, evidencing a clear differentiation from the reference strains. To our knowledge, this is the most extensive study completed for IPNV in Chile, covering isolates from sea- and freshwater salmon farms and showing a high prevalence of this virus in the country.

Protective immunity and lack of histopathological damage two years after DNA vaccination against infectious hematopoietic necrosis virus in trout

Science.gov (United States)

Kurath, Gael; Garver, Kyle A.; Corbeil, Serge; Elliott, Diane G.; Anderson, Eric D.; LaPatra, Scott E.

2006-01-01

The DNA vaccine pIHNw-G encodes the glycoprotein of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV). Vaccine performance in rainbow trout was measured 3, 6, 13, 24, and 25 months after vaccination. At three months all fish vaccinated with 0.1 μg pIHNw-G had detectable neutralizing antibody (NAb) and they were completely protected from lethal IHNV challenge with a relative percent survival (RPS) of 100% compared to control fish. Viral challenges at 6, 13, 24, and 25 months post-vaccination showed protection with RPS values of 47–69%, while NAb seroprevalence declined to undetectable levels. Passive transfer experiments with sera from fish after two years post-vaccination were inconsistent but significant protection was observed in some cases. The long-term duration of protection observed here defined a third temporal phase in the immune response to IHNV DNA vaccination, characterized by reduced but significant levels of protection, and decline or absence of detectable NAb titers. Examination of multiple tissues showed an absence of detectable long-term histopathological damage due to DNA vaccination.

Immunity to rhabdoviruses in rainbow trout: the antibody response

DEFF Research Database (Denmark)

Lorenzen, Niels; Lapatra, S.E.

1999-01-01

to their occasional detrimental effect on rainbow trout farming. Research efforts have been focused on understanding the mechanisms involved in protective immunity. Several specific and nonspecific cellular and humoral parameters are believed to be involved, but only the antibody response has been characterised......, have demonstrated that rainbow trout can produce specific and highly functional antibodies that are able to neutralise virus pathogenicity in vitro as well as in vivo. The apparently more restricted antibody response to IHNV and VHSV antigens in fish compared to mammals could possibly be explained...... aspects of antibody response and antibody reactivity with IHNV and VHSV antigens....

Effect of Pseudomonas sp. MT5 baths on Flavobacterium columnare infection of rainbow trout and on microbial diversity on fish skin and gills.

Science.gov (United States)

Suomalainen, L R; Tiirola, M A; Valtonen, E T

2005-01-25

Use of Pseudomonas sp. strain MT5 to prevent and treat Flavobacterium columnare infection was studied in 2 experiments with fingerling rainbow trout Oncorhynchus mykiss. In the first experiment, length heterogeneity analysis of PCR-amplified DNA fragments (LH-PCR) was used to assess the effect of antagonistic baths on the microbial diversity of healthy and experimentally infected fish. In the 148 samples studied, no difference was found between bathed and unbathed fish, and 3 fragment lengths were detected most frequently: 500 (in 75.7% of the samples), 523 (62.2%) and 517 bp (40.5%). The species contributing to these fragment sizes were Pseudomonas sp., Rhodococcus sp. and F. columnare, respectively. A specific PCR for detection of Pseudomonas sp. MT5 was designed, but none of the tissue samples were found to be positive, most likely indicating poor adhesion of the strain during bathing. LH-PCR was found to be a more powerful tool for detecting F. columnare in fish tissue than traditional culture methods (chi2 = 3.9, df = 1, p < 0.05). Antagonistic baths had no effect on the outbreak of infection or on fish mortality. F. columnare was also detected in healthy fish prior to and after experimental infection, indicating that these fish were carriers of the disease. In the second experiment, intensive Pseudomonas sp. MT5 antagonistic baths were given daily to rainbow trout suffering from a natural columnaris infection. Again, the antagonistic bacteria had no effect on fish mortality, which reached 95 % in both control and antagonist-treated groups in 7 d.

Stabilization of microorganisms for in situ degradation of toxic chemicals

International Nuclear Information System (INIS)

Crawford, R.L.; Ralston, D.R.

1993-01-01

Methods for large-scale microencapsulation of bacteria and nutrients into microbeads with small enough diameters to travel through aquifers have been developed at the University of Idaho. Both free and immobilized cells of Flavobacterium ATCC 39723, a gram-negative aerobe that degrades various chlorinated phenols, into aquifer microcosms, through which pentachlorophenol (PCP)-contaminated groundwater flowed at in situ flow rates. Aquifer samples were collected with an auger from three wells at the University of Idaho Ground Water Research Site, and packed into 24 columns. Some sterile columns were also prepared, by irradiation at the Washington State University Radiation Center. In some of the columns the free Flavobacterium cells were mixed with the aquifer material before packing the columns. In others, agarose-microimmobilized Flavobacterium were mixed into the aquifer material. The effluent from each column was collected daily for 170 days and analyzed by UV spectroscopy or HPLC for remaining PCP. There were no statistically significant differences between the degradation rates of free or encapsulated Flavobacterium in sterile or native aquifer material as tested in these experiments. This work has shown at the lab scale that encapsulated PCP-degrading Flavobacterium were able to survive under conditions of starvation, predation, and lack of water

Comparison of vaccine efficacy for different antigen delivery systems for infectious pancreatic necrosis virus vaccines in Atlantic salmon (Salmo salar L.) in a cohabitation challenge model.

Science.gov (United States)

Munang'andu, Hetron M; Fredriksen, Børge N; Mutoloki, Stephen; Brudeseth, Bjørn; Kuo, Tsun-Yung; Marjara, Inderjit S; Dalmo, Roy A; Evensen, Øystein

2012-06-08

Two strains of IPNV made by reverse genetics on the Norwegian Sp strain NVI-015 (GenBank AY379740) backbone encoding the virulent (T(217)A(221)) and avirulent (P(217)T(221)) motifs were used to prepare inactivated whole virus (IWV), nanoparticle vaccines with whole virus, Escherichia coli subunit encoding truncated VP2-TA and VP2-PT, VP2-TA and VP2-PT fusion antigens with putative translocating domains of Pseudomonas aeruginosa exotoxin, and plasmid DNA encoding segment A of the TA strain. Post challenge survival percentages (PCSP) showed that IWV vaccines conferred highest protection (PCSP=42-53) while nanoparticle, sub-unit recombinant and DNA vaccines fell short of the IWV vaccines in Atlantic salmon (Salmo salar L.) postsmolts challenged with the highly virulent Sp strain NVI-015 (TA strain) of IPNV after 560 degree days post vaccination. Antibody levels induced by these vaccines did not show antigenic differences between the virulent and avirulent motifs for vaccines made with the same antigen dose and delivery system after 8 weeks post vaccination. Our findings show that fish vaccinated with less potent vaccines comprising of nanoparticle, DNA and recombinant vaccines got infected much earlier and yielded to higher infection rates than fish vaccinated with IWV vaccines that were highly potent. Ability of the virulent (T(217)A(221)) and avirulent (P(217)T(221)) motifs to limit establishment of infection showed equal protection for vaccines made of the same antigen dose and delivery systems. Prevention of tissue damage linked to viral infection was eminent in the more potent vaccines than the less protective ones. Hence, there still remains the challenge of developing highly efficacious vaccines with the ability to eliminate the post challenge carrier state in IPNV vaccinology. Copyright © 2012 Elsevier Ltd. All rights reserved.

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout, Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum.

Science.gov (United States)

Ryerse, I A; Hooft, J M; Bureau, D P; Hayes, M A; Lumsden, J S

2015-09-01

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10(6) colony-forming units [CFU] mL(-1) ). Mortality of rainbow trout fed either 6.4 mg kg(-1) DON or trout pair-fed the control diet was significantly reduced (P trout fed the control diet to apparent satiation (trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg(-1) DON) or a diet containing purified DON (3.8 mg kg(-1) ); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P trout fed diets containing purified DON (3.8 mg kg(-1) ) was significantly higher (P < 0.05) at 35 day post-exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L(-1) DON, but no effect was observed below this concentration (0-30 mg L(-1) ). © 2014 John Wiley & Sons Ltd.

Development of a biotinylated DNA probe for detection of infectious hematopoietic necrosis virus

Science.gov (United States)

Deering, R.E.; Arakawa, C.K.; Oshima, K.H.; O'Hara, P.J.; Landolt, M.L.; Winton, J.R.

1991-01-01

A nonrad~oact~ve DNA probe assay was developed to detect and ~dent~fy infect~ous hernatopoiet~c necrosls virus (IHNV) uslng a dot blot format The probe a synthet~c DNA oligonucleot~de labeled enzymatlcally w~th biotln hybnd~zed spec~f~cally w~th nucleocaps~d mRNA extracted from Infected cells early In the vlrus repl~cation cycle A rap~d guan~dln~um th~ocyanate based RNA extraction method uslng RNAzol B and rn~crocentrifuge tubes eff~c~ently pioduced h~gh qual~ty RNA from 3 commonly used f~sh cell llnes, CHSE-214, CHH-1, and EPC The probe reacted with 6 d~verse ~solates of IHNV, but d~d not react \

In Vivo fitness associated with high virulence in a vertebrate virus is a complex trait regulated by host entry, replication, and shedding

Science.gov (United States)

Wargo, Andrew R.; Kurath, Gael

2011-01-01

The relationship between pathogen fitness and virulence is typically examined by quantifying only one or two pathogen fitness traits. More specifically, it is regularly assumed that within-host replication, as a precursor to transmission, is the driving force behind virulence. In reality, many traits contribute to pathogen fitness, and each trait could drive the evolution of virulence in different ways. Here, we independently quantified four viral infection cycle traits, namely, host entry, within-host replication, within-host coinfection fitness, and shedding, in vivo, in the vertebrate virus Infectious hematopoietic necrosis virus (IHNV). We examined how each of these stages of the viral infection cycle contributes to the fitness of IHNV genotypes that differ in virulence in rainbow trout. This enabled us to determine how infection cycle fitness traits are independently associated with virulence. We found that viral fitness was independently regulated by each of the traits examined, with the largest impact on fitness being provided by within-host replication. Furthermore, the more virulent of the two genotypes of IHNV we used had advantages in all of the traits quantified. Our results are thus congruent with the assumption that virulence and within-host replication are correlated but suggest that infection cycle fitness is complex and that replication is not the only trait associated with virulence.

Atlantic salmon, Salmo salar L. are broadly susceptible to isolates representing the North American genogroups of infectious hematopoietic necrosis virus

Science.gov (United States)

Kurath, Gael; Winton, James R.; Dale, Ole B.; Purcell, Maureen K.; Falk, Knut; Busch, Robert D.

2016-01-01

Beginning in 1992, three epidemic waves of infectious hematopoietic necrosis, often with high mortality, occurred in farmed Atlantic salmon Salmo salar L. on the west coast of North America. We compared the virulence of eleven strains of infectious hematopoietic necrosis virus (IHNV), representing the U, M and L genogroups, in experimental challenges of juvenile Atlantic salmon in freshwater. All strains caused mortality and there was wide variation within genogroups: cumulative mortality for five U-group strains ranged from 20 to 100%, four M-group strains ranged 30-63% and two L-group strains varied from 41 to 81%. Thus, unlike Pacific salmonids, there was no apparent correlation of virulence in a particular host species with virus genogroup. The mortality patterns indicated two different phenotypes in terms of kinetics of disease progression and final per cent mortality, with nine strains having moderate virulence and two strains (from the U and L genogroups) having high virulence. These phenotypes were investigated by histopathology and immunohistochemistry to describe the variation in the course of IHNV disease in Atlantic salmon. The results from this study demonstrate that IHNV may become a major threat to farmed Atlantic salmon in other regions of the world where the virus has been, or may be, introduced.

Oral DNA vaccination of rainbow trout, Oncorhynchus mykiss (Walbaum), against infectious haematopoietic necrosis virus using PLGA [Poly(D,L-Lactic-Co-Glycolic Acid)] nanoparticles.

Science.gov (United States)

Adomako, M; St-Hilaire, S; Zheng, Y; Eley, J; Marcum, R D; Sealey, W; Donahower, B C; Lapatra, S; Sheridan, P P

2012-03-01

A DNA vaccine against infectious haematopoietic necrosis virus (IHNV) is effective at protecting rainbow trout, Oncorhynchus mykiss, against disease, but intramuscular injection is required and makes the vaccine impractical for use in the freshwater rainbow trout farming industry. Poly (D,L-lactic-co-glycolic acid) (PLGA) is a U.S. Food and Drug Administration (FDA) approved polymer that can be used to deliver DNA vaccines. We evaluated the in vivo absorption of PLGA nanoparticles containing coumarin-6 when added to a fish food pellet. We demonstrated that rainbow trout will eat PLGA nanoparticle coated feed and that these nanoparticles can be detected in the epithelial cells of the lower intestine within 96 h after feeding. We also detected low levels of gene expression and anti-IHNV neutralizing antibodies when fish were fed or intubated with PLGA nanoparticles containing IHNV G gene plasmid. A virus challenge evaluation suggested a slight increase in survival at 6 weeks post-vaccination in fish that received a high dose of the oral vaccine, but there was no difference when additional fish were challenged at 10 weeks post-vaccination. The results of this study suggest that it is possible to induce an immune response using an orally delivered DNA vaccine, but the current system needs improvement. © 2012 Blackwell Publishing Ltd.

Multiplication of infectious hematopoietic necrosis virus in rainbow trout following immersion infection: whole-body assay and immunohistochemistry

Science.gov (United States)

Yamamoto, T.; Batts, W.N.; Arakawa, C.K.; Winton, J.R.

1990-01-01

The sites of replication of infectious hematopoietic necrosis virus (IHNV) in infected tissues were detected in fingerling rainbow trout Oncorhynchus mykiss by in situ histologic techniques following immersion infection. Virus antigens in tissues were detected by a neutralizing mouse monoclonal antibody and a one-step anti-mouse biotin-streptavidin conjugated to horseradish peroxidase. The efficiency of infection and virulence of the virus determined by mortality rates showed high virulence of the selected IHNV isolates, and viral replication in individual fish showed that virus content of the fish increased rapidly from the second day to the seventh day postinfection. The earliest viral lesions following infection were detected in the epidermis of the pectoral fins, opercula, and ventral surface of the body. Virus lesions became evident in kidneys on the third day. By the fifth day, when there was a significant increase in virus titer, foci of viral replication were detected in gill tissue and in the anterior internal tissues below the epidermis. Subsequently, extensive virus replication and tissue destruction were observed in the spleen, dorsal adipose tissues, ventricle, and pseudobranch. Replication in the liver, the muscularis layers of the digestive tract, and the general body musculature followed later. These infection experiments indicated that the epidermis and gills of fish constitute important sites of early IHNV replication.

Flavobacterium capsulatum

African Journals Online (AJOL)

long on the human skin surface. ... It produces septicaemia with serious lung and brain damage in the newborn, ... aches, tiredness and lack ofappetite, and lost IQ kg in weight. In ... globulin, complement and auto-antibody levels and full blood.

Efficacy of a polyvalent immersion vaccine against Flavobacterium psychrophilum and evaluation of immune response to vaccination in rainbow trout fry (Onchorynchus mykiss L.).

Science.gov (United States)

Hoare, R; Ngo, T P H; Bartie, K L; Adams, A

2017-08-18

Rainbow trout fry syndrome (RTFS) is a disease caused by the Gram-negative bacterium Flavobacterium psychrophilum, responsible for significant economic losses in salmonid aquaculture worldwide. The diversity of F. psychrophilum isolates and the inherent difficulties in vaccinating juvenile fish has hampered the development of a vaccine for RTFS. Disease episodes tend to occur between 10-14 °C with necrotic lesions often seen on the skin surrounding the dorsal fin and tail. At present no commercial vaccines are available for RTFS in the UK, leaving antibiotics as the only course of action to control disease outbreaks. The current work was performed as a pilot study to assess the efficacy of a polyvalent, whole cell vaccine containing formalin-inactivated F. psychrophilum, to induce protective immunity in rainbow trout fry. Duplicate groups of 30 trout (5 g) were immersed in 1 L of the vaccine for 30 s. Samples were taken 4 h, day 2 and 7 post-vaccination (pv) of skin mucus, tissues for histology and gene expression analysis; serum and histology samples were taken 6 weeks pv. A booster vaccination was given at 315 degree days (dd) also by immersion. Challenge was by immersion with a heterologous isolate of F. psychrophilum 630 dd post primary vaccination. The vaccine provided significant protection to the trout fry with a RPS of 84% (p < 0.0001). Detection of increased numbers of IgT positive cells in systemic organs, up-regulation of IgT expression in hind-gut and an increase in total IgT in serum was observed in vaccinated fish; however a functional role of IgT in the observed protection remains to be demonstrated.

Occurrence of different types of infectious hematopoietic necrosis virus in fish

International Nuclear Information System (INIS)

Hsu, Y.; Engelking, H.M.; Leong, J.C.

1986-01-01

The virion protein patterns of 71 isolates of infectious hematopoietic necrosis virus (IHNV) from the Pacific Northwest were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [ 35 S]-methionine-labeled virus. This analysis led to the classification of these virus isolates into four or more types. Type 1 virus was characterized by a nucleocapsid protein with an approximate molecular weight of 40,500. Type 2 and type 3 viruses have nucleocapsid proteins with molecular weights of 42,800 and 43,250, respectively. Type 2 virus was responsible for the recent epizootics of IHNV among fish in the lower Columbia River. The California IHNV isolates were type 3 with the exception of some of those isolated from fish at the Coleman Hatchery on the Sacramento River. These Coleman Hatchery isolates belonged to a type 4 virus group characterized by a larger glycoprotein of approximately 70,000 molecular weight. All other viruses examined had glycoproteins of 67,000 molecular weight. The type 5 virus isolates were grouped together because they were not sufficiently distinct to warrant classification into a separate type. These findings have been useful in determining that (i) a particular virus type is characteristic for a geographic area and will infect many different salmonid species in that area and (ii) the same type isolated from parental fish is responsible for the subsequent outbreak of the diseases in progeny

Infectious pancreatic necrosis virus in fish by-products is inactivated with inorganic acid (pH 1) and base (pH 12).

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Myrmel, M; Modahl, I; Nygaard, H; Lie, K M

2014-04-01

The aquaculture industry needs a simple, inexpensive and safe method for the treatment of fish waste without heat. Microbial inactivation by inorganic acid (HCl) or base (KOH) was determined using infectious pancreatic necrosis virus (IPNV) as a model organism for fish pathogens. Salmonella and spores of Clostridium perfringens were general hygiene indicators in supplementary examinations. IPNV, which is considered to be among the most chemical- and heat-resistant fish pathogens, was reduced by more than 3 log in 4 h at pH 1.0 and pH 12.0. Salmonella was rapidly inactivated by the same treatment, whereas spores of C. perfringens were hardly affected. The results indicate that low and high pH treatment could be particularly suitable for fish waste destined for biogas production. pH treatment at aquaculture production sites could reduce the spread of fish pathogens during storage and transportation without disturbing the anaerobic digestion process. The treatment could also be an alternative to the current energy-intensive steam pressure sterilization of fish waste to be used by the bioenergy, fertilizer and soil improver industries. © 2013 John Wiley & Sons Ltd.

Hexamitiasis leads to lower metabolic rates in rainbow trout Oncorhynchus mykiss (Walbaum) juveniles.

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Ogut, H; Parlak, R

2014-12-01

This study assessed the effects of Hexamita salmonis (Moore) on metabolism of rainbow trout Oncorhynchus mykiss (Walbaum) and its effect on the host's susceptibility to infectious pancreatic necrosis virus (IPNV) after antiparasitic treatment. Rainbow trout naturally infected with H. salmonis were treated with 10 mg metronidazole kg fish(-1) per day, and their physiological recovery was assessed through measuring resting metabolism on the 7th, 14th, 21st and 28th day after treatment. In addition, we exposed the naïve fish to H. salmonis and measured the resting metabolism (oxygen consumption as mg O2 kg(-1) per hour) on the 10th, 20th and 30th day after the exposure to assess the variation in metabolic rates after infection. Significantly lower rates of metabolic activity (P trout to IPNV remained unchanged in the presence of H. salmonis. Weight loss was significantly higher (P < 0.05) in infected than that in the parasite-free fish. Fish should be examined regularly for H. salmonis and treated immediately whether found to prevent economic losses and excessive size variation. © 2013 John Wiley & Sons Ltd.

An epidemiological model of virus transmission in salmonid fishes of the Columbia River Basin

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Ferguson, Paige F. B.; Breyta, Rachel; Brito, Ilana L.; Kurath, Gael; LaDeau, Shannon L.

2018-01-01

We have developed a dynamic epidemiological model informed by records of viral presence and genotypes to evaluate potential transmission routes maintaining a viral pathogen in economically and culturally important anadromous fish populations. In the Columbia River Basin, infectious hematopoietic necrosis virus (IHNV) causes severe disease, predominantly in juvenile steelhead trout (Oncorhynchus mykiss) and less frequently in Chinook salmon (O. tshawytscha). Mortality events following IHNV infection can be devastating for individual hatchery programs. Despite reports of high local mortality and extensive surveillance efforts, there are questions about how viral transmission is maintained. Modeling this system offers important insights into disease transmission in natural aquatic systems, as well as about the data requirements for generating accurate estimates about transmission routes and infection probabilities. We simulated six scenarios in which testing rates and the relative importance of different transmission routes varied. The simulations demonstrated that the model accurately identified routes of transmission and inferred infection probabilities accurately when there was testing of all cohort-sites. When testing records were incomplete, the model accurately inferred which transmission routes exposed particular cohort-sites but generated biased infection probabilities given exposure. After validating the model and generating guidelines for result interpretation, we applied the model to data from 14 annual cohorts (2000–2013) at 24 focal sites in a sub-region of the Columbia River Basin, the lower Columbia River (LCR), to quantify the relative importance of potential transmission routes in this focal sub-region. We demonstrate that exposure to IHNV via the return migration of adult fish is an important route for maintaining IHNV in the LCR sub-region, and the probability of infection following this exposure was relatively high at 0.16. Although only 1% of

Hymenobacter roseosalivarius gen. nov., sp. nov. from continental Antartica soils and sandstone: bacteria of the Cytophaga/Flavobacterium/Bacteroides line of phylogenetic descent.

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Hirsch, P; Ludwig, W; Hethke, C; Sittig, M; Hoffmann, B; Gallikowski, C A

1998-08-01

Aseptically collected sandstone and soil samples from the antarctic Dry Valleys were inoculated into oligotrophic media and incubated under low light intensities. A total of 41 Gram-negative isolates were obtained with reddish colonies spreading on agar. A sandstone isolate and four soil strains were characterized further. They were nearly identical in morphological, physiological, biochemical and chemotaxonomic properties. They produced large amounts of extracellular polymer and utilized for growth: glucose, saccharose, mannitol, sorbitol, L-aspartate, malate and acetate, but not D-ribose, adonitol, DL-alanine, glutamate, glycolate, lactate or succinate. All strains hydrolyzed gelatin, starch, casein, xylan, Tweens 80 or 60 and dead or living yeast cells, but not cellulose or pectin. Nitrate was not reduced, ethanol was not oxidized and acid was not produced from maltose, mannitol or dulcitol. Ammonia was not produced from peptone. They were strictly aerobic. Major fatty acids were n 16:1 d 9, n 16:1 d 11, n 17:1 d 11, and i 15:0. The strains contained the quinone MK-7 and phosphatidylethanolamine as the main phospholipid. The base ratio ranged from 55 to 61 mol% G+C. A 16S rRNA sequence analysis of strains AA-688 and AA-718 showed these to be identical and to represent a special phylogenetic group within the Cytophaga/Flavobacterium/Bacteroides major line of descent. Three soil strains labeled "Taxeobacter" Txc1, Txg1, and Txo1 (Reichenbach, 1992) belonged to the same group but had lower sequence similarities (<95%). Some of their characteristics were different from those of the antarctic strains: the utilization of C-compounds, hydrolysis of polymers, temperature tolerances, major fatty acids and base ratios. Txc1 and Txg1 may later have to be considered as members of this group, possibly on the species level, while Txo1 could represent a different related genus. It is concluded that the five antarctic strains represent a new genus and species for which the name

Epidemiology and Control of Infectious Diseases of Salmonids in the Columbia River Basin, 1984 Annual Report.

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Fryer, John L.

1985-11-01

monitoring bacterial kidney disease in adults. Antigenic differences among strains of R. salmoninarum and common antigens present on both R. salmoninarum and other Gram positive bacteria have been demonstrated for the first time using monoclonal antibodies. All of the monoclonal antibodies belong to the murine IgGl, IgG3 or TgG2a class and subclass. Field studies at Round Butte Hatchery with the molecular filtration apparatus detected IHNV in effluent water from the adult holding pond and in water from a tank containing steelhead trout fry infected with IHN disease. The concentrations of IHNV detected in these samples suggested that in the order of 10{sup 10} virions are being released each day into the Deschutes River at the peak of steelhead trout spawning at Round Butte Hatchery. Isolation of IHNV from dead eggs suggested that virus replication during incubation may be a possible cause of egg mortality. Two possible reasons for inconsistencies in the data from the IHNV transmission studies at Round Butte Hatchery are: (1) UV treatment does not completely sterilize the water and (2) vertical transmission occurs but under, as yet, undescribed conditions. Constant IHNV production over a prolonged period has been recorded in unfiltered ovarian fluid samples. Filtration eliminates this virus production. These observations suggest that cellular components in ovarian fluid are responsible for producing the delayed appearance of IHNV after storage at 4 C for 8 to 16 days.

Epidemiology and Control of Infectious Diseases of Salmonids in the Columbia River Basin, 1983 Annual Report.

Energy Technology Data Exchange (ETDEWEB)

Fryer, John L.

1984-11-01

salmon seined from the Columbia River just before entering the estuary. Interpretation of these numbers suggests an even greater economic impact on Columbia River salmonid stocks than that proposed for C. shasta. Fertilized eggs from bacterial kidney disease infected parents examined after one month of incubation revealed the presence of bacteria with identical morphology to R. salmoninarum on or in the egg wall further reinforcing the proposed vertical transmission of this disease organism. Infectious hematopoietic necrosis virus was recovered at the 67% level from seeded water samples supplemented with 1% fetal calf serum. Virus injected into unfertilized eggs survived for over two weeks; in eyed eggs the virus also replicated. Epizootics caused by IHNV occurred in two of the 8 separate groups of steelhead trout fingerlings held in LJV treated water at Round Butte Hatchery. Comparing these results to those in the vertical transmission experiment where none of the groups developed IHNV suggests that vertical transmission of IHNV, if it occurs, is a very infrequent or random event. On three occasions IHNV was detected in ovarian fluid samples after storage for 6--9 days at 4 C. No virus had been detected in these samples at spawning. This suggests the presence of an interfering substance, perhaps anti-IHNV antibody in ovarian fluid. This observation raises the possibility that IHNV is much more widespread throughout Columbia River Basin salmonid stocks than previously believed.

Relationship between apoptosis and the BH2 domain sequence of the VP5 peptide of infectious pancreatic necrosis virus

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Cesar Ortega S.

2014-03-01

Full Text Available Objective. To determine whether the level of apoptosis induced by infectious pancreatic necrosis virus (IPNV is related to the amino acid sequence of the BH2 domain of the VP5 protein and the level of infectivity. Materials and methods. Three IPNV strains were used, the VP2 protein gene was amplified for genotyping and the VP5 sequence was also obtained. The infectivity of the strains was calculated using the viral titer obtained at 12, 24, 36 and 45 hpi in CHSE-214 cells. The percentage of apoptosis in infected cells was visualized by TUNEL assay and immunohistochemistry (caspase 3 detection. Results. The V70/06 and V33/98 strains corresponded to genotype Sp, while V112/06 to VR-299; the amino acid analysis of the V70/06 strain allows its classification as middle virulent strain and V33/98 and V112/06 strains as low virulent ones; infection with the V112/06 strain produced a lower viral titer (p0.05. Conclusions. The results showed that the differences in the BH2 sequence of the VP5 protein, infectivity and the VP2 sequence are not associated with the modulation of apoptosis.

A historical review of the key bacterial and viral pathogens of Scottish wild fish.

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Wallace, I S; McKay, P; Murray, A G

2017-12-01

Thousands of Scottish wild fish were screened for pathogens by Marine Scotland Science. A systematic review of published and unpublished data on six key pathogens (Renibacterium salmoninarum, Aeromonas salmonicida, IPNV, ISAV, SAV and VHSV) found in Scottish wild and farmed fish was undertaken. Despite many reported cases in farmed fish, there was a limited number of positive samples from Scottish wild fish, however, there was evidence for interactions between wild and farmed fish. A slightly elevated IPNV prevalence was reported in wild marine fish caught close to Atlantic salmon, Salmo salar L., farms that had undergone clinical IPN. Salmonid alphavirus was isolated from wild marine fish caught near Atlantic salmon farms with a SAV infection history. Isolations of VHSV were made from cleaner wrasse (Labridae) used on Scottish Atlantic salmon farms and VHSV was detected in local wild marine fish. However, these pathogens have been detected in wild marine fish caught remotely from aquaculture sites. These data suggest that despite the large number of samples taken, there is limited evidence for clinical disease in wild fish due to these pathogens (although BKD and furunculosis historically occurred) and they are likely to have had a minimal impact on Scottish wild fish. © 2017 Crown Copyright. Journal of Fish Diseases © 2017 John Wiley & Sons Ltd.

Antimicrobial Susceptibility of Flavobacterium psychrophilum from Chilean Salmon Farms and Their Epidemiological Cut-Off Values Using Agar Dilution and Disk Diffusion Methods.

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Miranda, Claudio D; Smith, Peter; Rojas, Rodrigo; Contreras-Lynch, Sergio; Vega, J M Alonso

2016-01-01

Flavobacterium psychrophilum is the most important bacterial pathogen for freshwater farmed salmonids in Chile. The aims of this study were to determine the susceptibility to antimicrobials used in fish farming of Chilean isolates and to calculate their epidemiological cut-off (CO WT ) values. A number of 125 Chilean isolates of F. psychrophilum were isolated from reared salmonids presenting clinical symptoms indicative of flavobacteriosis and their identities were confirmed by 16S rRNA polymerase chain reaction. Susceptibility to antibacterials was tested on diluted Mueller-Hinton by using an agar dilution MIC method and a disk diffusion method. The CO WT values calculated by Normalized Resistance Interpretation (NRI) analysis allow isolates to be categorized either as wild-type fully susceptible (WT) or as manifesting reduced susceptibility (NWT). When MIC data was used, NRI analysis calculated a CO WT of ≤0.125, ≤2, and ≤0.5 μg mL -1 for amoxicillin, florfenicol, and oxytetracycline, respectively. For the quinolones, the CO WT were ≤1, ≤0.5, and ≤0.125 μg mL -1 for oxolinic acid, flumequine, and enrofloxacin, respectively. The disk diffusion data sets obtained in this work were extremely diverse and were spread over a wide range. For the quinolones there was a close agreement between the frequencies of NWT isolates calculated using MIC and disk data. For oxolinic acid, flumequine, and enrofloxacin the frequencies were 45, 39, and 38% using MIC data, and 42, 41, and 44%, when disk data were used. There was less agreement with the other antimicrobials, because NWT frequencies obtained using MIC and disk data, respectively, were 24 and 10% for amoxicillin, 8 and 2% for florfenicol, and 70 and 64% for oxytetracycline. Considering that the MIC data was more precise than the disk diffusion data, MIC determination would be the preferred method for susceptibility testing for this species and the NWT frequencies derived from the MIC data sets should be

Viral fitness does not correlate with three genotype displacement events involving infectious hematopoietic necrosis virus

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Kell, Alison M.; Wargo, Andrew R.; Kurath, Gael

2014-01-01

Viral genotype displacement events are characterized by the replacement of a previously dominant virus genotype by a novel genotype of the same virus species in a given geographic region. We examine here the fitness of three pairs of infectious hematopoietic necrosis virus (IHNV) genotypes involved in three major genotype displacement events in Washington state over the last 30 years to determine whether increased virus fitness correlates with displacement. Fitness was assessed using in vivo assays to measure viral replication in single infection, simultaneous co-infection, and sequential superinfection in the natural host, steelhead trout. In addition, virion stability of each genotype was measured in freshwater and seawater environments at various temperatures. By these methods, we found no correlation between increased viral fitness and displacement in the field. These results suggest that other pressures likely exist in the field with important consequences for IHNV evolution.

Concentration of infectious aquatic rhabdoviruses from freshwater and seawater using ultrafiltration.

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Grant, Amelia A M; Jakob, Eva; Richard, Jon; Garver, Kyle A

2011-12-01

Infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus, and spring viremia of carp virus were concentrated and detected from freshwater and seawater samples by using hollow-fiber ultrafiltration. Within 60 min, virus in a 50-L freshwater or saltwater sample was concentrated more than 70-fold, and virus retention efficiencies were consistently greater than 88%. Retention efficiency was highly dependent upon concentrations of column blocking and sample stabilization solutions. A large column with a surface area of 1.15 m2 and a filtration capacity of 5-200 L exhibited optimal viral retention when blocked with 2% fetal bovine serum (FBS) and when the samples were supplemented with 0.1% FBS. Conversely, a small column with 100-fold less surface area and a filtering capacity of 0.5-2.0 L was optimized when blocked with 1% FBS and when the samples were supplemented with 0.1% FBS. The optimized ultrafiltration procedure was further validated with water from a tank that contained IHNV-exposed juvenile sockeye salmon Oncorhynchus nerka, resulting in an average virus retention efficiency of 91.6 +/- 4.1% (mean +/- SE). Virus quantification of concentrated samples demonstrated that IHNV shedding in sockeye salmon preceded mortality; shedding of the virus was observed to increase significantly as early as 7 d postchallenge and peaked at day 14, when virus levels reached 4.87 x 10(3) plaque-forming units/mL. We conclude that ultrafiltration is a reliable and effective method for concentrating viable aquatic rhabdoviruses from large volumes of water and has application for the analysis of environmental water samples.

Geography and host species shape the evolutionary dynamics of U genogroup infectious hematopoietic necrosis virus

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Black, Allison; Breyta, Rachel; Bedford, Trevor; Kurath, Gael

2016-01-01

Infectious hematopoietic necrosis virus (IHNV) is a negative-sense RNA virus that infects wild and cultured salmonids throughout the Pacific Coastal United States and Canada, from California to Alaska. Although infection of adult fish is usually asymptomatic, juvenile infections can result in high mortality events that impact salmon hatchery programs and commercial aquaculture. We used epidemiological case data and genetic sequence data from a 303 nt portion of the viral glycoprotein gene to study the evolutionary dynamics of U genogroup IHNV in the Pacific Northwestern United States from 1971 to 2013. We identified 114 unique genotypes among 1,219 U genogroup IHNV isolates representing 619 virus detection events. We found evidence for two previously unidentified, broad subgroups within the U genogroup, which we designated ‘UC’ and ‘UP’. Epidemiologic records indicated that UP viruses were detected more frequently in sockeye salmon (Oncorhynchus nerka) and in coastal waters of Washington and Oregon, whereas UC viruses were detected primarily in Chinook salmon (Oncorhynchus tshawytscha) and steelhead trout (Oncorhynchus mykiss) in the Columbia River Basin, which is a large, complex watershed extending throughout much of interior Washington, Oregon, and Idaho. These findings were supported by phylogenetic analysis and by FST. Ancestral state reconstruction indicated that early UC viruses in the Columbia River Basin initially infected sockeye salmon but then emerged via host shifts into Chinook salmon and steelhead trout sometime during the 1980s. We postulate that the development of these subgroups within U genogroup was driven by selection pressure for viral adaptation to Chinook salmon and steelhead trout within the Columbia River Basin.

Detection and quantification of Flavobacterium psychrophilum-specific bacteriophages in vivo in rainbow trout upon oral administration: implications for disease control in aquaculture.

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Christiansen, Rói Hammershaimb; Dalsgaard, Inger; Middelboe, Mathias; Lauritsen, Anne H; Madsen, Lone

2014-12-01

The use of bacteriophages in the treatment and prevention of infections by the fish pathogen Flavobacterium psychrophilum has attracted increased attention in recent years. It has been shown recently that phage delivery via the parenteral route resulted in immediate distribution of phages to the circulatory system and the different organs. However, little is known about phage dispersal and survival in vivo in rainbow trout after delivery via the oral route. Here we examined the dispersal and survival of F. psychrophilum phage FpV-9 in vivo in juvenile rainbow trout after administration by three different methods-bath, oral intubation into the stomach, and phage-coated feed-with special emphasis on the oral route of delivery. Phages could be detected in all the organs investigated (intestine, spleen, brain, and kidney) 0.5 h postadministration, reaching concentrations as high as ∼10(5) PFU mg intestine(-1) and ∼10(3) PFU mg spleen(-1) within the first 24 h following the bath and ∼10(7) PFU mg intestine(-1) and ∼10(4) PFU mg spleen(-1) within the first 24 h following oral intubation. The phages were most persistent in the organs for the first 24 h and then decreased exponentially; no phages were detected after 83 h in the organs investigated. Phage administration via feed resulted in the detection of phages in the intestine, spleen, and kidney 1 h after feeding. Average concentrations of ∼10(4) PFU mg intestine(-1) and ∼10(1) PFU mg spleen(-1) were found throughout the experimental period (200 h) following continuous delivery of phages with feed. These experiments clearly demonstrate the ability of the phages to survive passage through the fish stomach and to penetrate the intestinal barrier and enter the circulatory system after oral delivery, although the quantity of phages found in the spleen was 100- to 1,000-fold lower than that in the intestine. It was also shown that phages could tolerate long periods of desiccation on the feed pellets, with 60

Antimicrobial Susceptibility of Flavobacterium psychrophilum from Chilean Salmon Farms and their Epidemiological Cut-off Values using Agar Dilution and Disk Diffusion Methods

Directory of Open Access Journals (Sweden)

Claudio D Miranda

2016-11-01

Full Text Available Flavobacterium psychrophilum is the most important bacterial pathogen for freshwater farmed salmonids in Chile. The aims of this study were to determine the susceptibility to antimicrobials used in fish farming of Chilean isolates and to calculate their epidemiological cut-off (COWT values. A number of 125 Chilean isolates of F. psychrophilum were isolated from reared salmonids presenting clinical symptoms indicative of flavobacteriosis and their identities were confirmed by 16S rRNA polymerase chain reaction. Susceptibility to antibacterials was tested on diluted Mueller-Hinton by using an agar dilution MIC method and a disk diffusion method. The COWT values calculated by Normalised Resistance Interpretation (NRI analysis allow isolates to be categorized either as wild-type fully susceptible (WT or as manifesting reduced susceptibility (NWT. When MIC data was used, NRI analysis calculated a COWT of ≤ 0.125 μg mL-1, ≤ 2 μg mL-1 and ≤ 0.5 μg mL-1 for amoxicillin, florfenicol and oxytetracycline, respectively. For the quinolones, the COWT were ≤1 μg mL-1, ≤ 0.5 μg mL-1 and ≤ 0.125 μg mL-1 for oxolinic acid, flumequine and enrofloxacin respectively. The disc diffusion data sets obtained in this work were extremely diverse and were spread over a wide range. For the quinolones there was a close agreement between the frequencies of NWT isolates calculated using MIC and disc data. For oxolinic acid, flumequine and enrofloxacin the frequencies were 45, 39 and 38% using MIC data, and 42, 41 and 44%, when disc data were used. There was less agreement with the other antimicrobials, because NWT frequencies obtained using MIC and disc data respectively, were 24% and 10% for amoxicillin, 8% and 2% for florfenicol and 70% and 64% for oxytetracycline. Considering that the MIC data was more precise than the disc diffusion data, MIC determination would be the preferred method for susceptibility testing for this species and the NWT frequencies

Epidemiology and Control of Infectious Diseases of Salmonids in the Columbia River Basin, 1986 Annual Report.

Energy Technology Data Exchange (ETDEWEB)

Fryer, John L.

1986-12-01

The Department of Microbiology at Oregon State University with funding from the Bonneville Power Administration conducted a study relating to the epidemiology and control of three fish diseases of salmonids in the Columbia River Basin. These three diseases were ceratomyxosis caused by the myxosporidan parasite Ceratomyxa Shasta, bacterial kidney disease, the causative agent Renibacterium salmoninarum, and infectious hematopoietic necrosis, caused by a rhabdovirus. Each of these diseases is highly destructive and difficult or impossible to treat with antimicrobial agents. The geographic range of the infectious stage of C. Shasta has been extended to include the Snake River to the Oxbow and Hells Canyon Dams. These are the farthest upriver sites tested. Infections of ceratomyxosis were also initiated in the east fork of the Lewis River and in the Washougal River in Washington. Laboratory studies with this parasite failed to indicate that tubeficids are required in its life cycle. Bacterial kidney disease has been demonstrated in all life stages of salmonids: in the eggs, fry, smolts, juveniles and adults in the ocean, and in fish returning to fresh water. Monoclonal antibodies produced against R. salmoninarum demonstrated antigenic differences among isolates of the bacterium. Monoclonal antibodies also showed antigens of R. salmoninarum which are similar to those of a wide variety of gram positive and gram negative bacteria. A demonstration project at Round Butte Hatchery showed U V treatment to be an effective method for reducing the microbial population of the water supply and could reduce risks of IHNV. Tangential flow filtration was used successfully to concentrate IHNV from environmental water. At Round Butte Hatchery the carrier rate of IHNV in adults was very low and there was no subsequent mortality resulting from IHN in juveniles.

Disinfection of bacterial biofilms in pilot-scale cooling tower systems.

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Liu, Yang; Zhang, Wei; Sileika, Tadas; Warta, Richard; Cianciotto, Nicholas P; Packman, Aaron I

2011-04-01

The impact of continuous chlorination and periodic glutaraldehyde treatment on planktonic and biofilm microbial communities was evaluated in pilot-scale cooling towers operated continuously for 3 months. The system was operated at a flow rate of 10,080 l day(-1). Experiments were performed with a well-defined microbial consortium containing three heterotrophic bacteria: Pseudomonas aeruginosa, Klebsiella pneumoniae and Flavobacterium sp. The persistence of each species was monitored in the recirculating cooling water loop and in biofilms on steel and PVC coupons in the cooling tower basin. The observed bacterial colonization in cooling towers did not follow trends in growth rates observed under batch conditions and, instead, reflected differences in the ability of each organism to remain attached and form biofilms under the high-through flow conditions in cooling towers. Flavobacterium was the dominant organism in the community, while P. aeruginosa and K. pneumoniae did not attach well to either PVC or steel coupons in cooling towers and were not able to persist in biofilms. As a result, the much greater ability of Flavobacterium to adhere to surfaces protected it from disinfection, whereas P. aeruginosa and K. pneumoniae were subject to rapid disinfection in the planktonic state.

Sensitivity of some nitrogen fixers and the target pest Fusarium oxysporum to fungicide thiram.

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Osman, Awad G; Sherif, Ashraf M; Elhussein, Adil A; Mohamed, Afrah T

2012-03-01

This study was carried out to investigate the toxic effects of the fungicide thiram (TMTD) against five nitrogen fixers and the thiram target pest Fusarium oxysporum under laboratory conditions. Nitrogen fixing bacteria Falvobacterium showed the highest values of LD(50) and proved to be the most resistant to the fungicide followed by Fusarium oxysporum, while Pseudomonas aurentiaca was the most affected microorganism. LD(50) values for these microorganisms were in 2-5 orders of magnitude lower in comparison with LD(50) value for Fusarium oxysporum. Thiram was most toxic to Pseudomonas aurentiaca followed by Azospirillum. The lowest toxicity index was recorded for Fusarium oxysporum and Flavobacterium. The slope of the curve for Azomonas, Fusarium oxysporum and Flavobacterium is more steep than that of the other curves, suggesting that even a slight increase of the dose of the fungicide can cause a very strong negative effect. Thiram was more selective to Pseudomonas aurentiaca followed by Azospirillum, Rhizobium meliloti and Azomonas. The lowest selectivity index of the fungicide was recorded for Falvobacterium followed by Fusarium oxysporum. The highest safety coefficient of the fungicide was assigned for Flavobacterium, while Pseudomonas aurentiaca showed the lowest value.

Molecular diagnosis of infectious hematopoietic necrosis and viral hemorrhagic septicemia

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Winton, James R.; Einer-Jensen, Katja

2002-01-01

The fish rhabdoviruses, infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV), cause extensive losses among salmon and trout in several areas of the world (Bootland and Leong, 1999; Smail, 1999; Wolf, 1988). Historically, IHNV was endemic among wild anadromous salmonids in the western portion of North America, but the virus has spread to stocks of cultured rainbow trout (Oncorhynchus mykiss) in the United States, Asia and Western Europe, probably as a result of the movement of infected fish or eggs (Winton, 1991). Prior to 1989, VHSV was thought to be largely restricted to freshwater fishes in Western Europe (Wolf, 1988); however, in the last decade, VHSV has been isolated from an increasing number of free-living marine fish species in the North Pacific and North Atlantic Oceans (Dixon et al., 1997; Dixon, 1999; Kent et al., 1998; Meyers and Winton, 1995; Meyers et al., 1999; Mortensen et al., 1999; Smail; 2000, Takano et al., 2000). These findings have lead to the conclusion that both viruses are principally endemic among marine or anadromous fish species, but have established themselves in freshwater among cultured salmonids where their effects are most frequently observed.

Replication and shedding kinetics of infectious hematopoietic necrosis virus in juvenile rainbow trout

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Wargo, Andrew R.; Scott, Robert J.; Kerr, Benjamin; Kurath, Gael

2017-01-01

Viral replication and shedding are key components of transmission and fitness, the kinetics of which are heavily dependent on virus, host, and environmental factors. To date, no studies have quantified the shedding kinetics of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss), or how they are associated with replication, making it difficult to ascertain the transmission dynamics of this pathogen of high agricultural and conservation importance. Here, the replication and shedding kinetics of two M genogroup IHNV genotypes were examined in their naturally co-evolved rainbow trout host. Within host virus replication began rapidly, approaching maximum values by day 3 post-infection, after which viral load was maintained or gradually dropped through day 7. Host innate immune response measured as stimulation of Mx-1 gene expression generally followed within host viral loads. Shedding also began very quickly and peaked within 2 days, defining a generally uniform early peak period of shedding from 1 to 4 days after exposure to virus. This was followed by a post-peak period where shedding declined, such that the majority of fish were no longer shedding by day 12 post-infection. Despite similar kinetics, the average shedding rate over the course of infection was significantly lower in mixed compared to single genotype infections, suggesting a competition effect, however, this did not significantly impact the total amount of virus shed. The data also indicated that the duration of shedding, rather than peak amount of virus shed, was correlated with fish mortality. Generally, the majority of virus produced during infection appeared to be shed into the environment rather than maintained in the host, although there was more retention of within host virus during the post-peak period. Viral virulence was correlated with shedding, such that the more virulent of the two genotypes shed more total virus. This fundamental understanding of IHNV

Characterization of the interferon genes in homozygous rainbow trout reveals two novel genes, alternate splicing and differential regulation of duplicated genes

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Purcell, M.K.; Laing, K.J.; Woodson, J.C.; Thorgaard, G.H.; Hansen, J.D.

2009-01-01

The genes encoding the type I and type II interferons (IFNs) have previously been identified in rainbow trout and their proteins partially characterized. These previous studies reported a single type II IFN (rtIFN-??) and three rainbow trout type I IFN genes that are classified into either group I (rtIFN1, rtIFN2) or group II (rtIFN3). In this present study, we report the identification of a novel IFN-?? gene (rtIFN-??2) and a novel type I group II IFN (rtIFN4) in homozygous rainbow trout and predict that additional IFN genes or pseudogenes exist in the rainbow trout genome. Additionally, we provide evidence that short and long forms of rtIFN1 are actively and differentially transcribed in homozygous trout, and likely arose due to alternate splicing of the first exon. Quantitative reverse transcriptase PCR (qRT-PCR) assays were developed to systematically profile all of the rainbow trout IFN transcripts, with high specificity at an individual gene level, in na??ve fish and after stimulation with virus or viral-related molecules. Cloned PCR products were used to ensure the specificity of the qRT-PCR assays and as absolute standards to assess transcript abundance of each gene. All IFN genes were modulated in response to Infectious hematopoietic necrosis virus (IHNV), a DNA vaccine based on the IHNV glycoprotein, and poly I:C. The most inducible of the type I IFN genes, by all stimuli tested, were rtIFN3 and the short transcript form of rtIFN1. Gene expression of rtIFN-??1 and rtIFN-??2 was highly up-regulated by IHNV infection and DNA vaccination but rtIFN-??2 was induced to a greater magnitude. The specificity of the qRT-PCR assays reported here will be useful for future studies aimed at identifying which cells produce IFNs at early time points after infection. ?? 2008 Elsevier Ltd.

Studies on antagonistic marine streptomycetes

Digital Repository Service at National Institute of Oceanography (India)

Chandramohan, D.; Nair, S.

three strains inhibited all the test cultures. In addition to the above test cultures marine bacteria (Vibrio sp., Aeromonas spp., Flavobacterium spp., Bacillus sp. and Micrococcus sp.) resistant to few known antibiotics (tetracycline, penicillin...

Role of bacterial adhesion in the microbial ecology of biofilms in cooling tower systems.

Science.gov (United States)

Liu, Yang; Zhang, Wei; Sileika, Tadas; Warta, Richard; Cianciotto, Nicholas P; Packman, Aaron

2009-01-01

The fate of the three heterotrophic biofilm forming bacteria, Pseudomonas aeruginosa, Klebsiella pneumoniae and Flavobacterium sp. in pilot scale cooling towers was evaluated both by observing the persistence of each species in the recirculating water and the formation of biofilms on steel coupons placed in each cooling tower water reservoir. Two different cooling tower experiments were performed: a short-term study (6 days) to observe the initial bacterial colonization of the cooling tower, and a long-term study (3 months) to observe the ecological dynamics with repeated introduction of the test strains. An additional set of batch experiments (6 days) was carried out to evaluate the adhesion of each strain to steel surfaces under similar conditions to those found in the cooling tower experiments. Substantial differences were observed in the microbial communities that developed in the batch systems and cooling towers. P. aeruginosa showed a low degree of adherence to steel surfaces both in batch and in the cooling towers, but grew much faster than K. pneumoniae and Flavobacterium in mixed-species biofilms and ultimately became the dominant organism in the closed batch systems. However, the low degree of adherence caused P. aeruginosa to be rapidly washed out of the open cooling tower systems, and Flavobacterium became the dominant microorganism in the cooling towers in both the short-term and long-term experiments. These results indicate that adhesion, retention and growth on solid surfaces play important roles in the bacterial community that develops in cooling tower systems.

Aerobic Denitrification as an Innovative Method for In-Situ Biological Remediation of Contaminated Subsurface Sites

Science.gov (United States)

1989-01-01

roseus Bacillus stearothermophilus Micrococcus varians Paracoccus denitrificans Bacillus coagulans Paracoccus halodenitrificans Bacillus flrmus Bacillus ...Geodermatophilus Plesiomonas Arachnia Haemophilus Propionibacterium Arthrobacter Halobacteriua Proteus Bacillus Ifalocuccus Pseudoraonas Bacteroides...Mycobacterium peregrinum Nocardia opaca Chromobacterium violaceum Bacillus subtllis Nocardia atlantica Bacillus licheniformis Flavobacterium

Dicty_cDB: Contig-U03062-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 1.2 11 ( CP000685 ) Flavobacterium johnsoniae UW101, complete genome. 36 1.3 17 ( FG292300 ) 1108457713646 New World... Screwworm Larvae 9387 EST... 36 1.3 2 ( FG293205 ) 1108770666478 New World Screwworm Larvae 9387 ES

Reduction of rainbow trout spleen size by splenectomy does not alter resistance against bacterial cold water disease

Science.gov (United States)

In lower vertebrates, the contribution of the spleen to anti-bacterial immunity is poorly understood. Researchers have previously reported a phenotypic and genetic correlation between resistance to Flavobacterium psychrophilum, the causative agent of bacterial cold water disease (BCWD) and spleen so...

The stress hormone cortisol: a (co)regulator of biofilm formation in Flavobacterum columnare?

Science.gov (United States)

Previously, we demonstrated a direct effect of cortisol on Flavobacterium columnare, a notorious fish pathogenic bacterium, engendering a new perspective to bacteria-host communication in aquaculture. As stressed fish harbour increased cortisol levels in the skin and gill mucus, highly virulent F. c...

Impacts of parasite infection on columnaris disease of tilapia

Science.gov (United States)

There is no information available on whether parasite infection will increase the susceptibility of tilapia to Flavobacterium columnare and whether parasite treatment could improve fish survival after F. columnare exposure. Two trials were conducted to evaluate 1) the susceptibility of hybrid tilapi...

Iron acquisition mechanisms of Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Møller, Jeannette Dan; Ellis, A.E.; Barnes, A.C.

2005-01-01

strains and was lacking from negative strains. While a weak catechol reaction was detectable in CAS+ culture supernatants, the CAS reaction was, to some extent, heat sensitive, questioning whether the positive reaction was caused only by siderophores. The ability to grow in vitro under iron......-restricted conditions did not correlate with the CAS reactivity, as growth of both CAS+ and CAS- strains was similarly impaired under iron restriction induced by 2,2 dipyridyl. Suppressed growth under these conditions was restored by addition of FeCl3, haemoglobin and transferrin for both CAS+ and CAS- strains...

Nitrous oxide reductase genes (nosZ) of denitrifying populations in soil and the earthworm gut are phylogenetically similar

DEFF Research Database (Denmark)

Horn, Marcus A.; Drake, Harold L.; Schramm, Andreas

2006-01-01

-derived sequences, or were related to N2O reductases of the genera Bradyrhizobium, Brucella, Dechloromonas, Flavobacterium, Pseudomonas, Ralstonia, and Sinorhizobium. Although the numbers of estimators for genotype richness of sequence data from the gut were higher than those of soil, only one gut-derived nos...

Rapid detection and identification of viral and bacterial fish pathogens using a DNA array‐based multiplex assay

DEFF Research Database (Denmark)

Lievens, B.; Frans, I.; Heusdens, C.

2011-01-01

for the simultaneous detection and identification of all cyprinid herpesviruses (CyHV‐1, CyHV‐2 and CyHV‐3) and some of the most important fish pathogenic Flavobacterium species, including F. branchiophilum, F. columnare and F. psychrophilum. For virus identification, the DNA polymerase and helicase genes were...

Twenty-one genome sequences from Pseudomonas species and 19 genome sequences from diverse bacteria isolated from the rhizosphere and endosphere of Populus deltoides.

Science.gov (United States)

Brown, Steven D; Utturkar, Sagar M; Klingeman, Dawn M; Johnson, Courtney M; Martin, Stanton L; Land, Miriam L; Lu, Tse-Yuan S; Schadt, Christopher W; Doktycz, Mitchel J; Pelletier, Dale A

2012-11-01

To aid in the investigation of the Populus deltoides microbiome, we generated draft genome sequences for 21 Pseudomonas strains and 19 other diverse bacteria isolated from Populus deltoides roots. Genome sequences for isolates similar to Acidovorax, Bradyrhizobium, Brevibacillus, Caulobacter, Chryseobacterium, Flavobacterium, Herbaspirillum, Novosphingobium, Pantoea, Phyllobacterium, Polaromonas, Rhizobium, Sphingobium, and Variovorax were generated.

Twenty-One Genome Sequences from Pseudomonas Species and 19 Genome Sequences from Diverse Bacteria Isolated from the Rhizosphere and Endosphere of Populus deltoides

Energy Technology Data Exchange (ETDEWEB)

Brown, Steven D [ORNL; Utturkar, Sagar M [ORNL; Klingeman, Dawn Marie [ORNL; Johnson, Courtney M [ORNL; Martin, Stanton [ORNL; Land, Miriam L [ORNL; Lu, Tse-Yuan [ORNL; Schadt, Christopher Warren [ORNL; Doktycz, Mitchel John [ORNL; Pelletier, Dale A [ORNL

2012-01-01

To aid in the investigation of the Populus deltoides microbiome we generated draft genome sequences for twenty one Pseudomonas and twenty one other diverse bacteria isolated from Populus deltoides roots. Genome sequences for isolates similar to Acidovorax, Bradyrhizobium, Brevibacillus, Burkholderia, Caulobacter, Chryseobacterium, Flavobacterium, Herbaspirillum, Novosphingobium, Pantoea, Phyllobacterium, Polaromonas, Rhizobium, Sphingobium and Variovorax were generated.

Flavobacterium psychrophilum - Experimental challenge and immune response

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi

the immune system of the fry is not fully developed. Theoretically, the infection pressure could be subdued by vaccinating larger fish, but no commercial vaccine is yet available. Diagnostic methods are well described and the disease is treated with antibiotics. To prevent disease outbreaks and subsequent......-time PCR (RT-PCR) was used to examine the immune response in the head kidney during the first eight days after infection, and enzyme-linked immunosorbent assay (ELISA) was used to evaluate the production of antibodies 50 days post-exposure. A pro-inflammatory response was observed in both groups infected...... of edemas, but in both cases the tissue was regenerating after 192 hours. However, when the fish had been exposed to both H2O2 and F. psychrophilum, the damage was still evident at this time point. The relative pathogen load measured as 16S rRNA was highest at the first sampling and decreased steadily...

The effect of prolonged flooding of an oil deposit on the special composition and the activity of hydrocarbon-oxidizing microflora

Energy Technology Data Exchange (ETDEWEB)

Berdichevskaya, M V

1982-07-01

The special composition of hydrocarbon-oxidizing bacteria was studied in terrigenous and carbonate oil-bearing strata from several deposits of the Permian Cis-Ural region. We isolated 43 strains and assigned them to the following genera: Mycobacterium, Micrococcus, Brevibacterium, Corynebacterium, Flavobacterium, Achromobacter and Pseudomonas. The special composition of the hydrocarbon-oxidizing microflora was shown to depend on the flooding of an oil stratum, as a result of which the ecological environment in a deposit changed. Gram-positive coryneform bacteria were found in stratal salinized waters and in diluted stratal waters. Gram-negative hydrocarbon-oxidizing bacteria were isolated from pumped-in river waters and from stratal waters diluted by 70-100% as the result of flooding. The metabolic activity of Corynebacterium fascians (2 strains), Mycobacterium rubrum (1 strain), Pseudomonas mira (1 strain) and Flavobacterium perigrinum (1 strain) was assayed in stratal waters with different concentrations of salts. The coryneform hydrocarbon-oxidizing bacteria were shown to be very halotolerant as the result of adaptation; that is why the incidence of these microorganisms is very great in highly mineralized stratal water of oil deposits.

Pathogen Screening of Naturally Produced Yakima River Spring Chinook Smolts; Yakima/Klickitat Fisheries Project Monitoring and Evaluation Report 6 of 7, 2003-2004 Annual Report.

Energy Technology Data Exchange (ETDEWEB)

Thomas, Joan B. (Washington Department of Fish and Wildlife, Olympia, WA)

2004-05-01

In 1999 the Cle Elum Hatchery began releasing spring chinook salmon smolts into the upper Yakima River to increase natural production. Part of the evaluation of this program is to monitor whether introduction of hatchery produced smolts would impact the prevalence of specific pathogens in the naturally produced spring chinook smolts. Increases in prevalence of any of these pathogens could negatively impact the survival of these fish. In 1998 and 2000 through 2003 naturally produced smolts were collected for monitoring at the Chandler smolt collection facility on the lower Yakima River. Smolts were collected from mid to late outmigration, with a target of 200 fish each year. The pathogens monitored were infectious hematopoeitic necrosis virus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, Flavobacterium psychrophilum, Flavobacterium columnare, Aeromonas salmonicida, Yersinia ruckeri, Edwardsiella ictaluri, Renibacterium salmoninarum and Myxobolus cerebralis. To date, only the bacterial pathogens have been detected and prevalences have been low. Prevalences have varied each year and these changes are attributed to normal fluctuation of prevalence. All of the pathogens detected are widely distributed in Washington State.

Glyphosate-Degrading Microorganisms from Industrial Activated Sludge

OpenAIRE

Balthazor, Terry M.; Hallas, Laurence E.

1986-01-01

A plating medium was developed to isolate N-phosphonomethylglycine (glyphosate)-degrading microorganisms, with glyphosate as the sole phosphorus source. Two industrial biosystems treating glyphosate wastes contained elevated microbial counts on the medium. One purified isolate metabolized glyphosate to aminomethylphosphonic acid, mineralizing this accumulating intermediate during log growth. This microorganism has been identified as a Flavobacterium species.

Rapid Diagnosis of IHN Virus Infection in Salmon and Steelhead Trout, Final Report.

Energy Technology Data Exchange (ETDEWEB)

Leong, JoAnn Ching

1984-12-01

The main objective for this study was the development of a rapid diagnostic method for IHN virus in fish tissue samples. The rationale for developing new techniques for diagnosing IHNV infection was that present methods were time consuming and dependent on virus neutralization by specific antisera, a reagent that was not readily available or reliable. Fish pathologists required a rapid detection method which was sensitive enough to detect virus strain differences so that they could provide data for effective management decisions in controlling the spread of IHNV. Bonneville Power Administration's (BPA) role in efforts in fish diseases and more generically the protection, mitigation, and enhancement of Columbia River salmon and steelhead populations, is mandated by Congress through the Pacific Northwest Electric Power Planning and Conservation Act (Regional Act), Pub. L. 96-501. Section 4 (h) of the Regional Act directs the Northwest Power Planning Council to develop a Fish and Wildlife Program. BPA's Administrator is authorized in Section 4 (h) (10) (A) to ''use funds and the authorities available to the extent affected by the development and operation of any hydroelectric project of the Columbia River and its tributaries''. The fund is to be used to implement measures that are consistent with the Council's Fish and Wildlife Program. The research detailed in this final report is consistent with these objectives. This final report has been prepared as part of BPA's policy to encourage the preservation and dissemination of research results by publication in scientific journals.

Sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral hemorrhagic septicemia virus, a fish rhabdovirus

Science.gov (United States)

Benmansour, A.; Bascuro, B.; Monnier, A.F.; Vende, P.; Winton, J.R.; de Kinkelin, P.

1997-01-01

To evaluate the genetic diversity of viral haemorrhagic septicaemia virus (VHSV), the sequence of the glycoprotein genes (G) of 11 North American and European isolates were determined. Comparison with the G protein of representative members of the family Rhabdoviridae suggested that VHSV was a different virus species from infectious haemorrhagic necrosis virus (IHNV) and Hirame rhabdovirus (HIRRV). At a higher taxonomic level, VHSV, IHNV and HIRRV formed a group which was genetically closest to the genus Lyssavirus. Compared with each other, the G genes of VHSV displayed a dissimilar overall genetic diversity which correlated with differences in geographical origin. The multiple sequence alignment of the complete G protein, showed that the divergent positions were not uniformly distributed along the sequence. A central region (amino acid position 245-300) accumulated substitutions and appeared to be highly variable. The genetic heterogeneity within a single isolate was high, with an apparent internal mutation frequency of 1.2 x 10(-3) per nucleotide site, attesting the quasispecies nature of the viral population. The phylogeny separated VHSV strains according to the major geographical area of isolation: genotype I for continental Europe, genotype II for the British Isles, and genotype III for North America. Isolates from continental Europe exhibited the highest genetic variability, with sub-groups correlated partially with the serological classification. Neither neutralizing polyclonal sera, nor monoclonal antibodies, were able to discriminate between the genotypes. The overall structure of the phylogenetic tree suggests that VHSV genetic diversity and evolution fit within the model of random change and positive selection operating on quasispecies.

Analysis of host genetic diversity and viral entry as sources of between-host variation in viral load

Science.gov (United States)

Wargo, Andrew R.; Kell, Alison M.; Scott, Robert J.; Thorgaard, Gary H.; Kurath, Gael

2012-01-01

Little is known about the factors that drive the high levels of between-host variation in pathogen burden that are frequently observed in viral infections. Here, two factors thought to impact viral load variability, host genetic diversity and stochastic processes linked with viral entry into the host, were examined. This work was conducted with the aquatic vertebrate virus, Infectious hematopoietic necrosis virus (IHNV), in its natural host, rainbow trout. It was found that in controlled in vivo infections of IHNV, a suggestive trend of reduced between-fish viral load variation was observed in a clonal population of isogenic trout compared to a genetically diverse population of out-bred trout. However, this trend was not statistically significant for any of the four viral genotypes examined, and high levels of fish-to-fish variation persisted even in the isogenic trout population. A decrease in fish-to-fish viral load variation was also observed in virus injection challenges that bypassed the host entry step, compared to fish exposed to the virus through the natural water-borne immersion route of infection. This trend was significant for three of the four virus genotypes examined and suggests host entry may play a role in viral load variability. However, high levels of viral load variation also remained in the injection challenges. Together, these results indicate that although host genetic diversity and viral entry may play some role in between-fish viral load variation, they are not major factors. Other biological and non-biological parameters that may influence viral load variation are discussed.

Bacterial Community Associated with Fish and Water from Congonhas River, Sertaneja, Paraná, Brazil

Directory of Open Access Journals (Sweden)

José Américo de Sousa

2001-12-01

Full Text Available A bacteriological study was conducted on fish and water from Congonhas River, Sertaneja (22º58’ S; 50º58’ W, Paraná State, Brazil. From 44% of the analysed fish, bacteria belonging to Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Micrococcus, Bacillus and Lactobacillus were isolated. The group most frequently isolated from fish was Aeromonas. In the water, the bacterial groups detected were Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Bacillus and Flavobacterium, from which Flavobacterium and Acinetobacter were the most abundant. The numbers of Colony Forming Units per millilitre of water varied from 3.1x10² to 1.0 x 10³. Although a clear pattern was not detected in the susceptibilities/resistances of the isolated strains to nine antimicrobial substances, Gram negative aerobic bacteria were more resistant than the other strains. A simultaneous resistance to furazolidone, oxolinic acid and norfloxacin, particularly in the bacteria isolated from fish, as well as in the aerobic strains isolated from water was observed. The antimicrobial substances to which less resistances were found were oxytetracycline in the strains isolated from water, and trimethoprim-sulphamethoxazole, oxytetracycline and chloramphenicol in those isolated from fish.Foi realizado um estudo da comunidade bacteriana de peixes e da água do rio Congonhas, próximo à sua foz no rio Tibagi, município de Sertaneja, Paraná, Brasil. De 44% dos peixes analisados, foram isoladas estirpes de Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Micrococcus, Bacillus e Lactobacillus. Destas, a mais abundante foi Aeromonas. Na água do rio Congonhas foram detectados os grupos Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Bacillus e Flavobacterium, dos quais os predominantes foram Flavobacterium e Acinetobacter. Os números de unidades formadoras de colônias por mililitro de água variaram entre 3,1x10² e 1,0x10³. Embora não tenha

Occurrence of Antimicrobial Resistance in Fish-Pathogenic and Environmental Bacteria Associated with Four Danish Rainbow Trout Farms

DEFF Research Database (Denmark)

Schmidt, Anja S.; Bruun, Morten Sichlau; Dalsgaard, Inger

2000-01-01

in fish, water, and sediment samples, two major fish pathogens (88 Flavobacterium psychrophilum isolates and 134 Yersinia ruckeri isolates) and 313 motile Aeromonas isolates, representing a group of ubiquitous aquatic bacteria, were isolated from the same samples. MICs were obtained applying...... flavobacteria and aeromonads, thus indicating a substantial impact of fish farming on several groups of bacteria associated with aquacultural environments....

EFFECT OF REFINED PETROLEUM PRODUCTS CONTAMINATION ON BACTERIAL POPULATION AND PHYSICOCHEMICAL CHARACTERISTICS OF CULTIVATED AGRICULTURAL SOIL

Directory of Open Access Journals (Sweden)

Adewale Sogo Olalemi

2012-10-01

Full Text Available An investigation into the effect of refined petroleum products contamination on bacterial population and physicochemical characteristics of cultivated agricultural soil was carried out. The soil samples obtained from the Teaching and Research Farm, Obakekere, Federal University of Technology, Akure, Ondo State were contaminated with varying volumes of petrol, diesel and kerosene. The results revealed higher bacterial populations in uncontaminated soils than contaminated soils. The counts of bacteria ranged from 3.0 × 105 to 5.0 × 105 cfu/g in uncontaminated soils and 1.0 × 105 to 3.0 × 105 cfu/g in contaminated soils. The isolated bacteria were identified as Bacillus subtilis, Flavobacterium lutescens, Micrococcus luteus, Corynebacterium variabilis, Pseudomonas fluorescens. The contamination had no significant effect on pH, potassium, sodium, organic carbon and nitrogen content of the soils, while the moisture, calcium, phosphorus and magnesium content of the contaminated soils were significantly different (P < 0.05 compared with the uncontaminated soils. The ability of Bacillus subtilis, Flavobacterium lutescens, Micrococcus luteus, and Pseudomonas fluorescens to utilize the refined petroleum products suggest that these bacteria had potential to bioremediate petroleum contaminated soils.

21 CFR 866.3270 - Flavobacterium spp. serological reagents.

Science.gov (United States)

2010-04-01

... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3270.... from cultured isolates derived from clinical specimens. The identification aids in the diagnosis of...

Rainbow trout surviving infections of viral haemorrhagic septicemia virus (VHSV) show lasting antibodies to recombinant G protein fragments

DEFF Research Database (Denmark)

Encinas, P.; Gomez-Casado, E.; Grandes, Fregeneda

2011-01-01

sera from trout infected with the homologous VHSV isolate but also with the VHSV-DK-201433 heterologous isolate, which had 13 amino acid changes. Sera from healthy trout and/or from trout surviving infectious haematopoietic necrosis virus (IHNV) infection, were used to calculate cut-off absorbances...... refinements of the frg-ELISA could allow detection of anti-VHSV trout Abs in natural outbreaks caused by different heterologous VHSV isolates. The homologous frg-ELISA method could be useful to follow G immunization attempts during vaccine development and/or to best understand the fish Ab response during VHSV...... infections. The viral frgs approach might also be used with other fish species and/or viruses....

Effects of associated bacteria on the pathogenicity and reproduction of the insect-parasitic nematode Rhabditis blumi (Nematoda: Rhabditida).

Science.gov (United States)

Park, Hae Woong; Kim, Yong Ook; Ha, Jae-Seok; Youn, Sung Hun; Kim, Hyeong Hwan; Bilgrami, Anwar L; Shin, Chul Soo

2011-09-01

Three bacteria, Alcaligenes faecalis , Flavobacterium sp., and Providencia vermicola , were isolated from dauer juveniles of Rhabditis blumi . The pathogenic effects of the bacteria against 4th instar larvae of Galleria mellonella were investigated. Providencia vermicola and Flavobacterium sp. showed 100% mortality at 48 h after haemocoelic injection, whereas A. faecalis showed less than 30% mortality. Dauer juveniles showed 100% mortality against G. mellonella larvae, whereas axenic juveniles, which do not harbor associated bacteria, exhibited little mortality. All of the associated bacteria were used as a food source for nematode growth, and nematode yield differed with bacterial species. Among the bacterial species, P. vermicola was most valued for nematode yield, showing the highest yield of 5.2 × 10(4) nematodes/mL in the plate. In bacterial cocultures using two of the three associated bacteria, one kind stimulated the other. The highest total bacterial yield of 12.6 g/L was obtained when the inoculum ratio of P. vermicola to A. faecalis was 10:1. In air-lift bioreactors, the nematode growth rate increased with an increasing level of dissolved oxygen. The maximum nematode yield of 1.75 × 10(5) nematodes/mL was obtained at 192 h with an aeration rate of 6 vvm.

Comparison of infectious hematopoietic necrosis in natural and experimental infections of spawning salmonids by infectivity and immunohistochemistry

Science.gov (United States)

Yamamoto, T.; Arakawa, C.K.; Batts, W.N.; Winton, J.R.

1989-01-01

Infectious hematopoietic necrosis (IHN) continues to be a serious virus disease of salmonids with epizootics recorded in both wild and hatchery populations (Williams and Amend 1976; Carlisle et al 1979; Groberg and Fryer 1983; Saft and Pratt 1986; Traxler 1987; Follett et al 1987; Meyers et al 1988). While originally enzootic in western North America, the virus appears to be spreading further (Sano et al 1977; de Kinkelin et al 1987; Bovo et al 1987). In hatchery outbreaks occurring in regions where the virus is not enzootic, it is often possible to trace the virus to the importation of infected fingerlings or contaminated eggs. In regions where the virus is widespread among stocks of fish, the source of virus infection is more difficult to establish particularly in watersheds where there are anadromous salmonids. Although salmonid fish surviving infection as fry and returning from the ocean to spawn are considered to be parental carriers of IHNV, there is very little data to support this hypothesis. Amend (1975) in the study of rainbow trout reported that in a population surviving infection and assayed a few years later found that a few trout were carrying virus. This is the study often cited as evidence for the carrier status of returning salmonids. LaPatra et al (1987) presented data that indicated IHNV has been transmitted horizontally through water from adult chinook salmon (Oncorhynchus tshawytscha) to adult coho salmon (O. kisutch) at a hatchery in northern California. They suggested that horizontal transmission may be an important means for perpetuating IHN. However, the actual mechanisms for persistence and transmission of IHN among fish in a watershed is likely to be complex and involve multiple species and age groups as well as intermediate vectors and/or reservoirs.

The Military Efficacy of Individual Water Purification Filters.

Science.gov (United States)

1990-12-01

criteria. ’t *.;as a.Lso serv ea that the KPF units grew opportunistic Pseudomonas sp . on the oroduct water side of the filters, which could have a...small pathogenic protozoan cysts such as Cryptospor-dium parvum, Giardia lamblia, and Entamoeba coli, from water over the effective use life of the...of heterotrophic bacteria such as Pseudoronas sP , Flavobacterium Sp , and other potentially harmfui organisms inside of the units. Enteric viruses are

Xanthomonads and other yellow-pigmented, Xanthomonas-like bacteria associated with tomato seeds in Tanzania

DEFF Research Database (Denmark)

Mbega, Ernest Rashid; Wulff, Ednar Gadelha; Mabagala, R.B.

2012-01-01

of Xanthomonas campestris pv. malvacearum, were pathogenic on tomato and pepper plants. Strains identified by Biolog as Sphingomonas sanguinis and Sphingomonas terrae also incited black rot symptoms on pepper leaves. However, bacterial strains belonging to the genus Stenotrophomonas, Chryseobacterium, Pantoea...... and Flavobacterium were not pathogenic on tomato and pepper. Phylogenetic analysis showed that strains of the genus Xanthomonas are more closely related to Stenotrophomonas and Pantoea compared to the other bacterial genera found in tomato seeds....

A selective and differential medium for Vibrio harveyi.

OpenAIRE

Harris, L; Owens, L; Smith, S

1996-01-01

A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated...

Bacterial microflora isolated from the bark surface of poplars growing in areas where air pollution is very high

Directory of Open Access Journals (Sweden)

Krystyna Przybył

2015-01-01

Full Text Available In the autumn of 1976 bacteria of the genera Bacillus, Pseudomonas, Flavobacterium, Erwinia and Cellulomonas were isolated from the bark surface of poplars growing in protective belts around several industrial plants. It was found that the qualitative and quantitative composition of the surface bacterial microflora changes in dependence on the degree of resistance of the poplars to the action of the dust emitted by the industrial establishment and containing high amounts of heavy metals.

RESPUESTA ANTAGÓNICA DE MICROORGANISMOS AISLADOS DE LOS CULTIVOS DE MICROALGAS EN EL LABORATORIO DE CULTIVOS MARINOS

Directory of Open Access Journals (Sweden)

Carmen Ruíz Huamán

2013-10-01

Full Text Available Durante el 2012, el Laboratorio de Cultivos Marinos obtuvo bacterias aisladas de los cultivos de microalgas. Se trabajó 60 colonias de los cultivos. Se confrontó una a una las colonias aisladas del cultivo de microalgas con las bacterias patógenas tipo ATCC, estas fueron: Flavobacterium psychrophilum, Aeromona hydrophila, Lactococcus garviaceae y Vibrio herveyi. Solo una bacteria tuvo interacción antagónica ya que formó el halo de inhibición.

Antimicrobial resistance patterns in Danish isolates of Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Bruun, Morten Sichlau; Schmidt, A.S.; Madsen, Lone

2000-01-01

were tested and the resulting antibiograms were used to predict the theoretical therapeutic efficacy and to evaluate if resistance had changed as a course of time. Antimicrobial agents included in this investigation were oxolinic acid (OXA), amoxicillin (AMX), potentiated sulfadiazine, oxytetracycline...

Inter individual variations of the fish skin microbiota: host genetics basis of mutualism?

Science.gov (United States)

Boutin, Sébastien; Sauvage, Christopher; Bernatchez, Louis; Audet, Céline; Derome, Nicolas

2014-01-01

The commensal microbiota of fish skin is suspected to provide a protection against opportunist infections. The skin of fish harbors a complex and diverse microbiota that closely interacts with the surrounding water microbial communities. Up to now there is no clear evidence as to whether the host regulates the recruitment of environmental bacteria to build a specific skin microbiota. To address this question, we detected Quantitative Trait Loci (QTL) associated with the abundance of specific skin microbiota bacterial strains in brook charr (Salvelinus fontinalis), combining 16S RNA tagged-amplicon 454 pyrosequencing with genetic linkage analysis. Skin microbiota analysis revealed high inter-individual variation among 86 F2 fish progeny based upon the relative abundance of bacterial operational taxonomic units (OTUs). Out of those OTUs, the pathogenic strain Flavobacterium psychrophilum and the non-pathogenic strain Methylobacterium rhodesianum explained the majority of inter-individual distances. Furthermore, a strong negative correlation was found between Flavobacterium and Methylobacterium, suggesting a mutually competitive relationship. Finally, after considering a total of 266 markers, genetic linkage analysis highlighted three major QTL associated with the abundance of Lysobacter, Rheinheimera and Methylobacterium. All these three genera are known for their beneficial antibacterial activity. Overall, our results provide evidence that host genotype may regulate the abundance of specific genera among their surface microbiota. They also indicate that Lysobacter, Rheinheimera and Methylobacterium are potentially important genera in providing protection against pathogens.

Pathogen Screening of Naturally Produced Yakima River Spring Chinook Smolts; Yakima/Klickitat Fisheries Project Monitoring and Evaluation, 2004-2005 Annual Report.

Energy Technology Data Exchange (ETDEWEB)

Thomas, Joan B. (Washington Department of Fish and Wildlife, Olympia, WA)

2005-05-01

In the spring of 2004 naturally produced smolts outmigrating from the Yakima River Basin were collected for the sixth year of pathogen screening. This component of the evaluation is to monitor whether introduction of hatchery produced smolts would impact the prevalence of specific pathogens in the naturally produced spring chinook smolts. Increases in prevalence of any of these pathogens could negatively impact the survival of these fish. Since 1999 the Cle Elum Hatchery has been releasing spring chinook salmon smolts into the upper Yakima River to increase natural production. In 1998 and 2000 through 2004 naturally produced smolts were collected for monitoring at the Chandler smolt collection facility on the lower Yakima River. Smolts were collected from mid to late outmigration, with a target of 200 fish each year. The pathogens monitored were infectious hematopoeitic necrosis virus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, Flavobacterium psychrophilum, Flavobacterium columnare, Aeromonas salmonicida, Yersinia ruckeri, Edwardsiella ictaluri, Renibacterium salmoninarum and Myxobolus cerebralis. Of these pathogens, only R. salmoninarum was detected in very low levels in the naturally produced smolts outmigrating in 2004. To date, only bacterial pathogens have been detected and prevalences have been low. There have been small variations each year and these changes are attributed to normal fluctuations in prevalence. All of the pathogens detected are widely distributed in Washington State.

Immunity induced shortly after DNA vaccination of rainbow trout against rhabdoviruses protects against heterologous virus but not against bacterial pathogens

DEFF Research Database (Denmark)

Lorenzen, Niels; Lorenzen, Ellen; Einer-Jensen, Katja

2002-01-01

whereas no increased survival was found upon challenge with bacterial pathogens. Within two months after vaccination, the cross-protection disappeared while the specific immunity to homologous virus remained high. The early immunity induced by the DNA vaccines thus appeared to involve short-lived non......It was recently reported that DNA vaccination of rainbow trout fingerlings against viral hemorrhagic septicaemia virus (VHSV) induced protection within 8 days after intramuscular injection of plasmid DNA. In order to analyse the specificity of this early immunity, fish were vaccinated with plasmid...... DNA encoding the VHSV or the infectious haematopoietic necrosis virus (IHNV) glycoprotein genes and later challenged with homologous or heterologous pathogens. Challenge experiments revealed that immunity established shortly after vaccination was cross-protective between the two viral pathogens...

Bioremediation of soil with diesel Through the use of autochthonous microorganisms

International Nuclear Information System (INIS)

Arrieta Ramirez, Olga Maria; Rivera Rivera, Angela Patricia; Arias Marin, Lida; Rojano, Benjamin Alberto; Ruiz, Orlando; Cardona Gallo, Santiago Alonso

2012-01-01

In this study was isolated and characterized biochemical and molecular a bacterial consortium able to degrade hydrocarbons several, comprised of the following genres: Enterobacter sp, Bacillus sp, Staphylococcus aureus, Sanguibacter soli, Arthrobacter spy Flavobacterium sp, from soil contaminated with diesel fuel in a laboratory scale, and treated with two technologies for bioremediation: natural attenuation and biostimulation. We obtained a reduction in the concentration of Total Petroleum Hydrocarbons (TPH) in a period of 4 months was 36,86% for natural attenuation and 50,99% for biostimulation.

Molecular Structure of Endotoxins from Gram-negative Marine Bacteria: An Update

Directory of Open Access Journals (Sweden)

Antonio Molinaro

2007-09-01

Full Text Available Marine bacteria are microrganisms that have adapted, through millions of years, to survival in environments often characterized by one or more extreme physical or chemical parameters, namely pressure, temperature and salinity. The main interest in the research on marine bacteria is due to their ability to produce several biologically active molecules, such as antibiotics, toxins and antitoxins, antitumor and antimicrobial agents. Nonetheless, lipopolysaccharides (LPSs, or their portions, from Gram-negative marine bacteria, have often shown low virulence, and represent potential candidates in the development of drugs to prevent septic shock. Besides, the molecular architecture of such molecules is related to the possibility of thriving in marine habitats, shielding the cell from the disrupting action of natural stress factors. Over the last few years, the depiction of a variety of structures of lipids A, core oligosaccharides and O-specific polysaccharides from LPSs of marine microrganisms has been given. In particular, here we will examine the most recently encountered structures for bacteria belonging to the genera Shewanella, Pseudoalteromonas and Alteromonas, of the γ-Proteobacteria phylum, and to the genera Flavobacterium, Cellulophaga, Arenibacter and Chryseobacterium, of the Cytophaga- Flavobacterium-Bacteroides phylum. Particular attention will be paid to the chemical features expressed by these structures (characteristic monosaccharides, non-glycidic appendages, phosphate groups, to the typifying traits of LPSs from marine bacteria and to the possible correlation existing between such features and the adaptation, over years, of bacteria to marine environments.

Shifts of microbial communities of wheat (Triticum aestivum L.) cultivation in a closed artificial ecosystem.

Science.gov (United States)

Qin, Youcai; Fu, Yuming; Dong, Chen; Jia, Nannan; Liu, Hong

2016-05-01

The microbial communities of plant ecosystems are in relation to plant growing environment, but the alteration in biodiversity of rhizosphere and phyllosphere microbial communities in closed and controlled environments is unknown. The purpose of this study is to analyze the change regularity of microbial communities with wheat plants dependent-cultivated in a closed artificial ecosystem. The microbial community structures in closed-environment treatment plants were investigated by a culture-dependent approach, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), and Illumina Miseq high-throughput sequencing. The results indicated that the number of microbes decreased along with time, and the magnitude of bacteria, fungi, and actinomycetes were 10(7)-10(8), 10(5), and 10(3)-10(4) CFU/g (dry weight), respectively. The analysis of PCR-DGGE and Illumina Miseq revealed that the wheat leaf surface and near-root substrate had different microbial communities at different periods of wheat ecosystem development and showed that the relative highest diversity of microbial communities appeared at late and middle periods of the plant ecosystem, respectively. The results also indicated that the wheat leaf and substrate had different microbial community compositions, and the wheat substrate had higher richness of microbial community than the leaf. Flavobacterium, Pseudomonas, Paenibacillus, Enterobacter, Penicillium, Rhodotorula, Acremonium, and Alternaria were dominant in the wheat leaf samples, and Pedobacter, Flavobacterium, Halomonas, Marinobacter, Salinimicrobium, Lysobacter, Pseudomonas, Halobacillus, Xanthomonas, Acremonium, Monographella, and Penicillium were dominant populations in the wheat near-root substrate samples.

Inter individual variations of the fish skin microbiota: host genetics basis of mutualism?

Directory of Open Access Journals (Sweden)

Sébastien Boutin

Full Text Available The commensal microbiota of fish skin is suspected to provide a protection against opportunist infections. The skin of fish harbors a complex and diverse microbiota that closely interacts with the surrounding water microbial communities. Up to now there is no clear evidence as to whether the host regulates the recruitment of environmental bacteria to build a specific skin microbiota. To address this question, we detected Quantitative Trait Loci (QTL associated with the abundance of specific skin microbiota bacterial strains in brook charr (Salvelinus fontinalis, combining 16S RNA tagged-amplicon 454 pyrosequencing with genetic linkage analysis. Skin microbiota analysis revealed high inter-individual variation among 86 F2 fish progeny based upon the relative abundance of bacterial operational taxonomic units (OTUs. Out of those OTUs, the pathogenic strain Flavobacterium psychrophilum and the non-pathogenic strain Methylobacterium rhodesianum explained the majority of inter-individual distances. Furthermore, a strong negative correlation was found between Flavobacterium and Methylobacterium, suggesting a mutually competitive relationship. Finally, after considering a total of 266 markers, genetic linkage analysis highlighted three major QTL associated with the abundance of Lysobacter, Rheinheimera and Methylobacterium. All these three genera are known for their beneficial antibacterial activity. Overall, our results provide evidence that host genotype may regulate the abundance of specific genera among their surface microbiota. They also indicate that Lysobacter, Rheinheimera and Methylobacterium are potentially important genera in providing protection against pathogens.

Microbiological studies on petroleum and natural gas. I. Determination of hydrocarbon-utilizing bacteria

Energy Technology Data Exchange (ETDEWEB)

Iizuka, H; Komagata, K

1964-01-01

Hydrocarbon-utilizing bacteria were isolated from oil-brine, soils etc. sampled in oil fields in Japan during 1956, and the following species were identified: Corynebacterium hydrocarboclastus nov. sp., 11 strains; Pseudomonas nitroreducens nov. sp., 1 strain; Pseudomonas maltophila Hugh and Ryschenkow, 5 strains: Brevibacterium lipolyticum (Huss) Breed, 2 strains; Pseudomonas desmolytica Gray and Thornton, 5 strains; Flavobacterium ferrugineum Sickles and Shaw, 1 strain; and Alcaligenes faecalis Chastellani and Chalmers, 1 strain. One difference between Gram-negative bacteria and Gram-positive bacteria was described on the basis of the ability of assimilating hydrocarbons.

Type IX secretion system PorM and gliding machinery GldM form arches spanning the periplasmic space

OpenAIRE

Leone, Philippe; Roche, Jennifer; Vincent, Maxence S.; Tran, Quang Hieu; Desmyter, Aline; Cascales, Eric; Kellenberger, Christine; Cambillau, Christian; Roussel, Alain

2018-01-01

Type IX secretion system (T9SS), exclusively present in the Bacteroidetes phylum, has been studied mainly in Flavobacterium johnsoniae and Porphyromonas gingivalis. Among the 18 genes, essential for T9SS function, a group of four, porK-N (P. gingivalis) or gldK-N (F. johnsoniae) belongs to a co-transcribed operon that expresses the T9SS core membrane complex. The central component of this complex, PorM (or GldM), is anchored in the inner membrane by a trans-membrane helix and interacts throug...

Flavobacteria, a Never Ending Threat for Fish: a Review

DEFF Research Database (Denmark)

Wahli, Thomas; Madsen, Lone

2018-01-01

for resistance to antibiotics. Further, newly developed vaccines and a number of new treatment methods yielded promising results, but fully convincing and generally accepted prophylactic or therapeutic methods are not yet available. Summary In summary, despite intense research in the two species and considerable......Purpose of Review In this review, we summarized the most recent findings on the partial and full genome and the phylogenetic structure of genomovars, as well as on virulence factors, vaccine development, and treatment methods of the two fish pathogenic bacteria Flavobacterium psychrophilum and F...

Immune response in rainbow trout against infection with Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Kania, Per; Madsen, Lone

farmers control the disease with antibiotics. The pathogen has a limited ability to cause disease in an experimental setting without applying a stressor. A bath-model using 150mg/L H2O2 for 60 minutes as a stressor was used on 1.4g rainbow trout fry in four experimental groups: 1) no H2O2/no bath...... infection, 2) H2O2/no bath infection, 3) no H2O2/ bath infection and 4) H2O2/ bath infection. Bath infections were carried out in 107 CFU/ml F. psychrophilum bath solution and control groups were bathed in sterile medium. Samples from all internal organs, head and skin were taken before pathogen exposure...

Genomic Diversity and Evolution of the Fish Pathogen Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Duchaud, Eric; Rochat, Tatiana; Habib, Christophe

2018-01-01

genome accounting for similar to 80% of the genes in each genome. The pan-genome seems nevertheless "open" according to the scaling exponent of a power-law fitted on the rate of new gene discovery when genomes are added one-by-one. Recombination is a key component of the evolutionary process...... of recombination and mutations to nucleotide-level differentiation (r/m) was estimated to similar to 13. Within CC-ST10, evolutionary distances computed on non-recombined regions and comparisons between 22 isolates sampled up to 27 years apart suggest a most recent common ancestor in the second half...

Sepsis neonatal por Chryseobacterium meningosepticum

Directory of Open Access Journals (Sweden)

Carlos Velásquez

2008-10-01

Full Text Available El Chrysobacterium meningosepticum conocido también como Flavobacterium meningosepticum, es una bacteria Gram negativa distribuida en la naturaleza, pero cuya variedad patógena es raramente diagnosticada. Su importancia radica en su alta resistencia antibiótica, que cuando es causa de infecciones en prematuros y adultos inmunocomprometidos conlleva a una alta mortalidad. Se realiza una revisión acerca de esta bacteria, la importancia de su diagnóstico y la vigilancia de las infecciones intrahospitalarias, a raíz de un caso presentado en una unidad de cuidados intensivos neonatal de Lima, Perú.

Ribotyping for differentiating Flavobacterium meningosepticum isolates from clinical and environmental sources

DEFF Research Database (Denmark)

Colding, H; Bangsborg, J; Fiehn, N E

1994-01-01

RI), a discriminatory index of 0.95 to 0.97 was found. The value of ribotyping in an epidemiological setting was assessed for three clinical isolates of F. meningosepticum from an outbreak of meningitis and bacteremia in the neonatal intensive care unit, Rigshospitalet, Copenhagen, Denmark. The three clinical isolates...

Flavobacterium plurextorum sp. nov. Isolated from Farmed Rainbow Trout (Oncorhynchus mykiss)

OpenAIRE

Zamora, Leydis; Fern?ndez-Garayz?bal, Jos? F.; S?nchez-Porro, Cristina; Palacios, Mari Angel; Moore, Edward R. B.; Dom?nguez, Lucas; Ventosa, Antonio; Vela, Ana I.

2013-01-01

Five strains (1126-1H-08(T), 51B-09, 986-08, 1084B-08 and 424-08) were isolated from diseased rainbow trout. Cells were Gram-negative rods, 0.7 µm wide and 3 µm long, non-endospore-forming, catalase and oxidase positive. Colonies were circular, yellow-pigmented, smooth and entire on TGE agar after 72 hours incubation at 25°C. They grew in a temperature range between 15°C to 30°C, but they did not grow at 37°Cor 42°C. Based on 16S rRNA gene sequence analysis, the isolates belonged to the genus...

Flavobacterium plurextorum sp. nov. isolated from farmed rainbow trout (Oncorhyncus mykiss)

OpenAIRE

Zamora, Leydis; Fernández Garayzábal, José F.; Sánchez-Porro Álvarez, Cristina; Palacios, Maria Ángel; Moore, Edward R. B.; Domínguez, Lucas; Ventosa Ucero, Antonio; Vela, Ana I.

2013-01-01

Five strains (1126-1H-08T, 51B-09, 986-08, 1084B-08 and 424-08) were isolated from diseased rainbow trout. Cells were Gram-negative rods, 0.7 mm wide and 3 mm long, non-endospore-forming, catalase and oxidase positive. Colonies were circular, yellow-pigmented, smooth and entire on TGE agar after 72 hours incubation at 25uC. They grew in a temperature range between 15uC to 30uC, but they did not grow at 37uCor 42uC. Based on 16S rRNA gene sequence analysis, the isolates belonged to the genus F...

Effect of heavy metals on marine Bacillus sp. and Flavobacterium sp.

Digital Repository Service at National Institute of Oceanography (India)

Nair, S.; LokaBharathi, P.A.; Chandramohan, D.

stream_size 10 stream_content_type text/plain stream_name Ecotoxicology_2_220.pdf.txt stream_source_info Ecotoxicology_2_220.pdf.txt Content-Encoding ISO-8859-1 Content-Type text/plain; charset=ISO-8859-1 ...

Characterisation of surface blebbing and membrane vesicles produced by Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Møller, Jeannette Dan; Barnes, A.C.; Dalsgaard, Inger

2005-01-01

anti-whole-cell antisera. Two distinct bands of approximately 62 and 58 kDA were highly expressed in the MVs and not seen in the OMP. MVs contained proteolytic activity towards gelatine but not towards casein and elastin, which were only degraded by live cells. Low molecular weight lipopolysaccharides...

Dietary copper effects survival of channel catfish challenged with Flavobacterium columnare

Science.gov (United States)

Columnaris disease is one of the most important bacterial diseases of channel catfish, Ictalurus punctatus, commercially grown in the US. Copper sulfate (CuSO4) has also been shown to be both therapeutic and prophylactic as a water treatment for columnaris disease. As copper is an essential dietar...

Coupling of dimethylsulfide oxidation to biomass production by a marine flavobacterium

Digital Repository Service at National Institute of Oceanography (India)

Green, D.H.; Shenoy, D.M.; Hart, M.C.; Hatton, A.D.

. Stirling, and J. R. Quayle. 1982. Co-metabolism [and discussion]. Phil Trans R Soc Lond B 297:481-496. 7. del Valle, D. A., D. J. Kieber, and R. P. Kiene. 2007. Depth-dependent fate of biologically- consumed dimethylsulfide in the Sargasso Sea. Mar Chem.... Klenk. 1985. Measurement of protein using bicinchoninic acid. Anal Biochem 150:76-85. 27. Vila-Costa, M., D. A. del Valle, J. M. Gonzalez, D. Slezak, R. P. Kiene, O. Sanchez, and R. Simo. 2006. Phylogenetic identification and metabolism of marine...

A Polyprotein-Expressing Salmonid Alphavirus Replicon Induces Modest Protection in Atlantic Salmon (Salmo Salar Against Infectious Pancreatic Necrosis

Directory of Open Access Journals (Sweden)

Azila Abdullah

2015-01-01

Full Text Available Vaccination is an important strategy for the control and prevention of infectious pancreatic necrosis (IPN in farmed Atlantic salmon (Salmo salar in the post-smolt stage in sea-water. In this study, a heterologous gene expression system, based on a replicon construct of salmonid alphavirus (SAV, was used for in vitro and in vivo expression of IPN virus proteins. The large open reading frame of segment A, encoding the polyprotein NH2-pVP2-VP4-VP3-COOH, as well as pVP2, were cloned and expressed by the SAV replicon in Chinook salmon embryo cells (CHSE-214 and epithelioma papulosum cyprini (EPC cells. The replicon constructs pSAV/polyprotein (pSAV/PP and pSAV/pVP2 were used to immunize Atlantic salmon (Salmo salar by a single intramuscular injection and tested in a subsequent IPN virus (IPNV challenge trial. A low to moderate protection against IPN was observed in fish immunized with the replicon vaccine that encoded the pSAV/PP, while the pSAV/pVP2 construct was not found to induce protection.

Detection and genetic analysis of aquabirnaviruses in subclinically infected aquarium fish.

Science.gov (United States)

Shin, Sang Phil; Gomez, Dennis Kaw; Kim, Ji Hyung; Choresca, Casiano Hermorpia; Han, Jee Eun; Jun, Jin Woo; Park, Se Chang

2011-03-01

Aquabirnaviruses (ABVs) cause serious diseases in a variety of fish species used worldwide in aquaculture and have been isolated from a variety of healthy fish and shellfish species. The type species of ABV is Infectious pancreatic necrosis virus (IPNV), which is the causative agent of a highly contagious disease in juvenile salmonid fish. Marine birnaviruses (MABVs) have been isolated from various marine fish and shellfish. In Korea, ABV infection has been identified in several fish and shellfish. The current study presents sequence data from nested polymerase chain reaction products of 3 ABV strains obtained from different species of asymptomatic aquarium fish collected from a private commercial aquarium in Korea. Phylogenetic analysis of these strains, based on the partial nucleotide sequence of the VP2/NS junction, placed them within the genogroup VII (95-99% bootstrap confidence), which also contains MABV. The subclinically infected fish may be a source of MABV infection for other susceptible fish species inside the aquarium and potentially represent a serious challenge for the management of MABV infections. Additionally, the presence of MABV in these subclinically infected aquarium fish imported from other countries indicates that there is a need for the establishment of appropriate quarantine practices.

Microbial Diversity and Parasitic Load in Tropical Fish of Different Environmental Conditions.

Directory of Open Access Journals (Sweden)

Philipp Hennersdorf

Full Text Available In this study we analysed fecal bacterial communities and parasites of three important Indonesian fish species, Epinephelus fuscoguttatus, Epinephelus sexfasciatus and Atule mate. We then compared the biodiversity of bacterial communities and parasites of these three fish species collected in highly polluted Jakarta Bay with those collected in less polluted Indonesian areas of Cilacap (E. sexfasciatus, A. mate and Thousand Islands (E. fuscoguttatus. In addition, E. fuscoguttatus from net cages in an open water mariculture facility was compared with free living E. fuscoguttatus from its surroundings. Both core and shared microbiomes were investigated. Our results reveal that, while the core microbiomes of all three fish species were composed of fairly the same classes of bacteria, the proportions of these bacterial classes strongly varied. The microbial composition of phylogenetically distant fish species, i.e. A. mate and E. sexfasciatus from Jakarta Bay and Cilacap were more closely related than the microbial composition of more phylogentically closer species, i.e. E. fuscoguttatus, E. sexfasciatus from Jakarta Bay, Cilacap and Thousand Islands. In addition, we detected a weak negative correlation between the load of selected bacterial pathogens, i.e. Vibrio sp. and Photobacterium sp. and the number of endoparasites. In the case of Flavobacterium sp. the opposite was observed, i.e. a weak positive correlation. Of the three recorded pathogenic bacterial genera, Vibrio sp. was commonly found in E. fuscoguttatus from mariculture, and lessly in the vicinity of the net cages and rarely in the fishes from the heavily polluted waters from Jakarta Bay. Flavobacterium sp. showed higher counts in mariculture fish and Photobacteria sp. was the most prominent in fish inside and close to the net cages.

Characterization of bacterial diversity in pulque, a traditional Mexican alcoholic fermented beverage, as determined by 16S rDNA analysis.

Science.gov (United States)

Escalante, Adelfo; Rodríguez, María Elena; Martínez, Alfredo; López-Munguía, Agustín; Bolívar, Francisco; Gosset, Guillermo

2004-06-15

The bacterial diversity in pulque, a traditional Mexican alcoholic fermented beverage, was studied in 16S rDNA clone libraries from three pulque samples. Sequenced clones identified as Lactobacillus acidophilus, Lactobacillus strain ASF360, L. kefir, L. acetotolerans, L. hilgardii, L. plantarum, Leuconostoc pseudomesenteroides, Microbacterium arborescens, Flavobacterium johnsoniae, Acetobacter pomorium, Gluconobacter oxydans, and Hafnia alvei, were detected for the first time in pulque. Identity of 16S rDNA sequenced clones showed that bacterial diversity present among pulque samples is dominated by Lactobacillus species (80.97%). Seventy-eight clones exhibited less than 95% of relatedness to NCBI database sequences, which may indicate the presence of new species in pulque samples.

Effect of gamma-irradiation for shelf life extension of chicken meat

International Nuclear Information System (INIS)

Prachasitthisak, Y.; Ito, Hitoshi.

1996-01-01

On the study of microbiological quality of 12 samples of chicken meat produced in several different area in Japan, total aerobic bacteria were determined as 8x10 4 to 5x10 7 per g. Coliforms were 8x10 1 to 3x10 4 per g with Escherichia, Proteus and Klebsiella. Dominant putrefactive bacteria were determined as lactic acid bacteria, Pseudomonas and Flavobacterium. The shelf life of irradiated chicken meat at 1 kGy extended more than 6 days at 10degC storage. Irradiation of chicken meat at 3 kGy extended 12 days. Coliforms were disappeared at 1 kGy irradiation. (author)

Comparative analysis of sequences from PT 2013

DEFF Research Database (Denmark)

Mikkelsen, Susie Sommer

Sheatfish and not EHNV. Generally, mistakes occurred at the ends of the sequences. This can be due to several factors. One is that the sequence has not been trimmed of the sequence primer sites. Another is the lack of quality control of the chromatogram. Finally, sequencing in just one direction can result...... diseases in Europe. As part of the EURL proficiency test for fish diseases it is required to sequence any RANA virus isolates found in any of the samples. It is also highly recommended to sequence the ISA virus to determine whether it be HPRΔ or HPR0. Furthermore, it is recommended that any VHSV and IHNV...... isolates be genotyped. As part of the evaluation of the proficiency results it was decided this year to look into the quality and similarity of the sequence results for selected viruses. Ampoule III in the proficiency test 2013 contained an EHNV isolate. The EURL received 43 sequences from 41 laboratories...

The Structural Diversity of Carbohydrate Antigens of Selected Gram-Negative Marine Bacteria

Directory of Open Access Journals (Sweden)

Elena P. Ivanova

2011-10-01

Full Text Available Marine microorganisms have evolved for millions of years to survive in the environments characterized by one or more extreme physical or chemical parameters, e.g., high pressure, low temperature or high salinity. Marine bacteria have the ability to produce a range of biologically active molecules, such as antibiotics, toxins and antitoxins, antitumor and antimicrobial agents, and as a result, they have been a topic of research interest for many years. Among these biologically active molecules, the carbohydrate antigens, lipopolysaccharides (LPSs, O-antigens found in cell walls of Gram-negative marine bacteria, show great potential as candidates in the development of drugs to prevent septic shock due to their low virulence. The structural diversity of LPSs is thought to be a reflection of the ability for these bacteria to adapt to an array of habitats, protecting the cell from being compromised by exposure to harsh environmental stress factors. Over the last few years, the variety of structures of core oligosaccharides and O-specific polysaccharides from LPSs of marine microrganisms has been discovered. In this review, we discuss the most recently encountered structures that have been identified from bacteria belonging to the genera Aeromonas, Alteromonas, Idiomarina, Microbulbifer, Pseudoalteromonas, Plesiomonas and Shewanella of the Gammaproteobacteria phylum; Sulfitobacter and Loktanella of the Alphaproteobactera phylum and to the genera Arenibacter, Cellulophaga, Chryseobacterium, Flavobacterium, Flexibacter of the Cytophaga-Flavobacterium-Bacteroides phylum. Particular attention is paid to the particular chemical features of the LPSs, such as the monosaccharide type, non-sugar substituents and phosphate groups, together with some of the typifying traits of LPSs obtained from marine bacteria. A possible correlation is then made between such features and the environmental adaptations undertaken by marine bacteria.

Wpływ szczepów bakterii wyizolowanych z hydroponicznej uprawy sałaty (Lactuca sativa L. na wzrost siewek sałaty, rosnących w obecnosci rożnych form pożywienia azotowego [Influence of bacterial strains isolated from hydroponic cultures of lettuce (Lactuca sativa L. on the growth of lettuce seedlings growing in the presence of various forms of nitrogen nutrition

Directory of Open Access Journals (Sweden)

Z. Kobierzyńska-Gołąb

2015-06-01

Full Text Available 320 bacterial strains isolated from the surface of cultivated plants, as well as from other parts of hydroponic cultures showed stimulating (49 bacterial strains or inhibitory (9 bacterial strains properties in respect to the investigated plant. The following bacteria were isolated: Pseudomonas, Flavobacterium, Agrobacterium, Achromobacter and Chromobacterium. The effects of active bacterial strains on the growth of seedlings were investigated in dependence on the kind of inorganic form of nitrogen present in the nutrient solutions. The same bacterial strains exerted a stimulating effect on seedlings growing on nitrates, weaker stimulation was observed in cultures with ammonium nitrate; the growth of lettuce seedlings on nutrient solution with ammonium only, was, as a rule, inhibited by the bacteria.

Dicty_cDB: Contig-U04013-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available pid:none) Flavobacterium psychrophilum JIP... 98 3e-19 EU678894_1( EU678894 |pid:none) Bacillus intermedius ...46 3e-04 2 ( EH672780 ) CCIL10412.b1_G11.ab1 CCI(LMS) chicory Cichorium i... 46 3e-04 2 ( EJ368820 ) 1092963715450 Global-Ocean-Sampl...41 |pid:none) Sulfolobus solfataricus P2, com... 89 2e-16 AH1770( AH1770 ) conserved hypothetical protein lin2710 [imported...AY653733 ) Acanthamoeba polyphaga mimivirus, complete genome. 62 3e-05 1 ( EH678404 ) CCIL3410.b1_D14.ab1 CC...ngs S... 44 6.8 1 >( AC116963 ) Dictyostelium discoideum chromosome 2 map 4657875-4914984 strain AX4, compl

Plant Natural compounds with antibacterial activity towards common pathogens of pond-cultured channel catfish (Ictalurus punctatus).

Science.gov (United States)

Schrader, Kevin K

2010-07-01

The bacteria Edwardsiella ictaluri and Flavobacterium columnare cause enteric septicemia and columnaris disease, respectively, in channel catfish (Ictalurus punctatus). Natural therapeutants may provide an alternative to current management approaches used by producers. In this study, a rapid bioassay identified plant compounds as potential therapeutants. Chelerythrine chloride and ellagic acid were the most toxic toward E. ictaluri, with 24-h IC50 of 7.3 mg/L and 15.1 mg/L, respectively, and MIC of 2.1 mg/L and 6.5 mg/L, respectively. Chelerythrine chloride, ellagic acid, β-glycyrrhetinic acid, sorgoleone, and wogonin were the most toxic towards two genomovars of F. columnare, and wogonin had the strongest antibacterial activity (MIC = 0.3 mg/L).

Selection of media for antimicrobial susceptibility testing of fish pathogenic bacteria

DEFF Research Database (Denmark)

Dalsgaard, Inger

2001-01-01

The available data concerning antimicrobial susceptibility testing of fish pathogens showed that there is no consensus to the basal medium currently being employed. Different media recommended for susceptibility testing of human pathogens (Mueller-Hinton Agar, Tryptone Soya Agar, Antibiotic Medium...... 3, Diagnostic Sensitivity Test Agar) have been used in addition to media (Brain Heart Infusion Agar, Heart Infusion Agar, Columbia Blood Agar) normally utilized for cultivating fastidious bacteria. When testing marine pathogens, sodium chloride or seawater has been included in the media. Media...... normally used for cultivation of pathogens with specific growth requirements like Flavobacterium species and Renibacterium salmoninarum have been used for susceptibility testing. The Mueller-Hinton Agar and different modifications of this medium was used most frequently in published studies on resistant...

Reproducible methods for experimental infection with Flavobacterium psychrophilum in rainbow trout Oncorhynchus mykiss

DEFF Research Database (Denmark)

Madsen, Lone; Dalsgaard, Inger

1999-01-01

, and this method was tested using isolates with different elastin- degrading profiles and representing different serotypes. Injecting trout, average weight 1 g, with 10(4) CFU (colony- forming units) per fish caused cumulative mortalities around 60 to 70%. The virulent strains belonged to certain serotypes...... and degraded elastin. The intraperitoneal injection challenge method could be used on larger fish, but the infection dose was 10(7) CFU per fish before mortalities occurred. Bath infection and bath infection in combination with formalin treatment (stress) seemed to be reproducible methods that could be used...

Detection of Flavobacterium psychrophilum from fish tissue and water samples by PCR amplification

DEFF Research Database (Denmark)

Wiklund, T.; Madsen, Lone; Bruun, Morten Sichlau

2000-01-01

investigation, the possible detection of Fl. psychrophilum from fish tissue and water samples was examined using nested PCR with DNA probes against a sequence of the 16S rRNA genes. The DNA was extracted using Chelex(R) 100 chelating resin. The primers, which were tested against strains isolated from diseased...... fish, healthy fish, fish farm environments and reference strains, proved to be specific for Fl. psychrophilum. The obtained detection limit of Fl. psychrophilum seeded into rainbow trout brain tissue was 0.4 cfu in the PCR tube, corresponding to 17 cfu mg(-1) brain tissue. The PCR-assay proved...... to be more sensitive than agar cultivation of tissue samples from the brain of rainbow trout injected with Fl. psychrophilum. In non-sterile fresh water seeded with Fl. psychrophilum the detection limit of the PCR- assay was 1.7 cfu in the PCR tube, corresponding to 110 cfu ml(-1) water. The PCR...

Pathophysiology of infectious hematopoietic necrosis virus disease in rainbow trout: hematological and blood chemical changes in moribund fish

Science.gov (United States)

Amend, D.F.; Smith, L.

1975-01-01

Infectious hematopoietic necrosis (IHN) is a rhabdoviral disease of rainbow trout (Salmo gairdneri). Trout were injected with IHNV, and various hematological and biochemical measurements of clinically ill fish were compared to uninfected controls. Infected fish had reduced corpuscular counts, hemoglobin, and packed cell volume, but normal mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration. The percentage of immature erythrocytes was increased, but the percentage of leukocytes was unchanged. Differential leukocyte counts showed a significant decrease in neutrophils, increase in lymphocytes, but no change in monocytes. Unidentifiable necrobiotic cells were prevelant in blood smears and hematopoietic tissue imprints. Plasma bicarbonate, chloride, calcium, phosphorus, bilirubin, and osmolality were significantly reduced, but plasma glucose and anterior kidney ascorbate were unchanged. Plasma pH increased and the alpha fractions of the serum proteins were altered. No change was found in plasma enzymes, except that a LDH isozyme was significantly increased. The alkali reserve was diminished and alterations in acid-base and fluid balance occurred. Death probably resulted from a severe electrolyte and fluid imbalance caused by renal failure.

The occurrence and prevention of Infectious Pancreatic Necrosis Virus (IPNV) in rainbow trout fish farms: the importance of improved sanitary practices

OpenAIRE

AGIM REXHEPI; KURTESH SHERIFI; PETER SCHEINERT; LINDA GRAPCI-KOTORRI

2014-01-01

The viral diseases of rainbow trout are of worldwide concern including infectious pancreatic necrosis (IPN). On this study good sanitary practices are applied to prevent outbreaks and minimize losses caused from IPN disease. To highlight importance of improvements on sanitary practices in IPN prevention were compared results from two studies, first conducted in year 2006-2009 with results from one year later study conducted in 2010. In year 2010, samples of organs are collected from 260 fish ...

Hotspots for selected metal elements and microbes accumulation and the corresponding water quality deterioration potential in an unchlorinated drinking water distribution system.

Science.gov (United States)

Liu, Gang; Tao, Yu; Zhang, Ya; Lut, Maarten; Knibbe, Willem-Jan; van der Wielen, Paul; Liu, Wentso; Medema, Gertjan; van der Meer, Walter

2017-11-01

Biofilm formation, loose deposit accumulation and water quality deterioration in drinking water distribution systems have been widely reported. However, the accumulation and distribution of harbored elements and microbes in the different niches (loose deposits, PVC-U biofilm, and HDPE biofilm) and their corresponding potential contribution to water quality deterioration remain unknown. This precludes an in-depth understanding of water quality deterioration and the development of proactive management strategies. The present study quantitatively evaluated the distribution of elements, ATP, Aeromonas spp., and bacterial communities in distribution pipes (PVC-U, D = 110 mm, loose deposit and biofilm niches) and household connection pipes (HDPE, D = 32 mm, HDPE biofilm niches) at ten locations in an unchlorinated distribution system. The results show that loose deposits in PVC-U pipes, acting as sinks, constitute a hotspot (highest total amount per meter pipe) for elements, ATP, and target bacteria groups (e.g., Aeromonas spp., Mycobacterium spp., and Legionella spp.). When drinking water distribution system niches with harbored elements and microbes become sources in the event of disturbances, the highest quality deterioration potential (QDP) is that of HDPE biofilm; this can be attributed to its high surface-to-volume ratio. 16s rRNA analysis demonstrates that, at the genus level, the bacterial communities in the water, loose deposits, PVC-U biofilm, and HDPE biofilm were dominated, respectively, by Polaromonas spp. (2-23%), Nitrosipra spp. (1-47%), Flavobacterium spp. (1-36%), and Flavobacterium spp. (5-67%). The combined results of elemental composition and bacterial community analyses indicate that different dominant bio-chemical processes might occur within the different niches-for example, iron-arsenic oxidizing in loose deposits, bio-calumniation in PVC-U biofilm, and methane oxidizing in HDPE biofilm. The release of 20% loose deposits, 20% PVC-U biofilm

Novel method reveals a narrow phylogenetic distribution of bacterial dispersers in environmental communities exposed to low hydration conditions

DEFF Research Database (Denmark)

Krüger, U. S.; Bak, F.; Aamand, J.

2018-01-01

In this study, we developed a method that provides community-level surface dispersal profiles under controlled hydration conditions from environmental samples and enables us to isolate and uncover the diversity of the fastest bacterial dispersers. The method expands on the Porous Surface Model (PSM...... Pseudomonas putida and Flavobacterium johnsoniae strains from their non-motile mutants. Applying the method to soil and lake water bacterial communities showed that community-scale dispersal declined as conditions became drier. However, for both communities, dispersal was detected even under low hydration...... dispersers were substantially less diverse than the total communities. The dispersing fraction of the soil microbial community was dominated by Pseudomonas which increased in abundance at low hydration conditions, while the dispersing fraction of the lake community was dominated by Aeromonas and, under wet...

Quorum sensing signal molecules (acylated homoserine lactones) in Gram-negative fish pathogenic bacteria

DEFF Research Database (Denmark)

Bruhn, Jesper Bartholin; Dalsgaard, Inger; Nielsen, K.F.

2005-01-01

The aim of the present study was to investigate the production of quorum sensing signals (specifically acylated homoserine lactones, AHLs) among a selection of strains of Gram-negative fish bacterial pathogens. These signals are involved in the regulation of virulence factors in some human...... salmonicida and Vibrio splendidus were also positive. Aeromonas species produced N-butanoyl homoserine lactone (BHL) and N-hexanoyl homoserine lactone (HHL) and 1 additional product, whereas N-3-oxo-hexanoyl homoserine lactone (OHHL) and HHL were detected in Vibrio salmonicida. N-3-oxo-octanoyl homoserine...... lactone (OOHL) and N-3-octanoyl homoserine lactone (OHL) were detected in Y. ruckeii. AHLs were not detected from strains of Photobacterium damselae, Flavobacterium psychrophilum or Moritella viscosa. AHLs were extracted from fish infected with Y. ruckeri but not from fish infected with A. salmonicida...

Gamma radiation effects on liofilized human serum

International Nuclear Information System (INIS)

Padron Soler, E.; Romay Penabad, Z.; Chavez Ardanza, A.; Hernandez Gonzalez, C.; Martin Gonzalez, O.; Garcia Gonzalez, I.; Prieto Miranda, E.

1995-01-01

Human freeze dried serum was artificially contaminated with Flavobacterium sp. for studying the effects of gamma radiation of it. The radiobiological parameters of the contaminator were determined and the sterilization dose was set. The quality of the product irradiated at both, calculated sterilization dose (8.5 kGy) an another one about 25 kGy was determined. It was made according to: sterility testing, total proteins, pH enzymes (alanina-aminotransferase, aspartato-aminotransferase, alkaline phosphatase), protein electrophoresis, fast performance liquid chromatographic and effect on the cellular growth. From the latter was concluded that the calculated sterilization dose was adequate form keeping the biological properties and viability of the irradiated serum. Nevertheless, the dose of 25 k Gy was not adequate because of its dangerous effects on the cell culture

Effect of Hydrogen Peroxide on Immersion Challenge of Rainbow Trout Fry with Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Madsen, Lone; Dalsgaard, Inger

2013-01-01

and produce high levels of mortality attempts to establish a reproducible immersion model have been less successful. Various concentrations of hydrogen peroxide (H2O2) were evaluated before being used as a pre-treatment stressor prior to immersion exposure to F. psychrophilum. H2O2 accelerated the onset...... of mortality and increased mortality approximately two-fold; from 9.1% to 19.2% and from 14.7% to 30.3% in two separate experiments. Clinical signs observed in the infected fish corresponded to symptoms characteristically seen during natural outbreaks. These findings indicate that pre-treatment with H2O2 can...

Hematology and productive performance of nile tilapia (Oreochromis niloticus naturally infected with Flavobacterium columnare

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F. A Sebastião

2011-03-01

Full Text Available Columnaris disease is one of the main causes of mortality in tilapia rearing and is responsible for large economic losses worldwide. Hematology is a tool that makes it possible to study organisms' physiological responses to pathogens. It may assist in making diagnoses and prognoses on diseases in fish populations. The hematological variables of nile tilapia were studied in specimens with a clinical diagnosis of columnaris disease and in specimens that were disease-free. The total erythrocyte count, hemoglobin rate, hematocrit percentage, mean corpuscular hemoglobin (MCH, mean corpuscular hemoglobin concentration (MCHC, mean corpuscular volume (MCV, organic defense blood cell percentages (leukocytes and thrombocytes and hepatosomatic and splenosomatic index were determined. The results showed that there were changes in the erythrocytic series and in organic defense blood cells, in the fish infected with the bacterium, with reductions in erythrocytic variables and significant increases in the numbers of circulating lymphocytes and neutrophils.

Isolation, Screening and Development of Local Bacterial Consortia With Azo Dyes Decolourising Capability

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Khadijah, O.

2009-01-01

Full Text Available A total of 1540 bacterial isolates were isolated and screened for their ability to degrade selected azo dyes. Of these, nine isolates were chosen for further studies based on their ability to degrade a wide spectrum of dyes efficiently and rapidly. Several microbial consortia were developed and tested for their effectiveness. Overall the consortia were able to degrade 70 - 100% colour within 72 hours compared to 60 – 97% colour removed by individual isolates. A microbial consortium labelled C15 showed good growth in agitation culture but the colour removal was best in static culture with 80 - 100% colour removed in less than 72 hours. Based on the 16S rRNA sequencing, two of the bacterial isolates in C15 belong to the Chryseobacterium genus while the other one belongs to Flavobacterium genus.

Empedobacter brevis Meningitis in a Neonate: A Very Rare Case of Neonatal Meningitis and Literature Review

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Deepak Sharma

2016-01-01

Full Text Available Empedobacter brevis is gram-negative bacilli that belongs to Flavobacteriaceae family. It was previously known with name of Flavobacterium breve. The reservoir of these bacteria is soil, plants, water, food, hospital water sources, including incubators, sinks, faucets, tap water, hemodialysis systems, saline solutions, and other pharmaceutical solutions. We report a case of term female newborn, admitted with complaint of respiratory distress developing soon after birth and developed clinical features of sepsis at age of 92 hours of postnatal life. The sepsis screen was positive and blood culture and cerebrospinal fluid showed growth of Empedobacter brevis that was resistant to multiple antibiotics. The neonate was treated with appropriate antibiotics and was discharged successfully. The novelty of the case report is that this is the first case report of neonatal sepsis caused by Empedobacter brevis.

Molecular analysis of bacterial populations in water samples from two Uranium mill tailings by using RISA retrieval

International Nuclear Information System (INIS)

Selenska-Pobell, S.; Radeva, G.

2004-01-01

Ribosomal intergenic spacer amplification (RISA) retrieval was applied to analyse the natural bacterial communities in drain waters of two uranium mill tailings - Gittersee/Coschuetz in Germany and Shiprock in the USA. About 35% of the clones from RISA library constructed for the samples of the German tailings represented a microdiverse population of Planctomycetales. The rest of the clones were affiliated with rather diverse bacterial groups including γ- and δ-Proteobacteria, Cytophaga/Flavobacterium/Bacteroides (CFB), Nitrospira, Verrucomicrobia and Actinobacteria. 8% of the cloned sequences represented a novel bacterial lineage from the recently described division NC3. Bacterial diversity in the Shiprock mill tailings was found to be significantly lower. RISA library constructed for those samples contained only two larger groups of clones representing β-proteobacterial species and one small group which was affiliated with δ-Proteobacteria. (authors)

Molecular analysis of bacterial populations in water samples from two Uranium mill tailings by using RISA retrieval

Energy Technology Data Exchange (ETDEWEB)

Selenska-Pobell, S [Forschungszentrum Rossendorf, Institute of Radiochemistry, Dresden (Germany); Radeva, G [Bulgarian Academy of Sciences, Institute of Molecular Biology, Sofia (Bulgaria)

2004-07-01

Ribosomal intergenic spacer amplification (RISA) retrieval was applied to analyse the natural bacterial communities in drain waters of two uranium mill tailings - Gittersee/Coschuetz in Germany and Shiprock in the USA. About 35% of the clones from RISA library constructed for the samples of the German tailings represented a microdiverse population of Planctomycetales. The rest of the clones were affiliated with rather diverse bacterial groups including {gamma}- and {delta}-Proteobacteria, Cytophaga/Flavobacterium/Bacteroides (CFB), Nitrospira, Verrucomicrobia and Actinobacteria. 8% of the cloned sequences represented a novel bacterial lineage from the recently described division NC3. Bacterial diversity in the Shiprock mill tailings was found to be significantly lower. RISA library constructed for those samples contained only two larger groups of clones representing {beta}-proteobacterial species and one small group which was affiliated with {delta}-Proteobacteria. (authors)

Bacterial communities in the rhizosphere of amilaceous maize (Zea mays L. as assessed by pyrosequencing

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David Correa-Galeote

2016-07-01

Full Text Available Maize (Zea mays L. is the staple diet of the native peasants in the Quechua region of the Peruvian Andes who continue growing it in small plots called chacras following ancestral traditions. The abundance and structure of bacterial communities associated with the roots of amilaceous maize has not been studied in Andean chacras. Accordingly, the main objective of this study was to describe the rhizospheric bacterial diversity of amilaceous maize grown either in the presence or the absence of bur clover cultivated in soils from the Quechua maize belt. Three 16S rRNA gene libraries, one corresponding to sequences of bacteria from bulk soil of a chacra maintained under fallow conditions, the second from the rhizosphere of maize-cultivated soils, and the third prepared from rhizospheric soil of maize cultivated in intercropping with bur clover were examined using pyrosequencing tags spanning the V4 and V5 hypervariable regions of the gene. A total of 26031 sequences were found that grouped into 5955 distinct operational taxonomic units which distributed in 309 genera. The numbers of OTUs in the libraries from the maize-cultivated soils were significantly higher than those found in the libraries from bulk soil. One hundred ninety seven genera were found in the bulk soil library and 234 and 203 were in those from the maize and maize/bur clover-cultivated soils. Sixteen out of the 309 genera had a relative abundance higher than 0.5% and the were (in decreasing order of abundance Gp4, Gp6, Flavobacterium, Subdivision3 genera incertae sedis of the Verrucomicrobia phylum, Gemmatimonas, Dechloromonas, Ohtaekwangia, Rhodoferax, Gaiella, Opitutus, Gp7, Spartobacteria genera incertae sedis, Terrimonas, Gp5, Steroidobacter and Parcubacteria genera incertae sedis. Genera Gp4 and Gp6 of the Acidobacteria, Gemmatimonas and Rhodoferax were the most abundant in bulk soil, whereas Flavobacterium, Dechloromonas and Ohtaekwangia were the main genera in the rhizosphere

A distinct bacterial dysbiosis associated skin inflammation in ovine footrot

Science.gov (United States)

Maboni, Grazieli; Blanchard, Adam; Frosth, Sara; Stewart, Ceri; Emes, Richard; Tötemeyer, Sabine

2017-03-01

Ovine footrot is a highly prevalent bacterial disease caused by Dichelobacter nodosus and characterised by the separation of the hoof horn from the underlying skin. The role of innate immune molecules and other bacterial communities in the development of footrot lesions remains unclear. This study shows a significant association between the high expression of IL1β and high D. nodosus load in footrot samples. Investigation of the microbial population identified distinct bacterial populations in the different disease stages and also depending on the level of inflammation. Treponema (34%), Mycoplasma (29%) and Porphyromonas (15%) were the most abundant genera associated with high levels of inflammation in footrot. In contrast, Acinetobacter (25%), Corynebacteria (17%) and Flavobacterium (17%) were the most abundant genera associated with high levels of inflammation in healthy feet. This demonstrates for the first time there is a distinct microbial community associated with footrot and high cytokine expression.

Isolation and Identification of Phosphate Solubilizing and Nitrogen Fixing Bacteria from Soil in Wamena Biological Garden, Jayawijaya, Papua

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SRI WIDAWATI

2005-07-01

Full Text Available A study was undertaken to investigate the occurrence of phosphate solubilizing bacteria (PSB and nitrogen-fixing bacteria (NFB from soil samples of Wamena Biological Garden (WbiG. Eleven soil samples were collected randomly to estimate microbial population which used plate count method. The result showed that the microbial population ranged from 5.0x103-7.5x106 cells of bacteria/gram of soil and 5.0x103-1.5x107 cells of bacteria/gram of soil for PSB and NFB respectively. There were 17 isolates which have been identified till genus and species. The isolated microorganism were identified as PSB i.e. Bacillus sp., B. pantothenticus, B. megatherium, Flavobacterium sp., F. breve, Klebsiella sp., K. aerogenes, Chromobacterium lividum, Enterobacter alvei, E. agglomerans, Pseudomonas sp., Proteus sp. and as NFB i.e. Azotobacter sp., A. chroococcum, A. paspalii, Rhizobium sp., and Azospirillum sp.

Effect of gamma rays on gram negative bacterial endotoxin

International Nuclear Information System (INIS)

Hilmy, N.; Dzulkarnain, B.

1977-01-01

Escherichia coli br and a Flavobacterium sp., isolated from Tjiptomangunkusumo Hospital in Jakarta, were used as bacterial endotoxin (pyrogen) source. Suspensoion of 10 7 , 10 6 , and 10 5 bacteria in sterile and pyrogen free Sodium Chloride Injection, that had been treated by heat at 80 deg C for 15 minutes or treated by irradiation with a dose of 2.5 x 10 6 rad, were injected each to nine rabbits. Maximum temperature rises were registered and compared. Data were interpreted according to the requirements of Indonesian Pharmacopea (1972) and British Pharmacopea (1973). The results showed that a dose of 2.5 x 10 6 rad did not reduce the pyrogenic activity. The rate of bacterial multiplications in the pharmaceutical preparations were tested by using sterile Dextrose (2.5%) and Sodium Chloride Injection as medium; storage time were 0; 6; and 24 hours. (author)

A selective and differential medium for Vibrio harveyi.

Science.gov (United States)

Harris, L; Owens, L; Smith, S

1996-01-01

A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated from the larval rearing water of a commercial prawn hatchery with V. harveyi agar as a primary isolation medium and were positively identified, by conventional tests, as V. harveyi. This agar displays great potential as a primary isolation medium and offers significant advantages over thiosulfate-citrate-bile salts-sucrose agar as a medium for differentiating V. harveyi from other marine and estuarine Vibrio species. PMID:8795252

Microbiological activities in a shallow-ground repository with cementitious wasteform

International Nuclear Information System (INIS)

Varlakova, G.A.; Dyakonova, A.T.; Netrusov, A.I.; Ojovan, M.I.

2012-01-01

Cementitious wasteform with immobilised nuclear power plant operational radioactive waste disposed in a near surface testing repository for about 20 years have been analysed for microbiological activities. Clean cultures were selected from the main metabolic groups expected within repository environment e.g. anaerobic de-nitrifying, fermenting, sulphur-reducing, iron-reducing, and oxidizing, thio-bacterium and mushrooms. Microbiological species were identified within cementitious wasteform, in the clayey soil near the wasteform and in the contacting water. The most populated medium was the soil with microbial populations Bacillus, Pseudomonas and Micrococcus, and densities of populations up to 3.6*10 5 colony/g. Microbial populations of generic type Bacillus, Pseudomonas, Rhodococcus, Alcaligenes, Micrococcus, Mycobacterium, and Arthrobacter were identified within cementitious wasteform. Populations of Arthrobacter, Pseudomonas, Alcaligenes, Rhodococcus, Bacillus and Flavobacterium were identified in the water samples contacting the cementitious wasteform. Microbiological species identified are potential destructors of cementitious wasteform and containers. (authors)

Epidemiological cut-off values for Flavobacterium psychrophilum MIC data generated by a standard test protocol

DEFF Research Database (Denmark)

Smith, P.; Endris, R.; Kronvall, G.

2016-01-01

-1 for florfenicol, ≤0.025 mg L-1 for oxolinic acid (OXO), ≤0.125 mg L-1 for oxytetracycline and ≤20 (1/19) mg L-1 for trimethoprim/sulphamethoxazole. For ampicillin and amoxicillin, the majority of putative wild-type observations were 'off scale', and therefore, statistically valid cut-off values...

BIORREMEDIACIÓN DE UN SUELO CON DIESEL MEDIANTE EL USO DE MICROORGANISMOS AUTÓCTONOS

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ARRIETA RAMÍREZ OLGA MARIA

2012-07-01

Full Text Available En este estudio, se aisló y caracterizó bioquímica y molecularmente un consorcio bacteriano capaz de degradar los diferentes hidrocarburos presentes en un combustible diesel,conformado por los siguientes géneros: Enterobacter sp, Bacillus sp, Staphylococcus aureus, Sanguibacter soli, Arthrobacter sp y Flavobacterium sp, a partir de un suelo contaminado con diesel a escala de laboratorio, y tratado mediante 2 tecnologías de biorremediación: atenuación natural y bioestimulación. Se definió como parámetro de control la concentración de Hidrocarburos Totales del Petróleo (HTP y para el cual, se obtuvo una reducción en la concentración en un periodo de 4 meses de 36,86% para atenuación natural y 50,99% para bioestimulación. La medición de la eficiencia de remoción de hidrocarburos se cuantificó por cromatografía de gases acoplada a masas (GC-MS.

Biological nutrient removal and molecular biological characteristics in an anaerobic-multistage anaerobic/oxic (A-MAO) process to treat municipal wastewater.

Science.gov (United States)

Huang, Xiao; Dong, Wenyi; Wang, Hongjie; Jiang, Shilong

2017-10-01

This study aimed to present an anaerobic-multistage anaerobic/oxic (A-MAO) process to treat municipal wastewater. The average COD, NH 4 + -N, TN, and TP removal efficiency were 91.81%, 96.26%, 83.73% and 94.49%, respectively. Temperature plunge and C/N decrease have a certain impact on the modified process. Characteristics of microbial community, function microorganism, and correlation of microbial community with environmental variables in five compartments were carried out by Illumina Miseq high-throughput sequencing. The differences of microbial community were observed and Blastocatella, Flavobacterium and Pseudomonas were the dominant genus. Nitrosomonas and Nitrospira occupied a dominant position in AOB and NOB, respectively. Rhodospirillaceae and Rhodocyclaceae owned a considerable proportion in phosphorus removal bacteria. DO and COD played significant roles on affecting the microbial components. The A-MAO process in this study demonstrated a high potential for nutrient removal from municipal wastewater. Copyright © 2017 Elsevier Ltd. All rights reserved.

Dynamic Changes in Bacterial Population and Corresponding Exoenzyme Activity in Response to a Tropical Phytoplankton Bloom Chattonella marina

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Anit M. Thomas

2014-01-01

Full Text Available The raphidophyte Chattonella marina (Subrahmanyan Hara & Chihara bloom which causes lethal effects on marine ecosystem has been reported intermittently from Indian waters. In the present study, periodic samplings were made in a Chattonella marina bloom area, off Mahe, on 27 and 29 October and 1 November 2011 (in different phases of the bloom to assess the associated bacterial population and their exoenzyme activity. Microbial community composition of Chattonella marina bloom revealed a twentyfold increase in bacterial load over the nonbloom area. The bacterial genera, Micrococcus, Flavobacterium, Vibrio, and Pseudomonas, increased significantly during the declining phase of the bloom. An assessment of the extracellular enzyme production also showed a marked increase in percentage of bacterial strains, potent in protease production, suggesting the possible role of proteolytic bacteria in bloom crash. This study reveals the bacterial community succession during the bloom and indicates that bacteria play an important role in bloom regulation.

The Screw-Like Movement of a Gliding Bacterium Is Powered by Spiral Motion of Cell-Surface Adhesins.

Science.gov (United States)

Shrivastava, Abhishek; Roland, Thibault; Berg, Howard C

2016-09-06

Flavobacterium johnsoniae, a rod-shaped bacterium, glides over surfaces at speeds of ∼2 μm/s. The propulsion of a cell-surface adhesin, SprB, is known to enable gliding. We used cephalexin to generate elongated cells with irregular shapes and followed their displacement in three dimensions. These cells rolled about their long axes as they moved forward, following a right-handed trajectory. We coated gold nanoparticles with an SprB antibody and tracked them in three dimensions in an evanescent field where the nanoparticles appeared brighter when they were closer to the glass. The nanoparticles followed a right-handed spiral trajectory on the surface of the cell. Thus, if SprB were to adhere to the glass rather than to a nanoparticle, the cell would move forward along a right-handed trajectory, as observed, but in a direction opposite to that of the nanoparticle. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Characterization of the Bacterial Community Naturally Present on Commercially Grown Basil Leaves: Evaluation of Sample Preparation Prior to Culture-Independent Techniques

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Siele Ceuppens

2015-08-01

Full Text Available Fresh herbs such as basil constitute an important food commodity worldwide. Basil provides considerable culinary and health benefits, but has also been implicated in foodborne illnesses. The naturally occurring bacterial community on basil leaves is currently unknown, so the epiphytic bacterial community was investigated using the culture-independent techniques denaturing gradient gel electrophoresis (DGGE and next-generation sequencing (NGS. Sample preparation had a major influence on the results from DGGE and NGS: Novosphingobium was the dominant genus for three different basil batches obtained by maceration of basil leaves, while washing of the leaves yielded lower numbers but more variable dominant bacterial genera including Klebsiella, Pantoea, Flavobacterium, Sphingobacterium and Pseudomonas. During storage of basil, bacterial growth and shifts in the bacterial community were observed with DGGE and NGS. Spoilage was not associated with specific bacterial groups and presumably caused by physiological tissue deterioration and visual defects, rather than by bacterial growth.

Shifts in microbial community structure and diversity in a MBR combined with worm reactors treating synthetic wastewater.

Science.gov (United States)

Liu, Jia; Zuo, Wei; Zhang, Jun; Li, Hui; Li, Lipin; Tian, Yu

2017-04-01

The chemical oxygen demand (COD) and NH 3 -N removal, membrane fouling, sludge characteristics and microbial community structure in a membrane bioreactor (MBR) coupled with worm reactors (SSBWR) were evaluated for 210days. The obtained results were compared to those from a conventional MBR (C-MBR) operated in parallel. The results indicated that the combined MBR (S-MBR) achieved higher COD and NH 3 -N removal efficiency, slower increase in membrane fouling, better sludge settleability and higher activities of the related enzymes in the activated sludge. Denaturing gradient gel electrophoresis was used to analyze the microbial community structures in the C-MBR and the S-MBR. The microbial community structure in the S-MBR was more diverse than that in the C-MBR. Additionally, the slow-growing microbes such as Saprospiraceae, Actinomyces, Frankia, Clostridium, Comamonas, Pseudomonas, Dechloromonas and Flavobacterium were enriched in the S-MBR, further accounting for the sludge reduction, membrane fouling alleviation and wastewater treatment. Copyright © 2016. Published by Elsevier B.V.

Diversity of both the cultivable protease-producing bacteria and bacterial extracellular proteases in the coastal sediments of King George Island, Antarctica.

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Ming-Yang Zhou

Full Text Available Protease-producing bacteria play a vital role in degrading sedimentary organic nitrogen. However, the diversity of these bacteria and their extracellular proteases in most regions remain unknown. In this paper, the diversity of the cultivable protease-producing bacteria and of bacterial extracellular proteases in the sediments of Maxwell Bay, King George Island, Antarctica was investigated. The cultivable protease-producing bacteria reached 10(5 cells/g in all 8 sediment samples. The cultivated protease-producing bacteria were mainly affiliated with the phyla Actinobacteria, Firmicutes, Bacteroidetes, and Proteobacteria, and the predominant genera were Bacillus (22.9%, Flavobacterium (21.0% and Lacinutrix (16.2%. Among these strains, Pseudoalteromonas and Flavobacteria showed relatively high protease production. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria were serine proteases or metalloproteases. These results begin to address the diversity of protease-producing bacteria and bacterial extracellular proteases in the sediments of the Antarctic Sea.

Flavobacterium nitratireducens sp. nov., an amylolytic bacterium of the family Flavobacteriaceae isolated from coastal surface seawater

Digital Repository Service at National Institute of Oceanography (India)

Nupur; Bhumika, V.; Srinivas, T.N.R.; AnilKumar, P.

A novel Gram-negative, rod-shaped, non-motile bacterium, designated strain N1 sup(T), was isolated from a marine water sample collected from the sea shore, Bay of Bengal, Visakhapatnam, India. The strain was positive for starch hydrolysis, nitrate...

Phage-Host Interactions in Flavobacterium psychrophilum and the Potential for Phage Therapy in Aquaculture

DEFF Research Database (Denmark)

Christiansen, Rói Hammershaimb

, the increasing problem with antibiotic resistance has led to increased attention to the use of phages for controlling F. psychrophilum infections in aquaculture. In a synopsis and four scientific papers, this PhD project studies the potential and optimizes the use of phage therapy for treatment and prevention......, studies of the genetic diversity and susceptibility patterns of F. psychrophilum strains and phages isolated in three geographically distinct areas (Chile, Denmark, and USA) showed that the strains and phages clustered into geographically distinct groups. However, cross-infectivity between Chilean phage......-phage. In the third paper, a detailed analysis of the resistance mechanisms in F. psychrophilum and six phage resistant mutants was done. The results revealed unique changes in the genomes in all the phage resistant strains and that some of these changes were related to cell surface properties which were suggested...

Influence of organic diets and probiotics on an experimental Flavobacterium psychrophilum infection in rainbow trout fry

DEFF Research Database (Denmark)

Madsen, Lone; Ingerslev, Hans-Christian; Boye, Mette

2012-01-01

following exposure to pathogenic microorganisms. The aim of the Danish project OPTIFISH is to optimize growth and survival for organic cultured rainbow trout. OPTIFISH investigates how organic vs. non-organic diet types as well as diets with or without probiotics affect the intestine, the intestinal...... of fry to pathogens, e.g. F .psychrophilum. In the current experiment four diet types were tested on fry, a conventional type (Inicio®, BioMar A/S) with and without probiotics, as well as an organic type with and without probiotic. Bactocell® (Lallemand) was used as the probiotic. Studies were done...... been fed diets that were organic or conventional nor if the feed had been added probiotic. No significance in mortalities was seen between the diet groups. The bacteriological examinations showed that the bacterium had entered the fish, as it was found in inner organs in one fourth of the sampled fish...

Immune Response in Head Kidney of Rainbow Trout Fry Following Stress and Infection with Flavobacterium Psychrophilum

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Kania, P. W.; Madsen, Lone

response in rainbow trout against F. psychrophilum in order to create preventive measures against RTFS. A limited number of studies have been carried out so far and have relied on samples from either naturally infected or injection-challenged fish. The use of naturally infected fish introduces many...... before pathogen exposure to elevate mortality. The model was used to examine the immune response to infection in rainbow trout fry (≈1 g); both with and without preceding H2O2 treatment. Samples from the head kidney were taken before pathogen exposure and 4 hours, 48 hours, 125 hours and 192 hours after...... exposure. The regulation of several immune relevant genes was examined and the relative bacterial load was assessed. Although it is not determined how H2O2 increases mortality, it is assumed to be due to stress. Exposure to H2O2 prior to infection altered the regulation of several genes, and several...

Chemical resistance of the gram-negative bacteria to different sanitizers in a water purification system

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Penna Thereza CV

2006-08-01

Full Text Available Abstract Background Purified water for pharmaceutical purposes must be free of microbial contamination and pyrogens. Even with the additional sanitary and disinfecting treatments applied to the system (sequential operational stages, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were isolated and identified from a thirteen-stage purification system. To evaluate the efficacy of the chemical agents used in the disinfecting process along with those used to adjust chemical characteristics of the system, over the identified bacteria, the kinetic parameter of killing time (D-value necessary to inactivate 90% of the initial bioburden (decimal reduction time was experimentally determined. Methods Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were called in house (wild bacteria. Pseudomonas diminuta ATCC 11568, Pseudomonas alcaligenes INCQS , Pseudomonas aeruginosa ATCC 15442, Pseudomonas fluorescens ATCC 3178, Pseudomonas picketti ATCC 5031, Bacillus subtilis ATCC 937 and Escherichia coli ATCC 25922 were used as 'standard' bacteria to evaluate resistance at 25°C against either 0.5% citric acid, 0.5% hydrochloric acid, 70% ethanol, 0.5% sodium bisulfite, 0.4% sodium hydroxide, 0.5% sodium hypochlorite, or a mixture of 2.2% hydrogen peroxide (H2O2 and 0.45% peracetic acid. Results The efficacy of the sanitizers varied with concentration and contact time to reduce decimal logarithmic (log10 population (n cycles. To kill 90% of the initial population (or one log10 cycle, the necessary time (D-value was for P. aeruginosa into: (i 0.5% citric acid, D = 3.8 min; (ii 0.5% hydrochloric acid, D = 6.9 min; (iii 70% ethanol, D = 9.7 min; (iv 0.5% sodium bisulfite, D = 5.3 min; (v 0.4% sodium hydroxide, D = 14.2 min; (vi 0.5% sodium

Bactérias gram negativas resistentes a antimicrobianos em alimentos Gram-negative bacteria resistant to antibiotics in foods

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José Cavalcante de Albuquerque Ribeiro Dias

1985-12-01

Full Text Available A partir de 154 espécimens de alimentos, representados por hortaliças (alface, leite e merenda escolar, obteve-se o isolamento e identificação de 400 amostras de bacilos Gram negativos. Esta amostragem se distribuiu em 339 enterobactérias (Escherichia, Shigella, Citrobacter, Klebsiella, Enterobacter, Serratia e Proteus e 61 de gêneros afins (Acinetobacter, Flavobacterium, Aeromonas e Pseudomonas. Submetendo-se as culturas aos antimicrobianos: sulfadiazina (Su, estreptomicina (Sm, tetraciclina (Tc, cloranfenicol (Cm, canamicina (Km, ampicilina (Ap, ácido nalidíxico (Nal e gentamicina (Gm, observou-se apenas seis estirpes sensíveis a todas as drogas e sensibilidade absoluta à Gm. A predominância dos modelos Su (27,6% e Su-Ap (39,6% incidiu nas enterobactérias, enquanto que, 18,0% para Ap e 9,8% para Su-Ap foram detectados nos gêneros afins. Para caracterização da resistência foram realizados testes de conjugação e a totalidade das culturas não revelou transferência para o gene que confere resistência ao ácido nalidíxico. Relevantes são as taxas de amostras R+ observadas nos bacilos entéricos, oscilando em torno de 90% (leite e merenda escolar e alface, em torno de 70%From 154 food samples, including vegetables (lettuce, milk and meals served at school it was possible to isolate and identify 400 Gram negative bacilli distributed among 339 enteric bacteria (Escherichia, Shigella, Citrobacter, Klebsiella, Enterobacter, Serratia and Proteus and other 61 non enteric bacilli (Acinetobacter, Flavobacterium, Aeromonas and Pseudomonas. Submitting this cultures to the drugs sulfadiazine (Su, streptomycin (Sm, tetracycline (Tc, chloramphenicol (Cm, kanamycin (Km, ampicillin (Ap, nalidixic acid (Nal and gentamycin (Gm it was observed only six stocks susceptible to all drugs and total sensibility to Gm. Among enteric bacteria the profiles Su (27,6% and Su-Ap (39,6% predominated, while for the non enteric bacilli percentages of 18.0 for

Inhibitory effect of aromatic geranyl derivatives isolated from Heliotropium filifolium on infectious pancreatic necrosis virus replication.

Science.gov (United States)

Modak, Brenda; Sandino, Ana María; Arata, Loredana; Cárdenas-Jirón, Gloria; Torres, René

2010-02-24

Infectious pancreatic necrosis is a disease caused by a birnavirus affecting several wild and commercial aquatic organisms. This infectious disease results in significant losses in the farming industry and therefore effective therapeutic agents are needed to control outbreaks caused by this pathogen. Our goal was to evaluate in vitro antiviral effect of a group of natural compounds (geranyl aromatic derivatives) isolated from the resinous exudate of the plant Heliotropium filifolium (Heliotropiaceae), semi-synthetics compounds obtained from them, and the resinous exudate, on CHSE-214 cell line infected with infectious pancreatic necrosis virus (IPNV) using a virus plaque inhibition assay at various concentrations. The compound ester filifolinyl senecionate was the best antiviral with EC(50) 160 microg/mL and a cytotoxic concentration required to reduce cell viability by 50% up to 400 microg/mL. In order to obtain information about the mechanism of the antiviral action, was evaluated the influence of ester filifolinyl senecionate on the viral RNA synthesis. This compound produced inhibition of the synthesis of viral genomic RNA, suggesting that the ester could be interacting with the viral RNA during the viral cycle. Additionally, a preliminary study of the interaction between ester and a sample of single-stranded RNA was studied at the level of theory Restricted Hartree Fock PM3 method. The results showed that the ester formed hydrogen bonds mainly with nitrogenous bases but not with ribose and phosphate. These results allow propose that the ester filifolinyl senecionate is a good candidate for used as antiviral therapy for IPN virus in salmon fry. Copyright 2009 Elsevier B.V. All rights reserved.

Quality control ranges for testing broth microdilution susceptibility of Flavobacterium columnare and F. psychrophilum to nine antimicrobials

DEFF Research Database (Denmark)

Gieseker, Charles M.; Mayer, Tamara D.; Crosby, Tina C.

2012-01-01

salmonicida subsp. salmonicida ATCC 33658 against 10 antimicrobials (ampicillin, enrofloxacin, erythromycin, florfenicol, flumequine, gentamicin, ormetoprim/sulfadimethoxine, oxolinic acid, oxytetracycline, and trimethoprim/sulfamethoxazole) in diluted (4 g l−1) cation-adjusted Mueller-Hinton broth incubated...

Comparative study of wastewater treatment and nutrient recycle via activated sludge, microalgae and combination systems.

Science.gov (United States)

Wang, Liang; Liu, Jinli; Zhao, Quanyu; Wei, Wei; Sun, Yuhan

2016-07-01

Algal-bacterial synergistic cultivation could be an optional wastewater treatment technology in temperate areas. In this study, a locally screened vigorous Chlorella strain was characterized and then it was used in a comparative study of wastewater treatment and nutrient recycle assessment via activated sludge (AS), microalgae and their combination systems. Chlorella sp. cultured with AS in light showed the best performance, in which case the removal efficiencies of COD, NH3-N and TP were 87.3%, 99.2% and 83.9%, respectively, within a short period of 1day. Algal-bacterial combination in light had the best settleability. Chlorella sp. contained biomass, could be processed to feed, fertilizer or fuel due to the improved quality (higher C/H/N) compared with sludge. PCR-DGGE analysis shows that two types of rhizobacteria, namely, Pseudomonas putida and Flavobacterium hauense were enriched in sludge when cultured with algae in light, serving as the basics for artificial consortium construction for improved wastewater treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Assessment of bacterial diversity in Hyalomma aegyptium, H. marginatum and H. excavatum ticks through tag-encoded pyrosequencing.

Science.gov (United States)

Keskin, Adem; Bursali, Ahmet; Snow, David E; Dowd, Scot E; Tekin, Saban

2017-12-01

Ticks are among the most significant human-biting ectoparasites and they play a major role in transmission of many pathogenic agents to humans. In the present study, three species of Hyalomma ticks, Hyalomma aegyptium, H. marginatum and H. excavatum, were examined for the presence of zoonotic bacteria, both male and female ticks alike. Examination of microbial diversity with tag-encoded pyrosequencing indicates that H. marginatum and H. excavatum were more diversity rich than H. aegyptium. Although numerous pathogenic and non-pathogenic bacterial genera were detected, including Acidovorax, Bacillus, Bacteroides, Bdellovibrio, Clostridium, Curvibacter, Escherichia, Flavobacterium, Limnohabitans, Paenibacillus, Ralstonia, Sarcina, Sediminibacterium, Segetibacter Stenotrophomonas and Variovorax, the predominant zoonotic bacteria represented in these ticks were genera Borrelia, Francisella, and Rickettsia. To the authors' knowledge, this work represents the first detection of Yersinia enterocolitica in the tick H. excavatum, raising questions regarding the vector competency of this tick, as well as associations of different disease representations perhaps through previously unforeseen routes of pathogen introduction. Likewise, similar questions are related to the presence of Legionella pneumophila in one H. excavatum sample.

A Meta-Analysis of the Bacterial and Archaeal Diversity Observed in Wetland Soils

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Xiaofei Lv

2014-01-01

Full Text Available This study examined the bacterial and archaeal diversity from a worldwide range of wetlands soils and sediments using a meta-analysis approach. All available 16S rRNA gene sequences recovered from wetlands in public databases were retrieved. In November 2012, a total of 12677 bacterial and 1747 archaeal sequences were collected in GenBank. All the bacterial sequences were assigned into 6383 operational taxonomic units (OTUs 0.03, representing 31 known bacterial phyla, predominant with Proteobacteria (2791 OTUs, Bacteroidetes (868 OTUs, Acidobacteria (731 OTUs, Firmicutes (540 OTUs, and Actinobacteria (418 OTUs. The genus Flavobacterium (11.6% of bacterial sequences was the dominate bacteria in wetlands, followed by Gp1, Nitrosospira, and Nitrosomonas. Archaeal sequences were assigned to 521 OTUs from phyla Euryarchaeota and Crenarchaeota. The dominating archaeal genera were Fervidicoccus and Methanosaeta. Rarefaction analysis indicated that approximately 40% of bacterial and 83% of archaeal diversity in wetland soils and sediments have been presented. Our results should be significant for well-understanding the microbial diversity involved in worldwide wetlands.

Type IX secretion system PorM and gliding machinery GldM form arches spanning the periplasmic space.

Science.gov (United States)

Leone, Philippe; Roche, Jennifer; Vincent, Maxence S; Tran, Quang Hieu; Desmyter, Aline; Cascales, Eric; Kellenberger, Christine; Cambillau, Christian; Roussel, Alain

2018-01-30

Type IX secretion system (T9SS), exclusively present in the Bacteroidetes phylum, has been studied mainly in Flavobacterium johnsoniae and Porphyromonas gingivalis. Among the 18 genes, essential for T9SS function, a group of four, porK-N (P. gingivalis) or gldK-N (F. johnsoniae) belongs to a co-transcribed operon that expresses the T9SS core membrane complex. The central component of this complex, PorM (or GldM), is anchored in the inner membrane by a trans-membrane helix and interacts through the outer membrane PorK-N complex. There is a complete lack of available atomic structures for any component of T9SS, including the PorKLMN complex. Here we report the crystal structure of the GldM and PorM periplasmic domains. Dimeric GldM and PorM, each contain four domains of ~180-Å length that span most of the periplasmic space. These and previously reported results allow us to propose a model of the T9SS core membrane complex as well as its functional behavior.

Effect of radurization on microbial flora of vacuum-packaged trout (Salmo gairdneri)

Energy Technology Data Exchange (ETDEWEB)

Ehlermann, D; Diehl, J F; Hussain, A M [Bundesforschungsanstalt fuer Ernaehrung, Karlsruhe (Germany, F.R.). Inst. fuer Strahlentechnologie

1976-11-01

The microbial spoilage pattern was studied for nonirradiated and irradiated (100 and 200 krad) vacuum-packed trout during storage under melting crushed ice. Total microbial count from original nonirradiated fish before storage amounted to 5 x 10/sup 3//g. The value increased to 5 x 10/sup 8//g for the vacuum-packed nonirradiated sample after one month storage. At the end of the same period, total counts in fish treated with 100 and 200 krad increased to 10/sup 7/ and 5 x 10/sup 6/ per g, respectively. Microorganisms isolated from nonirradiated starting material, in order of decreasing number, consisted of Pseudomonas, Flavobacterium, Achromobacter-Moraxella, yeasts, Bacillus, Proteus, Aeromonas, Micrococcus, and few Lactobacillus and Corynebacterium. During storage and with the program of spoilage, the pattern of the microbial flora was shifted. Pseudomonas, initially dominating, were almost eliminated after two weeks storage, while Bacillus and Proteus became predominant. In the irradiated fish, regardless of the dose, Achromobacter-Moraxella, Micrococcus and Lactobacillus became dominant throughout the storage period.

Changes in the microflora of Vienna sausages after irradiation with gamma-rays and storage at 10/sup 0/C

Energy Technology Data Exchange (ETDEWEB)

Ito, H; Sato, T [Japan Atomic Energy Research Inst., Takasaki, Gunma. Takasaki Radiation Chemistry Research Establishment

1973-02-01

The species of microorganisms which can grow on commercial viennas on storage at 10/sup 0/C were Lactobacillus, Streptococcus, and yeasts. When the viennas specially made which did not contain preservatives in it were used for this investigation, growth of microorganisms such as Lactobacillus, Streptococcus, Micrococcus, Bacillus, and yeasts were predominant on storage at 10/sup 0/C, and Pseudomonas and molds some time propagated. When smoked-viennas specially made for the National Project were used for preservation, growth of microorganisms consisted mainly of the species of Lactobacillus, Micrococcus, Acinetobacter, Flavobacterium, Streptococcus, Serratia, Corynebacterium, and yeasts. Irradiation of viennas at 300 and 500 krad reduced the aforementioned flora to the Lactobacillus, Streptococcus, Acinetobacter, and yeasts. The number of microorganisms on the viennas packed with nitrogen gas was not increased for 3 to 7 days by means of 300 krad irradiation, and extended the storage-life 2 to 3 times. When irradiated with a dose of 500 krad, the number of microorganisms was not increased for 9 to 14 days on storage at 10/sup 0/C.

Microorganism selection and performance in bioslurry reactors treating PAH-contaminated soil.

Science.gov (United States)

Cassidy, D P; Hudak, A J

2002-09-01

A continuous-flow reactor (CSTR) and a soil slurry-sequencing batch reactor (SS-SBR) were operated in 81 vessels for 200 days to treat a soil contaminated with polycyclic aromatic hydrocarbons (PAH). Filtered slurry samples were used to quantify bulk biosurfactant concentrations and PAH emulsification. Concentrations of Corynebacterium aquaticum, Flavobacterium mizutaii, Mycobacterium gastri, Pseudomonas aeruginosa, and Pseudomonas putida were determined using fatty acid methyl ester (FAME) analysis. The CSTR and SS-SBR selected microbial consortia with markedly different surfactant-producing and PAH-degrading abilities. Biosurfactant levels in the SS-SBR reached 4 times the critical micelle concentration (CMC) that resulted in considerable emulsification of PAH. In contrast, CSTR operation resulted in nomeasurable biosurfactant production. Total PAH removal efficiency was 93% in the SS-SBR, compared with only 66% in the CSTR, and stripping of PAH was 3 times less in the SS-SBR. Reversing the mode of operation on day 100 caused a complete reversal in microbial consortia and in reactor performance by day 140. These results show that bioslurry reactor operation can be manipulated to control overall reactor performance.

Structure of nitrogen-converting communities induced by hydraulic retention time and COD/N ratio in constantly aerated granular sludge reactors treating digester supernatant.

Science.gov (United States)

Cydzik-Kwiatkowska, Agnieszka; Rusanowska, Paulina; Zielińska, Magdalena; Bernat, Katarzyna; Wojnowska-Baryła, Irena

2014-02-01

This study investigated how hydraulic retention time (HRT) and COD/N ratio affect nitrogen-converting consortia in constantly aerated granules treating high-ammonium digester supernatant. Three HRTs (10, 13, 19 h) were tested at COD/N ratios of 4.5 and 2.3. Denaturing gradient gel electrophoresis and relative real-time PCR were used to characterize the microbial communities. When changes in HRT and COD/N increased nitrogen loading, the ratio of the relative abundance of aerobic to anaerobic ammonium-oxidizers decreased. The COD/N ratio determined the species composition of the denitrifiers; however, Thiobacillus denitrificans, Pseudomonas denitrificans and Azoarcus sp. showed a high tolerance to the environmental conditions and occurred in the granules from all reactors. Denitrifier genera that support granule formation were identified, such as Pseudomonas, Shinella, and Flavobacterium. In aerated granules, nirK-possessing bacteria were more diverse than nirS-possessing bacteria. At a low COD/N ratio, N2O-reducer diversity increased because of the presence of bacteria known as aerobic denitrifiers. Copyright © 2013 Elsevier Ltd. All rights reserved.

A protein secretion system linked to bacteroidete gliding motility and pathogenesis

Science.gov (United States)

Sato, Keiko; Naito, Mariko; Yukitake, Hideharu; Hirakawa, Hideki; Shoji, Mikio; McBride, Mark J.; Rhodes, Ryan G.; Nakayama, Koji

2009-01-01

Porphyromonas gingivalis secretes strong proteases called gingipains that are implicated in periodontal pathogenesis. Protein secretion systems common to other Gram-negative bacteria are lacking in P. gingivalis, but several proteins, including PorT, have been linked to gingipain secretion. Comparative genome analysis and genetic experiments revealed 11 additional proteins involved in gingipain secretion. Six of these (PorK, PorL, PorM, PorN, PorW, and Sov) were similar in sequence to Flavobacterium johnsoniae gliding motility proteins, and two others (PorX and PorY) were putative two-component system regulatory proteins. Real-time RT-PCR analysis revealed that porK, porL, porM, porN, porP, porT, and sov were down-regulated in P. gingivalis porX and porY mutants. Disruption of the F. johnsoniae porT ortholog resulted in defects in motility, chitinase secretion, and translocation of a gliding motility protein, SprB adhesin, to the cell surface, providing a link between a unique protein translocation system and a motility apparatus in members of the Bacteroidetes phylum. PMID:19966289

Nutrients removal and bacterial community structure for low C/N municipal wastewater using a modified anaerobic/anoxic/oxic (mA2/O) process in North China.

Science.gov (United States)

Zhang, Shihua; Huang, Zhijia; Lu, Shujian; Zheng, Jun; Zhang, Xinxi

2017-11-01

A modified anaerobic/anoxic/oxic (mA2/O) process based on utilizing the internal carbon source and adding polypropylene carriers was operated for 90d to investigate the nutrients removal performance and bacterial community. This system exhibited a stable and efficient performance, particularly, in removing the NH 4 + -N and total phosphorus. The results of high-throughput sequencing showed that the 13 dominant genera containing Pseudomonas, Comamonas, Arcobacter, Nitrobacteria, Nitrosospira, Nitrosomonas, Bacteroides, Flavobacterium, Rhizobium, Acinetobacter, Zoogloea, Rhodocyclus and Moraxella were shared by five zones, inferring that they were the essential players in treating low C/N (below 5.0) municipal wastewater around 10°C. The average abundance of Nitrosospira (4.21%) was higher than that of Nitrosomonas (2.93%), suggested that Nitrosospira performed well under low temperature for nitrification. Additionally, both known Rhodocyclus-related PAOs and GAOs Competibacter were not detected possibly due to low temperature. Redundancy analysis (RDA) indicated that DO played more important roles in regulating bacterial community composition than HRT. Copyright © 2017 Elsevier Ltd. All rights reserved.

Marine Bacteria with antimicrobials capacity isolated from cultures of bivalve mollusks

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Fabiola Pellon

2014-06-01

Full Text Available Microorganisms have commonly been studied as producers of antibacterial substances; yet they are also considered producers of antifungic, antiviral, antiparasitic, citotoxics and inhibitory of other forms of cellular growth substances. This paper describes the isolation, inhibitory potential and phenotipic characterization of native bacterial strains associated to bivalve mollusks such as Argopecten purpuratus “concha de abanico” and Crassostrea gigas “ostra” in cultivation systems. From 345 marine strains collected, 20 strains were recovered that had the ability of inhibiting a wide spectrum of fish, mollusks and shellfish pathogenic bacteria; being the most sensitive pathogens Aeromonas sobria P-281, Aeromonas hydrophila ATCC 7966, Vibrio vulnificus ATCC 27562 and Vibrio parahaemolyticus ATCC 17803. The phenotipic characterization of this strains with inhibitory capacity allowed the identification of the following genera: Vibrio (40%, Aeromonas (15%, Flavobacterium (10%, Pseudomonas (5%, Moraxella (5%, Flexibacter (5%. A 20% could not be identified. The results suggest that the isolated bacteria could be used as probiotics agents for the biological control of pathogens from marine organisms of interest in mariculture.

Localization and characterization of acharan sulfate in the body of the giant African snail Achatina fulica.

Science.gov (United States)

Jeong, J; Toida, T; Muneta, Y; Kosiishi, I; Imanari, T; Linhardt, R J; Choi, H S; Wu, S J; Kim, Y S

2001-12-01

Acharan sulfate is a glycosaminoglycan (GAG), having the structure -->4)-2-acetamido-2-deoxy-alpha-D-glucopyranose(1-->4)-2-sulfo-alpha-L-idopyranosyluronic acid (1-->, isolated from the body of the giant African snail Achatina fulica. This GAG represents 3-5% of the dry weight of this snail's soft body tissues. Frozen sections and polyester wax sections of the snail's body were stained by Alcian blue-periodic acid-Schiff's reagent (PAS) to localize acharan sulfate. Alcian blue staining indicated that GAG was mainly secreted into the outer surface of the body from internal granules. A highly mucous material was collected and treated and the acharan sulfate was recovered by ethanol and cetyl pyridinium chloride precipitation. Crude acharan sulfate was purified by DEAE-Sephacel ion-exchange chromatography. Depolymerization of intact mucus and purified acharan sulfate fractions by heparin lyase II (heparitinase I) from Flavobacterium heparinum produced an unsaturated disaccharide as a major product, establishing the repeating unit of acharan sulfate. These results demonstrate that mucus in the granule and secreted to the outside of the body is composed entirely of acharan sulfate.

Development of electrochemical biosensor for detection of pathogenic microorganism in Asian dust events.

Science.gov (United States)

Yoo, Min-Sang; Shin, Minguk; Kim, Younghun; Jang, Min; Choi, Yoon-E; Park, Si Jae; Choi, Jonghoon; Lee, Jinyoung; Park, Chulhwan

2017-05-01

We developed a single-walled carbon nanotubes (SWCNTs)-based electrochemical biosensor for the detection of Bacillus subtilis, one of the microorganisms observed in Asian dust events, which causes respiratory diseases such as asthma and pneumonia. SWCNTs plays the role of a transducer in biological antigen/antibody reaction for the electrical signal while 1-pyrenebutanoic acid succinimidyl ester (1-PBSE) and ant-B. subtilis were performed as a chemical linker and an acceptor, respectively, for the adhesion of target microorganism in the developed biosensor. The detection range (10 2 -10 10  CFU/mL) and the detection limit (10 2  CFU/mL) of the developed biosensor were identified while the response time was 10 min. The amount of target B. subtilis was the highest in the specificity test of the developed biosensor, compared with the other tested microorganisms (Staphylococcus aureus, Flavobacterium psychrolimnae, and Aquabacterium commune). In addition, target B. subtilis detected by the developed biosensor was observed by scanning electron microscope (SEM) analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Distribution of bacteria in frozen shrimps and their decontamination by gamma irradiation

Energy Technology Data Exchange (ETDEWEB)

Ito, Hitoshi; Adulyatham, P.; Ishigaki, Isao

1987-11-01

Six samples of frozen shrimps were obtained through importing company mainly from South-east Asian countries. Five samples of frozen shrimps contained 1.5 x 10/sup 5/ to 6.2 x 10/sup 6/ per gram of total aerobic bacteria, and 4 samples contained 1.4 x 10/sup 2/ to 2.2 x 10/sup 3/ per gram of coliforms, which are higher contamination than Japanese standard of hygenic level for frozen food products (total counts below 10/sup 5/ per gram and no coliforms). The dominant bacteria in total counts were consisted with psychrotrophic Moraxella, Arthrobacter, Micrococcus, Acinetobacter, Flavobacterium, Corynebacterium, Pseudomonas and etc, in all kinds of frozen shrimps. Salmonellae or enteric Vibrio were could not isolated by ordinary isolation methods, and there were isolated many strains of Proteus mirabilis instead of these bacteria. Necessary dose of gamma irradiation for decontamination of bacteria to hygenic level was decided as 3 approx. 5 kGy from the inactivation curves of total bacteria, coliforms, Proteus mirabilis and Salmonella typhymurium in frozen shrimps. Off-odor was suppressed by frozen condition of irradiation.

Distribution of bacteria in frozen shrimps and their decontamination by gamma irradiation

International Nuclear Information System (INIS)

Ito, Hitoshi; Adulyatham, P.; Ishigaki, Isao

1987-01-01

Six samples of frozen shrimps were obtained through importing company mainly from South-east Asian countries. Five samples of frozen shrimps contained 1.5 x 10 5 to 6.2 x 10 6 per gram of total aerobic bacteria, and 4 samples contained 1.4 x 10 2 to 2.2 x 10 3 per gram of coliforms, which are higher contamination than Japanese standard of hygenic level for frozen food products (total counts below 10 5 per gram and no coliforms). The dominant bacteria in total counts were consisted with psychrotrophic Moraxella, Arthrobacter, Micrococcus, Acinetobacter, Flavobacterium, Corynebacterium, Pseudomonas and etc, in all kinds of frozen shrimps. Salmonellae or enteric Vibrio were could not isolated by ordinary isolation methods, and there were isolated many strains of Proteus mirabilis instead of these bacteria. Necessary dose of gamma irradiation for decontamination of bacteria to hygenic level was decided as 3 ∼ 5 kGy from the inactivation curves of total bacteria, coliforms, Proteus mirabilis and Salmonella typhymurium in frozen shrimps. Off-odor was suppressed by frozen condition of irradiation. (author)

Changes in the microflora of Vienna sausages after irradiation with gamma-rays and storage at 10 deg C

International Nuclear Information System (INIS)

Ito, Hitoshi; Sato, Tomotaro

1973-01-01

The species of microorganisms which can grow on commercial viennas on the storage at 10 deg C were Lactobacillus, Streptococcus and yeasts. When the viennas specially made which did not contain preservatives in it were used for this investigation, growth of microorganisms such as Lactobacillus, Streptococcus, Micrococcus, Bacillus and yeasts were predominant on the storage at 10 deg C, and Pseudomonas and molds some time propagated. When smoked-viennas specially made for the National Project were used for preservation, growth of microorganisms consisted mainly of the species of Lactobacillus, Micrococcus, Acinetobacter, Flavobacterium, Streptococcus, Serratia, Corynebacterium and yeasts. Irradiation of viennas at 300 and 500 krad reduced the aforementioned flora to the Lactobacillus, Streptococcus, Acinetobacter and yeasts. The number of microorganisms on the viennas packed with nitrogen gas was not increased for 3 to 7 days by means of 300 krad irradiation, and extended the storage-life 2 to 3 times. When irradiated with a dose of 500 krad, the number of microorganisms was not increased for 9 to 14 days on the storage at 10 deg C. (author)

Growth and biosurfactant synthesis by Nigerian hydrocarbon-degrading estuarine bacteria

Directory of Open Access Journals (Sweden)

Sunday A Adebusoye

2008-12-01

Full Text Available The ability of microorganisms to degrade petroleum hydrocarbons is important for finding an environmentally-friendly method to restoring contaminated environmental matrices. Screening of hydrocarbon-utilizing and biosurfactant-producing abilities of organisms from an estuarine ecosystem in Nigeria, Africa, resulted in the isolation of five microbial strains identified as Corynebacterium sp. DDv1, Flavobacterium sp. DDv2, Micrococcus roseus DDv3, Pseudomonas aeruginosa DDv4 and Saccharomyces cerevisae DDv5. These isolates grew readily on several hydrocarbons including hexadecane, dodecane, crude oil and petroleum fractions. Axenic cultures of the organisms utilized diesel oil (1.0 % v/v with generation times that ranged significantly (t-test, P La capacidad de los microorganismos para degradar hidrocarburos del petróleo es de gran importancia para hallar un método aceptable y ambientalmente amigable para la restauración de terrenos ambientalmente contaminados. Al investigar las capacidades de los organismos de un ecosistema de estuario que utilizan hidrocarburos y producen biosurfactantes, se produjo como resultado el aislamiento de cinco cepas microbianas identificadas como Corynebacterium sp. DDv1, Flavobacterium sp. DDv2, Micrococcus roseus DDv3, Pseudomonas aeruginosa y DDv4 Saccharomyces cerevisiae DDv5. Estas cepas crecieron fácilmente en varios hidrocarburos incluyendo hexadecanos, dodecanos, petróleo crudo y fracciones de petróleo. Los cultivos axénicos de organismos utilizaron diesel (1.0% v/v con períodos por generación con ámbitos significativos (t-test, P <0.05 de entre 3.25 y 3.88 días, con la consiguiente producción de bio-surfactantes. La cinética del crecimiento indica que la síntesis de bio-surfactante se produjo principalmente durante la fase de crecimiento exponencial, lo que sugiere que las moléculas bioactivas son metabolitos primarios. Las cepas DDv1 y DDv4 fueron evidentemente las más metab

Bacteriophage Resistance Mechanisms in the Fish Pathogen Flavobacterium psychrophilum: Linking Genomic Mutations to Changes in Bacterial Virulence Factors

DEFF Research Database (Denmark)

Castillo, Daniel; Christiansen, Rói Hammershaimb; Dalsgaard, Inger

2015-01-01

-resistant strains had all obtained unique insertions and/or deletions and point mutations distributed among intergenic and genic regions. Mutations in genes related to cell surface properties, gliding motility, and biosynthesis of lipopolysaccharides and cell wall were found. The observed links between phage...

Hydrogen peroxide as pre-treatment stressor in experimental immer-sion challenge of rainbow trout fry with Flavobacterium psychrophilum

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Madsen, Lone; Dalsgaard, Inger

2012-01-01

. Non-medical therapeutic substances are routinely used against pathogens in aquacultures, including copper sulphate, chloramine-T, sodium carbonates, sodium chloride, formalin and hydrogen peroxide (H2O2). One of the more successful immersion models used formalin as a stressor, but a less harmful...

The pyloric caeca area is a major site for IgM(+ and IgT(+ B cell recruitment in response to oral vaccination in rainbow trout.

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Natalia A Ballesteros

Full Text Available Although previous studies have characterized some aspects of the immune response of the teleost gut in response to diverse pathogens or stimuli, most studies have focused on the posterior segments exclusively. However, there are still many details of how teleost intestinal immunity is regulated that remain unsolved, including the location of IgM(+ and IgT(+ B cells along the digestive tract and their role during the course of a local stimulus. Thus, in the current work, we have studied the B cell response in five different segments of the rainbow trout (Oncorhynchus mykiss digestive tract in both naïve fish and fish orally vaccinated with an alginate-encapsulated DNA vaccine against infectious pancreatic necrosis virus (IPNV. IgM(+ and IgT(+ cells were identified all along the tract with the exception of the stomach in naïve fish. While IgM(+ cells were mostly located in the lamina propria (LP, IgT(+ cells were primarily localized as intraepithelial lymphocytes (IELs. Scattered IgM(+ IELs were only detected in the pyloric caeca. In response to oral vaccination, the pyloric caeca region was the area of the digestive tract in which a major recruitment of B cells was demonstrated through both real time PCR and immunohistochemistry, observing a significant increase in the number of both IgM(+ and IgT(+ IELs. Our findings demonstrate that both IgM(+ and IgT(+ respond to oral stimulation and challenge the paradigm that teleost IELs are exclusively T cells. Unexpectedly, we have also detected B cells in the fat tissue associated to the digestive tract that respond to vaccination, suggesting that these cells surrounded by adipocytes also play a role in mucosal defense.

Phenotypic and molecular characterization of bacterial populations isolated from diesel-contaminated soil and treated by two bioremediation technologies

International Nuclear Information System (INIS)

Arrieta R, Olga M; Rivera R, Angela P; Rojano, Benjamin; Ruiz, Orlando; Correa, Margarita M; Cienfuegos Gallet, Astrid V; Arias, Lida; Cardona G, Santiago A

2012-01-01

In this study bioremediation is presented as an alternative for the recovery of contaminated ecosystems. In this work an experimental diesel spill on pasture land was remediated using two bioremediation technologies: natural attenuation, which is the natural capability of indigenous microorganisms to degrade a xenobiotic component in a determined time, and biostimulation, which consist in the acceleration of the degradation process through the stimulation of the metabolism of indigenous microorganisms by the addition of nutrients (P and N) to the media. Results of respirometry assays indicated that both treatments produced significant levels of hydrocarbon removal but the biostimulation treatment stranded out with 98.17% degradation. Seven bacterial isolates were obtained from these treatments which according to their molecular characterization and phylogenetic analysis belong to the genus: Enterobacter, Bacillus, Arthrobacter, Sanguibacter, Staphylococcus and Flavobacterium. All isolates were able to metabolize diesel as a carbon and energy source; for this reason and taking into account that for some of these microorganisms their role in bioremediation have not been extensively studied, it is recommended to continue with their evaluation to know their real potential for the solution of environmental problems.

Biodiversity of culturable heterotrophic bacteria in the Southern Adriatic Sea Italian coastal waters

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Loredana Stabili

2004-04-01

Full Text Available The qualitative and quantitative composition of culturable heterotrophic bacteria in water samples from the Southern Adriatic Sea of Italy was examined. Water samples were collected monthly, for a year, at 16 stations along the coast line between Brindisi and Santa Maria di Leuca. The results obtained described the heterotrophic bacterial community over an annual cycle. Mean values of bacterial densities were 5.3 x 104 CFUml-1 in Brindisi, 5.8 x 104 CFUml-1 in S. Cataldo, 4.3 x 104 CFUml-1 in Otranto and 6.7 x 104 CFUml-1 in S. M. di Leuca. The differences in bacterial densities between the sites considered were estimated. The hydrodynamic circulation, the trophism and the geographical position of the examined sites contribute to justify the different bacterial density trends. The bacterial community consisted mainly of the genera Aeromonas, Pseudomonas, Photobacterium and Flavobacterium. The Enterobacteriaceae represented a considerable fraction of the bacterial community in the Southern Adriatic Sea. Bacilli were predominant among the Gram positive bacteria. The enzymatic versatility of the observed genera suggest their importance in organic matter turnover of this oligotrophic ecosystem.

Microbialproperty improvement of saline-alkali soil for vegetable cultivation in Shanghai coastal area and its evaluation

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KOU Yiming

2015-10-01

Full Text Available In order to improve the fertility of saline-alkali soil in Shanghai coastal area,and make it suitable for vegetable cultiration,in the study,the saline-alkali soil was mixed with organic fertilizer,and then sprayed with composite microbes,which have the ability of the synergistically degrading organic substrate.The results showed that the saline-alkali soil added with 5∶1 organic fertilizer can rapidly increase the utilization ability soil organic matter.The soil microbial populations and microbial diversity index were significantly improved when applied with the 0.5% composite microbial liquid which containeds 1∶3∶3∶1 of Bacillus licheniformis,Pseudomonas sp., Flavobacterium sp.and Sphingomonas sp..At the same time,the enzymology indicators of soil urease,phosphatase,cellulase and catalase increased significantly.The vegetable cultivation experiments showed that:the biomass of Brassica chinensis nearly doubled in the original saline-alkali soil,while the yield of organic fertilizer increased 30.2% after 50 days.The research result on of the biological improvement for saline-alkali soil will have good application value in vegetable planting in coastal saline-alkali soil.

IDENTIFICATION AND BIOLOGICAL ACTIVITY OF POTENTIAL PROBIOTIC BACTERIA ISOLATED FROM POECILIA RETICULATA (GUPPY

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Aparna Balakrishna

2014-08-01

Full Text Available Antagonistic activities against candidate indicator strains, adhesion to mucus and biofilm formation of potential probiotic strains isolated from Poecilia reticulata were evaluated. Four isolated strains (MBTU_PB1, MBTU_PB2, MBTU_PB3 and MBTU_PB4 showed moderate to strong antagonistic activities against the tested five indicator strains (Aeromonas hydrophila1739, Vibrio cholera 3906, Flavobacterium 2495, Acinetobacter 1271 and Alcaligenes 1424 and these isolates were further identified using biochemical tests and 16S rDNA gene sequence analysis. Except the whole cell product, the other three cellular components, namely, heat-killed whole cell product, intracellular product and extracellular product of all the four selected isolates were equally effective, as revealed by the zone of inhibitions to the tested indicator strains. The in vitro adhesion property or the ability of colonization is often considered as a selection criteria for probiotics. All the selected four strains had higher adhesion abilities than the indicator strains. Further, these four strains had the ability to form biofilms on polystyrene surfaces. The in vitro characterization of these four strains suggests possibility of using the isolates, as individual strain or in combination, for probiotic therapy in aquaculture.

Effect of gamma irradiation on microbiological quality of japanese chicken meat and microflora change of irradiated chicken

International Nuclear Information System (INIS)

Prachasitthisak, Y.; Ito, H.

1996-01-01

The impact of gamma irradiation with doses between 0 and 8 kGy on microbiological quality of chicken meat produced in Japan and micro flora change of irradiated chicken meat were studied. Radiation at the dose 2 kGy resulted in 4 log cycles reduction of total aerobic bacteria, 5 - 6 log cycles reduction of lactic acid bacteria and 2 log cycles reduction of fungi and yeasts. For the coliforms, it could be eliminated below detectable level by irradiation dose of 1 kGy. For the chicken flora-analysis, it was found that chicken of each area had their own specific microbial community structure. Flavobacterium and Pseudomonas were found to be dominant organisms in the microflora of Japanese chicken meat. Irradiation with dose 2 kGy resulted in disappearance of Lactobacillus and Pseudomonas. The microorganisms which dominated in irradiated chickens with doses of 2 kGy and higher were Psychrobacter and yeast. These studies support the view that radiation improves the microbiological quality of chicken meat and substantiate that radiation does not present hazard resulting from a change in the microflora of irradiated chicken

Cross sectional evaluation of the gut-microbiome metabolome axis in an Italian cohort of IBD patients.

Science.gov (United States)

Santoru, Maria Laura; Piras, Cristina; Murgia, Antonio; Palmas, Vanessa; Camboni, Tania; Liggi, Sonia; Ibba, Ivan; Lai, Maria Antonia; Orrù, Sandro; Blois, Sylvain; Loizedda, Anna Lisa; Griffin, Julian Leether; Usai, Paolo; Caboni, Pierluigi; Atzori, Luigi; Manzin, Aldo

2017-08-25

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract of uncertain origin, which includes ulcerative colitis (UC) and Crohn's disease (CD). The composition of gut microbiota may change in IBD affected individuals, but whether dysbiosis is the cause or the consequence of inflammatory processes in the intestinal tissue is still unclear. Here, the composition of the microbiota and the metabolites in stool of 183 subjects (82 UC, 50 CD, and 51 healthy controls) were determined. The metabolites content and the microbiological profiles were significantly different between IBD and healthy subjects. In the IBD group, Firmicutes, Proteobacteria, Verrucomicrobia, and Fusobacteria were significantly increased, whereas Bacteroidetes and Cyanobacteria were decreased. At genus level Escherichia, Faecalibacterium, Streptococcus, Sutterella and Veillonella were increased, whereas Bacteroides, Flavobacterium, and Oscillospira decreased. Various metabolites including biogenic amines, amino acids, lipids, were significantly increased in IBD, while others, such as two B group vitamins, were decreased in IBD compared to healthy subjects. This study underlines the potential role of an inter-omics approach in understanding the metabolic pathways involved in IBD. The combined evaluation of metabolites and fecal microbiome can be useful to discriminate between healthy subjects and patients with IBD.

UJI COBA TEKNOLOGI BIOFILM KONSORSIUM BAKTERI PADA REAKTOR SEMIANAEROB-AEROB UNTUK PENGOLAHAN AIR LIMBAH DI INDUSTRI PENCELUPAN TEKSTIL SKALA RUMAH TANGGA

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Dewa Ketut Sastrawidana

2013-04-01

Full Text Available Penelitian ini bertujuan untuk menganalisis efektifitas teknologi biofilm konsorsium bakteri pada  reaktor semianaerob-aerob ntuk mengolah air limbah pencelupan tekstil. Bakteri pada reaktor semianaerob terdiri dari  Aeromonas sp. Pseudomonas sp, dan Flavobacterium sp. sedangkan pada reaktor aerob terdiri dari Vibrio sp. Plesiomonas sp. dan Enterobacter sp. Perombakan proses pertumbuhan terlekat diawali dengan menumbuhkan konsorsium bakteri pada masing-masing reaktor selama 10  hari menggunakan pada batu vulkanik merah sebagai media pelekatan bakteri. Setelah terbentuk biofilm,selanjutnya digunakan untuk merombak limbah denagn waktu tinggal limbah 2 hari. Hasil penelitian menunjukkan teknologi biofilm cukup efektif diaplikasikan pada skala lapang menghasilkan efisiensi perombakan TSS, BOD dan COD secara berturut-turut sebesar 84,7%; 80,56% dan 90,40%. Uji toksisitas air limbah tekstil menggunakan ikan nila dengan waktu paparan 3 hari menunjukkan bahwa air limbah tekstil sebelum diolah berkatagori toksik ringan dengan nilai EC50 adalah 88,80% sedangkan setelah diolah dalam reaktor biofilm konsorsium bakteri sistem anaerob-aerob selama 2 hari menjadi katagori tidak toksik dengan nilai EC50 sebesar 101,64%. Dengan demikian, pengolahan limbah tektil dengan sistem kombinasi anaerob-aerob menghasilkan kualitas limbah dengan kriteria sudah memenuhi baku mutu untuk dibuang ke lingkungan.

The impact of engineered cobalt, iron, nickel and silver nanoparticles on soil bacterial diversity under field conditions

International Nuclear Information System (INIS)

Shah, Vishal; Collins, Daniel; Shah, Shreya; Walker, Virginia K

2014-01-01

Our understanding of how engineered nanoparticles (NPs) migrate through soil and affect microbial communities is scarce. In the current study we examined how metal NPs, including those from the iron triad (iron, cobalt and nickel), moved through pots of soil maintained under winter field conditions for 50 days, when mesophilic bacteria may not be dividing. Based on total metal analysis, cobalt and nickel were localized in the top layer of soil, even after exposure to high precipitation and freeze–thaw cycles. In contrast, a bimodal distribution of silver was observed. Due to high endogenous levels of iron, the migration pattern of these NPs could not be determined. Pyrosequence analysis of the bacterial communities revealed that there was no significant engineered NP-mediated decline in microbial richness. However, analysis of individual genera showed that Sphingomonas and Lysobacter were represented by fewer sequences in horizons containing elevated metal levels whereas there was an increase in the numbers of Flavobacterium and Niastella. Collectively, the results indicate that along with the differential migration behavior of NPs in the soil matrix, their impact on soil bacterial diversity appears to be dependent on environmental parameters. (paper)

On the reaction of some bacteria and fungi on coal tar creosote. Zur Verhalten einiger Bakterien und Pilze gegenueber Steinkohlenteeroel

Energy Technology Data Exchange (ETDEWEB)

Schmidt, O.; Dittberner, D.; Faix, O. (Universitaet Hamburg, Hamburg (Germany). Ordinariat fuer Holzbiologie)

1991-01-01

To contribute to the waste management of wood preservatives, the biodegradability of coal tar creosote by bacteria and fungi has been investigated. Microorganisms comprised 24 bacterial strains and 31 fungi from different systematic and ecological groups as well as isolates from contaminated soils. Based on countings of viable cells, the experiments with various nutrient media, methods of cultivation, preservative concentrations, and organic solvents yielded some bacteria which could grow in the presence of creosote: {ital Aeromonas hydrophila}, {ital Flavobacterium} sp., {ital Pseudomonas arvilla}, {ital P. fluorescens}, and {ital P. putida}. The white-rot fungi {ital Bjerkandera adusta}, {ital Heterobasidion annosum}, {ital Hirschioporus abietinus}, {ital Lentinula edodes}, {ital Peniophora gigantea}, {ital Pleurotus ostreatus}, {ital Schizophyllum commune}, and {ital Trametes versicolor}, the brown-rot fungus {ital Lentinus lepideus}, the staining fungi {ital Ceratocystis piceae} and {ital Stereum sanguinolentum}, and the moulds {ital Paecilomyces variotii} and {ital Trichoderma viride} also grew with creosote. To prepare samples for IR-measurements, continuous extraction of creosote from the nutrient liquid by percolation with methylene chloride was suitable. However, the IR-spectra of creosote did not show any measurable changes after incubation with 16 bacterial strains and 6 fungi. 42 refs., 2 figs., 4 tabs.

An optimization study of solid-state fermentation: xanthophylls extraction from marigold flowers.

Science.gov (United States)

Luis, Navarrete-Bolaños José; Hugo, Jiménez-Islas; Enrique, Botello-Alvarez; Ramiro, Rico-Martínez; Octavio, Paredes-López

2004-09-01

Marigold flowers are the main natural source of xanthophylls, and marigold saponified extract is used as an additive in several food and pharmaceutical industries. In this work, the use of a solid-state fermentation (ensilage) process for increasing the yield of xanthophylls extracted from fermented marigold flowers was examined. The process consisted of a mixed culture of three microorganisms (Flavobacterium IIb, Acinetobacter anitratus, and Rhizopus nigricans), part of the normal microbiota associated with the marigold flower. These microorganisms had been previously isolated, and were identified as relevant for the ensilage process due to their capacity to produce cellulolytic enzymes. Based on experimental design strategies, optimum operation values were determined for aeration, moisture, agitation, and marigold-to-inoculum ratio in the proposed solid-state fermentation equipment, leading to a xanthophylls yield of 17.8-g/kg dry weight. The optimum achieved represents a 65% increase with respect to the control. HPLC analysis indicated conservation of extracted oleoresin. Based on the experimental results, interactions were identified that could be associated with the heat and mass-transfer reactions taking place within the bioreactor. The insight gained allows conditions that limit growth and metabolic activity to be avoided.

Sugarcane starch: quantitative determination and characterization

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Joelise de Alencar Figueira

2011-09-01

Full Text Available Starch is found in sugarcane as a storage polysaccharide. Starch concentrations vary widely depending on the country, variety, developmental stage, and growth conditions. The purpose of this study was to determine the starch content in different varieties of sugarcane, between May and November 2007, and some characteristics of sugarcane starch such as structure and granules size; gelatinization temperature; starch solution filterability; and susceptibility to glucoamylase, pullulanase, and commercial bacterial and fungal α-amylase enzymes. Susceptibility to debranching amylolytic isoamylase enzyme from Flavobacterium sp. was also tested. Sugarcane starch had spherical shape with a diameter of 1-3 µm. Sugarcane starch formed complexes with iodine, which showed greater absorption in the range of 540 to 620 nm. Sugarcane starch showed higher susceptibility to glucoamylase compared to that of waxy maize, cassava, and potato starch. Sugarcane starch also showed susceptibility to debranching amylolytic pullulanases similar to that of waxy rice starch. It also showed susceptibility to α-amylase from Bacillus subtilis, Bacillus licheniformis, and Aspergillus oryzae similar to that of the other tested starches producing glucose, maltose, maltotriose, maltotetraose, maltopentose and limit α- dextrin.

Antibiotic susceptibility profile of bacteria isolated from natural sources of water from rural areas of East Sikkim

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Shubra Poonia

2014-01-01

Full Text Available Background: Contamination of water, food, and environment with antibiotic-resistant bacteria poses a serious public health issue. Objective: The objective was to study the bacterial pollution of the natural sources of water in east Sikkim and to determine the antimicrobial profile of the bacterial isolates. Materials and Methods: A total of 225 samples, 75 each during winter, summer, and monsoon season were collected from the same source in every season for bacteriological analysis by membrane filtration method. Antibiotic susceptibility test was performed using standard disc diffusion method. Results: A total of 19 bacterial species of the genera Escherichia, Klebsiella, Proteus, Salmonella, Shigella, Enterobacter, Citrobacter, Morganella, Pseudomonas, Acinetobacter, Flavobacterium, and Serratia were isolated and their antimicrobial sensitivity tested. Generally, most bacterial isolates except Salmonella and Shigella species were found resistant to commonly used antibiotics such as ampicillin (57.5%, trimethoprim/sulfamethoxaole (39.1%, amoxicillin/clavulanic acid (37.4%, cefixime (34.5%, tetracycline (29.1%, ceftazidime (26.3%, ofloxacin (25.9%, amikacin (8.7%, and gentamicin (2.7% but sensitive to imipenem and piperacillin/tazobactam. Conclusion: Natural sources of water in east Sikkim are grossly contaminated with bacteria including enteropathogens. The consumption of untreated water from these sources might pose health risk to consumers.

Monitoring the microbial community during solid-state acetic acid fermentation of Zhenjiang aromatic vinegar.

Science.gov (United States)

Xu, Wei; Huang, Zhiyong; Zhang, Xiaojun; Li, Qi; Lu, Zhenming; Shi, Jinsong; Xu, Zhenghong; Ma, Yanhe

2011-09-01

Zhenjiang aromatic vinegar is one of the most famous Chinese traditional vinegars. In this study, change of the microbial community during its fermentation process was investigated. DGGE results showed that microbial community was comparatively stable, and the diversity has a disciplinary series of changes during the fermentation process. It was suggested that domestication of microbes and unique cycle-inoculation style used in the fermentation of Zhenjiang aromatic vinegar were responsible for comparatively stable of the microbial community. Furthermore, two clone libraries were constructed. The results showed that bacteria presented in the fermentation belonged to genus Lactobacillus, Acetobacter, Gluconacetobacter, Staphylococcus, Enterobacter, Pseudomonas, Flavobacterium and Sinorhizobium, while the fungi were genus Saccharomyces. DGGE combined with clone library analysis was an effective and credible technique for analyzing the microbial community during the fermentation process of Zhenjiang aromatic vinegar. Real-time PCR results suggested that the biomass showed a "system microbes self-domestication" process in the first 5 days, then reached a higher level at the 7th day before gradually decreasing until the fermentation ended at the 20th day. This is the first report to study the changes of microbial community during fermentation process of Chinese traditional solid-state fermentation of vinegar. Copyright © 2011 Elsevier Ltd. All rights reserved.

Microorganism selection and biosurfactant production in a continuously and periodically operated bioslurry reactor.

Science.gov (United States)

Cassidy, D P; Hudak, A J

2001-06-29

A continuous-flow reactor (CSTR) and a soil slurry-sequencing batch reactor (SS-SBR) were maintained in 8l vessels for 180 days to treat a soil contaminated with diesel fuel (DF). Concentrations of Candida tropicalis, Brevibacterium casei, Flavobacterium aquatile, Pseudomonas aeruginosa, and Pseudomonas fluorescens were determined using fatty acid methyl ester (FAME) analysis. DF removal (biological and volatile) and biosurfactant concentrations were measured. The SS-SBR encouraged the growth of biosurfactant-producing species relative to the CSTR. Counts of biosurfactant-producing species (C. tropicalis, P. aeruginosa, P. fluorescens) relative to total microbial counts were 88% in the SS-SBR and 23% in the CSTR. Biosurfactants were produced in the SS-SBR to levels of nearly 70 times the critical micelle concentration (CMC) early in the cycle, but were completely degraded by the end of each cycle. No biosurfactant production was observed in the CSTR. DF biodegradation rates were over 40% greater and DF stripping was over five times lower in the SS-SBR than the CSTR. However, considerable foaming occurred in the SS-SBR. Reversing the mode of operation in the reactors on day 80 caused a complete reversal in microbial consortia and reactor performance by day 120. These results show that bioslurry reactor operation can be manipulated to control overall reactor performance.

Impact of feed additives on surface mucosal health and columnaris susceptibility in channel catfish fingerlings, Ictalurus punctatus.

Science.gov (United States)

Zhao, Honggang; Li, Chao; Beck, Benjamin H; Zhang, Ran; Thongda, Wilawan; Davis, D Allen; Peatman, Eric

2015-10-01

One of the highest priority areas for improvement in aquaculture is the development of dietary additives and formulations which provide for complete mucosal health and protection of fish raised in intensive systems. Far greater attention has been paid to dietary impact on gut health than to protective effects at other mucosal surfaces such as skin and gill. These exterior surfaces, however, are important primary targets for pathogen attachment and invasion. Flavobacterium columnare, the causative agent of columnaris disease, is among the most prevalent of all freshwater disease-causing bacteria, impacting global aquaculture of catfish, salmonids, baitfish and aquaria-trade species among others. This study evaluated whether the feeding of a standard catfish diet supplemented with Alltech dietary additives Actigen(®), a concentrated source of yeast cell wall-derived material and/or Allzyme(®) SSF, a fermented strain of Aspergillus niger, could offer protection against F. columnare mortality. A nine-week feeding trial of channel catfish fingerlings with basal diet (B), B + Allzyme(®) SSF, B + Actigen(®) and B + Actigen(®)+Allzyme(®) SSF revealed good growth in all conditions (FCR additives may provide protection extending beyond the gut to surface mucosa. Copyright © 2015 Elsevier Ltd. All rights reserved.

Pyrosequencing of the bacteria associated with Platygyra carnosus corals with skeletal growth anomalies reveals differences in bacterial community composition in apparently healthy and diseased tissues

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Jenny Chun-Yee Ng

2015-10-01

Full Text Available Corals are rapidly declining globally due to coral diseases. Skeletal growth anomalies (SGA or coral tumors are a group of coral diseases that affect coral reefs worldwide, including Hong Kong waters in the Indo-Pacific region. To better understand how bacterial communities may vary in corals with SGA, for the first time, we examined the bacterial composition associated with the apparently healthy and the diseased tissues of SGA-affected Platgyra carnosus using 16S ribosomal rRNA gene pyrosequencing. Taxonomic analysis revealed Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria as the main phyla in both the apparently healthy and the diseased tissues. A significant difference in the bacterial community composition was observed between the two conditions at the OTU level. Diseased tissues were associated with higher abundances of Acidobacteria and Gemmatimonadetes, and a lower abundance of Spirochaetes. Several OTUs belonging to Rhodobacteraceae, Rhizobiales, Gammaproteobacteria, and Cytophaga-Flavobacterium-Bacteroidetes (CFB were strongly associated with the diseased tissues. These groups of bacteria may contain potential pathogens involved with the development of SGA or opportunistic secondary or tertiary colonizers that proliferated upon the health-compromised coral host. We suggest that these bacterial groups to be further studied based on inoculation experiments and testing of Koch’s postulates in efforts to understand the etiology and progression of SGA.

Bacterial communities in an ultrapure water containing storage tank of a power plant.

Science.gov (United States)

Bohus, Veronika; Kéki, Zsuzsa; Márialigeti, Károly; Baranyi, Krisztián; Patek, Gábor; Schunk, János; Tóth, Erika M

2011-12-01

Ultrapure waters (UPWs) containing low levels of organic and inorganic compounds provide extreme environment. On contrary to that microbes occur in such waters and form biofilms on surfaces, thus may induce corrosion processes in many industrial applications. In our study, refined saltless water (UPW) produced for the boiler of a Hungarian power plant was examined before and after storage (sampling the inlet [TKE] and outlet [TKU] waters of a storage tank) with cultivation and culture independent methods. Our results showed increased CFU and direct cell counts after the storage. Cultivation results showed the dominance of aerobic, chemoorganotrophic α-Proteobacteria in both samples. In case of TKU sample, a more complex bacterial community structure could be detected. The applied molecular method (T-RFLP) indicated the presence of a complex microbial community structure with changes in the taxon composition: while in the inlet water sample (TKE) α-Proteobacteria (Sphingomonas sp., Novosphingobium hassiacum) dominated, in the outlet water sample (TKU) the bacterial community shifted towards the dominance of α-Proteobacteria (Rhodoferax sp., Polynucleobacter sp., Sterolibacter sp.), CFB (Bacteroidetes, formerly Cytophaga-Flavobacterium-Bacteroides group) and Firmicutes. This shift to the direction of fermentative communities suggests that storage could help the development of communities with an increased tendency toward corrosion.

Biodegradability of bacterial surfactants.

Science.gov (United States)

Lima, Tânia M S; Procópio, Lorena C; Brandão, Felipe D; Carvalho, André M X; Tótola, Marcos R; Borges, Arnaldo C

2011-06-01

This work aimed at evaluating the biodegradability of different bacterial surfactants in liquid medium and in soil microcosms. The biodegradability of biosurfactants by pure and mixed bacterial cultures was evaluated through CO(2) evolution. Three bacterial strains, Acinetobacter baumanni LBBMA ES11, Acinetobacter haemolyticus LBBMA 53 and Pseudomonas sp. LBBMA 101B, used the biosurfactants produced by Bacillus sp. LBBMA 111A (mixed lipopeptide), Bacillus subtilis LBBMA 155 (lipopeptide), Flavobacterium sp. LBBMA 168 (mixture of flavolipids), Dietzia Maris LBBMA 191(glycolipid) and Arthrobacter oxydans LBBMA 201(lipopeptide) as carbon sources in minimal medium. The synthetic surfactant sodium dodecyl sulfate (SDS) was also mineralized by these microorganisms, but at a lower rate. CO(2) emitted by a mixed bacterial culture in soil microcosms with biosurfactants was higher than in the microcosm containing SDS. Biosurfactant mineralization in soil was confirmed by the increase in surface tension of the soil aqueous extracts after incubation with the mixed bacterial culture. It can be concluded that, in terms of biodegradability and environmental security, these compounds are more suitable for applications in remediation technologies in comparison to synthetic surfactants. However, more information is needed on structure of biosurfactants, their interaction with soil and contaminants and scale up and cost for biosurfactant production.

PENGOLAHAN LIMBAH TEKSTIL SISTEM KOMBINASI ANAEROBIK-AEROBIK MENGGUNAKAN BIOFILM BAKTERI KONSORSIUM DARI LUMPUR LIMBAH TEKSTIL

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I Dewa K. Sastrawidana

2012-11-01

Full Text Available The objective of this research are to assess the eficiency of biodegradation of textile dyes by using bacteriaconsortium biofim on combined anaerobic-aerobic system and also to asses the possibility for applicating this technologyin textile dyeing industry. Textile wastewater that used in this research are azo dyes total concentration 200 mg/L which isconsist of remazol yellow, remazol red, remazol black and remazol blue For anaerobik condition step, by using bacteriaconsortium selected from sludge of textile wastewater plant of Mama & Leon consist of Aeromonas sp ML6., Aeromonassp. ML14, Aeromonas sp. ML24, Pseudomonas sp. ML8 and Flavobacterium Sp ML20 whereas aerobic condition step,using bacteria consortium selected from Badung river sludge consist of Vibrio sp. and Plesiomonas sp. Both bacteriaconsortium is immobilized separatively on volcanous stone for 3 days in each reactor before used to treatment of synteticdyeswater (SDW. Further more, immobilized bacteria consortiom on volcanous stone is determined by four plate countmethod.The result ot this research showed the eficiency process in decreasing color, TDS, TSS, COD and BOD was96,94%,75,73%, 68,03%, 97,68% dan 94,60% respectively. So, wastewater treatment by using combined anaerobicaerobicsystem is potential to applied on wastewater treatment in textile industry.

Microbial evaluation and occurrence of antidrug multi-resistant organisms among the indigenous Clarias species in River Oluwa, Nigeria

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T.A. Ayandiran

2017-01-01

Full Text Available Fish may harbor pathogens on or inside its body when in contaminated environment. Clarias gariepinus and Clarias buthopogon were analyzed to evaluate the likely impact of pollution on the antidrug resistance pattern of their microbial isolates. Different bacterial and fungal counts were observed on the fish organs (skin, muscles and gills. The highest bacterial count was 1,040,000 Cfu/mL while the lowest was 101 Cfu/mL. The highest fungal count obtained was 344,000 Cfu/mL while the lowest was 65 Cfu/mL. Bacterial isolates belonging to genera Bacillus, Clostridium, Alcaligenes, Flavobacterium, Enterobacter and Corynebacterium were obtained from the organs. Also, fungal isolates belonging to the genera Penicillium, Aspergillus, Rhizopus, Monila and Fusarium were isolated. The resistance of isolates from C. gariepinus to drugs was between 50% and 90% with Bacillus species showing the highest resistance. For isolates from C. buthopogon, 40–90% resistance was observed with Alcaligenes faecalis showing highest resistance. Five patterns of multiple drug resistance were observed among the bacterial isolates with antibiotics ranging from 4 to 9. Also, result of fungal isolates showed susceptibility to ketoconazole and resistant to fluconazole and griseofulvin. The public health implications of consuming these fishes are discussed.

Microencapsulation of a putative probiotic Enterobacter species, C6-6, to protect rainbow trout, Oncorhynchus mykiss (Walbaum), against bacterial coldwater disease.

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Ghosh, B; Cain, K D; Nowak, B F; Bridle, A R

2016-01-01

Flavobacterium psychrophilum is the causative agent of bacterial coldwater disease (BCWD), which has a major impact on salmonid aquaculture globally. An Enterobacter species, C6-6, isolated from the gut of rainbow trout, Oncorhynchus mykiss (Walbaum), has been identified as a potential probiotic species providing protection against BCWD. This study examined the effects of alginate microencapsulation on the protective efficacy of C6-6 against BCWD in vivo when administered to rainbow trout fry orally or by intraperitoneal (IP) injection. Viable C6-6 bacteria were microencapsulated successfully, and this process (microencapsulation) did not significantly deteriorate its protective properties as compared to the administration of non-microencapsulated C6-6 bacteria. Both oral and IP delivery of C6-6 achieved significantly better protection than control treatments that did not contain C6-6 bacteria. The highest relative percent survival (RPS) resulted from IP delivery (71.4%) and was significantly greater than the highest oral RPS (38.6%). Successful intestinal colonization was not critical to protective effects of C6-6. The study showed that C6-6 administration, with or without encapsulation, was a viable choice for protecting fry from BCWD especially when administered intraperitoneally. © 2014 John Wiley & Sons Ltd.

The immune response of rainbow trout to Flavobacterium psychrophilum following immersion-challenge model with and without hydrogen peroxide pre-treatment

DEFF Research Database (Denmark)

Henriksen, Maya Maria Mihályi; Madsen, Lone; Kania, P. W.

2013-01-01

, no commercial vaccine is currently available and the disease is treated with antibiotics. Injection-based challenges with F. psychrophilum are standardized but the route of infection does not reflect a natural situation. Therefore, we established an immersion-based model investigating if hydrogen peroxide (H2O2...... in the gill tissue were evaluated using hematoxylin and eosin stained tissue sections. Exposure to both H2O2 and F. psychrophilum intensified tissue damage and postponed healing. The results indicate that F. psychrophilum may have an immunosuppressive action and that environmental stress may be one of several...

Flavobacterium branchiophilum and F. succinicans associated with bacterial gill disease in rainbow trout Oncorhynchus mykiss (Walbaum) in water recirculation aquaculture systems

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Raised rainbow trout Oncorhynchus mykiss in six replicated water recirculation aquaculture systems (WRAS), and manipulated environmental conditions to promote bacterial gill disease (BGD). For each episode of BGD, gill tissue was sampling from affected fish, unaffected fish within the same WRAS, and...

Bacteria-mediated arsenic oxidation and reduction in the growth media of arsenic hyperaccumulator Pteris vittata.

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Wang, Xin; Rathinasabapathi, Bala; de Oliveira, Letuzia Maria; Guilherme, Luiz R G; Ma, Lena Q

2012-10-16

Microbes play an important role in arsenic transformation and cycling in the environment. Microbial arsenic oxidation and reduction were demonstrated in the growth media of arsenic hyperaccumulator Pteris vittata L. All arsenite (AsIII) at 0.1 mM in the media was oxidized after 48 h incubation. Oxidation was largely inhibited by antibiotics, indicating that bacteria played a dominant role. To identify AsIII oxidizing bacteria, degenerate primers were used to amplify ∼500 bp of the AsIII oxidase gene aioA (aroA) using DNA extracted from the media. One aioA (aroA)-like sequence (MG-1, tentatively identified as Acinetobacter sp.) was amplified, exhibiting 82% and 91% identity in terms of gene and deduced protein sequence to those from Acinetobacter sp. 33. In addition, four bacterial strains with different arsenic tolerance were isolated and identified as Comamonas sp.C-1, Flavobacterium sp. C-2, Staphylococcus sp. C-3, and Pseudomonas sp. C-4 using carbon utilization, fatty acid profiles, and/or sequencing 16s rRNA gene. These isolates exhibited dual capacity for both AsV reduction and AsIII oxidation under ambient conditions. Arsenic-resistant bacteria with strong AsIII oxidizing ability may have potential to improve bioremediation of AsIII-contaminated water using P. vittata and/or other biochemical strategies.

Aerobic Sludge Granulation in a Full-Scale Sequencing Batch Reactor

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Jun Li

2014-01-01

Full Text Available Aerobic granulation of activated sludge was successfully achieved in a full-scale sequencing batch reactor (SBR with 50,000 m3 d−1 for treating a town’s wastewater. After operation for 337 days, in this full-scale SBR, aerobic granules with an average SVI30 of 47.1 mL g−1, diameter of 0.5 mm, and settling velocity of 42 m h−1 were obtained. Compared to an anaerobic/oxic plug flow (A/O reactor and an oxidation ditch (OD being operated in this wastewater treatment plant, the sludge from full-scale SBR has more compact structure and excellent settling ability. Denaturing gradient gel electrophoresis (DGGE analysis indicated that Flavobacterium sp., uncultured beta proteobacterium, uncultured Aquabacterium sp., and uncultured Leptothrix sp. were just dominant in SBR, whereas uncultured bacteroidetes were only found in A/O and OD. Three kinds of sludge had a high content of protein in extracellular polymeric substances (EPS. X-ray fluorescence (XRF analysis revealed that metal ions and some inorganics from raw wastewater precipitated in sludge acted as core to enhance granulation. Raw wastewater characteristics had a positive effect on the granule formation, but the SBR mode operating with periodic feast-famine, shorter settling time, and no return sludge pump played a crucial role in aerobic sludge granulation.

Selection of electrogenic bacteria for microbial fuel cell in removing Victoria blue R from wastewater.

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Chen, Chih-Yu; Tsai, Teh-Hua; Wu, Pei-Ssu; Tsao, Shuo-En; Huang, Yu-Shan; Chung, Ying-Chien

2018-01-28

This study was conducted to select electrogenic bacteria from wastewater sludge. Phylogenetic analysis revealed that Proteobacteria was the dominant phylum in the microbial fuel cell (MFC) during the decomposition process of organic pollutants. Five culturable bacteria strains - namely, Bacillus subtilis, Flavobacterium sp., Aeromonas hydrophila, Citrobacter freundii, and Stenotrophomonas sp. - have a double potential in dye removal and electricity generation. We inoculated the mixed electrogenic bacteria at a specific ratio and treated them with a triphenylmethane dye, Victoria blue R (VBR), to evaluate their electricity generation ability for the artificial and real wastewater. The results of the VBR shock-loading experiment indicated that the inoculated MFC could adapt to shock loading in 1-2 days and exhibited high removal efficiency (95-100%) for 100-800 mg L -1 VBR with a power density of 8.62 ± 0.10 to 34.81 ± 0.25 mW m -2 . The selected electrogenic bacteria in the MFC could use VBR as only electron donor for power generation. The matrix effects of the real wastewater on VBR removal and electricity generation of MFC were insignificant. VBR degradation by the electrogenic bacteria involves a stepwise demethylation process to yield partially dealkylated VBR species. In addition, these results demonstrate the feasibility of inoculating culturable bacteria strains to develop an efficient MFC for purifying wastewater.

Microbial ecology of Thailand tsunami and non-tsunami affected terrestrials.

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Somboonna, Naraporn; Wilantho, Alisa; Jankaew, Kruawun; Assawamakin, Anunchai; Sangsrakru, Duangjai; Tangphatsornruang, Sithichoke; Tongsima, Sissades

2014-01-01

The effects of tsunamis on microbial ecologies have been ill-defined, especially in Phang Nga province, Thailand. This ecosystem was catastrophically impacted by the 2004 Indian Ocean tsunami as well as the 600 year-old tsunami in Phra Thong island, Phang Nga province. No study has been conducted to elucidate their effects on microbial ecology. This study represents the first to elucidate their effects on microbial ecology. We utilized metagenomics with 16S and 18S rDNA-barcoded pyrosequencing to obtain prokaryotic and eukaryotic profiles for this terrestrial site, tsunami affected (S1), as well as a parallel unaffected terrestrial site, non-tsunami affected (S2). S1 demonstrated unique microbial community patterns than S2. The dendrogram constructed using the prokaryotic profiles supported the unique S1 microbial communities. S1 contained more proportions of archaea and bacteria domains, specifically species belonging to Bacteroidetes became more frequent, in replacing of the other typical floras like Proteobacteria, Acidobacteria and Basidiomycota. Pathogenic microbes, including Acinetobacter haemolyticus, Flavobacterium spp. and Photobacterium spp., were also found frequently in S1. Furthermore, different metabolic potentials highlighted this microbial community change could impact the functional ecology of the site. Moreover, the habitat prediction based on percent of species indicators for marine, brackish, freshwater and terrestrial niches pointed the S1 to largely comprise marine habitat indicating-species.

Citric acid application for denitrification process support in biofilm reactor.

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Mielcarek, Artur; Rodziewicz, Joanna; Janczukowicz, Wojciech; Dabrowska, Dorota; Ciesielski, Slawomir; Thornton, Arthur; Struk-Sokołowska, Joanna

2017-03-01

The study demonstrated that citric acid, as an organic carbon source, can improve denitrification in Anaerobic Sequencing Batch Biofilm Reactor (AnSBBR). The consumption rate of the organic substrate and the denitrification rate were lower during the period of the reactor's acclimatization (cycles 1-60; 71.5 mgCOD L -1  h -1 and 17.81 mgN L -1  h -1 , respectively) than under the steady state conditions (cycles 61-180; 143.8 mgCOD L -1  h -1 and 24.38 mgN L -1  h -1 ). The biomass yield coefficient reached 0.04 ± 0.02 mgTSS· mgCOD re -1 (0.22 ± 0.09 mgTSS mgN re -1 ). Observations revealed the diversified microbiological ecology of the denitrifying bacteria. Citric acid was used mainly by bacteria representing the Trichoccocus genus, which represented above 40% of the sample during the first phase of the process (cycles 1-60). In the second phase (cycles 61-180) the microorganisms the genera that consumed the acetate and formate, as the result of citric acid decomposition were Propionibacterium (5.74%), Agrobacterium (5.23%), Flavobacterium (1.32%), Sphaerotilus (1.35%), Erysipelothrix (1.08%). Copyright © 2016 Elsevier Ltd. All rights reserved.

Phytoremediation of mercury in pristine and crude oil contaminated soils: Contributions of rhizobacteria and their host plants to mercury removal.

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Sorkhoh, N A; Ali, N; Al-Awadhi, H; Dashti, N; Al-Mailem, D M; Eliyas, M; Radwan, S S

2010-11-01

The rhizospheric soils of three tested legume crops: broad beans (Vicia faba), beans (Phaseolus vulgaris) and pea (Pisum sativum), and two nonlegume crops: cucumber (Cucumis sativus) and tomato, (Lycopersicon esculentum) contained considerable numbers (the magnitude of 10(5)g(-1) soil) of bacteria with the combined potential for hydrocarbon-utilization and mercury-resistance. Sequencing of the 16S rRNA coding genes of rhizobacteria associated with broad beans revealed that they were affiliated to Citrobacter freundii, Enterobacter aerogenes, Exiquobacterium aurantiacum, Pseudomonas veronii, Micrococcus luteus, Brevibacillus brevis, Arthrobacter sp. and Flavobacterium psychrophilum. These rhizobacteria were also diazotrophic, i.e. capable of N(2) fixation, which makes them self-sufficient regarding their nitrogen nutrition and thus suitable remediation agents in nitrogen-poor soils, such as the oily desert soil. The crude oil attenuation potential of the individual rhizobacteria was inhibited by HgCl(2), but about 50% or more of this potential was still maintained in the presence of up to 40 mgl(-1) HgCl(2). Rhizobacteria-free plants removed amounts of mercury from the surrounding media almost equivalent to those removed by the rhizospheric bacterial consortia in the absence of the plants. It was concluded that both the collector plants and their rhizospheric bacterial consortia contributed equivalently to mercury removal from soil. Copyright © 2010 Elsevier Inc. All rights reserved.

Entericidin is required for a probiotic treatment (Enterobacter sp. strain C6-6) to protect trout from cold-water disease challenge.

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Schubiger, Carla B; Orfe, Lisa H; Sudheesh, Ponnerassery S; Cain, Kenneth D; Shah, Devendra H; Call, Douglas R

2015-01-01

Flavobacterium psychrophilum causes bacterial cold-water disease in multiple fish species, including salmonids. An autochthonous Enterobacter strain (C6-6) inhibits the in vitro growth of F. psychrophilum, and when ingested as a putative probiotic, it provides protection against injection challenge with F. psychrophilum in rainbow trout. In this study, low-molecular-mass (≤3 kDa) fractions from both Enterobacter C6-6 and Escherichia coli K-12 culture supernatants inhibited the growth of F. psychrophilum. The ≤3-kDa fraction from Enterobacter C6-6 was analyzed by SDS-PAGE, and subsequent tandem mass spectroscopy identified EcnB, which is a small membrane lipoprotein that is a putative pore-forming toxin. Agar plate diffusion assays demonstrated that ecnAB knockout strains of both Enterobacter C6-6 and E. coli K-12 no longer inhibited F. psychrophilum (P ) and the wild-type strain (C6-6) were added to the fish diet every day for 38 days. On day 11, the fish were challenged by injection with a virulent strain of F. psychrophilum (CSF 259-93). Fish that were fed C6-6 had significantly longer survival than fish fed the ecnAB knockout strain (P Enterobacter C6-6, and it may present new opportunities for therapeutic and prophylactic treatments against similarly susceptible pathogens. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Treatment of Chryseobacterium indologenes ventilator-associated pneumonia in a critically ill trauma patient.

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Monteen, Megan R; Ponnapula, Supriya; Wood, G Christopher; Croce, Martin A; Swanson, Joseph M; Boucher, Bradley A; Fabian, Timothy C

2013-12-01

To report a case of Chryseobacterium indologenes ventilator-associated pneumonia (VAP) in a critically ill trauma patient. This report describes a 66-year-old critically ill trauma patient who developed VAP, which was caused by C indologenes. The patient was injured in a riding lawn mower accident that trapped him underwater in a pond. The patient required surgery for intra-abdominal injuries and was mechanically ventilated in the trauma intensive care unit. On hospital day 5, the patient developed signs and symptoms of VAP. A diagnosis of C indologenes VAP was confirmed based on a quantitative culture from a bronchoscopic bronchoalveolar lavage. The patient's infection was successfully treated with moxifloxacin for 2 days followed by cefepime for 7 days. Formally known as Flavobacterium indologenes, C indologenes is a Gram-negative bacillus normally found in plants, soil, foodstuffs, and fresh and marine water sources. Recently, worldwide reports of C indologenes infections in humans have been increasing, though reports from the United States are still rare. Bacteremia and pneumonia are the most commonly reported infections, and most patients are immunocompromised. The current case differs from most previous reports because this patient was in the United States and did not have any traditional immunocompromised states (eg, transplant, cancer, HIV/AIDS, or corticosteroid use). This case report demonstrates that C indologenes can cause VAP in a trauma ICU patient.

The identification of genes specific to Prevotella intermedia and Prevotella nigrescens using genomic subtractive hybridization.

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Masakiyo, Yoshiaki; Yoshida, Akihiro; Shintani, Yasuyuki; Takahashi, Yusuke; Ansai, Toshihiro; Takehara, Tadamichi

2010-06-01

Prevotella intermedia and Prevotella nigrescens, which are often isolated from periodontal sites, were once considered two different genotypes of P. intermedia. Although the genomic sequence of P. intermedia was determined recently, little is known about the genetic differences between P. intermedia and P. nigrescens. The subtractive hybridization technique is a powerful method for generating a set of DNA fragments differing between two closely related bacterial strains or species. We used subtractive hybridization to identify the DNA regions specific to P. intermedia ATCC 25611 and P. nigrescens ATCC 25261. Using this method, four P. intermedia ATCC 25611-specific and three P. nigrescens ATCC 25261-specific regions were determined. From the species-specific regions, insertion sequence (IS) elements were isolated for P. intermedia. IS elements play an important role in the pathogenicity of bacteria. For the P. intermedia-specific regions, the genes adenine-specific DNA-methyltransferase and 8-amino-7-oxononanoate synthase were isolated. The P. nigrescens-specific region contained a Flavobacterium psychrophilum SprA homologue, a cell-surface protein involved in gliding motility, Prevotella melaninogenica ATCC 25845 glutathione peroxide, and Porphyromonas gingivalis ATCC 33277 leucyl-tRNA synthetase. The results demonstrate that the subtractive hybridization technique was useful for distinguishing between the two closely related species. Furthermore, this technique will contribute to our understanding of the virulence of these species. 2009 Elsevier Ltd. All rights reserved.

Combination of upflow anaerobic sludge blanket (UASB) and membrane bioreactor (MBR) for berberine reduction from wastewater and the effects of berberine on bacterial community dynamics.

Science.gov (United States)

Qiu, Guanglei; Song, Yonghui; Zeng, Ping; Duan, Liang; Xiao, Shuhu

2013-02-15

Berberine is a broad-spectrum antibiotic extensively used in personal medication. The production of berberine results in the generation of wastewater containing concentrated residual berberine. However, few related studies up to date focus on berberine removal from wastewaters. In this study, a lab-scale upflow anaerobic sludge blanket (UASB)-membrane bioreactor (MBR) process was developed for berberine removal from synthetic wastewater. The performance of the UASB-MBR system on berberine, COD and NH(4)(+)--N removal was investigated at different berberine loadings. And the effects of berberine on bacterial communities were evaluated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Results showed that, as the increase of berberine loadings, UASB performance was affected remarkably, whereas, efficient and stable performance of MBR ensured the overall removal rates of berberine, COD and NH(4)(+)--N consistently reached up to 99%, 98% and 98%, respectively. Significant shifts of bacterial community structures were detected in both UASB and MBR, especially in the initial operations. Along with the increase of berberine loadings, high antibiotic resisting species and some functional species, i.e. Acinetobacter sp., Clostridium sp., Propionibacterium sp., and Sphingomonas sp. in UASB, as well as Sphingomonas sp., Methylocystis sp., Hydrogenophaga sp. and Flavobacterium sp. in MBR were enriched in succession. Copyright © 2013 Elsevier B.V. All rights reserved.

Prokaryotic community composition involved production of nitrogen in sediments of Mejillones Bay

International Nuclear Information System (INIS)

Moraga, Ruben; Galan, Alexander; Rosello-Mora, Ramon; Araya, Ruben; Valdes, Jorge

2014-01-01

Conventional denitrification and anaerobic ammonium oxidation (anammox) contributes to nitrogen loss in oxygen-deficient systems, thereby influencing many aspects of ecosystem function and global biogeochemistry. Mejillones Bay, northern Chile, presents ideal conditions to study nitrogen removal processes, because it is inserted in a coastal upwelling system, its sediments have anoxia and hypoxia conditions and under the influence of the Oxygen Minimum Zone (OMZ), unknown processes that occur there and what are the microbial communities responsible for their removal. Microbial communities associated with coastal sediments of Mejillones Bay were studied by denaturing gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH), by incubation experiments with 15 N isotope tracers were studied nitrogen loss processes operating in these sediments. DGGE analysis showed high bacterial diversity, certain redundant phylotypes and differences in community structure given by the depth; this reflects the microbial community adaptations to environmental conditions. A large fraction (up to 70%) of DAPI-stained cells hybridized with the bacterial probes. Nearly 52-90% of the cell could be further identified to know phyla. Members of the Cytophaga-Flavobacterium cluster were most abundant in the sediments (13-26%), followed by Proteobacteria. Isotopic tracer experiments for the sediments studied indicated that nitrogen loss processes that predominated were performed by denitrifying communities (43.31-111.20 μMd -1 ) was not possible to detect anammox in the area and not anammox bacteria were detected

Biochar in Co-Contaminated Soil Manipulates Arsenic Solubility and Microbiological Community Structure, and Promotes Organochlorine Degradation

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Gregory, Samuel J.; Anderson, Christopher W. N.; Camps-Arbestain, Marta; Biggs, Patrick J.; Ganley, Austen R. D.; O’Sullivan, Justin M.; McManus, Michael T.

2015-01-01

We examined the effect of biochar on the water-soluble arsenic (As) concentration and the extent of organochlorine degradation in a co-contaminated historic sheep-dip soil during a 180-d glasshouse incubation experiment. Soil microbial activity, bacterial community and structure diversity were also investigated. Biochar made from willow feedstock (Salix sp) was pyrolysed at 350 or 550°C and added to soil at rates of 10 g kg-1 and 20 g kg-1 (representing 30 t ha-1 and 60 t ha-1). The isomers of hexachlorocyclohexane (HCH) alpha-HCH and gamma-HCH (lindane), underwent 10-fold and 4-fold reductions in concentration as a function of biochar treatment. Biochar also resulted in a significant reduction in soil DDT levels (P biochar treatments after 60 days of treatment compared to the control. 16S amplicon sequencing revealed that biochar-amended soil contained more members of the Chryseobacterium, Flavobacterium, Dyadobacter and Pseudomonadaceae which are known bioremediators of hydrocarbons. We hypothesise that a recorded short-term reduction in the soluble As concentration due to biochar amendment allowed native soil microbial communities to overcome As-related stress. We propose that increased microbiological activity (dehydrogenase activity) due to biochar amendment was responsible for enhanced degradation of organochlorines in the soil. Biochar therefore partially overcame the co-contaminant effect of As, allowing for enhanced natural attenuation of organochlorines in soil. PMID:25923541

Antimicrobial activities of secondary metabolites and phylogenetic study of sponge endosymbiotic bacteria, Bacillus sp. at Agatti Island, Lakshadweep Archipelago

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Gopi Mohan

2016-09-01

Full Text Available Twenty-one species of sponges were recorded under the class of Demospongiae and Calcareous sponges of which 19 species were new to Agatti reef. A total of 113 Sponge endosymbiotic bacterial strains were isolated from twenty-one species of sponges and screened for antimicrobial activity. Five bacterial strains of sponge endosymbiotic bacteria (SEB namely SEB32, SEB33, SEB36, SEB43 and SEB51 showed antimicrobial activity against virulent marine fish pathogens such as Vibrio alginolyticus, Vibrio vulnificus, Vibrio parahaemolyticus, Aeromonas salmonicida, Flavobacterium sp., Edwardsiella sp., Proteus mirabilis and Citrobacter brackii. The secondary metabolites produced by SEB32 from sponge Dysidea fragilis (Montagu, 1818 [48] was selected with broad range of antibacterial activity and subjected for production, characterization by series of chromatography techniques and spectroscopic methods. Based on the results of FT-IR and mass spectrometry, the active molecule was tentatively predicted as “Pyrrol” and the structure is Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro- with molecular formula of C7H10N2O2. The LC50 of active molecule was 31 μg/ml and molecular weight of the metabolites was 154. The potential strain SEB32 was identified by gene sequence (GenBank Accession number JX985748 and identified as Bacillus sp. from GenBank database.

DISTOCIA EN UNA SERPIENTE RATONERA AMARILLA (Coelognathus flavolineatus, SCHLEGEL 1837-REPORTE DE CASO

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D. M. Estrada

2015-01-01

Full Text Available La distocia en reptiles es una patología caracterizada por la dificultad de llevar a cabo el proceso de parto u ovoposición, la literatura es escasa en lo que concierne a distocias en serpientes, por tanto, existen vacíos frente a este tema. s e describe el caso de una serpiente Coelognathus flavolineatus de 10 años de edad que presentaba dificultad en la ovoposición. Fueron evaluados parámetros sanguíneos, radiográficos y ecográficos, así como cultivo bacteriano y antibiograma, con el fin de identificar las posibles causas y complicaciones que desencadenaron el proceso de distocia en la serpiente. e n cuanto al tratamiento de la distocia, se realizó una punción aspirativa del huevo a través de la cloaca extrayéndose su contenido, posteriormente se procedió a extraer los restos y se tomaron hisopados de la mucosa del tracto reproductivo. Las muestras seleccionadas fueron sometidas a cultivo, luego de lo cual, se aislaron cepas de Salmonella del grupo F-67 y Escherichia coli , y en menor proporción cepas de Morganella morganii, Pseudomona aeruginosa, Flavobacterium y Proteus , las cuales se atribuyen como agentes infecciosos involucrados en la distocia en la serpiente.

Contaminants as viral cofactors: assessing indirect population effects

Science.gov (United States)

Springman, Katherine R.; Kurath, Gael; Anderson, James J.; Emlen, John M.

2005-01-01

Current toxicological methods often miss contaminant effects, particularly when immune suppression is involved. The failure to recognize and evaluate indirect and sublethal effects severely limits the applicability of those methods at the population level. In this study, the Vitality model is used to evaluate the population level effects of a contaminant exerting only indirect, sublethal effects at the individual level. Juvenile rainbow trout (Oncorhynchus mykiss) were injected with 2.5 or 10.0 mg/kg doses of the model CYP1A inducer, β-naphthoflavone (BNF) as a pre-stressor, then exposed to a challenge dose of 102 or 104 pfu/fish of infectious hematopoietic necrosis virus (IHNV), an important viral pathogen of salmonids in North America. At the end of the 28-d challenge, the mortality data were processed according to the Vitality model which indicated that the correlation between the average rate of vitality loss and the pre-stressor dose was strong:R2 = 0.9944. Average time to death and cumulative mortality were dependent on the BNF dose, while no significant difference between the two viral dosages was shown, implying that the history of the organism at the time of stressor exposure is an important factor in determining the virulence or toxicity of the stressor. The conceptual framework of this model permits a smoother transfer of results to a more complex stratum, namely the population level, which allows the immunosuppressive results generated by doses of a CYP1A inducer that more accurately represent the effects elicited by environmentally-relevant contaminant concentrations to be extrapolated to target populations. The indirect effects of other environmental contaminants with similar biotransformation pathways, such as polycyclic aromatic hydrocarbons (PAH), could be assessed and quantified with this model and the results applied to a more complex biological hierarchy.

Detection and Quantification of Flavobacterium psychrophilum-Specific Bacteriophages In Vivo in Rainbow Trout upon Oral Administration: Implications for Disease Control in Aquaculture

DEFF Research Database (Denmark)

Christiansen, Rói Hammershaimb; Dalsgaard, Inger; Middelboe, Mathias

2014-01-01

different methods-bath, oral intubation into the stomach, and phage-coated feed-with special emphasis on the oral route of delivery. Phages could be detected in all the organs investigated (intestine, spleen, brain, and kidney) 0.5 h postadministration, reaching concentrations as high as ∼10(5) PFU mg...... intestine(-1) and ∼10(3) PFU mg spleen(-1) within the first 24 h following the bath and ∼10(7) PFU mg intestine(-1) and ∼10(4) PFU mg spleen(-1) within the first 24 h following oral intubation. The phages were most persistent in the organs for the first 24 h and then decreased exponentially; no phages were...... detected after 83 h in the organs investigated. Phage administration via feed resulted in the detection of phages in the intestine, spleen, and kidney 1 h after feeding. Average concentrations of ∼10(4) PFU mg intestine(-1) and ∼10(1) PFU mg spleen(-1) were found throughout the experimental period (200 h...

Bacterial networks and co-occurrence relationships in the lettuce root microbiota.

Science.gov (United States)

Cardinale, Massimiliano; Grube, Martin; Erlacher, Armin; Quehenberger, Julian; Berg, Gabriele

2015-01-01

Lettuce is one of the most common raw foods worldwide, but occasionally also involved in pathogen outbreaks. To understand the correlative structure of the bacterial community as a network, we studied root microbiota of eight ancient and modern Lactuca sativa cultivars and the wild ancestor Lactuca serriola by pyrosequencing of 16S rRNA gene amplicon libraries. The lettuce microbiota was dominated by Proteobacteria and Bacteriodetes, as well as abundant Chloroflexi and Actinobacteria. Cultivar specificity comprised 12.5% of the species. Diversity indices were not different between lettuce cultivar groups but higher than in L. serriola, suggesting that domestication lead to bacterial diversification in lettuce root system. Spearman correlations between operational taxonomic units (OTUs) showed that co-occurrence prevailed over co-exclusion, and complementary fluorescence in situ hybridization-confocal laser scanning microscopy (FISH-CLSM) analyses revealed that this pattern results from both potential interactions and habitat sharing. Predominant taxa, such as Pseudomonas, Flavobacterium and Sphingomonadaceae rather suggested interactions, even though these are not necessarily part of significant modules in the co-occurrence networks. Without any need for complex interactions, single organisms are able to invade into this microbial network and to colonize lettuce plants, a fact that can influence the susceptibility to pathogens. The approach to combine co-occurrence analysis and FISH-CLSM allows reliably reconstructing and interpreting microbial interaction networks. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

Microbial Diversity of a Heavily Polluted Microbial Mat and Its Community Changes following Degradation of Petroleum Compounds

Science.gov (United States)

Abed, Raeid M. M.; Safi, Nimer M. D.; Köster, Jürgen; de Beer, Dirk; El-Nahhal, Yasser; Rullkötter, Jürgen; Garcia-Pichel, Ferran

2002-01-01

We studied the microbial diversity of benthic cyanobacterial mats inhabiting a heavily polluted site in a coastal stream (Wadi Gaza) and monitored the microbial community response induced by exposure to and degradation of four model petroleum compounds in the laboratory. Phormidium- and Oscillatoria-like cyanobacterial morphotypes were dominant in the field. Bacteria belonging to different groups, mainly the Cytophaga-Flavobacterium-Bacteriodes group, the γ and β subclasses of the class Proteobacteria, and the green nonsulfur bacteria, were also detected. In slurry experiments, these communities efficiently degraded phenanthrene and dibenzothiophene completely in 7 days both in the light and in the dark. n-Octadecane and pristane were degraded to 25 and 34% of their original levels, respectively, within 7 days, but there was no further degradation until 40 days. Both cyanobacterial and bacterial communities exhibited noticeable changes concomitant with degradation of the compounds. The populations enriched by exposure to petroleum compounds included a cyanobacterium affiliated phylogenetically with Halomicronema. Bacteria enriched both in the light and in the dark, but not bacteria enriched in any of the controls, belonged to the newly described Holophaga-Geothrix-Acidobacterium phylum. In addition, another bacterial population, found to be a member of green nonsulfur bacteria, was detected only in the bacteria treated in the light. All or some of the populations may play a significant role in metabolizing the petroleum compounds. We concluded that the microbial mats from Wadi Gaza are rich in microorganisms with high biodegradative potential. PMID:11916684

Determination of physiological, taxonomic, and molecular characteristics of a cultivable arsenic-resistant bacterial community.

Science.gov (United States)

Cordi, A; Pagnout, C; Devin, S; Poirel, J; Billard, P; Dollard, M A; Bauda, P

2015-09-01

A collection of 219 bacterial arsenic-resistant isolates was constituted from neutral arsenic mine drainage sediments. Isolates were grown aerobically or anaerobically during 21 days on solid DR2A medium using agar or gelan gum as gelling agent, with 7 mM As(III) or 20 mM As(V) as selective pressure. Interestingly, the sum of the different incubation conditions used (arsenic form, gelling agent, oxygen pressure) results in an overall increase of the isolate diversity. Isolated strains mainly belonged to Proteobacteria (63%), Actinobacteria (25%), and Bacteroidetes (10%). The most representative genera were Pseudomonas (20%), Acinetobacter (8%), and Serratia (15%) among the Proteobacteria; Rhodococcus (13%) and Microbacterium (5%) among Actinobacteria; and Flavobacterium (13%) among the Bacteroidetes. Isolates were screened for the presence of arsenic-related genes (arsB, ACR3(1), ACR3(2), aioA, arsM, and arrA). In this way, 106 ACR3(1)-, 74 arsB-, 22 aioA-, 14 ACR3(2)-, and one arsM-positive PCR products were obtained and sequenced. Analysis of isolate sensitivity toward metalloids (arsenite, arsenate, and antimonite) revealed correlations between taxonomy, sensitivity, and genotype. Antimonite sensitivity correlated with the presence of ACR3(1) mainly present in Bacteroidetes and Actinobacteria, and arsenite or antimonite resistance correlated with arsB gene presence. The presence of either aioA gene or several different arsenite carrier genes did not ensure a high level of arsenic resistance in the tested conditions.

Assessment of the bacteriological activity associated with granular activated carbon treatment of drinking water.

Science.gov (United States)

Stewart, M H; Wolfe, R L; Means, E G

1990-01-01

Bacteriological analyses were performed on the effluent from a conventional water treatment pilot plant in which granular activated carbon (GAC) had been used as the final process to assess the impact of GAC on the microbial quality of the water produced. Samples were collected twice weekly for 160 days from the effluents of six GAC columns, each of which used one of four different empty-bed contact times (7.5, 15, 30, and 60 min). The samples were analyzed for heterotrophic plate counts and total coliforms. Effluent samples were also exposed to chloramines and free chlorine for 60 min (pH 8.2, 23 degrees C). Bacterial identifications were performed on the disinfected and nondisinfected effluents. Additional studies were conducted to assess the bacteriological activity associated with released GAC particles. The results indicated that heterotrophic plate counts in the effluents from all columns increased to 10(5) CFU/ml within 5 days and subsequently stabilized at 10(4) CFU/ml. The heterotrophic plate counts did not differ at different empty-bed contact times. Coliforms (identified as Enterobacter spp.) were recovered from the nondisinfected effluent on only two occasions. The disinfection results indicated that 1.5 mg of chloramines per liter inactivated approximately 50% more bacteria than did 1.0 mg of free chlorine per liter after 1 h of contact time. Chloramines and chlorine selected for the development of different bacterial species--Pseudomonas spp. and Flavobacterium spp., respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2082828

Radiation disinfection of rice-straw products

International Nuclear Information System (INIS)

Ito, Hitoshi; Ishigaki, Isao; Ohki, Yumi.

1991-01-01

For the quarantine treatment of rice-straw products from foreign countries, irradiation effects of gamma-rays and electron beams on plant pathogenic microorganisms especially on fungi were investigated. The total aerobic bacteria in rice-straw was determined to be 3x10 7 - 3x10 8 per gram which consisted mainly of Pseudomonas, Flavobacterium, Arthrobacter and Erwinia. The principal bacteria in rice-straw could be eliminated with 5 kGy of gamma irradiation. Deinococcus proteolyticus and Pseudomonas radiora were the main survivors at 5 to 12 kGy of irradiation. Saprophytic fungus which belongs to Dimorphospora also survived up to 8 kGy of irradiation. The D 10 values of 26 strains of fungi isolated from rice-straw were 1.1 to 2.5 times higher in the dry condition compared to the values when irradiated in 0.067 M phosphate buffer solution. The induction dose in the dry condition also increased from 1.5 to 10 times than that in the wet condition. In the case of electron beam irradiation of fungi under dry conditions, D 10 values were about 1.3 times higher than that of gamma irradiation. From this study, the dose necessary to reduce the plant pathogenic fungi in rice-straw at a level below 10 -4 per gram was estimated to be as 7-8 kGy for gamma-irradiation and 10 kGy for electron beam irradiation. (author)

Bacterial communities associated with biofouling materials used in bench-scale hydrocarbon bioremediation.

Science.gov (United States)

Al-Mailem, Dina; Kansour, Mayada; Radwan, Samir

2015-03-01

Biofouling material samples from the Arabian (Persian) Gulf, used as inocula in batch cultures, brought about crude oil and pure-hydrocarbon removal in a mineral medium. Without any added nitrogen fertilizers, the hydrocarbon-removal values were between about 10 and 50 %. Fertilization with NaNO3 alone or together with a mixture of the vitamins thiamine, pyridoxine, vitamin B12, biotin, riboflavin, and folic acid increased the hydrocarbon-removal values, to reach 90 %. Biofouling material samples harbored total bacteria in the magnitude of 10(7) cells g(-1), about 25 % of which were hydrocarbonoclastic. These numbers were enhanced by NaNO3 and vitamin amendment. The culture-independent analysis of the total bacterioflora revealed the predominance of the gammaproteobacterial genera Marinobacter, Acinetobacter, and Alcanivorax, the Flavobacteriia, Flavobacterium, Gaetbulibacter, and Owenweeksia, and the Alphaproteobacteria Tistrella, Zavarzinia, and others. Most of those bacteria are hydrocarbonoclastic. Culture-dependent analysis of hydrocarbonoclastic bacteria revealed that Marinobacter hydrocarbonoclasticus, Dietzia maris, and Gordonia bronchialis predominated in the fouling materials. In addition, each material had several more-specific hydrocarbonoclastic species, whose frequencies were enhanced by NaNO3 and vitamin fertilization. The same samples of fouling materials were used in four successive crude-oil-removal cycles without any dramatic loss of their hydrocarbon-removal potential nor of their associated hydrocarbonoclastic bacteria. In the fifth cycle, the oil-removal value was reduced by about 50 % in only one of the studied samples. This highlights how firmly biofouling materials were immobilizing the hydrocarbonoclastic bacteria.

Chemical composition and antimicrobial activity of Cymbopogon nardus citronella essential oil against systemic bacteria of aquatic animals

Science.gov (United States)

Wei, Lee Seong; Wee, Wendy

2013-01-01

Background & Objectives This paper describes chemical composition and antimicrobial activity of Cymbopogon nardus citronella essential oil against Edwardsiella spp. (n = 21), Vibrio spp. (n = 6), Aeromonas spp. (n = 2), Escherichia coli (n = 2), Salmonella spp. (n = 2), Flavobacterium spp. (n = 1), Pseudomonas spp. (n = 1) and Streptococcus spp. (n = 1) isolated from internal organs of aquatic animals. Due to the ban of antibiotics for aquaculture use, this study was carried out to evaluate the potential of citronella essential oil as alternative to commercial antibiotic use against systemic bacteria in cultured aquatic animals. Materials & Methods The essential oil of C. nardus was prepared by using the steam distillation method and the chemical composition of the essential oil was analyzed by gas chromatography–mass spectroscopy (GC–MS). Minimum inhibitory concentration (MIC) of the essential oil tested against bacterial isolates from various aquatic animals and ATCC type strains were determined using two-fold broth micro dilution method with kanamycin and eugenol as positive controls. Results A total of 22 chemical compounds were detected in C. nardus essential oil with 6-octenal, 3, 7-dimethyl- or citronellal representing the major compounds (29.6%). The MIC values of the citronella oil ranged from 0.244 µg/ml to 0.977 µg/ml when tested against the bacterial isolates. Conclusion The results of the present study revealed the potential of C. nardus essential oil as alternative to commercial antibiotics for aquaculture use. PMID:23825733

Phylogenetic and physiological diversity of microorganisms isolated from a deep greenland glacier ice core

Science.gov (United States)

Miteva, V. I.; Sheridan, P. P.; Brenchley, J. E.

2004-01-01

We studied a sample from the GISP 2 (Greenland Ice Sheet Project) ice core to determine the diversity and survival of microorganisms trapped in the ice at least 120,000 years ago. Previously, we examined the phylogenetic relationships among 16S ribosomal DNA (rDNA) sequences in a clone library obtained by PCR amplification from genomic DNA extracted from anaerobic enrichments. Here we report the isolation of nearly 800 aerobic organisms that were grouped by morphology and amplified rDNA restriction analysis patterns to select isolates for further study. The phylogenetic analyses of 56 representative rDNA sequences showed that the isolates belonged to four major phylogenetic groups: the high-G+C gram-positives, low-G+C gram-positives, Proteobacteria, and the Cytophaga-Flavobacterium-Bacteroides group. The most abundant and diverse isolates were within the high-G+C gram-positive cluster that had not been represented in the clone library. The Jukes-Cantor evolutionary distance matrix results suggested that at least 7 isolates represent new species within characterized genera and that 49 are different strains of known species. The isolates were further categorized based on the isolation conditions, temperature range for growth, enzyme activity, antibiotic resistance, presence of plasmids, and strain-specific genomic variations. A significant observation with implications for the development of novel and more effective cultivation methods was that preliminary incubation in anaerobic and aerobic liquid prior to plating on agar media greatly increased the recovery of CFU from the ice core sample.

Analysis of extracellular polymeric substances (EPS) and ciprofloxacin-degrading microbial community in the combined Fe-C micro-electrolysis-UBAF process for the elimination of high-level ciprofloxacin.

Science.gov (United States)

Zhang, Longlong; Yue, Qinyan; Yang, Kunlun; Zhao, Pin; Gao, Baoyu

2018-02-01

Extracellular polymeric substances (EPS) and ciprofloxacin-degrading microbial community in the combined Fe-C micro-electrolysis and up-flow biological aerated filter (UBAF) process for the treatment of high-level ciprofloxacin (CIP) were analyzed. The research demonstrated a great potential of Fe-C micro-electrolysis-UBAF for the elimination of high-level CIP. Above 90% of CIP removal was achieved through the combined process at 100 mg L -1 of CIP loading. In UBAF, the pollutants were mainly removed at 0-70 cm heights. Three-dimensional fluorescence spectrum (3D-EEM) was used to characterize the chemical structural of loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) extracted from biofilm sample in UBAF. The results showed that the protein-like substances in LB-EPS and TB-EPS had no clear change in the study. Nevertheless, an obvious release of polysaccharides in EPSs was observed during long-term exposure to CIP, which was considered as a protective response of microbial to CIP toxic. The high-throughput sequencing results revealed that the biodiversity of bacteria community became increasingly rich with gradual ciprofloxacin biodegradation in UBAF. The ciprofloxacin-degrading microbial community was mainly dominated by Proteobacteria and Bacteroidetes. Microorganisms from genera Dechloromonas, Brevundimonas, Flavobacterium, Sphingopyxis and Bosea might take a major role in ciprofloxacin degradation. This study provides deep theoretical guidance for real CIP wastewater treatment. Copyright © 2017. Published by Elsevier Ltd.

Denitrification in groundwater at uranium mill tailings sites

International Nuclear Information System (INIS)

Goering, Timothy J.; Groffman, Armando; Thomson, Bruce

1992-01-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Large scale treatment of total petroleum-hydrocarbon contaminated groundwater using bioaugmentation.

Science.gov (United States)

Poi, Gregory; Shahsavari, Esmaeil; Aburto-Medina, Arturo; Mok, Puah Chum; Ball, Andrew S

2018-05-15

Bioaugmentation or the addition of microbes to contaminated sites has been widely used to treat contaminated soil or water; however this approach is often limited to laboratory based studies. In the present study, large scale bioaugmentation has been applied to total petroleum hydrocarbons (TPH)-contaminated groundwater at a petroleum facility. Initial TPH concentrations of 1564 mg L -1 in the field were reduced to 89 mg L -1 over 32 days. This reduction was accompanied by improved ecotoxicity, as shown by Brassica rapa germination numbers that increased from 52 at day 0 to 82% by the end of the treatment. Metagenomic analysis indicated that there was a shift in the microbial community when compared to the beginning of the treatment. The microbial community was dominated by Proteobacteria and Bacteroidetes from day 0 to day 32, although differences at the genus level were observed. The predominant genera at the beginning of the treatment (day 0 just after inoculation) were Cloacibacterium, Sediminibacterium and Brevundimonas while at the end of the treatment members of Flavobacterium dominated, reaching almost half the population (41%), followed by Pseudomonas (6%) and Limnobacter (5.8%). To the author's knowledge, this is among the first studies to report the successful large scale biodegradation of TPH-contaminated groundwater (18,000 L per treatment session) at an offshore petrochemical facility. Copyright © 2018 Elsevier Ltd. All rights reserved.

Radiation disinfection of rice-straw products

Energy Technology Data Exchange (ETDEWEB)

Ito, Hitoshi; Ishigaki, Isao (Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment); Ohki, Yumi

1991-11-01

For the quarantine treatment of rice-straw products from foreign countries, irradiation effects of gamma-rays and electron beams on plant pathogenic microorganisms especially on fungi were investigated. The total aerobic bacteria in rice-straw was determined to be 3x10{sup 7} - 3x10{sup 8} per gram which consisted mainly of Pseudomonas, Flavobacterium, Arthrobacter and Erwinia. The principal bacteria in rice-straw could be eliminated with 5 kGy of gamma irradiation. Deinococcus proteolyticus and Pseudomonas radiora were the main survivors at 5 to 12 kGy of irradiation. Saprophytic fungus which belongs to Dimorphospora also survived up to 8 kGy of irradiation. The D{sub 10} values of 26 strains of fungi isolated from rice-straw were 1.1 to 2.5 times higher in the dry condition compared to the values when irradiated in 0.067 M phosphate buffer solution. The induction dose in the dry condition also increased from 1.5 to 10 times than that in the wet condition. In the case of electron beam irradiation of fungi under dry conditions, D{sub 10} values were about 1.3 times higher than that of gamma irradiation. From this study, the dose necessary to reduce the plant pathogenic fungi in rice-straw at a level below 10{sup -4} per gram was estimated to be as 7-8 kGy for gamma-irradiation and 10 kGy for electron beam irradiation. (author).

Denitrification in groundwater at uranium mill tailings sites

Energy Technology Data Exchange (ETDEWEB)

Goering, Timothy J [Jacobs Engineering Group, Inc., Albuquerque, NM (United States); Groffman, Armando [Roy F. Weston, Inc., Albuquerque, NM (United States); Thomson, Bruce [University of New Mexico, Albuquerque, NM (United States)

1992-07-01

Nitrates are a major contaminant in groundwater at many Uranium Mill Tailings Remedial Action (UMTRA) sites. Microbial denitrification, the transformation of nitrate to nitrogen gas, may be occurring in groundwater at several UMTRA sites. Denitrification is a biologically mediated process whereby facultative anaerobes use nitrate for respiration under anaerobic conditions. Denitrifying bacteria are ubiquitous in soils, sediments, and water. Denitrification requires nitrate, organic carbon, oxygen-limiting conditions, and trace nutrients, especially phosphorus. The lack of organic carbon is the most common limiting factor for denitrification. Denitrification occurs under a limited range of temperature and pH. The uranium milling processes used at UMTRA sites provided a readily available source of carbon and nitrates for denitrifying bacteria. At the Maybell, Colorado, site, the denitrifying organisms Pseudomonas, Flavobacterium and Acinetobacter were identified in core samples of materials from beneath the tailings. In addition, microcosm experiments simulating aquifer conditions beneath the tailings pile showed an average 40 percent decrease in nitrate concentrations over 13 days. At the New Rifle, Colorado, site, aquifer conditions appear favorable for denitrification. Nitrate and organic carbon are readily available in the groundwater, and redox conditions beneath and downgradient of the tailings pile are relatively anoxic. Downgradient from the tailings, total nitrogen is being removed from the groundwater system at a greater rate than the geochemically conservative anion, chloride. This removal may be due to denitrification and adsorption of ammonium onto clay and silt particles. (author)

Diversity and Geographical Distribution of Flavobacterium psychrophilum Isolates and Their Phages: Patterns of Susceptibility to Phage Infection and Phage Host Range

DEFF Research Database (Denmark)

Castillo, Daniel; Christiansen, Rói Hammershaimb; Espejo, Romilio

2014-01-01

in disease control requires detailed knowledge about the diversity and dynamics of host susceptibility to phage infection. For this reason, we examined the genetic diversity of 49 F. psychrophilum strains isolated in three different areas (Chile, Denmark, and USA) through direct genome restriction enzyme...... analysis (DGREA) and their susceptibility to 33 bacteriophages isolated in Chile and Denmark, thus covering large geographical (>12,000 km) and temporal (>60 years) scales of isolation. An additional 40 phage-resistant isolates obtained from culture experiments after exposure to specific phages were...... examined for changes in phage susceptibility against the 33 phages. The F. psychrophilum and phage populations isolated from Chile and Denmark clustered into geographically distinct groups with respect to DGREA profile and host range, respectively. However, cross infection between Chilean phage isolates...

CARACTERIZACIÓN FENOTIPICA Y MOLECULAR DE POBLACIONES BACTERIANAS AISLADAS DE UN SUELO CONTAMINADO CON DIESEL Y SOMETIDO A DOS TECNOLOGÍAS DE BIORREMEDIACIÓN

Directory of Open Access Journals (Sweden)

ARRIETA R. OLGA M.

2012-12-01

Full Text Available En este estudio se presenta la biorremediación como alternativa para la recuperación de ecosistemas contaminados. En este trabajo se realizó un derrame experimental de diesel sobre un suelo de pastoreo, el cual fue remediado haciendo uso de dos tecnologías de biorremediación: atenuación natural; conocida como la capacidad natural de los microorganismos nativos para degradar en un tiempo determinado cualquier compuesto xenobiótico, y bioestimulación; conocida como la adición al medio de nutrientes (fosforo y nitrógeno con el fin de estimular el metabolismo de los microorganismos nativos, logrando con esto acelerar el proceso de degradación. Los resultados de la prueba de respirometría indicaron que ambos tratamientos produjeron valores significativos de remoción del hidrocarburo pero el tratamiento de bioestimulación se destacó con el 98,17% de degradación. Se obtuvieron siete aislamientos bacterianos a partir de las siembras realizadas, según la caracterización molecular pertenecen a los géneros: Enterobacter, Bacillus, Arthrobacter, Sanguibacter, Staphylococcus y Flavobacterium. Todos los aislamientos fueron capaces de metabolizar el diesel como fuente de carbono y energía; por ello y teniendo en cuenta que para algunos de estos microorganismos su papel en biorremediación no ha sido estudiado extensivamente, se recomienda continuar con su evaluación para conocer cuál es realmente su potencial para la solución de problemas ambientales.

Pyrosequencing based assessment of bacterial diversity in Turkish Rhipicephalus annulatus and Dermacentor marginatus ticks (Acari: Ixodidae).

Science.gov (United States)

Tekin, Saban; Dowd, Scot E; Davinic, Marko; Bursali, Ahmet; Keskin, Adem

2017-03-01

Ticks continue to be a threat to human and animal health in Turkey, as they are considered important vectors of human and animal diseases. The objectives of this investigation are to characterize the microbial communities of two tick species, Rhipicephalus annulatus and Dermacenter marginatus, analyze patterns of co-occurrence among microbial taxa, identify and compare pathogens contributing human diseases, and determine whether avirulent symbionts could exclude human pathogens from tick communities. Furthermore, this study explores a microbiome of the R. annulatus and D. marginatus via the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) technique to describe their bacterial diversity. Pyrosequencing was performed on adult males and females isolated from humans from two high-risk Turkish provinces, Sivas and Amasya, during tick outbreaks in 2009. A total of 36,253 sequences were utilized for analyses of the 8 tick samples. Several pathogenic genera such as Francisella, Coxiella, Rickettsia, and Shigella were detected in the ticks tested. The most distinguishable difference between the two species of ticks was the lack of known human pathogen Rickettsia in R. annulatus and in samples 9 and 10 of D. marginatus. These samples had higher relative abundance of Flavobacterium sp., Curvibacter sp., Acidovorax sp., and Bacteroidaceae genera mostly representing symbionts which form a large component of normal tick microbiota. The outcome of this study is consistent with the predictions of the community ecological theory that diversity-rich bacteriomes are more resistant to bacterial invasion (and consequent pathogen dissemination) than diversity-deprived ones.

Investigation of Arsenotrophic Microbiome in Arsenic-Affected Bangladesh Groundwater.

Science.gov (United States)

Sultana, Munawar; Mou, Taslin Jahan; Sanyal, Santonu Kumar; Diba, Farzana; Mahmud, Zahid Hayat; Parvez, Anowar Khasru; Hossain, M Anwar

2017-09-01

Arsenotrophic bacteria contribute to the nutrient cycling in arsenic (As) affected groundwater. This study employed a culture-independent and -dependent investigation of arsenotrophic microbiomes in As affected groundwater samples collected from Madhabpur, Sonatengra, and Union Porishod in Singair Upazila, Manikganj, Bangladesh. Total As contents, detected by Atomic Absorption Spectrophotometry (AAS) of the samples, were 47 µg/L (Madhabpur, SNGW-1), 53 µg/L (Sonatengra, SNGW-2), and 12 µg/L (Union porishod, SNGW-3), whereas the control well (SNGW-4; depths >150 m) showed As content of 6 µg/L. Denaturing Gradient Gel Electrophoresis (DGGE) analysis of the amplified 16S rRNA gene from As-affected groundwater samples revealed the dominance of aerobic bacteria Pseudomonas within heterogeneous bacterial populations. DGGE of heterotrophic enrichments supplemented with arsenite [As (III)] for 4 weeks showed the dominance of Chryseobacterium, Flavobacterium, and Aquabacterium, whereas the dominant genera in that of autotrophic enrichments were Aeromonas, Acinetobacter, and Pseudomonas. Cultured bacteria retrieved from both autotrophic and heterotrophic enrichments were distinguished into nine genotypes belonging to Chryseobacterium, Acinetobacter, Escherichia, Pseudomonas, Stenotrophomonas, Janibacter, Staphylococcus, and Bacillus. They exhibited varying range of As(III) tolerance from 4 to 27 mM. As(III) transformation potential was confirmed within the isolates with oxidation rate as high as 0.143 mM/h for Pseudomonas sp. Sn 28. The arsenotrophic microbiome specifies their potential role in groundwater As-cycling and their genetic information provide the scientific basis for As-bioremediation. © 2017, National Ground Water Association.

Effect of pine bark and compost on the biological denitrification process of non-hazardous landfill leachate: Focus on the microbiology

International Nuclear Information System (INIS)

Trois, Cristina; Coulon, Frederic; Polge de Combret, Cecile; Martins, Jean M.F.; Oxarango, Laurent

2010-01-01

In an attempt to optimize the cost-efficiency of landfill leachate treatment by biological denitrification process, our study focused on finding low-cost alternatives to traditional expensive chemicals such as composted garden refuse and pine bark, which are both available in large amount in South African landfill sites. The overall objective was to assess the behaviour of the bacterial community in relation to each substrate while treating high strength landfill leachates. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests with immature compost and pine bark. High strength leachate was simulated using a solution of water and nitrate at a concentration of 500 mg l -1 . Results suggest that pine bark released large amounts of phenolic compounds and hydroxylated benzene rings, which both can delay the acclimatization time and inhibit the biological denitrification (only 30% efficiency). Furthermore, presence of potential pathogens like Enterobacter and Pantoea agglomerans prevents the applicability of the pine bark in full-scale operations. On the other hand, lightly composted garden refuse (CGR) offered an adequate substrate for the formation of a biofilm necessary to complete the denitrification process (total nitrate removal observed within 7 days). CGR further contributed to a rapid establishment of an active consortium of denitrifiers including Acinetobacter, Rhizobium, Thermomonas, Rheinheimera, Phaeospirillum and Flavobacterium. Clearly the original composition, nature, carbon to nitrogen ratio (C/N) and degree of maturity and stability of the substrates play a key role in the denitrification process, impacting directly on the development of the bacterial population and, therefore, on the long-term removal efficiency.

Microbial Community Profile and Water Quality in a Protected Area of the Caatinga Biome.

Science.gov (United States)

Lopes, Fabyano Alvares Cardoso; Catão, Elisa Caldeira Pires; Santana, Renata Henrique; Cabral, Anderson de Souza; Paranhos, Rodolfo; Rangel, Thiago Pessanha; de Rezende, Carlos Eduardo; Edwards, Robert A; Thompson, Cristiane C; Thompson, Fabiano L; Kruger, Ricardo Henrique

2016-01-01

The Caatinga is a semi-arid biome in northeast Brazil. The Paraguaçú River is located in the Caatinga biome, and part of its course is protected by the National Park of Chapada Diamantina (PNCD). In this study we evaluated the effect of PNCD protection on the water quality and microbial community diversity of this river by analyzing water samples obtained from points located inside and outside the PNCD in both wet and dry seasons. Results of water quality analysis showed higher levels of silicate, ammonia, particulate organic carbon, and nitrite in samples from the unprotected area compared with those from protected areas. Pyrosequencing of the 16S rRNA genes revealed that Burkholderiales was abundant in samples from all three sites during both seasons and was represented primarily by the genus Polynucleobacter and members of the Comamonadaceae family (e.g., genus Limnohabitans). During the dry season, the unprotected area showed a higher abundance of Flavobacterium sp. and Arthrobacter sp., which are frequently associated with the presence and/or degradation of arsenic and pesticide compounds. In addition, genes that appear to be related to agricultural impacts on the environment, as well as those involved in arsenic and cadmium resistance, copper homeostasis, and propanediol utilization, were detected in the unprotected areas by metagenomic sequencing. Although PNCD protection improves water quality, agricultural activities around the park may affect water quality within the park and may account for the presence of bacteria capable of pesticide degradation and assimilation, evidencing possible anthropogenic impacts on the Caatinga.

Ulcerative enteritis in Homarus americanus: case report and molecular characterization of intestinal aerobic bacteria of apparently healthy lobsters in live storage.

Science.gov (United States)

Battison, Andrea L; Després, Béatrice M; Greenwood, Spencer J

2008-10-01

An intermoult male American lobster, Homarus americanus, with severe intestinal lesions was encountered while collecting samples of aerobic intestinal bacteria from lobsters held in an artificial sea-water recirculation aquarium system. Grossly, the intestine was firm, thickened, and white. Histologic examination revealed a severe, diffuse, ulcerative enteritis which spared the chitin-lined colon, somewhat similar to hemocytic enteritis of shrimp. The bacterial isolates from this lobster were compared to 11 other lobsters lacking gross intestinal lesions. Two organisms, one identified as Vibrio sp. and another most similar to an uncultured proteobacterium (98.9%), clustering with Rhanella and Serratia species using 16S rDNA PCR, were isolated from the intestines of the 11, grossly normal, lobsters and the affected lobster. An additional two intestinal isolates were cultured only from the lobster with ulcerative enteritis. One, a Flavobacterium, similar to Lutibacter litoralis (99.3%), possibly represented a previously described commensal of the distal intestine. The second, a Vibrio sp., was unique to the affected animal. While the etiology of the ulcerative enteritis remains undetermined, this report represents the first description of gross and histologic findings in H. americanus of a condition which has morphologic similarities to hemocytic enteritis of shrimp. An additional observation was a decrease in the number of intestinal isolates recovered from the 11 apparently healthy lobsters compared to that previously reported for recently harvested lobster. More comprehensive studies of the relationship between the health of lobsters, gut microbial flora and the husbandry and environment maintained within holding units are warranted.

Nitrogen cycling processes and microbial community composition in bed sediments in the Yukon River at Pilot Station

Science.gov (United States)

Repert, Deborah A.; Underwood, Jennifer C.; Smith, Richard L.; Song, Bongkeun

2014-01-01

Information on the contribution of nitrogen (N)-cycling processes in bed sediments to river nutrient fluxes in large northern latitude river systems is limited. This study examined the relationship between N-cycling processes in bed sediments and N speciation and loading in the Yukon River near its mouth at the Bering Sea. We conducted laboratory bioassays to measure N-cycling processes in sediment samples collected over distinct water cycle seasons. In conjunction, the microbial community composition in the bed sediments using genes involved in N-cycling (narG, napA, nosZ, and amoA) and 16S rRNA gene pyrosequences was examined. Temporal variation was observed in net N mineralization, nitrate uptake, and denitrification rate potentials and correlated strongly with sediment carbon (C) and extractable N content and microbial community composition rather than with river water nutrient concentrations. The C content of the bed sediment was notably impacted by the spring flood, ranging from 1.1% in the midst of an ice-jam to 0.1% immediately after ice-out, suggesting a buildup of organic material (OM) prior to scouring of the bed sediments during ice break up. The dominant members of the microbial community that explained differences in N-processing rates belonged to the genera Crenothrix,Flavobacterium, and the family of Comamonadaceae. Our results suggest that biogeochemical processing rates in the bed sediments appear to be more coupled to hydrology, nutrient availability in the sediments, and microbial community composition rather than river nutrient concentrations at Pilot Station.

Extraordinary proliferation of microorganisms in aposymbiotic pea aphids, Acyrthosiphon pisum.

Science.gov (United States)

Nakabachi, Atsushi; Ishikawa, Hajime; Kudo, Toshiaki

2003-03-01

Aposymbiotic pea aphids, which were deprived of their intracellular symbiotic bacterium, Buchnera, exhibit growth retardation and no fecundity. High performance liquid chromatographic (HPLC) analysis revealed that these aposymbiotic aphids, when reared on broad bean plants, accumulated a large amount of histamine. To assess the possibility of extraordinary proliferation of microorganisms other than Buchnera, we enumerated eubacteria and fungi in aphids using the real-time quantitative PCR method that targets genes encoding small-subunit rRNAs. The result showed that these microorganisms were extremely abundant in the aposymbiotic aphids reared on plants. Microbial communities in aposymbiotic aphids were further profiled by phylogenetic analysis of small-subunit rDNAs. Of 172 nonchimeric sequences of fungal 18S rDNAs, 138 (80.2%) belonged to the phylum Ascomycota. Among them, 21 clustered within a monophyletic group consisting of insect-pathogenic fungi and yeast-like symbionts of homopteran insects. Thirty-one (18.0%), two (1.2%), and one (0.6%) clones were clustered within the Basidiomycota, Zygomycota, and Oomycota, respectively. Of 167 nonchimeric sequences of eubacterial 16S rDNAs, 84 (50.3%) belonged to the gamma-subdivision of Proteobacteria to which most primary endosymbionts of insects and prolific histamine producers belong. Forty (24.0%), 25 (15.0%), 10 (6.0%), and five (3.0%) clones were clustered within alpha-Proteobacteria, Cytophaga-Flavobacterium-Bacteroides (CFB) group, Actinobacteria, and beta-Proteobacteria, respectively. Three had no phylogenetic association with known taxonomic divisions. None of the sequences studied in this study coincided exactly with those deposited in GenBank.

Bacterial community structure and diversity responses to the direct revegetation of an artisanal zinc smelting slag after 5 years.

Science.gov (United States)

Luo, Youfa; Wu, Yonggui; Wang, Hu; Xing, Rongrong; Zheng, Zhilin; Qiu, Jing; Yang, Lian

2018-05-01

This comparative field study examined the responses of bacterial community structure and diversity to the revegetation of zinc (Zn) smelting waste slag with eight plant species after 5 years. The microbial community structure of waste slag with and without vegetation was evaluated using high-throughput sequencing. The physiochemical properties of Zn smelting slag after revegetation with eight plant rhizospheres for 5 years were improved compared to those of bulk slag. Revegetation significantly increased the microbial community diversity in plant rhizospheres, and at the phylum level, Proteobacteria, Acidobacteria, and Bacteroidetes were notably more abundant in rhizosphere slags than those in bulk waste slag. Additionally, revegetation increased the relative abundance of plant growth-promoting rhizobacteria such as Flavobacterium, Streptomyces, and Arthrobacter as well as symbiotic N 2 fixers such as Bradyrhizobium. Three dominant native plant species (Arundo donax, Broussonetia papyrifera, and Robinia pseudoacacia) greatly increased the quality of the rhizosphere slags. Canonical correspondence analysis showed that the differences in bacterial community structure between the bulk and rhizosphere slags were explained by slag properties, i.e., pH, available copper (Cu) and lead (Pb), moisture, available nitrogen (N), phosphorus (P), and potassium (K), and organic matter (OM); however, available Zn and cadmium (Cd) contents were the slag parameters that best explained the differences between the rhizosphere communities of the eight plant species. The results suggested that revegetation plays an important role in enhancing bacterial community abundance and diversity in rhizosphere slags and that revegetation may also regulate microbiological properties and diversity mainly through changes in heavy metal bioavailability and physiochemical slag characteristics.

Radiation preservation of mackerel. Part of a coordinated programme on radiation preservation of Asian fish and fishery products

International Nuclear Information System (INIS)

Hussain, A.M.; Ashraf Chaudry, M.

1979-10-01

Effect of irradiation on shelf-life extension of mackerel (Rastrelliger kanagurta) stored at different temperatures was investigated. The microbial load in the unirradiated samples stored at both the temperatures reached the acceptability limit after 7 days at 1-3 0 C and after 3 days at 7-8 0 C. In the irradiated (150 krad) samples the same level of bacterial acceptability was attained between 2-3 weeks at 1-3 0 C and between 7-11 days at 7-8 0 C. In the unirradiated samples Pseudomonas and Proteus dominated after one month storage at 1-3 0 C whereas at 7-8 0 C, Bacillus was also encountered in addition to Pseudomonas and Proteus during the entire storage period. In the irradiated samples at 1-3 0 C, Achromobacter and Flavobacterium were dominant while Bacillus and Micrococcus were also prominent. At 7-8 0 C Achromobacter-Moraxella and Bacillus were dominant in all the irradiated samples, while at higher doses Sarcina and Lactobacillus were also isolated in high numbers. Moisture and protein contents decreased and driploss increased significantly during storage of fish. Dipping of fish fillets in 10% SPP for 10 minutes minimized the driploss significantly. A dose of 150 krad in combination with 10% SPP treatment could extend the market-life of mackerel for 2 weeks over unirradiated control and prevent loss of drip during storage at 1-3 0 C. At 7-8 0 C, one week extension in shelf-life was obtained on the basis of microbial studies

The use of phage FCL-2 as an alternative to chemotherapy against columnaris disease in aquaculture

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Elina eLaanto

2015-08-01

Full Text Available Flavobacterium columnare, the causative agent of columnaris disease in fish, causes millions of dollars of losses in the US channel catfish industry alone, not to mention aquaculture industry worldwide. Novel methods are needed for the control and treatment of bacterial diseases in aquaculture to replace traditionally used chemotherapies. A potential solution could be the use of phages, i.e., bacterial viruses, host-specific and self-enriching particles that can be can easily distributed via water flow. We examined the efficacy of phages to combat columnaris disease. A previously isolated phage, FCL-2, infecting F. columnare, was characterized by sequencing. The 47 142 bp genome of the phage had G + C content of 30.2%, and the closest similarities regarding the structural proteins were found in Cellulophaga phage phiSM. Under controlled experimental conditions, two host fish species, rainbow trout (Oncorhynchus mykiss and zebrafish (Danio rerio, were used to study the success of phage therapy to prevent F. columnare infections. The survival of both fish species was significantly higher in the presence of the phage. Hundred percent of the zebrafish and 50 % of the rainbow trout survived in the phage treatment (survival without phage 0 % and 8.3 %, respectively. Most importantly, the rainbow trout population was rescued from infection by a single addition of the phage into the water in a flow-through fish tank system. Thus, F. columnare could be used as a model system to test the benefits and risks of phage therapy on a larger scale.

Diversity of ionizing radiation-resistant bacteria obtained from the Taklimakan Desert.

Science.gov (United States)

Yu, Li Zhi-Han; Luo, Xue-Song; Liu, Ming; Huang, Qiaoyun

2015-01-01

So far, little is known about the diversity of the radiation-resistant microbes of the hyperarid Taklimakan Desert. In this study, ionizing radiation (IR)-resistant bacteria from two sites in Xinjiang were investigated. After exposing the arid (water content of 0.8 ± 0.3%) and non-arid (water content of 21.3 ± 0.9%) sediment samples to IR of 3000 Gy using a (60)Co source, a total of 52 γ-radiation-resistant bacteria were isolated from the desert sample. The 16S rRNA genes of all isolates were sequenced. The phylogenetic tree places these isolates into five groups: Cytophaga-Flavobacterium-Bacteroides, Proteobacteria, Deinococcus-Thermus, Firmicutes, and Actinobacteria. Interestingly, this is the first report of radiation-resistant bacteria belonging to the genera Knoellia, Lysobacter, Nocardioides, Paracoccus, Pontibacter, Rufibacter and Microvirga. The 16s rRNA genes of four isolates showed low sequence similarities to those of the published species. Phenotypic analysis showed that all bacteria in this study are able to produce catalase, suggesting that these bacteria possess reactive oxygen species (ROS)-scavenging enzymes. These radiation-resistant bacteria also displayed diverse metabolic properties. Moreover, their radiation resistances were found to differ. The diversity of the radiation-resistant bacteria in the desert provides further ecological support for the hypothesis that the ionizing-radiation resistance phenotype is a consequence of the evolution of ROS-scavenging systems that protect cells against oxidative damage caused by desiccation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effect of pine bark and compost on the biological denitrification process of non-hazardous landfill leachate: Focus on the microbiology

Energy Technology Data Exchange (ETDEWEB)

Trois, Cristina, E-mail: troisc@ukzn.ac.za [Centre for Research in Environmental, Coastal and Hydrological Engineering, School of Civil Engineering, Surveying and Construction, University of KwaZulu-Natal, Howard College Campus, Durban 4041 (South Africa); Coulon, Frederic; Polge de Combret, Cecile [Centre for Resource Management and Efficiency, School of Applied Sciences, Cranfield University, MK43 0AL (United Kingdom); Martins, Jean M.F.; Oxarango, Laurent [Laboratoire d' etude de Transferts en Hydrologie et Environnement, UMR 5564 (CNRS/INPG/IRD/UJF), Universite de Grenoble, BP 53, 38041 Grenoble Cedex 9 (France)

2010-09-15

In an attempt to optimize the cost-efficiency of landfill leachate treatment by biological denitrification process, our study focused on finding low-cost alternatives to traditional expensive chemicals such as composted garden refuse and pine bark, which are both available in large amount in South African landfill sites. The overall objective was to assess the behaviour of the bacterial community in relation to each substrate while treating high strength landfill leachates. Denitrification processes in fixed bed reactors were simulated at laboratory scale using anaerobic batch tests with immature compost and pine bark. High strength leachate was simulated using a solution of water and nitrate at a concentration of 500 mg l{sup -1}. Results suggest that pine bark released large amounts of phenolic compounds and hydroxylated benzene rings, which both can delay the acclimatization time and inhibit the biological denitrification (only 30% efficiency). Furthermore, presence of potential pathogens like Enterobacter and Pantoea agglomerans prevents the applicability of the pine bark in full-scale operations. On the other hand, lightly composted garden refuse (CGR) offered an adequate substrate for the formation of a biofilm necessary to complete the denitrification process (total nitrate removal observed within 7 days). CGR further contributed to a rapid establishment of an active consortium of denitrifiers including Acinetobacter, Rhizobium, Thermomonas, Rheinheimera, Phaeospirillum and Flavobacterium. Clearly the original composition, nature, carbon to nitrogen ratio (C/N) and degree of maturity and stability of the substrates play a key role in the denitrification process, impacting directly on the development of the bacterial population and, therefore, on the long-term removal efficiency.

Characterization of multiple antibiotic resistance of culturable microorganisms and metagenomic analysis of total microbial diversity of marine fish sold in retail shops in Mumbai, India.

Science.gov (United States)

Naik, Onkar A; Shashidhar, Ravindranath; Rath, Devashish; Bandekar, Jayant R; Rath, Archana

2018-03-01

Marine fish species were analyzed for culturable and total metagenomic microbial diversity, antibiotic resistance (AR) pattern, and horizontal gene transfer in culturable microorganisms. We observed a high AR microbial load of 3 to 4 log CFU g -1 . Many fish pathogens like Providencia, Staphylococcus, Klebsiella pneumoniae, Enterobacter, Vagococcus, and Aeromonas veronii were isolated. Photobacterium and Vibrio were two major fish and human pathogens which were identified in the fish metagenome. Other pathogens that were identified were Shewanella, Acinetobacter, Psychrobacter, and Flavobacterium. Most of these pathogens were resistant to multiple antibiotics such as erythromycin, kanamycin, neomycin, streptomycin, penicillin, cefotaxime, bacitracin, rifampicin, trimethoprim, ciprofloxacin, and doxycycline with a high multiple antibiotic resistance index of 0.54-0.77. The fish microflora showed high prevalence of AR genes like bla TEM , Class I integron, tetA, aph(3')-IIIa, ermB, aadA, and sul1. Nineteen of 26 AR isolates harbored Class I integrons showing high co-resistance to trimethoprim, kanamycin, doxycycline, and cefotaxime. Mobile R-plasmids from 6 of the 12 AR pathogens were transferred to recipient E. coli after conjugation. The transconjugants harbored the same R-plasmid carrying bla CTX-M , dfr1, tetA, bla TEM , and cat genes. This study confirms that fish is a potential carrier of AR pathogens which can enter the human gut via food chain. To the best of our knowledge, this is the first study in the Indian subcontinent reporting a direct evidence of spread of AR pathogens to humans from specific marine fish consumption.

Biochar in co-contaminated soil manipulates arsenic solubility and microbiological community structure, and promotes organochlorine degradation.

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Samuel J Gregory

Full Text Available We examined the effect of biochar on the water-soluble arsenic (As concentration and the extent of organochlorine degradation in a co-contaminated historic sheep-dip soil during a 180-d glasshouse incubation experiment. Soil microbial activity, bacterial community and structure diversity were also investigated. Biochar made from willow feedstock (Salix sp was pyrolysed at 350 or 550°C and added to soil at rates of 10 g kg-1 and 20 g kg-1 (representing 30 t ha-1 and 60 t ha-1. The isomers of hexachlorocyclohexane (HCH alpha-HCH and gamma-HCH (lindane, underwent 10-fold and 4-fold reductions in concentration as a function of biochar treatment. Biochar also resulted in a significant reduction in soil DDT levels (P < 0.01, and increased the DDE:DDT ratio. Soil microbial activity was significantly increased (P < 0.01 under all biochar treatments after 60 days of treatment compared to the control. 16S amplicon sequencing revealed that biochar-amended soil contained more members of the Chryseobacterium, Flavobacterium, Dyadobacter and Pseudomonadaceae which are known bioremediators of hydrocarbons. We hypothesise that a recorded short-term reduction in the soluble As concentration due to biochar amendment allowed native soil microbial communities to overcome As-related stress. We propose that increased microbiological activity (dehydrogenase activity due to biochar amendment was responsible for enhanced degradation of organochlorines in the soil. Biochar therefore partially overcame the co-contaminant effect of As, allowing for enhanced natural attenuation of organochlorines in soil.

The Influence of Infective Dose on the Virulence of a Generalist Pathogen in Rainbow Trout (Oncorhynchus mykiss and Zebra Fish (Danio rerio.

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Hanna Kinnula

Full Text Available Pathogen density and genetic diversity fluctuate in the outside-host environment during and between epidemics, affecting disease emergence and the severity and probability of infections. Although the importance of these factors for pathogen virulence and infection probability has been acknowledged, their interactive effects are not well understood. We studied how an infective dose in an environmentally transmitted opportunistic fish pathogen, Flavobacterium columnare, affects its virulence both in rainbow trout, which are frequently infected at fish farms, and in zebra fish, a host that is not naturally infected by F. columnare. We used previously isolated strains of confirmed high and low virulence in a single infection and in a co-infection. Infection success (measured as host morbidity correlated positively with dose when the hosts were exposed to the high-virulence strain, but no response for the dose increase was found when the hosts were exposed to the low-virulence strain. Interestingly, the co-infection resulted in poorer infection success than the single infection with the high-virulence strain. The rainbow trout were more susceptible to the infection than the zebra fish but, in both species, the effects of the doses and the strains were qualitatively similar. We suggest that as an increase in dose can lead to increased host morbidity, both the interstrain interactions and differences in infectivity in different hosts may influence the severity and consequently the evolution of disease. Our results also confirm that the zebra fish is a good laboratory model to study F. columnare infection.

Effective pesticide nano formulations and their bacterial degradation

Science.gov (United States)

Ramadass, M.; Thiagarajan, P.

2017-11-01

The use of chemical pesticides for agricultural pest control and the consequent damage to the ecosystem at air, water and soil levels has become a factor of common knowledge. This alarming trend has led to research and development in the area of nanoformulations to achieve the end use of pest control with very low concentrations of pesticides. Such formulations are being proven to be as effective as traditional formulations due to their inherent ability to achieve controlled delivery of their respective active ingredients. The end result is a successful pest control with minimum environmental damage. Despite this, certain organic groups, that form the essential structural constituents of these pesticides, are not readily degraded due to their complex nature. They continue to persist, accumulate and biomagnify in the environment leading to short and long term hazards. In this context, it has been noted that certain common genera of bacteria such as Bacillus, Pseudomonas, Flavobacterium, Sphingomonas, Brevibacterium, Burkholderia, etc possess the inherent ability to utilise specific chemical groups in the pesticides as their sole source of either carbon and / or nitrogen and consequently achieve their conversion into non-toxic end products. A potential bioremediation process is thus slowly gaining popularity and being implemented on a pilot scale. However, large scale successful pesticide microbial remediation will involve experimentation with several combinations of a variety of nano pesticide formulations with different genera of bacteria under optimised conditions. Such studies will throw light on the precise genus and species of bacteria that may degrade the required groups of pesticides, for environmental damage control in the long run.

Mortality of centrarchid fishes in the Potomac drainage: survey results and overview of potential contributing factors.

Science.gov (United States)

Blazer, V S; Iwanowicz, L R; Starliper, C E; Iwanowicz, D D; Barbash, P; Hedrick, J D; Reeser, S J; Mullican, J E; Zaugg, S D; Burkhardt, M R; Kelble, J

2010-09-01

Skin lesions and spring mortality events of smallmouth bass Micropterus dolomieu and selected other species were first noted in the South Branch of the Potomac River in 2002. Since that year morbidity and mortality have also been observed in the Shenandoah and Monocacy rivers. Despite much research, no single pathogen, parasite, or chemical cause for the lesions and mortality has been identified. Numerous parasites, most commonly trematode metacercariae and myxozoans; the bacterial pathogens Aeromonas hydrophila, Aeromonas salmonicida, and Flavobacterium columnare; and largemouth bass virus have all been observed. None have been consistently isolated or observed at all sites, however, nor has any consistent microscopic pathology of the lesions been observed. A variety of histological changes associated with exposure to environmental contaminants or stressors, including intersex (testicular oocytes), high numbers of macrophage aggregates, oxidative damage, gill lesions, and epidermal papillomas, were observed. The findings indicate that selected sensitive species may be stressed by multiple factors and constantly close to the threshold between a sustainable (healthy) and nonsustainable (unhealthy) condition. Fish health is often used as an indicator of aquatic ecosystem health, and these findings raise concerns about environmental degradation within the Potomac River drainage. Unfortunately, while much information has been gained from the studies conducted to date, due to the multiple state jurisdictions involved, competing interests, and other issues, there has been no coordinated approach to identifying and mitigating the stressors. This synthesis emphasizes the need for multiyear, interdisciplinary, integrative research to identify the underlying stressors and possible management actions to enhance ecosystem health.

Aquiflexum balticum gen. nov., sp. nov., a novel marine bacterium of the Cytophaga-Flavobacterium-Bacteroides group isolated from surface water of the central Baltic Sea.

Science.gov (United States)

Brettar, Ingrid; Christen, Richard; Höfle, Manfred G

2004-11-01

A bacterial isolate from the Baltic Sea, BA160(T), was characterized for its physiological and biochemical features, fatty acid profile, G+C content and phylogenetic position based on 16S rRNA gene sequences. The strain was isolated from the surface water of the central Baltic Sea during the decay of a plankton bloom. Phylogenetic analyses of the 16S rRNA gene sequence revealed a clear affiliation with the family 'Flexibacteraceae', and showed the closest phylogenetic relationship with the species Belliella baltica and Cyclobacterium marinum. The G+C content of the DNA was 38.4 mol%. The strain was red-coloured due to carotenoids, Gram-negative, rod-shaped, and catalase- and oxidase-positive. Growth was observed at salinities from 0 to 6 %, with an optimum around 1.5 %. Temperature for growth ranged from 4 to 40 degrees C, with an optimum around 30 degrees C. The fatty acids were dominated by branched-chain fatty acids (>87 %), with a high abundance of iso-C(15 : 0) (23 %) and anteiso-C(15 : 0) (19 %). According to its morphology, physiology, fatty acid composition, G+C content and 16S rRNA gene sequence, strain BA160(T) is considered to represent a new genus of the family 'Flexibacteraceae'. Due to its aquatic origin, the name Aquiflexum balticum gen. nov, sp. nov. is suggested for the type species (type strain, BA160(T)=DSM 16537(T)=LMG 22565(T)=CIP 108445(T)) of the new genus.

Ação antagônica de rizobactérias contra Phytophthora parasitica e p. citrophthora e seu efeito no desenvolvimento de plântulas de citros

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AMORIM EDNA PEIXOTO DA ROCHA

2002-01-01

Full Text Available O antagonismo de Pseudomonas putida biovar A (C1-1B, P. putida biovar B (Santa Bárbara, P. fluorescens (C2-8C e RA2, Bacillus subtilis (OG e RC2 e Flavobacterium sp. (CIS/NA contra Phytophthora parasitica e P. citrophthora , agentes da podridão radicular dos citros, foi avaliado através da inibição do crescimento micelial (cultura pareada e redução na percentagem de infecção da doença em mudas de citros (tratamento de sementes com rizobactérias. Na seleção preliminar, 33 isolados bacterianos foram testados. Sementes de citros pré-germinadas foram tratadas por imersão nas suspensões das bactérias (10(9 ufc/ml, e plantadas em tubetes contendo solo natural infestado com o fitopatógeno (50 ml de suspensão/ kg de solo. A avaliação da percentagem de infecção foi efetuada após 15 dias. In vitro, os isolados bacterianos RC2, OG, CIS/NA e C1-1B foram os mais ativos inibidores do crescimento micelial de Phytophthora. Em condições de casa de vegetação, todos os isolados proporcionaram redução na percentagem de infecção da doença em todos os ensaios realizados. Promoção de crescimento de plantas foi verificada pela inoculação de plântulas com as linhagens OG, RC2, CiS/Na e C1-1B.

Basin-scale seasonal changes in marine free-living bacterioplankton community in the Ofunato Bay

KAUST Repository

Reza, Md. Shaheed

2018-04-26

The Ofunato Bay in the northeastern Pacific Ocean area of Japan possesses the highest biodiversity of marine organisms in the world and has attracted much attention due to its economic and environmental importance. We report here a shotgun metagenomic analysis of the year-round variation in free-living bacterioplankton collected across the entire length of the bay. Phylogenetic differences among spring, summer, autumn and winter bacterioplankton suggested that members of Proteobacteria tended to decrease at high water temperatures and increase at low temperatures. It was revealed that Candidatus Pelagibacter varied seasonally, reaching as much as 60% of all sequences at the genus level in the surface waters during winter. This increase was more evident in the deeper waters, where they reached up to 75%. The relative abundance of Planktomarina also rose during winter and fell during summer. A significant component of the winter bacterioplankton community was Archaea (mainly represented by Nitrosopumilus), as their relative abundance was very low during spring and summer but high during winter. In contrast, Actinobacteria and Cyanobacteria appeared to be higher in abundance during high-temperature periods. It was also revealed that Bacteroidetes constituted a significant component of the summer bacterioplankton community, being the second largest bacterial phylum detected in the Ofunato Bay. Its members, notably Polaribacter and Flavobacterium, were found to be high in abundance during spring and summer, particularly in the surface waters. Principal component analysis and hierarchal clustering analyses showed that the bacterial communities in the Ofunato Bay changed seasonally, likely caused by the levels of organic matter, which would be deeply mixed with surface runoff in the winter.

Evaluation of the organophosphorus hydrolase enzyme activity in creams and investigation of its stability

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Mariye Rajaie

2016-06-01

Full Text Available The main purpose of this project is investigation of the organophosphorus hydrolase (OPH enzyme activity in water in oil (w/o and oil in water (o/w creams and investigation of the OPH enzyme stability in formulated creams. OPH enzyme was extracted and purified from strain flavobacterium. The w/o and o/w creams were prepared using different formulations. In order to achieve an emulsion with maximum stability, appropriate percentage of the cream components was selected by studying different formulations and the physical and chemical stability of the produced cream were considered. 5Uenzyme/90gcream enzyme was used for each formulation. To measure the enzyme activity in creams, extraction method was used and enzyme activity was determined based on parathion hydrolysis. The thermal stability of OPH in both types of w/o and o/w creams was studied at 4 and 30  °C for various time periods. The average enzyme activity was about 0.0065 U/gcream and 0.018 U/gcream for w/o and o/w creams respectivly. According to the results, the relative activity at 4 °C was reduced to 50% after 26 and 45 days in w/o and o/w creams, respectivly. The results showed that the OPH enzyme activity in o/w cream was 2.6 times more than that of w/o cream, because of the higher hydrophobicity of o/w cream compared to w/o. The OPH enzyme stability in o/w cream was greater in comparison to w/o cream. The OPH enzyme was active for nearly 2 months on o/w creams at 4 °C .

Microbial pollution indicators and culturable heterotrophic bacteria in a Mediterranean area (Southern Adriatic Sea Italian coasts)

Science.gov (United States)

Stabili, L.; Cavallo, R. A.

2011-05-01

In the present study we evaluated the degree of microbial water pollution along the coast line between Brindisi and Santa Maria di Leuca (Southern Adriatic Sea) as well as the culturable heterotrophic bacteria abundances and biodiversity in relation to the microbiological quality of the water. A total of 3773 colonies were isolated, subcultured and identified by several morphological, cultural and biochemical methods including the standardized API 20 E and API 20 NE tests. Along the examined coastal tract the microbial pollution indicators were always below the tolerance limits for bathing waters defined by the CEE directive, suggesting a good sanitary quality. Concerning culturable heterotrophic bacteria, different temporal density trends were observed in the four sites in relation to their geographical position. A positive relationship between the bacterial abundances and the temperature was observed in S. Cataldo and Otranto. The culturable bacterial community was mainly composed of the genera Aeromonas, Pseudomonas, Photobacterium and Flavobacterium. The Enterobacteriaceae family represented a conspicuous component of the bacterial community too. Bacilli were predominant among the Gram-positive bacteria. Of interest is the isolation of yeasts (2% at the surface and 1% at the bottom) taking into account their capability of biodegradation of various materials. Because of the low level of microbial pollution recorded, our results are indicative of the natural variation and diversity of the culturable bacterial community in such an oligotrophic ecosystem and could represent a good point of comparison with other ecosystems as well as a baseline for long term studies aimed to evaluate the effects of environmental fluctuations and human impacts on this aspect of biodiversity in coastal areas.

Microbiota bacteriana de ovos de jacaré-do-pantanal incubados naturalmente

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R. Pulcherio

Full Text Available RESUMO A cadeia produtiva do Caiman yacare tem-se destacado no Mato Grosso com a exportação de 143.386 peles em 2015, cujo sistema de manejo (ranching implica a incubação artificial dos ovos. Nesse processo, a contaminação bacteriana de ovos influencia a taxa de eclosão. O conhecimento da microbiota de ovos incubados naturalmente orienta o manejo sanitário adequado no incubatório. No presente estudo, são apresentadas informações sobre essa microbiota e sua correlação com a de outros crocodilianos, apontando-se as espécies com potencial patogênico. Amostras de 20 ninhos de C. yacare foram coletadas e semeadas em ágar sangue e ágar Mac Conckey. A colônia condizente com Salmonella sp. foi confirmada pela técnica de reação em cadeia de polimerase. Das 22 espécies bacterianas isoladas, 59% pertencem à família Enterobacteriaceae e 41% a outros táxons bacterianos. A semelhança dos achados com as bactérias isoladas na microbiota oral e/ou intestinal/cloacal de crocodilianos foi de 77,27%. As bactérias mais e menos frequentes foram, respectivamente, Bacillus cereus, Flavobacterium multivorum, Citrobacter freundii, Escherichia hermannii, Hafnia alvei, Morganella, morganni, Salmonella sp., Serratia marcescens e Shigella sonnei. Das bactérias isoladas, 86,36% têm potencial patogênico para crocodilianos. A origem materna e a ambiental da microbiota de ovos incubados naturalmente são, respectivamente, de 77,27% e 27,27%.

Epilithic Cyanobacterial Communities of a Marine Tropical Beach Rock (Heron Island, Great Barrier Reef): Diversity and Diazotrophy▿

Science.gov (United States)

Díez, Beatriz; Bauer, Karolina; Bergman, Birgitta

2007-01-01

The diversity and nitrogenase activity of epilithic marine microbes in a Holocene beach rock (Heron Island, Great Barrier Reef, Australia) with a proposed biological calcification “microbialite” origin were examined. Partial 16S rRNA gene sequences from the dominant mat (a coherent and layered pink-pigmented community spread over the beach rock) and biofilms (nonstratified, differently pigmented microbial communities of small shallow depressions) were retrieved using denaturing gradient gel electrophoresis (DGGE), and a clone library was retrieved from the dominant mat. The 16S rRNA gene sequences and morphological analyses revealed heterogeneity in the cyanobacterial distribution patterns. The nonheterocystous filamentous genus Blennothrix sp., phylogenetically related to Lyngbya, dominated the mat together with unidentified nonheterocystous filaments of members of the Pseudanabaenaceae and the unicellular genus Chroococcidiopsis. The dominance and three-dimensional intertwined distribution of these organisms were confirmed by nonintrusive scanning microscopy. In contrast, the less pronounced biofilms were dominated by the heterocystous cyanobacterial genus Calothrix, two unicellular Entophysalis morphotypes, Lyngbya spp., and members of the Pseudanabaenaceae family. Cytophaga-Flavobacterium-Bacteroides and Alphaproteobacteria phylotypes were also retrieved from the beach rock. The microbial diversity of the dominant mat was accompanied by high nocturnal nitrogenase activities (as determined by in situ acetylene reduction assays). A new DGGE nifH gene optimization approach for cyanobacterial nitrogen fixers showed that the sequences retrieved from the dominant mat were related to nonheterocystous uncultured cyanobacterial phylotypes, only distantly related to sequences of nitrogen-fixing cultured cyanobacteria. These data stress the occurrence and importance of nonheterocystous epilithic cyanobacteria, and it is hypothesized that such epilithic cyanobacteria

Evaluation of biostimulation and Tween 80 addition for the bioremediation of long-term DDT-contaminated soil.

Science.gov (United States)

Betancur-Corredor, Bibiana; Pino, Nancy J; Cardona, Santiago; Peñuela, Gustavo A

2015-02-01

The bioremediation of a long-term contaminated soil through biostimulation and surfactant addition was evaluated. The concentrations of 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) and its metabolites 1,1-dichloro-2,2-bis(4-chlorophenyl) ethane (DDD) and 1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene (DDE) were monitored during an 8-week remediation process. Physicochemical characterization of the treated soil was performed before and after the bioremediation process. The isolation and identification of predominant microorganisms during the remediation process were also carried out. The efficiency of detoxification was evaluated after each bioremediation protocol. Humidity and pH and the heterotrophic microorganism count were monitored weekly. The DDT concentration was reduced by 79% after 8 weeks via biostimulation with surfactant addition (B+S) and 94.3% via biostimulation alone (B). Likewise, the concentrations of the metabolites DDE and DDD were reduced to levels below the quantification limits. The microorganisms isolated during bioremediation were identified as Bacillus thuringiensis, Flavobacterium sp., Cuprivadius sp., Variovorax soli, Phenylobacterium sp. and Lysobacter sp., among others. Analysis with scanning electron microscopy (SEM) allowed visualization of the colonization patterns of soil particles. The toxicity of the soil before and after bioremediation was evaluated using Vibrio fischeri as a bioluminescent sensor. A decrease in the toxic potential of the soil was verified by the increase of the concentration/effect relationship EC50 to 26.9% and 27.2% for B+S and B, respectively, compared to 0.4% obtained for the soil before treatment and 2.5% by natural attenuation after 8 weeks of treatment. Copyright © 2014. Published by Elsevier B.V.

Low temperature S(0) biomineralization at a supraglacial spring system in the Canadian High Arctic.

Science.gov (United States)

Gleeson, D F; Williamson, C; Grasby, S E; Pappalardo, R T; Spear, J R; Templeton, A S

2011-07-01

Elemental sulfur (S(0) ) is deposited each summer onto surface ice at Borup Fiord pass on Ellesmere Island, Canada, when high concentrations of aqueous H(2) S are discharged from a supraglacial spring system. 16S rRNA gene clone libraries generated from sulfur deposits were dominated by β-Proteobacteria, particularly Ralstonia sp. Sulfur-cycling micro-organisms such as Thiomicrospira sp., and ε-Proteobacteria such as Sulfuricurvales and Sulfurovumales spp. were also abundant. Concurrent cultivation experiments isolated psychrophilic, sulfide-oxidizing consortia, which produce S(0) in opposing gradients of Na(2) S and oxygen. 16S rRNA gene analyses of sulfur precipitated in gradient tubes show stable sulfur-biomineralizing consortia dominated by Marinobacter sp. in association with Shewanella, Loktanella, Rubrobacter, Flavobacterium, and Sphingomonas spp. Organisms closely related to cultivars appear in environmental 16S rRNA clone libraries; none currently known to oxidize sulfide. Once consortia were simplified to Marinobacter and Flavobacteria spp. through dilution-to-extinction and agar removal, sulfur biomineralization continued. Shewanella, Loktanella, Sphingomonas, and Devosia spp. were also isolated on heterotrophic media, but none produced S(0) alone when reintroduced to Na(2) S gradient tubes. Tubes inoculated with a Marinobacter and Shewanella spp. co-culture did show sulfur biomineralization, suggesting that Marinobacter may be the key sulfide oxidizer in laboratory experiments. Light, florescence and scanning electron microscopy of mineral aggregates produced in Marinobacter experiments revealed abundant cells, with filaments and sheaths variably mineralized with extracellular submicron sulfur grains; similar biomineralization was not observed in abiotic controls. Detailed characterization of mineral products associated with low temperature microbial sulfur-cycling may provide biosignatures relevant to future exploration of Europa and Mars. © 2011

Aura-biomes are present in the water layer above coral reef benthic macro-organisms

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Kevin Walsh

2017-08-01

Full Text Available As coral reef habitats decline worldwide, some reefs are transitioning from coral- to algal-dominated benthos with the exact cause for this shift remaining elusive. Increases in the abundance of microbes in the water column has been correlated with an increase in coral disease and reduction in coral cover. Here we investigated how multiple reef organisms influence microbial communities in the surrounding water column. Our study consisted of a field assessment of microbial communities above replicate patches dominated by a single macro-organism. Metagenomes were constructed from 20 L of water above distinct macro-organisms, including (1 the coral Mussismilia braziliensis, (2 fleshy macroalgae (Stypopodium, Dictota and Canistrocarpus, (3 turf algae, and (4 the zoanthid Palythoa caribaeorum and were compared to the water microbes collected 3 m above the reef. Microbial genera and functional potential were annotated using MG-RAST and showed that the dominant benthic macro-organisms influence the taxa and functions of microbes in the water column surrounding them, developing a specific “aura-biome”. The coral aura-biome reflected the open water column, and was associated with Synechococcus and functions suggesting oligotrophic growth, while the fleshy macroalgae aura-biome was associated with Ruegeria, Pseudomonas, and microbial functions suggesting low oxygen conditions. The turf algae aura-biome was associated with Vibrio, Flavobacterium, and functions suggesting pathogenic activity, while zoanthids were associated with Alteromonas and functions suggesting a stressful environment. Because each benthic organism has a distinct aura-biome, a change in benthic cover will change the microbial community of the water, which may lead to either the stimulation or suppression of the recruitment of benthic organisms.

Aura-biomes are present in the water layer above coral reef benthic macro-organisms.

Science.gov (United States)

Walsh, Kevin; Haggerty, J Matthew; Doane, Michael P; Hansen, John J; Morris, Megan M; Moreira, Ana Paula B; de Oliveira, Louisi; Leomil, Luciana; Garcia, Gizele D; Thompson, Fabiano; Dinsdale, Elizabeth A

2017-01-01

As coral reef habitats decline worldwide, some reefs are transitioning from coral- to algal-dominated benthos with the exact cause for this shift remaining elusive. Increases in the abundance of microbes in the water column has been correlated with an increase in coral disease and reduction in coral cover. Here we investigated how multiple reef organisms influence microbial communities in the surrounding water column. Our study consisted of a field assessment of microbial communities above replicate patches dominated by a single macro-organism. Metagenomes were constructed from 20 L of water above distinct macro-organisms, including (1) the coral Mussismilia braziliensis , (2) fleshy macroalgae ( Stypopodium , Dictota and Canistrocarpus ), (3) turf algae, and (4) the zoanthid Palythoa caribaeorum and were compared to the water microbes collected 3 m above the reef. Microbial genera and functional potential were annotated using MG-RAST and showed that the dominant benthic macro-organisms influence the taxa and functions of microbes in the water column surrounding them, developing a specific "aura-biome". The coral aura-biome reflected the open water column, and was associated with Synechococcus and functions suggesting oligotrophic growth, while the fleshy macroalgae aura-biome was associated with Ruegeria , Pseudomonas, and microbial functions suggesting low oxygen conditions. The turf algae aura-biome was associated with Vibrio, Flavobacterium, and functions suggesting pathogenic activity, while zoanthids were associated with Alteromonas and functions suggesting a stressful environment. Because each benthic organism has a distinct aura-biome, a change in benthic cover will change the microbial community of the water, which may lead to either the stimulation or suppression of the recruitment of benthic organisms.

Prevalent bacterial species and novel phylotypes in advanced noma lesions.

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Paster, B J; Falkler Jr, W A; Enwonwu, C O; Idigbe, E O; Savage, K O; Levanos, V A; Tamer, M A; Ericson, R L; Lau, C N; Dewhirst, F E

2002-06-01

The purpose of this study was to determine the bacterial diversity in advanced noma lesions using culture-independent molecular methods. 16S ribosomal DNA bacterial genes from DNA isolated from advanced noma lesions of four Nigerian children were PCR amplified with universally conserved primers and spirochetal selective primers and cloned into Escherichia coli. Partial 16S rRNA sequences of approximately 500 bases from 212 cloned inserts were used initially to determine species identity or closest relatives by comparison with sequences of known species or phylotypes. Nearly complete sequences of approximately 1,500 bases were obtained for most of the potentially novel species. A total of 67 bacterial species or phylotypes were detected, 25 of which have not yet been grown in vitro. Nineteen of the species or phylotypes, including Propionibacterium acnes, Staphylococcus spp., and the opportunistic pathogens Stenotrophomonas maltophilia and Ochrobactrum anthropi were detected in more than one subject. Other known species that were detected included Achromobacter spp., Afipia spp., Brevundimonas diminuta, Capnocytophaga spp., Cardiobacterium sp., Eikenella corrodens, Fusobacterium spp., Gemella haemoylsans, and Neisseria spp. Phylotypes that were unique to noma infections included those in the genera Eubacterium, Flavobacterium, Kocuria, Microbacterium, and Porphyromonas and the related Streptococcus salivarius and genera Sphingomonas and TREPONEMA: Since advanced noma lesions are infections open to the environment, it was not surprising to detect species not commonly associated with the oral cavity, e.g., from soil. Several species previously implicated as putative pathogens of noma, such as spirochetes and Fusobacterium spp., were detected in at least one subject. However, due to the limited number of available noma subjects, it was not possible at this time to associate specific species with the disease.

Bionota: Bacterias promotoras de crecimiento de microalgas: una nueva aproximación en el tratamiento de aguas residuales Microalgae growth-promoting bacteria: A novel approach in wastewater treatment

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Bashan Yoav

2003-12-01

Full Text Available Las bacterias promotoras de crecimiento en plantas (PGPB del género Azospirillum son conocidas porque mejo­ran el crecimiento de numerosas cosechas agrícolas; sin embargo, el presente trabajo pretende extender el uso de estas bacterias a "bacterias promotoras de crecimiento de microalgas" (MPGB para aumentar la capacidad de las microalgas de eliminar nutrientes de aguas residuales. La inoculación deliberada de las microalgas Chlorella spp. con PGPB de origen terrestre no ha sido reportada con anterioridad, tal vez debido al origen diferente de estos dos microorganismos. Al inmovilizar de manera conjunta Chlorella vulgaris y Azospirillum brasilense Cd en esferas de alginato, se obtuvo como resultado un aumento significativo en varios parámetros de crecimiento de la microalga, como el peso fresco y seco, el número total de células, el tamaño de las colonias de microalgas dentro de la esfera, el número de organismos por colonia y la concentración de pigmentos. Además, aumenta­ron los lípidos y la variedad de ácidos grasos. La microalga combinada con la MGPB tiene una mayor capacidad de eliminar amonio y fósforo tanto en agua residual sintética como en agua residual doméstica. Actualmente se ha estado experimentando con otras PGPB (Flavobacterium sp. Azospirillum sp. y Azotobacter sp. para propósitos acuícolas; por ejemplo aumentar el crecimiento de fitoplancton utilizado en el cultivo de carpas y estabilizar cultivos masivos de microalgas marinas utilizadas como alimento para organismos marinos, todo esto con resul­tados promisorios. Si bien el efecto de las PGPB en microorganismos acuáticos aún no ha sido suficientemente explorado, proponemos que la co-inmovilización de microalgas y bacterias promotoras de crecimiento es un medio efectivo para aumentar la población microalgal y también su capacidad de limpiar aguas residuales. Palabras clave: PGPB; microalgas; biotratamiento de aguas residuales; co

Analysis of the microbiota of refrigerated chopped parsley after treatments with a coating containing enterocin AS-48 or by high-hydrostatic pressure.

Science.gov (United States)

Grande Burgos, María José; López Aguayo, María Del Carmen; Pérez Pulido, Rubén; Galvez, Antonio; Lucas, Rosario

2017-09-01

Parsley can be implicated in foodborne illness, yet chopped parsley is used as an ingredient or garnish for multiple dishes. The aim of the present study was to determine the effect of two different treatments on the bacterial diversity of parsley: (i) coating with a pectin-EDTA solution containing the circular bacteriocin enterocin AS-48, and (ii) treatment by high hydrostatic pressure (HHP) at 600MPa for 8min. Control and treated parsley were stored in trays at 5°C for 10days. Both treatments reduced viable counts by 3.7 log cycles and retarded growth of survivors during storage. The bacterial diversity of the chopped parsley was studied by high throughput sequencing (Illumina Miseq). Bacterial diversity of control samples mainly consists of Proteobacteria (96.87%) belonging to genera Pseudomonas (69.12%), Rheinheimera (8.56%) and Pantoea (6.91%) among others. During storage, the relative abundance of Bacteroidetes (mainly Flavobacterium and Sphingobacterium) increased to 26.66%. Application of the pectin-bacteriocin-EDTA coating reduced the relative abundance of Proteobacteria (63.75%) and increased that of Firmicutes (34.70%). However, the relative abundances of certain groups such as Salmonella, Shigella and Acinetobacter increased at early storage times. Late storage was characterized by an increase in the relative abundance of Proteobacteria, mainly Pseudomonas. Upon application of HHP treatment, the relative abundance of Proteobacteria was reduced (85.88%) while Actinobacteria increased (8.01%). During early storage of HHP-treated samples, the relative abundance of Firmicutes increased. Potentially-pathogenic bacteria (Shigella) only increased in relative abundance by the end of storage. Results of the present study indicate that the two treatments had different effects on the bacterial diversity of parsley. The HHP treatment provided a safer product, since no potentially-pathogenic bacteria were detected until the end of the storage period. Copyright �

Taxonomic profiles in metagenomic analyses of free-living microbial communities in the Ofunato Bay

KAUST Repository

Reza, Md. Shaheed; Kobiyama, Atsushi; Yamada, Yuichiro; Ikeda, Yuri; Ikeda, Daisuke; Mizusawa, Nanami; Ikeo, Kazuho; Sato, Shigeru; Ogata, Takehiko; Jimbo, Mitsuru; Kudo, Toshiaki; Kaga, Shinnosuke; Watanabe, Shiho; Naiki, Kimiaki; Kaga, Yoshimasa; Mineta, Katsuhiko; Bajic, Vladimir B.; Gojobori, Takashi; Watabe, Shugo

2018-01-01

The Ofunato Bay in Iwate Prefecture, Japan is a deep coastal bay located at the center of the Sanriku Rias coast and considered an economically and environmentally important asset. Here, we describe the first whole genome sequencing (WGS) study on the microbial community of the bay, where surface water samples were collected from three stations along its length to cover the entire bay; we preliminarily sequenced a 0.2 μm filter fraction among sequentially size-fractionated samples of 20.0, 5.0, 0.8 and 0.2 μm filters, targeting the free-living fraction only. From the 0.27–0.34 Gb WGS library, 0.9 × 106–1.2 × 106 reads from three sampling stations revealed 29 bacterial phyla (~80% of assigned reads), 3 archaeal phyla (~4%) and 59 eukaryotic phyla (~15%). Microbial diversity obtained from the WGS approach was compared with 16S rRNA gene results by mining WGS metagenomes, and we found similar estimates. The most frequently recovered bacterial sequences were Proteobacteria, predominantly comprised of 18.0–19.6% Planktomarina (Family Rhodobacteraceae) and 13.7–17.5% Candidatus Pelagibacter (Family Pelagibacterales). Other dominant bacterial genera, including Polaribacter (3.5–6.1%), Flavobacterium (1.8–2.6%), Sphingobacterium (1.4–1.6%) and Cellulophaga (1.4–2.0%), were members of Bacteroidetes and likely associated with the degradation and turnover of organic matter. The Marine Group I Archaea Nitrosopumilus was also detected. Remarkably, eukaryotic green alga Bathycoccus, Ostreococcus and Micromonas accounted for 8.8–15.2%, 3.6–4.9% and 2.1–3.1% of total read counts, respectively, highlighting their potential roles in the phytoplankton bloom after winter mixing.

Biodiversity of Rock Varnish at Yungay, Atacama Desert, Chile

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Kuhlman, K.; Venkat, P.; La Duc, M.; Kuhlman, G.; McKay, C.

2007-12-01

Rock varnish is a very slow-growing nanostratigraphic coating consisting of approximately 70% clay and 30% iron and manganese oxides of fine-grained clay minerals rich in manganese and iron oxides, which forms on the surfaces of rocks in most semi-arid to hyper-arid climates. Rock varnish has even been postulated to exist on Mars based on surface imagery from several landed missions, and is considered a potential biomarker. However, the mechanism of varnish nucleation and growth remains unknown. Whether or not microbes are involved in the nucleation and growth of rock varnish, the detection of microbes using cultivation or cultivation- independent techniques has demonstrated that varnish provides a microhabitat for microbes. We hypothesized that rock varnish in the Mars-like Yungay region of the Atacama Desert may provide such a microhabitat for microbial life where none has been found to date in the surface soil (< 1 cm). The presence of microbes was investigated using adenosine triphosphate (ATP) assay techniques and culture-independent biomolecular methods. High levels of both total and intracellular ATP were associated with the rock varnish while negligible ATP was found in the surrounding surface soil, suggesting that viable organisms were present. Total DNA was extracted from ground varnish and surrounding surface soil and subjected to trifurcate polymerase chain reactions (PCR). No DNA was recovered from the soil. Amplicons were used to generate ribosomal DNA (rDNA) clone libraries, which suggest the presence of numerous phylogenetically distinct microorganisms in eight Eubacterial clades, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Cytophaga-Flavobacterium- Bacteroides (CFB), Chloroflexi (green non-sulfur bacteria (GNS)), Gemmatimonadetes, Actinobacteria and Cyanobacteria. The diversity of bacteria found and presence of cyanobacteria suggests that rock varnish provides a niche environment for a cryptoendolithic microbial community where

Taxonomic profiles in metagenomic analyses of free-living microbial communities in the Ofunato Bay

KAUST Repository

Reza, Md. Shaheed

2018-04-27

The Ofunato Bay in Iwate Prefecture, Japan is a deep coastal bay located at the center of the Sanriku Rias coast and considered an economically and environmentally important asset. Here, we describe the first whole genome sequencing (WGS) study on the microbial community of the bay, where surface water samples were collected from three stations along its length to cover the entire bay; we preliminarily sequenced a 0.2 μm filter fraction among sequentially size-fractionated samples of 20.0, 5.0, 0.8 and 0.2 μm filters, targeting the free-living fraction only. From the 0.27–0.34 Gb WGS library, 0.9 × 106–1.2 × 106 reads from three sampling stations revealed 29 bacterial phyla (~80% of assigned reads), 3 archaeal phyla (~4%) and 59 eukaryotic phyla (~15%). Microbial diversity obtained from the WGS approach was compared with 16S rRNA gene results by mining WGS metagenomes, and we found similar estimates. The most frequently recovered bacterial sequences were Proteobacteria, predominantly comprised of 18.0–19.6% Planktomarina (Family Rhodobacteraceae) and 13.7–17.5% Candidatus Pelagibacter (Family Pelagibacterales). Other dominant bacterial genera, including Polaribacter (3.5–6.1%), Flavobacterium (1.8–2.6%), Sphingobacterium (1.4–1.6%) and Cellulophaga (1.4–2.0%), were members of Bacteroidetes and likely associated with the degradation and turnover of organic matter. The Marine Group I Archaea Nitrosopumilus was also detected. Remarkably, eukaryotic green alga Bathycoccus, Ostreococcus and Micromonas accounted for 8.8–15.2%, 3.6–4.9% and 2.1–3.1% of total read counts, respectively, highlighting their potential roles in the phytoplankton bloom after winter mixing.

Microalgae growth-promoting bacteria: A novel approach in wastewater treatment

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Luz E. de-Bashan

2003-07-01

Full Text Available Plant growth-promoting bacteria (PGPB from the genus Azospirillum are known to enhance the growth of numerous agricultural crops. The use of these bacteria is proposed as "micro-algae-growth promoting bacteria" (MGPB for enhancing freshwater micro-algae Chlorella vulgaris and C. sorokiniana capadty to clean polluted water. The deliberate inoculation of Chlorella sp. with a terrestrial PGPB has not been reported prior to these studies, perhaps because of the different origin of the two micro-organisms. Chlorella spp. is not known to harbour any plant growth-promoting bacteria and Azospirillum sp. is rarely used for inoculation in aquatic environments. Co-immobilisation of C. vulgaris and A. brasilense Cd in small alginate beads resulted in significant increases in numerous micro-algae growth parameters. Dry and fresh weight, total number of cells, micro-algal cluster (colonies size within the bead, number of micro-algal cells per cluster and micro-algal pigments levels significantly increased. Lipids and the variety of fatty adds also significantly increased, as did the combination of micro-algae. MGPB had superior capacity for removing ammonium and phosphorus from polluted synthetic and municipal wastewaters than the micro-algae by itself. Other PGPB (i.e. Flavobacterium sp. Azospirillum sp. and Azotobacter sp. are currently being tested in aquaculture; carp farming using enhanced phytoplankton growth and stabilising mass marine micro-algae culture for use as feed for marine organisms are both retuming promising results. This aspect of PGPB effect on water micro-organisms is currently in its infancy. We pro pose that co-immobilising micro-algae and plant growth-promoting bacteria represent an effective means of increasing micro-algal populations and also their capacity for cleaning polluted water. Key words: PGPB; micro-algae; wastewater treatment; co-immobilised

High-Level Expression of a Thermally Stable Alginate Lyase Using Pichia pastoris, Characterization and Application in Producing Brown Alginate Oligosaccharide

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Haifeng Li

2018-05-01

Full Text Available An alginate lyase encoding gene sagl from Flavobacterium sp. H63 was codon optimized and recombinantly expressed at high level in P.pastoris through high cell-density fermentation. The highest yield of recombinant enzyme of sagl (rSAGL in yeast culture supernatant reached 226.4 μg/mL (915.5 U/mL. This was the highest yield record of recombinant expression of alginate lyase so far. The rSAGL was confirmed as a partially glycosylated protein through EndoH digestion. The optimal reaction temperature and pH of this enzyme were 45 °C and 7.5; 80 mM K+ ions could improve the catalytic activity of the enzyme by 244% at most. rSAGL was a thermal stable enzyme with T5015 of 57–58 °C and T5030 of 53–54 °C. Its thermal stability was better than any known alginate lyase. In 100 mM phosphate buffer of pH 6.0, rSAGL could retain 98.8% of the initial activity after incubation at 50 °C for 2 h. Furthermore, it could retain 61.6% of the initial activity after 48 h. The specific activity of the purified rSAGL produced by P. pastoris attained 4044 U/mg protein, which was the second highest record of alginate lyase so far. When the crude enzyme of the rSAGL was directly used in transformation of sodium alginate with 40 g/L, 97.2% of the substrate was transformed to di, tri, tetra brown alginate oligosaccharide after 32 h of incubation at 50 °C, and the final concentration of reducing sugar in mixture reached 9.51 g/L. This is the first report of high-level expression of thermally stable alginate lyase using P. pastoris system.

Analysis of Bacterial Diversity in Different Heavy Oil Wells of a Reservoir in South Oman with Alkaline pH

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Biji Shibulal

2018-01-01

Full Text Available The identification of potential hydrocarbon utilizing bacteria is an essential requirement in microbial enhanced oil recovery (MEOR. Molecular approaches like proteomic and genomic characterization of the isolates are replacing the traditional method of identification with systemic classification. Genotypic profiling of the isolates includes fingerprint or pattern-based technique and sequence-based technique. Understanding community structure and dynamics is essential for studying diversity profiles and is challenging in the case of microbial analysis. The present study aims to understand the bacterial community composition from different heavy oil contaminated soil samples collected from geographically related oil well areas in Oman and to identify spore-forming hydrocarbon utilizing cultivable bacteria. V4 region of 16S rDNA gene was the target for Ion PGM™. A total of 825081 raw sequences were obtained from Ion torrent from all the 10 soil samples. The species richness and evenness were found to be moderate in all the samples with four main phyla, Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria, the most abundant being Firmicutes. Bacillus sp. ubiquitously dominated in all samples followed by Paenibacillus, which was followed by Brevibacillus, Planococcus, and Flavobacterium. Principal Coordinate Analysis (PCoA and UPGMA dendrogram clustered the 10 soil samples into four main groups. Weighted UniFrac significance test determined that there was significant difference in the communities present in soil samples examined. It can be concluded that the microbial community was different in all the 10 soil samples with Bacillus and Paenibacillus sp. as predominating genus. The 16S rDNA sequencing of cultivable spore-forming bacteria identified the hydrocarbon utilizing bacteria as Bacillus and Paenibacillus sp. and the nucleotide sequences were submitted to NCBI GenBank under accession numbers KP119097–KP119115. Bacillus and

Improved biostability assessment of drinking water with a suite of test methods at a water supply treating eutrophic lake water.

Science.gov (United States)

van der Kooij, Dick; Martijn, Bram; Schaap, Peter G; Hoogenboezem, Wim; Veenendaal, Harm R; van der Wielen, Paul W J J

2015-12-15

Assessment of drinking-water biostability is generally based on measuring bacterial growth in short-term batch tests. However, microbial growth in the distribution system is affected by multiple interactions between water, biofilms and sediments. Therefore a diversity of test methods was applied to characterize the biostability of drinking water distributed without disinfectant residual at a surface-water supply. This drinking water complied with the standards for the heterotrophic plate count and coliforms, but aeromonads periodically exceeded the regulatory limit (1000 CFU 100 mL(-1)). Compounds promoting growth of the biopolymer-utilizing Flavobacterium johnsoniae strain A3 accounted for c. 21% of the easily assimilable organic carbon (AOC) concentration (17 ± 2 μg C L(-1)) determined by growth of pure cultures in the water after granular activated-carbon filtration (GACF). Growth of the indigenous bacteria measured as adenosine tri-phosphate in water samples incubated at 25 °C confirmed the low AOC in the GACF but revealed the presence of compounds promoting growth after more than one week of incubation. Furthermore, the concentration of particulate organic carbon in the GACF (83 ± 42 μg C L(-1), including 65% carbohydrates) exceeded the AOC concentration. The increased biomass accumulation rate in the continuous biofouling monitor (CBM) at the distribution system reservoir demonstrated the presence of easily biodegradable by-products related to ClO2 dosage to the GACF and in the CBM at 42 km from the treatment plant an iron-associated biomass accumulation was observed. The various methods applied thus distinguished between easily assimilable compounds, biopolymers, slowly biodegradable compounds and biomass-accumulation potential, providing an improved assessment of the biostability of the water. Regrowth of aeromonads may be related to biomass-turnover processes in the distribution system, but establishment of quantitative relationships is needed for

Microbial Community Structure and Function Indicate the Severity of Chromium Contamination of the Yellow River

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Yaxin Pei

2018-01-01

Full Text Available The Yellow River is the most important water resource in northern China. In the recent past, heavy metal contamination has become severe due to industrial processes and other anthropogenic activities. In this study, riparian soil samples with varying levels of chromium (Cr pollution severity were collected along the Gansu industrial reach of the Yellow River, including samples from uncontaminated sites (XC, XGU, slightly contaminated sites (LJX, XGD, and heavily contaminated sites (CG, XG. The Cr concentrations of these samples varied from 83.83 mg⋅kg-1 (XGU to 506.58 mg⋅kg-1 (XG. The chromate [Cr (VI] reducing ability in the soils collected in this study followed the sequence of the heavily contaminated > slightly contaminated > the un-contaminated. Common Cr remediation genes chrA and yieF were detected in the XG and CG samples. qRT-PCR results showed that the expression of chrA was up-regulated four and threefold in XG and CG samples, respectively, whereas the expression of yieF was up-regulated 66- and 7-fold in the same samples after 30 min treatment with Cr (VI. The copy numbers of chrA and yieF didn’t change after 35 days incubation with Cr (VI. The microbial communities in the Cr contaminated sampling sites were different from those in the uncontaminated samples. Especially, the relative abundances of Firmicutes and Bacteroidetes were higher while Actinobacteria was lower in the contaminated group than uncontaminated group. Further, potential indicator species, related to Cr such as Cr-remediation genera (Geobacter, PSB-M-3, Flavobacterium, and Methanosarcina; the Cr-sensitive genera (Skermanella, Iamia, Arthrobacter, and Candidatus Nitrososphaera were also identified. These data revealed that Cr shifted microbial composition and function. Further, Cr (VI reducing ability could be related with the expression of Cr remediation genes.

Unique anthranilic acid chemistry facilitates profiling and characterization of Ser/Thr-linked sugar chains following hydrazinolysis.

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Anumula, Kalyan Rao

2008-02-01

A novel method for the analysis of Ser/Thr-linked sugar chains was made possible by the virtue of unique anthranilic acid (AA, 2-aminobenzoic acid [2AA]) chemistry for labeling carbohydrates in aqueous salt solutions (K. R. Anumula, Anal. Biochem. 350 (2006) 1-23). The protocol for profiling of Ser/Thr carbohydrates by hydrazinolysis was made simple by eliminating intermediary isolation steps involved in a sample preparation such as desalting and various chromatographic purification schemes. A 6-h hydrazinolysis was carried out at 60 degrees C for O-linked oligosaccharides and at 95 degrees C for total oligosaccharides (N-linked with some O-linked). Following evaporation of hydrazine (<10 min), the oligosaccharides were N-acetylated and derivatized with AA in the same reaction mixture containing salts. Presumably, the glycosyl-hydrazines/hydrazones present in the mixture did not interfere with AA labeling. Because AA is the most fluorescent and highly reactive tag for labeling carbohydrates, the procedures described are suitable for the analysis of a limited amount of samples ( approximately 5 microg) by the current high-resolution high-performance liquid chromatography (HPLC) methods. HPLC conditions developed for the separation of O-linked sugar chains based on size on an amide column were satisfactory for quantitative profiling and characterization. Common O-linked sugar chains found in fetuin, equine chorionic gonadotropin, and glycophorin can be analyzed in less than 50 min. In addition, these fast profiling methods were comparable to profiling by PNGase F (peptide N-glycosidase from Flavobacterium meningosepticum) digestion in terms of time, effort, and simplicity and also were highly reproducible for routine testing. The procedures for the release of sugar chains by hydrazinolysis at the microgram level, labeling with fluorescent tag AA, and profiling by HPLC should be useful in characterization of carbohydrates found in glycoproteins.

Biodegradation of international jet A-1 aviation fuel by microorganisms isolated from aircraft tank and joint hydrant storage systems.

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Itah, A Y; Brooks, A A; Ogar, B O; Okure, A B

2009-09-01

Microorganisms contaminating international Jet A-1 aircraft fuel and fuel preserved in Joint Hydrant Storage Tank (JHST) were isolated, characterized and identified. The isolates were Bacillus subtillis, Bacillus megaterium, Flavobacterium oderatum, Sarcina flava, Micrococcus varians, Pseudomonas aeruginosa, Bacillus licheniformis, Bacillus cereus and Bacillus brevis. Others included Candida tropicalis, Candida albicans, Saccharomyces estuari, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Aspergillus flavus, Aspergillus niger, Aspergillus fumigatus, Cladosporium resinae, Penicillium citrinum and Penicillium frequentans. The viable plate count of microorganisms in the Aircraft Tank ranged from 1.3 (+/-0.01) x 104 cfu/mL to 2.2 (+/-1.6) x 104 cfu/mL for bacteria and 102 cfu/mL to 1.68 (+/-0.32) x 103 cfu/mL for fungi. Total bacterial counts of 1.79 (+/-0.2) x 104 cfu/mL to 2.58 (+/-0.04) x 104 cfu/mL and total fungal count of 2.1 (+/-0.1) x 103 cfu/mL to 2.28 (+/-0.5) x 103 cfu/mL were obtained for JHST. Selected isolates were re-inoculated into filter sterilized aircraft fuels and biodegradation studies carried out. After 14 days incubation, Cladosporium resinae exhibited the highest degradation rate with a percentage weight loss of 66 followed by Candida albicans (60.6) while Penicillium citrinum was the least degrader with a weight loss of 41.6%. The ability of the isolates to utilize the fuel as their sole source of carbon and energy was examined and found to vary in growth profile between the isolates. The results imply that aviation fuel could be biodegraded by hydrocarbonoclastic microorganisms. To avert a possible deterioration of fuel quality during storage, fuel pipe clogging and failure, engine component damage, wing tank corrosion and aircraft disaster, efficient routine monitoring of aircraft fuel systems is advocated.

Increasing tetracycline concentrations on the performance and communities of mixed microalgae-bacteria photo-bioreactors

KAUST Repository

Xiong, Yanghui

2017-12-11

This study investigated the impact of varying concentrations of tetracycline on the performance of mixed microalgae-bacteria photo-bioreactors. Photo-bioreactors were assessed for their ability to remove carbon dioxide (CO2) from the biogas of anaerobic membrane bioreactor (anMBR), and nutrients from the anaerobic effluent. The varying concentrations of tetracycline had no impact on the removal of CO2 from biogas. 29% v/v of CO2 was completely removed to generate >20% v/v of oxygen (O2) in all reactors. Removal of nutrients and biomass was not affected at low concentrations of tetracycline (≤150μg/L), but 20mg/L of tetracycline lowered the biomass generation and removal efficiencies of phosphate. Conversely, high chlorophyll a and b content was observed at 20mg/L of tetracycline. High tetracycline level had no impact on the diversity of 18S rRNA gene-based microalgal communities but adversely affected the 16S rRNA gene-based microbial communities. Specifically, both Proteobacteria and Bacteroidetes phyla decreased in relative abundance but not phylum Chloroplast. Additionally, both nitrogen-fixing (e.g. Flavobacterium, unclassified Burkholderiales and unclassified Rhizobiaceae) and denitrifying groups (e.g. Hydrogenophaga spp.) were significantly reduced in relative abundance at high tetracycline concentration. Phosphate-accumulating microorganisms, Acinetobacter spp. and Pseudomonas spp. were similarly reduced upon exposure to high tetracycline concentration. Unclassified Comamonadaceae, however, increased in relative abundance, which correlated with an increase in the abundance of tetracycline resistance genes associated with efflux pump mechanism. Overall, the findings demonstrate that antibiotic concentrations in municipal wastewaters will not significantly affect the removal of nutrients by the mixed microalgae-bacteria photo-bioreactors. However, utilizing such photo-bioreactors as a polishing step for anMBRs that treat wastewaters with high tetracycline

Chestnut green waste composting for sustainable forest management: Microbiota dynamics and impact on plant disease control.

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Ventorino, Valeria; Parillo, Rita; Testa, Antonino; Viscardi, Sharon; Espresso, Francesco; Pepe, Olimpia

2016-01-15

Making compost from chestnut lignocellulosic waste is a possible sustainable management strategy for forests that employs a high-quality renewable organic resource. Characterization of the microbiota involved in composting is essential to better understand the entire process as well as the properties of the final product. Therefore, this study investigated the microbial communities involved in the composting of chestnut residues obtained from tree cleaning and pruning. The culture-independent approach taken highlighted the fact that the microbiota varied only slightly during the process, with the exception of those of the starting substrate and mature compost. The statistical analysis indicated that most of the bacterial and fungal species in the chestnut compost persisted during composting. The dominant microbial population detected during the process belonged to genera known to degrade recalcitrant lignocellulosic materials. Specifically, we identified fungal genera, such as Penicillium, Fusarium, Cladosporium, Aspergillus and Mucor, and prokaryotic species affiliated with Bacilli, Actinobacteria, Flavobacteria and γ-Proteobacteria. The suppressive properties of compost supplements for the biocontrol of Sclerotinia minor and Rhizoctonia solani were also investigated. Compared to pure substrate, the addition of compost to the peat-based growth substrates resulted in a significant reduction of disease in tomato plants of up to 70 % or 51 % in the presence of Sclerotinia minor or Rhizoctonia solani, respectively. The obtained results were related to the presence of putative bio-control agents and plant growth-promoting rhizobacteria belonging to the genera Azotobacter, Pseudomonas, Stenotrophomonas, Bacillus, Flavobacterium, Streptomyces and Actinomyces in the chestnut compost. The composting of chestnut waste may represent a sustainable agricultural practice for disposing of lignocellulosic waste by transforming it into green waste compost that can be used to

Prebiotic Supplementation Has Only Minimal Effects on Growth Efficiency, Intestinal Health and Disease Resistance of Westslope Cutthroat Trout Oncorhynchus clarkii lewisi Fed 30% Soybean Meal

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Wendy M. Sealey

2015-08-01

Full Text Available Prebiotics have successfully been used to prevent infectious diseases in aquaculture and there is an increasing amount of literature that suggests that these products can also improve alternative protein utilization and digestion. Therefore, the objective of this study was to examine whether prebiotic supplementation increased the growth efficiency, intestinal health and disease resistance of cutthroat trout fed a high level of dietary soybean meal. To achieve this objective, juvenile Westslope cutthroat trout (Oncorhynchus clarkii lewisi were fed a practical type formulation with 0 or 30% dietary soybean meal with or without the commercial prebiotic (Grobiotic-A prior to experimental exposure to Flavobacterium psychrophilum. Juvenile Westslope cutthroat trout (initial weight 7.8 g/fish ± standard deviation of 0.5 g were stocked at 30 fish/tank in 75 L tanks with six replicate tanks per diet and fed their respective diets for 20 weeks. Final weights of Westslope cutthroat trout were affected by neither dietary soybean meal inclusion level (P=0.9582 nor prebiotic inclusion (P=0.9348 and no interaction was observed (P=0.1242. Feed conversion ratios were similarly not affected by soybean meal level (P=0.4895, prebiotic inclusion (P=0.3258 or their interaction (P=0.1478. Histological examination of the distal intestine of Westslope cutthroat trout demonstrated increases in inflammation due to both increased soybean meal inclusion level (P=0.0038 and prebiotic inclusion (P=0.0327 without significant interaction (P=0.3370. Feeding dietary soybean meal level at 30% increased mortality of F.psychrophilum cohabitation challenged Westslope cutthroat trout (P=0.0345 while prebiotic inclusion tended to decrease mortality (P=0.0671. These results indicate that subclinical alterations in intestinal inflammation levels due to high dietary inclusion levels of soybean meal could predispose Westslope cutthroat trout to F.psychrophilum infection.

Prebiotic Supplementation has Only Minimal Effects on Growth Efficiency, Intestinal Health and Disease Resistance of Westslope Cutthroat Trout Oncorhynchus clarkii lewisi Fed 30% Soybean Meal.

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Sealey, Wendy M; Conley, Zachariah B; Bensley, Molly

2015-01-01

Prebiotics have successfully been used to prevent infectious diseases in aquaculture and there is an increasing amount of literature that suggests that these products can also improve alternative protein utilization and digestion. Therefore, the objective of this study was to examine whether prebiotic supplementation increased the growth efficiency, intestinal health, and disease resistance of cutthroat trout fed a high level of dietary soybean meal. To achieve this objective, juvenile Westslope cutthroat trout (Oncorhynchus clarkii lewisi) were fed a practical type formulation with 0 or 30% dietary soybean meal with or without the commercial prebiotic (Grobiotic-A) prior to experimental exposure to Flavobacterium psychrophilum. Juvenile Westslope cutthroat trout (initial weight 7.8 g/fish ±SD of 0.5 g) were stocked at 30 fish/tank in 75 L tanks with six replicate tanks per diet and fed their respective diets for 20 weeks. Final weights of Westslope cutthroat trout were affected by neither dietary soybean meal inclusion level (P = 0.9582) nor prebiotic inclusion (P = 0.9348) and no interaction was observed (P = 0.1242). Feed conversion ratios were similarly not affected by soybean meal level (P = 0.4895), prebiotic inclusion (P = 0.3258) or their interaction (P = 0.1478). Histological examination of the distal intestine of Westslope cutthroat trout demonstrated increases in inflammation due to both increased soybean meal inclusion level (P = 0.0038) and prebiotic inclusion (P = 0.0327) without significant interaction (P = 0.3370). Feeding dietary soybean meal level at 30% increased mortality of F. psychrophilum cohabitation challenged Westslope cutthroat trout (P = 0.0345) while prebiotic inclusion tended to decrease mortality (P = 0.0671). These results indicate that subclinical alterations in intestinal inflammation levels due to high dietary inclusion levels of soybean meal could predispose Westslope cutthroat

Degradation of organic pollutants by methane grown microbial consortia.

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Hesselsoe, Martin; Boysen, Susanne; Iversen, Niels; Jørgensen, Lars; Murrell, J Colin; McDonald, Ian; Radajewski, Stefan; Thestrup, Helle; Roslev, Peter

2005-10-01

Microbial consortia were enriched from various environmental samples with methane as the sole carbon and energy source. Selected consortia that showed a capacity for co-oxidation of naphthalene were screened for their ability to degrade methyl-tert-butyl-ether (MTBE), phthalic acid esters (PAE), benzene, xylene and toluene (BTX). MTBE was not removed within 24 h by any of the consortia examined. One consortium enriched from activated sludge ("AAE-A2"), degraded PAE, including (butyl-benzyl)phthalate (BBP), and di-(butyl)phthalate (DBP). PAE have not previously been described as substrates for methanotrophic consortia. The apparent Km and Vmax for DBP degradation by AAE-A2 at 20 degrees C was 3.1 +/- 1.2 mg l(-1) and 8.7 +/- 1.1 mg DBP (g protein x h)(-1), respectively. AAE-A2 also showed fast degradation of BTX (230 +/- 30 nmol benzene (mg protein x h)(-1) at 20 degrees C). Additionally, AAE-A2 degraded benzene continuously for 2 weeks. In contrast, a pure culture of the methanotroph Methylosinus trichosporium OB3b ceased benzene degradation after only 2 days. Experiments with methane mono-oxygenase inhibitors or competitive substrates suggested that BTX degradation was carried out by methane-oxidizing bacteria in the consortium, whereas the degradation of PAE was carried out by non-methanotrophic bacteria co-existing with methanotrophs. The composition of the consortium (AAE-A2) based on polar lipid fatty acid (PLFA) profiles showed dominance of type II methanotrophs (83-92% of biomass). Phylogeny based on a 16S-rRNA gene clone library revealed that the dominating methanotrophs belonged to Methylosinus/Methylocystis spp. and that members of at least 4 different non-methanotrophic genera were present (Pseudomonas, Flavobacterium, Janthinobacterium and Rubivivax).

The link of feast-phase dissolved oxygen (DO) with substrate competition and microbial selection in PHA production.

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Wang, Xiaofei; Oehmen, Adrian; Freitas, Elisabete B; Carvalho, Gilda; Reis, Maria A M

2017-04-01

Polyhydroxyalkanoates (PHAs) are biobased and biodegradable polyesters with the potential to replace conventional plastics. Aeration requires large amounts of energy in PHA production by mixed microbial cultures (MMCs), particularly during the feast phase due to substrate uptake. The objective of this study was to investigate the impact of DO concentrations on microbial selection, substrate competition and PHA production performance by MMCs. This represents the first study investigating DO impact on PHA production while feeding the multiple volatile fatty acids (VFAs) typically encountered in real fermented feedstocks, as well as the substrate preferences at different DO levels. Efficient microbial cultures were enriched under both high (3.47 ± 1.12 mg/L) and low (0.86 ± 0.50 mg/L) DO conditions in the feast phase containing mostly the same populations but with different relative abundance. The most abundant microorganisms in the two MMCs were Plasticicumulans, Zoogloea, Paracoccus, and Flavobacterium. Butyrate and valerate were found to be the preferred substrates as compared to acetate and propionate regardless of DO concentrations. In the accumulation step, the PHA storage capacity and yield were less affected by the change of DO levels when applying the culture selected under low DO in the feast phase (PHA storage capacity >60% and yield > 0.9 Cmol PHA/Cmol VFA). A high DO level is required for maximal PHA accumulation rates with the four VFAs (acetate, propionate, butyrate and valerate) present, due to the lower specific uptake rates of acetate and propionate under low DO conditions. However, butyrate and valerate specific uptake rates were less impacted by DO levels and hence low DO for PHA accumulation may be effective when feed is composed of these substrates only. Copyright © 2017 Elsevier Ltd. All rights reserved.

Covariance of bacterioplankton composition and environmental variables in a temperate delta system

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Stepanauskas, R.; Moran, M.A.; Bergamaschi, B.A.; Hollibaugh, J.T.

2003-01-01

We examined seasonal and spatial variation in bacterioplankton composition in the Sacramento-San Joaquin River Delta (CA) using terminal restriction fragment length polymorphism (T-RFLP) analysis. Cloned 16S rRNA genes from this system were used for putative identification of taxa dominating the T-RFLP profiles. Both cloning and T-RFLP analysis indicated that Actinobacteria, Verrucomicrobia, Cytophaga-Flavobacterium and Proteobacteria were the most abundant bacterioplankton groups in the Delta. Despite the broad variety of sampled habitats (deep water channels, lakes, marshes, agricultural drains, freshwater and brackish areas), and the spatial and temporal differences in hydrology, temperature and water chemistry among the sampling campaigns, T-RFLP electropherograms from all samples were similar, indicating that the same bacterioplankton phylotypes dominated in the various habitats of the Delta throughout the year. However, principal component analysis (PCA) and partial least-squares regression (PLS) of T-RFLP profiles revealed consistent grouping of samples on a seasonal, but not a spatial, basis. ??-Proteobacteria related to Ralstonia, Actinobacteria related to Microthrix, and ??-Proteobacteria identical to the environmental Clone LD12 had the highest relative abundance in summer/fall T-RFLP profiles and were associated with low river flow, high pH, and a number of optical and chemical characteristics of dissolved organic carbon (DOC) indicative of an increased proportion of phytoplankton-produced organic material as opposed to allochthonous, terrestrially derived organic material. On the other hand, Geobacter-related ??-Proteobacteria showed a relative increase in abundance in T-RFLP analysis during winter/spring, and probably were washed out from watershed soils or sediment. Various phylotypes associated with the same phylogenetic division, based on tentative identification of T-RFLP fragments, exhibited diverse seasonal patterns, suggesting that ecological

Samanlarda Biyolojik Muamelelerle Lignoselüloz Kompleksin Sindirilebilirliğinin Artırılması

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Aydan Atalar

2017-12-01

Full Text Available Samanların hücre duvarında bulunan lignoselüloz kompleksin parçalanması ile selüloz ve hemiselüloz gibi rumen fermantasyonuyla sindirilebilir fraksiyonlar elde edilir. Rumende sindirilmeyen ligninin biyoteknolojik metotlarla parçalanarak samandan yararlanılabilirliğin artırılması son yıllarda hayvan beslemecilerin odak noktası olan alanlarından birisi olmuştur. Lignoselüloz kompleksin biyolojik metotlarla muamelesinde bakteriler, mantarlar ve bu mikroorganizmalardan elde edilen enzimler kullanılmaktadır. Bakteri muamelesinde Mycobacterium, Arthrobacter ve Flavobacterium türü bakterilerin lignini parçalayabilme özelliğinden yararlanılmaktadır. Enzim muamelesi etkili olmasına rağmen yüksek maliyet nedeniyle uygulamada yer bulamamıştır. Mantar muamelesinde beyaz, kahverengi ve yumuşak çürüme yapan 3 tür mantar kullanılmaktadır. Kahverengi çürükçül mantarlar tercihen selüloz ve hemiselüloza saldırır, ancak lignini parçalayamaz. Beyaz çürükçül mantarlar lignine saldırarak lignol bağları ve aromatik halkayı parçalarlar. Beyaz çürükçül mantarlar selülaz, ksilanaz gibi hidrolitik enzimlerle polisakkaritleri ve lignin peroksidaz ve lakkaz gibi oksidatif ligninolitik enzimlerle lignini parçalarlar. Lignoselülozik materyalleri en iyi parçalayabilen mikroorganizmaların mantarlar olması ve maliyetin düşük olması nedeniyle özellikle beyaz çürükçül mantarların uygulama potansiyeli bulunmaktadır. Bu bildiride biyoçeşitliliğin sağladığı avantajla biyolojik metotlarla samanların sindirilebilirliğinin artırılması tartışılacaktır.

Diversity, metabolic properties and arsenic mobilization potential of indigenous bacteria in arsenic contaminated groundwater of West Bengal, India.

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Paul, Dhiraj; Kazy, Sufia K; Gupta, Ashok K; Pal, Taraknath; Sar, Pinaki

2015-01-01

Arsenic (As) mobilization in alluvial aquifers is caused by a complex interplay of hydro-geo-microbiological activities. Nevertheless, diversity and biogeochemical significance of indigenous bacteria in Bengal Delta Plain are not well documented. We have deciphered bacterial community compositions and metabolic properties in As contaminated groundwater of West Bengal to define their role in As mobilization. Groundwater samples showed characteristic high As, low organic carbon and reducing property. Culture-independent and -dependent analyses revealed presence of diverse, yet near consistent community composition mostly represented by genera Pseudomonas, Flavobacterium, Brevundimonas, Polaromonas, Rhodococcus, Methyloversatilis and Methylotenera. Along with As-resistance and -reductase activities, abilities to metabolize a wide range carbon substrates including long chain and polyaromatic hydrocarbons and HCO3, As3+ as electron donor and As5+/Fe3+ as terminal electron acceptor during anaerobic growth were frequently observed within the cultivable bacteria. Genes encoding cytosolic As5+ reductase (arsC) and As3+ efflux/transporter [arsB and acr3(2)] were found to be more abundant than the dissimilatory As5+ reductase gene arrA. The observed metabolic characteristics showed a good agreement with the same derived from phylogenetic lineages of constituent populations. Selected bacterial strains incubated anaerobically over 300 days using natural orange sand of Pleistocene aquifer showed release of soluble As mostly as As3+ along with several other elements (Al, Fe, Mn, K, etc.). Together with the production of oxalic acid within the biotic microcosms, change in sediment composition and mineralogy indicated dissolution of orange sand coupled with As/Fe reduction. Presence of arsC gene, As5+ reductase activity and oxalic acid production by the bacteria were found to be closely related to their ability to mobilize sediment bound As. Overall observations suggest that

Influence of an oyster reef on development of the microbial heterotrophic community of an estuarine biofilm.

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Nocker, Andreas; Lepo, Joe E; Snyder, Richard A

2004-11-01

We characterized microbial biofilm communities developed over two very closely located but distinct benthic habitats in the Pensacola Bay estuary using two complementary cultivation-independent molecular techniques. Biofilms were grown for 7 days on glass slides held in racks 10 to 15 cm over an oyster reef and an adjacent muddy sand bottom. Total biomass and optical densities of dried biofilms showed dramatic differences for oyster reef versus non-oyster reef biofilms. This study assessed whether the observed spatial variation was reflected in the heterotrophic prokaryotic species composition. Genomic biofilm DNA from both locations was isolated and served as a template to amplify 16S rRNA genes with universal eubacterial primers. Fluorescently labeled PCR products were analyzed by terminal restriction fragment length polymorphism, creating a genetic fingerprint of the composition of the microbial communities. Unlabeled PCR products were cloned in order to construct a clone library of 16S rRNA genes. Amplified ribosomal DNA restriction analysis was used to screen and define ribotypes. Partial sequences from unique ribotypes were compared with existing database entries to identify species and to construct phylogenetic trees representative of community structures. A pronounced difference in species richness and evenness was observed at the two sites. The biofilm community structure from the oyster reef setting had greater evenness and species richness than the one from the muddy sand bottom. The vast majority of the bacteria in the oyster reef biofilm were related to members of the gamma- and delta-subdivisions of Proteobacteria, the Cytophaga-Flavobacterium -Bacteroides cluster, and the phyla Planctomyces and Holophaga-Acidobacterium. The same groups were also present in the biofilm harvested at the muddy sand bottom, with the difference that nearly half of the community consisted of representatives of the Planctomyces phylum. Total species richness was estimated

Biological properties of soils of former forest fires in Samosir Regency of North Sumatera

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D. Elfiati

2016-04-01

Full Text Available A study that was aimed to identify the impact of forest fires on the biological properties of soils was carried out at former forest fire areas in Samosir Regency of North Sumatera. Soil samples were collected from former forest fire areas of 2014, 2013, 2012, 2011, 2010. The composite soil samples were collected systematically using diagonal method as much as 5 points in each period of fire. The soil samples were taken at three plots measuring 20 x 20 m 0-20 cm depth. Soil biological properties observed were soil organic C content, total number of microbes, abundance of arbuscular mycorrhizal fungi, phosphate solubilizing microbes, and soil microbial activity. The results showed that organic C content ranged from 0.75 to 2.47% which included criteria for very low to moderate. Arbuscular mycorrhizal fungi spores were found belonging to the genus of Glomus and Acaulospora. Spore number increased with the fire period ranging from 45 spores (forest fire in 2014 to 152 spores (forest fire in 2010. The total number of microbes obtained ranged from 53.78 x 107 cfu/mL (forest fire in 2010 to 89.70 x107 cfu/mL (forest fire in 2013. It was found 29 isolates of phosphate solubilizing microbes that consisted of 14 bacterial isolates and 15 fungi isolates with densities ranging from 27.642 x105 cfu/mL (forest fires in 2014 to 97.776 x 105 cfu/ mL (forest fires in 2011. The isolates of phosphate solubilizing bacteria identified consisted of Pseudomonas, Flavobacterium, Staphylococcus, and Mycobacterium genus, whereas the isolates of phosphate solubilizing fungi obtained consisted of Aspergillus and Penicillium genus. Soil respiration ranged from 2.14 kg / day (forest fire in 2010 up to 3.71 kg / day (forest fire in 2013. The varied results were greatly influenced by the type or form of the fires and intensity of fires. In the study area the type or form of the fires were canopy fires with low intensity.

Occurrence and antibiotic susceptibility of fish bacteria isolated from Oreochromis niloticus (Nile tilapia and Clarias gariepinus (African catfish in Uganda

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S. P. Wamala

2018-02-01

Full Text Available Abstract The intention of this study was to identify the bacterial pathogens infecting Oreochromis niloticus (Nile tilapia and Clarias gariepinus (African catfish, and to establish the antibiotic susceptibility of fish bacteria in Uganda. A total of 288 fish samples from 40 fish farms (ponds, cages, and tanks and 8 wild water sites were aseptically collected and bacteria isolated from the head kidney, liver, brain and spleen. The isolates were identified by their morphological characteristics, conventional biochemical tests and Analytical Profile Index test kits. Antibiotic susceptibility of selected bacteria was determined by the Kirby-Bauer disc diffusion method. The following well-known fish pathogens were identified at a farm prevalence of; Aeromonas hydrophila (43.8%, Aeromonas sobria (20.8%, Edwardsiella tarda (8.3%, Flavobacterium spp. (4.2% and Streptococcus spp. (6.3%. Other bacteria with varying significance as fish pathogens were also identified including Plesiomonas shigelloides (25.0%, Chryseobacterium indoligenes (12.5%, Pseudomonas fluorescens (10.4%, Pseudomonas aeruginosa (4.2%, Pseudomonas stutzeri (2.1%, Vibrio cholerae (10.4%, Proteus spp. (6.3%, Citrobacter spp. (4.2%, Klebsiella spp. (4.2% Serratia marcescens (4.2%, Burkholderia cepacia (2.1%, Comamonas testosteroni (8.3% and Ralstonia picketti (2.1%. Aeromonas spp., Edwardsiella tarda and Streptococcus spp. were commonly isolated from diseased fish. Aeromonas spp. (n = 82 and Plesiomonas shigelloides (n = 73 were evaluated for antibiotic susceptibility. All isolates tested were susceptible to at-least ten (10 of the fourteen antibiotics evaluated. High levels of resistance were however expressed by all isolates to penicillin, oxacillin and ampicillin. This observed resistance is most probably intrinsic to those bacteria, suggesting minimal levels of acquired antibiotic resistance in fish bacteria from the study area. To our knowledge, this is the first study to

B cell signatures of BCWD-resistant and susceptible lines of rainbow trout: a shift towards more EBF-expressing progenitors and fewer mature B cells in resistant animals.

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Zwollo, Patty; Ray, Jocelyn C; Sestito, Michael; Kiernan, Elizabeth; Wiens, Gregory D; Kaattari, Steve; StJacques, Brittany; Epp, Lidia

2015-01-01

Bacterial cold water disease (BCWD) is a chronic disease of rainbow trout, and is caused by the Gram-negative bacterium Flavobacterium psychrophilum (Fp), a common aquaculture pathogen. The National Center for Cool and Cold Water Aquaculture has bred two genetic lines of rainbow trout: a line of Fp-resistant trout (ARS-Fp-R or R-line trout) and a line of susceptible trout (ARS-Fp-S, or S-line). Little is known about how phenotypic selection alters immune response parameters or how such changes relate to genetic disease resistance. Herein, we quantify interindividual variation in the distribution and abundance of B cell populations (B cell signatures) and examine differences between genetic lines of naive animals. There are limited trout-specific cell surface markers currently available to resolve B cell subpopulations and thus we developed an alternative approach based on detection of differentially expressed transcription factors and intracellular cytokines. B cell signatures were compared between R-line and S-line trout by flow cytometry using antibodies against transcription factors early B cell factor-1 (EBF1) and paired domain box protein Pax5, the pro-inflammatory cytokine IL-1β, and the immunoglobulin heavy chain mu. R-line trout had higher percentages of EBF(+) B myeloid/ progenitor and pre-B cells in PBL, anterior and posterior kidney tissues compared to S-line trout. The opposite pattern was detected in more mature B cell populations: R-line trout had lower percentages of both IgM(+) mature B cells and IgM-secreting cells in anterior kidney and PBL compared to S-line trout. In vitro LPS-activation studies of PBL and spleen cell cultures revealed no significant induction differences between R-line and S-line trout. Together, our findings suggest that selective resistance to BCWD may be associated with shifts in naive animal developmental lineage commitment that result in decreased B lymphopoiesis and increased myelopoiesis in BCWD resistant trout relative

OPTIMASI KONSENTRASI INOKULUM, RASIO C:N:P DAN pH PADA PROSES BIOREMEDIASI LIMBAH PENGILANGAN MINYAK BUMI MENGGUNAKAN KULTUR CAMPURAN

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Syukria Ikhsan Zam

2010-03-01

Full Text Available The purposes of this research were to obtain the best inoculum concentration, C:N:P ratio, and pH, and also to identify the ability of mixed culture of hydrocarbonoclastic bacteria in oil waste degradation. The isolats were used are Acinetobacter baumanni, Alcaligenes eutrophus, Bacillus sp1., Methylococcus capsulatus, Bacillus sp2., Morococcus sp., Pseudomonas diminuta, Xanthomonas albilineans, Bacillus cereus and Flavobacterium branchiophiia. Variation of inoculum concentrations were 10%, 15%, and 20% (v/v, C:N:P ratios were 100:10:1, 100:10:0,5, 100:5:1, and 100:5:0,5, and pH were 6,5, 7,0, 7,5. Observed parameters in optimization were Total Plate Count (TPC the culture every 24 hours, Total Petroleum Hydrocarbon (TPH and Chemical Oxygen Demand (COD examined at the end of the bioremediation period. Best optimization result then analyzed with GC/MS. Optimization result indicated the best inoculum concentration was 10% with TPH degradation 61,79% and COD slope 61,75%. It is assumed that the low value of TPH degradation and COD slope at 15% and 20% inoculum concentration were caused by competition inside the bacterial population at that high inoculum concentration. The competition result in low growth and degradation. C:N:P ratio was 100:5:1 with TPH degradation 66,55% and COD slope 85,18%. It is assumed that the C:N:P ratio is equal, so it can enhance the bioremediation procces. The best pH was 7,5 with TPH degradation 73,24% and COD slope 86,28%. The process at the optimum conditions using inoculum as a mixed culture enhanced the bioremediation process with the result as follows, TPH degradation 93,06%, COD 90,73% for treatment. The chromatogram indicated that total hydrocarbon compound from nC9 – nC32 have been degraded by 43,413% – 63,117%. A good result of bioremediation was obtained from mixed culture inoculum at 10% concentration, C:N:P ratio of 100:5:1, and pH 7,5.

Culture dependent and independent analysis of bacterial communities associated with commercial salad leaf vegetables.

Science.gov (United States)

Jackson, Colin R; Randolph, Kevin C; Osborn, Shelly L; Tyler, Heather L

2013-12-01

Plants harbor a diverse bacterial community, both as epiphytes on the plant surface and as endophytes within plant tissue. While some plant-associated bacteria act as plant pathogens or promote plant growth, others may be human pathogens. The aim of the current study was to determine the bacterial community composition of organic and conventionally grown leafy salad vegetables at the point of consumption using both culture-dependent and culture-independent methods. Total culturable bacteria on salad vegetables ranged from 8.0 × 10(3) to 5.5 × 10(8) CFU g(-1). The number of culturable endophytic bacteria from surface sterilized plants was significantly lower, ranging from 2.2 × 10(3) to 5.8 × 10(5) CFU g(-1). Cultured isolates belonged to six major bacterial phyla, and included representatives of Pseudomonas, Pantoea, Chryseobacterium, and Flavobacterium. Eleven different phyla and subphyla were identified by culture-independent pyrosequencing, with Gammaproteobacteria, Betaproteobacteria, and Bacteroidetes being the most dominant lineages. Other bacterial lineages identified (e.g. Firmicutes, Alphaproteobacteria, Acidobacteria, and Actinobacteria) typically represented less than 1% of sequences obtained. At the genus level, sequences classified as Pseudomonas were identified in all samples and this was often the most prevalent genus. Ralstonia sequences made up a greater portion of the community in surface sterilized than non-surface sterilized samples, indicating that it was largely endophytic, while Acinetobacter sequences appeared to be primarily associated with the leaf surface. Analysis of molecular variance indicated there were no significant differences in bacterial community composition between organic versus conventionally grown, or surface-sterilized versus non-sterilized leaf vegetables. While culture-independent pyrosequencing identified significantly more bacterial taxa, the dominant taxa from pyrosequence data were also detected by traditional

Biological phosphorus removal during high-rate, low-temperature, anaerobic digestion of wastewater

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Ciara eKeating

2016-03-01

Full Text Available We report, for the first time, extensive biologically-mediated phosphate removal from wastewater during high-rate anaerobic digestion (AD. A hybrid sludge bed/fixed-film (packed pumice stone reactor was employed for low-temperature (12°C anaerobic treatment of synthetic sewage wastewater. Successful phosphate removal from the wastewater (up to 78% of influent phosphate was observed, mediated by biofilms in the reactor. Scanning electron microscopy and energy dispersive X-ray analysis revealed the accumulation of elemental phosphorus (~2% within the sludge bed and fixed-film biofilms. 4’, 6-diamidino-2-phenylindole (DAPI staining indicated phosphorus accumulation was biological in nature and mediated through the formation of intracellular inorganic polyphosphate (polyP granules within these biofilms. DAPI staining further indicated that polyP accumulation was rarely associated with free cells. Efficient and consistent chemical oxygen demand (COD removal was recorded, throughout the 732-day trial, at applied organic loading rates between 0.4-1.5 kg COD m-3 d-1 and hydraulic retention times of 8-24 hours, while phosphate removal efficiency ranged from 28-78% on average per phase. Analysis of protein hydrolysis kinetics and the methanogenic activity profiles of the biomass revealed the development, at 12˚C, of active hydrolytic and methanogenic populations. Temporal microbial changes were monitored using Illumina Miseq analysis of bacterial and archaeal 16S rRNA gene sequences. The dominant bacterial phyla present in the biomass at the conclusion of the trial were the Proteobacteria and Firmicutes and the dominant archaeal genus was Methanosaeta. Trichococcus and Flavobacterium populations, previously associated with low temperature protein degradation, developed in the reactor biomass. The presence of previously characterised polyphosphate accumulating organisms (PAOs such as Rhodocyclus, Chromatiales, Actinobacter and Acinetobacter was

Microbial communities and fecal indicator bacteria associated with Cladophora mats on beach sites along Lake Michigan shores.

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Olapade, Ola A; Depas, Morgan M; Jensen, Erika T; McLellan, Sandra L

2006-03-01

A high biomasses of Cladophora, a filamentous green alga, is found mainly during the summer along the shores of Lake Michigan. In this study, the abundance and persistence of the fecal indicator bacterium Escherichia coli and sulfate-reducing bacteria (SRB) on Cladophora mats collected at Lake Michigan beaches were evaluated using both culture-based and molecular analyses. Additionally, 16S rRNA gene cloning and sequencing were used to examine the bacterial community composition. Overall, E. coli was detected in all 63 samples obtained from 11 sites, and the average levels at most beaches ranged from 2,700 CFU/100 g (wet weight) of Cladophora to 7,500 CFU/100 g of Cladophora. However, three beaches were found to have site average E. coli densities of 12,800, 21,130, and 27,950 CFU/100 g of Cladophora. The E. coli levels in the lake water collected at the same time from these three sites were less than the recommended U.S. Environmental Protection Agency limit, 235 CFU/100 ml. E. coli also persisted on Cladophora mats in microcosms at room temperature for more than 7 days, and in some experiments it persisted for as long as 28 days. The SRB densities on Cladophora mats were relatively high, ranging from 4.4x10(6) cells/g (6.64 log CFU/g) to 5.73x10(6) cells/g (6.76 log CFU/g) and accounting for between 20% and 27% of the total bacterial counts. Partial sequences of the 16S rRNA gene clones revealed a phylogenetically diverse community, in which the Cytophaga-Flavobacterium-Bacteroides cluster and the low-G+C-content gram-positive bacteria were the dominant organisms, accounting for 40% and 12.8%, respectively, of the total clone library. These results further reveal the potential public health and ecological significance of Cladophora mats that are commonly found along the shoreline of Lake Michigan, especially with regard to the potential to harbor microorganisms associated with fecal pollution and odor-causing bacteria.

Acute effects of TiO2 nanomaterials on the viability and taxonomic composition of aquatic bacterial communities assessed via high-throughput screening and next generation sequencing.

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Chu Thi Thanh Binh

Full Text Available The nanotechnology industry is growing rapidly, leading to concerns about the potential ecological consequences of the release of engineered nanomaterials (ENMs to the environment. One challenge of assessing the ecological risks of ENMs is the incredible diversity of ENMs currently available and the rapid pace at which new ENMs are being developed. High-throughput screening (HTS is a popular approach to assessing ENM cytotoxicity that offers the opportunity to rapidly test in parallel a wide range of ENMs at multiple concentrations. However, current HTS approaches generally test one cell type at a time, which limits their ability to predict responses of complex microbial communities. In this study toxicity screening via a HTS platform was used in combination with next generation sequencing (NGS to assess responses of bacterial communities from two aquatic habitats, Lake Michigan (LM and the Chicago River (CR, to short-term exposure in their native waters to several commercial TiO2 nanomaterials under simulated solar irradiation. Results demonstrate that bacterial communities from LM and CR differed in their sensitivity to nano-TiO2, with the community from CR being more resistant. NGS analysis revealed that the composition of the bacterial communities from LM and CR were significantly altered by exposure to nano-TiO2, including decreases in overall bacterial diversity, decreases in the relative abundance of Actinomycetales, Sphingobacteriales, Limnohabitans, and Flavobacterium, and a significant increase in Limnobacter. These results suggest that the release of nano-TiO2 to the environment has the potential to alter the composition of aquatic bacterial communities, which could have implications for the stability and function of aquatic ecosystems. The novel combination of HTS and NGS described in this study represents a major advance over current methods for assessing ENM ecotoxicity because the relative toxicities of multiple ENMs to thousands

Acute effects of TiO2 nanomaterials on the viability and taxonomic composition of aquatic bacterial communities assessed via high-throughput screening and next generation sequencing.

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Binh, Chu Thi Thanh; Tong, Tiezheng; Gaillard, Jean-François; Gray, Kimberly A; Kelly, John J

2014-01-01

The nanotechnology industry is growing rapidly, leading to concerns about the potential ecological consequences of the release of engineered nanomaterials (ENMs) to the environment. One challenge of assessing the ecological risks of ENMs is the incredible diversity of ENMs currently available and the rapid pace at which new ENMs are being developed. High-throughput screening (HTS) is a popular approach to assessing ENM cytotoxicity that offers the opportunity to rapidly test in parallel a wide range of ENMs at multiple concentrations. However, current HTS approaches generally test one cell type at a time, which limits their ability to predict responses of complex microbial communities. In this study toxicity screening via a HTS platform was used in combination with next generation sequencing (NGS) to assess responses of bacterial communities from two aquatic habitats, Lake Michigan (LM) and the Chicago River (CR), to short-term exposure in their native waters to several commercial TiO2 nanomaterials under simulated solar irradiation. Results demonstrate that bacterial communities from LM and CR differed in their sensitivity to nano-TiO2, with the community from CR being more resistant. NGS analysis revealed that the composition of the bacterial communities from LM and CR were significantly altered by exposure to nano-TiO2, including decreases in overall bacterial diversity, decreases in the relative abundance of Actinomycetales, Sphingobacteriales, Limnohabitans, and Flavobacterium, and a significant increase in Limnobacter. These results suggest that the release of nano-TiO2 to the environment has the potential to alter the composition of aquatic bacterial communities, which could have implications for the stability and function of aquatic ecosystems. The novel combination of HTS and NGS described in this study represents a major advance over current methods for assessing ENM ecotoxicity because the relative toxicities of multiple ENMs to thousands of naturally

Florfenicol residues in Rainbow Trout after oral dosing in recirculating and flow-through culture systems

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Meinertz, Jeffery R.; Hess, Karina R.; Bernady, Jeffry A.; Gaikowski, M. P.; Whitsel, Melissa; Endris, R. G.

2014-01-01

Aquaflor is a feed premix for fish containing the broad spectrum antibacterial agent florfenicol (FFC) incorporated at a ratio of 50% (w/w). To enhance the effectiveness of FFC for salmonids infected with certain isolates of Flavobacterium psychrophilum causing coldwater disease, the FFC dose must be increased from the standard 10 mg·kg−1 body weight (BW)·d−1 for 10 consecutive days. A residue depletion study was conducted to determine whether FFC residues remaining in the fillet tissue after treating fish at an increased dose would be safe for human consumption. Groups of Rainbow Trout Oncorhynchus mykiss (total n = 144; weight range, 126–617 g) were treated with FFC at 20 mg·kg−1 BW·d−1 for 10 d in a flow-through system (FTS) and a recirculating aquaculture system (RAS) each with a water temperature of ∼13°C. The two-tank RAS included a nontreated tank containing 77 fish. Fish were taken from each tank (treated tank, n = 16; nontreated tank, n = 8) at 6, 12, 24, 48, 72, 120, 240, 360, and 480 h posttreatment. Florfenicol amine (FFA) concentrations (the FFC marker residue) in skin-on fillets from treated fish were greatest at 12 h posttreatment (11.58 μg/g) in the RAS and were greatest at 6 h posttreatment (11.09 μg/g) in the FTS. The half-lives for FFA in skin-on fillets from the RAS and FTS were 20.3 and 19.7 h, respectively. Assimilation of FFC residues in the fillets of nontreated fish sharing the RAS with FFC-treated fish was minimal. Florfenicol water concentrations peaked in the RAS-treated tank and nontreated tanks at 10 h (453 μg/L) and 11 h (442 μg/L) posttreatment, respectively. Monitoring of nitrite concentrations throughout the study indicated the nitrogen oxidation efficiency of the RAS biofilter was minimally impacted by the FFC treatment.

Culture dependent and independent analysis of bacterial communities associated with commercial salad leaf vegetables

Science.gov (United States)

2013-01-01

Background Plants harbor a diverse bacterial community, both as epiphytes on the plant surface and as endophytes within plant tissue. While some plant-associated bacteria act as plant pathogens or promote plant growth, others may be human pathogens. The aim of the current study was to determine the bacterial community composition of organic and conventionally grown leafy salad vegetables at the point of consumption using both culture-dependent and culture-independent methods. Results Total culturable bacteria on salad vegetables ranged from 8.0 × 103 to 5.5 × 108 CFU g-1. The number of culturable endophytic bacteria from surface sterilized plants was significantly lower, ranging from 2.2 × 103 to 5.8 × 105 CFU g-1. Cultured isolates belonged to six major bacterial phyla, and included representatives of Pseudomonas, Pantoea, Chryseobacterium, and Flavobacterium. Eleven different phyla and subphyla were identified by culture-independent pyrosequencing, with Gammaproteobacteria, Betaproteobacteria, and Bacteroidetes being the most dominant lineages. Other bacterial lineages identified (e.g. Firmicutes, Alphaproteobacteria, Acidobacteria, and Actinobacteria) typically represented less than 1% of sequences obtained. At the genus level, sequences classified as Pseudomonas were identified in all samples and this was often the most prevalent genus. Ralstonia sequences made up a greater portion of the community in surface sterilized than non-surface sterilized samples, indicating that it was largely endophytic, while Acinetobacter sequences appeared to be primarily associated with the leaf surface. Analysis of molecular variance indicated there were no significant differences in bacterial community composition between organic versus conventionally grown, or surface-sterilized versus non-sterilized leaf vegetables. While culture-independent pyrosequencing identified significantly more bacterial taxa, the dominant taxa from pyrosequence data were also detected by

An investigation of the physiology and potential role of components of the deep ocean bacterial community (of the NE Atlantic) by enrichments carried out under minimal environmental change

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Egan, Simon T.; McCarthy, David M.; Patching, John W.; Fleming, Gerard T. A.

2012-03-01

lesser extent members of the Cytophaga-Flavobacterium-Bacteroides and δ groups of prokaryotes.

Sequencing and expression analysis of CD3γ/δ and CD3ɛ chains in mandarin fish, Siniperca chuatsi

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Guo, Zheng; Nie, Pin

2013-01-01

The genomic and cDNA sequences of the CD3γ/δ and CD3ɛ homologues in the mandarin fish, Siniperca chuats i, were determined. As in other vertebrate CD3 molecules, the deduced amino acid sequences of mandarin fish CD3γ/δ and CD3ɛ contained conserved residues and motifs, such as cysteine residues and CXXC and immunoreceptor tyrosine-based activation motifs. However, mandarin fish CD3γ/δ and CD3ɛ showed some differences to their mammalian counterparts, specifically the absence of a negatively charged residue in the transmembrane region of CD3γ/δ. Additionally, while an N -glycosylation site was present in CD3ɛ, the site was not observed in CD3γ/δ. The CD3γ/δ and CD3ɛ subunit sequences contain six and five exons, respectively, consistent with homologues from Atlantic salmon, Salmo salar. Phylogenetic analysis also revealed that CD3γ/δ and CD3ɛ in mandarin fish are closely related to their counterparts in Acanthopterygian fish. Real-time PCR showed CD3γ/δ and CD3ɛ were expressed mainly in the thymus and spleen in normal healthy fish and, to a lesser extent, in mucosal-associated lymphoid tissues, such as the intestine and gills. When lymphocytes isolated from head kidney were treated with the mitogens phytohemagglutinin, concanavalin, and polyriboinosinic polyribocytidylic acid, mRNA expression levels of CD3γ/δ and CD3ɛ were significantly elevated within 12 h of treatment. This indicated the presence of T lymphocytes in the head kidney of teleost fish, and also the recognition of mitogens by the lymphocytes. Mandarin fish infected with the bacterial pathogen Flavobacterium columnare also showed an increase in the expression of CD3γ/δ and CD3ɛ mRNA, indicating that CD3γ/δ and CD3ɛ lymphocytes are involved in the immune response of this species.

Pyrosequencing Reveals Soil Enzyme Activities and Bacterial Communities Impacted by Graphene and Its Oxides.

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Rong, Yan; Wang, Yi; Guan, Yina; Ma, Jiangtao; Cai, Zhiqiang; Yang, Guanghua; Zhao, Xiyue

2017-10-25

Graphene (GN) and graphene oxides (GOs) are novel carbon nanomaterial; they have been attracting much attention because of their excellent properties and are widely applied in many areas, including energy, electronics, biomedicine, environmental science, etc. With industrial production and consumption of GN/GO, they will inevitably enter the soil and water environments. GN/GO may directly cause certain harm to microorganisms and lead to ecological and environmental risks. GOs are GN derivatives with abundant oxygen-containing functional groups in their graphitic backbone. The structure and chemistry of GN show obvious differences compared to those of GO, which lead to the different environmental behaviors. In this study, four different types of soil (S1-S4) were employed to investigate the effect of GN and GO on soil enzymatic activity, microbial population, and bacterial community through pyrosequencing of 16S rRNA gene amplicons. The results showed that soil enzyme activity (invertase, protease, catalase, and urease) and microbial population (bacteria, actinomycetes, and fungi) changed after GN/GO release into soils. Soil microbial community species are more rich, and the diversity also increases after GO/GN application. The phylum of Proteobacteria increased at 90 days after treatment (DAT) after GN/GO application. The phylum of Chloroflexi occurred after GN application at 90 DAT in S1 soil and reached 4.6%. Proteobacteria was the most abundant phylum in S2, S3, and S4 soils; it ranged from 43.6 to 71.4% in S2 soil, from 45.6 to 73.7% in S3 soil, and from 38.1 to 56.7% in S4 soil. The most abundant genera were Bacillus (37.5-47.0%) and Lactococcus (28.0-39.0%) in S1 soil, Lysobacter and Flavobacterium in S2 soil, Pedobacter in S3 soil, and Massilia in S4 soil. The effect of GN and GO on the soil microbial community is time-dependent, and there are no significant differences between the samples at 10 and 90 DAT.

Efficacy of pH elevation as a bactericidal strategy for treating ballast water of freight carriers

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Clifford E. Starliper

2015-05-01

Full Text Available Treatment of ship ballast water with sodium hydroxide (NaOH is one method currently being developed to minimize the risk to introduce aquatic invasive species. The bactericidal capability of sodium hydroxide was determined for 148 bacterial strains from ballast water collected in 2009 and 2010 from the M/V Indiana Harbor, a bulk-freight carrier plying the Laurentian Great Lakes, USA. Primary culture of bacteria was done using brain heart infusion agar and a developmental medium. Strains were characterized based on PCR amplification and sequencing of a portion of the 16S rRNA gene. Sequence similarities (99+ % were determined by comparison with the National Center for Biotechnology Information (NCBI GenBank catalog. Flavobacterium spp. were the most prevalent bacteria characterized in 2009, comprising 51.1% (24/47 of the total, and Pseudomonas spp. (62/101; 61.4% and Brevundimonas spp. (22/101; 21.8% were the predominate bacteria recovered in 2010; together, comprising 83.2% (84/101 of the total. Testing was done in tryptic soy broth (TSB medium adjusted with 5 N NaOH. Growth of each strain was evaluated at pH 10.0, pH 11.0 and pH 12.0, and 4 h up to 72 h. The median cell count at 0 h for 148 cultures was 5.20 × 106 cfu/mL with a range 1.02 × 105–1.60 × 108 cfu/mL. The TSB adjusted to pH 10.0 and incubation for less than 24 h was bactericidal to 52 (35.1% strains. Growth in pH 11.0 TSB for less than 4 h was bactericidal to 131 (88.5% strains and pH 11.0 within 12 h was bactericidal to 141 (95.3%. One strain, Bacillus horikoshii, survived the harshest treatment, pH 12.0 for 72 h.

Efficacy of pH elevation as a bactericidal strategy for treating ballast water of freight carriers.

Science.gov (United States)

Starliper, Clifford E; Watten, Barnaby J; Iwanowicz, Deborah D; Green, Phyllis A; Bassett, Noel L; Adams, Cynthia R

2015-05-01

Treatment of ship ballast water with sodium hydroxide (NaOH) is one method currently being developed to minimize the risk to introduce aquatic invasive species. The bactericidal capability of sodium hydroxide was determined for 148 bacterial strains from ballast water collected in 2009 and 2010 from the M/V Indiana Harbor, a bulk-freight carrier plying the Laurentian Great Lakes, USA. Primary culture of bacteria was done using brain heart infusion agar and a developmental medium. Strains were characterized based on PCR amplification and sequencing of a portion of the 16S rRNA gene. Sequence similarities (99+ %) were determined by comparison with the National Center for Biotechnology Information (NCBI) GenBank catalog. Flavobacterium spp. were the most prevalent bacteria characterized in 2009, comprising 51.1% (24/47) of the total, and Pseudomonas spp. (62/101; 61.4%) and Brevundimonas spp. (22/101; 21.8%) were the predominate bacteria recovered in 2010; together, comprising 83.2% (84/101) of the total. Testing was done in tryptic soy broth (TSB) medium adjusted with 5 N NaOH. Growth of each strain was evaluated at pH 10.0, pH 11.0 and pH 12.0, and 4 h up to 72 h. The median cell count at 0 h for 148 cultures was 5.20 × 10(6) cfu/mL with a range 1.02 × 10(5)-1.60 × 10(8) cfu/mL. The TSB adjusted to pH 10.0 and incubation for less than 24 h was bactericidal to 52 (35.1%) strains. Growth in pH 11.0 TSB for less than 4 h was bactericidal to 131 (88.5%) strains and pH 11.0 within 12 h was bactericidal to 141 (95.3%). One strain, Bacillus horikoshii, survived the harshest treatment, pH 12.0 for 72 h.

The Type IX Secretion System (T9SS: Highlights and Recent Insights into Its Structure and Function

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Anna M. Lasica

2017-05-01

Full Text Available Protein secretion systems are vital for prokaryotic life, as they enable bacteria to acquire nutrients, communicate with other species, defend against biological and chemical agents, and facilitate disease through the delivery of virulence factors. In this review, we will focus on the recently discovered type IX secretion system (T9SS, a complex translocon found only in some species of the Bacteroidetes phylum. T9SS plays two roles, depending on the lifestyle of the bacteria. It provides either a means of movement (called gliding motility for peace-loving environmental bacteria or a weapon for pathogens. The best-studied members of these two groups are Flavobacterium johnsoniae, a commensal microorganism often found in water and soil, and Porphyromonas gingivalis, a human oral pathogen that is a major causative agent of periodontitis. In P. gingivalis and some other periodontopathogens, T9SS translocates proteins, especially virulence factors, across the outer membrane (OM. Proteins destined for secretion bear a conserved C-terminal domain (CTD that directs the cargo to the OM translocon. At least 18 proteins are involved in this still enigmatic process, with some engaged in the post-translational modification of T9SS cargo proteins. Upon translocation across the OM, the CTD is removed by a protease with sortase-like activity and an anionic LPS is attached to the newly formed C-terminus. As a result, a cargo protein could be secreted into the extracellular milieu or covalently attached to the bacterial surface. T9SS is regulated by a two-component system; however, the precise environmental signal that triggers it has not been identified. Exploring unknown systems contributing to bacterial virulence is exciting, as it may eventually lead to new therapeutic strategies. During the past decade, the major components of T9SS were identified, as well as hints suggesting the possible mechanism of action. In addition, the list of characterized cargo

The Type IX Secretion System (T9SS): Highlights and Recent Insights into Its Structure and Function

Science.gov (United States)

Lasica, Anna M.; Ksiazek, Miroslaw; Madej, Mariusz; Potempa, Jan

2017-01-01

Protein secretion systems are vital for prokaryotic life, as they enable bacteria to acquire nutrients, communicate with other species, defend against biological and chemical agents, and facilitate disease through the delivery of virulence factors. In this review, we will focus on the recently discovered type IX secretion system (T9SS), a complex translocon found only in some species of the Bacteroidetes phylum. T9SS plays two roles, depending on the lifestyle of the bacteria. It provides either a means of movement (called gliding motility) for peace-loving environmental bacteria or a weapon for pathogens. The best-studied members of these two groups are Flavobacterium johnsoniae, a commensal microorganism often found in water and soil, and Porphyromonas gingivalis, a human oral pathogen that is a major causative agent of periodontitis. In P. gingivalis and some other periodontopathogens, T9SS translocates proteins, especially virulence factors, across the outer membrane (OM). Proteins destined for secretion bear a conserved C-terminal domain (CTD) that directs the cargo to the OM translocon. At least 18 proteins are involved in this still enigmatic process, with some engaged in the post-translational modification of T9SS cargo proteins. Upon translocation across the OM, the CTD is removed by a protease with sortase-like activity and an anionic LPS is attached to the newly formed C-terminus. As a result, a cargo protein could be secreted into the extracellular milieu or covalently attached to the bacterial surface. T9SS is regulated by a two-component system; however, the precise environmental signal that triggers it has not been identified. Exploring unknown systems contributing to bacterial virulence is exciting, as it may eventually lead to new therapeutic strategies. During the past decade, the major components of T9SS were identified, as well as hints suggesting the possible mechanism of action. In addition, the list of characterized cargo proteins is

The Efficacy of Ultraviolet Radiation for Sterilizing Tools Used for Surgically Implanting Transmitters into Fish

Energy Technology Data Exchange (ETDEWEB)

Walker, Ricardo W.; Markillie, Lye Meng; Colotelo, Alison HA; Gay, Marybeth E.; Woodley, Christa M.; Brown, Richard S.

2013-02-28

Telemetry is frequently used to examine the behavior of fish, and the transmitters used are normally surgically implanted into the coelom of fish. Implantation requires the use of surgical tools such as scalpels, forceps, needle holders, and sutures. When several fish are implanted consecutively for large telemetry studies, it is common for surgical tools to be sterilized or, at minimum, disinfected between each use so that pathogens that may be present are not spread among fish. However, autoclaving tools can take a long period of time, and chemical sterilants or disinfectants can be harmful to both humans and fish and have varied effectiveness. Ultraviolet (UV) radiation is commonly used to disinfect water in aquaculture facilities. However, this technology has not been widely used to sterilize tools for surgical implantation of transmitters in fish. To determine its efficacy for this application, Pacific Northwest National Laboratory researchers used UV radiation to disinfect surgical tools (i.e., forceps, needle holder, stab scalpel, and suture) that were exposed to one of four aquatic organisms that typically lead to negative health issues for salmonids. These organisms included Aeromonas salmonicida, Flavobacterium psychrophilum, Renibacterium salmoninarum, and Saprolegnia parasitica. Surgical tools were exposed to the bacteria by dipping them into a confluent suspension of three varying concentrations (i.e., low, medium, high). After exposure to the bacterial culture, tools were placed into a mobile Millipore UV sterilization apparatus. The tools were then exposed for three different time periods—2, 5, or 15 min. S. parasitica, a water mold, was tested using an agar plate method and forceps-pinch method. UV light exposures of 5 and 15 min were effective at killing all four organisms. UV light was also effective at killing Geobacillus stearothermophilus, the organism used as a biological indicator to verify effectiveness of steam sterilizers. These

Effect of Dursban 480 EC (chlorpyrifos) and Talstar 10 EC (bifenthrin) on the physiological and genetic diversity of microorganisms in soil.

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Medo, Juraj; Maková, Jana; Kovácsová, Silvia; Majerčíková, Kamila; Javoreková, Soňa

2015-01-01

This investigation was undertaken to determine the impact of the insecticides Dursban 480 EC (with organophosphate compound chlorpyrifos as the active ingredient) and Talstar 10 EC (with pyrethroid bifenthrin as the active ingredient) on the respiration activity and microbial diversity in a sandy loam luvisol soil. The insecticides were applied in two doses: the maximum recommended dose for field application (15 mg kg(-1) for Dursban 480 EC and 6 mg kg(-1) for Talstar 10 EC) and a 100-fold higher dose for extrapolation of their effect. Bacterial and fungal genetic diversity was analysed in soil samples using PCR DGGE and the functional diversity (catabolic potential) was studied using BIOLOG EcoPlates at 1, 3, 7, 14, 28, 56 and 112 days after insecticide application. Five bacterial groups (α, β, γ proteobacteria, firmibacteria and actinomycetes) and five groups of fungi or fungus-like microorganisms (Ascomycota, Basidiomycota, Chytridiomycota, Oomycota and Zygomycota) were analysed using specific primer sets. This approach provides high resolution of the analysis covering majority of microorganisms in the soil. Only the high-dose Dursban 480 EC significantly changed the community of microorganisms. We observed its negative effect on α- and γ-proteobacteria, as the number of OTUs (operational taxonomic units) decreased until the end of incubation. In the β-proteobacteria group, initial increase of OTUs was followed by strong decrease. Diversity in the firmibacteria, actinomycetes and Zygomycota groups was minimally disturbed by the insecticide application. Dursban 480 EC, however, both positively and negatively affected certain species. Among negatively affected species Sphingomonas, Flavobacterium or Penicillium were detected, but Achromobacter, Luteibacter or Aspergillus were supported by applied insecticide. The analysis of BIOLOG plates using AWCD values indicated a significant increase in metabolic potential of microorganisms in the soil after the high

Characterization of biofilm in 200W fluidized bed reactors

Energy Technology Data Exchange (ETDEWEB)

Lee, Michelle H. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Saurey, Sabrina D. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Lee, Brady D. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Parker, Kent E. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Eisenhauer, Emalee E. R. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Cordova, Elsa A. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Golovich, Elizabeth C. [Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

2014-09-29

Contaminated groundwater beneath the 200 West Area at the Hanford Site in Southeast Washington is currently being treated using a pump and treat system to remove organics, inorganics, radionuclides, and metals. A granular activated carbon-based fluidized bed reactor (FBR) has been added to remove nitrate, hexavalent chromium and carbon tetrachloride. Initial analytical results indicated the microorganisms effectively reduced many of the contaminants to less than cleanup levels. However shortly thereafter operational upsets of the FBR include carbon carry over, over production of microbial extracellular polymeric substance (biofilm) materials, and over production of hydrogen sulfide. As a result detailed investigations were undertaken to understand the functional diversity and activity of the microbial community present in the FBR over time. Molecular analyses including terminal restriction fragment length polymorphism analysis, quantitative polymerase chain reaction and fluorescent in situ hybridization analyses were performed on the microbial community extracted from the biofilm within the bed and from the inoculum, to determine functional dynamics of the FBR bed over time and following operational changes. Findings from these analyses indicated: 1) the microbial community within the bed was completely different than community used for inoculation, and was likely from the groundwater; 2) analyses early in the testing showed an FBR community dominated by a few Curvibacter and Flavobacterium species; 3) the final sample taken indicated that the microbial community in the FBR bed had become more diverse; and 4) qPCR analyses indicated that bacteria involved in nitrogen cycling, including denitrifiers and anaerobic ammonia oxidizing bacteria, were dominant in the bed. These results indicate that molecular tools can be powerful for determining functional diversity within FBR type reactors. Coupled with micronutrient, influent and effluent chemistry

Metatranscriptome Sequencing Reveals Insights into the Gene Expression and Functional Potential of Rumen Wall Bacteria

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Evelyne Mann

2018-01-01

Full Text Available Microbiota of the rumen wall constitute an important niche of rumen microbial ecology and their composition has been elucidated in different ruminants during the last years. However, the knowledge about the function of rumen wall microbes is still limited. Rumen wall biopsies were taken from three fistulated dairy cows under a standard forage-based diet and after 4 weeks of high concentrate feeding inducing a subacute rumen acidosis (SARA. Extracted RNA was used for metatranscriptome sequencing using Illumina HiSeq sequencing technology. The gene expression of the rumen wall microbial community was analyzed by mapping 35 million sequences against the Kyoto Encyclopedia for Genes and Genomes (KEGG database and determining differentially expressed genes. A total of 1,607 functional features were assigned with high expression of genes involved in central metabolism, galactose, starch and sucrose metabolism. The glycogen phosphorylase (EC:2.4.1.1 which degrades (1->4-alpha-D-glucans was among the highest expressed genes being transcribed by 115 bacterial genera. Energy metabolism genes were also highly expressed, including the pyruvate orthophosphate dikinase (EC:2.7.9.1 involved in pyruvate metabolism, which was covered by 177 genera. Nitrogen metabolism genes, in particular glutamate dehydrogenase (EC:1.4.1.4, glutamine synthetase (EC:6.3.1.2 and glutamate synthase (EC:1.4.1.13, EC:1.4.1.14 were also found to be highly expressed and prove rumen wall microbiota to be actively involved in providing host-relevant metabolites for exchange across the rumen wall. In addition, we found all four urease subunits (EC:3.5.1.5 transcribed by members of the genera Flavobacterium, Corynebacterium, Helicobacter, Clostridium, and Bacillus, and the dissimilatory sulfate reductase (EC 1.8.99.5 dsrABC, which is responsible for the reduction of sulfite to sulfide. We also provide in situ evidence for cellulose and cellobiose degradation, a key step in fiber-rich feed

Influence of chosen microbes and some chemical substances on the production of aflatoxins

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Iveta Brožková

2015-03-01

Full Text Available Aflatoxins are produced as secondary metabolites by A. flavus, A. parasiticus, A. nomius and A. tamarii. The aflatoxin biosynthetic pathway involves several enzymatic steps and genes (apa-2, ver-1 that appear to be regulated by the aflR gene in these fungi. The aim of this work was the detection of aflatoxins by the HPLC method and the ascertainment of factors influencing their production. A. parasiticus CCM F-108, A. parasiticus CCF 141, A. parasiticus CCF 3137 and two isolates A. flavus were used. These toxigenic isolates were recovered from spice (strain 1 and wraps (strain 2. The gene for the production of aflatoxin B1 for each species of fungi was detected using an optimized PCR method. Rhodotorula spp.*, Lactococcus lactis subsp. lactis CCM 1881, Flavobacterium spp. and fungal strain Pythium oligandrum* were tested for inhibition of aflatoxins production and fungal growth. Having used the HPLC detection, various preservatives (propionic acid, citric acid, potassium sorbate were tested from the viewpoint of their influence on the growth of aflatoxigenic fungi followed by the production of aflatoxins. The growth of A. flavus and A. parasiticus and aflatoxin production in Potato Dextrose Agar supplemented with propionic acid (1000-2000-3000 mg/kg, citric acid (2000-3000-4000 mg/kg and potassium sorbate (500-800-1000 mg/kg was tested by Agar Dilution Method. After 72 h of incubation was evaluated growth of fungi, all samples were frozen for later extraction and aflatoxins quantification by HPLC. Effect of peptone and sucrose additions were studied in yeast extract (2% supplemented with peptone (5-10-15% or sucrose (15%. Growth inhibition of Aspergillus by Pythium oligandrum was tested on wood surface. As shown, the highest inhibition effect on the aflatoxins production was obtained when propionic acid was applied in concentrations since 1000 mg/kg. A total inhibition of the fungi growth and aflatoxins production was observed in all samples

Ultraviolet radiation as disinfection for fish surgical tools

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Walker, Ricardo W.; Markillie, Lye Meng; Colotelo, Alison HA; Geist, David R.; Gay, Marybeth E.; Woodley, Christa M.; Eppard, M. B.; Brown, Richard S.

2013-04-04

Telemetry is frequently used to examine the behavior of fish, and the transmitters used are normally surgically implanted into the coelomic cavity of fish. Implantation requires the use of surgical tools such as scalpels, forceps, needle holders, and sutures. When fish are implanted consecutively, as in large telemetry studies, it is common for surgical tools to be sterilized or, at minimum, disinfected between each use so that pathogens that may be present are not spread among fish. To determine the efficacy for this application, ultraviolet (UV) radiation was used to disinfect surgical tools exposed to one of four aquatic organisms that typically lead to negative health issues for salmonids. These organisms included Aeromonas salmonicida, Flavobacterium psychrophilum, Renibacterium salmoninarum, and Saprolegnia parasitica, causative agents of furunculosis, coldwater disease, bacterial kidney disease, and saprolegniasis (water mold), respectively. Four experiments were conducted to address the question of UV efficacy. In the first experiment, forceps were exposed to the three bacteria at three varying concentrations. After exposure to the bacterial culture, tools were placed into a mobile Millipore UV sterilization apparatus. The tools were then exposed for three different time periods – 2, 5, or 15 min. UV radiation exposures at all durations were effective at killing all three bacteria on forceps at the highest bacteria concentrations. In the second experiment, stab scalpels, sutures, and needle holders were exposed to A. salmonicida using the same methodology as used in Experiment 1. UV radiation exposure at 5 and 15 min was effective at killing A. salmonicida on stab scalpels and sutures but not needle holders. In the third experiment, S. parasitica, a water mold, was tested using an agar plate method and forceps-pinch method. UV radiation was effective at killing the water mold at all three exposure durations. Collectively, this study shows that UV

Half-lives of PAHs and temporal microbiota changes in commonly used urban landscaping materials

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Marja I. Roslund

2018-03-01

Full Text Available Background Polycyclic aromatic hydrocarbons (PAHs accumulate in urban soils, and PAH contamination can change soil microbial community composition. Environmental microbiota is associated with human commensal microbiota, immune system and health. Therefore, studies investigating the degradation of PAHs, and the consequences of soil pollution on microbial communities in urban landscaping materials, are crucial. Methods Four landscaping materials (organic matter 1, 2, 13 and 56% were contaminated with PAHs commonly found at urban sites (phenanthrene, fluoranthene, pyrene, chrysene and benzo(bfluoranthene in PAH concentrations that reflect urban soils in Finland (2.4 µg g -1 soil dry weight. PAHs were analyzed initially and after 2, 4, 8 and 12 weeks by gas chromatography-mass spectrometry. Half-lives of PAHs were determined based on 12-weeks degradation. Bacterial communities were analyzed at 1 and 12 weeks after contamination using Illumina MiSeq 16S rRNA gene metabarcoding. Results Half-lives ranged from 1.5 to 4.4 weeks for PAHs with relatively low molecular weights (phenanthrene, fluoranthene and pyrene in landscaping materials containing 1–2% organic matter. In contrast, in materials containing 13% and 56% organic matter, the half-lives ranged from 2.5 to 52 weeks. Shorter half-lives of phenanthrene and fluoranthene were thus associated with low organic matter content. The half-life of pyrene was inversely related to the relative abundance of Beta-, Delta- and Gammaproteobacteria, and diversity of Bacteroidetes and Betaprotebacteria. Compounds with higher molecular weights followed compound-specific patterns. Benzo(bfluoranthene was resistant to degradation and half-life of chrysene was shorter when the relative abundance of Betaproteobacteria was high. Temporal microbiota changes involved increase in the relative abundance of Deltaproteobacteria and decrease in genera Flavobacterium and Rhodanobacter. Exposure to PAHs seems to adjust

Corexit 9500 Enhances Oil Biodegradation and Changes Active Bacterial Community Structure of Oil-Enriched Microcosms.

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Techtmann, Stephen M; Zhuang, Mobing; Campo, Pablo; Holder, Edith; Elk, Michael; Hazen, Terry C; Conmy, Robyn; Santo Domingo, Jorge W

2017-05-15

To better understand the impacts of Corexit 9500 on the structure and activity levels of hydrocarbon-degrading microbial communities, we analyzed next-generation 16S rRNA gene sequencing libraries of hydrocarbon enrichments grown at 5 and 25°C using both DNA and RNA extracts as the sequencing templates. Oil biodegradation patterns in both 5 and 25°C enrichments were consistent with those reported in the literature (i.e., aliphatics were degraded faster than aromatics). Slight increases in biodegradation were observed in the presence of Corexit at both temperatures. Differences in community structure were observed between treatment conditions in the DNA-based libraries. The 25°C consortia were dominated by Vibrio , Idiomarina , Marinobacter , Alcanivorax , and Thalassospira species, while the 5°C consortia were dominated by several species of the genera Flavobacterium , Alcanivorax , and Oleispira Most of these genera have been linked to hydrocarbon degradation and have been observed after oil spills. Colwellia and Cycloclasticus , known aromatic degraders, were also found in these enrichments. The addition of Corexit did not have an effect on the active bacterial community structure of the 5°C consortia, while at 25°C, a decrease in the relative abundance of Marinobacter was observed. At 25°C, Thalassospira , Marinobacter , and Idiomarina were present at higher relative abundances in the RNA than DNA libraries, suggesting that they were active in degradation. Similarly, Oleispira was greatly stimulated by the addition of oil at 5°C. IMPORTANCE While dispersants such as Corexit 9500 can be used to treat oil spills, there is still debate on the effectiveness on enhancing oil biodegradation and its potential toxic effect on oil-degrading microbial communities. The results of this study provide some insights on the microbial dynamics of hydrocarbon-degrading bacterial populations in the presence of Corexit 9500. Operational taxonomic unit (OTU) analyses

Changes in {sup 137}Cs bioavailability under the influence of soil microflora

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Pareniuk, Olena [Institute of Environmental Radioactivity of Fukushima University, 1 Kanayagawa, Fukushima City, Fukushima Prefecture, 960-1296 (Japan); Ukrainian Institute of Agricultural Radiology of National University of Life and Environmental Sciences of Ukraine, Chabany, Kyivo-Svyatoshin region, 08162, Kyiv (Ukraine); Illienko, Volodymyr; Gudkov, Igor [National University of Life and Environmental Sciences of Ukraine, Heroyiv Oborony st., 15, Kyiv-03041 (Ukraine); Shavanova, Kateryna; Levchuk, Svyatoslav [Ukrainian Institute of Agricultural Radiology of National University of Life and Environmental Sciences of Ukraine, Chabany, Kyivo-Svyatoshin region, 08162, Kyiv (Ukraine)

2014-07-01

Despite the rising safety standards applicable to the nuclear industry enterprises still occur the huge radiation accidents, the consequences of which are large-scale releases of radionuclides into the environment. The soil is the main depot for radioactive substances in the ecosystem and triggering element for their transition through trophic chains up to the human being. That is why the search for new effective methods of soil decontamination, particularly the soil that is used for agriculture, still remains of a big relevance. Using the properties of microorganisms to affect the availability of mineral compounds in the soil may become one of such methods. {sup 137}Cs is one of the main dose-forming radionuclides, introduced into the ecosystem as a result of the nuclear weapons testing and accidents occurring at nuclear power plants. Thus, radionuclide contamination of agricultural land and food is a major problem for the exclusion zones, formed after the accidents at the Fukushima Daiichi and the Chernobyl NPPs. To study the role of soil microorganisms in the migration of {sup 137}Cs 5 isolates from Chernobyl exclusion zone were allocated and identified in accordance with 16s RNA: Burkholderia glathei Hg11, Burkholderia sp IMER-B1-53, Bacillus mycides BCHMAC12, Flavobacterium sp TISTR 1602, Pseudomonas frederiksbergensis TPK 2-4 and compared their ability to accumulate radionuclide with collection species, which are used as a bio-agents of microbial fertilizers: Bacillusmegaterium UKM B-5724, Azotobacterchroococcum UKM B-6003-20A, Azotobacterchroococcum UKM B-6003-9T, Agrobacterium radiobacter by transfer of isolates to liquid nutrient medium containing 5 kBq {sup 137}Cs. It was found that the strain Bacillusmegaterium UKM B-5724 from the collection of the Institute of Microbiology and Virology of NASU, has a high ability to accumulate radionuclides. Figured out that certain types of microorganisms can either reduce or increase the ratio of {sup 137}Cs transfer

Use of Endophytic and Rhizosphere Bacteria To Improve Phytoremediation of Arsenic-Contaminated Industrial Soils by Autochthonous Betula celtiberica.

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Mesa, Victoria; Navazas, Alejandro; González-Gil, Ricardo; González, Aida; Weyens, Nele; Lauga, Béatrice; Gallego, Jose Luis R; Sánchez, Jesús; Peláez, Ana Isabel

2017-04-15

The aim of this study was to investigate the potential of indigenous arsenic-tolerant bacteria to enhance arsenic phytoremediation by the autochthonous pseudometallophyte Betula celtiberica The first goal was to perform an initial analysis of the entire rhizosphere and endophytic bacterial communities of the above-named accumulator plant, including the cultivable bacterial species. B. celtiberica 's microbiome was dominated by taxa related to Flavobacteriales , Burkholderiales , and Pseudomonadales , especially the Pseudomonas and Flavobacterium genera. A total of 54 cultivable rhizobacteria and 41 root endophytes, mainly affiliated with the phyla Proteobacteria , Bacteroidetes , Firmicutes , and Actinobacteria , were isolated and characterized with respect to several potentially useful features for metal plant accumulation, such as the ability to promote plant growth, metal chelation, and/or mitigation of heavy-metal stress. Seven bacterial isolates were further selected and tested for in vitro accumulation of arsenic in plants; four of them were finally assayed in field-scale bioaugmentation experiments. The exposure to arsenic in vitro caused an increase in the total nonprotein thiol compound content in roots, suggesting a detoxification mechanism through phytochelatin complexation. In the contaminated field, the siderophore and indole-3-acetic acid producers of the endophytic bacterial consortium enhanced arsenic accumulation in the leaves and roots of Betula celtiberica , whereas the rhizosphere isolate Ensifer adhaerens strain 91R mainly promoted plant growth. Field experimentation showed that additional factors, such as soil arsenic content and pH, influenced arsenic uptake in the plant, attesting to the relevance of field conditions in the success of phytoextraction strategies. IMPORTANCE Microorganisms and plants have developed several ways of dealing with arsenic, allowing them to resist and metabolize this metalloid. These properties form the basis of

Community composition and cellulase activity of cellulolytic bacteria from forest soils planted with broad-leaved deciduous and evergreen trees.

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Yang, Jiang-Ke; Zhang, Jing-Jing; Yu, Heng-Yu; Cheng, Jian-Wen; Miao, Li-Hong

2014-02-01

Cellulolytic bacteria in forest soil provide carbon sources to improve the soil fertility and sustain the nutrient balance of the forest ecological system through the decomposition of cellulosic remains. These bacteria can also be utilized for the biological conversion of biomass into renewable biofuels. In this study, the community compositions and activities of cellulolytic bacteria in the soils of forests planted with broad-leaved deciduous (Chang Qing Garden, CQG) and broad-leaved evergreen (Forest Park, FP) trees in Wuhan, China were resolved through restriction fragment length polymorphism (RFLP) and sequencing analysis of the 16S rRNA gene. All of the isolates exhibited 35 RFLP fingerprint patterns and were clustered into six groups at a similarity level of 50 %. The phylogeny analysis based on the 16S rRNA gene sequence revealed that these RFLP groups could be clustered into three phylogenetic groups and further divided into six subgroups at a higher resolution. Group I consists of isolates from Bacillus cereus, Bacillus subtilis complex (I-A) and from Paenibacillus amylolyticus-related complex (I-B) and exhibited the highest cellulase activity among all of the cellulolytic bacteria isolates. Cluster II consists of isolates belonging to Microbacterium testaceum (II-A), Chryseobacterium indoltheticum (II-B), and Flavobacterium pectinovorum and the related complex (II-C). Cluster III consists of isolates belonging to Pseudomonas putida-related species. The community shift with respect to the plant species and the soil properties was evidenced by the phylogenetic composition of the communities. Groups I-A and I-B, which account for 36.0 % of the cellulolytic communities in the CQG site, are the dominant groups (88.4 %) in the FP site. Alternatively, the ratio of the bacteria belonging to group III (P. putida-related isolates) shifted from 28.0 % in CQG to 4.0 % in FP. The soil nutrient analysis revealed that the CQG site planted with deciduous broad

Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast

Energy Technology Data Exchange (ETDEWEB)

Eudes, Aymerick [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Mouille, Maxence [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Robinson, David S. [Joint BioEnergy Institute, Emeryville, CA (United States); Benites, Veronica T. [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); San Francisco State Univ., San Francisco, CA (United States); Wang, George [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Roux, Lucien [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Ecole Polytechnique Federale de Lausanne, Lausanne (Switzerland); Tsai, Yi-Lin [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Baidoo, Edward E. K. [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Chiu, Tsan-Yu [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Heazlewood, Joshua L. [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); The Univ. of Melbourne, Melbourne, VIC (Australia); Scheller, Henrik V. [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Mukhopadhyay, Aindrila [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Keasling, Jay D. [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States); Technical Univ. of Denmark, Horsholm (Denmark); Deutsch, Samuel [Joint BioEnergy Institute, Emeryville, CA (United States); Loqué, Dominique [Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. Claude Bernard Lyon 1, Villeurbanne (France)

2016-11-21

. Similarly, we achieved for the first time the microbial production of polyamine hydroxycinnamate amides; monolignol, malate and fatty alcohol hydroxycinnamate esters; tropane alkaloids; and benzoate/caffeate alcohol esters. In some instances, the additional expression of Flavobacterium johnsoniae tyrosine ammonia-lyase (FjTAL) allowed the synthesis of p-coumarate conjugates and eliminated the need to supplement the culture media with 4-hydroxycinnamate. In conclusion, we demonstrate in this study the effectiveness of expressing members of the plant BAHD acyltransferase family in yeast for the synthesis of numerous valuable hydroxycinnamate and benzoate conjugates.

Lacinutrix cladophorae sp. nov., a flavobacterium isolated from the green alga Cladophora stimpsonii, transfer of Flavirhabdus iliipiscaria Shakeela et al. 2015 to the genus Lacinutrix as Lacinutrix iliipiscaria comb. nov. and emended description of the genus Lacinutrix.

Science.gov (United States)

Nedashkovskaya, Olga I; Kim, Song-Gun; Zhukova, Natalia V; Lee, Jung-Sook; Mikhailov, Valery V

2016-11-01

A strictly aerobic, Gram-stain-negative, rod-shaped, motile by gliding and yellow-pigmented bacterium, designated strain 7Alg 4T, was isolated from the green alga Cladophora stimpsonii. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the novel strain was affiliated to the family Flavobacteriaceae of the phylum Bacteroidetes, and was most closely related to the recognized species of the genera Lacinutrixand Flavirhabdus, with 16S rRNA gene sequence similarities of 95.1-98.1 and 97.0 %, respectively. Strain 7Alg 4T grew in the presence of 1-5 % NaCl and at 4-32 °C, and hydrolysed aesculin, gelatin, starch and Tween 80. The prevalent fatty acids were iso-C15 : 1 G, iso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 0 3-OH and C15 : 0. The polar lipid profile was characterized by the presence of phosphatidylethanolamine, three unidentified aminolipids and four unidentified lipids. The major respiratory quinone was MK-6. The DNA G+C content was 31.9 mol%. On the basis of the differences in 16S rRNA gene sequences, chemotaxonomic and phenotypic characteristics, it is suggested that strain 7Alg 4T represents a novel species of the genus Lacinutrix, for which the name Lacinutrixcladophorae sp. nov. is proposed. The type strain is 7Alg 4T (=KCTC 23036T=KMM 6381T). Reclassification of Flavirhabdus iliipiscaria as Lacinutrix iliipiscaria comb. nov. and an emend of the genus Lacinutrix are also proposed.

Microbiological Aspects of Geothermal Energy: Influence of Microbial Activity on Scaling and Clogging in a Cold Storage

Science.gov (United States)

Lerm, Stephanie; Alawi, Mashal; Miethling-Graff, Rona; Vieth, Andrea; Seibt, Andrea; Wolfgramm, Markus; Würdemann, Hilke

2010-05-01

that bacteria and their metabolic activities were involved in the decrease of filter endurances. A strong biofilm formation of filamentous sulfur-oxidizing bacteria related to Thiothrix was observed. In the course of the disinfection measure the microbial composition in the process water changed significantly. Thiothrix could not be detected any longer and the biocoenosis in the fluid was dominated now by Flavobacterium, Acidovorax as well as Alcaligenaceae related organisms. In contrast, SRB analyzed by specific dissimilatory sulfite reductase genes were hardly affected by the disinfection measures. However, even if especially SRB are considered as the most important taxonomic group for microbiologically influenced corrosion (MIC), present operational results indicate that scaling and clogging were the predominant processes for the operation of the shallow cold storage in Berlin.

Research on the impact of polyhexamethyleneguanidine on the plant component of biocenoses

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A. V. Lysytsya

2017-05-01

Full Text Available This article analyses the results of studying the influence of polymeric guanidine derivatives, namely polyhexa­methyleneguanidine chloride (PНMG, on land and freshwater plants. Gas chromatography was used to determine the soil – plant and water – plant transition coefficients. Methods of mass spectrometry and photocalorimetry were used to identify the PHMG in the samples. We investigated the toxicity of PНMG for freshwater flora in an aquarium on Vallisneria spiralis, Riccia fluitans and Chlorella pyrenoidosa. The results showed that even the lowest bactericidal concentrations of the preparation (10–3% or 10 mg/L caused the death of test organisms within one to two days. One-time application of PНMG to the aquarium in dose of 10–4% (or 1 mg/L did not cause any noticeable changes in algae during the 7 days of the experiment. The PНMG transfer coefficient did not exceed 0.1% for the system "water – algae tissue". Moreover, the initial concentration of the drug in the water decreased by almost ten times already during the first two days. PНMG polycation molecules quickly bind to dissolved in water organic and inorganic substances, suspended particles, microorganisms, etc. Flocculation causes a sharp decrease in the number of active "free" polycation molecules in water. The drug settles on the bottom of the aquarium and then is destroyed by bacteria saprophytes. Apparently, PHMG is included in their metabolism and serves as nitrogen source for microorganisms Pseudomonas putida, Flavobacterium columnare, Bacillus sp., Sarcina sp., Nitrosomonas sp. and Nitrobacter sp. At the same time, this study showed that the safe concentration of PHMG for hydrobionts in the water of natural fresh water reservoirs is 0.01 mg/l, or 10–6%, provided the drug is chronic. Ground plants are more resistant to the action of PHMG. They easiliy tolerate finely dispersed spraying with 0.3% aqueous solution of PGMG chloride in a dose of 0.5–1.0 l/m2. For

Identity of zinc finger nucleases with specificity to herpes simplex virus type II genomic DNA: novel HSV-2 vaccine/therapy precursors

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Wayengera Misaki

2011-06-01

Full Text Available Abstract Background Herpes simplex type II (HSV-2 is a member of the family herpesviridae. Human infection with this double stranded linear DNA virus causes genital ulcerative disease and existing treatment options only serve to resolve the symptomatology (ulcers associated with active HSV-2 infection but do not eliminate latent virus. As a result, infection with HSV-2 follows a life-long relapsing (active versus latent course. On the basis of a primitive bacterium anti-phage DNA defense, the restriction modification (R-M system, we previously identified the Escherichia coli restriction enzyme (REase EcoRII as a novel peptide to excise or irreversibly disrupt latent HSV-2 DNA from infected cells. However, sequences of the site specificity palindrome of EcoRII 5'-CCWGG-3' (W = A or T are equally present within the human genome and are a potential source of host-genome toxicity. This feature has limited previous HSV-2 EcoRII based therapeutic models to microbicides only, and highlights the need to engineer artificial REases (zinc finger nucleases-ZFNs with specificity to HSV-2 genomic-DNA only. Herein, the therapeutic-potential of zinc finger arrays (ZFAs and ZFNs is identified and modeled, with unique specificity to the HSV-2 genome. Methods and results Using the whole genome of HSV-2 strain HG52 (Dolan A et al.,, and with the ZFN-consortium's CoDA-ZiFiT software pre-set at default, more than 28,000 ZFAs with specificity to HSV-2 DNA were identified. Using computational assembly (through in-silico linkage to the Flavobacterium okeanokoites endonuclease Fok I of the type IIS class, 684 ZFNs with specificity to the HSV-2 genome, were constructed. Graphic-analysis of the HSV-2 genome-cleavage pattern using the afore-identified ZFNs revealed that the highest cleavage-incidence occurred within the 30,950 base-pairs (~between the genomic context coordinates 0.80 and 1.00 at the 3' end of the HSV-2 genome. At approximately 3,095 bp before and after the

[Phylogenetic diversity and cold-adaptive hydrolytic enzymes of culturable psychrophilic bacteria associated with sea ice from high latitude ocean, Artic].

Science.gov (United States)

Yu, Yong; Li, Hui-Rong; Chen, Bo; Zeng, Yin-Xin; He, Jian-Feng

2006-04-01

The phylogenetic diversity of culturable psychrophilic bacteria associated with sea ice from high latitude sea (77 degrees 30'N - 81 degrees 12'N), Canadian Basin and Greenland sea Arctic, was investigated. A total of 37 psychrophilic strains were isolated using three different methods of ( i ) spread plate method: 100 microL of each dilution ice-melt sample was spreaded onto the surface of Marine 2216 agar (DIFCO laboratories, Detroit, MI) and incubated for 2 to 6 weeks at 4 degrees C; ( ii ) bath culture and spread plate method: 1 mL of sample was added to 9mL of NSW (unamended natural seawater, 0.2 microm prefiltered and autoclaved) and incubated for 1 months at - 1 degrees C, then spread plate method was used to isolate bacterial strains from the pre-cultured samples; ( iii ) cold shock, bath culture and spread plate method: samples were exposed to - 20 degrees C for 24h, then bacterial strains isolated by bath culture and spread plate method under aerobic conditions. Nearly half of psychrophilic strains are isolated by using method iii . 16S rDNA nearly full-length sequence analysis reveal that psychrophilic strains fall in two phylogenetic divisions, gamma-proteobacteria (in the genera Colwellia, Marinobacter, Shewanella, Thalassomonas, Glaciecola, Marinomonas and Pseudoalteromonas) and Cytophaga-Flexibacter-Bacteroides (in the genera Flavobacterium and Psychroflexus). Nine of bacterial isolates (BSi20007, BSi20497, BSi20517, BSi20537, BSi20170, BSi20001, BSi20002, BSi20675 and BSi20101) quite likely represent novel species (16S rDNA sequence similarity below 97%). One of strains (BSi20002) from Canadian Basin shows 100% sequence similarity to the Antarctic Weddell sea ice isolate Marinobacter sp. ANT8277, suggesting bacteria may have a bipolar distribution at the species level. AF283859 sequences were submitted to the BLAST search program of the National Center for Biotechnology Information website (NCBI, http://www. ncbi. nlm.nih. gov). Twenty sequences

Escapement and Productivity of Spring Chinook and Summer Steelhead in the John Day River Basin, Technical Report 2004-2005.

Energy Technology Data Exchange (ETDEWEB)

Wilson, Wayne

2007-04-01

The objectives are: (1) Estimate number and distribution of spring Chinook salmon Oncorhynchus tshawytscha redds and spawners in the John Day River subbasin; and (2) Estimate smolt-to-adult survival rates (SAR) and out-migrant abundance for spring Chinook and summer steelhead O. mykiss and life history characteristics of summer steelhead. Spawning ground surveys for spring (stream-type) Chinook salmon were conducted in four main spawning areas (Mainstem, Middle Fork, North Fork, and Granite Creek System) and seven minor spawning areas (South Fork, Camas Creek, Desolation Creek, Trail Creek, Deardorff Creek, Clear Creek, and Big Creek) in the John Day River basin during August and September of 2005. Census surveys included 298.2 river kilometers (88.2 rkm within index, 192.4 rkm additional within census, and 17.6 rkm within random survey areas) of spawning habitat. We observed 902 redds and 701 carcasses including 227 redds in the Mainstem, 178 redds in the Middle Fork, 420 redds in the North Fork, 62 redds in the Granite Creek System, and 15 redds in Desolation Creek. Age composition of carcasses sampled for the entire basin was 1.6% age 3, 91.2% age 4, and 7.1% age 5. The sex ratio was 57.4% female and 42.6% male. Significantly more females than males were observed in the Granite Creek System. During 2005, 82.3% of female carcasses sampled had released all of their eggs. Significantly more pre-spawn mortalities were observed in Granite Creek. Nine (1.3%) of 701 carcasses were of hatchery origin. Of 298 carcasses examined, 4.0% were positive for the presence of lesions. A significantly higher incidence of gill lesions was found in the Granite Creek System when compared to the rest of the basin. Of 114 kidney samples tested, two (1.8%) had clinical BKD levels. Both infected fish were age-4 females in the Middle Fork. All samples tested for IHNV were negative. To estimate spring Chinook and summer steelhead smolt-to-adult survival (SAR) we PIT tagged 5,138 juvenile

Characterization of Microbial Communities Associated With Deep-Sea Hydrothermal Vent Animals of the East Pacific Rise and the Galápagos Rift

Science.gov (United States)

Ward, N.; Page, S.; Heidelberg, J.; Eisen, J. A.; Fraser, C. M.

2002-12-01

epsilon proteobacteria, low GC Gram positive bacteria (firmicutes), spirochetes, CFB (Cytophaga-Flavobacterium-Bacteroides) group, green nonsulfur bacteria, acidobacteria, verrucomicrobia, and planctomycetes. The presence of the latter three taxonomic groups is of special interest, as they represent phylogenetically distinct groups within the Bacteria for which specific ecological functions have not yet been identified, but which have been found to be widely distributed and often numerically significant in diverse terrestrial and aquatic habitats. Although further sequencing is required to demonstrate the presence of a Riftia-associated microbial population distinct from that of the surrounding seawater, results available from three Riftia individuals from the East Pacific Rise suggest this to be the case. Analysis of microbial communities associated with the gill tissue of the mussel Bathymodiolus thermophilus shows a population dominated by gamma-Proteobacterial chemoautotrophic symbionts, although lower frequency novel phylotypes have been detected. Representatives of specific taxonomic groups have been selected for sequencing of the complete 16S rRNA gene, and the sequences used to reconstruct phylogenetic trees to more accurately determine the evolutionary relationships between the novel sequences, and available sequences for both cultured and non-cultured bacteria.

Minthorn Springs Creek summer juvenile release and adult collection facility: Annual report 1992; ANNUAL

International Nuclear Information System (INIS)

Rowan, Gerald D.

1993-01-01

The Confederated Tribes of the Umatilla Indian Reservation (CT'UIR) and Oregon Department of Fish and Wildlife (ODFW) are cooperating in a joint effort to supplement steelhead and re-establish salmon runs in the Umatilla River Basin. As an integral part of this program, Bonifer and Minthorn Acclimation Facilities are operated for holding and spawning adult steelhead and fall chinook salmon and acclimation and release of juvenile salmon and steelhead. Acclimation of 109,101 spring chinook salmon and 19,977 summer steelhead was completed at Bonifer in the spring of 1992. At Minthorn, 47,458 summer steelhead were acclimated and released. Control groups of spring chinook salmon were released instream concurrent with the acclimated releases to evaluate the effects of acclimation on adult returns to the Umatilla River. Acclimation studies with summer steelhead were not conducted in 1992. A total of 237 unmarked adult steelhead were collected for broodstock at Three Mile Dam from October 18, 1991 through April 24, 1992 and held at Minthorn. Utilizing a 3 x 3 spawning matrix, a total of 476,871 green eggs were taken from 86 females. The eggs were transferred to Umatilla Hatchery for incubation, rearing, and later release into the Umatilla River. A total of 211 fall chinook salmon were also collected for broodstock at Three Mile Dam and held at Minthorn. Using a 1:1 spawning ratio, a total of 195,637 green eggs were taken from 58 females. They were also transferred to Umatilla Hatchery for incubation, rearing, and later release into the Umatilla River. Personnel from the ODFW Eastern Oregon Fish Pathology Laboratory in La Grande took samples of tissues and reproductive fluids from Umatilla River summer steelhead and fall chinook salmon broodstock for monitoring and evaluation purposes. Cell culture assays for replicating agents, including IHNV virus, on all spawned fish were negative. One of 60 summer steelhead tested positive for EIBS virus, while all fall chinook tested

Marine echinoderms as reservoirs of antimicrobial resistant bacteria

Directory of Open Access Journals (Sweden)

Catarina Marinho

2014-06-01

Full Text Available Echinoderms are benthic animals that play an important ecological role in marine communities occupying diverse trophic levels in the marine food chains. The majority of echinoderms feed on small particles of edible matter, although they can eat many kinds of food (Clark, 1968. Although, some echinoderms species has been facing an emerging demand for human consumption, particularly in Asian and Mediterranean cuisine, where these animals can be eaten raw (Kelly, 2005; Micael et al., 2009. Echinoderms own an innate immune mechanism that allows them to defend themselves from high concentrations of bacteria, viruses and fungus they are often exposed, on marine sediment (Janeway and Medzhitov, 1998, Cooper, 2003. The most frequent genera of gut bacteria in echinoderms are Vibrio, Pseudomonas, Flavobacterium, and Aeromonas; nevertheless Enterococcus spp. and Escherichia coli are also present (Harris, 1993; Marinho et al., 2013. Moreover, fecal resistant bacteria found in the aquatic environment might represent an index of marine pollution (Foti et al., 2009, Kummerer, 2009. Several studies had been lead in order to identify environmental reservoirs for antibiotic-resistant bacteria in populations of fish, echinoderms and marine mammals, and they all support the thesis that these animals may serve as reservoirs since they had acquired resistant microbial species (Johnson et al., 1998, Marinho et al., 2013, Miranda and Zemelman, 2001. However, to our knowledge, there are only available in bibliography one study of antimicrobial resistant bacteria isolated from marine echinoderms (Marinho et al., 2013, which stats that their provenience in this environment is still unclear. Antimicrobial resistance outcomes from the intensive use of antimicrobial drugs in human activities associated with various mechanisms for bacteria genetic transfer (Barbosa and Levy, 2000, Coque et al., 2008. Antibiotic-resistant bacteria enter into water environments where they are

块根块茎类作物内生固氮菌分离鉴定、系统发育与促生特性%Isolation, Identification, Phylogeny and Growth Promoting Characteristics of Endophytic Diazotrophs from Tuber and Root Crops

Institute of Scientific and Technical Information of China (English)

李艳星; 郭平毅; 孙建光

2017-01-01

. phosphorivorans, Flavimonas oryzihabitans, Flavobacterium oncorhynchi, Microbacterium hydrocarbonoxydans, Microbacterium phyllosphaerae, Micrococcus endophyticus, Paenibacillus barengoltzii, Pae. cineris, Pae. glycanilyticus, Pae. lautus, Pae. tundrae, Pantoea agglomerans, Pan. anthophila, Pan. dispersa, Pan. rodasii, Pseudomonas azotoformans, Ps. beteli, Ps. brassicacearum, Ps. geniculata, Ps. hunanensis, Ps. koreensis, Ps. parafulva, Ps. seleniipraecipitans, Ps. simiae, Ps. syringae, Rahnella aquatilis, Rhizobium leguminosarum, Rh. massiliae, Rh. radiobacter, Sphingobacterium canadense, Sp. faecium, Staphylococcus sciuri, Stenotrophomonas rhizophila, Variovorax paradoxus. This result showed the biodiversity of endophytic diazotrophs from tuber and root crops. Of the 219 endophytic diazotrophs, 77 strains are identified as 23 species of Bacillus, and 29 strains are identified as 10 species of Pseudomonas. This makes up 106 strains of 33 species, in percentages of 48.40% and 41.77% of the 219 endophytic diazotrophs and 79 identified species. Indicating that Bacillus and Pseudomonas are dominant populations of endophytic diazotrophs from tuber and root crops. Conducted with 79 representatives of the 219 strains, PGP test showed that 8.86% strains showed ACC deaminase activity ranging from 0.026 to 13.76 μmol α-ketobutyrate·mg-1 protein·h-1, 64.56% strains showed IAA production ranging from 0.34 to 28.99 μg·mL-1, and 6.33%-13.92% strains showed antagonistic against phytopathogen Fusarium sporotrichioides ACCC37402, Fusarium xysporum MLS1 and Fusarium xysporum ACCC37438 with antifungal indexes of 41% to 63%. [Conclusion] Large number of endophytic diazotrophs habitat in the tuber and root of normally growing tuber and root crops. Endophytic diazotrophs from tuber and root crops phylogenetically belong 79 species of 24 genera showing wide distribution and huge biodiversity. Bacillus and Pseudomonas are dominant populations of endophytic diazotrophs from tuber and root crops