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Sample records for aspergillus spp isolated

  1. Molecular identification of Aspergillus spp. isolated from coffee beans Identificação molecular de Aspergillus spp. isolados de grãos de café

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    Marciane Magnani

    2005-01-01

    Full Text Available Some species belonging to the genus Aspergillus are potential producers of ochratoxin A (OA, a mycotoxin with nephrotoxic, immunosuppressive, teratogenic and carcinogenic effects. The aim of the present study was to identify the species of Aspergillus that contaminate the inside of coffee beans collected in the stage of maturation and drying, from 16 producing areas located in the northern region of the State of Paraná, in the South of Brazil. A total of 108 isolates of Aspergillus spp. was identified at the species level, by sequencing the internal transcribed spacer (ITS1-5.8S-ITS2 of ribosomal DNA (rDNA. The results revealed the presence of potentially ochratoxigenic species in 82% of the geographic regions studied, among which Aspergillus niger was the species most frequently detected, followed by A. ochraceus and A. carbonarius. The presence of A. carbonarius in immature coffee fruits harvested from trees is reported for the first time.Algumas espécies pertencentes ao gênero Aspergillus possuem potencial para produção de Ocratoxina A (OA, uma micotoxina de efeitos nefrotóxicos, imunossupressivos, teratogênicos e carcinogênicos. Com o objetivo de identificar as espécies de Aspergillus que contaminam o interior de grãos de café, foram coletadas amostras em diferentes estádios de maturação do produto, em 16 propriedades produtoras do norte do estado do Paraná. Um total de 108 isolados de Aspergillus spp. foram identificados ao nível de espécie, pelo sequenciamento dos espaços internos transcritos (ITS1-5,8S-ITS2 do DNA ribossomal (rDNA. Os resultados revelaram a presença de espécies potencialmente ocratoxigênicas em 82% das regiões analisadas, sendo dentre estas, Aspergillus niger a espécie mais freqüentemente detectada,seguida por A. ochraceus, e A. carbonarius. É relatada pela primeira vez a presença de A. carbonarius em frutos de café coletados na árvore.

  2. In vitro synergy of natamycin and voriconazole against clinical isolates of Fusarium, Candida, Aspergillus and Curvularia spp.

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    Sradhanjali, Swatishree; Yein, Bandana; Sharma, Savitri; Das, Sujata

    2018-01-01

    To determine the minimum inhibitory concentrations (MICs) of voriconazole and natamycin, alone and in combination, against the clinical isolates of Fungus and to evaluate the synergy between the drugs in an experimental in vitro study. In an experimental in vitro study, clinical isolates of Fusarium , Aspergillus , Candida and Curvularia spp were maintained on Sabouraud Dextrose Agar and used for the study. The MICs of natamycin and voriconazole, used alone and in combination, were evaluated by checkerboard microdilution technique based on the standard protocol proposed by the Clinical Laboratory Standards Institute. The interactions were assessed using the fractional inhibitory concentration (FIC) Index model. Tested with all the clinical isolates, the MICs ranged between 0.125 and 8 µg/mL both for natamycin and voriconazole. In descending order, maximum synergism (FIC ≤0.5) was observed in Candida spp (33.3%) followed by Curvularia spp and Fusarium spp (23.1%). Synergism was least for Aspergillus spp (22.2%). However, at 61.5% (8/13), maximum additive effect (>0.5-1) was observed in Aspergillus spp and minimum (33.3%, 2/6) in Candida spp. Indifference (FIC value >1 and≤4) was observed in 22.2% (2/9) of Aspergillus spp, 15.4% (2/13) of Fusarium spp, 33.3% (2/6) of Candida spp and 23.1% (3/13) of Curvularia spp. No cases of antagonism (FIC >4) were observed. Natamycin and voriconazole in combination demonstrated more effective antifungal activity than single-use in vitro treatment in all species tested, which implies that these combinations may be helpful in treating fungal keratitis. There was no antagonism between these two drugs. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  3. Importance of Aspergillus spp. isolation in Acute exacerbations of severe COPD: prevalence, factors and follow-up: the FUNGI-COPD study

    Science.gov (United States)

    2014-01-01

    Background Acute exacerbations of COPD (AECOPD) are often associated with infectious agents, some of which may be non-usual, including Aspergillus spp. However, the importance of Aspergillus spp. in the clinical management of AECOPD still remains unclear. Objectives The aims of the study were to analyze the prevalence and risk factors associated with Aspergillus spp. isolation in AECOPD, and to investigate the associated clinical outcomes during a 1-year follow-up period. Methods Patients presenting with an AECOPD requiring hospitalization were prospectively included from four hospitals across Spain. Clinical, radiological and microbiological data were collected at admission and during the follow-up period (1, 6 and 12 months after discharge), and re-admissions and mortality data collected during the follow-up. Results A total of 240 patients with severe AECOPD were included. Valid sputum samples were obtained in 144 (58%) patients, and in this group, the prevalence of Aspergillus spp. isolation was 16.6% on admission and 14.1% at one-year follow-up. Multivariate logistic-regression showed that AECOPD in the previous year (OR 12.35; 95% CI, 1.9-29.1; p Aspergillus spp. isolation. Conclusions The main risk factors for Aspergillus spp. isolation were AECOPD in the previous year and concomitant isolation of Pseudomonas aeruginosa. However, although Aspergillus spp. is often isolated in sputum samples from patients with AECOPD, the pathogenic and clinical significance remains unclear. PMID:24517318

  4. Notable fibrolytic enzyme production by Aspergillus spp. isolates from the gastrointestinal tract of beef cattle fed in lignified pastures.

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    Flávia Oliveira Abrão

    Full Text Available Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA at 48 h was significantly higher other periods (P < 0.05. Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77 and xylanase (3.13 ±.091 than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively. Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets.

  5. Ohcratoxin producing Aspergillus spp. Isolated from tropical soils in Sarawak, Malaysia

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    J.S.S. Seelan

    2010-03-01

    Full Text Available Aspergillus strains isolated from tropical soils were selected for additional characterization and for ochratoxin analysis, which was determined by ELISA method and high performance liquid chromatography (HPLC profiles. Because of its great morphological variability and mycotoxin production availability, 18 isolates of Aspergillus species were selected for this study. Only two isolates of these tropical soils, A. sulphureus and A. carbonarius, showed positive results for ohcratoxin (OA in lower concentration (0.05-0.10 µg/ml. Ochratoxin production by these species was confirmed by high performance liquid chromatography. HPLC analysis for ochratoxin producing A. sulphureus and A. carbonarius showed retention time, Rt value = 4.417 and Rt value = 4.081 respectively.

  6. Genotypic and phenotypic diversity in tropical strains of Aspergillus spp. (section Circumdati) isolated from insects.

    Science.gov (United States)

    Moraes, Aurea; Holanda, Veronica; Zahner, Viviane

    2006-04-01

    The morphology, multilocus enzyme electrophoresis (MLEE), and RAPD-PCR profiles of a panel of 63 strains of Aspergilus section Circumdati, all isoloated from Brazilian insects, were examined. When compared to the descriptions reported in the literature, differences were observed in terms of colony diameter for the representatives studies. Numerical taxonomy based on data generated by MLEE identified two distinct subgroups among the A. ochraceus isolates. In addition, phosphoglucose isomerase (GPI-1) was detected only in A. sclerotiorum, while phosphofructokinase (FK-1) and acid phosphatase (ACP-2) were present only in strains of A. sulphureus, suggesting that these alleles (bands) could be used for species-specific detection. Using RAPD-PCR, species-specific molecular markers were identified for both A. petrakii and A. sulphureus. These results are important from the taxonomic viewpoint and may also be used in the design of screening programs for the isoloation of new strains.

  7. Genetic diversity of Aspergillus species isolated from onychomycosis and Aspergillus hongkongensis sp. nov., with implications to antifungal susceptibility testing.

    Science.gov (United States)

    Tsang, Chi-Ching; Hui, Teresa W S; Lee, Kim-Chung; Chen, Jonathan H K; Ngan, Antonio H Y; Tam, Emily W T; Chan, Jasper F W; Wu, Andrea L; Cheung, Mei; Tse, Brian P H; Wu, Alan K L; Lai, Christopher K C; Tsang, Dominic N C; Que, Tak-Lun; Lam, Ching-Wan; Yuen, Kwok-Yung; Lau, Susanna K P; Woo, Patrick C Y

    2016-02-01

    Thirteen Aspergillus isolates recovered from nails of 13 patients (fingernails, n=2; toenails, n=11) with onychomycosis were characterized. Twelve strains were identified by multilocus sequencing as Aspergillus spp. (Aspergillus sydowii [n=4], Aspergillus welwitschiae [n=3], Aspergillus terreus [n=2], Aspergillus flavus [n=1], Aspergillus tubingensis [n=1], and Aspergillus unguis [n=1]). Isolates of A. terreus, A. flavus, and A. unguis were also identifiable by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The 13th isolate (HKU49(T)) possessed unique morphological characteristics different from other Aspergillus spp. Molecular characterization also unambiguously showed that HKU49(T) was distinct from other Aspergillus spp. We propose the novel species Aspergillus hongkongensis to describe this previously unknown fungus. Antifungal susceptibility testing showed most Aspergillus isolates had low MICs against itraconazole and voriconazole, but all Aspergillus isolates had high MICs against fluconazole. A diverse spectrum of Aspergillus species is associated with onychomycosis. Itraconazole and voriconazole are probably better drug options for Aspergillus onychomycosis. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Antifungal Activity of Culture Filtrates and Organic Extracts of Aspergillus spp. against Pythium ultimum

    OpenAIRE

    Rania Aydi-Ben Abdallah; Marwa Hassine; Hayfa Jabnoun-Khiareddine; Rabiaa Haouala; Mejda Daami-Remadi

    2014-01-01

    Culture filtrates, chloroform and ethyl acetate extracts of nine isolates of Aspergillus spp. (A. niger, A. terreus, A. flavus and Aspergillus sp.), isolated from soil and compost, were tested for antifungal activity against Pythium ultimum the causal agent of the potato Pythium leak. Culture filtrates showed a significant antifungal activity at the different tested concentrations. Total inhibition of the pathogen was induced by the filtrate of CH8 of Aspergillus sp., used at 10% ...

  9. Detection of Salmonella spp., Candida albicans, Aspergillus spp., and Antimicrobial Residues in Raw and Processed Cow Milk from Selected Smallholder Farms of Zimbabwe

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    Tryness Anastazia Mhone

    2012-01-01

    Full Text Available A cross-sectional study was conducted to detect the presence of Salmonella spp., Candida albicans, Aspergillus spp., and antimicrobial residues in raw milk (n=120 and processed cow milk (n=20 from smallholder dairy farms from three sites in Zimbabwe. Culture and isolation of Salmonella spp., C. albicans, and Aspergillus spp. were performed using selective media, while antimicrobial residues were detected by a dye reduction test. No Salmonella, but C. albicans (17.5%; 21/120, Aspergillus spp. (0.8%; 1/120, and antimicrobial residues (2.5%; 3/120 were detected from raw milk. C. albicans was isolated from all three sites, while Aspergillus spp. and antimicrobial residues were detected from sites 1 and 3, respectively. From processed milk, only C. albicans (5% was isolated while Aspergillus spp. and antimicrobial residues were not detected. These results suggested low prevalence of Salmonella spp. and Aspergillus spp. and a relatively high prevalence of C. albicans in raw milk from the smallholder farms. The potential public health risks of C. albicans and the detected antimicrobial residues need to be considered. Thus, educating farmers on improving milking hygiene and storage of milk and establishing programmes for monitoring antimicrobial residues may help to improve the safety of milk from smallholder farms.

  10. Trichocomaceae: biodiversity of Aspergillus spp and Penicillium spp residing in libraries.

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    Leite, Diniz Pereira; Yamamoto, Ana Caroline Akeme; Amadio, Janaína Vasconcellos Ribeiro de Souza; Martins, Evelin Rodrigues; do Santos, Fábio Alexandre Leal; Simões, Sara de Almeida Alves; Hahn, Rosane Christine

    2012-10-19

    Atmospheric air is the most common vehicle for the dispersion of fungi. Fungi belonging to the genera Aspergillus and Penicillium are cosmopolitan and are classified in the family Trichocomaceae. Species of the genera are commonly found in soil, decaying organic materials, animal feed, stored grains, and other materials. This study aimed to determine the taxonomic diversity of airborne fungi of the genera Aspergillus and Penicillium residing in the dust of library environments to contribute to current knowledge of these characteristic genera. Three libraries in the city of Cuiaba, State of Mato Grosso, Brazil, were selected as the study areas. A total of 168 samples were collected at randomized sites within each library in areas containing journals, archives, in study rooms, and in collection storage areas in two different periods, the dry season (n = 42)  and the rainy season (n = 42). Samples were collected by exposing Petri dishes containing Sabouraud agar with chloramphenicol to the environmental air. Additional samples were collected with sterile swabs which were rubbed over the surface of randomly chosen books on the shelves; the swabs were subsequently incubated in the laboratory. The genus Aspergillus was highlighted as one of the principal airborne fungi present in indoor environments. Aspergillus spp was identified in 1,277 (89.6%) samples and Penicillium spp in 148 (10.4%). The dry period exhibited a greater number of isolates of the two taxons.

  11. Workflow to study genetic biodiversity of aflatoxigenic Aspergillus spp. in Georgia, USA

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    Peanut seeds were sampled from the entire state of Georgia in 2014. More than 600 isolates of Aspergillus spp. were collected using modified-dichloran rose Bengal (MDRB) medium, 240 of those isolates were fingerprinted with 25 InDel markers within the aflatoxin-biosynthesis gene cluster (ABC). Clust...

  12. Bacterial and Aspergillus spp. Contamination of Domestic Kitchens in Riyadh, Saudi Arabia

    International Nuclear Information System (INIS)

    Alwakeel, Suaad S.

    2007-01-01

    A randomized sampling of 50 households in Riyadh City, Saudi Arabia was conducted to determine microbial and Aspergillus spp contaminants in domestic kitchens between May and June 2006. Samples were taken from open air in the kitchen and from used kitchen sponges. Inoculation procedures were varied from direct inoculation of the sponge into the medium to dilution of a cut portion of the sponge. A total of 200 samples were taken from which, 700 culture plates were done (BAP and Nutrient agar). Identification by the API system of identification (Analytical Profile Index, BioMerieux) revealed Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus epidermidis, Enterobacter cloacae, Diphtheroids and Bacillus cereus, Aspergillus spp. was isolated and identified microscopically. Among the isolates, Staphylococcus epidermidis , Staphylococcus aureus was isolated in 90% of the plates followed by Pseudomonas aeruginosa (83%) , Klebsiella pneumonia ; Bacillus cereus (63%).and Aspergillus spp (15%) These opportunistic pathogens may be harmful especially in immunocompromised hosts. In this setting, there is a constant risk of contamination and transfer to willing hosts, thus appropriate measures should be implemented such as the use of disposable sponges. (author)

  13. Aspergillus saccharolyticus sp. nov., a new black Aspergillus species isolated in Denmark

    DEFF Research Database (Denmark)

    Sørensen, Annette; Lübeck, Peter S.; Lübeck, Mette

    2011-01-01

    A novel species, Aspergillus saccharolyticus sp. nov., belonging to the Aspergillus section Nigri group is described. This species was isolated in Denmark from treated hardwood. Its taxonomic status was determined using a polyphasic taxonomic approach including phenotypic (morphology and extrolite...... Aspergillus species that is morphologically similar to Aspergillus japonicus and Aspergillus aculeatus, but has a totally different extrolite profile compared to any known Aspergillus species. The type strain of A. saccharolyticus sp. nov. is CBS 127449T ( = IBT 28509T)....

  14. Production of aflatoxins by Aspergillus flavus and of fumonisins by Fusarium species isolated from Brazilian sorghum Avaliação da toxigenidade das cepas de Aspergillus flavus e Fusarium spp. isoladas de amostras de sorgo

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    Josefa B. da Silva

    2004-09-01

    Full Text Available Fifty-nine Aspergillus flavus and 35 Fusarium verticillioides strains, isolated from freshly harvested (10 and stored (130 Brazilian sorghum samples, were tested regarding their ability to produce aflatoxins (coconut milk agar and fumonisins (rice culture, respectively. Aflatoxins B1 and B2 were detected by TLC, and fumonisins B1 and B2 were analyzed by HPLC. Thirty-eight (64.4% A. flavus strains produced detectable levels of aflatoxins at concentrations ranging from 12.00 to 3282.50 µg/kg (AFB1 + AFB2, while thirty two (91% F. verticillioides strains produced FB1 at concentrations ranging from 0.12 to 5.38 µg/g. Two F. proliferatum strains produced low fumonisin levels. The toxigenic potential of A. flavus (64.4% and F. verticillioides (91.5% strains observed in sorghum samples indicates that rigorous control should be directed at the storage conditions of these products to minimize contamination with toxigenic deteriorating fungi, preventing further hazard to human and animal health.A produção de aflatoxinas por 59 cepas de Aspergillus flavus e fumonisinas por 35 cepas de Fusarium verticillioides isoladas de amostras de grãos de sorgo recém colhido (10 amostras e armazenado (130 amostras, foram avaliadas. A detecção de aflatoxinas (AFB1 e AFB2 foi efetuada por Cromatografia em Camada Delgada (CCD e fumonisinas (FB1 e FB2 foram analisadas por Cromatografia Líquida de Alta Eficiência (CLAE. Os resultados demonstram a produção de AFB1 e AFB2 em 38 cepas (64,4% de A. flavus cujos níveis variaram de 12,00 a 3282,50 µg/kg. Referente às cepas de F. verticillioides, 32 (91% produziram FB1, nas concentrações de 0,12 a 5,38 µg/g. Baixos níveis de fumonisinas foram detectados em 2 cepas de F. proliferatum. A constatação da potencialidade toxígena das cepas de A. flavus (64,4% e de F. verticillioides (91,5% nesta investigação, revelam a importância da pesquisa de aflatoxinas e fumonisinas nas amostras de sorgo. Diante disto

  15. Molecular identification of Aspergillus and Eurotium species isolated from rice and their toxin-producing ability.

    Science.gov (United States)

    Yazdani, D; Zainal Abidin, M A; Tan, Y H; Kamaruzaman, S

    2011-01-01

    Thirty milled rice samples were collected from retailers in 4 provinces of Malaysia. These samples were evaluated for Aspergillus spp. infection by direct plating on malt extract salt agar (MESA). All Aspergillus holomorphs were isolated and identified using nucleotide sequences of ITS 1 and ITS 2 of rDNA. Five anamorphs (Aspergillus flavus, A. oryzae, A. tamarii, A. fumigatus and A. niger) and 5 teleomorphs (Eurotium rubrum, E. amstelodami, E. chevalieri, E. cristatum and E. tonophilum) were identified. The PCR-sequencing based technique for sequences of ITS 1 and ITS 2 is a fast technique for identification of Aspergillus and Eurotium species, although it doesn't work flawlessly for differentiation of Eurotium species. All Aspergillus and Eurotium isolates were screened for their ability to produce aflatoxin and ochratoxin A (OTA) by HPLC and TLC techniques. Only A. flavus isolate UPM 89 was able to produce aflatoxins B1 and B2.

  16. Development of novel Alicyclobacillus spp. isolation medium.

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    Chang, S; Kang, D-H

    2005-01-01

    To develop a new isolation medium with higher recovery rates of Alicyclobacillus spp. SK agar was developed with optimized incubation temperature, pH, acidulant, Tween 80 concentration and divalent cation addition. Results indicate that detection of Alicyclobacillus spp. by SK agar was significantly higher (P > 0.05) than those obtained by K agar, orange serum agar, and potato dextrose agar. Current media used for Alicyclobacillus spp. isolation still resulted in high numbers of false negative products. The sensitivity of SK agar to Alicyclobacillus spp. allows detection of low numbers of Alicyclobacillus spp. and also provides a more higher isolation results compared with currently used media. SK agar will be useful to the fruit juice industry to obtain more accurate numbers of contaminant Alicyclobacillus spp. With this media, false negative samples can be reduced, and the likelihood of exported products being rejected can be greatly reduced.

  17. Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards.

    Science.gov (United States)

    García-Cela, E; Crespo-Sempere, A; Ramos, A J; Sanchis, V; Marin, S

    2014-03-03

    The aim of this study was to evaluate the diversity of black aspergilli isolated from berries from different agroclimatic regions of Spain. Growth characterization (in terms of temperature and water activity requirements) of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger was carried out on synthetic grape medium. A. tubingensis and A. niger showed higher maximum temperatures for growth (>45 °C versus 40-42 °C), and lower minimum aw requirements (0.83 aw versus 0.87 aw) than A. carbonarius. No differences in growth boundaries due to their geographical origin were found within A. niger aggregate isolates. Conversely, A. carbonarius isolates from the hotter and drier region grew and produced OTA at lower aw than other isolates. However, little genetic diversity in A. carbonarius was observed for the microsatellites tested and the same sequence of β-tubulin gene was observed; therefore intraspecific variability did not correlate with the geographical origin of the isolates or with their ability to produce OTA. Climatic change prediction points to drier and hotter climatic scenarios where A. tubingensis and A. niger could be even more prevalent over A. carbonarius, since they are better adapted to extreme high temperature and drier conditions. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. A prospective survey of Aspergillus spp. in respiratory tract samples: prevalence, clinical impact and antifungal susceptibility

    DEFF Research Database (Denmark)

    Mortensen, K L; Johansen, H K; Fuursted, Kurt

    2011-01-01

    for routine microbiologic investigation were examined for Aspergillus following routine procedures and with extended incubation (5 days). Identification was done by morphologic criteria and susceptibility testing using EUCAST method for azoles and amphotericin B E-test. Invasive aspergillosis (IA......) was evaluated using modified EORTC/MSG criteria. A total of 11,368 airway samples were received. Growth of Aspergillus spp. was found in 129 and 151 patients using routine and extended incubation, respectively. Three patients had proven IA (2%), 11 probable (7%), four had allergic bronchopulmonary aspergillosis...... μg/ml (3/112 A. fumigatus, 1/2 A. terreus). In conclusion, Aspergillus appears to be an important pathogen in Denmark. Elevated itraconazole MICs were detected in 4% of the isolates including a multi-azole resistant isolate....

  19. Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Varga, János; Susca, Antonia

    2008-01-01

    uvarum sp. nov. isolates produced secalonic acid, common to other Aspergillus japonicus-related taxa, and geodin, erdin and dihydrogeodin, which are not produced by any other black aspergilli. None of the isolates were found to produce ochratoxin A. The novel species is most closely related to two......A novel species, Aspergillus uvarum sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on internal transcribed spacers (ITS), beta-tubulin and calmodulin gene sequences, by AFLP analysis and by extrolite profiles. Aspergillus...

  20. screening and improvement of local isolates of aspergillus niger

    African Journals Online (AJOL)

    DR. AMINU

    ABSTRACT. The study involved the screening of fourteen isolates of Aspergillus niger for citric acid production from glucose. The study was aimed at screening and improving local strains of Aspergillus niger with potential for citric acid production. All the isolates screened produced varying amounts of citric acid, the highest ...

  1. 21 CFR 866.3040 - Aspergillus spp. serological reagents.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3040 Aspergillus... consist of antigens and antisera used in various serological tests to identify antibodies to Aspergillus...

  2. MALDI-TOF MS Andromas strategy for the routine identification of bacteria, mycobacteria, yeasts, Aspergillus spp. and positive blood cultures.

    Science.gov (United States)

    Bille, E; Dauphin, B; Leto, J; Bougnoux, M-E; Beretti, J-L; Lotz, A; Suarez, S; Meyer, J; Join-Lambert, O; Descamps, P; Grall, N; Mory, F; Dubreuil, L; Berche, P; Nassif, X; Ferroni, A

    2012-11-01

    All organisms usually isolated in our laboratory are now routinely identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the Andromas software. The aim of this study was to describe the use of this strategy in a routine clinical microbiology laboratory. The microorganisms identified included bacteria, mycobacteria, yeasts and Aspergillus spp. isolated on solid media or extracted directly from blood cultures. MALDI-TOF MS was performed on 2665 bacteria isolated on solid media, corresponding to all bacteria isolated during this period except Escherichia coli grown on chromogenic media. All acquisitions were performed without extraction. After a single acquisition, 93.1% of bacteria grown on solid media were correctly identified. When the first acquisition was not contributory, a second acquisition was performed either the same day or the next day. After two acquisitions, the rate of bacteria identified increased to 99.2%. The failures reported on 21 strains were due to an unknown profile attributed to new species (9) or an insufficient quality of the spectrum (12). MALDI-TOF MS has been applied to 162 positive blood cultures. The identification rate was 91.4%. All mycobacteria isolated during this period (22) were correctly identified by MALDI-TOF MS without any extraction. For 96.3% and 92.2% of yeasts and Aspergillus spp., respectively, the identification was obtained with a single acquisition. After a second acquisition, the overall identification rate was 98.8% for yeasts (160/162) and 98.4% (63/64) for Aspergillus spp. In conclusion, the MALDI-TOF MS strategy used in this work allows a rapid and efficient identification of all microorganisms isolated routinely. © 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.

  3. Isolation and identification of fungi responsible for leaf spots disease ...

    African Journals Online (AJOL)

    The diseased plant leaves were taken to the laboratory for culture, isolation, and ... These included; Alternaria longipes, Aspergillus fumigatus, Aspergillus niger, ... Phomopsis mangiferae, Pseudofusicoccum spp. and Rhizopus oryzae.

  4. Aspergillus species isolated from mangrove forests in Borneo Island, Sarawak, Malaysia

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    J.S.S. Seelan

    2009-06-01

    Full Text Available A study on the occurrence of Aspergillus spp. on selected mangrove forests in Sarawak was conducted to find out their diversity and distribution. Samples were obtained from mangrove soils and leaf litters at different locations, i.e. Sematan, Lundu, Kampung Bako, Bako in Sarawak. Soil and leaf litter samples were taken randomly at different locations with five replicates from each area. A total of 138 isolates of Aspergillus species were obtained from the soil and leaf litter samples by using direct plating and Warcup method. Based on both macroscopic and microscopic observations, using an identification key, individual isolates were classified within the genus Aspergillus, belonging to three subgenera, four sections and five species. The fungi isolates were identified as A. terreus, A. flavipes, A. carneus, A. fumigatus and A. clavatus. The most frequent isolated species was A. flavipes (63.04%, followed by A. fumigatus (16.7%, A. terreus (13.04%, A. carneus (5.8% and A. clavatus (1.44%. All of the isolated Aspergillus species grew well on MEA and CYA at 25°C. A. carneus produced reddish sclerotia on MEA after seven days and this could be used as an important characteristic in this species identification. A. clavatus from mangrove soil in Kampung Bako has shown long conidiophores (ranging from 3-5 cm with swollen hyphal structures, while A. clavatus from Sematan area has shorter conidiophores (ranging from 2.5-3.5 cm on MEA.

  5. Biosynthesis of components with antifungal activity against Aspergillus spp. using Streptomyces hygroscopicus

    Directory of Open Access Journals (Sweden)

    Dodić Jelena M.

    2015-01-01

    Full Text Available Losses of apple fruit during storage are mainly caused by fungal phytopathogens. Traditionally, postharvest fungal disease is controlled by the application of synthetic fungicides. However, the harmful impact on environment as well as human health largely limits their application. To reduce these problems in agrochemicals usage, new compounds for plant protection, which are eco-friendly, should be developed. The aim of this study is optimization of medium composition in terms of glucose, soybean meal and phosphates content, by applying response surface methodology, for the production of agents with antifungal activity against Aspergillus spp. For biosynthesis was used strain of Streptomyces hygroscopicus isolated from the environment. Experiments were carried out in accordance with Box-Behnken design with three factors on three levels and three repetitions in the central point. Antifungal activity of the obtained cultivation mediums against Aspergillus oryzae and Aspergillus niger was determined, in vitro, using the diffusion - disc method. For determination optimal medium components desirability function was used. Achieved model predicts that the maximum inhibition zone diameter (40.93 mm against test microorganisms is produced when the initial content of glucose, soybean meal and phosphates are 47.77 g/l, 24.54 g/l and 0.98 g/l, respectively. To minimize the consumption of medium components and costs of effluents processing, additional three sets of optimization were made. The chosen method for optimization of medium components was efficient, relatively simple and time and material saving. Obtained results can be used for the further techno-economic analysis of the process to select optimal medium composition for industrial application.

  6. Production of aspartic peptidases by Aspergillus spp. using tuna ...

    African Journals Online (AJOL)

    The production of extracellular aspartic peptidase by the fungi Aspergillus niger and Aspergillus awamori was carried out in a shake flask and in stirred tank submerged fermentations using tuna cooked wastewater, an industrial effluent, as nitrogen source for culture medium. In stirred tank fermentation, biomass production ...

  7. Aspergillus saccharolyticus sp. nov., a new black Aspergillus species isolated in Denmark

    DEFF Research Database (Denmark)

    Sørensen, Annette; Lübeck, Peter S.; Lübeck, Mette

    2011-01-01

    A novel species, Aspergillus saccharolyticus sp. nov., belonging to the Aspergillus section Nigri group is described. This species was isolated in Denmark from treated hardwood. Its taxonomic status was determined using a polyphasic taxonomic approach including phenotypic (morphology and extrolite...... profiles) and molecular (β-tubulin, internal transcribed spacer and calmodulin gene sequences, and universally primed PCR fingerprinting) analysis. Phenotypic and molecular data enabled this novel species to be clearly distinguished from other black aspergilli. A. saccharolyticus is a uniseriate...

  8. Aspergillus pragensis sp nov discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi

    DEFF Research Database (Denmark)

    Lyskova, Pavlina; Hubka, Vit; Kolarik, Miroslav

    2014-01-01

    The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of beta-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspe...

  9. Characterization of toxigenic and atoxigenic Aspergillus flavus isolates from pistachio

    Science.gov (United States)

    Thirty eight Aspergillus flavus isolates collected from a pistachio orchard in California were analyzed for production of aflatoxin (AF), cyclopiazonic acid (CPA), vegetative compatibility groups (VCGs) and mating types. All toxigenic isolates produced both AFB1 and CPA. Twenty-one percent of the i...

  10. Detection of Polish clinical Aspergillus fumigatus isolates resistant to triazoles

    DEFF Research Database (Denmark)

    Nawrot, Urszula; Kurzyk, Ewelina; Arendrup, Maiken Cavling

    2018-01-01

    We studied the presence of triazole resistance of 121 Aspergillus fumigatus clinical isolates collected in two Polish cities, Warsaw and Wrocław, to determine if resistance is emerging in our country. We identified five itraconazole resistant isolates (4.13%) carrying the TR34/L98H alteration...

  11. Detection of Aspergillus spp . and determination of the levels of ...

    African Journals Online (AJOL)

    Aflatoxins (B1, B2, G1 and G2) are hepatotoxic metabolites produced by Aspergillus flavus on a number of agricultural commodities. Their levels were studied in rice samples imported to Iran through a southern port in Bushehr. Aflatoxins analysis was performed by solvent extraction, immunoaffinity clean-up and ...

  12. Triazole Resistance in Aspergillus spp.: A Worldwide Problem?

    Science.gov (United States)

    Rivero-Menendez, Olga; Alastruey-Izquierdo, Ana; Mellado, Emilia; Cuenca-Estrella, Manuel

    2016-01-01

    Since the first description of an azole-resistant A. fumigatus strain in 1997, there has been an increasing number of papers describing the emergence of azole resistance. Firstly reported in the USA and soon after in Europe, it has now been described worldwide, challenging the management of human aspergillosis. The main mechanism of resistance is the modification of the azole target enzyme: 14-α sterol demethylase, encoded by the cyp51A gene; although recently, other resistance mechanisms have also been implicated. In addition, a shift in the epidemiology has been noted with other Aspergillus species (mostly azole resistant) increasingly being reported as causative agents of human disease. This paper reviews the current situation of Aspergillus azole resistance and its implications in the clinical setting. PMID:29376938

  13. In vitro susceptibility testing of Aspergillus spp. against voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin.

    Science.gov (United States)

    Shi, Jun-yan; Xu, Ying-chun; Shi, Yi; Lü, Huo-xiang; Liu, Yong; Zhao, Wang-sheng; Chen, Dong-mei; Xi, Li-yan; Zhou, Xin; Wang, He; Guo, Li-na

    2010-10-01

    During recent years, the incidence of serious infections caused by opportunistic fungi has increased dramatically due to alterations of the immune status of patients with hematological diseases, malignant tumors, transplantations and so forth. Unfortunately, the wide use of triazole antifungal agents to treat these infections has lead to the emergence of Aspergillus spp. resistant to triazoles. The present study was to assess the in vitro activities of five antifungal agents (voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin) against different kinds of Aspergillus spp. that are commonly encountered in the clinical setting. The agar-based Etest MIC method was employed. One hundred and seven strains of Aspergillus spp. (5 species) were collected and prepared according to Etest Technique Manuel. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48 hours at 35°C. MIC(50), MIC(90) and MIC range were acquired by Whonet 5.4 software. The MIC(90) of caspofungin against A. fumigatus, A. flavus and A. nidulans was 0.094 µg/ml whereas the MIC(90) against A. niger was 0.19 µg/ml. For these four species, the MIC(90) of caspofungin was the lowest among the five antifungal agents. For A. terrus, the MIC(90) of posaconazole was the lowest. For A. fumigatus and A. flavus, the MIC(90) in order of increasing was caspofungin, posaconazole, voriconazole, itraconazole, and amphotericin B. The MIC of amphotericin B against A. terrus was higher than 32 µg/ml in all 7 strains tested. The in vitro antifungal susceptibility test shows the new drug caspofungin, which is a kind of echinocandins, has good activity against the five species of Aspergillus spp. and all the triazoles tested have better in vitro activity than traditional amphotericin B.

  14. Citric acid production from whey by fermentation using Aspergillus spp.

    Directory of Open Access Journals (Sweden)

    Óscar Julián Sánchez Toro

    2004-01-01

    Full Text Available Whey has become the main dairy-industry waste product, despite continuous efforts aimed at finding a way to use it. The aim of this research was to investigate citric acid production by submerged fermentation using Aspergillus genus fungi, using whey as substrate to take economical advantage of it and to reduce the environmental impact caused by discharging this by-product into nearby streams. The following three strains were used: A. carbonarius NRRL 368, A. carbonarius NRRL 67 and A. niger NRRL 3. The best adaptation medium for inoculum propagation was selected. Proposed experimental design for evaluating citric acid biosynthesis from whey modified through different treatments showed that the two A. carbonarius strains did not present significant differences in acid production whereas A. niger NRRL 3 reached higher concentration when evaporated, deproteinised and p-galactosidase lactose-hydrolysed whey was used. However, A. carbonarius gave higher average citric acid titres than those found for A. niger. This suggests the need for carrying out further research on it as a potential producing strain. Cell growth, substrate consumption and acid production kinetics in a 3-L stirred-tank bioreactor with aeration were developed in the case of A. niger; kinetics were simulated through non-structured mathematical models. Key words: Aspergilluscarbonarius, Aspergillus niger, bioreactor, simulation, p-galactosidase.

  15. Phenotypic characterization of canine Malassezia spp., isolates

    Directory of Open Access Journals (Sweden)

    Angélica Hurtado-Suárez

    2016-09-01

    Full Text Available Objective. To characterize and identify yeasts of the genus Malassezia by phenotypic features. Materials and methods. First, the macroscopic and microscopic morphological characteristics were described. In addition we performed biochemical and physiological assays as Tweens and Cremophor, including more. Results. Our results evidenced of 105 isolates obtained from dogs diagnosed with external otitis, it was possible to identify two distinct species from 46 isolates within the Malassezia genus: 36.19% (n=38 were identified as M. pachydermatis and 7.62% (n=8 as M. furfur. According to phenotypic patterns the remaining 56.19% (n=59 were reported as Malassezia spp., possibly corresponding to M. furfur and/or M. pachydermatis. Conclusions. Results emphasize the necessity to characterize according to species. It is not feasible to define Malassezia by species based on morphological, biochemical, and physiological findings. Therefore, molecular genotyping should be performed to identify markers allowing a more precise isolate identification. This would broaden our epidemiological knowledge regarding different species involved in canine otitis pathologies.

  16. In vitro activity of the novel antifungal compound F901318 against difficult-to-treat Aspergillus isolates.

    Science.gov (United States)

    Buil, J B; Rijs, A J M M; Meis, J F; Birch, M; Law, D; Melchers, W J G; Verweij, P E

    2017-09-01

    F901318 is a new antifungal agent with a novel mechanism of action with activity against Aspergillus species. We investigated the in vitro activity of F901318 against a collection of Aspergillus isolates. A total of 213 Aspergillus isolates were used in this study. A total of 143 Aspergillus fumigatus sensu stricto isolates were used, of which 133 were azole resistant [25 TR34/L98H; 25 TR46/Y121F/T289A; 33 A. fumigatus with cyp51A-associated point mutations (25 G54, 1 G432 and 7 M220); and 50 azole-resistant A. fumigatus without known resistance mechanisms]. Ten azole-susceptible A. fumigatus isolates were used as WT controls. The in vitro activity was also determined against Aspergillus calidoustus (25 isolates), Aspergillus flavus (10), Aspergillus nidulans (10) and Aspergillus tubingensis (25). F901318 activity was compared with that of itraconazole, voriconazole, posaconazole, isavuconazole, amphotericin B and anidulafungin. Minimum effective concentrations and MICs were determined using the EUCAST broth microdilution method. F901318 was active against all tested isolates: A. fumigatus WT, MIC90 0.125 mg/L (range 0.031-0.125); TR34/L98H,TR46/Y121F/T289A and azole resistant without known resistance mechanisms, MIC90 0.125 mg/L (range 0.031-0.25); A. fumigatus with cyp51A-associated point mutations, MIC90 0.062 mg/L (range 0.015-0.125); and other species, A. calidoustus MIC90 0.5 mg/L (range 0.125-0.5), A. flavus MIC90 0.062 mg/L (range 0.015-0.62), A. nidulans MIC90 0.125 mg/L (range 0.062-0.25) and A. tubingensis MIC90 0.062 mg/L (range 0.015-0.25). F901318 showed potent and consistent in vitro activity against difficult-to-treat Aspergillus spp. with intrinsic and acquired antifungal resistance due to known and unknown resistance mechanisms, suggesting no significant implications of azole resistance mechanisms for the mode of action of F901318. © The Author 2017. Published by Oxford University Press on behalf of the British Society for

  17. [Respiratory infections caused by Aspergillus spp. in critically ill patients admitted to the intensive care units].

    Science.gov (United States)

    Álvarez Lerma, F; Olaechea Astigarraga, P; Palomar Martínez, M; Rodríguez Carvajal, M; Machado Casas, J F; Jiménez Quintana, M M; Esteve Urbano, F; Ballesteros Herráez, J C; Zavala Zegarra, E

    2015-04-01

    The presence of respiratory fungal infection in the critically ill patient is associated with high morbidity and mortality. To assess the incidence of respiratory infection caused by Aspergillus spp. independently of the origin of infection in patients admitted to Spanish ICUs, as well as to describe the rates, characteristics, outcomes and prognostic factors in patients with this type of infection. An observational, retrospective, open-label and multicenter study was carried out in a cohort of patients with respiratory infection caused by Aspergillus spp. admitted to Spanish ICUs between 2006 and 2012 (months of April, May and June), and included in the ENVIN-HELICS registry (108,244 patients and 825,797 days of ICU stay). Variables independently related to in-hospital mortality were identified by multiple logistic regression analysis. A total of 267 patients from 79 of the 198 participating ICUs were included (2.46 cases per 1000 ICU patients and 3.23 episodes per 10,000 days of ICU stay). From a clinical point of view, infections were classified as ventilator-associated pneumonia in 93 cases (34.8%), pneumonia unrelated to mechanical ventilation in 120 cases (44.9%), and tracheobronchitis in 54 cases (20.2%). The study population included older patients (mean 64.8±17.1 years), with a high severity level (APACHE II score 22.03±7.7), clinical diseases (64.8%) and prolonged hospital stay before the identification of Aspergillus spp. (median 11 days), transferred to the ICU mainly from hospital wards (58.1%) and with high ICU (57.3%) and hospital (59.6%) mortality rates, exhibiting important differences depending on the type of infection involved. Independent mortality risk factors were previous admission to a hospital ward (OR=7.08, 95%CI: 3.18-15.76), a history of immunosuppression (OR=2.52, 95%CI: 1.24-5.13) and severe sepsis or septic shock (OR=8.91, 95%CI: 4.24-18.76). Respiratory infections caused by Aspergillus spp. in critically ill patients admitted to

  18. Influence of gamma irradiation on natural mycoflora of poultry feed and effect on morphology, physiology and genetic of strains Aspergillus spp

    International Nuclear Information System (INIS)

    Ribeiro, Jessika Mara Martins

    2008-01-01

    Maize flour samples, soy crumb and feed were collected directly from the production line of a poultry farm in Avelar, RJ, and exposed to doses of 0,3.5, 0,8 and 15 kGy of gamma irradiation. Counting, isolation and identification of the contaminant mycoflora were performed before and after irradiation. The radiosensitivity of strains of reference of Aspergillus spp. was determined in CYA medium and in corn for doses ranging from 0 to 8 kGy. Comparison between the morphologies of control and irradiated strains were performed by using macroscopy, optical microscopy and transmission electron microscopy. Toxigenic profile determination and genetic evaluation by RAPD were also carried out. Higher doses have been found to reduce the number of active colonies, causing elimination of the mycoflora at 8 kGy. A larger radiosensitivity of yeasts was observed in comparison with filamentous fungi. A significant reduction in fungi population occurred at 3.5 kGy to levels below the limit that ensures the hygienic quality of ingredients and poultry feeds. The residual mycoflora was found to decrease with post-irradiation time and included mostly Cladosporium spp., Curvularia spp., Fusarium spp. and Aspergillus spp. and sterility of mycelium prevented further identification of the surviving species of Aspergillus spp. Differences in radioresistance were found among species of Aspergillus and the highest tolerance to radiation was observed for A. parasiticus. Initial morphologic changes were found to be more severe during the first isolation after irradiation than in later ones, with the fungi gradually recovering their normal growth rate. Ultrastructural changes in the irradiated strains were observed mostly in the plasmatic membrane and membranous organelles of nuclei and mitochondria. An increase in the rate of production of toxins by the irradiated strains has been found, however no significant alterations have been observed in their genotypes. Such findings apparently indicate

  19. Wheat straw lignin degradation induction to aromatics by por Aspergillus spp. and Penicillium chrysogenum

    Directory of Open Access Journals (Sweden)

    Baltierra-Trejo Eduardo

    2016-02-01

    Full Text Available Wheat straw is a recalcitrant agricultural waste; incineration of this material represents an important environmental impact. Different reports have been made regarding the use of the structural components of wheat straw, i.e. cellulose, hemicellulose and lignin; however, lignin has been less exploited because it is largely considered the recalcitrant part. Residual wheat straw lignin (REWSLI has a potential biotech-nological value if depolymerization is attained to produce aromatics. Ligninolytic mitosporic fungus represent an alternative where very little research has been done, even though they are capable of depol-ymerize REWSLI in simple nutritional conditions in relatively short periods, when compared to basidio-mycetes. The aim of this research was to study the depolymerization activity of Aspergillus spp and Penicillium spp on semipurified REWSLI as the sole carbon source to produce aromatics. The depoly-merization capacity was determined by the activity of the laccase, lignin peroxidase and manganese peroxidase enzymes. The generated aromatics derived from the REWSLI depolymerization were identi-fied by gas chromatography. Obtained results revealed that Penicillium chrysogenum depolymerized the lignin material by 34.8% during the 28-day experimentation period. Laccase activity showed the largest activity with 111 U L-1 in a seven-day period, this enzyme induction was detected in a smaller period than that required by basidiomycetes to induce it. Moreover, the enzymatic activity was produced with-out the addition of an extra carbon source as metabolic inductor. Aspergillus spp and Penicillium spp generated guaiacol, vanillin, and hydroxybenzoic, vanillinic, syringic and ferulic acid with a maximum weekly production of 3.5, 3.3, 3.2, 3.3, 10.1 and 21.9 mg mL-1, respectively.

  20. Retraction of colonies and structures of Aspergillus Spp. as a possible high dose sensor

    International Nuclear Information System (INIS)

    Borges, Valeria B.; Vital, Helio C.; Moraes, Aurea M.L.

    2013-01-01

    Changes in the length of macro-and microscopic structures of irradiated Aspergillus spp. colonies were investigated in this work in search for correlations with radiation dose. Effects related to growth and morphology produced by exposure to radiation doses ranging from 0.0 up to 8.0 kGy on strains of A. flavus (CMT 00079), A. parasiticus (CMT 00064) and A ochraceus (CMT 00145) were analyzed. Fungal colonies were inoculated on fresh peanut seeds and incubated for 5 days at 25°C. On the sixth day the samples were irradiated and moved to Petri dishes containing PDA culture medium where they remained incubated for 7 days at 25°C Post irradiation growth was periodically monitored by visual inspections and measurements of mycelial diameters. In addition, microscopic analyses were performed to determine the length of the major structures of the colonies. It has been found that irradiation causes a decrease in the diameter of colonies as well as in the length of microscopic structures of the fungus. The amount of retraction has been found to be strongly correlated to radiation dose. Such findings hint at the possibility of using in situ Aspergillus spp. colonies as high-dose living dosimeters in the event of massive radiation exposures. (author)

  1. Studies on Thiobacilli spp. isolated from sandy beaches of Kerala

    Digital Repository Service at National Institute of Oceanography (India)

    Gore, P.S.; Raveendran, O.; Unnithan, R.V.

    Occurrence, isolation and oxidative activity of Thiobacilli spp. from some sandy beaches of Kerala are reported. These organisms were encountered in polluted beaches and were dominant during monsoon in all the beaches...

  2. Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing.

    Science.gov (United States)

    Gitman, Melissa R; McTaggart, Lisa; Spinato, Joanna; Poopalarajah, Rahgavi; Lister, Erin; Husain, Shahid; Kus, Julianne V

    2017-07-01

    Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods. © Crown copyright 2017.

  3. Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing

    Science.gov (United States)

    Gitman, Melissa R.; McTaggart, Lisa; Spinato, Joanna; Poopalarajah, Rahgavi; Lister, Erin; Husain, Shahid

    2017-01-01

    ABSTRACT Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods. PMID:28404678

  4. Disinfection efficacy of chlorine and peracetic acid alone or in combination against Aspergillus spp. and Candida albicans in drinking water.

    Science.gov (United States)

    Sisti, Maurizio; Brandi, Giorgio; De Santi, Mauro; Rinaldi, Laura; Schiavano, Giuditta F

    2012-03-01

    The aim of the present study was to evaluate the fungicidal activity of chlorine and peracetic acid in drinking water against various pathogenic Aspergillus spp. and Candida albicans strains. A. nidulans exhibited the greatest resistance, requiring 10 ppm of chlorine for 30 min contact time for a complete inactivation. Under the same experimental conditions, peracetic acid was even less fungicidal. In this case, A. niger proved to be the most resistant species (50 ppm for 60 min for complete inactivation). All Aspergillus spp. were insensitive to 10 ppm even with extended exposure (>5 h). The combination of chlorine and peracetic acid against Aspergillus spp. did not show synergistic effects except in the case of A. flavus. Complete growth inhibition of C. albicans was observed after about 3 h contact time with 0.2 ppm. C. albicans was less sensitive to peracetic acid. Hence the concentrations of chlorine that are usually present in drinking water distribution systems are ineffective against several Aspergillus spp. and peracetic acid cannot be considered an alternative to chlorine for disinfecting drinking water. The combination of the two biocides is not very effective in eliminating filamentous fungi at the concentrations permitted for drinking water disinfection.

  5. Profiling of volatile organic compounds produced by clinical Aspergillus isolates using gas chromatography-mass spectrometry

    NARCIS (Netherlands)

    Gerritsen, M G; Brinkman, P; Escobar Salazar, Natalia; Bos, L D; de Heer, K; Meijer, M; Janssen, H-G; de Cock, H; Wösten, H A B; Visser, C.E.; van Oers, M H J; Sterk, P J

    Volatile organic compounds (VOCs) in exhaled breath may identify the presence of invasive pulmonary aspergillosis. We aimed to detect VOC profiles emitted by in vitro cultured, clinical Aspergillus isolates using gas chromatography-mass spectrometry (GC-MS). Three clinical Aspergillus isolates and a

  6. Profiling of volatile organic compounds produced by clinical Aspergillus isolates using gas chromatography-mass spectrometry

    NARCIS (Netherlands)

    Gerritsen, M. G.; Brinkman, P.; Escobar, N.; Bos, L. D.; de Heer, K.; Meijer, M.; Janssen, H.-G.; de Cock, H.; Wösten, H. A. B.; Visser, C. E.; van Oers, M. H. J.; Sterk, P. J.

    2018-01-01

    Volatile organic compounds (VOCs) in exhaled breath may identify the presence of invasive pulmonary aspergillosis. We aimed to detect VOC profiles emitted by in vitro cultured, clinical Aspergillus isolates using gas chromatography-mass spectrometry (GC-MS). Three clinical Aspergillus isolates and a

  7. Treatment of two postoperative endophthalmitis cases due to Aspergillus flavus and Scopulariopsis spp. with local and systemic antifungal therapy

    Directory of Open Access Journals (Sweden)

    Uyar Guliz

    2007-07-01

    Full Text Available Abstract Background Endophthalmitis is the inflammatory response to invasion of the eye with bacteria or fungi. The incidence of endophthalmitis after cataract surgery varies between 0.072–0.13 percent. Treatment of endophthalmitis with fungal etiology is difficult. Case Presentation Case 1: A 71-year old male diabetic patient developed postoperative endophthalmitis due to Aspergillus flavus. The patient was treated with topical amphotericin B ophthalmic solution, intravenous (IV liposomal amphotericin-B and caspofungin following vitrectomy. Case 2: A 72-year old male cachectic patient developed postoperative endophthalmitis due to Scopulariopsis spp. The patient was treated with topical and IV voriconazole and caspofungin. Conclusion Aspergillus spp. are responsible of postoperative fungal endophthalmitis. Endophthalmitis caused by Scopulariopsis spp. is a very rare condition. The two cases were successfully treated with local and systemic antifungal therapy.

  8. Antifungals susceptibility pattern of Candida spp . isolated from ...

    African Journals Online (AJOL)

    The highest susceptibility of the isolates was seen for nystatin 62 (83.78%), ketoconazole 61 (82.43%) and fluconazole 60 (81.08%). Conclusion: Despite the noticeable resistance of Candida spp. isolates to miconazole and itraconazole, the results indicate that nystatin, ketoconazole and fluconazole are the drugs of choice ...

  9. Molecular characterization of Azotobacter spp. nifH gene Isolated ...

    African Journals Online (AJOL)

    The Azotobacter species was isolated from marine source in two different seasons. They were cultivated under laboratory conditions using Nitrogen free Azotobacter specific medium. We observed that they were present in both seasons. The phylogenetic tree revealed that our isolated Azotobacter spp. was distantly related ...

  10. Influence of gamma irradiation on natural mycoflora of poultry feed and effect on morphology, physiology and genetic of strains Aspergillus spp;Influencia da radiacao gama sobre a micobiota natural de racao avicola e seu efeito sobre a morfologia, fisiologia e genetica de cepas de referencia de Aspergillus spp.

    Energy Technology Data Exchange (ETDEWEB)

    Ribeiro, Jessika Mara Martins

    2008-07-01

    Maize flour samples, soy crumb and feed were collected directly from the production line of a poultry farm in Avelar, RJ, and exposed to doses of 0,3.5, 0,8 and 15 kGy of gamma irradiation. Counting, isolation and identification of the contaminant mycoflora were performed before and after irradiation. The radiosensitivity of strains of reference of Aspergillus spp. was determined in CYA medium and in corn for doses ranging from 0 to 8 kGy. Comparison between the morphologies of control and irradiated strains were performed by using macroscopy, optical microscopy and transmission electron microscopy. Toxigenic profile determination and genetic evaluation by RAPD were also carried out. Higher doses have been found to reduce the number of active colonies, causing elimination of the mycoflora at 8 kGy. A larger radiosensitivity of yeasts was observed in comparison with filamentous fungi. A significant reduction in fungi population occurred at 3.5 kGy to levels below the limit that ensures the hygienic quality of ingredients and poultry feeds. The residual mycoflora was found to decrease with post-irradiation time and included mostly Cladosporium spp., Curvularia spp., Fusarium spp. and Aspergillus spp. and sterility of mycelium prevented further identification of the surviving species of Aspergillus spp. Differences in radioresistance were found among species of Aspergillus and the highest tolerance to radiation was observed for A. parasiticus. Initial morphologic changes were found to be more severe during the first isolation after irradiation than in later ones, with the fungi gradually recovering their normal growth rate. Ultrastructural changes in the irradiated strains were observed mostly in the plasmatic membrane and membranous organelles of nuclei and mitochondria. An increase in the rate of production of toxins by the irradiated strains has been found, however no significant alterations have been observed in their genotypes. Such findings apparently indicate

  11. Toxicity to Chicks of Aspergillus and Penicillium Species Isolated from Moldy Pecans 1

    Science.gov (United States)

    Doupnik, Ben; Bell, D. K.

    1971-01-01

    Isolates of Aspergillus chevalieri, A. flavus, A. ochraceus, A. repens, and Penicillium funiculosum and complexes of P. citrinum-P. implicatum isolated from moldy pecan meats were toxic to chicks. PMID:5564681

  12. Morpho-molecular identification of a novel aspergillus spp. and its cultural optimization for lipases production

    International Nuclear Information System (INIS)

    Iftikhar, T.; Niaz, M.; Haider, M.Z.; Sidra, A.

    2014-01-01

    Different lipid rich products were used to obtain oil degrading fungal isolates. The isolates were codified for referral to our culture bank and compared for their lipolytic potential. Amongst the isolates, MBL-1412 isolated from the cooked sliced cicer arietinum (Channa Daal) was found to be a potent hyper-producer and was optimized for lipase production under solid state fermentation. Initial systematic treatment based upon micrometric data and consultation with the standard monographs and fungus ended up with its identification as Aspergillus sp. The identification confirmed that the fungus belongs to genus Aspergillus, by DNA barcoding marker like 18S RNA gene sequence.Later, the sequence was registered with accession no. KM924434 in the public nucleotide library (genbank) of NCBI. Fungal culture was maintained on 2% potato dextrose agar (PDA) during the study. Diverse substrates of agricultural byproducts under varied incubation temperature, time interval, inoculum level and different pH of diluent were used as parameters of optimization for hyper-production of lipases. Different carbon and nitrogen sources as additives of culture medium were applied for enhancement of lipase production. Almond meal (10g) with inoculum level at 1.5 mL after 48 h of time course at 50 degree C and 6 pH were selected to be the best eco-cultural conditions for optimal lipases production by Aspergillus sp. MBL-1412. Supplementary additives of nitrogen and carbon sources to the basal substrate improved lipases production appreciably. Ammonium chloride (1%) as inorganic nitrogen source, nutrient broth (0.8%) as organic nitrogen source and starch (0.8%) as carbon source were found as best media additives for enhanced extracellular lipases yield. (author)

  13. Occurrence of Aspergillus spp. and aflatoxin B1 in Malaysian foods used for human consumption.

    Science.gov (United States)

    Reddy, Kasa R N; Farhana, Nazira I; Salleh, Baharuddin

    2011-05-01

    Malaysian population widely consumes the cereal-based foods, oilseeds, nuts, and spices in their daily diet. Mycotoxigenic fungi are well known to invade food products under storage conditions and produce mycotoxins that have threat to human and animal health. Therefore, determining toxigenic fungi and aflatoxin B(1) (AFB1) in foods used for human consumption is of prime importance to develop suitable management strategies and to minimize risk. Ninety-five food products marketed in Penang, Malaysia were randomly collected from different supermarkets and were analyzed for presence of Aspergillus spp. by agar plate assay and AFB1 by enzyme-linked immunosorbent assay (ELISA). A. flavus was the dominant fungi in all foods followed by A. niger. Fifty-five A. flavus strains were tested for their ability to produce aflatoxins on rice grain substrate. Thirty-six (65.4%) strains out of 55 produced AFB1 ranging from 1700 to 4400 μg/kg and 17 strains (31%) produced AFB2 ranging from 620 to 1670 μg/kg. Natural occurrence of AFB1 could be detected in 72.6% food products ranging from 0.54 to 15.33 μg/kg with a mean of 1.95 μg/kg. Maximum AFB1 levels were detected in peanut products ranging from 1.47 to 15.33 μg/kg. AFB1 levels detected in all food products were below the Malaysian permissible limits (<35 μg/kg). Aspergillus spp. and AFB1 was not detected in any cookies tested. Although this survey was not comprehensive, it provides valuable information on aflatoxin levels in foods marketed in Malaysia. © 2011 Institute of Food Technologists®

  14. Isolation of Cronobacter spp. (Enterobacter sakazakii from artisanal mozzarella

    Directory of Open Access Journals (Sweden)

    Francesco Casalinuovo

    2014-02-01

    Full Text Available Cronobacter spp. (Enterobacter sakazakii is an opportunistic bacterial pathogen capable of causing disease and even fatalities in newborn infants within the first weeks of life if consumed as part of the diet. Premature and immunocompromised newborn infants are at particular risk. The microorganism has been isolated from a variety of foods including contaminated infant milk formula powder and milk powder substitute. The study aimed to evaluate the level of microbiological contamination in 47 samples of mozzarella cheese made with cow’s milk collected from artisan cheese producers in Southern Italy. Samples were collected from commercial sales points and underwent qualitative and quantitative microbiological analyses to test for the bacterial contaminants most commonly found in milk and cheese products. The 47 samples underwent qualitative and quantitative microbiological tests according to ISO UNI EN standards. Analyses focused on Staphylococcus aures, Salmonella spp., Listeria monocytogenes, Pseudomonas spp., E. coli, Yersinia spp., total coliforms and Cronobacter sakazakii. The ISO/TS 22964:2006 method was used to investigate possible contamination by C. sakazakii. Biochemical identification was carried out using an automated system for identification and susceptibility tests. None of the samples examined resulted positive for Salmonella spp. or Listeria spp. Only one sample resulted positive for Staphylococcus aureus. Pseudomonas spp. was isolated in 10 (21% of 47 samples. High levels of total coliforms were found in 10 of 47 samples. Cronobacter spp. (Enterobacter sakazakii was isolated in one sample. This is the first study to confirm isolation of C. sakazakii in artisan mozzarella cheese made from cow’s milk. The presence of C. sakazakii could be related to external contamination during the phases of production or to the use of contaminated milk. Since mozzarella is recommended in the diet of children and adults of all ages, this

  15. A simple method for DNA isolation from Xanthomonas spp.

    Directory of Open Access Journals (Sweden)

    Gomes Luiz Humberto

    2000-01-01

    Full Text Available A simple DNA isolation method was developed with routine chemicals that yields high quality and integrity preparations when compared to some of the most well known protocols. The method described does not require the use of lysing enzymes, water bath and the DNA was obtained within 40 minutes The amount of nucleic acid extracted (measured in terms of absorbancy at 260 nm from strains of Xanthomonas spp., Pseudomonas spp. and Erwinia spp. was two to five times higher than that of the most commonly used method.

  16. Mycological survey of Korean cereals and production of mycotoxins by Fusarium isolates.

    OpenAIRE

    Lee, U S; Jang, H S; Tanaka, T; Toyasaki, N; Sugiura, Y; Oh, Y J; Cho, C M; Ueno, Y

    1986-01-01

    The fungal species isolated from Korean cereals (barley, polished barley, wheat, rye, and malt) were Alternaria spp., Aspergillus spp., Chaetomium spp., Drechslera spp., Epicoccum sp., Fusarium spp., and Penicillium spp., etc. The number of Fusarium strains isolated was 36, and their ability to produce Fusarium mycotoxins on rice was tested. Nivalenol (NIV) was produced by Fusarium graminearum (7 of 9 isolates), Fusarium oxysporum (3 of 10 isolates), and Fusarium spp. (7 of 15 isolates). Of 1...

  17. The genotypic characterization of Cronobacter spp. isolated in China.

    Directory of Open Access Journals (Sweden)

    Jinghua Cui

    Full Text Available Cronobacter spp. (Enterobacter sakazakii is an important pathogen contaminating powdered infant formula (PIF. To describe the genotypic diversity of Cronobacter isolated in China, we identified the isolates using fusA allele sequencing, and subtyped all of the isolates using pulsed-field gel electrophoresis (PFGE, multi-locus sequence typing (MLST, and multiple-locus variable-number tandem-repeat analysis (MLVA. A total of 105 isolates were identified, which included C. sakazakii (58 isolates, C. malonaticus (30 isolates, C. dublinensis (11 isolates, C. turicensis (5 isolates, and C. muytjensii (1 isolate. These isolates were showed to have 85 PFGE-patterns, 71 sequence types (STs, and 55 MLVA-patterns. Comparisons among the three molecular subtyping methods revealed that the PFGE method was the most distinguishable tool in identifying clusters of Cronobacter spp. through DNA fingerprinting, and MLST method came second. However, ESTR-1, ESTR-2, ESTR-3, and ESTR-4 were not effective loci for subtyping Cronobacter spp. such that the MLVA method requires further improvement.

  18. Biodegradation of HDPE by Aspergillus spp. from marine ecosystem of Gulf of Mannar, India.

    Science.gov (United States)

    Sangeetha Devi, Rajendran; Rajesh Kannan, Velu; Nivas, Duraisamy; Kannan, Kanthaiah; Chandru, Sekar; Robert Antony, Arokiaswamy

    2015-07-15

    High density polyethylene (HDPE) is the most commonly found non-degradable solid waste among the polyethylene. In this present study, HDPE degrading various fungal strains were isolated from the polyethylene waste dumped marine coastal area and screened under in vitro condition. Based on weight loss and FT-IR Spectrophotometric analysis, two fungal strains designated as VRKPT1 and VRKPT2 were found to be efficient in HDPE degradation. Through the sequence analysis of ITS region homology, the isolated fungi were identified as Aspergillus tubingensis VRKPT1 and Aspergillus flavus VRKPT2. The biofilm formation observed under epifluorescent microscope had shown the viability of fungal strains even after one month of incubation. The biodegradation of HDPE film nature was further investigated through SEM analysis. HDPE poses severe environmental threats and hence the ability of fungal isolates was proved to utilize virgin polyethylene as the carbon source without any pre-treatment and pro-oxidant additives. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Multicenter Study of Method-Dependent Epidemiological Cutoff Values for Detection of Resistance in Candida spp. and Aspergillus spp. to Amphotericin B and Echinocandins for the Etest Agar Diffusion Method

    DEFF Research Database (Denmark)

    Espinel-Ingroff, A; Arendrup, M; Cantón, E

    2017-01-01

    Method-dependent Etest epidemiological cutoff values (ECVs) are not available for susceptibility testing of either Candida or Aspergillus species with amphotericin B or echinocandins. In addition, reference caspofungin MICs for Candida spp. are unreliable. Candida and Aspergillus species wild-typ...

  20. Causative Agents of Aspergillosis Including Cryptic Aspergillus Species and A. fumigatus.

    Science.gov (United States)

    Toyotome, Takahito

    2016-01-01

    Aspergillosis is an important deep mycosis. The causative agents are Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, and Aspergillus terreus, of which A. fumigatus is the most prevalent. Cryptic Aspergillus spp., which morphologically resemble representative species of each Aspergillus section, also cause aspergillosis. Most of the cryptic species reveal different susceptibility patterns and/or different secondary metabolite profiles, also called exometabolome in this manuscript, from those representative species. On the other hand, azole-resistant A. fumigatus strains in clinical specimens and in the environment have been reported. Therefore, it is imperative to precisely identify the species, including cryptic Aspergillus spp., and evaluate the susceptibility of isolates.In this manuscript, some of the causative cryptic Aspergillus spp. are briefly reviewed. In addition, the exometabolome of Aspergillus section Fumigati is described. Finally, azole resistance of A. fumigatus is also discussed, in reference to several studies from Japan.

  1. Influence of Gamma-Irradiation on the Growth of Aspergillus spp. on Feeds for Ensuring Feed Safety

    International Nuclear Information System (INIS)

    Kim, J.S.; Woo, J.H.; Kim, J.T.; Park, N.H.; Kim, C.G.

    2010-01-01

    Aspergillus spp. is widely spread in the world on feeds and foods. They have been known to produce aflatoxins, which are mutagenic and carcinogenic to humans. The objective of this study was to determine the optimum gamma-irradiation dose for controling the growth of Aspergillus spp. to ensure safety of feeds. Four species fungal spore (10∨7 spore/mL) exposed to 0, 1, 3, 5, 7 and 10 kGy of gamma radiation were inhibited in their growth and AFBâ‚ productivity by greater-than or equal to 5 kGy. Meantime, the growth of fungal inoculated on feeds was inhibited at greater-than or equal to 3 kGy. However, AFBâ‚ presented in aqueous solution was not be inactivated completely by less-than or equal to 10 kGy irradiation. These results indicate that Aspergillus spp. on feeds could be controlled by 5 kGy gamma-irradiation but detoxification of AFBâ‚ demands a higher dose of gamma-irradiation (greater-than or equal to 10 kGy)

  2. Influence of Gamma-Irradiation on the Growth of Aspergillus spp. on Feeds for Ensuring Feed Safety

    International Nuclear Information System (INIS)

    Nam, B.R.; Kim, K.Y.; Ryu, H.J.

    2010-01-01

    Aspergillus spp. is widely spread in the world on feeds and foods. They have been known to produce aflatoxins, which are mutagenic and carcinogenic to humans. The objective of this study was to determine the optimum gamma-irradiation dose for controlling the growth of Aspergillus spp. to ensure safety of feeds. Four species fungal spore (10?7 spore/mL) exposed to 0, 1, 3, 5, 7 and 10 kGy of gamma radiation were inhibited in their growth and AFB? productivity by greater-than or equal to 5 kGy. Meantime, the growth of fungal inoculated on feeds was inhibited at greater-than or equal to 3 kGy. However, AFB? presented in aqueous solution was not be inactivated completely by less-than or equal to 10 kGy irradiation. These results indicate that Aspergillus spp. on feeds could be controlled by 5 kGy gamma-irradiation but detoxification of AFB? demands a higher dose of gamma-irradiation (greater-than or equal to 10 kGy)

  3. Antimicrobial activity of Streptomyces spp. Isolates from vegetable plantation soil

    Directory of Open Access Journals (Sweden)

    Isnaeni

    2016-05-01

    Full Text Available Fifteen Streptomyces isolates were isolated from soil in some different location on vegetable plantation at agriculture standard condition. The isolates were assessed for their antibacterial activity against Mycobacterium tuberculosis (MTB ATCC H37RV and mycobacterial which isolated from Dr. Soetomo Hospital patients in Surabaya. The International Streptomyces Project 4 (ISP4 and Middlebrook 7H9 (MB7H9 wwere used as growth or fermentation medium. The screening of inhibition activity was performed using turbidimetry and spot-test on agar medium. Results shown that 33.3% of the isolates (5 isolates have anti-mycobacterial activities. The first line anti tuberculosis drug rifampicin, (RIF, ethambutol (EMB, isoniazid (INH, and pyrazinamide (PZA were used as standards or positive controls with concentration 20 ppm. Optical density of crude fermentation broth concentrated from five isolates relatively lower than five anti-tuberculosis drug activity standard, although their activities against some microbial were similar to the standard at spot-test. The most efficient isolate shown anti-mycobacterial activity was Streptomyces B10 which identified as Streptomyces violaceousniger. In addition, fatty acid methyl ester (FAME profile of gas chromatography-mass spectrometry chromatogram of each isolates were studied and compared to Streptomyces spp. Keywords: Anti-mycobacterial, Mycobacterium tuberculosis, Streptomyces spp.

  4. Susceptibility testing of sequential isolates of Aspergillus fumigatus recovered from treated patients.

    NARCIS (Netherlands)

    Danaoui, E.; Meletiadis, J.; Tortorano, A.M.; Symoens, F.; Nolard, N.; Viviani, M.A.; Piens, M.A.; Lebeau, B.; Verweij, P.E.; Grillot, R.

    2004-01-01

    Two-hundred sequential Aspergillus fumigatus isolates recovered from 26 immunocompromised patients with invasive aspergillosis or bronchial colonization were tested for their in vitro susceptibility to posaconazole, itraconazole, voriconazole, terbinafine and amphotericin B. Twenty-one patients were

  5. Isolation, Optimization, and Investigation of Production of Linoleic Acid in Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Noushin Shafiei

    2016-08-01

    Full Text Available Background and Objectives: Microorganisms that are capable of accumulating lipid up to 20% of their biomass are called oleaginous microorganisms. In this study, optimization in lipid and linolenic acid production was investigated in Aspergillus niger as an oleaginous filamentous fungi. Methods: In this study, at first different strains of filamentous fungi were isolated, and after staining of the isolates with Sudan Black, their oil was extracted using chloroform/methanol. Then, the isolates with oil/dry biomass ratio of more than 20% were considered as oleaginous filamentous fungi. After microscopic examination, the identified isolate was optimized in terms of oil production. Finally, the amount of linolenic acid was evaluated using gas chromatography. Results: At first, 20 filamentous fungi isolates were isolated. According to the results of Sudan Black staining, lipid inclusions were observed in all the fungal isolates. The amount of oil produced in all isolates, showed that the percentage of oil production in isolates 4, 5, and 16, was more than 20%. In microscopic examination, the isolate 5 was Aspergillus niger. The best pH, temperature, time, and carbon source for oil production by Aspergillus niger was 4.5, 30°C, 96 hours, and fructose, respectively. The amount of linolenic acid in Aspergillus niger was reported 22.4% using gas chromatography.   Conclusion: The results of this study revealed that Aspergillus niger is an appropriate filamentous fungi for linolenic acid production.   

  6. Antimicrobial sensitivity profile of Staphylococcus spp. Isolated from clinical mastitis

    Directory of Open Access Journals (Sweden)

    Thamires Martins

    2012-12-01

    Full Text Available Inflammation of the mammary gland, which is also known as mastitis, occupies a prominent place among the diseases that affect dairy cattle, having a great economic importance in the dairy sector. Mastitis may have different origins, however, infectious mastitis is the most frequent and represents a risk to public health due to the propagation of microorganisms through milk. Staphylococcus spp. are considered the microorganisms that cause the greatest losses in milk production, being that Staphylococcus aureus is the pathogen of major importance because they present high resistence to antimicrobials. Empirical treatment, without prior identification of the pathogens and their resistance profile, may contribute to the emergence of multidrug-resistant strains and risk the efficiency of the antimicrobial. In that scenery, the study aimed to evaluate the resistance profile of Staphylococcus spp. against some antimicrobials used in the treatment of cows with clinical mastitis. The study was conducted on a property in the state of São Paulo from January 2011 to June 2012. We evaluated 29 lactating cows that present clinical mastitis in, at least, one mammary quarter. The diagnosis of clinical mastitis was performed by evaluating the clinical signs and also by Tamis test. Samples of milk from mammary quarters were collected aseptically in sterile tubes for microbiological evaluation. Microorganisms were isolated on sheep blood agar 5% and Sabouraud agar with chloramphenicol. The sensitivity profile of Staphylococcus spp. to the antibiotics ampicillin, cephalexin, ceftiofur, cefaclor, gentamicin, kanamycin, neomycin, penicillin G and oxacillin, was tested by disk diffusion test on Mueller-Hinton agar. From a total of 106 samples of milk analyzed, 64 (60.38% presented microbiological growth, being observed isolation of Streptococcus spp. 29 (34.52%, Staphylococcus spp. 28 (33.33%, Corynebacterium spp. 17 (20.24%, filamentous fungi 4 (4.76%, yeast 4 (4

  7. First Isolates of Leptospira spp., from Rodents Captured in Angola

    Science.gov (United States)

    Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa

    2016-01-01

    Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola. PMID:26928840

  8. First Isolates of Leptospira spp., from Rodents Captured in Angola.

    Science.gov (United States)

    Fortes-Gabriel, Elsa; Carreira, Teresa; Vieira, Maria Luísa

    2016-05-04

    Rodents play an important role in the transmission of pathogenic Leptospira spp. However, in Angola, neither the natural reservoirs of these spirochetes nor leptospirosis diagnosis has been considered. Regarding this gap, we captured rodents in Luanda and Huambo provinces to identify circulating Leptospira spp. Rodent kidney tissue was cultured and DNA amplified and sequenced. Culture isolates were evaluated for pathogenic status and typing with rabbit antisera; polymerase chain reaction (PCR) and sequencing were also performed. A total of 37 rodents were captured: Rattus rattus (15, 40.5%), Rattus norvegicus (9, 24.3%), and Mus musculus (13, 35.2%). Leptospiral DNA was amplified in eight (21.6%) kidney samples. From the cultures, we obtained four (10.8%) Leptospira isolates belonging to the Icterohaemorrhagiae and Ballum serogroups of Leptospira interrogans and Leptospira borgpetersenii genospecies, respectively. This study provides information about circulating leptospires spread by rats and mice in Angola. © The American Society of Tropical Medicine and Hygiene.

  9. Aspergillus pragensis sp nov discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi

    Czech Academy of Sciences Publication Activity Database

    Hubka, Vít; Lysková, P.; Frisvad, J.C.; Peterson, S.W.; Skořepová, M.; Kolařík, Miroslav

    2014-01-01

    Roč. 52, č. 6 (2014), s. 565-576 ISSN 1369-3786 R&D Projects: GA MŠk(CZ) EE2.3.20.0055; GA MŠk(CZ) EE2.3.30.0003 Institutional support: RVO:61388971 Keywords : Aspergillus candidus * Aspergillus tritici * antifungal susceptibility testing Subject RIV: EE - Microbiology, Virology Impact factor: 2.335, year: 2014

  10. CTX-M extended-spectrum β-lactamase-producing Klebsiella spp, Salmonella spp, Shigella spp and Escherichia coli isolates in Iranian hospitals.

    Science.gov (United States)

    Bialvaei, Abed Zahedi; Kafil, Hossein Samadi; Asgharzadeh, Mohammad; Aghazadeh, Mohammad; Yousefi, Mehdi

    2016-01-01

    This study was conducted in Iran in order to assess the distribution of CTX-M type ESBLs producing Enterobacteriaceae. From January 2012 to December 2013, totally 198 E. coli, 139 Klebsiella spp, 54 Salmonella spp and 52 Shigella spp from seven hospitals of six provinces in Iran were screened for resistance to extended-spectrum cephalosporins. After identification and susceptibility testing, isolates presenting multiple-drug resistance (MDR) were evaluated for ESBL production by the disk combination method and by Etest using (cefotaxime and cefotaxime plus clavulanic acid). All isolates were also screened for blaCTX-M using conventional PCR. A total of 42.92%, 33.81%, 14.81% and 7.69% of the E. coli, Klebsiella spp, Salmonella spp and Shigella spp isolates were MDR, respectively. The presence of CTX-M enzyme among ESBL-producing isolates was 85.18%, 77.7%, 50%, and 66.7%, in E. coli, Klebsiella spp, Salmonella spp and Shigella spp respectively. The overall presence of CTX-M genes in Enterobacteriaceae was 15.4% and among the resistant isolates was 47.6%. This study indicated that resistance to β-lactams mediated by CTX-M enzymes in Iran had similar pattern as in other parts of the world. In order to control the spread of resistance, comprehensive studies and programs are needed. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  11. ISOLATION AND PURIFICATION OF STREPTOMYCES SPP. PRODUCING VANCOMYCIN

    International Nuclear Information System (INIS)

    EL-KABBANY, H.M.I.

    2008-01-01

    Soil samples obtained from different governments in Egypt were analyzed to determine the presence of types of antibiotic producing actinomycetes using starch-nitrite agar, starch-casein nitrate agar and Czapek's Dox agar as culture media. Different Streptomyces spp. were isolated. The Streptomyces (S.) isolates encountered were S. violochromogens, S. violaceus-nigar and S. orientalis and known as standard Vancomycin producers. The optimum conditions of S. orientalis; incubation period, initial pH and incubation temperature, were determined. In addition, physical properties; appearance, melting point, solubility, mass spectrophotometer of ultra violet (UV) and the effect of gamma rays, were also determined

  12. Aflatoxin B1 producing potential of Aspergillus flavus strains isolated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... consumption of aflatoxin contaminated food and feed. (Reddy and ..... intervals by artificial inoculation of A. parasiticus on rice grains. ... additive for culture media for rapid identification of aflatoxin producing. Aspergillus strains ...

  13. Prevalence of potential toxigenic Aspergillus species isolated from ...

    African Journals Online (AJOL)

    ADEYEYE

    2015-12-17

    Dec 17, 2015 ... Aspergillus species) in feeds used in poultry farms in Sokoto metropolis. During a ... potential exists for the production of mycotoxins that may be of both veterinary public health significance and ..... The effect of antioxidant and.

  14. Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae)

    OpenAIRE

    Dali, Seniwati; Patong, A. B. D. Rauf; Jalaluddin, M. Noor; Pirman; Hamzah, Baharuddin

    2011-01-01

    Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum ...

  15. Xylanase production by a newly isolated Aspergillus niger SS7 in submerged culture.

    Science.gov (United States)

    Bakri, Yasser; Al-Jazairi, Manal; Al-Kayat, Ghassan

    2008-01-01

    Xylanase production by a newly isolated Aspergillus niger SS7 was studied in submerged culture. The optimum initial pH for xylanase production was found to be 7.0. Different agricultural and industrial wastes were evaluated for their ability to induce xylanase production by this isolate. The best xylanase production (293.82 IU/ml) was recorded at 3% (w/v) corn cob hulls after 120 h of incubation. The Aspergillus niger SS7 isolate grown in a simple medium, proved to be a promising microorganism for xylanase production.

  16. Atypical Aspergillus parasiticus isolates from pistachio with aflR gene nucleotide insertion identical to Aspergillus sojae

    Science.gov (United States)

    Aflatoxins are the most toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus flavus and Aspergillus parasiticus. The toxins cause devastating economic losses because of strict regulations on distribution of contaminated products. Aspergillus sojae are...

  17. Minimum inhibitory concentration distribution in environmental Legionella spp. isolates.

    Science.gov (United States)

    Sandalakis, Vassilios; Chochlakis, Dimosthenis; Goniotakis, Ioannis; Tselentis, Yannis; Psaroulaki, Anna

    2014-12-01

    In Greece standard tests are performed in the watering and cooling systems of hotels' units either as part of the surveillance scheme or following human infection. The purpose of this study was to establish the minimum inhibitory concentration (MIC) distributions of environmental Legionella isolates for six antimicrobials commonly used for the treatment of Legionella infections, by MIC-test methodology. Water samples were collected from 2004 to 2011 from 124 hotels from the four prefectures of Crete (Greece). Sixty-eight (68) Legionella isolates, comprising L. pneumophila serogroups 1, 2, 3, 5, 6, 8, 12, 13, 15, L. anisa, L. rubrilucens, L. maceachernii, L. quinlivanii, L. oakridgensis, and L. taurinensis, were included in the study. MIC-tests were performed on buffered charcoal yeast extract with α-ketoglutarate, L-cysteine, and ferric pyrophosphate. The MICs were read after 2 days of incubation at 36 ± 1 °C at 2.5% CO2. A large distribution in MICs was recorded for each species and each antibiotic tested. Rifampicin proved to be the most potent antibiotic regardless of the Legionella spp.; tetracycline appeared to have the least activity on our environmental isolates. The MIC-test approach is an easy, although not so cost-effective, way to determine MICs in Legionella spp. These data should be kept in mind especially since these Legionella species may cause human disease.

  18. Antifungal activity of indigenous Bacillus spp. isolated from soil

    Directory of Open Access Journals (Sweden)

    Bjelić Dragana Đ.

    2017-01-01

    Full Text Available Biocontrol using plant growth-promoting rhizobacteria (PGPR represents an alternative approach to disease management, since PGPR are known to promote growth and reduce diseases in various crops. Among the different PGPR, members of the genus Bacillus are prefered for most biotechnological uses due to their capability to form extremely resistant spores and produce a wide variety of metabolites with antimicrobial activity. The objective of this research was to identify antagonistic bacteria for management of the plant diseases. Eleven isolates of Bacillus spp. were obtained from the soil samples collected from different localities in the Province of Vojvodina. The antifungal activity of bacterial isolates against five fungal species was examined using a dual plate assay. Bacillus isolates exhibited the highest antifungal activity against Fusarium proliferatum, Fusarium oxysporum f. sp. cepae and Alternaria padwickii, while they had the least antagonistic effect on Fusarium verticillioides and Fusarium graminearum. Molecular identification showed that effective bacterial isolates were identified as Bacillus safensis (B2, Bacillus pumilus (B3, B11, Bacillus subtilis (B5, B7 and Bacillus megaterium (B8, B9. The highest antagonistic activity was exhibited by isolates B5 (from 39% to 62% reduction in fungal growth and B7 (from 40% to 71% reduction in fungal growth. These isolates of B. subtilis could be used as potential biocontrol agents of plant diseases. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073

  19. Cryptococcus spp isolated from dust microhabitat in Brazilian libraries

    Directory of Open Access Journals (Sweden)

    Leite Diniz P

    2012-06-01

    Full Text Available Abstract Background The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. Methods Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue, urea broth, and carbohydrate assimilation tests (auxanogram. Results Of the 84 samples collected from book dust, 18 (21.4% were positive for Cryptococcus spp totalizing 41 UFC’s. The most frequently isolated species was C. gattii 15 (36.6%; followed by C. terreus, 12 (29.3%; C. luteolus 4 (9.8%; C. neoformans, and C. uniguttulatus 3 (7.3%, and C. albidus and C. humiculus with 2 (4.6% of the isolates. Conclusion The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries.

  20. Cryptococcus spp isolated from dust microhabitat in Brazilian libraries.

    Science.gov (United States)

    Leite, Diniz P; Amadio, Janaina V R S; Martins, Evelin R; Simões, Sara A A; Yamamoto, Ana Caroline A; Leal-Santos, Fábio A; Takahara, Doracilde T; Hahn, Rosane C

    2012-06-08

    The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger) agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue), urea broth, and carbohydrate assimilation tests (auxanogram). Of the 84 samples collected from book dust, 18 (21.4%) were positive for Cryptococcus spp totalizing 41 UFC's. The most frequently isolated species was C. gattii 15 (36.6%); followed by C. terreus, 12 (29.3%); C. luteolus 4 (9.8%); C. neoformans, and C. uniguttulatus 3 (7.3%), and C. albidus and C. humiculus with 2 (4.6%) of the isolates. The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries.

  1. Glucoamylase production by a newly isolated strain of Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Sinkar, V.P.; Lewis, N.F.

    1982-01-01

    Glucoamylase production by Aspergillus niger 57 was studied in complex and synthetic media under stationary vs. submerged conditions. Stationary cultivation resulted in significantly greater yields than did submerged culture. Crude enzyme activity was optimum at 60 degrees and pH 4.0.

  2. Isolation of an Antioxidative Substance Produced by Aspergillus repens.

    Science.gov (United States)

    Yagi, R; Doi, M

    1999-01-01

    The acidic fraction of an extract of the culture liquid of Aspergillus repens MA0197 showed strong antioxidative activity when tested by the ferric thiocyanate and TBA methods. Chromatographic purification of this acidic fraction gave an active substance identified as Neoechinulin A. This compound showed higher antioxidative activity than α-tocopherol and could be expected to act as an antioxidant in Katsuobushi.

  3. Evaluation of the detection techniques of toxigenic Aspergillus isolates

    African Journals Online (AJOL)

    Aflatoxins are difuranocoumarin derivatives produced by some Aspergillus species through a polyketide pathway. Mycotoxins are considered hazardous and there is a need for accurate detection of each toxin. Several screening methods for direct visual determination of aflatoxins and ochratoxin A (OTA) production have ...

  4. Removal of trace element by isolates of Aspergillus brasiliensis ...

    African Journals Online (AJOL)

    Coffee beans processing generates a large volume of wastewater composed of trace elements which can be detrimental to human health. The present study aimed at evaluating the capacity of strains of Aspergillus brasiliensis and Penicillium citrinum in tolerating and removing trace elements namely: Cu, Mn and Zn from ...

  5. Cellulase Production by Aspergillus flavus Linn Isolate NSPR 101 ...

    African Journals Online (AJOL)

    Bagasse, corncob and sawdust were used as lignocellulosic substrates for the production of cellulase enzyme using Aspergillus flavus after ballmilling and pretreatment with caustic soda. From the fermentation studies, sawdust gave the best result with an enzyme activity value of 0.0743IU/ml while bagasse and corncob ...

  6. Isolation and antibiotic resistance of Ureaplasma spp. isolated from urogenital specimen between 2002 to 2007

    Directory of Open Access Journals (Sweden)

    Tito Del Gaudio

    2009-03-01

    Full Text Available Ureaplasma spp. and Mycoplasma hominis are frequently isolated from urogenital samples. Ureaplasma spp is responsible for cervicovaginitis, salpingitis, urethritis, epididymitis, male and female infertility, spontaneous abortion, and during pregnancy, for the premature rupture of the membranes, because of chorionamnionitis. Our study aimed to establish the pattern of antimicrobial resistance among Ureaplasma spp isolated in the area of Andria,Apulia Region, from January 2002 to December 2007. 240/781 (30.7% of the urogenital samples examined were found Ureaplasma spp.-positive. 152/240 (63.3 % were >104 UFC/ml and 88/240 (36.7 % were <104 UFC/ml. With regard to the resistance rate, we observed significant increase in resistance to ciprofloxacin, ofloxacin, erythromycin, clarithromycin, and azithromycin. While we did not observe resistance to doxycycline, strains resistant to tetracycline, josamycin, and pristinamycins, were isolated during last years of investigation. Our data may help improve the management of these infections above all in consideration of the differences among isolates in different geographic regions.

  7. Abundance and antibiotic susceptibility of Vibrio spp. isolated from microplastics

    Science.gov (United States)

    Laverty, A. L.; Darr, K.; Dobbs, F. C.

    2016-02-01

    In recent years, there has been a growing concern for `microplastics' (particles pieces, paired seawater samples, and from them cultured 44 putative Vibrio spp. isolates, 18 of which were PCR-confirmed as V. parahaemolyticus and 3 as V. vulnificus. There were no PCR-confirmed V. cholerae isolates. We used the Kirby-Bauer disk diffusion susceptibility test to examine the isolates' response to six antibiotics: chloramphenicol (30μg), gentamicin (10μg), ampicillin (10μg), streptomycin (10μg), tetracycline (30μg), and rifampin (5μg). Vibrio isolates were susceptible to three or more of the six antibiotics tested and all were susceptible to tetracycline and chloramphenicol. There were no apparent differences between the antibiotic susceptibilities of vibrios isolated from microplastics compared to those from the water column. In every instance tested, vibrios on microplastics were enriched by at least two orders of magnitude compared to those from paired seawater samples. This study demonstrates that microplastic particles serve as a habitat for Vibrio species, in particular V. vulnificus and V. parahaemolyticus, confirming the conjecture of Zettler et al. (2013) that plastics may serve as a vector for these and other potentially pathogenic bacteria.

  8. Assessment of Genetic Diversity among Pleurotus spp. Isolates from Jordan

    Directory of Open Access Journals (Sweden)

    Hanan Aref Hasan

    2018-04-01

    Full Text Available Pleurotus is considered an important genus that belongs to the family Pleurotaceae and includes the edible King Oyster mushroom (Pleurotus eryngii. In the present study, 19 Pleurotus isolates were collected from two locations in the north of Jordan (Tell ar-Rumman and Um-Qais. The morphological characteristics among collected isolates revealed that there was a morphological similarity among the collected isolates. Nucleotide sequence analysis of the internal transcribed spacer (ITS1–5.8S rDNA–ITS4 region and 28S nuclear large subunit (nLSU in the ribosomal DNA gene of the isolated stains showed that all of them share over 98% sequence similarity with P. eryngii. Genetic diversity among the collected strains was assessed using inter simple sequence repeat (ISSR analysis using 18 different primer pairs. Using this approach, 141 out of 196 bands obtained were considered polymorphic and the highest percentage of polymorphism was observed using primer UBC827 (92.3% with an overall Polymorphism Information Content (PIC value of 70.56%. Cluster analysis showed that the Jordanian Pleurotus isolates fall into two main clades with a coefficient of similarity values ranging from 0.59 to 0.74 with a clear clustering based on collection sites. The results of the present study reveal that molecular techniques of ISSR and rDNA sequencing can greatly aid in classification and identification of Pleurotus spp. in Jordan.

  9. Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae

    Directory of Open Access Journals (Sweden)

    Seniwati Dali

    2011-01-01

    Full Text Available Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum temperature, thermal stability and reusability were carried out. The results showed that free lipase had optimum pH 8,2 and optimum temperature 35 °C while the immobilized lipase had optimum 8,2 and optimum temperature 45 °C. The thermal stability of the immobilized lipase, relative to that of the free lipase, was markedly increased. The immobilized lipase can be reused for at least six times.

  10. The effect of high-power ultrasound and gas phase plasma treatment on Aspergillus spp. and Penicillium spp. count in pure culture.

    Science.gov (United States)

    Herceg, Z; Režek Jambrak, A; Vukušić, T; Stulić, V; Stanzer, D; Milošević, S

    2015-01-01

    The aim of this study was to investigate and compare two nonthermal techniques in the inactivation of moulds. High power ultrasound (20 kHz) and nonthermal gas phase plasma treatments were studied in the inactivation of selected moulds. Aspergillus spp. and Penicillium spp. were chosen as the most common mould present in or on food. Experimental design was introduced to establish and optimize working variables. For high power ultrasound, the greatest reduction of moulds (indicated by the total removal of viable cells) was obtained after ultrasound treatments at 60°C (thermosonication) for 6 and 9 min (power applied, 20-39 W). For plasma treatment, the greatest inactivation of moulds was observed for the longest treatment time (5 min) and lowest sample volume (2 ml), (AP12, AP13, PP12 and PP13). The great amount of applied energy required for achieving a partial log reduction in viable cells is the limiting factor for using high-power ultrasound. However, both treatment methods could be combined in the future to produce beneficial outcomes. This study deals with nonthermal food processing techniques and the results and findings present in this study are the root for further prospective studies. The food industry is looking for nonthermal methods that will enable food preservation, reduce deterioration of food compounds and structure and prolong food shelf life. © 2014 The Society for Applied Microbiology.

  11. The isolation and improvement of Aspergillus niger by radiation for higher production of citric acid

    International Nuclear Information System (INIS)

    Radziah A; Foziah Ali; Zainab H

    2000-01-01

    Local citric acid producer of fungal strain Aspergillus niger have been successfully isolated from stale bread and onion. The isolates, designated as SB 1 and NN I showed a potential performance for citric acid production of 49% and 52% yield respectively, in shake flask studies. The strain improvement on NN1 was carried out by radiation induced mutation by gamma rays at LD 5 0 of 1.28 kGy

  12. Karnatakafuran A and B: Two novel dibenzofurans isolated from the fungus Aspergillus karnatakaensis

    DEFF Research Database (Denmark)

    Manniche, S.; Sprogøe, K.; Dalsgaard, Petur

    2004-01-01

    Karnatakafurans A (1) and B (2), two novel dibenzofurans, have been isolated from the Specie Novum Aspergillus karnatakaensis Frisvad. The compounds were the major secondary metabolites and were isolated through UV-guided fractionation. of the organic extract. The structures were elucidated by sp...... by spectroscopic methods including MS and NMR. The compounds were tested for antimicrobial and antimalarial activity and proved to be moderately active against Plasmodium falciparum....

  13. A multiplex PCR method for detection of Aspergillus spp. and Mycobacterium tuberculosis in BAL specimens.

    Science.gov (United States)

    Amini, F; Kachuei, R; Noorbakhsh, F; Imani Fooladi, A A

    2015-06-01

    The aim of this study was the detection of Aspergillus species and Mycobacterium tuberculosis together in bronchoalveolar lavage (BAL) using of multiplex PCR. In this study, from September 2012 until June 2013, 100 bronchoalveolar lavage (BAL) specimens were collected from patients suspected of tuberculosis (TB). After the direct and culture test, multiplex PCR were utilized in order to diagnose Aspergillus species and M. tuberculosis. Phenol-chloroform manual method was used in order to extract DNA from these microorganisms. Aspergillus specific primers, M. tuberculosis designed primers and beta actin primers were used for multiplex PCR. In this study, by multiplex PCR method, Aspergillus species were identified in 12 samples (12%), positive samples in direct and culture test were respectively 11% and 10%. Sensitivity and specificity of this method in comparison to direct test were respectively 100% and 98.8%, also sensitivity and specificity of this method in comparison to culture test were respectively 100% and 97.7%. In this assay, M. tuberculosis was identified in 8 samples (8%). Mycobacterium-positive samples in molecular method, direct and culture test were respectively 6%, 5% and 7%. Sensitivity and specificity of PCR method in comparison to direct test were 80% and 97.8% also sensitivity and specificity of this method in comparison to culture test was 71.4% and 98.9%. In the present study, multiplex PCR method had higher sensitivity than direct and culture test in order to identify and detect Aspergillus, also this method had lower sensitivity for identification of M. tuberculosis, suggesting that the method of DNA extraction was not suitable. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  14. Resistance to Antibiotics in Strains of Staphylococcus spp., Enterococcus spp. and Escherichia coli Isolated from Rectal Swabs of Pigs

    Directory of Open Access Journals (Sweden)

    M. Kolář

    2008-01-01

    Full Text Available The study aimed at determining the level of resistance of selected bacterial species (Staphylococcus spp., Enterococcus spp., Escherichia coli isolated from rectal swabs of pigs to antimicrobial agents. The tested strains were isolated from piglets aged 7 to 30 days. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined quantitatively by the standard microdilution method. Resistance of the Staphylococcus aureus strain to oxacillin was confirmed by detection of the mecA gene and PBP2a. A total of 115 Staphylococcus spp. isolates were collected. In the case of Staphylococcus aureus, the methicillin-resistant strain (MRSA was identified. Moreover, higher frequency of coagulase-negative staphylococci with minimum inhibitory concentration of oxacillin ≥ 0.5 mg/l was noticed. Inducible resistance to clindamycin in the Staphylococcus hominis strain was also detected. The strains of Enterococcus spp. (61 isolates exhibited high resistance to tetracycline (98.5%, erythromycin (86.8% and chloramphenicol (54.4%. Vancomycin-resistant enterococci were not isolated. In the case of Escherichia coli strains (111 isolates, higher frequency of resistant strains to tetracycline (81.1% and ampicillin (62.2% was documented. Resistance to fluoroquinolones and production of broad-spectrum β-lactamases was not noticed. The presented study may be considered as a pilot project assessing the prevalence of resistant bacteria in piglets kept on a single farm. It demonstrated the presence of resistant strains of Staphylococcus spp., including one MRSA strain, Enterococcus spp. and Escherichia coli. These strains may be present as a result of postnatal colonization with both bacterial microflora of dams and environmental microflora.

  15. PROBIOTIC POTENCY OF LACTOBACILLUS SPP. ISOLATED FROM SUMBAWA MARE MILK

    Directory of Open Access Journals (Sweden)

    I Nengah Sujaya

    2008-03-01

    Full Text Available This research was deigned to elucidate the potency of Lactobacillus spp. isolated from sumbawa mare milk to be developed as a probiotic. Sixteen lacobacilli were screened based on their resitancy to a model of gastric juice at pH 2, 3, and 4, then followed by their resistncy to small intestional fluid model containing deoxycholic. Three lactobacilli i.e. Lactobacillus sp. SKA13, Lactobacillus rhamnosus SKG34 and Lactobacillus rhamnosus SKG49 were found to be resistentent to gastric juice at pH 3 and 4. However, there were no lactobacilli resisted to pH 2. Lactobacillus rhamnosus SKG34 and Lactobacillus rhamnosus SKG49 were able to reach the colon even after being expossed to a model of intestinal fluid containing 0,4 mM deoxycholate and pancreatine. Therefore, these isolates have a potency to be developed as probiotic lactobacilli. Nevertherless, these lactobcailli could probably transform cholic acid into secondary bile acids, which were not expected to be found in the probiotic, and this capability is not appropriate for probiotic. This character is worthly to be studied since it has never been reported in lactobacilli.

  16. Pathogenicity of isolates of Colletotrichum spp.: The causal agents of anthracnose

    OpenAIRE

    Živković, Svetlana; Dolovac, Nenad; Popović, Tatjana; Stojanović, Saša

    2012-01-01

    The pathogenic characteristics of 20 isolates of Colletotrichum spp. originating from pear, apple, sour cherry and tomato fruits, as well as reference strains of C. acutatum (CBS 294.67) and C. gloeosporioides (CBS 516.97) are presented in this paper. In the studies of host range of isolates of Colletotrichum spp. were included 17 plant species. Nine days after artificial inoculation all tested isolates were caused anthracnose lesion on fruits of apple, pear, peach, apricot, sour cherry, swee...

  17. Molecular characterisation of Aspergillus flavus isolates from peanut fields in India using AFLP

    Directory of Open Access Journals (Sweden)

    Diwakar Singh

    2015-09-01

    Full Text Available Aflatoxin contamination of peanut, due to infection by Aspergillus flavus, is a major problem of rain-fed agriculture in India. In the present study, molecular characterisation of 187 Aspergillus flavus isolates, which were sampled from the peanut fields of Gujarat state in India, was performed using AFLP markers. On a pooled cluster analysis, the markers could successfully discriminate among the ‘A’, ‘B’ and ‘G’ group A. flavus isolates. PCoA analysis also showed equivalent results to the cluster analysis. Most of the isolates from one district could be clustered together, which indicated genetic similarity among the isolates. Further, a lot of genetic variability was observed within a district and within a group. The results of AMOVA test revealed that the variance within a population (84% was more than that between two populations (16%. The isolates, when tested by indirect competitive ELISA, showed about 68.5% of them to be atoxigenic. Composite analysis between the aflatoxin production and AFLP data was found to be ineffective in separating the isolate types by aflatoxigenicity. Certain unique fragments, with respect to individual isolates, were also identified that may be used for development of SCAR marker to aid in rapid and precise identification of isolates.

  18. Molecular identification and amphotericin B susceptibility testing of clinical isolates of Aspergillus from 11 hospitals in Korea.

    Science.gov (United States)

    Heo, Min Seok; Shin, Jong Hee; Choi, Min Ji; Park, Yeon Joon; Lee, Hye Soo; Koo, Sun Hoe; Lee, Won Gil; Kim, Soo Hyun; Shin, Myung Geun; Suh, Soon Pal; Ryang, Dong Wook

    2015-11-01

    We investigated the species distribution and amphotericin B (AMB) susceptibility of Korean clinical Aspergillus isolates by using two Etests and the CLSI broth microdilution method. A total of 136 Aspergillus isolates obtained from 11 university hospitals were identified by sequencing the internal transcribed spacer (ITS) and β-tubulin genomic regions. Minimal inhibitory concentrations (MICs) of AMB were determined in Etests using Mueller-Hinton agar (Etest-MH) and RPMI agar (Etest-RPG), and categorical agreement with the CLSI method was assessed by using epidemiological cutoff values. ITS sequencing identified the following six Aspergillus species complexes: Aspergillus fumigatus (42.6% of the isolates), A. niger (23.5%), A. flavus (17.6%), A. terreus (11.0%), A. versicolor (4.4%), and A. ustus (0.7%). Cryptic species identifiable by β-tubulin sequencing accounted for 25.7% (35/136) of the isolates. Of all 136 isolates, 36 (26.5%) had AMB MICs of ≥2 μg/mL by the CLSI method. The categorical agreement of Etest-RPG with the CLSI method was 98% for the A. fumigatus, A. niger, and A. versicolor complexes, 87% for the A. terreus complex, and 37.5% for the A. flavus complex. That of Etest-MH was ≤75% for the A. niger, A. flavus, A. terreus, and A. versicolor complexes but was higher for the A. fumigatus complex (98.3%). Aspergillus species other than A. fumigatus constitute about 60% of clinical Aspergillus isolates, and reduced AMB susceptibility is common among clinical isolates of Aspergillus in Korea. Molecular identification and AMB susceptibility testing by Etest-RPG may be useful for characterizing Aspergillus isolates of clinical relevance.

  19. Performance of Matrix-Assisted Laser Desorption Ionization−Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting

    Science.gov (United States)

    Sleiman, Sue; Halliday, Catriona L.; Chapman, Belinda; Brown, Mitchell; Nitschke, Joanne; Lau, Anna F.

    2016-01-01

    We developed an Australian database for the identification of Aspergillus, Scedosporium, and Fusarium species (n = 28) by matrix-assisted laser desorption ionization−time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%; Fusarium, 84% versus 42%; and Scedosporium, 94% versus 18%, respectively). PMID:27252460

  20. Occurrence and antimicrobial resistance of Salmonella spp. isolated from food other than meat in Poland

    Directory of Open Access Journals (Sweden)

    Łukasz Mąka

    2015-09-01

    Full Text Available Introduction and objectives. Antimicrobial resistance of pathogenic bacteria can result in therapy failure, increased hospitalization, and increased risk of death. In Poland, [i]Salmonella[/i] spp. is a major bacterial agent of food poisoning. The majority of studies on antimicrobial resistance in [i]Salmonella[/i] spp. isolates from food have focused on meat products as the source of this pathogen. In comparison, this study examines the antimicrobial susceptibility of [i]Salmonella[/i] spp. isolated from retail food products other than meat in Poland. Materials and Methods. A collection of 122 [i]Salmonella[/i] spp. isolates were isolated in Poland in 2008–2012 from foods other than meat: confectionery products, eggs, fruits, vegetables, spices and others. The resistance of these isolates to 19 antimicrobial agents was tested using the disc diffusion method. Results. [i]Salmonella[/i] Enteritidis was the most frequently identified serotype (84.4% of all tested isolates. In total, 42.6% of the [i]Salmonella[/i] spp. isolates were resistant to antibiotics. The highest frequencies of resistance were observed in isolates from 2009 (60.0% and 2012 (59.5%. Antibiotic resistance was most prevalent among [i]Salmonella[/i] spp. isolated from egg-containing food samples (68.0%. Resistance to nalidixic acid was most common and was observed in 35.2% of all tested isolates. The isolates were less frequently resistant to sulphonamides (6.6%, ampicillin (4.9%, amoxicillin/clavulanic acid (2.5% and to streptomycin, cefoxitin, gentamicin and tetracycline (1.6%. Only one isolate showed resistance to chloramphenicol. Four isolates displayed multiresistance. Conclusions. Although, the level of resistance and multiresistance of [i]Salmonella[/i] spp. isolates from non-meat foods was lower than in those from meat products, the presence of these resistant bacteria poses a real threat to the health of consumers.

  1. [The isolation and evaluation of Aspergillus fumigatus antigens].

    Science.gov (United States)

    Lirio, V de S; de Assis, C M; Cano, M I; Lacaz, C da S

    1992-01-01

    Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electrophoretic profile (26 proteic subunits with a molecular weight ranging from 18 a > 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immunoblotting against the antiserum was carried out.

  2. The isolated Leptospira Spp. Identification by molecular biological techniques

    Directory of Open Access Journals (Sweden)

    Duangjai Suwancharoen

    2017-01-01

    Full Text Available Leptospirosis is a zoonotic disease caused by the bacteria of Leptospira spp. Identification of this bacterium relies on serotyping and genotyping. Data base for animal causative serovars in Thailand is limited. As the unknown serovars are found in the laboratory, they need to be sent overseas for referent identification. To reduce the cost, this research intended to develop a leptospiral identification method which is user–friendly and able to classify efficiently. Ten Leptospira isolations were cultured from urine samples. They were identified by three molecular biological techniques, including Pulsed-Field Gel Electrophoresis (PFGE, Variable Number Tandem Repeat (VNTR and Multilocus Sequence Typing (MLST. These methods were developed and compared to find the most suitable one for leptospiral identification. VNTR was found to be inappropriate since it could not identify the agents and it did not show the PCR product. PFGE and MLST gave the same results of the unknown 1 and 2 which were L.weilii sv Samin st Samin. Unknown 4 showed different results by each technique. Unknown 5 to 10 were likely to be L.meyeri sv Ranarum st ICF and Leptonema illini sv Illini st 3055 by PFGE but MLST could not identify the serovar. However, molecular biological technique for Leptospira identification should be done by several methods in order to confirm the result of each other.

  3. Brucella papionis sp. nov., isolated from baboons (Papio spp.).

    Science.gov (United States)

    Whatmore, Adrian M; Davison, Nicholas; Cloeckaert, Axel; Al Dahouk, Sascha; Zygmunt, Michel S; Brew, Simon D; Perrett, Lorraine L; Koylass, Mark S; Vergnaud, Gilles; Quance, Christine; Scholz, Holger C; Dick, Edward J; Hubbard, Gene; Schlabritz-Loutsevitch, Natalia E

    2014-12-01

    Two Gram-negative, non-motile, non-spore-forming coccoid bacteria (strains F8/08-60(T) and F8/08-61) isolated from clinical specimens obtained from baboons (Papio spp.) that had delivered stillborn offspring were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence similarities, both strains, which possessed identical sequences, were assigned to the genus Brucella. This placement was confirmed by extended multilocus sequence analysis (MLSA), where both strains possessed identical sequences, and whole-genome sequencing of a representative isolate. All of the above analyses suggested that the two strains represent a novel lineage within the genus Brucella. The strains also possessed a unique profile when subjected to the phenotyping approach classically used to separate species of the genus Brucella, reacting only with Brucella A monospecific antiserum, being sensitive to the dyes thionin and fuchsin, being lysed by bacteriophage Wb, Bk2 and Fi phage at routine test dilution (RTD) but only partially sensitive to bacteriophage Tb, and with no requirement for CO2 and no production of H2S but strong urease activity. Biochemical profiling revealed a pattern of enzyme activity and metabolic capabilities distinct from existing species of the genus Brucella. Molecular analysis of the omp2 locus genes showed that both strains had a novel combination of two highly similar omp2b gene copies. The two strains shared a unique fingerprint profile of the multiple-copy Brucella-specific element IS711. Like MLSA, a multilocus variable number of tandem repeat analysis (MLVA) showed that the isolates clustered together very closely, but represent a distinct group within the genus Brucella. Isolates F8/08-60(T) and F8/08-61 could be distinguished clearly from all known species of the genus Brucella and their biovars by both phenotypic and molecular properties. Therefore, by applying the species concept for the genus Brucella suggested by the ICSP

  4. Alicyclobacillus spp. in the fruit juice industry: history, characteristics, and current isolation/detection procedures.

    Science.gov (United States)

    Chang, Su-Sen; Kang, Dong-Hyun

    2004-01-01

    The first Alicyclobacillus spp. was isolated in 1982, and was originally thought to be strictly limited to thermophilic and acidic environments. Two years later, another Alicyclobacillus sp., A. acidoterrestris, was identified as the causative agent in spoilage of commercially pasteurized apple juice. Subsequent studies soon found that Alicyclobacillus spp. are soilborne bacteria, and do not strictly require thermophilic and acidic environments. Alicyclobacillus spp. posess several distinct characteristics; the major one is their ability to survive commercial pasteurization processes and produce off-flavors in fruit juices. The fruit juice industry has acknowledged Alicyclobacillus spp. as a major quality control target microorganism. Guaiacol and halophenols were identified as the offensive smelling agent in many Alicyclobacillus spp. related spoilage. Though the exact formation pathway of these off-flavors by Alicyclobacillus spp. are not yet identified, studies report that the presence of Alicyclobacillus spp. in the medium may be a major contributor to the formation of these off-flavors. Many identification methods and isolation media were developed in the last two decades. However, most of these methods were developed specifically for A. acidoterrestris, which was the first identified off-flavor producing Alicyclobacillus. However, recent studies indicate that other species of Alicyclobacillus may also produce guaiacol or the halophenols. In this respect, all Alicyclobacillus spp. should be monitored as potential spoilage bacteria in fruit juices. This article includes an overall review of the history of Alicyclobacillus spp., characteristics, suggested off-flavor production pathways, and commonly used identification methods for the currently identified Alicyclobacillus spp.

  5. Extracellular Xylanolytic and Pectinolytic Hydrolase Production by Aspergillus flavus Isolates Contributes to Crop Invasion

    Directory of Open Access Journals (Sweden)

    Jay E. Mellon

    2015-08-01

    Full Text Available Several atoxigenic Aspergillus flavus isolates, including some being used as biocontrol agents, and one toxigenic isolate were surveyed for the ability to produce extracellular xylanolytic and pectinolytic hydrolases. All of the tested isolates displayed good production of endoxylanases when grown on a medium utilizing larch xylan as a sole carbon substrate. Four of the tested isolates produced reasonably high levels of esterase activity, while the atoxigenic biocontrol agent NRRL 21882 isolate esterase level was significantly lower than the others. Atoxigenic A. flavus isolates 19, 22, K49, AF36 (the latter two are biocontrol agents and toxigenic AF13 produced copious levels of pectinolytic activity when grown on a pectin medium. The pectinolytic activity levels of the atoxigenic A. flavus 17 and NRRL 21882 isolates were significantly lower than the other tested isolates. In addition, A. flavus isolates that displayed high levels of pectinolytic activity in the plate assay produced high levels of endopolygalacturonase (pectinase P2c, as ascertained by isoelectric focusing electrophoresis. Isolate NRRL 21882 displayed low levels of both pectinase P2c and pectin methyl esterase. A. flavus appears capable of producing these hydrolytic enzymes irrespective of aflatoxin production. This ability of atoxigenic isolates to produce xylanolytic and pectinolytic hydrolases mimics that of toxigenic isolates and, therefore, contributes to the ability of atoxigenic isolates to occupy the same niche as A. flavus toxigenic isolates.

  6. Antimicrobial resistance profile of Enterococcus spp isolated from food in Southern Brazil

    Science.gov (United States)

    Riboldi, Gustavo Pelicioli; Frazzon, Jeverson; d’Azevedo, Pedro Alves; Frazzon, Ana Paula Guedes

    2009-01-01

    Fifty-six Enterococcus spp. strains were isolated from foods in Southern Brazil, confirmed by PCR and classified as Enterococcus faecalis (27), Enterococcus faecium (23) and Enterococcus spp (6). Antimicrobial susceptibility tests showed resistance phenotypes to a range of antibiotics widely administrated in humans such as gentamycin, streptomycin, ampicillin and vancomycin. PMID:24031330

  7. Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee.

    Science.gov (United States)

    von Hertwig, Aline Morgan; Sant'Ana, Anderson S; Sartori, Daniele; da Silva, Josué José; Nascimento, Maristela S; Iamanaka, Beatriz Thie; Pelegrinelli Fungaro, Maria Helena; Taniwaki, Marta Hiromi

    2018-05-01

    Some species from Aspergillus section Nigri are morphologically very similar and altogether have been called A. niger aggregate. Although the species included in this group are morphologically very similar, they differ in their ability to produce mycotoxins and other metabolites and their taxonomical status has evolved continuously. Among them, A. niger and A. welwitschiae are ochratoxin A and fumonisin B 2 producers and their detection and/or identification is of crucial importance for food safety. The aim of this study was the development of a real-time PCR-based method for simultaneous discrimination of A. niger and A. welwitschiae from other species of the A. niger aggregate isolated from coffee beans. One primer pair and a hybridization probe specific for detection of A. niger and A. welwitschiae strains were designed based on the BenA gene sequences, and used in a Real-time PCR assay for the rapid discrimination between both these species from all others of the A. niger aggregate. The Real-time PCR assay was shown to be 100% efficient in discriminating the 73 isolates of A. niger/A. welwitschiae from the other A. niger aggregate species analyzed as a negative control. This result testifies to the use of this technique as a good tool in the rapid detection of these important toxigenic species. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Aspergillus spp. colonization in exhaled breath condensate of lung cancer patients from Puglia Region of Italy.

    Science.gov (United States)

    Carpagnano, Giovanna E; Lacedonia, Donato; Palladino, Grazia Pia; Logrieco, Giuseppe; Crisetti, Elisabetta; Susca, Antonia; Logrieco, Antonio; Foschino-Barbaro, Maria P

    2014-02-18

    Airways of lung cancer patients are often colonized by fungi. Some of these colonizing fungi, under particular conditions, produce cancerogenic mycotoxins. Given the recent interest in the infective origin of lung cancer, with this preliminary study we aim to give our small contribution to this field of research by analysing the fungal microbiome of the exhaled breath condensate of lung cancer patients from Puglia, a region of Italy. We enrolled 43 lung cancer patients and 21 healthy subjects that underwent exhaled breath condensate and bronchial brushing collection. The fungal incidence and nature of sample collected were analysed by using a selected media for Aspergillus species. For the first time we were able to analyse the fungal microbioma of the exhaled breath condensate. 27.9% of lung cancer patients showed a presence of Aspergillus niger, or A. ochraceus or Penicillium ssp. while none of the healthy subjects did so. The results confirmed the high percentage of fungal colonization of the airways of lung cancer patients from Puglia, suggesting the need to conduct further analyses in this field in order to evaluate the exact pathogenetic role of these fungi in lung cancer as well as to propose efficient, empirical therapy.

  9. In vitro susceptibility of 188 clinical and environmental isolates of Aspergillus flavus for the new triazole isavuconazole and seven other antifungal drugs

    NARCIS (Netherlands)

    Shivaprakash, M.R.; Geertsen, E.; Chakrabarti, A.; Mouton, J.W.; Meis, J.F.G.M.

    2011-01-01

    Recently isavuconazole, an experimental triazole agent, was found to be active against Aspergillus species. As Aspergillus flavus is the second-most common Aspergillus species isolated from human infection and the fungus has not been widely tested against the drug, we studied a large collection of

  10. Occurrence and antimicrobial resistance of Salmonella spp. isolated from food other than meat in Poland

    Directory of Open Access Journals (Sweden)

    Łukasz Mąka

    2015-09-01

    Although, the level of resistance and multiresistance of Salmonella spp. isolates from non-meat foods was lower than in those from meat products, the presence of these resistant bacteria poses a real threat to the health of consumers.

  11. Isolation, characterization and optimization of culture parameters for production of an alkaline protease isolated from Aspergillus tamarii.

    Science.gov (United States)

    Anandan, Dayanandan; Marmer, William N; Dudley, Robert L

    2007-05-01

    Aspergillus tamarii expresses an extracellular alkaline protease that we show to be effective in removing hair from cattle hide. Large quantities of the enzyme will be required for the optimization of the enzymatic dehairing process so the growth conditions for maximum protease expression by A. tamarii were optimized for both solid-state culture on wheat bran and for broth culture. Optimal protease expression occurred, for both cultural media, at initial pH 9; the culture was incubated at 30 degrees C for 96 h using a 5% inoculum. The crude enzyme was isolated, purified and characterized using MALDI TOF TOF. The alkaline protease was homologous to the alkaline protease expressed by Aspergillus viridinutans.

  12. [Isolation of Aspergillus tritici from internal environment (Chile): Ecological and clinical scope].

    Science.gov (United States)

    Vieille Oyarzo, Peggy; Cruz Choappa, Rodrigo; Piontelli Laforet, Eduardo

    2018-03-29

    Indoor environments provide important protective habitats for humans, who live or work in them most of the time. Many of these environments lack ventilation, which affects the composition of microbial communities, especially that of the fungal community. The aim of this study is to report the isolation of Aspergillus section Candidi from indoor environments of the School of Medicine at Universidad de Valparaiso, Chile, and identification through morpho-physiological and molecular approaches. Their ecological and clinical features were highlighted. An environmental non-volumetric sampling was performed on PDA medium; 2 petri dishes were exposed in 10 different places to select the Aspergillus samples. Subcultures were performed on agar Czapek with yeast extract (CYA), malt extract agar (MEA) and creatin sacarose agar (CREA) media only for the morpho-physiological and later the molecular identification of white spore species. Of the 20 samples analyzed, one Aspergillus belonging to Candidi section was isolated. Based on its morphology and molecular features, it was classified as Aspergillustritici Mehrotra & Basu. Its ecology and medical relevance are reviewed and discussed. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Bacillus Spp. isolated from the conjunctiva and their potential ...

    African Journals Online (AJOL)

    2014-06-02

    Jun 2, 2014 ... Introduction. Application of antibiotics in the treatment of bacterial ... Keywords: Bacillus spp, antibacterial activity, eyes pathogens, conjunctiva. African Health ... ml of respective test organism and allowed to dry. In the agar ...

  14. Molecular characterization of Azotobacter spp. nifH gene Isolated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-15

    Dec 15, 2009 ... Available online at http://www.academicjournals.org/AJB. ISSN 1684–5315 ... Molecular characterization of Azotobacter spp. nifH .... MATERIALS AND METHODS ..... rapidly expanding and is currently composed of over.

  15. Conventional Morphology Versus PCR Sequencing, rep-PCR, and MALDI-TOF-MS for Identification of Clinical Aspergillus Isolates Collected Over a 2-Year Period in a University Hospital at Kayseri, Turkey.

    Science.gov (United States)

    Atalay, Altay; Koc, Ayse Nedret; Suel, Ahmet; Sav, Hafize; Demir, Gonca; Elmali, Ferhan; Cakir, Nuri; Seyedmousavi, Seyedmojtaba

    2016-09-01

    Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains. A total of 24 consecutive clinical isolates of Aspergillus were collected during 2012-2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara. Samples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001). The good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required. © 2016 Wiley Periodicals, Inc.

  16. Isolate-dependent growth, virulence, and cell wall composition in the human pathogen Aspergillus fumigatus.

    Directory of Open Access Journals (Sweden)

    Nansalmaa Amarsaikhan

    Full Text Available The ubiquitous fungal pathogen Aspergillus fumigatus is a mediator of allergic sensitization and invasive disease in susceptible individuals. The significant genetic and phenotypic variability between and among clinical and environmental isolates are important considerations in host-pathogen studies of A. fumigatus-mediated disease. We observed decreased radial growth, rate of germination, and ability to establish colony growth in a single environmental isolate of A. fumigatus, Af5517, when compared to other clinical and environmental isolates. Af5517 also exhibited increased hyphal diameter and cell wall β-glucan and chitin content, with chitin most significantly increased. Morbidity, mortality, lung fungal burden, and tissue pathology were decreased in neutropenic Af5517-infected mice when compared to the clinical isolate Af293. Our results support previous findings that suggest a correlation between in vitro growth rates and in vivo virulence, and we propose that changes in cell wall composition may contribute to this phenotype.

  17. Isolation and chemical characterization of naphthoquinone metabolites of Aspergillus parvulus Smith

    International Nuclear Information System (INIS)

    Wang, C.C.P.

    1984-01-01

    Although several benzoquinone and anthraquinone compounds have been isolated from Aspergillus species, only two naphthoquinone monomers have been reported thus far. Aspergillus parvulus Smith (ATCC number16911) was first investigated chemically in 1974, and five naphthalenones, along with one naphthoquinone, were isolated and characterized. Based on biosynthetic considerations, it was thought that A. parvulus might be capable of producing additional naphthoquinones under suitable conditions. It was decided to undertake a further investigation of A. parvulus. Thus, three novel naphthoquinones, compounds A, B, and C, were isolated from A. parvulus cultures grown in an acidic medium of glucose and phytone peptone. The structures of these compounds were deduced largely by the comparison of the effects of acetylation on the 1 H-NMR and 13 C-NMR spectra of the parent compounds and their four derivatives. An unusual mass fragmentation pattern which was previously thought to be unfavorable was discovered, and the other fragmentation patterns of the parent compounds, as well as their derivatives, were proposed. This investigation appears to be the third reported isolation of 2,5,7-tri-hydroxy-1,4-naphthoquinone derivatives from nature and the first reported from A. parvulus

  18. Sarcocystis neurona and Neospora caninum in Brazilian opossums (Didelphis spp.): Molecular investigation and in vitro isolation of Sarcocystis spp.

    Science.gov (United States)

    Gondim, Leane S Q; Jesus, Rogério F; Ribeiro-Andrade, Müller; Silva, Jean C R; Siqueira, Daniel B; Marvulo, Maria F V; Aléssio, Felipe M; Mauffrey, Jean-François; Julião, Fred S; Savani, Elisa San Martin Mouriz; Soares, Rodrigo M; Gondim, Luís F P

    2017-08-30

    Sarcocystis neurona and Neospora spp. are protozoan parasites that induce neurological diseases in horses and other animal species. Opossums (Didelphis albiventris and Didelphis virginiana) are definitive hosts of S. neurona, which is the major cause of equine protozoal myeloencephalitis (EPM). Neospora caninum causes abortion in cattle and infects a wide range of animal species, while N. hughesi is known to induce neurologic disease in equids. The aims of this study were to investigate S. neurona and N. caninum in tissues from opossums in the northeastern Brazil, and to isolate Brazilian strains of Sarcocystis spp. from wild opossums for comparison with previously isolated strains. Carcasses of 39 opossums from Bahia state were available for molecular identification of Sarcocystis spp. and N. caninum in their tissues, and for sporocyst detection by intestinal scraping. In addition, Sarcocystis-like sporocysts from nine additional opossums, obtained in São Paulo state, were tested. Sarcocystis DNA was found in 16 (41%) of the 39 opossums' carcasses; N. caninum DNA was detected in tissues from three opossums. The sporocysts from the nine additional opossums from São Paulo state were tested by bioassay and induced infection in nine budgerigars, but in none of the gamma-interferon knockout mice. In vitro isolation was successful using tissues from all nine budgerigars. The isolated strains were maintained in CV-1 and Vero cells. Three of nine isolates presented contamination in cell culture and were discarded. Analysis of six isolates based on five loci showed that these parasites were genetically different from each other and also distinct from S. neurona, S. falcatula, S. lindsayi, and S. speeri. In conclusion, opossums in the studied regions were infected with N. caninum and Sarcocystis spp. and represent a potential source of infection to other animals. This is the first report of N. caninum infection in tissues from black-eared opossum (D. aurita or D

  19. Species identification of Aspergillus section Flavi isolates from Portuguese almonds using phenotypic, including MALDI-TOF ICMS, and molecular approaches.

    Science.gov (United States)

    Rodrigues, P; Santos, C; Venâncio, A; Lima, N

    2011-10-01

    Section Flavi is one of the most significant sections in the genus Aspergillus. Taxonomy of this section currently depends on multivariate approaches, entailing phenotypic and molecular traits. This work aimed to identify isolates from section Flavi by combining various classic phenotypic and genotypic methods as well as the novel approach based on spectral analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF ICMS) and to evaluate the discriminatory power of the various approaches in species identification.   Aspergillus section Flavi isolates obtained from Portuguese almonds were characterized in terms of macro- and micromorphology, mycotoxin pattern, calmodulin gene sequence and MALDI-TOF protein fingerprint spectra. For each approach, dendrograms were created and results were compared. All data sets divided the isolates into three groups, corresponding to taxa closely related to Aspergillus flavus, Aspergillus parasiticus and Aspergillus tamarii. In the A. flavus clade, molecular and spectral analyses were not able to resolve between aflatoxigenic and nonaflatoxigenic isolates. In the A. parasiticus cluster, two well-resolved clades corresponded to unidentified taxa, corresponding to those isolates with mycotoxin profile different from that expected for A. parasiticus. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  20. Aspergillus species as emerging causative agents of onychomycosis.

    Science.gov (United States)

    Nouripour-Sisakht, S; Mirhendi, H; Shidfar, M R; Ahmadi, B; Rezaei-Matehkolaei, A; Geramishoar, M; Zarei, F; Jalalizand, N

    2015-06-01

    Onychomycosis is a common nail infection caused by dermatophytes, non-dermatophyte molds (NDM), and yeasts. Aspergillus species are emerging as increasing causes of toenail onychomycosis. The purpose of this study was species delineation of Aspergillus spp. isolated from patients with onychomycosis. During a period of one year (2012-2013), nail samples were collected from patients clinically suspected of onychomycosis and subjected to microscopic examination and culture. Species identification was performed based on macro- and micro-morphology of colonies. For precise species identification, PCR-amplification and sequencing of the beta-tubulin gene followed by BLAST queries were performed where required. A total of 463/2,292 (20.2%) tested nails were diagnosed with onychomycosis. Among the positive specimens, 154 cases (33.2%) were identified as saprophytic NDM onychomycosis, 135 (29.2%) of which were attributable to Aspergillus. Aspergillus species isolated from the infected nails included Aspergillus flavus (77.3%, n=119), Aspergillus niger (n=4), Aspergillus tubingensis (n=4), Aspergillus terreus (n=3), Aspergillus sydowii (n=2), Aspergillus spp. (n=2), and Aspergillus candidus (n=1). Among the patients diagnosed with onychomycosis due to Aspergillus (average patient age, 47.4 years), 40 had fingernail and 95 toenail involvement. The large toenails were most commonly affected. This study identified a markedly high occurrence of A. flavus, and this fungus appears to be an emerging cause of saprophytic onychomycosis in Iran. The study moreover highlights the necessity of differentiating between dermatophytic and non-dermatophytic nail infections for informed decisions on appropriate therapy. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  1. Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum trin

    Energy Technology Data Exchange (ETDEWEB)

    Correa, Marivaldo J.C.; Nunes, Fatima M.; Bitencourt, Heriberto R.; Borges, Fabio C.; Guilhon, Giselle M.S.P.; Arruda, Mara S.P.; Marinho, Andrey M. R.; Santos, Alberdan S.; Alves, Claudio N.; Santos, Lourivaldo S., E-mail: lss@ufpa.b [Universidade Federal do Para (IQ/FEQ/UFPA), Belem, PA (Brazil). Inst. de Tecnologia. Faculdade de Engenharia Quimica; Brasil, Davi S.B. [Universidade Federal do Para (PPGQ/IQ/UFPA), Belem, PA (Brazil). Inst. de Quimica. Programa de Pos-Graduacao em Quimica

    2011-07-01

    The fungus Aspergillus flavus isolated as endophytic of the plant Paspalum maritimum Trin. was evaluated for its potential application in biotransformation reactions. The compounds chalcone (1), 3,4,5-trimethoxychalcone (2) and 2,3,4,4'-tetramethoxy chalcone (3) were biotransformed, respectively, in dihydrochalcone (4), 3,4,5-trimethoxydihydrochalcone (5) and 2,3,4,4'-tetramethoxydihydrochalcone (6). The structures were elucidated by spectroscopic methods including 1D and 2D NMR techniques, and MS analysis. The dihydrochalcones 5 and 6 are new compounds. (author)

  2. Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum trin

    International Nuclear Information System (INIS)

    Correa, Marivaldo J.C.; Nunes, Fatima M.; Bitencourt, Heriberto R.; Borges, Fabio C.; Guilhon, Giselle M.S.P.; Arruda, Mara S.P.; Marinho, Andrey M. R.; Santos, Alberdan S.; Alves, Claudio N.; Santos, Lourivaldo S.; Brasil, Davi S.B.

    2011-01-01

    The fungus Aspergillus flavus isolated as endophytic of the plant Paspalum maritimum Trin. was evaluated for its potential application in biotransformation reactions. The compounds chalcone (1), 3,4,5-trimethoxychalcone (2) and 2,3,4,4'-tetramethoxy chalcone (3) were biotransformed, respectively, in dihydrochalcone (4), 3,4,5-trimethoxydihydrochalcone (5) and 2,3,4,4'-tetramethoxydihydrochalcone (6). The structures were elucidated by spectroscopic methods including 1D and 2D NMR techniques, and MS analysis. The dihydrochalcones 5 and 6 are new compounds. (author)

  3. Frequency, virulence genes and antimicrobial resistance of Listeria spp. isolated from bovine clinical mastitis.

    Science.gov (United States)

    Jamali, Hossein; Radmehr, Behrad

    2013-11-01

    The aims of this study were to determine the prevalence, characteristics and antimicrobial resistance of Listeria spp. isolated from bovine clinical mastitis in Iran. Listeria spp. were detected in 21/207 bovine mastitic milk samples from dairy farms in Iran, comprising L. monocytogenes (n=17), L. innocua (n=3) and L. ivanovii (n=1). L. monocytogenes isolates were grouped into serogroups '4b, 4d, 4e', '1/2a, 3a', '1/2b, 3b, 7' and '1/2c, 3c'; all harboured inlA, inlC and inlJ virulence genes. Listeria spp. were most frequently resistant to penicillin G (14/21 isolates, 66.7%) and tetracyclines (11/21 isolates, 52.4%). Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Isolation and molecular identification of Vibrio spp. by sequencing of ...

    African Journals Online (AJOL)

    Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6% of samples of meat and meat products showed colonies on TCBS.

  5. Isolation, Identification, and Characterization of a New Highly Pathogenic Field Isolate of Mycobacterium avium spp. avium

    Directory of Open Access Journals (Sweden)

    Liangquan Zhu

    2018-01-01

    Full Text Available Avian tuberculosis is a chronic, contagious zoonotic disease affecting birds, mammals, and humans. The disease is most often caused by Mycobacterium avium spp. avium (MAA. Strain resources are important for research on avian tuberculosis and vaccine development. However, there has been little reported about the newly identified MAA strain in recent years in China. In this study, a new strain was isolated from a fowl with symptoms of avian tuberculosis by bacterial culture. The isolated strain was identified to be MAA by culture, staining, and biochemical and genetic analysis, except for different colony morphology. The isolated strain was Ziehl-Zeelsen staining positive, resistant to p-nitrobenzoic acid, and negative for niacin production, Tween-80 hydrolysis, heat stable catalase and nitrate production. The strain had the DnaJ gene, IS1245, and IS901, as well. Serum agglutination indicated that the MAA strain was of serotype 1. The MAA strain showed strong virulence via mortality in rabbits and chickens. The prepared tuberculin of the MAA strain had similar potency compared to the MAA reference strain and standard tuberculin via a tuberculin skin test. Our studies suggested that this MAA strain tends to be a novel subtype, which might enrich the strain resource of avian tuberculosis.

  6. Isolation and Determination of Antibiotic Resistance Patterns in Nontyphoid Salmonella spp isolated from chicken

    Directory of Open Access Journals (Sweden)

    Seyyedeh Hoorieh Fallah

    2013-01-01

    Full Text Available Background: Salmonellosis is one of the most common food borne diseases in industrial and developing countries. In recent years, an increase in antimicrobial drug resistance, among non-typhoid Salmonella spp has been observed. Objectives: The aim of this study was to isolate and determine antibiotic resistance pattern in non-typhoid Salmonella spp. Materials and Methods: This descriptive study was done on 100 samples of chickens collected from 196 retail markets and was examined for the presence of Salmonella using standard bacteriological procedures and stereotyping kit. Antimicrobial susceptibility testing was performed by disk diffusion methods according to the National Committee for Clinical Laboratory Standards (CLSI. The data were analyzed by using the SPSS software version 18. Result: Forty- four percent of samples were contaminated with Salmonella infection and 56% didn’t have any contamination. The stereotyping results showed that 34 of 44 isolates of Salmonella belonged to Salmonella infantis (79.5 %, one strain (2.3% of group C and 8 strain (18.2% of group D. However, all these strains were sensitive to Cefotaxime and Ciprofloxacin, and 100% were resistant to Nalidixic acid, Tetracyclin and Sterptomycin. The most common resistance pattern (34.1% was towards six antibiotics, and 6.8% of strains were resistant to at least three antibiotics. Conclusion: High levels of resistance to antibiotics that are used commonly for human and poultry can be a warning for our community health and this information must be used to form important strategies for improvement of infection control.

  7. Performance of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Aspergillus, Scedosporium, and Fusarium spp. in the Australian Clinical Setting.

    Science.gov (United States)

    Sleiman, Sue; Halliday, Catriona L; Chapman, Belinda; Brown, Mitchell; Nitschke, Joanne; Lau, Anna F; Chen, Sharon C-A

    2016-08-01

    We developed an Australian database for the identification of Aspergillus, Scedosporium, and Fusarium species (n = 28) by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In a challenge against 117 isolates, species identification significantly improved when the in-house-built database was combined with the Bruker Filamentous Fungi Library compared with that for the Bruker library alone (Aspergillus, 93% versus 69%; Fusarium, 84% versus 42%; and Scedosporium, 94% versus 18%, respectively). Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  8. Isolation, characterization and transcriptome analysis of a novel Antarctic Aspergillus sydowii strain MS-19 as a potential lignocellulosic enzyme source.

    Science.gov (United States)

    Cong, Bailin; Wang, Nengfei; Liu, Shenghao; Liu, Feng; Yin, Xiaofei; Shen, Jihong

    2017-05-30

    With the growing demand for fossil fuels and the severe energy crisis, lignocellulose is widely regarded as a promising cost-effective renewable resource for ethanol production, and the use of lignocellulose residues as raw material is remarkable. Polar organisms have important value in scientific research and development for their novelty, uniqueness and diversity. In this study, a fungus Aspergillus sydowii MS-19, with the potential for lignocellulose degradation was screened out and isolated from an Antarctic region. The growth profile of Aspergillus sydowii MS-19 was measured, revealing that Aspergillus sydowii MS-19 could utilize lignin as a sole carbon source. Its ability to synthesize low-temperature lignin peroxidase (Lip) and manganese peroxidase (Mnp) enzymes was verified, and the properties of these enzymes were also investigated. High-throughput sequencing was employed to identify and characterize the transcriptome of Aspergillus sydowii MS-19. Carbohydrate-Active Enzymes (CAZyme)-annotated genes in Aspergillus sydowii MS-19 were compared with those in the brown-rot fungus representative species, Postia placenta and Penicillium decumbens. There were 701CAZymes annotated in Aspergillus sydowii MS-19, including 17 cellulases and 19 feruloyl esterases related to lignocellulose-degradation. Remarkably, one sequence annotated as laccase was obtained, which can degrade lignin. Three peroxidase sequences sharing a similar structure with typical lignin peroxidase and manganese peroxidase were also found and annotated as haem-binding peroxidase, glutathione peroxidase and catalase-peroxidase. In this study, the fungus Aspergillus sydowii MS-19 was isolated and shown to synthesize low-temperature lignin-degrading enzymes: lignin peroxidase (Lip) and manganese peroxidase (Mnp). These findings provide useful information to improve our understanding of low-temperature lignocellulosic enzyme production by polar microorganisms and to facilitate research and

  9. Thermal Characterization of Purified Glucose Oxidase from A Newly Isolated Aspergillus Niger UAF-1

    Science.gov (United States)

    Anjum Zia, Muhammad; Khalil-ur-Rahman; K. Saeed, Muhammad; Andaleeb, Fozia; I. Rajoka, Muhammad; A. Sheikh, Munir; A. Khan, Iftikhar; I. Khan, Azeem

    2007-01-01

    An intracellular glucose oxidase was isolated from the mycelium extract of a locally isolated strain of Aspergillus niger UAF-1. The enzyme was purified to a yield of 28.43% and specific activity of 135 U mg−1 through ammonium sulfate precipitation, anion exchange and gel filtration chromatography. The enzyme showed high affinity for D-glucose with a Km value of 2.56 mM. The enzyme exhibited optimum catalytic activity at pH 5.5. Temperature optimum for glucose oxidase, catalyzed D-glucose oxidation was 40°C. The enzyme showed a high thermostability having a half-life 30 min, enthalpy of denaturation 99.66 kJ mol−1 and free energy of denaturation 103.63 kJ mol−1. These characteristics suggest the use of glucose oxidase from Aspergillus niger UAF-1 as an analytical reagent and in the design of biosensors for clinical, biochemical and diagnostic assays. PMID:18193107

  10. Antimicrobial sensitivity profile of Staphylococcus spp. Isolated from clinical mastitis

    OpenAIRE

    Thamires Martins; Adriana Frizzarin; Lívia Castelani; Heloisa Solda de Azevedo; Juliana Rodrigues Pozzi Arcaro; Cláudia Rodrigues Pozzi

    2012-01-01

    Inflammation of the mammary gland, which is also known as mastitis, occupies a prominent place among the diseases that affect dairy cattle, having a great economic importance in the dairy sector. Mastitis may have different origins, however, infectious mastitis is the most frequent and represents a risk to public health due to the propagation of microorganisms through milk. Staphylococcus spp. are considered the microorganisms that cause the greatest losses in milk production, being that Stap...

  11. Production of biosurfactant by Pseudomonas spp. isolated from industrial waste in Turkey

    OpenAIRE

    KAYA, Tayfun; ASLIM, Belma; KARİPTAŞ, Ergin

    2014-01-01

    In this study, 26 Pseudomonas spp. were isolated from a stream polluted by factory waste and from petroleum-contaminated soil. The surface tension (ST) of the cultures was used as a criterion for the primary isolation of biosurfactant-producing bacteria. Biosurfactant production was quantified by ST reduction, critical micelle concentration (CMC), emulsification capacity (EC), and cell surface hydrophobicity (CSH). Two of the isolates, P. aeruginosa 78 and 99, produced rhamnolipid biosurfacta...

  12. Antibiotic of resistence profile of Salmonella spp. serotypes isolated from retail beef in Mexico City.

    OpenAIRE

    Nova Nayarit-Ballesteros; María Salud Rubio-Lozano; Enrique Delgado-Suárez; Danilo Méndez-Medina; Diego Braña-Varela; Oscar Rodas-Suárez

    2016-01-01

    Objective. To determine the serotype and antibiotic resistance profile of Salmonella spp. isolated from retail ground beef in Mexico City. Materials and methods. A total of 100 samples of ground beef were analyzed. The pathogen was isolated by conventional methods and confirmed by PCR (invA gene, 284 bp). The antibiotic resistance profile was determined by the Kirby-Bauer method while serotyping was performed according to the Kauffman-White scheme. Results. We isolated a total of 19 strains o...

  13. Etiology and pathogenicity of two different isolates of Colletotrichum spp. obtained from physic nut seeds

    OpenAIRE

    Santos,Gil Rodrigues dos; Tozze Júnior,Hugo José; Sá,Danila Alves Corrêa de; Furtado,Gleiber Quintão; Massola Júnior,Nelson Sidnei

    2013-01-01

    The species known as physic nut (Jatropha curcas L.) has become important as one of main sources of feedstock for biodiesel production. The aims of this study were characterizing two different isolates of Colletotrichum spp. obtained from seeds of this species, through morphological, cultural, and molecular analyses; as well as assessing pathogenicity of both isolates on leaves and fruit of this plant species. For morphological analysis, length and width of 30 spores of each isolate, produced...

  14. Chemical constituents of Aspergillus sp EJC08 isolated as endophyte from Bauhinia guianensis and their antimicrobial activity.

    Science.gov (United States)

    Pinheiro, Eduardo A A; Carvalho, Josiwander M; Santos, Diellem C P dos; Feitosa, André O; Marinho, Patrícia S B; Guilhon, Giselle Maria S P; Santos, Lourivaldo S; Souza, Afonso L D de; Marinho, Andrey M R

    2013-01-01

    The present work reports the isolation of five compounds from Aspergillus sp EJC08 isolated as endophytic from Bauhinia guianensis, a tipical plant of the Amazon. The compounds ergosterol (1), ergosterol peroxide (2), mevalolactone (3), monomethylsulochrin (4) and trypacidin A (5) were isolated by chromatographic procedures and identified by spectral methods of 1D and 2D NMR and MS. Compounds 3, 4 and 5 were tested against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus and showed good activity.

  15. Optimal pcr primers for rapid and accurate detection of Aspergillus flavus isolates.

    Science.gov (United States)

    Al-Shuhaib, Mohammed Baqur S; Albakri, Ali H; Alwan, Sabah H; Almandil, Noor B; AbdulAzeez, Sayed; Borgio, J Francis

    2018-03-01

    Aspergillus flavus is among the most devastating opportunistic pathogens of several food crops including rice, due to its high production of carcinogenic aflatoxins. The presence of these organisms in economically important rice strip farming is a serious food safety concern. Several polymerase chain reaction (PCR) primers have been designed to detect this species; however, a comparative assessment of their accuracy has not been conducted. This study aims to identify the optimal diagnostic PCR primers for the identification of A. flavus, among widely available primers. We isolated 122 A. flavus native isolates from randomly collected rice strips (N = 300). We identified 109 isolates to the genus level using universal fungal PCR primer pairs. Nine pairs of primers were examined for their PCR diagnostic specificity on the 109 isolates. FLA PCR was found to be the optimal PCR primer pair for specific identification of the native isolates, over aflP(1), aflM, aflA, aflD, aflP(3), aflP(2), and aflR. The PEP primer pair was found to be the most unsuitable for A. flavus identification. In conclusion, the present study indicates the powerful specificity of the FLA PCR primer over other commonly available diagnostic primers for accurate, rapid, and large-scale identification of A. flavus native isolates. This study provides the first simple, practical comparative guide to PCR-based screening of A. flavus infection in rice strips. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Evaluation of the in vitro activity of isavuconazole and comparator voriconazole against 2635 contemporary clinical Candida and Aspergillus isolates

    DEFF Research Database (Denmark)

    Astvad, K. M.T.; Hare, R. K.; Arendrup, M. C.

    2017-01-01

    Objective The in vitro activity of isavuconazole was determined for 1677 Candida and 958 Aspergillus isolates from 2012 to 2014 with voriconazole as comparator. Methods Aspergillus isolates were screened for resistance using azole-agar. Aspergillus isolates that screened positive and all Candida...

  17. Genetic characterisation of tigecycline-resistant Enterobacter spp. in blood isolates causing bacteraemia.

    Science.gov (United States)

    Cha, Min Kyeong; Kang, Cheol-In; Park, Ga Eun; Kim, So Hyun; Chung, Doo Ryeon; Peck, Kyong Ran; Song, Jae-Hoon

    2018-01-05

    Tigecycline (TIG) is one of the most important antimicrobial agents used to treat infections by multidrug-resistant bacteria. However, rates of TIG-resistant pathogens have increased recently. This study was conducted to identify the antimicrobial susceptibility profiles and to investigate the role of efflux pumps in high-level TIG-resistant Enterobacter spp. isolates causing bacteraemia. A total of 323 Enterobacter spp. causing bacteraemia were collected from eight hospitals in various regions of South Korea. Minimum inhibitory concentrations (MICs) were determined by the broth microdilution method and Etest. Expression levels of the efflux pump gene acrA and its regulators (ramA and rarA) were examined by quantitative real-time PCR. Isolate relatedness was determined by multilocus sequence typing (MLST). Among the 323 clinical isolates included in this study, 37 (11.5%) were TIG-non-susceptible, of which 8 isolates were highly resistant to TIG with MICs of 8mg/L (4 isolates) or 16mg/L (4 isolates). All high-level TIG-resistant isolates showed increased expression of acrA (0.93-13.3-fold) and ramA (1.4-8.2-fold). Isolates with a tigecycline MIC of 16mg/L also showed overexpression of rarA compared with TIG-susceptible isolates. In this study, overexpression of acrA, ramA and rarA was observed in high-level TIG-resistant Enterobacter spp. isolates. We suggest that rarA might be involved in the regulation of acrA overexpression in high-level TIG-resistant Enterobacter spp. isolates. Efflux pump-mediated resistance should be closely monitored because it could be indirectly attributed to the use of other antibiotics transported by the same efflux pump. Copyright © 2017. Published by Elsevier Ltd.

  18. Presentación sistémica de Aspergillus spp con semiología neurológica en un Pastor Alemán: informe de un caso clínico

    OpenAIRE

    Hernández Tapia, Ana Laura; Santoscoy Mejía, Eduardo Carlos; Ramírez Lezama, José; Cervantes Olivares, Roberto A.; Quiroz Mercado, Joaquín Arturo

    2010-01-01

    La infección del sistema nervioso en el perro es poco común; posibles causas son Aspergillus spp, Cryptococcus neoformans, Blastomyces dermatidis y Coccidioides immitis. La infección por Aspergillus spp afecta al aparato respiratorio y en pocas ocasiones se presenta de manera diseminada. Sin embargo, en el Pastor Alemán hay mayor incidencia de éste, pues dicha infección se asocia con una deficiencia inmunológica. Se describe el caso de una perra de raza Pastor Alemán, de 1.5 años de edad, ref...

  19. Plastic degrading fungi Trichoderma viride and Aspergillus nomius isolated from local landfill soil in Medan

    Science.gov (United States)

    Munir, E.; Harefa, R. S. M.; Priyani, N.; Suryanto, D.

    2018-03-01

    Plastic is a naturally recalcitrant polymer, once it enters the environment, it will remain there for many years. Accumulation of plastic as wastes in the environment poses a serious problem and causes an ecological threat. Alternative strategies to reduce accumulation of plastic wastes have been initiated and implemented from a different aspect including from microbiological view point. The study to obtain potential fungi in degrading plastic molecule has been initiated in our laboratory. Low density polyethylene (LDPE) plastic was used as a tested material. Candidate fungi were isolated from local landfill soil. The fungi were cultured in mineral salt medium broth containing LDPE powder. Two of nine isolates showed best growth response in broth media containing LDPE. These isolates (RH03 and RH06) were used in degradation test. Results showed that isolate RH03 and RH06 reduced the weight of LDPE film by 5.13% and 6.63%, respectively after 45 days of cultivation. The tensile strength of treated film even reduced significantly by 58% and 40% of each isolate. Analyses of electron micrograph exhibited grove ands rough were formed on the surface of LDPE film. These were not found in the untreated film. Furthermore, molecular analysis through polymerase chain reaction and DNA sequencing indicated that RH03 is Trichoderma viride and RH06 is Aspergillus nomius with 97% and 96% similarities, respectively.

  20. Characterisation of Bergeyella spp. isolated from the nasal cavities of piglets.

    Science.gov (United States)

    Lorenzo de Arriba, M; Lopez-Serrano, S; Galofre-Mila, N; Aragon, V

    2018-04-01

    The aim of this study was to characterise bacteria in the genus Bergeyella isolated from the nasal passages of healthy piglets. Nasal swabs from 3 to 4 week-old piglets from eight commercial domestic pig farms and one wild boar farm were cultured under aerobic conditions. Twenty-nine Bergeyella spp. isolates were identified by partial 16S rRNA gene sequencing and 11 genotypes were discriminated by enterobacterial repetitive intergenic consensus (ERIC)-PCR. Bergeyella zoohelcum and Bergeyella porcorum were identified within the 11 genotypes. Bergeyella spp. isolates exhibited resistance to serum complement and phagocytosis, poor capacity to form biofilms and were able to adhere to epithelial cells. Maneval staining was consistent with the presence of a capsule. Multiple drug resistance (resistance to three or more classes of antimicrobial agents) was present in 9/11 genotypes, including one genotype isolated from wild boar with no history of antimicrobial use. In conclusion, Bergeyella spp. isolates from the nasal cavities of piglets showed some in vitro features indicative of a potential for virulence. Further studies are necessary to identify the role of Bergeyella spp. in disease and within the nasal microbiota of pigs. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  1. Isolation, Identification and Antibacterial Susceptibility of Staphylococcus spp. Associated with the Mobile Phones of University Students.

    Science.gov (United States)

    Furuhata, Katsunori; Ishizaki, Naoto; Sogawa, Kazuyuki; Kawakami, Yasushi; Lee, Shin-Ichi; Sato, Masahiro; Fukuyama, Masafumi

    2016-01-01

    From May 2014 to February 2015, 319 university students (male, n=173; female n=146) of 18 to 24 years of age who carried mobile phones or computer tablets were selected as subjects. Staphylococcus spp. were detected in 101 of 319 samples (31.7%). In the present study, 11 strains of S. aureus were isolated and identified, not all of which were methicillin-resistant Staphylococcus aureus (MRSA). Overall, 14 species were identified, with 11 strains (10.9%) of S. xylosus being isolated at the highest frequency. Following this were eight strains (7.9%) of S. cohnii and seven strains (6.9%) each of S. capitis and S. haemolyticus. Staphylococcus spp. isolation was performed with bacterial samples obtained from the mobile phones of 22 specific subjects (males, n=12; females, n=10). Staphylococcus spp. isolation was performed on days -1, 7 and 30 of the experiment. Staphylococcus spp. were positively detected one or more times in 12 subjects (54.5%). In one subject (8.3%), all three tests were positive. Furthermore, two tests were positive in three (25.0%). In the eight remaining subjects (66.7%) Staphylococcus spp. were detected only once. For the three abovementioned tests, we investigated the pulsed-field gel electrophoresis (PFGE) patterns of the strains derived from the mobile phone and from the fingers of three subjects in whom the same bacterial species were isolated twice. From the cases with similarities between strains derived from the fingers and the mobile phones and cases, with consistency in the strains derived from the mobile phone at different times, commonality was observed in the strains derived from the fingers and mobile phones along with chronological uniformity in the strains derived from the mobile phones. A total of 101 Staphylococcus spp. strains were isolated from mobile phones. According to drug susceptibility tests, 99 strains (98.0%) were found to have some degree of resistance to drugs (excluding one strain each of S. aureus and S. haemolyticus

  2. Isolation and NMR Characterization of Fumonisin B-2 and a New Fumonisin B-6 from Aspergillus niger

    DEFF Research Database (Denmark)

    Månsson, Maria; Klejnstrup, Marie Louise; Phipps, Richard Kerry

    2010-01-01

    A new fumonisin, fumonisin B-6 (1), has been isolated by cation-exchange and reverse-phase chromatography, together with fumonisin B-2 (2), from,stationary cultures of the fungus Aspergillus niger NRRL 326. Analysis of mass spectrometric and NMR data determined that FB6 is a positional isomer...

  3. Cloning and Genomic Organization of a Rhamnogalacturonase Gene from Locally Isolated Strain of Aspergillus niger.

    Science.gov (United States)

    Damak, Naourez; Abdeljalil, Salma; Taeib, Noomen Hadj; Gargouri, Ali

    2015-08-01

    The rhg gene encoding a rhamnogalacturonase was isolated from the novel strain A1 of Aspergillus niger. It consists of an ORF of 1.505 kb encoding a putative protein of 446 amino acids with a predicted molecular mass of 47 kDa, belonging to the family 28 of glycosyl hydrolases. The nature and position of amino acids comprising the active site as well as the three-dimensional structure were well conserved between the A. niger CTM10548 and fungal rhamnogalacturonases. The coding region of the rhg gene is interrupted by three short introns of 56 (introns 1 and 3) and 52 (intron 2) bp in length. The comparison of the peptide sequence with A. niger rhg sequences revealed that the A1 rhg should be an endo-rhamnogalacturonases, more homologous to rhg A than rhg B A. niger known enzymes. The comparison of rhg nucleotide sequence from A. niger A1 with rhg A from A. niger shows several base changes. Most of these changes (59 %) are located at the third base of codons suggesting maintaining the same enzyme function. We used the rhamnogalacturonase A from Aspergillus aculeatus as a template to build a structural model of rhg A1 that adopted a right-handed parallel β-helix.

  4. Effects of Hydrogen Peroxide on Different Toxigenic and Atoxigenic Isolates of Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Jake C. Fountain

    2015-08-01

    Full Text Available Drought stress in the field has been shown to exacerbate aflatoxin contamination of maize and peanut. Drought and heat stress also produce reactive oxygen species (ROS in plant tissues. Given the potential correlation between ROS and exacerbated aflatoxin production under drought and heat stress, the objectives of this study were to examine the effects of hydrogen peroxide (H2O2-induced oxidative stress on the growth of different toxigenic (+ and atoxigenic (− isolates of Aspergillus flavus and to test whether aflatoxin production affects the H2O2 concentrations that the isolates could survive. Ten isolates were tested: NRRL3357 (+, A9 (+, AF13 (+, Tox4 (+, A1 (−, K49 (−, K54A (−, AF36 (−, and Aflaguard (−; and one A. parasiticus isolate, NRRL2999 (+. These isolates were cultured under a H2O2 gradient ranging from 0 to 50 mM in two different media, aflatoxin-conducive yeast extract-sucrose (YES and non-conducive yeast extract-peptone (YEP. Fungal growth was inhibited at a high H2O2 concentration, but specific isolates grew well at different H2O2 concentrations. Generally the toxigenic isolates tolerated higher concentrations than did atoxigenic isolates. Increasing H2O2 concentrations in the media resulted in elevated aflatoxin production in toxigenic isolates. In YEP media, the higher concentration of peptone (15% partially inactivated the H2O2 in the media. In the 1% peptone media, YEP did not affect the H2O2 concentrations that the isolates could survive in comparison with YES media, without aflatoxin production. It is interesting to note that the commercial biocontrol isolates, AF36 (−, and Aflaguard (−, survived at higher levels of stress than other atoxigenic isolates, suggesting that this testing method could potentially be of use in the selection of biocontrol isolates. Further studies will be needed to investigate the mechanisms behind the variability among isolates with regard to their degree of oxidative stress

  5. Comprehensive update of dalbavancin activity when tested against uncommonly isolated streptococci, Corynebacterium spp., Listeria monocytogenes, and Micrococcus spp. (1357 strains).

    Science.gov (United States)

    Jones, Ronald N; Stilwell, Matthew G

    2013-06-01

    Dalbavancin is an investigational lipoglycopeptide having an extended serum elimination half-life allowing once-weekly dosing. Data from testing 1357 strains of uncommonly isolated species expand the dalbavancin spectrum details as follows (MIC50/90): β-haemolytic streptococcal serogroups C, F, and G (≤0.03/≤0.03 μg/mL), 7 viridans group of streptococci (≤0.03/≤0.03-0.06 μg/mL), 5 Corynebacterium spp. (0.06/0.12 μg/mL), Listeria monocytogenes (0.06/0.12 μg/mL), and Micrococcus spp. (≤0.03/≤0.03 μg/mL). Among all reported isolates, 99.8% of tested strains were inhibited at dalbavancin MIC values at ≤0.12 μg/mL. Dalbavancin remains very potent against rarer Gram-positive pathogens, using in vitro test experience with organisms cultured through 2011. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Genetic characterization of trh positive Vibrio spp. isolated from Norway

    Directory of Open Access Journals (Sweden)

    Anette eBauer Ellingsen

    2013-12-01

    Full Text Available The thermostable direct hemolysin (TDH and/or TDH-related hemolysin (TRH genes are carried by most virulent Vibrio parahaemolyticus serovars. In Norway, trh+ V. parahaemolyticus constitute 4.4% and 4.5 % of the total number of V. parahaemolyticus isolated from blue mussel (Mytilus edulis and water, respectively. The trh gene is located in a region close to the gene cluster for urease production (ure. This region was characterized in V. parahaemolyticus strain TH3996 and it was found that a nickel transport operon (nik was located between the first gene (ureR and the rest of the ure cluster genes. The organization of the trh-ureR-nik-ure gene cluster in the Norwegian trh+ isolates was unknown. In this study, we explore the gene organization within the trh-ureR-nik-ure cluster for these isolates. PCR analyses revealed that the genes within the trh-ureR-nik-ure gene cluster of Norwegian trh+ isolates were organized in a similar fashion as reported previously for TH33996. Additionally, the phylogenetic relationship among these trh+ isolates was investigated using Multilocus Sequence Typing (MLST. Analysis by MLST or ureR-trh sequences generated two different phylogenetic trees for the same strains analyzed, suggesting that ureR-trh genes have been acquired at different times in Norwegian V. parahaemolyticus isolates. MLST results revealed that some pathogenic and non-pathogenic V. parahaemolyticus isolates in Norway appear to be highly genetically related.

  7. Bioremediation of coractive blue dye by using Pseudomonas spp. isolated from the textile dye wastewater

    Science.gov (United States)

    Sunar, N. M.; Mon, Z. K.; Rahim, N. A.; Leman, A. M.; Airish, N. A. M.; Khalid, A.; Ali, R.; Zaidi, E.; Azhar, A. T. S.

    2018-04-01

    Wastewater released from the textile industry contains variety substances, mainly dyes that contains a high concentration of color and organic. In this study the potential for bacterial decolorization of coractive blue dye was examined that isolated from textile wastewater. The optimum conditions were determined for pH, temperature and initial concentration of the dye. The bacteria isolated was Pseudomonas spp. The selected bacterium shows high decolorization in static condition at an optimum of pH 7.0. The Pseudomonas spp. could decolorize coractive blue dye by 70% within 24 h under static condition, with the optimum of pH 7.0. Decolorization was confirmed by using UV-VIS spectrophotometer. This present study suggests the potential of Pseudomonas spp. as an approach in sustainable bioremediation that provide an efficient method for decolorizing coractive blue dye.

  8. Phospholipase and proteinase activities of Candida spp. isolates from vulvovaginitis in Iran.

    Science.gov (United States)

    Shirkhani, S; Sepahvand, A; Mirzaee, M; Anbari, K

    2016-09-01

    This study aims to characterize phospholipase and proteinase activities of Candida isolates from 82 vulvovaginal candidiasis (VVC) and to study the relationship of these activities with vulvovaginitis. Totally 82 Candida isolates from vagina samples of VVC patients were randomly collected over the period between September and December 2014 from hospitalized patients at the general hospitals of Lorestan province, Iran. Isolates were previously identified by conventional mycological methods. The phospholipase and proteinase activities were evaluated by Egg yolk agar, Tween 80 opacity medium and agar plate methods. The most common Candida species was identified Candida albicans (n=34, 41.5%), followed by Candida famata (n=13, 15.8%), Candida tropicalis (n=11, 13.4%), and Candida parapsilosis (n=9, 11%). The most phospholipase activity was observed in Candida colliculosa (40%), followed by C. famata (38.5%), and Candida krusei (33.3%). The findings revealed that the correlation between phospholipase production by Candida spp. and the presence of VVC was not found to be statistically significant (P=0.91). All Candida spp. exhibited considerable proteinase activity; so that 100% of C. colliculosa, C. parapsilosis, Candida kefyr, and Candida intermedia isolates produced high proteinase activity with Pz 4+ scores. There was a significant correlation between proteinase production by Candida spp. and the presence of VVC (P=0.009). The obtained findings revealed that Candida spp. isolates may produce both virulence factors, phospholipase and proteinase. Although the phospholipase production was only observed in <40% of the isolates; however there was a significant association between proteinase production by Candida spp. and VVC. Copyright © 2016. Published by Elsevier Masson SAS.

  9. Aflatoxins produced by Aspergillus nomius ASR3, a pathogen isolated from the leaf-cutter ant Atta sexdens rubropilosa

    Directory of Open Access Journals (Sweden)

    Eduardo Afonso da Silva-Junior

    Full Text Available ABSTRACT Aspergillus spp. cause economic impacts due to aflatoxins production. Although the toxicity of aflatoxins is already known, little information about their ecological roles is available. Here we investigated the compounds produced by Aspergillus nomius ASR3 directly from a dead leaf-cutter queen ant Atta sexdens rubropilosa and the fungal axenic culture. Chemical analyses were carried out by high-resolution mass spectrometry and tandem mass spectrometry techniques. Aflatoxins B1 and G1 were detected in both the axenic culture and in the dead leaf-cutter queen ant. The presence of these mycotoxins in the dead leaf-cutter queen ant suggests that these compounds can be related to the insect pathogenicity of A. nomius against A. sexdens rubropilosa.

  10. Antibiotic of resistence profile of Salmonella spp. serotypes isolated from retail beef in Mexico City.

    Directory of Open Access Journals (Sweden)

    Nova Nayarit-Ballesteros

    2016-05-01

    Full Text Available Objective. To determine the serotype and antibiotic resistance profile of Salmonella spp. isolated from retail ground beef in Mexico City. Materials and methods. A total of 100 samples of ground beef were analyzed. The pathogen was isolated by conventional methods and confirmed by PCR (invA gene, 284 bp. The antibiotic resistance profile was determined by the Kirby-Bauer method while serotyping was performed according to the Kauffman-White scheme. Results. We isolated a total of 19 strains of Lomita (6, Derby (4, Senftenberg (2, Javiana and Cannsttat (1 and undeter- mined (5 serotypes. The strains showed a high resistance rate to ampicillin (18/19, carbenicillin (16/19, tetracyclin (13/19, and trimethoprim-sulfamethoxazole (13/19. Multidrug resistance was observed in 14 isolates. Conclusions. Several Salmonella spp. serotypes of public health significance are circulating in ground beef sold in the major Mexican city. Some of these strains are multi-drug resistance.

  11. Isolation and characterization of Campylobacter spp. from Antarctic fur seals (Arctocephalus gazella) at Deception Island, Antarctica.

    Science.gov (United States)

    García-Peña, F J; Pérez-Boto, D; Jiménez, C; San Miguel, E; Echeita, A; Rengifo-Herrera, C; García-Párraga, D; Ortega-Mora, L M; Pedraza-Díaz, S

    2010-09-01

    The presence of Campylobacter spp. was investigated in 41 Antarctic fur seals (Arctocephalus gazella) and 9 Weddell seals (Leptonychotes weddellii) at Deception Island, Antarctica. Infections were encountered in six Antarctic fur seals. The isolates, the first reported from marine mammals in the Antarctic region, were identified as Campylobacter insulaenigrae and Campylobacter lari.

  12. Effect of Plant Essential Oils and Gamma Irradiation on Growth and Aflatoxin Production by Aspergillus Flavus Isolated from Wheat Grains

    International Nuclear Information System (INIS)

    Salem, E.A.; Shalaby, Kh.

    2016-01-01

    The antifungal potential of essential oils of Thyme (Thymus vulgaris L.) and camphor ( Eucalyptus rostrata L.) was determined on Aspergillus flavus link isolated from wheat grains on Potato dextrose agar (PDA). They inhibited completely mycelia growth of the fungus at 1000 and 2000 ppm, and prevented aflatoxin production at sub lethal dose 500 and 1000 ppm respectively. Gamma radiation was used to control mycelia growth of Aspergillus flavus Link and inhibiting aflatoxin production. A dose level of 3.5 KGy gamma radiation prevented the fungal growth and aflatoxin production by A. flavus link, where a dose of 2.5 K Gy ( the sub lethal dose) prevented about 85% of aflatoxin production

  13. Morphological Characterization and Determination of Aflatoxin-Production Potentials of Aspergillus flavus Isolated from Maize and Soil in Kenya

    OpenAIRE

    Matome Gabriel Thathana; Hunja Murage; Akebe Luther King Abia; Michael Pillay

    2017-01-01

    This study aimed at morphologically identifying Aspergillus flavus in soil and maize and at determining their aflatoxin-producing potentials. Five hundred and fourteen isolates obtained from maize and soil in Kenya were cultivated on Czapeck Dox Agar, Malt Extract Agar, Sabouraud Dextrose Agar, Potato Dextrose Agar, and Rose-Bengal Chloramphenicol Agar. Isolates were identified using macro-morphological characteristics. Micromorphological characteristics were determined using slide cultures. ...

  14. Isolation and genetic analysis of Aspergillus niger mutants with reduced extracellular glucoamylase

    International Nuclear Information System (INIS)

    Valent, G.U.; Calil, M.R.; Bonatelli Junior, R.

    1992-01-01

    Mutants with impaired production of extracellular glucoamylase were isolated at a high frequency (2% of survivors) from an Aspergillus niger strain treated with UV light. These were designated as low glucoamylase producers (lgp, up to 30% of the parental yield) and medium producers (mgp, a 35 to 50% decrease in enzyme level). All the mutants were shown to be recessive; one strain segregated two unlinked genes. Complementation tests, and segregation from heterozygous diploid, suggested at least three to four unlinked genes, each able to impair glucoamylase production. There is evidence of a single structural gene for glucoamylase in A. niger. Therefore, as production of extracellular enzymes is normally the final result of several steps at intracellular and membrane levels, including regulation of enzyme synthesis, we suggest intergenic interaction that controls extracellular enzyme accumulation and that mutation in any of these genes would result in impaired production. (author)

  15. Cyclo-(trp-phe diketopiperazines from the endophytic fungus Aspergillus versicolor isolated from Piper aduncum

    Directory of Open Access Journals (Sweden)

    Juliana R. Gubiani

    Full Text Available Six known compounds, three peptide derivatives: cyclo-(tryptophyl-phenylalanyl (2, diketopiperazine dimer WIN 64821 (3 and 3-hydroxy-15H-tryptophenaline (4, one adenine derivative: 2-hydroxy-6-N-isopentenyl-adenine (5, one phtalide derivative: 4-methoxyphtalide (1 and one benzoic acid derivative: 3-hydroxy-4-(1-hydroxy-1,5-dimethyl-hexyl benzoic acid (6, were isolated from the ethyl acetate extract of the endophytic fungus Aspergillus versicolor associated with the Piper aduncum plant. Their structures were determined on the basis of detailed interpretation of 1 D and 2D NMR spectra and in comparison with works reported in the literature. This paper, in effect, deals with the first report of these compounds in A. versicolor.

  16. Biopathologic Characterization of Three Mixed Poultry Eimeria spp. Isolates

    Directory of Open Access Journals (Sweden)

    B Mansoori

    2011-09-01

    Full Text Available Background: Coccidiosis of domestic fowl, caused by species of the Genus Eimeria, is responsi­ble for important economic losses in poultry production. Because different species and/or strains can vary in pathogenicity and other biological parameters, their precise characteriza­tion is important for epizootiological studies.Methods: Fifty samples from litter, whole intestinal tract and feces were collected from poul­try houses located in different provinces of Iran. One hundred twenty male day-old broi­ler chicks were challenged with three selected isolates. Data on weight gain, Food Conversion Ratio (FCR, food intake, lesion scoring and shedding of oocysts per gram of feces were rec­orded and analyzed by the Duncan's test.Results: In all treatments, the challenged groups had statistically significant lower weight gain than that of unchallenged control group. Isolate three caused the lowest weight gain and food intake and the worst lesion score as well as FCR. Despite originating from close geographi­cal regions for isolates 1 and 2, the difference in biopathologic factors may be either due to different proportion of identified species or the different pathogenicity of the species present in the isolates.Conclusion: The results highlight the importance of considering various species of Eimeria in designing the preventive, control and treatment strategies to prevent coccidiosis in different regions of Iran. Further characterization of each isolate would be the next step to provide a basis for coccidiosis research with well-characterized local isolates.

  17. Aspergillus otitis in small animals--a retrospective study of 17 cases.

    Science.gov (United States)

    Goodale, Elizabeth C; Outerbridge, Catherine A; White, Stephen D

    2016-02-01

    Aspergillus spp. are saprophytic opportunistic fungal organisms and are a common cause of otomycosis in humans. Although there have been case reports of Aspergillus otitis externa in dogs, to the best of the authors' knowledge, this is the first retrospective case series describing Aspergillus otitis in dogs and cats. To characterize signalment, putative risk factors, treatments and outcomes of a case series of dogs and cats with Aspergillus otitis. Eight dogs and nine cats diagnosed with Aspergillus otitis. A retrospective review of medical records from 1989 to 2014 identified animals diagnosed with Aspergillus otitis based on culture. All dogs weighed greater than 23 kg. The most common putative risk factors identified in this study were concurrent diseases, therapy causing immunosuppression or a history of an otic foreign body. Aspergillus otitis was unilateral in all study dogs and most cats. Concurrent otitis media was confirmed in three dogs and one cat, and suspected in two additional cats. Aspergillus fumigatus was the most common isolate overall and was the dominant isolate in cats. Aspergillus niger and A. terreus were more commonly isolated from dogs. Animals received various topical and systemic antifungal medications; however, otic lavage under anaesthesia and/or surgical intervention increased the likelihood of resolution of the fungal infection. Aspergillus otitis is uncommon, typically seen as unilateral otitis externa in cats and larger breed dogs with possible risk factors that include immunosuppression and otic foreign bodies; previous antibiotic usage was common. © 2015 ESVD and ACVD.

  18. Natamycin and Azithromycin are Synergistic in vitro against Ocular Pathogenic Aspergillus flavus species complex and Fusarium solani species complex Isolates.

    Science.gov (United States)

    Guo, Haoyi; Zhou, Lutan; He, Yi; Gao, Chuanwen; Han, Lei; Xu, Yan

    2018-05-07

    The interaction of natamycin-azithromycin combination against 60 ocular fungal isolates was tested in vitro The combination produced 100% synergistic interactions when natamycin added azithromycin at 20, 40, 50 μg/ml against Aspergillus flavus species complex (AFSC) isolates and added azithromycin at 50 μg/ml against Fusarium solani species complex isolates. The combination with 50 μg/ml azithromycin enhanced natamycin's effect against AFSC isolates by reducing natamycin MICs from MIC 90 64μg/ml to MIC 90 0.031μg/ml. No antagonism was observed. Copyright © 2018 American Society for Microbiology.

  19. Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals

    Energy Technology Data Exchange (ETDEWEB)

    Buckova, M.; Godocikova, J.; Simonovicova, A.; Polek, B. [Slovakian Academy of Science, Bratislava (Slovakia)

    2005-04-15

    Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As{sup 5+}, Cd{sup 2+}, Cu{sup 2+}) at final concentrations of 25 and 50 mg/L and H{sub 2}O{sub 2} (20 or 40 m(M)) mostly stimulated production of catalases only in isolates from mines surroundings, and H{sub 2}O{sub 2} and Hg{sup 2+} caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H{sub 2}O{sub 2}, as monitored by growth, than did the strain from the culture collection.

  20. Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.

    Directory of Open Access Journals (Sweden)

    Dong Ying Wang

    Full Text Available Aspergillus flavus is second only to A. fumigatus in causing invasive aspergillosis and it is the major agent responsible for fungal sinusitis, keratitis and endophthalmitis in many countries in the Middle East, Africa and Southeast Asia. Despite the growing challenge due to A. flavus, data on the molecular epidemiology of this fungus remain scarce. The objective of the present study was to develop a new typing method based on the detection of VNTR (Variable number tandem repeat markers. Eight VNTR markers located on 6 different chromosomes (1, 2, 3, 5, 7 and 8 of A. flavus were selected, combined by pairs for multiplex amplifications and tested on 30 unrelated isolates and six reference strains. The Simpson index for individual markers ranged from 0.398 to 0.818. A combined loci index calculated with all the markers yielded an index of 0.998. The MLVA (Multiple Locus VNTR Analysis technique proved to be specific and reproducible. In a second time, a total of 55 isolates from Chinese avian farms and from a Tunisian hospital have been evaluated. One major cluster of genotypes could be defined by using the graphing algorithm termed Minimum Spanning Tree. This cluster comprised most of the isolates collected in an avian farm in southern China. The MLVA technique should be considered as an excellent and cost-effective typing method that could be used in many laboratories without the need for sophisticated equipment.

  1. Optimization of pectinase enzyme production in Aspergillus fumigatus isolated from rotten fruits

    Directory of Open Access Journals (Sweden)

    2015-12-01

    Full Text Available Introduction: Pectinase is one of the most important industrial enzymes which was isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the juice and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi have been done by plate culture on pectin medium and staining with Lugol's iodine solution. The best strain was identified by method of Pitt and Hocking as Aspergillus fumigates. The enzyme production was optimized by application of the factorial design which involves five factors, each at three levels. Five factors were carbon sources (whey, sugar, stevia and ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results showed that the optimum condition for enzyme production was at 32 °C, PH = 6 , 3g / L manganese sulfate, 2.75g / L of ammonium sulfate, 10g / L of each carbon source (whey, stevia, and glucose. Optimum of enzyme production was observed in the presence of 1.328 mg / ml of glucose. Molecular weight of enzyme was obtained about 40 kDa by SDS-PAGE. Discussion and conclusion: The results demonstrated that this strain could grow in a wide range of carbon sources, PH and temperature. This study indicates that this strain is a good candidate for use in industrial application.

  2. Data set for the mass spectrometry based exoproteome analysis of Aspergillus flavus isolates

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    Ramu Muthu Selvam

    2015-03-01

    Full Text Available Aspergillus flavus is one of the predominant causative organisms of mycotic keratitis in tropical parts of the world. Extracellular proteins are the earliest proteins that come in contact with the host and have a role in the infection process. Exoproteins of A. flavus isolated from infected cornea, sputum and a saprophyte were pooled and identified using high resolution mass spectrometry in order to get the total exoproteome from cultures isolated from different sources. A total of 637 proteins was identified from the pooled A. flavus exoproteome. Analysis based on GO annotations of the 637 identified proteins revealed that hydrolases form the predominant class of proteins in the exoproteome. Interestingly, a greater proportion of the exoproteins seem to be secreted through the non-classical pathways. This data represent the first in-depth analysis of the representative A. flavus exoproteome of a large set of isolates from distinct sources. This data have been deposited to the ProteomeXchange with identifier PXD001296.

  3. Antimicrobial resistance in E. coli and Salmonella spp. isolates from calves in southern Chile

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    Luis Hervé-Claude

    2017-09-01

    Full Text Available Objective: Description of antimicrobial resistance in E. coli and Salmonella spp. isolates from calves <30 days of age from southern Chile. Material and methods: Necropsy and microbiology reports of 107 calves <30 days of age received at the Animal Pathology Institute between 2002 and 2015 were considered. Additionally, an antimicrobial resistance score was generated to allow comparisons among isolates with different antimicrobial susceptibility profiles. Results: There was no clear trend in antimicrobial resistance during the study period, with similar levels of resistance for E. coli, β-hemolytic E. coli and Salmonella spp. Approximately 50% of isolates were sensitive to antimicrobials, and between 19 and 36% of samples showed possible extended- or pan- drug resistance. Multiple different antimicrobial resistance patterns were found, including 32 for E. coli, 17 for β-hemolytic E. coli and 10 for Salmonella spp. Conclusions: Overall, E. coli samples were most sensitive to ceftriaxone; β-hemolytic E. coli to florfenicol; and Salmonella spp. to gentamicin. In contrast, these agents were resistant to amoxicillin, ampicillin and oxytetracycline respectively. This study is unique in its approach and provides useful information for veterinarians and producers on the antibiotic resistance patterns of bacteria posing a serious threat to calves. These results can help field veterinarians to control and treat bacterial diarrhea in calves.

  4. Biodiversity and ochratoxin A profile of Aspergillus section Nigri populations isolated from wine grapes in Cyprus vineyards.

    Science.gov (United States)

    Pantelides, Iakovos S; Aristeidou, Efi; Lazari, Maria; Tsolakidou, Maria-Dimitra; Tsaltas, Dimitris; Christofidou, Maria; Kafouris, Demetris; Christou, Eftychia; Ioannou, Nicolas

    2017-10-01

    The objective of this study was to evaluate the biodiversity of Aspergillus section Nigri populations from Cyprus vineyards by morphological, toxigenic and phylogenetic analysis. Aspergillus section Nigri populations were isolated from grapes of the varieties 'Maratheftiko' and 'Cabernet Sauvignon' originating from six growing regions of Cyprus during 2010 and 2011 years. The isolation frequency of Aspergillus section Nigri from grape samples was 43.3% and a total of 284 isolates were selected for further analyses based on the macroscopic characteristics of black aspergilli. The isolates were characterized by sequencing analysis of the calmodulin gene in order to identify species responsible for ochratoxin A (OTA) production. The phylogenetic analysis showed that the isolates were grouped in three major clusters. The A. tubingensis cluster included 262 isolates (92.25%), the A. niger cluster included 15 isolates identified as A. niger (5.3%) and 6 isolates identified as A. welwitschiae (2.1%). One isolate was classified as A. carbonarius (0.35%) and was grouped in a cluster together with the reference isolates of A. carbonarius, A. sclerotioniger, A. sclerotiocarbonarius and A. ibericus. All the isolates were evaluated for their ochratoxigenic ability by HPLC coupled with a fluorescence detector (HPLC-FLD) and the positive isolates were re-examined using ultra high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). The Aspergillus carbonarius isolate produced an average quantity of 1436.1 ng OTA/g Czapek Yeast Agar (CYA); From the A. niger strains three isolates (20%) produced OTA and only one isolate from A. welwitschiae (16.7%) was proved ochratoxigenic with toxin production average at 23.9 ng/g and 9.1 ng/g CYA respectively. Grape must samples derived from the collected berries were also analyzed for OTA and none of the samples were found contaminated with the mycotoxin. The results showed that the geographic area and the

  5. First Report of an Atypical New Aspergillus parasiticus Isolates with Nucleotides Insertion in aflR Gene Identical to Aspergillus sojae

    Science.gov (United States)

    Aflatoxins are toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus favus and Aspergillus parasitic and cause toxin contamination in food chain worldwide. Aspergillus oryzae and Aspergillus sojae are highly valued as koji molds in the traditional prep...

  6. Antibiotic resistance among Ureaplasma spp. isolates: cause for concern?

    Science.gov (United States)

    Beeton, M L; Spiller, O B

    2017-02-01

    There is growing global concern regarding the rise of antibiotic-resistant organisms. Many of these reports have focused on various Gram-positive and Gram-negative pathogens, with little attention to the genus Ureaplasma. Ureaplasma spp. are associated with numerous infectious diseases affecting pregnant women, neonates and the immunocompromised. Treatment options are extremely limited due to high levels of intrinsic resistance resulting from the unique physiology of these organisms and further restricted in cases of the developing fetus or neonate, often limiting therapeutic options to predominantly macrolides or rarely fluoroquinolones. The increasing presence of macrolide- and fluoroquinolone-resistant strains among neonatal infections may result in pan-drug resistance and potentially untreatable conditions. Here, we review the requirements for accurate measurement of antimicrobial susceptibility, provide a comprehensive review of the antimicrobial resistance (AMR) for Ureaplasma species in the literature and contextualize these results relative to some investigators' reliance on commercial kits that are not CLSI compliant when determining AMR. The dramatic variation in the resistance patterns and impact of high levels of AMR amongst neonatal populations suggests the need for continued surveillance. Commercial kits represent an excellent tool for initial antibiotic susceptibility determination and screening. However, AMR reporting must utilize internationally standardized methods, as high-titre samples, or Mycoplasma hominis-contaminated samples routinely give false AMR results. Furthermore, there is a requirement for future reports to determine the underlying AMR mechanisms and determine whether expanding AMR is due to spontaneous mutation, transmission of resistance genes on mobile elements or selection and expansion of resistant clones. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy

  7. Characterization of probiotic Lactobacillus spp. isolates from commercial fermented milks

    OpenAIRE

    Farahmand, Nasim

    2015-01-01

    The aim of this project was to study the identity of probiotic lactobacilli in fermented milk products from the United Kingdom/European markets during their survival during shelf-life. This in vitro study was also aimed at undertaking studies on some of the physiological probiotic criteria, such as resistance to stomach/intestine conditions and also possible functional properties of the isolates, such as antimicrobial activities, antibiotic resistance/susceptibility and antibiotic resistance ...

  8. Biodegradation of wheat straw by different isolates of Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    A.R. Astaraei

    2016-04-01

    Full Text Available Efficient use of agricultural wastes due to their recycling and possible production of cost effective materials, have economic and ecological advantages. A biological method used for degrading agricultural wastes is a new method for improving the digestibility of these materials and favoring the ease of degradation by other microorganisms. This research was carried out to study the possible biodegradation of wheat straw by different species and isolates of Trichoderma fungi. Two weeks after inoculation of wheat straw by different isolates, oven drying in 75◦C, the samples were weighted and (Acid Detergent Fiber ADF and NDF (Neutral Detergent Fiber reductions of each sample under influence of fungal growth were compared with their controls. The results showed that biodegradation of wheat straw were closely related to fungi species and also its isolates. The Reductions in NDF and ADF of wheat straw by T. reesei and T. longibrachiatum were more pronounced compared to others, although T. reesei was superior in ADF of wheat straw reduction. It is concluded that for improving in digestibility and also shortening the timing of composting process, it is recommended to treat the wheat straw with Trichoderma fungi and especially with T. reesei and T. longibrachiatum that performed well and had excellent efficiencies.

  9. Phenotypic and molecular characterization of Klebsiella spp. isolates causing community-acquired infections

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    U. Garza-Ramos

    2018-05-01

    Full Text Available Klebsiella spp. isolates from community-acquired infections were characterized. A total of 39 Klebsiella spp. isolates were obtained from outpatients at four rural hospitals in Mexico (2013–2014. The biochemical tests identified all as being K. pneumoniae. The molecular multiplex-PCR test identified 36 (92.4% K. pneumoniae isolates and one (2.5% K. variicola isolate, and phylogenetic analysis of the rpoB gene identified two isolates (5.1% belonging to K. quasipneumoniae subsp. quasipneumoniae and K. quasivariicola. The last one was confirmed by phylogenetic analysis of six-loci concatenated genes. Mostly the isolates were multidrug resistant; however, a minority were extended-spectrum β-lactamase producing (10.2%. The extended-spectrum β-lactamase CTX-M-15 gene was identified in these isolates. Analysis of biofilm production and the hypermucoviscosity phenotype showed a total of 35 (92.3% and seven (17.9% of the isolates were positive for these phenotypes respectively. The K2 (4/39, 10.2%, K5 (2/39, 5.1% and K54 (1/39, 2.5% serotypes were identified in seven (17.9% of the isolates, and only 28.5% (2/7 hypermucoviscous isolates were positive for the K2 and K5 serotypes. In general, the sequence type (ST analysis and phylogenetic analysis of seven multilocus sequence typing loci were heterogeneous; however, ST29 was the most prevalent ST in the analysed isolates, accounting for 19% (4/21 of the total isolates. Two of the four ST29 isolates had the hypermucoviscosity phenotype. The virulence factors for fimbriae were the most prevalent, followed by siderophores. Community-acquired infections are caused by various species from Klebsiella genus, with different profiles of antibiotic resistance and heterogeneous virulence factors. Keywords: Antimicrobial susceptibility, Bacterial resistance, Cephalosporin resistance, Community infection, ESBL, Hypermucoviscosity

  10. Toxigenic potentiality of Aspergillus flavus and Aspergillus parasiticus strains isolated from black pepper assessed by an LC-MS/MS based multi-mycotoxin method.

    Science.gov (United States)

    Yogendrarajah, Pratheeba; Devlieghere, Frank; Njumbe Ediage, Emmanuel; Jacxsens, Liesbeth; De Meulenaer, Bruno; De Saeger, Sarah

    2015-12-01

    A liquid chromatography triple quadrupole tandem mass spectrometry method was developed and validated to determine mycotoxins, produced by fungal isolates grown on malt extract agar (MEA). All twenty metabolites produced by different fungal species were extracted using acetonitrile/1% formic acid. The developed method was applied to assess the toxigenic potentiality of Aspergillus flavus (n = 11) and Aspergillus parasiticus (n = 6) strains isolated from black peppers (Piper nigrum L.) following their growth at 22, 30 and 37 °C. Highest mean radial colony growth rates were observed at 30 °C for A. flavus (5.21 ± 0.68 mm/day) and A. parasiticus (4.97 ± 0.33 mm/day). All of the A. flavus isolates produced aflatoxin B1 and O-methyl sterigmatocystin (OMST) while 91% produced aflatoxin B2 (AFB2) and 82% of them produced sterigmatocystin (STERIG) at 30 °C. Except one, all the A. parasiticus isolates produced all the four aflatoxins, STERIG and OMST at 30 °C. Remarkably high AFB1 was produced by some A. flavus isolates at 22 °C (max 16-40 mg/kg). Production of mycotoxins followed a different trend than that of growth rate of both species. Notable correlations were found between different secondary metabolites of both species; R(2) 0.87 between AFB1 and AFB2 production. Occurrence of OMST could be used as a predictor for AFB1 production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. isolated from chicken samples.

    Science.gov (United States)

    Er, Buket; Demirhan, Burak; Onurdag, Fatma Kaynak; Ozgacar, Selda Özgen; Oktem, Aysel Bayhan

    2014-03-01

    Salmonella spp. are widespread foodborne pathogens that contaminate egg and poultry meats. Attachment, colonization, as well as biofilm formation capacity of Salmonella spp. on food and contact surfaces of food may cause continuous contamination. Biofilm may play a crucial role in the survival of salmonellae under unfavorable environmental conditions, such as in animal slaughterhouses and processing plants. This could serve as a reservoir compromising food safety and human health. Addition of antimicrobial preservatives extends shelf lives of food products, but even when products are supplemented with adequate amounts of preservatives, it is not always possible to inhibit the microorganisms in a biofilm community. In this study, our aims were i) to determine the minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) of selected preservatives against planktonic and biofilm forms of Salmonella spp. isolated from chicken samples and Salmonella Typhimurium SL1344 standard strain, ii) to show the differences in the susceptibility patterns of same strains versus the planktonic and biofilm forms to the same preservative agent, and iii) to determine and compare antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. For this purpose, Salmonella Typhimurium SL1344 standard strain and 4 Salmonella spp. strains isolated from chicken samples were used. Investigation of antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. was done according to Clinical and Laboratory Standards Institute M100-S18 guidelines and BioTimer assay, respectively. As preservative agents, pure ciprofloxacin, sodium nitrite, potassium sorbate, sodium benzoate, methyl paraben, and propyl paraben were selected. As a result, it was determined that MBIC values are greater than the MIC values of the preservatives. This result verified the resistance seen in a biofilm community to food

  12. Diversity of Cronobacter spp. isolates from the vegetables in the middle-east coastline of China.

    Science.gov (United States)

    Chen, Wanyi; Yang, Jielin; You, Chunping; Liu, Zhenmin

    2016-06-01

    Cronobacter spp. has caused life-threatening neonatal infections mainly resulted from consumption of contaminated powdered infant formula. A total of 102 vegetable samples from retail markets were evaluated for the presence of Cronobacter spp. Thirty-five presumptive Cronobacter isolates were isolated and identified using API 20E and 16S rDNA sequencing analyses. All isolates and type strains were characterized using enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), and genetic profiles of cluster analysis from this molecular typing test clearly showed that there were differences among isolates from different vegetables. A polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) based on the amplification of the gyrB gene (1258 bp) was developed to differentiate among Cronobacter species. A new PCR-RFLP assay based on the amplification of the gyrB gene using Alu I and Hinf I endonuclease combination is established and it has been confirmed an accurate and rapid subtyping method to differentiate Cronobacter species. Sequence analysis of the gyrB gene was proven to be suitable for the phylogenetic analysis of the Cronobacter strains, which has much better resolution based on SNPs in the identification of Cronobacter species specificity than PCR-RFLP and ERIC-PCR. Our study further confirmed that vegetables are one of the most common habitats or sources of Cronobacter spp. contamination in the middle-east coastline of China.

  13. A novel pathway for nicotine degradation by Aspergillus oryzae 112822 isolated from tobacco leaves.

    Science.gov (United States)

    Meng, Xiang Jing; Lu, Li Li; Gu, Guo Feng; Xiao, Min

    2010-09-01

    An efficient nicotine-degrading fungus was isolated from tobacco leaves and identified as Aspergillus oryzae 112822 based on morphological characteristics and sequence analysis of 18S rDNA, 5.8S rDNA and the internal transcribed spacer (5.8S-ITS region). When the strain was cultured in a medium with tobacco leaf extract for 40 h, the maximum amount of cell growth was 3.6 g l(-1) and nicotine degradation was 2.19 g l(-1). The intermediates of nicotine degradation by resting cells were isolated by preparative TLC or semi-preparative HPLC, and identified by TLC, MS, NMR, Fourier-transform (FT)-IR and GC-MS analysis. The pathway for nicotine degradation in A. oryzae 112822 was proposed to be from nicotine to 2,3-dihydroxypyridine through the intermediates nornicotine, myosmine, N-methylnicotinamide and 2-hydroxy-N-methylnicotinamide. The ring of 2,3-dihydroxypyridine was opened between the 2- and 3-hydroxy positions to yield succinic acid. N-methylnicotinamide and 2,3-dihydroxypyridine were satisfactorily verified as metabolites of nicotine degradation. This is the first elucidation of a pathway for nicotine degradation in fungi. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  14. Antimicrobial resistance among Pseudomonas spp. and the Bacillus cereus group isolated from Danish agricultural soil

    DEFF Research Database (Denmark)

    Jensen, Lars Bogø; Baloda, S.; Boye, Mette

    2001-01-01

    From four Danish pig farms, bacteria of Pseudomonas spp. and the Bacillus cereus group were isolated from soil and susceptibility towards selected antimicrobials was tested. From each farm, soil samples representing soil just before and after spread of animal waste and undisturbed agricultural so...... spp., and for bacitracin, erythromycin, penicillin and streptomycin for the B. cereus group. Variations in resistance levels were observed when soil before and after spread of animal waste was compared, indicating an effect from spread of animal waste.......From four Danish pig farms, bacteria of Pseudomonas spp. and the Bacillus cereus group were isolated from soil and susceptibility towards selected antimicrobials was tested. From each farm, soil samples representing soil just before and after spread of animal waste and undisturbed agricultural soil......, when possible, were collected. Soil from a well-characterized Danish farm soil (Hojbakkegaard) was collected for comparison. The Psudomonas spp. and B. cereus were chosen as representative for Gram-negative and Gram-positive indigenous soil bacteria to test the effect of spread of animal waste...

  15. Antimicrobial Resistance of Shigella spp. isolated in the State of Pará, Brazil

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    Flávia Corrêa Bastos

    2011-10-01

    Full Text Available INTRODUCTION: Shigella spp. are Gram-negative, nonsporulating, rod-shaped bacteria that belong to the family Enterobacteriaceae and are responsible for shigellosis or bacillary dysentery, an important cause of worldwide morbidity and mortality. METHODS: We studied the antibiotic resistance profiles of 122 Shigella spp. strains (81 S. flexneri, 41 S. sonnei, 1 S. boydii isolated from patients (female and male from 0 to 80 years of age presenting diarrhea in different districts of the State of Pará, in the North of Brazil. The antibiotic resistance of the strains, isolated from human fecal samples, was determined by the diffusion disk method and by using the VITEK-2 system. RESULTS: The highest resistance rate found was the resistance rate to tetracycline (93.8%, followed by the resistance rate to chloramphenicol (63.9% and to trimethoprim/sulfamethoxazole (63.1%. Resistance to at least three drugs was more common among S. flexneri than S. sonnei (39.5% vs. 10%. Six (4.9% strains were susceptible to all the antibiotics tested. All strains were susceptible to cefotaxime, ceftazidime, ciprofloxacin, nalidixic acid and nitrofurantoin. CONCLUSIONS: High rates of multidrug resistance in Shigella spp. are a serious public health concern in Brazil. It is extremely important to continuously monitor the antimicrobial resistances of Shigella spp. for effective therapy and control measures against shigellosis.

  16. Frecuencia de aislamiento de Staphylococcus spp meticilina resistentes y Enterococcus spp vancomicina resistentes en hospitales de Cuba Frequency of methicilline-resistant Staphylococcus spp and vancomycin-resistant Enterococcus spp isolates in Cuban hospitals

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    Leonora González Mesa

    2005-12-01

    Cuba , there was no updated data either on the rate of infection by methicilline-resistant Staphylococcus spp or on the circulation of this germ in the community; neither are there reports on vancomycin-resistant Enterococcus spp presence. In this study, 774 strains collected from hospitals in the country were analyzed. The mechanism of resistance was determined by the methods suggested in the NCCLS guidelines. The 9.3 % (23 and 4.0 % (7 of S. aureus isolates from the hospitals and the community respectively were methicilline-resistant carriers of mecA gen whereas 69.9 %(72 of negative Staphylococcus coagulase isolates showed resistance to oxacillin. Also, a vancomycin-resistant Enterococcus spp-carrying strain was detected. Our results revealed that in Cuba the methicilline-resistant S. aureus is not a problem neither at hospitals nor at the community setting. Despite the fact that the circulation of these germs in the community setting and also the circulation of vancomycin-resistant Enterococcus spp at hospital setting have been reported for the first time, their frequency is very low as a consequence of the advances in the implementation of policies aimed at a more rational use and consumption of antibiotics.

  17. Prevalence of Salmonella spp., and serovars isolated from captive exotic reptiles in New Zealand.

    Science.gov (United States)

    Kikillus, K H; Gartrell, B D; Motion, E

    2011-07-01

    To investigate the prevalence of Salmonella spp. in captive exotic reptile species in New Zealand, and identify the serovars isolated from this population. Cloacal swabs were obtained from 378 captive exotic reptiles, representing 24 species, residing in 25 collections throughout New Zealand between 2008 and 2009. Samples were cultured for Salmonella spp., and suspected colonies were serotyped by the Institute of Environmental Science and Research (ESR). Forty-three of the 378 (11.4%) reptiles sampled tested positive for Salmonella spp., with 95% CI for the estimated true prevalence being 12-25% in exotic reptiles in this study population. Lizards tested positive for Salmonella spp. more often than chelonians. Agamid lizards tested positive more often than any other family group, with 95% CI for the estimated true prevalence being 56-100%.. Six Salmonella serovars from subspecies I and two from subspecies II were isolated. The serovar most commonly isolated was S. Onderstepoort (30.2%), followed by S. Thompson (20.9%), S. Potsdam (14%), S. Wangata (14%), S. Infantis (11.6%) and S. Eastbourne (2.3%). All of the subspecies I serovars have been previously reported in both reptiles and humans in New Zealand, and include serovars previously associated with disease in humans. This study showed that Salmonella spp. were commonly carried by exotic reptiles in the study population in New Zealand. Several serovars of Salmonella spp. with known pathogenicity to humans were isolated, including S. Infantis, which is one of the most common serovars isolated from both humans and non-human sources in New Zealand. The limitations of this study included the bias engendered by the need for voluntary involvement in the study, and the non-random sampling design. Based on the serovars identified in this and previous studies, it is recommended native and exotic reptiles be segregated within collections, especially when native reptiles may be used for biodiversity restoration

  18. Genetic characterization of Shigella spp. isolated from diarrhoeal and asymptomatic children.

    Science.gov (United States)

    Ghosh, Santanu; Pazhani, Gururaja P; Niyogi, Swapan Kumar; Nataro, James P; Ramamurthy, Thandavarayan

    2014-07-01

    Phenotypic and genetic characteristics of Shigella spp. isolated from diarrhoeal and asymptomatic children aged up to 5 years were analysed in this study. In total, 91 and 17 isolates were identified from diarrhoeal (case) and asymptomatic (control) children, respectively. All the isolates were tested for antimicrobial resistance, the presence of integrons, plasmid-mediated quinolone resistance (PMQR), virulence-associated genes and Shigella pathogenicity island (SH-PAI). The majority of the Shigella spp. from cases (68.1%) and controls (82.3%) were found to be resistant to fluoroquinolones. Integron carriage was detected more in cases (76.9%) than in controls (35.5%). Atypical class 1 integron was detected exclusively in Shigella flexneri from cases but not from the controls. PMQR genes such as aac(6')-Ib-cr and qnrS1 were detected in 82.4 and 14.3% of the isolates from cases and in 53 and 17.6% in controls, respectively. Shigella isolates from cases as well as from controls were positive for the invasive plasmid antigen H-encoding gene ipaH. The other virulence genes such as virF, sat, setA, setB, sen and ial were detected in Shigella isolates in 80.2, 49.4, 27.4, 27.4, 80.2 and 79.1% of cases and in 64.7, 52.9, 17.6, 17.6, 64.7 and 64.7% of controls, respectively. The entire SH-PAI was detected in S. flexneri serotype 2a from cases and controls. In an isolate from a control child, the SH-PAI was truncated. Integrons, PMQR and virulence-encoding genes were detected more frequently in cases than in controls. In diarrhoea endemic areas, asymptomatic carriers may play a crucial role in the transmission of multidrug-resistant Shigella spp. with all the putative virulence genes. © 2014 The Authors.

  19. Diversity of black Aspergilli isolated from raisins in Argentina: Polyphasic approach to species identification and development of SCAR markers for Aspergillus ibericus.

    Science.gov (United States)

    Giaj Merlera, G; Muñoz, S; Coelho, I; Cavaglieri, L R; Torres, A M; Reynoso, M M

    2015-10-01

    Aspergillus section Nigri is a heterogeneous fungal group including some ochratoxin A producer species that usually contaminate raisins. The section contains the Series Carbonaria which includes the toxigenic species Aspergillus carbonarius and nontoxigenic Aspergillus ibericus that are phenotypically undistinguishable. The aim of this study was to examine the diversity of black aspergilli isolated from raisins and to develop a specific genetic marker to distinguish A. ibericus from A. carbonarius. The species most frequently found in raisins in this study were Aspergillus tubingensis (35.4%) and A. carbonarius (32.3%), followed by Aspergillus luchuensis (10.7%), Aspergillus japonicus (7.7%), Aspergillus niger (6.2%), Aspergillus welwitschiae (4.6%) and A. ibericus (3.1%). Based on inter-simple sequence repeat (ISSR) fingerprinting profiles of major Aspergillus section Nigri members, a sequence-characterized amplified region (SCAR) marker was identified. Primers were designed based on the conserved regions of the SCAR marker and were utilized in a PCR for simultaneous identification of A. carbonarius and A. ibericus. The detection level of the SCAR-PCR was found to be 0.01 ng of purified DNA. The present SCAR-PCR is rapid and less cumbersome than conventional identification techniques and could be a supplementary strategy and a reliable tool for high-throughput sample analysis. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Clinical characteristics of patients with Aspergillus species isolation from respiratory samples: Comparison of chronic pulmonary aspergillosis and colonization.

    Science.gov (United States)

    Ohara, Sayaka; Tazawa, Yoko; Tanai, Chiharu; Tanaka, Yoshiaki; Noda, Hiromichi; Horiuchi, Hajime; Usui, Kazuhiro

    2016-03-01

    With advancements in anti-fungal drugs, it has become more important to correctly diagnose chronic pulmonary aspergillosis (CPA); however, it is not easy to distinguish CPA from colonization when Aspergillus species are isolated from respiratory samples. The aim of the study was to clarify the particular clinical characteristics of patients with CPA vs. those with colonization. We retrospectively reviewed the medical records of 110 patients with Aspergillus species isolation from respiratory samples, to analyze and compare the differences between CPA and colonization of the Aspergillus species. The median age of all analyzed was 71 years (range: 31-92 years); 64 were female (58%). The most frequently cultured Aspergillus species was Aspergillus fumigatus (48.3%), followed by A. niger (29.2%). Thirty patients (27.4%) were diagnosed with CPA, vs. 75 (68.2%) with colonization and 5 (4.5%) with allergic bronchopulmonary aspergillosis. Compared with the colonization group, the CPA group included more males (CPA vs. colonization: 49.3% vs. 13.3%) and subjects with a low body mass index (18.45 kg/m2 vs. 21.09 kg/m2). As for the underlying pulmonary diseases, the patients with CPA showed a significantly higher prevalence of sequelae of pulmonary tuberculosis (40% vs. 8%) and a history of thoracic surgery (43% vs. 13%) than those with colonization. Asthma was less frequent in the CPA group than in the colonization group (0% vs. 20%). We found no significantly important underlying extrapulmonary diseases. Patients with CPA display clinical characteristics distinct from those seen in subjects with colonization. Copyright © 2015 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.

  1. Antifungal activity of lemon, eucalyptus, thyme, oregano, sage and lavender essential oils against Aspergillus niger and Aspergillus tubingensis isolated from grapes

    Directory of Open Access Journals (Sweden)

    Miroslava Císarová

    2016-01-01

    Full Text Available Today, it is very important to find out the protection of products of natural origin as an alternative to synthetic fungicides. The promising alternative is the use of the essential oils (EOs. Essential oils from plants have great potential as a new source of fungicide to control the pathogenic fungi.The main objective of this study was evaluation of the antifungal activity of lemon (Citrus lemon L., eucalyptus (Eucalyptus globulus LABILL., thyme (Thymus vulgaris L., oregano (Origanum vulgare L. sage (Salvia officinalis L. and lavender (Lavandula angustifolia MILLER. EOs against Aspergillus niger and Aspergillus tubingensis isolated from grapes and their ability to affect the growth. It was tested by using the vapor contact with them. At first both tested isolates were identified by using PCR method. Sequence data of 18S rRNA supported the assignment of these isolates to the genus Aspergillus and species A. niger (ITS region: KT824061; RPB2: KT824060 and A. tubingensis (ITS region: KT824062; RPB2: KT824059. Second, EO antifungal activity was evaluated. The effect of the EO volatile phase was confirmed to inhibit growth of A. niger and A tubingensis. EOs were diluted in DMSO (dimethyl sulfoxide final volume of 100 μL. Only 50 μL this solution was distributed on a round sterile filter paper (1 x 1 cm by micropipette, and the paper was placed in the center of the lid of Petri dishes. Dishes were kept in an inverted position. The essential oils with the most significant activity were determined by method of graded concentration of oils - minimum inhibitory doses (MIDs. The most effective tested EOs were oregano and thyme oils, which totally inhibited growth of tested isolates for all days of incubation at 0.625 μL.cm-3 (in air with MFDs 0.125 μL.cm-3 (in air. Lavender EO was less active aginst tested strains (MIDs 0.313 μL.cm-3. The results showed that the tested EOs had antifungal activity, except lemon and eucalyptus. Sage EO was the only

  2. Dissimilatory nitrate reduction by Aspergillus terreus isolated from the seasonal oxygen minimum zone in the Arabian Sea

    OpenAIRE

    Stief, Peter; Fuchs-Ocklenburg, Silvia; Kamp, Anja; Manohar, Cathrine-Sumathi; Houbraken, Jos; Boekhout, Teun; de Beer, Dirk; Stoeck, Thorsten

    2014-01-01

    Background A wealth of microbial eukaryotes is adapted to life in oxygen-deficient marine environments. Evidence is accumulating that some of these eukaryotes survive anoxia by employing dissimilatory nitrate reduction, a strategy that otherwise is widespread in prokaryotes. Here, we report on the anaerobic nitrate metabolism of the fungus Aspergillus terreus (isolate An-4) that was obtained from sediment in the seasonal oxygen minimum zone in the Arabian Sea, a globally important site of oce...

  3. Species identification of Aspergillus section Flavi isolates from Portuguese almonds using phenotypic, including MALDI-TOF ICMS, and molecular approaches

    OpenAIRE

    Rodrigues, Paula; Venâncio, Armando; Lima, Nelson

    2011-01-01

    Section Flavi is one of the most significant Sections in the genus Aspergillus. Taxonomy of this section currently depends on multivariate approaches, entailing phenotypic and molecular traits. This work aimed to identify isolates from section Flavi by combining various classic phenotypic and genotypic methods as well as the novel approach based on spectral analysis by MALDI-TOF ICMS, and to evaluate the discriminatory power of the various approaches in species identification. Methods and ...

  4. Effect of Antimicrobials on Salmonella Spp. Strains Isolated from Poultry Processing Plants

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    L Mion

    Full Text Available ABSTRACT The routine use of antimicrobials in animal production for the treatment of infections, disease prevention, or as growth promoters is a predisposing factor for the development and dissemination of antimicrobial resistance. In food industries, sanitizers are used for the control of microbial colonization, and their efficacy depends on contact time and on the dilution of the products used. The present study assessed the effect of 12 antimicrobials and four commercial sanitizers on 18 Salmonella spp. strains isolated from poultry processing plants. None of the evaluated antimicrobials was 100% effective against the tested Salmonella spp. strains; however, 94% of the isolates were susceptible to ciprofloxacin, 77% to amoxicillin + clavulanic acid and to ampicillin, and 72% to enrofloxacin, whereas 100% of the isolates were resistant to penicillin G, 16% to tetracycline, and 11% to sulfonamide. The tested Salmonella spp. strains were 100% inhibited by peracetic acid after five minutes of contact, 0.5% by quaternary ammonium after 15 minutes, and 85.7% by chlorhexidine after 15 minutes. The results indicate the importance of testing of efficacy of antimicrobials used in animal production and in public health to monitor their action and the development of resistance.

  5. Keratin Production by Decomposing Feather Waste Using Some Local Bacillus spp. Isolated from Poultry Soil

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    Mojtaba Salouti

    2016-12-01

    Full Text Available Background: Feather waste is generated in large amounts as a by-product of commercial poultry processing. The main component of feather is keratin. The main purpose of this study was to identify Bacillus spp. (the keratinolytic bacteria that are able to degrade the feather for producing keratin. Methods: Bacillus spp. Were isolated from the waste of poultries located in Miyaneh city. The bacteria were grown on basal medium containing 1% hen feather as the sole source of carbon ,nitrogen, sulfur and energy at 27ºC for 7 days. Then,the isolates capable of feather degrading were identified. The Bradford method was used to assay the production of keratin in the feather samples. Different pH and temperatures were studied to determine the best conditions for production of keratinase enzyme. Results: Seven Bacillus spp. including: B. pumilis, B. subtilis, B. firmus, B. macerance, B. popilliae, B. lentimorbus and B. larvae were found to be able to degrade the feather with different abilities. Conclusion: B. subtilis was found to be most productive isolate for keratinase enzyme production.

  6. Molecular epidemiology and virulence assessment of Aspergillus fumigatus isolates from white stork chicks and their environment.

    Science.gov (United States)

    Olias, Philipp; Gruber, Achim D; Hafez, Hafez M; Lierz, Michael; Slesiona, Silvia; Brock, Matthias; Jacobsen, Ilse D

    2011-03-24

    Aspergillus fumigatus is a common pathogen in poultry and captive wild birds and an emerging opportunistic fungal pathogen in immunocompromised humans. Although invasive aspergillosis is frequently reported in free-ranging wild birds, the incidence and epidemiology of the disease in a natural setting is unknown. We recently reported endemic outbreaks of invasive aspergillosis at white stork nesting sites close to human habitation in Germany with significant subsequent breeding losses. Therefore, we hypothesized that A. fumigatus strains with higher virulence in birds may have evolved in this environment and performed the first epidemiological analysis of invasive aspergillosis in free-ranging wild birds. Sixty-one clinical and environmental A. fumigatus isolates from six affected nesting sites were genotyped by microsatellite analysis using the STRAf-assay. The isolates showed a remarkable high genomic diversity and, contrary to the initial hypothesis, clinical and environmental isolates did not cluster significantly. Interestingly, storks were infected with two to four different genotypes and in most cases both mating types MAT-1.1 and MAT-1.2 were present within the same specimen. The majority of selected clinical and environmental strains exhibited similar virulence in an in vivo infection model using embryonated chicken eggs. Noteworthy, virulence was not associated with one distinct fungal mating type. These results further support the assumption that the majority of A. fumigatus strains have the potential to cause disease in susceptible hosts. In white storks, immaturity of the immune system during the first three weeks of age may enhance susceptibility to invasive aspergillosis. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Isolation, identification of an axenic fungal isolate of aspergillus sp. (mbl-1511) and its subsequent improvement for enhanced extracellular lipolytic potential through monoculture fermentation

    International Nuclear Information System (INIS)

    Iftikhar, T.; Sidra, A.; Ali, M.; Majeed, H.; Abdullah, R.

    2017-01-01

    The present investigation was conducted for extracellular lipases production. One hundred and forty samples of fungi were isolated from different environment and food samples. Among all the isolated cultures, an isolate obtained from chicken roasted in oil (MBL-1511) gave the highest extracellular lipase through SSF. Hyper producer strain (MBL-1511) was morphologically identified. A morphologically identified isolate of Aspergillus niger (MBL 1511) was verified by DNA barcoding marker like 18S rRNA gene sequence. The sequence of Aspergillus niger (MBL 1511) was registered with accession no. [GenBank: KP172477] in the public nucleotide library (genbank) of NCBI. The selected hyper producer of Aspergillus niger (MBL-1511) strain was subjected to physical and chemical mutagenic treatments to improve its lipolytic potential. Proximate analysis confirmed brassica meal as the best basal substrate with the lipases potential of 10.67+-0.01 IU/mL (wild) and 19.58+-0.04 IU/mL (mutant). The optimum conditions for the maximized extracellular lipases production were 1.0 mL inoculum at 30 degree C after 72 h at pH of 6.2. Finally, a potent mutant of A. niger [MBL-1511SA-4(150 min)] with an increased activity of 161 % over the wild strain was obtained when olive oil was used at 1% (v/v) concentration. (author)

  8. Identification of morphological and molecular Aspergillus species isolated from patients based on beta-tubulin gene sequencing

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    Mahnaz Kheirkhah

    2017-06-01

    Full Text Available Background: Aspergillus species are opportunistic pathogens among immunocompromised patients. In terms of pathogenesis and mycotoxin production, they are in great value. The aim of the this study was to evaluate of beta-tubulin gene for identification of clinical Aspergillus species by PCR-sequencing method compared to morphological features of clinical isolates (such as conidial shape in direct microscopic examination, colony shape in culture, and physiological tests. Materials and Methods: In this study, 465 patients referred to the Shefa laboratory of Isfahan were evaluated. Morphological and molecular identification of clinical samples were performed using culture on sabouraud agar, malt extract agar, czapekdox agar, direct microscopy, and PCR-sequencing of beta tubulin gene, respectively. Sequences were analyzed in comparison with gene bank data. Results: Thirty nine out of 465 suspected cases (8.4% had aspergillosis. The most prevalent species were Aspergillus flavus (56.4%, A. oryzae (20.5%, and A. fumigatus (10.2%, respectively. Fifty nine percent of patients were females and 49% were males. Conclusion: In comparison with phenotypic tests, sequencing of beta-tubulin gene for identification of Aspergillus species is at great value. Replacement of molecular techniques with conventional tests is recommended for precise identification of microorganism for better management of infection.

  9. Urease-positive thermophilic strains of Campylobacter isolated from seagulls (Larus spp.).

    Science.gov (United States)

    Kaneko, A; Matsuda, M; Miyajima, M; Moore, J E; Murphy, P G

    1999-07-01

    Three strains of urease-positive thermophilic Campylobacter (UPTC), designated A1, A2 and A3, were identified by biochemical characterization after isolation from faeces of seagulls in Northern Ireland in 1996. The biochemical characteristics of the strains were identical to those of strains described previously. Analysis by pulsed-field gel electrophoresis (PFGE) after separate digestion with ApaI and SmaI demonstrated that the respective PFGE profiles were indistinguishable. The PFGE analysis also suggested that the genomes were approximately 1810 kb in length. This is the first example of the isolation of UPTC from flying homoiothermal animals, i.e. from seagulls (Larus spp.).

  10. Antibiotic Resistance of Salmonella spp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil

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    Fátima C. T. Carvalho

    2013-01-01

    Full Text Available This study investigated the presence and antibiotic resistance of Salmonella spp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimated Litopenaeus vannamei were examined for the presence of Salmonella. Afterwards, Salmonella isolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12% of the 186 isolates were confirmed to be Salmonella spp., belonging to five serovars: S. serovar Saintpaul, S. serovar Infantis, S. serovar Panama, S. serovar Madelia, and S. serovar Braenderup, along with 2 subspecies: S. enterica serovar houtenae and S. enterica serovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area.

  11. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

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    Ibrahim Che Omar

    2005-03-01

    Full Text Available Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF using palm kernel cake (PKC as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC. The supplementation of additional carbon and nitrogen sources in the PKC medium could enhance enzyme productivity. The maximum production of xylanase and growth obtained with the supplementation of xylose at 0.75% (w/w were 25.40 U/g and 1.69 mg glucosamine/ g PKC. Moreover, the presence of NaNO3 at 0.075% (w/w as additional nitrogen source further enhanced xylanase production to 33.99 U/g PKC although the growth remained unchanged at about 1.67 mg glucosa- mine/g PKC. The optimized conditions showed an increased xylanase production by 157% compared to before the optimization of the SSF system. The xylanase productivity was 23.12 U/mg glucosamine after optimization and 11.72 U/mg glucosamine before optimization.

  12. Bioleaching of nickel by Aspergillus humicola SKP102 isolated from Indian lateritic overburden

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    Suchhanda Ghosh

    2016-01-01

    Full Text Available The lateritic deposits spread over the Eastern Ghats of Sukinda Valley, Odisha, India, produce a huge amount of overburden annually as a byproduct of chromite mining. This chromite mining overburden contains nickel, the only source of the metal in the country. During this study Aspergillus humicola SKP102, an indigenous fungus isolated from the mining overburden was employed for the leaching of nickel. About 53.89% of the nickel could be leached by the fungus when grown in batch mode using a Czapek dox medium containing 2% (w/v of the mining overburden. The parameters affecting bioleaching were optimized in order to grow the fungus and leach the metal. Of the different options of cheap carbon sources, straw infusion and molasses emerged as viable options for the growth of the fungus and the leaching of nickel. Two-step and indirect techniques were also used for this purpose, and they resulted in 53.09% and 65.04% Ni leaching respectively. Adding diluted sulfuric acid to the leaching medium resulted in 97.05% nickel recovery from the overburden pulp. A. humicola SKP102 could be a potential tool for leaching nickel from the mining overburden.

  13. Rapid Isolation and Susceptibility Testing of Leptospira spp. Using a New Solid Medium, LVW Agar

    Science.gov (United States)

    Wuthiekanun, Vanaporn; Amornchai, Premjit; Paris, Daniel H.; Langla, Sayan; Thaipadunpanit, Janjira; Chierakul, Wirongrong; Smythe, Lee D.; White, Nicholas J.; Day, Nicholas P. J.; Peacock, Sharon J.

    2013-01-01

    Pathogenic Leptospira spp., the causative agents of leptospirosis, are slow-growing Gram-negative spirochetes. Isolation of Leptospira from clinical samples and testing of antimicrobial susceptibility are difficult and time-consuming. Here, we describe the development of a new solid medium that facilitates more-rapid growth of Leptospira spp. and the use of this medium to evaluate the Etest's performance in determining antimicrobial MICs to drugs in common use for leptospirosis. The medium was developed by evaluating the effects of numerous factors on the growth rate of Leptospira interrogans strain NR-20157. These included the type of base agar, the concentration of rabbit serum (RS), and the concentration and duration of CO2 incubation during the initial period of culture. The highest growth rate of NR-20157 was achieved using a Noble agar base supplemented with 10% RS (named LVW agar), with an initial incubation at 30°C in 5% CO2 for 2 days prior to continuous culture in air at 30°C. These conditions were used to develop the Etest for three species, L. interrogans (NR-20161), L. kirschnerii (NR-20327), and L. borgpetersenii (NR-20151). The MICs were read on day 7 for all samples. The Etest was then performed on 109 isolates of pathogenic Leptospira spp. The MIC90 values for penicillin G, doxycycline, cefotaxime, ceftriaxone, and chloramphenicol were 0.64 units/ml and 0.19, 0.047, 0.5, and 2 μg/ml, respectively. The use of LVW agar, which enables rapid growth, isolation of single colonies, and simple antimicrobial susceptibility testing for Leptospira spp., provides an opportunity for new areas of fundamental and applied research. PMID:23114772

  14. Field study on the epidemiology and pathogenicity of different isolates of bovine Ostertagia spp.

    Science.gov (United States)

    Al Saqur, I; Armour, J; Bairden, K; Dunn, A M; Jennings, F W; Murray, M

    1982-11-01

    The epidemiological features of three different isolates of bovine Ostertagia spp under similar initial levels of larval challenge were compared in the field. Two of the isolates, consisting mainly of Ostertagia ostertagi, and a low proportion of Skrjabinagia lyrata conformed in epidemiological behaviour with those investigated by previous workers, though the worm burdens which established did not give rise to the expected clinical signs. The third isolate behaved in a different way, yielding very high faecal egg counts which were followed by high pasture larval counts, heavy worm burdens and severe clinical disease. This isolate, while consisting mainly of O ostertagi and a few S lyrata, also contained a proportion of O leptospicularis, and it is suggested that this species may influence the dynamics of the host-parasite relationship in bovine ostertagiasis.

  15. Isolation, identification, and characterization of Listeria spp. from various animal origin foods

    Science.gov (United States)

    Nayak, Deepti N.; Savalia, C. V.; Kalyani, I. H.; Kumar, Rajeev; Kshirsagar, D. P.

    2015-01-01

    Aim: The present study was undertaken with the prime objective of isolating and identifying Listeria spp. from various foods of animal origin sold at retail market outlets in the city of Navsari, Gujarat. Materials and Methods: Total 200 samples comprising of milk, milk products, meat, and fish (50 each) collected aseptically from local market which were subjected first to pre-enrichment in half strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on PALCAM agar. The growth with the typical colony characteristics were further identified up to species level on the basis of their morphological and biochemical characteristics. Cultures identified as Listeria monocytogenes were further subjected to in vitro pathogenicity tests and detection of different virulence-associated genes viz. actA, hlyA, and iap using polymerase chain reaction. Results: Of the total 200 food samples of animal origin; 18 (9%) were found positive for Listeria spp. which were identified as Listeria seeligeri (6, 33.3%), Listeria innocua (5, 27.7%), Listeria welshimeri (4, 22.2%), and L. monocytogenes (3, 16.6%). The highest prevalence was observed in milk samples (8). Species wise, 6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk). All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iap using the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers. Conclusion: Listeria spp. was isolated from 9% (18/200) of the animal origin food samples viz.; milk, milk products, meat, and fish with the highest prevalence

  16. Isolation, identification, and characterization of Listeria spp. from various animal origin foods

    Directory of Open Access Journals (Sweden)

    Deepti N. Nayak

    2015-06-01

    Full Text Available Aim: The present study was undertaken with the prime objective of isolating and identifying Listeria spp. from various foods of animal origin sold at retail market outlets in the city of Navsari, Gujarat. Materials and Methods: Total 200 samples comprising of milk, milk products, meat, and fish (50 each collected aseptically from local market which were subjected first to pre-enrichment in half strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on PALCAM agar. The growth with the typical colony characteristics were further identified up to species level on the basis of their morphological and biochemical characteristics. Cultures identified as Listeria monocytogenes were further subjected to in vitro pathogenicity tests and detection of different virulence associated genes viz. actA, hlyA, and iap using polymerase chain reaction. Results: Of the total 200 food samples of animal origin; 18 (9% were found positive for Listeria spp. which were identified as Listeria seeligeri (6, 33.3%, Listeria innocua (5, 27.7%, Listeria welshimeri (4, 22.2%, and L. monocytogenes (3, 16.6%. The highest prevalence was observed in milk samples (8. Species wise, 6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk. All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iap using the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers. Conclusion: Listeria spp. was isolated from 9% (18/200 of the animal origin food samples viz.; milk, milk products, meat, and fish with the highest

  17. An attempt to optimize potassium sorbate use to preserve low pH (4.5-5.5) intermediate moisture bakery products by modelling Eurotium spp., Aspergillus spp. and Penicillium corylophilum growth.

    Science.gov (United States)

    Guynot, M Elena; Marín, Sonia; Sanchis, Vicente; Ramos, Antonio J

    2005-05-25

    Mould growth was modelled on fermented bakery product analogues (FBPA) of two different pH (4.5 and 5.5), different water activity (a(w)) levels (0.80-0.90) and potassium sorbate concentrations (0-0.3%) by using seven moulds commonly causing spoilage of bakery products (Eurotium spp., Aspergillus spp. and Penicillium corylophilum). For the description of fungal growth (growth rates) as a function of a(w), potassium sorbate concentration and pH, 10-terms polynomial models were developed. Modelling enables prediction of spoilage during storage as a function of the factors affecting fungal growth. At pH 4.5 the concentration of potassium sorbate could be reduced to some extent only at low levels of a(w), whereas at pH 5.5 fungal growth was observed even by adding 0.3% of potassium sorbate. However, this preservative could be a valuable alternative as antifungal in such bakery product, of slightly acidic pH, if a long shelf life has not to be achieved.

  18. Removal of Heavy Metals from Synthesis Industrial Wastewater Using Local Isolated Candida Utilis and Aspergillus Niger as Bio-Filter

    OpenAIRE

    Safaa Abd Alrasool Ali

    2013-01-01

    In this study biomass filter of Candida Utilis which isolated from food sample in Baghdad local market and Aspergillus Niger which isolated from Baghdad soil used to biosorption heavy metals from synthesis industrial wastewater. two bio-filters were designed as cylindrical Perspex with height 10cm, diameter 3cm as a shell and inside their are layers of Sponge were prepare as the size of diameter of the cylindrical filter with thickness of 2cm arrange inside it, biomass production were 6 g of ...

  19. Molecular and phylogenetic characterisation of Fasciola spp. isolated from cattle and sheep in southeastern Iran

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    H. Mirahmadi

    2018-03-01

    Full Text Available The objective of this study was to determine the genotype of Fasciola spp. in animal hosts from Za-hedan, Sistan and Baluchestan province, southeastern Iran using PCR-RFLP. Overall, 50 and 43 adult Fasciola spp. were isolated from bile ducts of naturally infected cattle and sheep. PCR-RFLP with RsaI restriction enzyme and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS 1 region from Fasciola spp. were used to conduct the study. RFLP pattern with RsaI produced 180 and 331 bp fragments in F. gigantica and amplicons of F. hepatica had a size of 77, 104 and 331 bp. Results based on PCR-RFLP analysis were confirmed by sequence analysis of repre-sentative ITS 1 amplicons. No hybrid forms were detected in the present study. All sheep were in-fected with F. hepatica while cattle were infected with both species. The results of our study showed that F. hepatica and F. gigantica isolates were of common H1 and G1 haplotypes.

  20. Molecular study on some antibiotic resistant genes in Salmonella spp. isolates

    Science.gov (United States)

    Nabi, Ari Q.

    2017-09-01

    Studying the genes related with antimicrobial resistance in Salmonella spp. is a crucial step toward a correct and faster treatment of infections caused by the pathogen. In this work Integron mediated antibiotic resistant gene IntI1 (Class I Integrase IntI1) and some plasmid mediated antibiotic resistance genes (Qnr) were scanned among the isolated non-Typhoid Salmonellae strains with known resistance to some important antimicrobial drugs using Sybr Green real time PCR. The aim of the study was to correlate the multiple antibiotics and antimicrobial resistance of Salmonella spp. with the presence of integrase (IntI1) gene and plasmid mediated quinolone resistant genes. Results revealed the presence of Class I Integrase gene in 76% of the isolates with confirmed multiple antibiotic resistances. Moreover, about 32% of the multiple antibiotic resistant serotypes showed a positive R-PCR for plasmid mediated qnrA gene encoding for nalidixic acid and ciprofloxacin resistance. No positive results could be revealed form R-PCRs targeting qnrB or qnrS. In light of these results we can conclude that the presence of at least one of the qnr genes and/or the presence of Integrase Class I gene were responsible for the multiple antibiotic resistance to for nalidixic acid and ciprofloxacin from the studied Salmonella spp. and further studies required to identify the genes related with multiple antibiotic resistance of the pathogen.

  1. Antagonistic Bioactivity of Endophytic Actinomycetes Isolated from Medicinal Plants

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    M. Gangwar

    2011-10-01

    Full Text Available Endophytic actinomycetes are promising biocontrol agents for use in agriculture and have been isolated from various plant species. In the present study, 40 endophytic actinomycetes were isolated from roots, stems and leaves of three medicinal plants viz. Aloe vera, Mentha arvensis and Ocimum sanctum. The identification revealed that the majority of the isolates were Streptomyces spp. and the rest were identified as Saccharopolyspora spp., Micromonospora spp. and Actinopolyspora spp. The dual tests revealed that nine endophytic actinomycete isolates displayed a wide spectrum activity against nine fungal phytopathogens. Out of 8 isolates, 90% inhibited the growth of at least one or more phytopathogenic fungi and Saccharopolyspora 0-9 (Out of 8 isolates, 90% inhibited the growth of at least one or more phytopathogenic fungi and Saccharopolyspora 0-9 exhibited antagonistic activity against Aspergillus niger, Aspergillus flavus, Alternaria brassicicola, Botrytis cinerea, Penicillium digitatum, Fusarium oxysporum, Penicillium pinophilum, Phytophthora dresclea and Colletotrichum falcatum.

  2. Short communication: β-Lactam resistance and vancomycin heteroresistance in Staphylococcus spp. isolated from bovine subclinical mastitis.

    Science.gov (United States)

    Mello, Priscila Luiza; Pinheiro, Luiza; Martins, Lisiane de Almeida; Brito, Maria Aparecida Vasconcelos Paiva; Ribeiro de Souza da Cunha, Maria de Lourdes

    2017-08-01

    The use of antimicrobial agents has led to the emergence of resistant bacterial strains over a relatively short period. Furthermore, Staphylococcus spp. can produce β-lactamase, which explains the survival of these strains in a focus of infection despite the use of a β-lactam antibiotic. The aim of this study was to evaluate the resistance of Staphylococcus spp. isolated from bovine subclinical mastitis to oxacillin and vancomycin (by minimum inhibitory concentration) and to detect vancomycin heteroresistance by a screening method. We also evaluated β-lactamase production and resistance due to hyperproduction of this enzyme and investigated the mecA and mecC genes and performed staphylococcal cassette chromosome mec typing. For this purpose, 181 Staphylococcus spp. isolated from mastitis subclinical bovine were analyzed. Using the phenotypic method, 33 (18.2%) of Staphylococcus spp. were resistant to oxacillin. In contrast, all isolates were susceptible to vancomycin, and heteroresistance was detected by the screening method in 13 isolates. Production of β-lactamase was observed in 174 (96%) of the Staphylococcus spp. isolates. The mecA gene was detected in 8 isolates, all of them belonging to the species Staphylococcus epidermidis, and staphylococcal cassette chromosome mec typing revealed the presence of type I and type IV isolates. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  3. Isolation, molecular identification and quinolone-susceptibility testing of Arcobacter spp. isolated from fresh vegetables in Spain.

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    González, Ana; Bayas Morejón, Isidro Favián; Ferrús, María Antonia

    2017-08-01

    Some species of the Arcobacter genus are considered emerging foodborne and waterborne enteropathogens. However, the presence of Arcobacter spp. in vegetables very little is known, because most studies have focused on foods of animal origin. On the other hand, quinolones are considered as first-line drugs for the treatment of infection by campylobacteria in human patients, but few data are currently available about the resistance levels to these antibiotics among Arcobacter species. Therefore, the aim of this study was to investigate the presence and diversity of arcobacters isolated from fresh vegetables such as lettuces, spinaches, chards and cabbages. Resistance to quinolones of the isolates was also investigated. One hundred fresh vegetables samples purchased from seven local retail markets in Valencia (Spain) during eight months were analysed. The study included 41 lettuces, 21 spinaches, 34 chards and 4 cabbages. Samples were analysed by culture and by molecular methods before and after enrichment. By culture, 17 out of 100 analysed samples were Arcobacter positive and twenty-five isolates were obtained from them. Direct detection by PCR was low, with only 4% Arcobacter spp. positive samples. This percentage increased considerably, up 20%, after 48 h enrichment. By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), 17 out of the 25 isolates were identified as A. butzleri and 8 as A. cryaerophilus. Only two A. butzleri isolates showed resistance to levofloxacin and ciprofloxacin. The sequencing of a fragment of the QRDR region of the gyrA gene from the quinolones-resistant isolates revealed the presence of a mutation in position 254 of this gene (C-T transition). This study is the first report about the presence of pathogenic species of Arcobacter spp. in chards and cabbages and confirms that fresh vegetables can act as transmission vehicle to humans. Moreover, the presence of A. butzleri quinolone resistant in vegetables could

  4. Characterization of Treponema spp. isolates from pigs with ear necrosis and shoulder ulcers.

    Science.gov (United States)

    Svartström, Olov; Karlsson, Frida; Fellström, Claes; Pringle, Märit

    2013-10-25

    Ear necrosis and shoulder ulcers in pigs are animal welfare problems and ethical issues that can cause economic losses for producers. Spirochetes have been observed microscopically in scrapings from pig ulcers since the early 1900s, but have until recently not been cultured and therefore not characterized. In this study, 12 Treponema spp. isolates were acquired from porcine ear necrosis, shoulder ulcers and gingiva. DNA analysis of the 16S rRNA-tRNA(Ile) intergenic spacer region (ISR2) or the 16S rRNA gene revealed relatedness to oral treponemes found in dogs and humans. All isolates except one aligned into two clusters, Treponema pedis and Treponema sp. OMZ 840-like. The 16S rRNA gene of the remaining isolate shared 99% nucleotide identity with Treponema parvum. Genetic fingerprinting of the isolates was performed through random amplification of polymorphic DNA (RAPD). In addition, the isolates were characterized by biochemical tests, including api(®)ZYM, tryptophanase and hippuricase activity, and by testing the antimicrobial susceptibility to tiamulin, valnemulin, tylosin, tylvalosin, lincomycin and doxycycline using broth dilution. All isolates except two showed unique RAPD fingerprints, whereas metabolic activity tests could not differentiate between the isolates. The MICs of all antimicrobial agents tested were low. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Prevalence of Candida spp. in cervical-vaginal samples and the in vitro susceptibility of isolates

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    Tchana Martinez Brandolt

    Full Text Available Abstract Vulvovaginal candidiasis (VVC is an infection of the genital mucosa caused by different species of the genus Candida. Considering the lack of data on this topic in the south of Brazil, this study aimed to assess the prevalence of Candida spp. in the cervical-vaginal mucosa of patients treated at a university hospital in southern Rio Grande do Sul, as well as the etiology and the susceptibility of the isolates against fluconazole, itraconazole, miconazole and nystatin. Samples were collected at the gynecology clinic of the Federal Hospital of the University of Rio Grande, and the isolates were identified using phenotypic and biochemical tests. The susceptibility analysis was performed according to the CLSI M27-A2 protocol. Of the 263 patients included, Candida spp. was isolated in 27%, corresponding to a prevalence of approximately 15% for both VVC and colonization. More than 60% of the isolates were identified as Candida albicans; C. non-albicans was isolated at a rate of 8.6% in symptomatic patients and 14.3% in asymptomatic patients. The prevalence of resistance against fluconazole and itraconazole was 42% and 48%, respectively; the minimal inhibitory concentration of miconazole ranged from 0.031 to 8 µg/mL, and that of nystatin ranged from 2 to >16 µg/mL. The high rate of resistance to triazoles observed in our study suggests the necessity of the association of laboratory exams to clinical diagnosis to minimize the practice of empirical treatments that can contribute to the development of resistance in the isolates.

  6. Influence of the host contact sequence on the outcome of competition among aspergillus flavus isolates during host tissue invasion.

    Science.gov (United States)

    Mehl, H L; Cotty, P J

    2011-03-01

    Biological control of aflatoxin contamination by Aspergillus flavus is achieved through competitive exclusion of aflatoxin producers by atoxigenic strains. Factors dictating the extent to which competitive displacement occurs during host infection are unknown. The role of initial host contact in competition between pairs of A. flavus isolates coinfecting maize kernels was examined. Isolate success during tissue invasion and reproduction was assessed by quantification of isolate-specific single nucleotide polymorphisms using pyrosequencing. Isolates were inoculated either simultaneously or 1 h apart. Increased success during competition was conferred to the first isolate to contact the host independent of that isolate's innate competitive ability. The first-isolate advantage decreased with the conidial concentration, suggesting capture of limited resources on kernel surfaces contributes to competitive exclusion. Attempts to modify access to putative attachment sites by either coating kernels with dead conidia or washing kernels with solvents did not influence the success of the first isolate, suggesting competition for limited attachment sites on kernel surfaces does not mediate first-isolate advantage. The current study is the first to demonstrate an immediate competitive advantage conferred to A. flavus isolates upon host contact and prior to either germ tube emergence or host colonization. This suggests the timing of host contact is as important to competition during disease cycles as innate competitive ability. Early dispersal to susceptible crop components may allow maintenance within A. flavus populations of genetic types with low competitive ability during host tissue invasion.

  7. Molecular Characterization and Antimicrobial Activity of an Endolichenic Fungus, Aspergillus sp. Isolated from Parmelia caperata of Similipal Biosphere Reserve, India.

    Science.gov (United States)

    Padhi, Srichandan; Das, Devaranjan; Panja, Suraj; Tayung, Kumananda

    2017-06-01

    Endolichenic fungi are microbes that inhabit healthy inner lichen tissues without any disease symptoms. They have been reported to produce new and interesting bioactive metabolites. In the present study, an endolichenic fungus frequently isolated from surface-sterilized lichen thallus of Parmelia caperata has been described. The fungus was identified as Aspergillus tubingensis based on morphological traits and ITS rDNA sequence. Crude metabolites extracted from the culture broth exhibited considerable antimicrobial activity against a panel of clinically significant human pathogens. The fungus showed optimum antimicrobial activity in PDB medium in day 7 of incubation period. PDB medium amended with 1 % NaCl and at alkaline pH was found to be optimal for antimicrobial metabolites production. Enhanced activity was observed when the fungus was exposed briefly to a heat shock of 60 °C during incubation. The metabolites showed optimum λ-max at 214 nm with an absorbance value of 1.589. Molecular characterization of the isolate was carried out by ITS phylogeny and ITS2 secondary structure analyses. The phylogenetic trees based on both ITS rDNA and ITS2 sequences showed the isolate within the clade A. tubingensis. Considering the ubiquity and ambiguity in identifying Aspergillus species of different lifestyles, a method to differentiate pathogenic and endophytic Aspergillus at species level was developed using ITS2 secondary structure analysis. The results showed common folding pattern in the secondary structures with a helix and a 5' dangling end found to be highly conserved. Certain features in the secondary structure like multi-bulges and a symmetric interior loop were observed to be unique which distinguish our isolate from other A. tubingensis.

  8. Isolation, identification and toxigenic potential of ochratoxin A-producing Aspergillus species from coffee beans grown in two regions of Thailand

    DEFF Research Database (Denmark)

    Noonim, P.; Mahakarnchanakul, W.; Nielsen, Kristian Fog

    2008-01-01

    In 2006 and 2007, 32 Thai dried coffee bean samples (Coffea arabica) from two growing sites of Chiang Mai Province, and 32 Thai dried coffee bean samples (Coffea canephora var. robusta) from two growing sites of Chumphon Province, Thailand, were collected and assessed for the distribution of fungi...... with the potential to produce ochratoxin A (OTA). The overall percentage of fungal contamination in coffee was 98% and reduced to 60% after surface disinfection. There were remarkable ecological differences in the composition of ochratoxigenic species present in these two regions. Arabica coffee bean samples from...... the North had an average of 78% incidence of colonization with Aspergillus of section Circumdati with Aspergillus westerdijkiae and A. melleus as the predominant species. Aspergillus spp. of section Nigri were found in 75% of the samples whereas A. ochraceus was not detected. Robusta coffee beans from...

  9. Synergism of Plant Compound With Traditional Antimicrobials Against Streptococcus spp. Isolated From Bovine Mastitis

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    Natasha L. Maia

    2018-06-01

    Full Text Available Mastitis is an inflammation of the mammary gland that causes major losses in the dairy industry. Streptococcus spp. are among the main agents of this disease. Increased resistance to antibiotics is one of the causes of therapeutic failure. Plants, due to their broad chemodiversity, are an interesting source of new molecules with antibacterial activity. Using these compounds along with traditional antibiotics is a possible method for reversing resistance. The objective of this work was to determine the interactions between the activities of guttiferone-A and 7-epiclusianone, two active substances isolated from the fruits of Garcinia brasiliensis, and traditional antibiotics against Streptococcus spp. isolated from bovine mastitis and known to be resistant to them. First, the MIC for the antibiotics and bioactive compounds was determined, followed by their activities, alone and in combination. Then, their cytotoxicity was measured in bovine mammary epithelial cells. Finally, molecular docking simulations were performed to elucidate molecular details of the interactions between β-lactamase and the compounds binding to it (clavulanic acid, ampicillin, 7-epiclusianone, and guttiferone-A. The bacterial isolates were resistant to ampicillin and gentamicin. Both antibiotics showed predominantly synergistic antibacterial activities in combination with guttiferone-A or 7-epiclusianone. These two active substances were not cytotoxic at synergistic concentrations and both showed strong binding to β-lactamase, which may explain the reversal of ampicillin resistance. These substances are promising for the treatment of bovine mastitis.

  10. Virulotyping of Shigella spp. isolated from pediatric patients in Tehran, Iran.

    Science.gov (United States)

    Ranjbar, Reza; Bolandian, Masomeh; Behzadi, Payam

    2017-03-01

    Shigellosis is a considerable infectious disease with high morbidity and mortality among children worldwide. In this survey the prevalence of four important virulence genes including ial, ipaH, set1A, and set1B were investigated among Shigella strains and the related gene profiles identified in the present investigation, stool specimens were collected from children who were referred to two hospitals in Tehran, Iran. The samples were collected during 3 years (2008-2010) from children who were suspected to shigellosis. Shigella spp. were identified throughout microbiological and serological tests and then subjected to PCR for virulotyping. Shigella sonnei was ranking first (65.5%) followed by Shigella flexneri (25.9%), Shigella boydii (6.9%), and Shigella dysenteriae (1.7%). The ial gene was the most frequent virulence gene among isolated bacterial strains and was followed by ipaH, set1B, and set1A. S. flexneri possessed all of the studied virulence genes (ial 65.51%, ipaH 58.62%, set1A 12.07%, and set1B 22.41%). Moreover, the pattern of virulence gene profiles including ial, ial-ipaH, ial-ipaH-set1B, and ial-ipaH-set1B-set1A was identified for isolated Shigella spp. strains. The pattern of virulence genes is changed in isolated strains of Shigella in this study. So, the ial gene is placed first and the ipaH in second.

  11. Local isolate of Saccharomyces cerevisiae as biocompetitive agent of Aspergillus flavus

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    Eni Kusumaningtyas

    2006-12-01

    Full Text Available Aspergillus flavus is a toxigenic fungus that contaminates feed and influences the animal health. Saccharomyces cerevisiae can be used as a biocompetitive agent to control the contamination. The ability of local isolate of S. cerevisiae as a biocompetitive agent for A. flavus was evaluated. A. flavus (30ml was swept on Sabouraud dextrose agar (SDA, while S. cerevisiae was swept on its left and right. Plates were incubated at 28oC for nine days. Lytic activity of S. cerevisiae was detected by pouring its suspension on the centre of the cross streaks of A. flavus. Plates were incubated at 28oC for five days. Growth inhibition of A. flavus by S. cerevisiae was determined by mixing the two fungi on Potato dextrose broth and incubated at 28oC for 24 hours. Total colony of A. flavus were then observed at incubation time of 2, 4, 6 and 24 hours by pour plates method on the SDA plates and incubated on 28oC for two days. Growth of hyphae of A. flavus sweep were inhibited with the swept of S. cerevisiae. The width of A. flavus colony treated with S. cerevisiae is narrower (3,02 cm than that of control ( 4,60 cm. The growth of A. flavus was also inhibited on the centre of cross streak where the S. cerevisiae poured. S. cerevisiae gradually reduced the colony number of A. flavus in the mixed culture of broth fungi ie. 14 x 103 CFU/ml while colony number of control is 80 x 103 CFU/ml. Results showed that S. cerevisiae could be used as biocompetitive agent of A. flavus.

  12. The polyphasic description of a Desmodesmus spp. isolate with the potential of bioactive compounds production

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    El Semary, NA.

    2011-01-01

    Full Text Available A polyphasic approach was applied to describe a colony-forming Desmodesmus species collected from the Nile River, Maadi area, Helwan district, Egypt. The isolate grows best at moderate temperature and relatively high light intensity. The phenotypic features revealed the presence of both unicellular and colonial forms of the isolate and the latter form was either 2-4 celled. Cells were 4-6 mm ± 0.5 at their widest point and 11-15 mm ± 0.48 in their length with spiny projections that encircled the cells. Cells were heavily-granulated and enclosed within common mucilaginous sheath. Colonial forms were developed through production of daughter cells within mother cell. Molecular analysis using 18S rRNA gene showed some similarity to its nearest relative (Desmodesmus communis whereas the phylogenetic analyses clustered it together with other Desmodesmus spp. and away from Scenedesmus spp. from the database. However, the use of ITS-2 as a phylotaxonomic marker proved to be more resolving and confirmed the generic identity of the isolate as Desmodesmus spp. The fatty acid composition revealed the presence of saturated palmitic fatty acid as the most abundant component followed by monounsaturated palmitoleic acid whereas the polyunsaturated fatty acids were in relatively low abundance. The palmitoleic acid in particular is suggested to be involved in active defense mechanism. The phytochemical screening revealed the presence of alkaloids and saponins and absence of tannins. Fractions of methanolic extracts showed antimicrobial activities against pathogenic bacterial strains including multi-drug resistant ones. This study documents the presence of this strain in the River Nile and highlights its biotechnological potential as a source of bioactive compounds.

  13. Tetracycline and Azithromycin Resistance Investigation on Shigella spp. Isolated from the Stool of Children with Diarrhea in Tehran, Iran

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    Shadi Shahsavan

    2016-10-01

    Full Text Available Background & objectives: Shigella spp. are gram negative bacteria that can cause shigellosis in human. It is important in young children as well as elderly and immunocompromised people. Threatening complications can occur in severe cases with multidrug resistance species. It has been observed that Shigella spp. have become resistant to antibiotics like other bacteria. Investigation of resistance to azithromycin, tetracycline and pattern of resistance are the objectives of this study. Methods: Fifty isolates of Shigella spp. which have been collected from three hospitals in Tehran were studied. Isolates identified and confirmed as Shigella spp. by biochemical, serological and molecular methods (ipaH, wbgz, rfc genes. Antimicrobial susceptibility test was performed for ampicillin, azithromycin, ciprofloxacin, doxycycline, levofloxacin, minocycline, nalidixic acid, norfloxacin, streptomycin, trimethoprim-sulfamethoxazole and tetracycline by disc agar diffusion method. Minimal inhibition concentrations were performed for azithromycin and tetracycline. Results: From a total of 50 Shigella spp. isolates, 16% of them were Shigella flexneri and 84% Shigella sonnei. The majority of isolates were multidrug resistant. The most resistance was seen to doxycycline, streptomycin, trimethoprim-sulfamethoxazole and tetracycline. Resistance to azithromycin was 6%  and all of the isolates were susceptible to norfloxacin and levofloxacin. Nine patterns of resistance were revealed to these isolates. Conclusion: High resistance to tetracycline was observed and resistance to azithromycin as an alternative treatment choice was also considerable.

  14. Phenotypic and molecular characterization of antimicrobial resistance in Enterobacter spp. isolates from companion animals in Japan.

    Science.gov (United States)

    Harada, Kazuki; Shimizu, Takae; Mukai, Yujiro; Kuwajima, Ken; Sato, Tomomi; Kajino, Akari; Usui, Masaru; Tamura, Yutaka; Kimura, Yui; Miyamoto, Tadashi; Tsuyuki, Yuzo; Ohki, Asami; Kataoka, Yasushi

    2017-01-01

    The emergence of antimicrobial resistance among Enterobacter spp., including resistance to extended-spectrum cephalosporins (ESC), is of great concern in both human and veterinary medicine. In this study, we investigated the prevalence of antimicrobial resistance among 60 isolates of Enterobacter spp., including E. cloacae (n = 44), E. aerogenes (n = 10), and E. asburiae (n = 6), from clinical specimens of dogs and cats from 15 prefectures in Japan. Furthermore, we characterized the resistance mechanisms harbored by these isolates, including extended-spectrum β-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR); and assessed the genetic relatedness of ESC-resistant Enterobacter spp. strains by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Antimicrobial susceptibility testing demonstrated the resistance rates to ampicillin (93.3%), amoxicillin-clavulanic acid (93.3%), cefmetazole (93.3%), chloramphenicol (46.7%), ciprofloxacin (43.3%), tetracycline (40.0%), ceftazidime (33.3%), cefotaxime (33.3%), trimethoprim/sulfamethoxazole (28.3%), gentamicin (23.3%), and meropenem (0%). Phenotypic testing detected ESBLs in 16 of 18 ESC-resistant E. cloacae isolates but not in the other species. The most frequent ESBL was CTX-M-15 (n = 8), followed by SHV-12 (n = 7), and CTX-M-3 (n = 1). As for AmpC β-lactamases, CMY-2 (n = 2) and DHA-1 (n = 2) were identified in ESC-resistant E. cloacae strains with or without ESBLs. All of the ESC-resistant E. cloacae strains also harbored one or two PMQRs, including qnrB (n = 15), aac(6')-Ib-cr (n = 8), and qnrS (n = 2). Based on MLST and PFGE analysis, E. cloacae clones of ST591-SHV-12, ST171-CTX-M-15, and ST121-CTX-M-15 were detected in one or several hospitals. These results suggested intra- and inter-hospital dissemination of E. cloacae clones co-harboring ESBLs and PMQRs among companion animals. This is the first report on the large-scale monitoring of antimicrobial-resistant isolates

  15. Antimicrobial resistance of Pseudomonas spp. isolated from wastewater and wastewater-impacted marine coastal zone.

    Science.gov (United States)

    Luczkiewicz, Aneta; Kotlarska, Ewa; Artichowicz, Wojciech; Tarasewicz, Katarzyna; Fudala-Ksiazek, Sylwia

    2015-12-01

    In this study, species distribution and antimicrobial susceptibility of cultivated Pseudomonas spp. were studied in influent (INF), effluent (EFF), and marine outfall (MOut) of wastewater treatment plant (WWTP). The susceptibility was tested against 8 antimicrobial classes, active against Pseudomonas spp.: aminoglycosides, carbapenems, broad-spectrum cephalosporins from the 3rd and 4th generation, extended-spectrum penicillins, as well as their combination with the β-lactamase inhibitors, monobactams, fluoroquinolones, and polymyxins. Among identified species, resistance to all antimicrobials but colistin was shown by Pseudomonas putida, the predominant species in all sampling points. In other species, resistance was observed mainly against ceftazidime, ticarcillin, ticarcillin-clavulanate, and aztreonam, although some isolates of Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas pseudoalcaligenes, and Pseudomonas protegens showed multidrug-resistance (MDR) phenotype. Among P. putida, resistance to β-lactams and to fluoroquinolones as well as multidrug resistance become more prevalent after wastewater treatment, but the resistance rate decreased in marine water samples. Obtained data, however, suggests that Pseudomonas spp. are equipped or are able to acquire a wide range of antibiotic resistance mechanisms, and thus should be monitored as possible source of resistance genes.

  16. In vitro effects of various xenobiotics on Fusarium spp. strains isolated from cereals.

    Science.gov (United States)

    Wolny-Koładka, Katarzyna A

    2014-01-01

    This study aimed to determine the susceptibility of Fusarium spp. strains isolated from cereals to selected heavy metals, fungicides and silver nanoparticles. The experiments were conducted using 50 Fusarium strains belonging to five species: F. graminearum, F. culmorum, F. oxysporum, F. sporotrichioides and F. avenaceum. The strains were found to be highly resistant to Pb(2+) and Zn(2+). Medium resistance to Cu(2+) and Mn(2+) and low resistance to Cd(2+) and Fe(3+) was also observed. Among the tested fungicides, formulations containing azoxystrobin, prochloraz and tebuconazole proved to be the most effective in inhibiting the growth of fungi, as they affected fungal growth in each of the applied doses. Susceptibility of Fusarium spp. to nanosilver, demonstrated in this study, shows the legitimacy of using nanostructures as fungicidal agents. The results confirm high diversity of the analyzed fungal species in terms of susceptibility to the tested substances, and encourage to continue research on the resistance of Fusarium spp. to various fungicidal agents.

  17. PREVALENCE AND IDENTIFICATION OF VIBRIO SPP. ISOLATED ON AQUACULTURED GILTHEAD SEA BREAM

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    C. Scarano

    2011-01-01

    Full Text Available The aim of the study was to investigate the prevalence of Vibrio spp isolated from gilthead sea bream (Sparus aurata farmed on sea cages and to identify and characterize the pathogen by molecular techniques. Eighty fish were collected from two hatcheries located on the North-Est Sardinian Mediterranean coast, and microbiological analysis were performed on different body parts such as skin, gills, muscle and intestinal tract. Subsequently 100 pure colonies with typical morphology and phenotypic characteristics were selected and submitted to the molecular identification. The analysis on the prevalence of Vibrio spp showed the effect of the hatchery rearing system (P<0.001, of the date of sampling (P<0.001, and of the body part (P<0.001. All the strains selected were confirmed to be members of the genus Vibrio spp by the molecular method/techinique/identification, whereas the rpoA gene sequence analyses allowed to identify 89 strains belonging to the species Vibrio harveyi, 6 to V. diabolicus, 2 to V. parahaemolyticus and 1 to V. mediterranei.

  18. A Simple Procedure to Evaluate Competitiveness of Toxigenic and Atoxigenic Isolates of Aspergillus flavus in Solid and Liquid Media

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    Mohammad Moradi

    2017-04-01

    Full Text Available Application of atoxigenic strains of Aspergillus flavus to reduce aflatoxin levels is the most successful strategy applied in some agricultural crops. The role of ammonium hydroxide for preliminary screening of the competitiveness of atoxigenic A. flavus isolates to interfere with aflatoxin production by highly toxigenic isolates were evaluated. Out of 270 A. flavus isolates, 17 were detected as true atoxigenic using cultural methods and confirmed by analytical assays from different pistachio agro-ecological zones during 2013. For assessment competitive ability among atoxigenic isolates of A. flavus with highly toxigenic one, rice flour, coconut agar and coconut broth medium substrates were inoculated with mixtures including combinations of toxigenic and atoxigenic isolates, simultaneously. The rice flour substrate was used to quantify the content of aflatoxin in either co-inoculations or toxigenic isolate alone on thin layer chromatography plates with a scanning densitometer. While the culture media were used to determine the intensity of color change on exposing to ammonium hydroxide vapor. The reduction rates of aflatoxin B1 in co-inoculations were varied and ranged from 2%-82%. Based on the intensity of colony color changes, the competitiveness of the isolates was classified into five groups. Atoxigenic isolates with high competitiveness have shown low color changes in culture media and high aflatoxin reduction in TLC assays with a ratio of higher than 78%. The method will facilitate preliminary screening of efficient atoxigenic isolates for mitigation of aflatoxins in food and feed as a cheap, simple and quick method.

  19. Molecular Variability Among Isolates of Prunus Necrotic Ringspot Virus from Different Prunus spp.

    Science.gov (United States)

    Aparicio, F; Myrta, A; Di Terlizzi, B; Pallás, V

    1999-11-01

    ABSTRACT Viral sequences amplified by polymerase chain reaction from 25 isolates of Prunus necrotic ringspot virus (PNRSV), varying in the symptomatology they cause in six different Prunus spp., were analyzed for restriction fragment polymorphisms. Most of the isolates could be discriminated by using a combination of three different restriction enzymes. The nucleotide sequences of the RNA 4 of 15 of these isolates were determined. Sequence comparisons and phylogenetic analyses of the RNA 4 and coat proteins (CPs) revealed that all of the isolates clustered into three different groups, represented by three previously sequenced PNRSV isolates: PV32, PE5, and PV96. The PE5-type group was characterized by a 5' untranslated region that was clearly different from that of the other two groups. The PV32-type group was characterized by an extra hexanucleotide consisting of a duplication of the six immediately preceding nucleotides. Although most of the variability was observed in the first third of the CP, the amino acid residues in this region, which were previously thought to be functionally important in the replication cycle of the virus, were strictly conserved. No clear correlation with the type of symptom or host specificity could be observed. The validity of this grouping was confirmed when other isolates recently characterized by other authors were included in these analyses.

  20. Virulence assessment of Portuguese isolates of potato cyst nematodes (Globodera spp.

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    Maria José M. DA CUNHA

    2012-05-01

    Full Text Available Identification of species and virulence groups of potato cyst nematodes (PCN, Globodera pallida and G. rostochiensis, present in field populations is important in the control of these nematodes by means of resistant cultivars. In order to characterize the virulence of Globodera spp. isolates from Portugal, 43 G. rostochiensis and three G. pallida isolates were evaluated by measuring their multiplication rates on a susceptible potato cultivar and five differential potato genotypes in a growth chamber pot experiment. Principal Component Analysis and Hierarchical Cluster Analysis showed that the reproduction rates were different in terms of both the numbers of eggs and the numbers of cysts produced. Portuguese isolates of PCN were more virulent on genotypes derived from Solanum vernei than on genotypes derived from other Solanum resistance sources, and there was a significant nematode isolate × host genotype interaction. The virulence bioassay clearly distinguished the two PCN species but failed to differentiate isolates into pathotypes. There was a wide and continuous range of virulence to the resistant genotypes, especially in G. rostochiensis isolates.

  1. Isolation of saprophytic Leptospira spp. from a selected environmental water source of Argentina.

    Science.gov (United States)

    Scialfa, Exequiel; Grune, Sylvia; Brihuega, Bibiana; Aguirre, Pablo; Rivero, Marina

    2017-11-29

    Ten Leptospira spp. strains were isolated from water samples from Nievas stream, Olavarría, Buenos Aires province (Argentina). The isolates showed the typical motility and morphology of the genus Leptospira under dark field microscopy, developing in liquid EMJH medium after eight days of incubation at 13°C and 30°C. All isolates were negative by the Multiple Locus Variable Number Tandem Repeat Analysis (MLVA). Molecular identification by 16S rRNA gene sequencing identified all isolates as nonpathogenic leptospires. Four isolates showed a genetic profile identical to that of the reference strain Leptospira biflexa serovar Patoc, and six isolates revealed sequence similarities within the 97-98% range, closely related to Leptospira yanagawae and Leptospira meyeri, respectively. Strains ScialfaASA42, ScialfaASA45, ScialfaASA44, ScialfaASA47, ScialfaASA49, ScialfaASA50 and ScialfaASA51 possibly represent a novel species of the genus Leptospira. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. Isolation and Identification of Pyrene Mineralizing Mycobacterium spp. from Contaminated and Uncontaminated Sources

    International Nuclear Information System (INIS)

    Lease, C.W.M; Bentham, R.H; Gaskin, S.E; Juhasz, A.L

    2011-01-01

    Mycobacterium isolates obtained from PAH-contaminated and uncontaminated matrices were evaluated for their ability to degrade three-, four- and five-ring PAHs. PAH enrichment studies were prepared using pyrene and inocula obtained from manufacturing gas plant (MGP) soil, uncontaminated agricultural soil, and faeces from Macropus fuliginosus (Western Grey Kangaroo). Three pyrene-degrading microorganisms isolated from the corresponding enrichment cultures had broad substrate ranges, however, isolates could be differentiated based on surfactant, phenol, hydrocarbon and PAH utilisation. 16S rRNA analysis identified all three isolates as Mycobacterium sp. The Mycobacterium spp. could rapidly degrade phenanthrene and pyrene, however, no strain had the capacity to utilise fluorene or benzo[a]pyrene. When pyrene mineralisation experiments were performed, 70-79% of added 14 C was evolved as 14 CO 2 after 10 days. The present study demonstrates that PAH degrading microorganisms may be isolated from a diverse range of environmental matrices. The present study demonstrates that prior exposure to PAHs was not a prerequisite for PAH catabolic activity for two of these Mycobacterium isolates.

  3. Mycotoxin production and predictive modelling kinetics on the growth of Aspergillus flavus and Aspergillus parasiticus isolates in whole black peppercorns (Piper nigrum L).

    Science.gov (United States)

    Yogendrarajah, Pratheeba; Vermeulen, An; Jacxsens, Liesbeth; Mavromichali, Evangelia; De Saeger, Sarah; De Meulenaer, Bruno; Devlieghere, Frank

    2016-07-02

    The growth and mycotoxin production of three Aspergillus flavus isolates and an Aspergillus parasiticus isolate were studied in whole black peppercorns (Piper nigrum L.) using a full factorial design with seven water activity (aw) (0.826-0.984) levels and three temperatures (22, 30 and 37°C). Growth rates and lag phases were estimated using linear regression. Diverse secondary models were assessed for their ability to describe the radial growth rate as a function of individual and combined effect of aw and temperature. Optimum radial growth rate ranged from 0.75±0.04 to 2.65±0.02mm/day for A. flavus and 1.77±0.10 to 2.50±0.10mm/day for A. parasiticus based on the Rosso cardinal estimations. Despite the growth failure of some isolates at marginal conditions, all the studied models showed good performance to predict the growth rates. Validation of the models was performed on independently derived data. The bias factors (0.73-1.03), accuracy factors (0.97-1.36) and root mean square error (0.050-0.278) show that the examined models are conservative predictors of the colony growth rate of both fungal species in black peppers. The Rosso cardinal model can be recommended to describe the individual aw effect while the extended Gibson model was the best model for describing the combined effect of aw and temperature on the growth rate of both fungal species in peppercorns. Temperature optimum ranged from 30 to 33°C, while aw optimum was 0.87-0.92 as estimated by multi-factorial cardinal model for both species. The estimated minimum temperature and aw for A. flavus and A. parasiticus for growth were 11-16°C and 0.73-0.76, respectively, hence, achieving these conditions should be considered during storage to prevent the growth of these mycotoxigenic fungal species in black peppercorns. Following the growth study, production of mycotoxins (aflatoxins B1, B2, G1, G2, sterigmatocystin and O-methyl sterigmatocystin (OMST)) was quantified using LC-MS/MS. Very small

  4. Isolation of Notoamide S and Enantiomeric 6-epi-Stephacidin A from the Fungus Aspergillus amoenus: Biogenetic Implications

    DEFF Research Database (Denmark)

    Kato, Hikaru; Nakahara, Takashi; Sugimoto, Kayo

    2015-01-01

    Notoamide S has been hypothesized to be a key biosynthetic intermediate for characteristic metabolites stephacidin A, notoamide B, and versicolamide B in Aspergillus sp. but has not yet been isolated. The isolation of notoamide S and an enantiomeric mixture of 6-epi-stephacidin A enriched with th...

  5. Molecular Characterization and Phylogenetic Analysis of Anaplasma spp. and Ehrlichia spp. Isolated from Various Ticks in Southeastern and Northwestern Regions of Iran.

    Science.gov (United States)

    Jafar Bekloo, Ahmad; Ramzgouyan, Maryam Roya; Shirian, Sadegh; Faghihi, Faezeh; Bakhshi, Hassan; Naseri, Fatemeh; Sedaghat, Mehdi; Telmadarraiy, Zakkyeh

    2018-05-01

    Anaplasma/Ehrlichia species are tick-transmitted pathogens that cause infections in humans and numerous domestic and wild animal species. There is no information available on the molecular characteristics and phylogenetic position of Anaplasma/Ehrlichia spp. isolated from tick species from different geographic locations in Iran. The aim of this study was to determine the prevalence, molecular characteristics, and phylogenetic relationship of both Anaplasma spp. and Ehrlichia spp. in tick species isolated from different domestic animals from two different geographical locations of Iran. A total of 930 ticks were collected from 93 cattle, 250 sheep, and 587 goats inhabiting the study areas. The collected ticks were then investigated for the presence of Anaplasma/Ehrlichia spp. using nested PCR based on the 16S rRNA gene, followed by sequencing. Sequence analysis was done based on the data published in the GenBank on Anaplasma/Ehrlichia spp. isolates using bioinformatic tools such as the standard nucleotide BLAST. Genome of Anaplasma or Ehrlichia spp. was detected in 14 ticks collected in Heris, including 5 Dermacentor marginatus, 1 Haemaphysalis erinacei, 3 Hyalomma anatolicum, and 4 Rhipicephalus sanguineus, also in 29 ticks collected in Chabahar, including 14 R. sanguineus, 8 D. marginatus, 3 Hyalomma Anatolicum, and 4 Hyalomma dromedarii. Partial analysis of the 16S rRNA gene sequence of positive samples collected from goats and sheep showed that they were infected with Anaplasma/Ehrlichia spp. that were 94-98% identical to ovine Anaplasma and 91-96% identical to Neoehrlichia and Ehrlichia spp. The various ticks identified in this study suggest the possible emergence of tick-borne diseases in animals and humans in these regions. R. sanguineus and D. marginatus seem to be predominant vectors responsible for anaplasmosis in these regions. Partial sequence analysis of the 16S rRNA gene showed that A. ovis is genetically polymorphic in these regions. Furthermore, an

  6. Characterization and biological treatment of colored textile wastewaters from the typical Tunisian hat Chechia dyeing using newly isolated Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Hajer Barouni

    2016-09-01

    Full Text Available This study aimed to characterize and investigate, for the first time, the treatment of real colored wastewaters from the artisanal dyeing of the typical Tunisian hat Chechia, using a newly isolated fungal strain. This textile effluent was a mixture called Mix of colored wastewaters from the three main types of Chechia. The major pollutant of the Mix was the toxic Azo dye Amaranth Acid or Acid Red 27. The fungal strain that made the cleanup was discovered in a Chechia dyeing wastewater’s container and identified by ITS rDNA gene sequencing. This isolated Aspergillus niger showed interesting performances on the demonstration of Chechia wastewater’s biodegradation in batch cultures. In order to understand the effect of agitation, Mix dilution and inoculum size on decolourisation and pollution removal, a full factorial experimental design 23 was set up. At the optimal conditions which were 20% inoculum size, 25% Chechia Mix dilution and an agitation of 100 rpm, Aspergillus niger was able to remove color as high as 70.18±2.84% at an initial dye concentration of 1346.6±0.01 mg/L, and to reduce COD to 74.17±14.52% at an initial COD of 4157±422 mg/L. FT-IR spectra analysis confirmed the decolourisation by biodegradation and transformation of the dyes. The treatment by the isolated Aspergillus niger could be successfully applied as a sustainable method to solve one of handicraft dyeing plants environmental management issues.

  7. Heavy metal and antibiotic resistance of Acinetobacter spp. isolated from diesel fuel polluted

    Directory of Open Access Journals (Sweden)

    Kais Kassim Ghaima

    2018-04-01

    Full Text Available Heavy metals pollution of soil and wastewater is a global problem that threatens the environment as they are not degraded or removed and the potential threat to human health comes from the multiple resistances to heavy metals and antibiotics among bacterial populations. The present study was aimed to isolate and identify multiple metal/antibiotic resistant Acinetobacter spp. from diesel fuel polluted soil of Al-Dora, Baghdad, Iraq. Initially, a total of 24 bacterial cultures (coded KNZ–1 to KNZ–24 were isolated and identified up to genus level as Acinetobacter by morphological, physiological and biochemical characteristics. Screening of heavy metals resistant Acinetobacter were conducted by streaking the isolates on nutrient agar plates supplemented with different concentrations: 10, 25, 50 and 100mg/L of the three heavy metals; Hg2+, Cd2+ and Pb2+. Out of 24 isolates, 6 (25% isolates (KNZ–3, KNZ–5, KNZ–8, KNZ–12, KNZ–16 and KNZ–21 were selected as a multiple heavy metal resistant (MHMR Acinetobacter with maximum tolerable concentrations (MTCs; 100–200mg/L for Hg2+, 300-600mg/L for Cd2+ and 100–300mg/L for Pb2+. Antibiotic resistance pattern of the selected MHMR isolates was determined by Kirby-Bauer disc diffusion method against 12 different antibiotics belonging to 7 classes. Out of 6 isolates, 4 isolates were multidrug resistance (MDR with varying degrees. Among them isolate, KNZ–16 showed a wide range of resistance to all tested antibiotics except Levofloxacin and Imipenem. It was concluded that dual resistant Acinetobacter is useful in the bioremediation of environments polluted with heavy metals especially the biodegradation of organic pollutants.

  8. Cr(VI) reduction from contaminated soils by Aspergillus sp. N2 and Penicillium sp. N3 isolated from chromium deposits.

    Science.gov (United States)

    Fukuda, Tsubasa; Ishino, Yasuhiro; Ogawa, Akane; Tsutsumi, Kadzuyo; Morita, Hiroshi

    2008-10-01

    Aspergillus sp. N2 and Penicillium sp. N3 are chromate-resistant filamentous fungi that were isolated from Cr(VI) contaminated soil based on their ability to decrease hexavalent chromium levels in the growth medium. After 120 h of growth in a medium containing 50 ppm Cr(VI) at near neutral pH, Aspergillus sp. N2 reduced the Cr(VI) concentration by about 75%. Penicillium sp. N3 was able to reduce the Cr(VI) concentration by only 35%. However, Penicillium sp. N3 reduced the Cr(VI) concentration in the medium by 93% under acidic conditions. Interestingly, the presence of Cu(II) enhanced the Cr(VI) reducing ability of Aspergillus sp. N2 and Penicillium sp. N3 at near neutral pH. Aspergillus sp. N2 and Penicillium sp. N3 reduced the Cr(VI) concentration in the growth medium to a virtually undetectable level within 120 h. For both Aspergillus sp. N2 and Penicillium sp. N3, mycelial seed cultures were more efficient at Cr(VI) reduction than conidium seed cultures. The mechanisms of Cr(VI) reduction in Aspergillus sp. N2 and Penicillium sp. N3 were enzymatic reduction and sorption to mycelia. Enzymatic activity contributed significantly to Cr(VI) reduction. Aspergillus sp. N2 and Penicillium sp. N3 reduced the levels of Cr(VI) in polluted soil samples, suggesting that these strains might be useful for cleaning up chromium-contaminated sites.

  9. The Use of Trichoderma spp. for Controlling the Growth and Aflatoxin Production of Aspergillus parasiticus in Agricultural Products

    International Nuclear Information System (INIS)

    Thanaboripat, Dusanee; Piadiang, Nattaya; Qian, Yang

    2006-09-01

    Various strains of Trichoderma spp. were screened from soils and irradiated by gamma ray. After the irradiation all strains were tested for an ability to inhibit the growth of aflatoxin producing fungi (A. parasiticus IMI 105266 and A. flavus IMI 24268). The results indicate that Trichoderma virde S84-1 480526 I08(1) was the most effective strain in controlling the growth of these two fungi on PDA.

  10. Comparative analysis on antibiotic resistance characteristics of Listeria spp. and Enterococcus spp. isolated from laying hens and eggs in conventional and organic keeping systems in Bavaria, Germany.

    Science.gov (United States)

    Schwaiger, K; Schmied, E-M V; Bauer, J

    2010-05-01

    By investigating the prevalence and antimicrobial resistance characteristics of Gram-positive bacteria from organic and conventional keeping systems of laying hens, it was to be determined to what extent these properties are influenced by the different systems. For this purpose, a total of 799 cloacal swabs and 800 egg samples were examined. Prevalences for all selected bacteria from cloacal swabs were much the same for both organic and caged birds: Listeria spp.1.3%[org] versus 1.6%[con]; Enterococcus spp. 95.5%[org] versus 97.5%[con]. Egg contents and eggshells were generally contaminated to a lesser extent, primarily with Enterococcus spp. Listeria isolates were susceptible to almost all tested antibiotics, only three Listeria innocua from conventional keepings were resistant to clindamycin; one isolate additionally to imipenem. High percentages of Enterococcus faecalis were resistant to doxycycline and macrolides. Enterococcus faecium proved to have high resistance rates to clindamycin, fosfomycin and erythromycin; 9.1% were even resistant to the reserve antibiotic synercid. Further, Enterococcus spp. showed higher resistance rates to doxycycline, erythromycin, fosfomycin and rifampicin. No glycopeptide resistant enterococci were detected. A correlation between keeping system and resistance/susceptibility rates could be demonstrated. In detail, E. faecalis from organic laying hen husbandries showed significant lower resistance prevalences to tylosin, streptomycin and doxycycline; susceptibility rates were higher for enrofloxacin and ciprofloxacin. Rifampicin and imipenem were more effective in isolates from conventional keepings (P < 0.05). The amounts of resistant isolates of the Enterococcus raffinosus from organic farms were significantly lower, the amounts of sensitive isolates were significantly higher than from conventional farms concerning eight antibiotics (P < 0.05). When comparing the susceptibility/resistance rates, as well as the mean minimum

  11. Lethality of cytochalasin B and other compounds isolated from fungus Aspergillus sp. (Trichocomaceae) endophyte of Bauhinia guianensis (Fabaceae).

    Science.gov (United States)

    Feitosa, André de O; Dias, Amanda Cristina S; Ramos, Gisele da C; Bitencourt, Heriberto R; Siqueira, José Edson S; Marinho, Patrícia Santana B; Barison, Andersson; Ocampos, Fernanda M M; Marinho, Andrey Moacir do R

    Endophytic fungi are fungi that colonize internal tissues of plants; several biologically active compounds have been isolated from these fungi. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, this study aimed the isolation and structural identification of ergosterol (1), ergosterol peroxide (2), mevalonolactone (3), cytochalasin B (4) and cytochalasin H (5) from Aspergillus sp. EJC 04, an endophytic fungus from Bauhinia guianensis. The cytochalasin B (4) and the diacetate derivative of cytochalasin B (4a) showed high lethality in the brine shrimp assay. This is the first occurrence of cytochalasins in Amazonian endophytic fungi from B. guianensis. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  12. Isolation and identification of Salmonella spp. in environmental water by molecular technology in Taiwan

    Science.gov (United States)

    Kuo, Chun Wei; Hao Huang, Kuan; Hsu, Bing Mu; Tsai, Hsien Lung; Tseng, Shao Feng; Shen, Tsung Yu; Kao, Po Min; Shen, Shu Min; Chen, Jung Sheng

    2013-04-01

    Salmonella spp. is one of the most important causal agents of waterborne diseases. The taxonomy of Salmonella is very complicated and its genus comprises more than 2,500 serotypes. The detection of Salmonella in environmental water samples by routines culture methods using selective media and characterization of suspicious colonies based on biochemical tests and serological assay are generally time consuming. To overcome this drawback, it is desirable to use effective method which provides a higher discrimination and more rapid identification about Salmonella in environmental water. The aim of this study is to investigate the occurrence of Salmonella using molecular technology and to identify the serovars of Salmonella isolates from 70 environmental water samples in Taiwan. The analytical procedures include membrane filtration, non-selective pre-enrichment, selective enrichment of Salmonella. After that, we isolated Salmonella strains by selective culture plates. Both selective enrichment and culture plates were detected by Polymerase Chain Reaction (PCR). Finally, the serovars of Salmonella were confirmed by using biochemical tests and serological assay. In this study, 15 water samples (21.4%) were identified as Salmonella by PCR. The positive water samples will further identify their serotypes by culture method. The presence of Salmonella in environmental water indicates the possibility of waterborne transmission in drinking watershed. Consequently, the authorities need to provide sufficient source protection and to maintain the system for disease prevention. Keywords: Salmonella spp., serological assay, PCR

  13. Identification of Staphylococcus spp. isolated during the ripening process of a traditional minas cheese

    Directory of Open Access Journals (Sweden)

    B.M. Borelli

    2011-04-01

    Full Text Available The population dynamics of Staphylococcus spp. was studied during the ripening of Canastra Minas cheese at three farms located in the State of Minas Gerais, Brazil. The presence of coagulase (coa, thermonuclease (nuc, and enterotoxin (sea, seb, sec, and sed genes was investigated in Staphylococcus strains isolated during the 60-day cheese-ripening period. The presence of the staphylococcal enterotoxins A, C, and D was also investigated in the cheese samples. Cheese samples that were matured for 0, 7, 15, 30, and 45 days presented staphylococci counts from 10³ to 10(8cfu/g. All isolates considered coagulase-positive by physiological tests had the coa gene. However, no association was observed between the results obtained with biochemical tests and those obtained by PCR using gene-specific primers for coagulase-negative strains. Coagulase and thermonuclease genes occurred simultaneously in 41.3% of Staphylococcus spp. tested. None of the investigated Staphylococcus strains expressed enterotoxins SEA, SEB, SEC, and SED. Enterotoxins A, C, and D were not detected in any of the cheese samples.

  14. Identification and susceptibility of clinical isolates of Candida spp. to killer toxins

    Directory of Open Access Journals (Sweden)

    E. Robledo-Leal

    2018-02-01

    Full Text Available Abstract Although invasive infections and mortality caused by Candida species are increasing among compromised patients, resistance to common antifungal agents is also an increasing problem. We analyzed 60 yeasts isolated from patients with invasive candidiasis using a PCR/RFLP strategy based on the internal transcribed spacer (ITS2 region to identify different Candida pathogenic species. PCR analysis was performed from genomic DNA with a primer pair of the ITS2-5.8S rDNA region. PCR-positive samples were characterized by RFLP. Restriction resulted in 23 isolates identified as C. albicans using AlwI, 24 isolates as C. parapsilosis using RsaI, and 13 as C. tropicalis using XmaI. Then, a group of all isolates were evaluated for their susceptibility to a panel of previously described killer yeasts, resulting in 75% being susceptible to at least one killer yeast while the remaining were not inhibited by any strain. C. albicans was the most susceptible group while C. tropicalis had the fewest inhibitions. No species-specific pattern of inhibition was obtained with this panel of killer yeasts. Metschnikowia pulcherrima, Pichia kluyveri and Wickerhamomyces anomalus were the strains that inhibited the most isolates of Candida spp.

  15. Genotypic and phenotypic diversity of Bacillus spp. isolated from steel plant waste

    Directory of Open Access Journals (Sweden)

    Chartone-Souza Edmar

    2008-10-01

    Full Text Available Abstract Background Molecular studies of Bacillus diversity in various environments have been reported. However, there have been few investigations concerning Bacillus in steel plant environments. In this study, genotypic and phenotypic diversity and phylogenetic relationships among 40 bacterial isolates recovered from steel plant waste were investigated using classical and molecular methods. Results 16S rDNA partial sequencing assigned all the isolates to the Bacillus genus, with close genetic relatedness to the Bacillus subtilis and Bacillus cereus groups, and to the species Bacillus sphaericus. tDNA-intergenic spacer length polymorphisms and the 16S–23S intergenic transcribed spacer region failed to identify the isolates at the species level. Genomic diversity was investigated by molecular typing with rep (repetitive sequence based PCR using the primer sets ERIC2 (enterobacterial repetitive intergenic consensus, (GTG5, and BOXAIR. Genotypic fingerprinting of the isolates reflected high intraspecies and interspecies diversity. Clustering of the isolates using ERIC-PCR fingerprinting was similar to that obtained from the 16S rRNA gene phylogenetic tree, indicating the potential of the former technique as a simple and useful tool for examining relationships among unknown Bacillus spp. Physiological, biochemical and heavy metal susceptibility profiles also indicated considerable phenotypic diversity. Among the heavy metal compounds tested Zn, Pb and Cu were least toxic to the bacterial isolates, whereas Ag inhibited all isolates at 0.001 mM. Conclusion Isolates with identical 16S rRNA gene sequences had different genomic fingerprints and differed considerably in their physiological capabilities, so the high levels of phenotypic diversity found in this study are likely to have ecological relevance.

  16. Reduction of Aspergillus spp. and aflatoxins in peanut sauce processing by oil-less frying of chilli powder and retort processing.

    Science.gov (United States)

    Farawahida, A H; Jinap, S; Nor-Khaizura, M A R; Samsudin, N I P

    2017-12-01

    Among the many roles played by small and medium enterprises (SMEs) in the food industry is the production of heritage foods such as peanut sauce. Unfortunately, the safety of peanut sauce is not always assured as the processing line is not controlled. Peanut sauce is usually made of peanuts and chilli, and these commodities are normally contaminated with Aspergillus spp. and aflatoxins (AFs). Hence, the objective of this study was to evaluate the practices related to reduction of AF hazard and the effect of interventions in peanut sauce processing. Peanut samples were collected from each step of peanut sauce processing from a small peanut sauce company according to four designs: (1) control; (2) oil-less frying of chilli powder; (3) addition of retort processing; and (4) combination of oil-less frying of chilli powder and retort processing. Oil-less frying of chilli powder (Design 2) reduced total AFs by 33-41%, retort processing (Design 3) reduced total AFs by 49%, while combination of these two thermal processes (Design 4) significantly reduced total AFs, by 57%. The present work demonstrated that Design 4 yielded the highest reduction of total AFs and is therefore recommended to be employed by SME companies.

  17. Antimicrobial resistance of fecal Salmonella spp. isolated from all phases of pig production in 20 herds in Alberta and Saskatchewan

    OpenAIRE

    Rosengren, Leigh B.; Waldner, Cheryl L.; Reid‐Smith, Richard J.; Checkley, Sylvia L.; McFall, Margaret E.; Rajíc, Andrijana

    2008-01-01

    Salmonella spp. (n = 468), isolated from the feces of sows, nursery, and grow‐finish pigs in 20 farrow‐to‐finish herds in Alberta and Saskatchewan, were tested for susceptibility to 16 antimicrobials. No resistance was identified to amikacin, amoxicillin‐clavulanic acid, ceftiofur, ceftriaxone, ciprofloxacin or nalidixic acid, and less than 1% of the isolates were resistant to cefoxitin and gentamicin. Isolates were most commonly resistant to tetracycline (35%) and sulfamethoxazole (27%). Ove...

  18. Biodegradation of imidacloprid in liquid media by an isolated wastewater fungus Aspergillus terreus YESM3.

    Science.gov (United States)

    Mohammed, Youssef M M; Badawy, Mohammed E I

    2017-10-03

    In the present study, a new fungal strain capable of imidacloprid degradation was isolated from agricultural wastewater drain. The fungal strain of YESM3 was identified as Aspergillus terreus based on ITS1-5.8S rDNA-ITS2 gene sequence by PCR amplification of a 500 bp sequence. Screening of A. terreus YESM3 to the insecticide imidacloprid tolerance was achieved by growing fungus in Czapek Dox agar for 6 days at 28°C. High values (1.13 and 0.94 cm cm -1 ) of tolerance index (TI) were recorded at 25 and 50 mg L -1 of imidacloprid, respectively in the presence and absence of sucrose. However, at 400 mg L -1 the fungus did not grow. Effects of the imidacloprid concentration, pH, and inoculum size on the biodegradation percentage were tested using Box-Behnken statistical design and the biodegradation was monitored by HPLC analysis at different time intervals. Box-Behnken results indicated that optimal conditions for biodegradation were at pH 4 and two fungal discs (10 mm diameter) in the presence of 61.2 mg L -1 of imidacloprid. A. terreus YESM3 strain was capable of degrading 85% of imidacloprid 25 mg L -1 in Czapek Dox broth medium at pH 4 and 28°C for 6 days under static conditions. In addition, after 20 days of inoculation, biodegradation recorded 96.23% of 25 mg L -1 imidacloprid. Degradation kinetics showed that the imidacloprid followed the first order kinetics with half-life (t 50 ) of 1.532 day. Intermediate product identified as 6-chloronicotinic acid (6CNA) as one of the major metabolites during degradation of imidacloprid by using HPLC. Thus, A. terreus YESM3 showed a potential to reduce pollution by pesticides and toxicity in the effected environment. However, further studies should be conducted to understand the biodegradation mechanism of this pesticide in liquid media.

  19. Effect of Essential Oils of Syzygium aromaticum and Melaleuca alternifolia on Isolates of Aspergillus sp.

    Directory of Open Access Journals (Sweden)

    A. Lima

    2014-03-01

    Full Text Available Essential  oils  have  antimicrobial  substances,  lower  cost  and  the  lower  resistance  of microorganism. This study aimed to evaluate the inhibition of mycelial growth of Aspergillus flavus and  A.  niger  with  essential  oils  of  Syzygium  aromaticum  and  Melaleuca  alternifolia.  For  this, aliquots  (0.5,  5,  10  and  15μL  of  essential  oils  from  S.  aromaticum  and  M.  alternifolia   were distributed on the surface of the culture medium Potato Dextrose Agar (PDA with Drigaslsky. In the control treatment were used only plates containing PDA plus chloramphenicol (1%. After 2 hours, a disc (8mm diameter of the isolated mycelium, with 10 days of age on PDA, was peaked to the center of the plates, these plates were sealed and incubated at 282°C, in the dark. The essential  oil  of  S.  aromaticum  inhibited  the  mycelial  growth  of  A.  flavus  and  A.  niger,  in  all aliquots. When using the essential oil of M. alternifolia decreased the mycelial growth of A. flavus in all aliquots, however, showed a low efficiency in control of A. niger. It was concluded that the essential  oil  S.  aromaticum  is  effective  against  A.  flavus  and  A.  niger,  which  can  be  used  in control against these microorganisms, and M. alternifolia not show satisfactory results in relation to reducing the growth of pathogens evaluated.

  20. Morphological Characterization and Determination of Aflatoxin-Production Potentials of Aspergillus flavus Isolated from Maize and Soil in Kenya

    Directory of Open Access Journals (Sweden)

    Matome Gabriel Thathana

    2017-09-01

    Full Text Available This study aimed at morphologically identifying Aspergillus flavus in soil and maize and at determining their aflatoxin-producing potentials. Five hundred and fourteen isolates obtained from maize and soil in Kenya were cultivated on Czapeck Dox Agar, Malt Extract Agar, Sabouraud Dextrose Agar, Potato Dextrose Agar, and Rose-Bengal Chloramphenicol Agar. Isolates were identified using macro-morphological characteristics. Micromorphological characteristics were determined using slide cultures. Aflatoxin production was determined by direct visual determination of the UV fluorescence of colonies on Coconut Agar Medium, Yeast Extract Sucrose agar, and Yeast Extract Cyclodextrin Sodium Deoxycholate agar and by Thin Layer Chromatography. Forty-three presumptive A. flavus isolates were identified; aflatoxin was detected in 23% of the isolates by UV fluorescence screening and in 30% by Thin-Layer Chromatography (TLC. The aflatoxins produced were: aflatoxin B1 (AFB1, aflatoxin B2 (AFB2, and aflatoxin G1 (AFG1; some isolates produced only AFB1, whereas others produced either AFB1 and AFB2 or AFB1 and AFG1. The highest incidence of A. flavus (63% and aflatoxin production (28% was recorded in samples from Makueni District. Isolates from Uasin Gishu (21% and Nyeri (5% were non-aflatoxigenic. Bungoma District recorded 11% positive isolates of which 2% were aflatoxin producers. The occurrence of aflatoxin-producing A. flavus emphasises the need for measures to eliminate their presence in food crops.

  1. Isolation of a Seawater Tolerant Leptospira spp. from a Southern Right Whale (Eubalaena australis).

    Science.gov (United States)

    Grune Loffler, Sylvia; Rago, Virginia; Martínez, Mara; Uhart, Marcela; Florin-Christensen, Monica; Romero, Graciela; Brihuega, Bibiana

    2015-01-01

    Leptospirosis is the most widespread zoonotic disease in the world. It is caused by pathogenic spirochetes of the genus Leptospira spp. and is maintained in nature through chronic renal infection of carrier animals. Rodents and other small mammals are the main reservoirs. Information on leptospirosis in marine mammals is scarce; however, cases of leptospirosis have been documented in pinniped populations from the Pacific coast of North America from southern California to British Columbia. We report the isolation of a Leptospira spp. strain, here named Manara, from a kidney sample obtained from a Southern Right Whale (Eubalaena australis) calf, which stranded dead in Playa Manara, Península Valdés, Argentina. This strain showed motility and morphology typical of the genus Leptospira spp. under dark-field microscopy; and grew in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium and Fletcher medium after 90 days of incubation at 28°C. Considering the source of this bacterium, we tested its ability to grow in Fletcher medium diluted with seawater at different percentages (1%, 3%, 5%, 7% and 10% v/v). Bacterial growth was detected 48 h after inoculation of Fletcher medium supplemented with 5% sea water, demonstrating the halophilic nature of the strain Manara. Phylogenetic analysis of 16S rRNA gene sequences placed this novel strain within the radiation of the pathogenic species of the genus Leptospira spp., with sequence similarities within the range 97-100%, and closely related to L. interrogans. Two different PCR protocols targeting genus-specific pathogenic genes (G1-G2, B64I-B64II and LigB) gave positive results, which indicates that the strain Manara is likely pathogenic. Further studies are needed to confirm this possibility as well as determine its serogroup. These results could modify our understanding of the epidemiology of this zoonosis. Until now, the resistance and ability to grow in seawater for long periods of time had been proven for the strain

  2. Isolation of a Seawater Tolerant Leptospira spp. from a Southern Right Whale (Eubalaena australis.

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    Sylvia Grune Loffler

    Full Text Available Leptospirosis is the most widespread zoonotic disease in the world. It is caused by pathogenic spirochetes of the genus Leptospira spp. and is maintained in nature through chronic renal infection of carrier animals. Rodents and other small mammals are the main reservoirs. Information on leptospirosis in marine mammals is scarce; however, cases of leptospirosis have been documented in pinniped populations from the Pacific coast of North America from southern California to British Columbia. We report the isolation of a Leptospira spp. strain, here named Manara, from a kidney sample obtained from a Southern Right Whale (Eubalaena australis calf, which stranded dead in Playa Manara, Península Valdés, Argentina. This strain showed motility and morphology typical of the genus Leptospira spp. under dark-field microscopy; and grew in Ellinghausen-McCullough-Johnson-Harris (EMJH medium and Fletcher medium after 90 days of incubation at 28°C. Considering the source of this bacterium, we tested its ability to grow in Fletcher medium diluted with seawater at different percentages (1%, 3%, 5%, 7% and 10% v/v. Bacterial growth was detected 48 h after inoculation of Fletcher medium supplemented with 5% sea water, demonstrating the halophilic nature of the strain Manara. Phylogenetic analysis of 16S rRNA gene sequences placed this novel strain within the radiation of the pathogenic species of the genus Leptospira spp., with sequence similarities within the range 97-100%, and closely related to L. interrogans. Two different PCR protocols targeting genus-specific pathogenic genes (G1-G2, B64I-B64II and LigB gave positive results, which indicates that the strain Manara is likely pathogenic. Further studies are needed to confirm this possibility as well as determine its serogroup. These results could modify our understanding of the epidemiology of this zoonosis. Until now, the resistance and ability to grow in seawater for long periods of time had been proven

  3. Gene expression profiles of Aspergillus flavus isolates responding to oxidative stress in different culture media

    Science.gov (United States)

    Aflatoxin contamination of peanut by Aspergillus flavus is exacerbated by drought stress. Drought also stimulates the production of reactive oxygen species (ROS) in plant tissues implying a correlation between ROS and aflatoxin production. Here, we performed gene expression analysis by RNAseq of tox...

  4. Comparative transcriptome analysis of Aspergillus flavus isolates under different oxidative stresses and culture media

    Science.gov (United States)

    Aspergillus flavus and aflatoxin contamination in the field are known to be influenced by numerous stress factors, particularly drought and heat stress. However, the purpose of aflatoxin production is unknown. Here, we report transcriptome analyses comprised of 282.6 Gb of sequencing data describing...

  5. Seed isolates of Alternaria and Aspergillus fungi increase germination of Astragalus utahensis

    Science.gov (United States)

    Sean D. Eldredge; Brad Geary; Scott L. Jensen

    2016-01-01

    Astragalus utahensis (Torr.) Torr. & A. Gray (Fabaceae) (Utah milkvetch) is native lo the arid Great Basin and has desirable attributes that make it a good candidate for restoration in arid, noncompetitive situations. Seed dormancy is a significant barrier to consistent establishment for this species. Species of Alternaria and Aspergillus fungi have...

  6. Biosynthesis of silver nanoparticles by Pseudomonas spp. isolated from effluent of an electroplating industry.

    Science.gov (United States)

    Punjabi, Kapil; Yedurkar, Snehal; Doshi, Sejal; Deshapnde, Sunita; Vaidya, Shashikant

    2017-08-01

    The aim of this study was to isolate and screen bacteria from soil and effluent of electroplating industries for the synthesis of silver nanoparticles and characterize the potential isolate. Soil and effluent of electroplating industries from Mumbai were screened for bacteria capable of synthesizing silver nanoparticles. From two soils and eight effluent samples 20 bacterial isolates were obtained, of these, one was found to synthesize silver nanoparticles. Synthesis of silver nanoparticle by bacteria was confirmed by undertaking characterization studies of nanoparticles that involved spectroscopy and electron microscopic techniques. The potential bacteria was found to be Gram-negative short rods with its biochemical test indicating Pseudomonas spp . Molecular characterization of the isolate by 16S r DNA sequencing was carried out which confirmed its relation to Pseudomonas hibiscicola ATCC 19867. Stable nanoparticles synthesized were 50 nm in size and variable shapes as seen in SEM micrographs. The XRD and FTIR confirmed the crystalline structure of nanoparticles and presence of biomolecules mainly proteins as agents for reduction and capping of nanoparticles. The study demonstrates synthesis of nanoparticles by bacteria from effluent of electroplating industry. This can be used for large scale synthesis of nanoparticles by cost effective and environmentally benign mode of synthesis.

  7. [Bacillus isolates from rhizosphere of cacti improve germination and bloom in Mammillaria spp. (Cactaceae)].

    Science.gov (United States)

    Chávez-Ambriz, Lluvia A; Hernández-Morales, Alejandro; Cabrera-Luna, José A; Luna-Martínez, Laura; Pacheco-Aguilar, Juan R

    Cacti are the most representative vegetation of arid zones in Mexico where rainfall is scarce, evapotranspiration is high and soil fertility is low. Plants have developed physiological strategies such as the association with microorganisms in the rhizosphere zone to increase nutrient uptake. In the present work, four bacterial isolates from the rhizosphere of Mammillaria magnimamma and Coryphantha radians were obtained and named as QAP3, QAP19, QAP22 and QAP24, and were genetically identified as belonging to the genus Bacillus, exhibiting in vitro biochemical properties such as phosphate solubilization, indoleacetic acid production and ACC deaminase activity related to plant growth promotion, which was tested by inoculating M. magnimamma seeds. It was found that all isolates increased germination from 17 to 34.3% with respect to the uninoculated control seeds, being QAP24 the one having the greatest effect, accomplishing the germination of viable seeds (84.7%) three days before the control seeds. Subsequently, the inoculation of Mammillari zeilmanniana plants with this isolate showed a positive effect on bloom, registering during two months from a one year period, an increase of up to 31.0% in the number of flowering plants compared to control plants. The characterized Bacillus spp. isolates have potential to be used in conservation programs of plant species from arid zones. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  8. Identification and characterization of novel natural pathogen of Drosophila melanogaster isolated from wild captured Drosophila spp.

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    Singh, Karan; Zulkifli, Mohammad; Prasad, N G

    2016-12-01

    Drosophila melanogaster is an emerging model system for the study of evolutionary ecology of immunity. However, a large number of studies have used non natural pathogens as very few natural pathogens have been isolated and identified. Our aim was to isolate and characterize natural pathogen/s of D. melanogaster. A bacterial pathogen was isolated from wild caught Drosophila spp., identified as a new strain of Staphylococcus succinus subsp. succinus and named PK-1. This strain induced substantial mortality (36-62%) in adults of several laboratory populations of D. melanogaster. PK-1 grew rapidly within the body of the flies post infection and both males and females had roughly same number of colony forming units. Mortality was affected by mode of infection and dosage of the pathogen. However mating status of the host had no effect on mortality post infection. Given that there are very few known natural bacterial pathogens of D. melanogaster and that PK-1 can establish a sustained infection across various outbred and inbred populations of D. melanogaster this new isolate is a potential resource for future studies on immunity. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  9. Serotypes and Antimicrobial Susceptibility of Salmonella spp. Isolated from Farm Animals in China

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    Yuan Zong Hui

    2015-06-01

    Full Text Available Salmonella spp. can indirectly infect humans via transfer from animals and animal-derived food products, and thereby cause potentially fatal diseases. Therefore, gaining an understanding of Salmonella infection in farm animals is increasingly important. The aim of this study was to identify the distribution of serotypes in Salmonella samples isolated from chickens (n = 837, pigs (n = 930, and dairy cows (n = 418 in central China (Henan, Hubei, and Hunan provinces in 2010–2011, and investigate the susceptibility of strains to antimicrobial agents. Salmonella isolates were identified by PCR amplification of the invA gene, serotypes were determined by using a slide agglutination test for O and H antigens, and susceptibility to 24 antimicrobials was tested using the agar dilution method. In total, 248 Salmonella strains were identified: 105, 105, and 38 from chickens, dairy cows, and pigs, respectively. Additionally, 209 strains were identified in unhealthy pigs from the Huazhong Agricultural University veterinary hospital. Among these 457 strains, the dominant serotypes were Typhimurium in serogroup B, IIIb in serogroup C, and Enteritidis in serogroup D. In antimicrobial susceptibility tests, 41.14% of Salmonella spp. were susceptible to all antimicrobial agents, 48.14% were resistant to at least one, and 34.72% were resistant to more than three classes. Strains were highly resistant to sulfamethoxazole-trimethoprim (39.61%, nalidixic acid (39.17%, doxycycline (28.22%, and tetracycline (27.58%. Resistance to cephalosporins and fluoroquinolones ranged from 5.25% to 7.44% and 19.04% to 24.51%, respectively. Among penicillin-resistant and cephalosporin-resistant strains, 25 isolates produced extended-spectrum β-lactamases (ESBLs. The multidrug-resistant and ESBL-producing Salmonella strains identified in healthy animals here will present a challenge for veterinary medicine and farm animal husbandry, and could also pose a threat to public health

  10. Virulence factors genes of Staphylococcus spp. isolated from caprine subclinical mastitis.

    Science.gov (United States)

    Salaberry, Sandra Renata Sampaio; Saidenberg, André Becker Simões; Zuniga, Eveline; Melville, Priscilla Anne; Santos, Franklin Gerônimo Bispo; Guimarães, Ednaldo Carvalho; Gregori, Fábio; Benites, Nilson Roberti

    2015-08-01

    The aim of this study was to investigate genes involved in adhesion expression, biofilm formation, and enterotoxin production in isolates of Staphylococcus spp. from goats with subclinical mastitis and associate these results with the staphylococcal species. One hundred and twenty-four isolates were identified and polymerase chain reaction (PCR) was performed to detect the following genes: cna, ebpS, eno, fib, fnbA, fnbB, bap, sea, seb, sec, sed and see. The most commonly Staphylococcus species included S. epidermidis, S. lugdunensis, S. chromogenes, S. capitis ss capitis and S. intermedius. With the exception of fnbB, the genes were detected in different frequencies of occurrence in 86.3% of the Staphylococcus spp. isolates. Eno (73.2%) and bap (94.8%) were more frequently detected in coagulase-negative staphylococci (CNS); ebpS (76%), fib (90.9%) and fnbA (87%) were the most frequent genes in coagulase-positive staphylococci (CPS). Regarding enterotoxins, genes sed (28.2%) and see (24.2%) had a higher frequency of occurrence; sec gene was more frequently detected in CPS (58.8%). There was no association between the presence of the genes and the Staphylococcus species. Different virulence factors genes can be detected in caprine subclinical mastitis caused by CNS and CPS. The knowledge of the occurrence of these virulence factors is important for the development of effective control and prevention measures of subclinical mastitis caused by CNS and CPS in goats. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. Antimicrobial Susceptibility and Genotypic Characteristic of Campylobacter spp. Isolates from Free-Living Birds in Poland.

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    Krawiec, Marta; Woźniak-Biel, Anna; Bednarski, Michał; Wieliczko, Alina

    2017-11-01

    Campylobacter spp. is the most commonly reported, bacterial cause of human foodborne infection worldwide. Commercial poultry and free-living birds are natural reservoirs of three particular species: Campylobacter jejuni, Campylobacter coli, and Campylobacter lari. The aim of this study was to determine the genotypic characteristics and antibiotic susceptibility of 43 Campylobacter strains, obtained from free-living birds, in Poland. In total, 700 birds were examined. The strains were isolated from 43 birds (6.14%) from the feces of 7 wild bird species: Mallard ducks Anas platyrhynchos (29 positive/121 tested), great cormorants Phalacrocorax carbo (5/77), velvet scoters Melanitta fusca (4/30), tawny owls Strix aluco (2/5), common buzzard Buteo buteo (1/3), rook Corvus frugilegus (1/6), and Eurasian tree sparrow Passer montanus (1/30). Thirty-eight (88.37%) of obtained strains belonged to C. jejuni and five (11.63%) to C. coli. Other 428 examined birds from different bird species were Campylobacter negative. The antimicrobial susceptibility to nine antimicrobials was also studied in investigated isolates of Campylobacter spp. Sixteen of the examined strains (37.21% of all positive samples) showed susceptibility to all of the nine antimicrobials. Moreover, the prevalence of selected virulence genes, such as flaA, cadF, ceuE, virB11, cdtA, cdtB, and cdtC were all analyzed. The virulence gene that was found most frequently in total number of Campylobacter strains was ceuE (72.10%) and other genes, such as flaA, cadF, cdtA, cdtB, and cdtC, were found in over 60% of all examined strains. Variable antimicrobial susceptibility and the presence of different virulence genes of examined strains, isolated from free-living birds, suggest that special attention should be given to wild birds and any potential approaches to the control of antibiotic-resistant Campylobacter should be discussed.

  12. Identification of Brucella spp. isolated from human brucellosis in Malaysia using high-resolution melt (HRM) analysis.

    Science.gov (United States)

    Mohamed Zahidi, Jama'ayah; Bee Yong, Tay; Hashim, Rohaidah; Mohd Noor, Azura; Hamzah, Siti Hawa; Ahmad, Norazah

    2015-04-01

    Molecular approaches have been investigated to overcome difficulties in identification and differentiation of Brucella spp. using conventional phenotypic methods. In this study, high-resolution melt (HRM) analysis was used for rapid identification and differentiation of members of Brucella genus. A total of 41 Brucella spp. isolates from human brucellosis were subjected to HRM analysis using 4 sets of primers, which identified 40 isolates as Brucella melitensis and 1 as Brucella canis. The technique utilized low DNA concentration and was highly reproducible. The assay is shown to be a useful diagnostic tool, which can rapidly differentiate Brucella up to species level. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  13. Prevalence and identification by multiplex polymerase chain reaction patterns of Cronobacter spp. isolated from plant-based foods

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    Filiz AKSU

    2016-01-01

    Full Text Available Abstract Cronobacter spp. involves a group of opportunistic pathogens that cause meningitis in newborns, immunosuppressed individuals with a mortality rate of 50-80%. Seven species like C. sakazakii, C. malonaticus, C. muytjensii, C. turicensis, C. dublinensis, C. universalis, C. condimenti are included in this genus which has been a subject of research especially in the bacteriologic analysis of baby foods. However, since these species were detected also in prepared foodstuffs. The objective of this study was to assert the presence of Cronobacter spp. in foodstuff offered for sale in Turkey. A total of 151 prepared foodstuffs including a variety of spice, flour, instant soup were purchased from different sales points. The presence of Cronobacter spp. were investigated in these samples. Cronobacter suspected isolates which were obtained by microbiological analyses were confirmed by PCR targeted to gyrB gene and were then identified by multiplex PCR. Prevalence of Cronobacter spp was estimated to be 17.88%. Out of 27 Cronobacter spp. isolates obtained, 13(48.1%, 6(22.2%, 5(18.5%, 3(11.1% belonged to C. sakazakii, C. muytjensii, C. turicensis, C. malonaticus species, respectively. Consequently, the presence of the bacteria in widely consumed foodstuff revealed that Cronobacter spp. is subject to monitoring due to its opportunistic nature in terms of public health concern.

  14. Fungi isolated from Stewartia pseudocamellia Max. seeds and their pathogenesis

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    Halina Kurzawińska

    2012-12-01

    Full Text Available The aim of studies was to determine typical composition of fungi occurring on seeds of Stewartia pseudocamellia.The studies conducted on 100 disinfected and 100 nondisinfected seeds of these plants.Isolates of Alternaria alternata, Fusarium oxysporum, Cylindrocarpon radicicola and Rhizoctonia solani were characterized by pathogenicity towards the investigated Stewartia pseudocamellia. In the laboratory experiment, 204 isolations of microorganisms were obtained that belonged to 20 species and form of fungi and bacteria. Among fungi there were both of parasite (Alternaria alternata, Botrytis cinerea, Fusarium spp., Rhizoctonia solani and typical saprophytic (Cladosporium spp., Penicillium spp., Aspergillus spp., Epicoccum spp., Mucor spp.. The dominant fungus on seeds was Alternaria alternata. Among the investigated isolates only one isolate (R4 Rhizoctonia solani, was strongly pathogenic, isolates (A1 Alternaria alternata were weakly pathogenic to seedlings of Stewartia pseudocamellia.

  15. MICROSCOPIC FUNGI ISOLATED FROM POLISH HONEY

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    Soňa Felšöciová

    2012-12-01

    Full Text Available The characterization of some honey samples from Poland was carried out on the basis of their microbiological (fungi and yeasts analysis. Most of the samples contained less than 20 % water. The amount of fungi found in the honey samples was less than 1 x 102 CFU.g-1 but 19 % of the samples had more yeasts than 1 x 102 CFU.g-1 – up to 5.7 x 102 CFU.g-1. The isolated fungi were Alternaria spp., Aspergillus spp., Cladosporium spp., Fusarium spp., Mycelia sterilia, Rhizopus spp. and Penicillium spp. The last genus was isolated very frequently. A total number of eight fungal Penicillium species were identified namely, Penicillium brevicompactum, P. commune, P. corylophilum, P. crustosum, P. expansum, P. griseofulvum, P. chrysogenum and P. polonicum. They were isolated using dilution plate method. The results showed that honeys produced in this region are of good microbiological quality.

  16. Genetic Diversity and Differentiation of Colletotrichum spp. Isolates Associated with Leguminosae Using Multigene Loci, RAPD and ISSR

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    Farshid Mahmodi

    2014-03-01

    Full Text Available Genetic diversity and differentiation of 50 Colletotrichum spp. isolates from legume crops studied through multigene loci, RAPD and ISSR analysis. DNA sequence comparisons by six genes (ITS, ACT, Tub2, CHS-1, GAPDH, and HIS3 verified species identity of C. truncatum, C. dematium and C. gloeosporiodes and identity C. capsici as a synonym of C. truncatum. Based on the matrix distance analysis of multigene sequences, the Colletotrichum species showed diverse degrees of intera and interspecific divergence (0.0 to 1.4% and (15.5–19.9, respectively. A multilocus molecular phylogenetic analysis clustered Colletotrichum spp. isolates into 3 well-defined clades, representing three distinct species; C. truncatum, C. dematium and C. gloeosporioides. The ISSR and RAPD and cluster analysis exhibited a high degree of variability among different isolates and permitted the grouping of isolates of Colletotrichum spp. into three distinct clusters. Distinct populations of Colletotrichum spp. isolates were genetically in accordance with host specificity and inconsistent with geographical origins. The large population of C. truncatum showed greater amounts of genetic diversity than smaller populations of C. dematium and C. gloeosporioides species. Results of ISSR and RAPD markers were congruent, but the effective maker ratio and the number of private alleles were greater in ISSR markers.

  17. Isolation and identification of fungal flora associated with groundnut ...

    African Journals Online (AJOL)

    A total of 25 colonies were isolated from all the samples from which 6 fungal species were identified, namely Mucor, Aspergillus, Rhizophus, Curvularia, Pencillium and Fusarium spp. Of these, Mucor and Rhizopus were most prevalent having been isolated from the three storage facilities studied. Curvularia and Penicillium ...

  18. Rapid identification of pathogenic streptococci isolated from moribund red tilapia (Oreochromis spp.).

    Science.gov (United States)

    Abdelsalam, Mohamed; Elgendy, Mamdouh Y; Shaalan, Mohamed; Moustafa, Mohamed; Fujino, Masayuki

    2017-03-01

    Accurate and rapid identification of bacterial pathogens of fish is essential for the effective treatment and speedy control of infections. Massive mortalities in market-sized red tilapia (Oreochromis spp.) were noticed in mariculture concrete ponds in northern Egypt. Histopathological examination revealed marked congestion in the central vein of the liver with the presence of bacterial aggregates inside the lumen and in the vicinity of the central vein. A total of 12 isolates of streptococci were obtained from the moribund fish. This study documented the ability of the MicroSeq 500 16S bacterial sequencing method to accurately identify Streptococcus agalactiae and S. dysgalactiae mixed infections from moribund red tilapia that were difficult to be recognised by the commercial biochemical systems. The continuously decreasing cost of the sequencing technique should encourage its application in routine diagnostic procedures.

  19. Isolation of Campylobacter spp. from Three Species of Antarctic Penguins in Different Geographic Locations.

    Science.gov (United States)

    García-Peña, F J; Llorente, M T; Serrano, T; Ruano, M J; Belliure, J; Benzal, J; Herrera-León, S; Vidal, V; D'Amico, V; Pérez-Boto, D; Barbosa, A

    2017-03-01

    The presence of Campylobacter species was studied in three Antarctic penguin species, Adélie (Pygoscelis adeliae), chinstrap (Pygoscelis antarctica) and gentoo (Pygoscelis papua). A total of 390 penguins were captured in 12 different rookeries along the Antarctic Peninsula with differences in the amount of human visitation: six colonies were highly visited [Stranger Point, King George Island (P. papua and P. adeliae); Hannah Point, Livingston Island (P. papua and P. antarctica); Deception Island (P. antarctica); and Paradise Bay, Antarctic Peninsula (P. papua)], and six colonies were rarely visited [Devil's Point, Byers Peninsula, Livingston Island (P. papua); Cierva Cove, Antarctic Peninsula (P. papua); Rongé Island (P. papua and P. antarctica); Yalour Island (P. adeliae); and Avian Island (P. adeliae)]. A total of 23 strains were isolated from penguins from nine different rookeries. Campylobacter lari subsp. lari was isolated from eight samples (seven from P. papua and one from P. adeliae); C. lari subsp. concheus from 13 (ten from P. adeliae and three from P. antarctica) and C. volucris from two samples (both from P. papua). We did not find any significant differences in the prevalence of Campylobacter spp. between the populations in highly and rarely visited areas. This is the first report of C. lari subsp. concheus and C. volucris isolation from penguins in the Antarctic region.

  20. Genotypes of Leptospira spp. strains isolated from dogs in Buenos Aires, Argentina.

    Science.gov (United States)

    Grune Loffler, Sylvia; Passaro, Diego; Samartino, Luis; Soncini, Analía; Romero, Graciela; Brihuega, Bibiana

    2014-01-01

    Leptospirosis is an infectious disease of wide global distribution, which is endemic in Argentina. The objective of this study was to obtain the genetic profiles of Leptospira spp. strains isolated from clinical cases of dogs in the province of Buenos Aires by the multiple-locus variable-number tandem repeat analysis (MLVA). Eight isolated canine strains were genotyped by MLVA, obtaining the identical profile of Leptospira interrogans serovar Canicola Hond Utrecht IV in the strains named Dogy and Mayo. The strains named Bel, Sarmiento, La Plata 4581 and La Plata 5478 were identical to the profile of the genotype of L. interrogans serovar Portlandvere MY 1039.The strain named Avellaneda was identical to the genotype profile of L. interrogans serovar Icterohaemorrhagiae RGA and the strain named SB had the same profile as the L. interrogans serovar Pomona Baires genotype and was similar to the profile of serovar Pomona Pomona genotype. It would be useful to include a larger number of isolates from different dog populations in various provinces of Argentina and to characterize the genetic profiles of the strains circulating in the country. The information obtained will be useful for the control of leptospirosis in the dog population. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España. All rights reserved.

  1. The potential of compounds isolated from Xylaria spp. as antifungal agents against anthracnose.

    Science.gov (United States)

    Elias, Luciana M; Fortkamp, Diana; Sartori, Sérgio B; Ferreira, Marília C; Gomes, Luiz H; Azevedo, João L; Montoya, Quimi V; Rodrigues, André; Ferreira, Antonio G; Lira, Simone P

    2018-03-31

    Anthracnose is a crop disease usually caused by fungi in the genus Colletotrichum or Gloeosporium. These are considered one of the main pathogens, causing significant economic losses, such as in peppers and guarana. The current forms of control include the use of resistant cultivars, sanitary pruning and fungicides. However, even with the use of some methods of controlling these cultures, the crops are not free of anthracnose. Additionally, excessive application of fungicides increases the resistance of pathogens to agrochemicals and cause harm to human health and the environment. In order to find natural antifungal agents against guarana anthracnose, endophytic fungi were isolated from Amazon guarana. The compounds piliformic acid and cytochalasin D were isolated by chromatographic techniques from two Xylaria spp., guided by assays with Colletotrichum gloeosporioides. The isolated compounds were identified by spectrometric techniques, as NMR and mass spectrometry. This is the first report that piliformic acid and cytochalasin D have antifungal activity against C. gloeosporioides with MIC 2.92 and 2.46μmolmL -1 respectively. Captan and difenoconazole were included as positive controls (MIC 16.63 and 0.02μmolmL -1 , respectively). Thus, Xylaria species presented a biotechnological potential and production of different active compounds which might be promising against anthracnose disease. Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  2. Genetic diversity of thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina.

    Science.gov (United States)

    Zbrun, María V; Romero-Scharpen, Analía; Olivero, Carolina; Zimmermann, Jorge A; Rossler, Eugenia; Soto, Lorena P; Astesana, Diego M; Blajman, Jesica E; Berisvil, Ayelén; Frizzo, Laureano S; Signorini, Marcelo L

    The objective of this study was to investigate a clonal relationship among thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina. A total of 128 thermotolerant Campylobacter spp. (89 C. jejuni and 39 C. coli) isolates from six poultry meat chains were examined. These isolates were from: a) hens from breeder flocks, b) chickens on the farm (at ages 1 wk and 5 wk), c) chicken carcasses in the slaughterhouse, and d) chicken carcasses in the retail market. Chickens sampled along each food chain were from the same batch. Campylobacter spp. isolates were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Clustering of C. jejuni isolates resulted in 17 profiles, with four predominant genotypes and many small profiles with just a few isolates or unique patterns, showing a very high degree of heterogeneity among the C. jejuni isolates. Some clusters included isolates from different stages within the same chain, which would indicate a spread of strains along the same poultry meat chain. Moreover, twenty-two strains of C. coli clustered in seven groups and the remaining 17 isolates exhibited unique profiles. Evidence for transmission of thermotolerant Campylobacter spp. through the food chain and cross contamination in the slaughterhouses were obtained. This collective evidence should be considered as the scientific basis to implement risk management measures to protect the public health. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  3. First isolation of a novel thermostable antifungal peptide secreted by Aspergillus clavatus.

    Science.gov (United States)

    Skouri-Gargouri, Houda; Gargouri, Ali

    2008-11-01

    A novel antifungal peptide produced by an indigenous fungal strain (VR) of Aspergillus clavatus was purified. The antifungal peptide was enriched in the supernatant after heat treatment at 70 degrees C. The thermostable character was exploited in the first purification step, as purified peptide was obtained after ultrafiltration and reverse phase-HPLC on C18 column application. The purified peptide named "AcAFP" for A. clavatus antifungal peptide, has molecular mass of 5773Da determined by MALDI-ToF spectrometry. The N-terminal sequence showed a notable identity to the limited family of antifungal peptides produced by ascomycetes fungi. The AcAFP activity remains intact even after heat treatment at 100 degrees C for 1h confirming its thermostability. It exhibits a strong inhibitory activity against mycelial growth of several serious human and plant pathogenic fungi: Fusariuym oxysporum, Fusarium solani, Aspergillus niger, Botrytis cinerea, Alternaria solani, whereas AcAFP did not affect yeast and bacterial growth.

  4. Isolation of lipase producing fungi from palm oil Mill effluent (POME dump sites at Nsukka

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    Charles Ogugua Nwuche

    2011-02-01

    Full Text Available In this study, twelve fungal lipase producing strains belonging to Aspergillus, Penicillium, Trichoderma and Mucor genera were isolated from palm oil mill effluent composts. The Aspergillus spp. were more frequent (42% and was present in all the samples assayed. Mucor sp. was the least encountered (8.3%.The lipase producing profile showed that Trichoderma (8.07-8.24 u/mL and Aspergillus (6.25 -7.54 u/mL spp. were the highest lipase producers while Mucor (5.72 u/mL was the least.

  5. Polyphasic identification of Aspergillus isolates belonging to section Nigri with clinical relevance

    OpenAIRE

    Maciel, Marília; Santos, Cledir; Lima, Nelson; Souza-Motta, Cristina Maria

    2012-01-01

    Aspergillosis is the name of a group of diseases in humans and animals caused by opportunist moulds of the genus Aspergillus. The vast majority of infections are caused by A. fumigatus, followed by other species such as A. flavus, A. terreus and A. niger. Among the pulmonary infection, aspergillosis is gaining prominent position not only in immunocompromised patients, but also in immunosuppressed. The absence of a reliable fungal identification system affects the control of ...

  6. Molecular and biochemical characterization of soil isolates of Aspergillus niger aggregate and an assessment of their antagonism against Rhizoctonia solani

    Directory of Open Access Journals (Sweden)

    M.R. Khan

    2007-12-01

    Full Text Available Sixteen Aspergillus niger aggregate isolates collected from different crop fi elds were subjected to RAPDPCR using 20 Operon primers and 8 synthetic primers. Twenty-two primers led to the amplifi cation of 727 fragments ranging from 3500 bp (OPA 11 to 200 bp (primer 06. Two bands were monomorphic, while the rest were polymorphic. Three amplicons produced by OPA 16 were recorded as isolate-specifi c as 2300 bp by AnC2 and AnR3, and as 2800 bp by AnC2 only. The highest genetic similarity (0.79 was measured between AnC2 and AnR3, and the lowest (0.17 between AnC2 and AnR2. Multivariate analysis of genetic similarity revealed three major clusters, named group I, group II and group III. All isolates were ochratoxin A negative (<1 ng g-1. Isolates AnC2 and AnR3, which produced HCN and solubilized the greatest amount of phosphorus, belonged to group I. These isolates also signifi cantly increased eggplant yield and caused the greatest inhibition of colonization by R. solani in dual culture. They also suppressed the root rot on eggplant and the soil population of R. solani in pot soil.

  7. Isolation and optimization of pectinase enzyme production one of useful industrial enzyme in Aspergillus niger, Rhizopus oryzae, Penicilium chrysogenum

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    akram songol

    2016-06-01

    Full Text Available Introduction: Pectinase enzyme is one of the most important industrial enzymes which isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the fruit and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi performed through plate culture on pectin medium and staining with Lugol's iodine solution. The best strains were identified by ITS1, 4 sequencing as Aspergillus fumigatus, Rhizopus oryzae, Penicilium chrysogenum. The enzyme production was optimized by application of the five factorial design, each at three levels. These factors are carbon sources (whey, glucose and stevia, ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results indicate that optimum condition for enzyme production for three fungi strains was obtained at 32 °C, pH = 6, 3g / L manganese sulfate, 2.75g / L of ammonium sulfate and 10g / L of each carbon source. The best experiment in obtaining the optimum enzyme contained 1.328 mg / ml of glucose for Aspergillus niger 1.284 and 1.039 mg / ml of whey for Rhizopus oryzae and Penicilium chrysogenum. Molecular weight of enzyme was about 40 and 37 kDa which was obtained by SDS- PAGE. Discussion and conclusion: The results indicate that three strains could grow in a wide range of carbon source, pH and temperature, which could be a good candidate for industrial application.

  8. Typing clinical and animal environment Aspergillus fumigatus gliotoxin producer strains isolated from Brazil by PCR-RFLP markers.

    Science.gov (United States)

    Soleiro, C A; Pena, G A; Cavaglieri, L R; Coelho, I; Keller, L M; Dalcero, A M; Rosa, C A R

    2013-12-01

    Aspergillus fumigatus, a well-known human and animal pathogen causing aspergillosis, has been historically identified by morphological and microscopic features. However, recent studies have shown that species identification on the basis of morphology alone is problematic. The aim of this work was to confirm the taxonomic state at specie level of a set of clinical (human and animal) and animal environment A. fumigatus strains identified by morphological criteria applying a PCR-RFLP assay by an in silico and in situ analysis with three restriction enzymes. The A. fumigatus gliotoxin-producing ability was also determined. Previous to the in situ PCR-RFLP analysis, an in silico assay with BccI, MspI and Sau3AI restriction enzymes was carried out. After that, these enzymes were used for in situ assay. All A. fumigatus strains isolated from corn silage, human aspergillosis and bovine mastitis and high per cent of the strains isolated from cereals, animal feedstuff and sorghum silage were able to produce high gliotoxin levels. Also, all these strains identified by morphological criteria as A. fumigatus, regardless of its isolation source, had band patterns according to A. fumigatus sensu stricto by PCR-RFLP markers. Aspergillus fumigatus is a well-known human and animal pathogen causing aspergillosis. In this study, clinical (human and animal) and animal environment strains were able to produce high gliotoxin levels and had band profiles according to A. fumigatus sensu stricto by PCR-RFLP markers. The results obtained here suggest that strains involved in human and animal aspergillosis could come from the animal environment in which A. fumigatus is frequently found. Its presence in animal environments could affect animal health and productivity; in addition, there are risks of contamination for rural workers during handling and storage of animal feedstuffs. © 2013 The Society for Applied Microbiology.

  9. Isolation and Characterization of Pseudomonas spp. Strains That Efficiently Decompose Sodium Dodecyl Sulfate

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    Ewa M. Furmanczyk

    2017-11-01

    Full Text Available Due to their particular properties, detergents are widely used in household cleaning products, cosmetics, pharmaceuticals, and in agriculture as adjuvants tailoring the features of pesticides or other crop protection agents. The continuously growing use of these various products means that water soluble detergents have become one of the most problematic groups of pollutants for the aquatic and terrestrial environments. Thus it is important to identify bacteria having the ability to survive in the presence of large quantities of detergent and efficiently decompose it to non-surface active compounds. In this study, we used peaty soil sampled from a surface flow constructed wetland in a wastewater treatment plant to isolate bacteria that degrade sodium dodecyl sulfate (SDS. We identified and initially characterized 36 Pseudomonas spp. strains that varied significantly in their ability to use SDS as their sole carbon source. Five isolates having the closest taxonomic relationship to the Pseudomonas jessenii subgroup appeared to be the most efficient SDS degraders, decomposing from 80 to 100% of the SDS present in an initial concentration 1 g/L in less than 24 h. These isolates exhibited significant differences in degree of SDS degradation, their resistance to high detergent concentration (ranging from 2.5 g/L up to 10 g/L or higher, and in chemotaxis toward SDS on a plate test. Mass spectrometry revealed several SDS degradation products, 1-dodecanol being dominant; however, traces of dodecanal, 2-dodecanol, and 3-dodecanol were also observed, but no dodecanoic acid. Native polyacrylamide gel electrophoresis zymography revealed that all of the selected isolates possessed alkylsulfatase-like activity. Three isolates, AP3_10, AP3_20, and AP3_22, showed a single band on native PAGE zymography, that could be the result of alkylsulfatase activity, whereas for isolates AP3_16 and AP3_19 two bands were observed. Moreover, the AP3_22 strain exhibited a band

  10. Multicenter evaluation of resistance patterns of Klebsiella pneumoniae, Escherichia coli, Salmonella spp and Shigella spp isolated from clinical specimens in Brazil: RESISTNET surveillance program

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    Carmen Paz Oplustil

    Full Text Available Surveillance programs are essential to detect the increase of antimicrobial resistance, and several different programs are being conducted in many countries. The RESISTNET is a surveillance program for bacterial resistance against several antimicrobial agents initiated in 1998 among Latin American countries. In Brazil, several centers were invited to join this surveillance and a total of 11 centers (6 from São Paulo and 5 from other states participated in the study. All results were analyzed using the WHONET program. A total of 894 Escherichia coli, 386 Klebsiella pneumoniae, 70 Shigella spp and 57 Salmonella spp strains were analyzed in this study from April, 1998, to April, 1999. Susceptibility testing was performed by the disk diffusion method using NCCLS 1998 guidelines for several different drugs. For all strains, imipenem was the most effective drug (100% of the strains were susceptible. Klebsiella pneumoniae presented a high resistance rate to ampicillin (96.4%. The rate of probable ESBL producers among K. pneumoniae strains was 36.3%, most of them being isolated from catheters (58.8%. Among all Escherichia coli strains analyzed, the highest resistance rate was found for trimethoprim/sulfamethoxazole (46.9% and the majority of the resistant strains were isolated from urine samples (47.8%. Among Salmonella spp, the resistance rates were low for all antibiotics tested. For Shigella spp strains there was a high resistance to trimethoprim/sulfamethoxazole (80.0%. No resistance to ceftriaxone was observed in these strains. Surveillance of antimicrobial resistance is critical for the successful management of infectious diseases. The results of this survey show significant resistance rates among these bacteria which are responsible for several types of human infections.

  11. Variation in fumonisin and ochratoxin production associated with differences in biosynthetic gene content in Aspergillus niger and A. welwitschiae isolates from multiple crop and geographic origins

    Science.gov (United States)

    The fungi Aspergillus niger and A. welwitschiae are morphologically indistinguishable species used for industrial fermentation and for food and beverage production. The fungi also occur widely on food crops. Concerns about their safety have arisen with the discovery that some isolates of both specie...

  12. Comparative genomics analysis of field isolates of Aspergillus flavus and A. parasiticus to explain phenotypic variation in oxidative stress tolerance and host preference

    Science.gov (United States)

    Aflatoxin contamination of peanut and other crops is a major concern for producers globally, and has been shown to be exacerbated by drought stress. Previous transcriptomic and proteomic examination of the responses of isolates of Aspergillus flavus to drought-related oxidative stress in vitro have ...

  13. Inhibition of aflatoxin B production of Aspergillus flavus, isolated from soybean seeds by certain natural plant products.

    Science.gov (United States)

    Krishnamurthy, Y L; Shashikala, J

    2006-11-01

    The inhibitory effect of cowdung fumes, Captan, leaf powder of Withania somnifera, Hyptis suaveolens, Eucalyptus citriodora, peel powder of Citrus sinensis, Citrus medica and Punica granatum, neem cake and pongamia cake and spore suspension of Trichoderma harzianum and Aspergillus niger on aflatoxin B(1) production by toxigenic strain of Aspergillus flavus isolated from soybean seeds was investigated. Soybean seed was treated with different natural products and fungicide captan and was inoculated with toxigenic strain of A. flavus and incubated for different periods. The results showed that all the treatments were effective in controlling aflatoxin B(1) production. Captan, neem cake, spore suspension of T. harzianum, A. niger and combination of both reduced the level of aflatoxin B(1) to a great extent. Leaf powder of W. somnifera, H. suaveolens, peel powder of C. sinensis, C. medica and pongamia cake also controlled the aflatoxin B(1) production. All the natural product treatments applied were significantly effective in inhibiting aflatoxin B(1) production on soybean seeds by A. flavus. These natural plant products may successfully replace chemical fungicides and provide an alternative method to protect soybean and other agricultural commodities from aflatoxin B(1) production by A. flavus.

  14. Novel whole-cell Reporter Assay for Stress-Based Classification of Antibacterial Compounds Produced by Locally Isolated Bacillus spp.

    OpenAIRE

    Nithya, Vadakedath; Halami, Prakash M.

    2012-01-01

    Reporter bacteria are beneficial for the rapid and sensitive screening of cultures producing peptide antibiotics, which can be an addition or alternative to the established antibiotics. This study was carried out to validate the usability of specific reporter strains for the target mediated identification of antibiotics produced by native Bacillus spp. isolated from different food sources. During preliminary classification, cell wall stress causing Bacillus isolates were screened by using rep...

  15. Azasordarins: Susceptibility of Fluconazole-Susceptible and Fluconazole-Resistant Clinical Isolates of Candida spp. to GW 471558

    OpenAIRE

    Cuenca-Estrella, Manuel; Mellado, Emilia; Díaz-Guerra, Teresa M.; Monzón, Araceli; Rodríguez-Tudela, Juan L.

    2001-01-01

    The in vitro activity of the azasordarin GW 471558 was compared with those of amphotericin B, flucytosine, itraconazole, and ketoconazole against 177 clinical isolates of Candida spp. GW 471558 showed potent activity against Candida albicans, Candida glabrata, and Candida tropicalis, even against isolates with decreased susceptibility to azoles. Candida krusei, Candida parapsilosis, Candida lusitaniae, and Candida guilliermondii are resistant to GW 471558 in vitro (MICs, >128 μg/ml).

  16. Determination of some virulence factors of Candida spp. isolated from locally produced cheese in Diyala Governorate-Iraq

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    Suhail Jawdat Fadihl

    2017-03-01

    Full Text Available Locally produced cheese which called (Gibin Al arab is one of the most common dairy products in Iraq, it has an economic importance and great social value. This research aimed to identify yeast species from locally produced cheese (Gibin Al Arab in Diyala city which traditionally made and sold in markets of old town in Baquba, and study some of virulence factors (Esterase production, Phospholipase and Hemolytic production of yeasts belong to genus of Candida . All cheese samples showed contamination with varying number of yeast, total 88 yeast isolates obtained from 70 cheese samples, they were Geotrichum candidum(20.5%, Rhodotorela species(19.4%, Candida parapsilosis (18%, Candida albicans (13.6%, Candida  tropicalis (10.5%, Candida krusei (8%, Saccharomyces cerevisice (3.3% and mixed yeast (un identified at rate of (6.7%. Species of Candida formed half of the total isolates and the most prevalent isolate of Candida spp. was Candida parapsilosis .According to the results determining of  (Esterase production, Phospholipase and Hemolytic production as a virulence factors identifying Candida spp. these activities referred that all isolates of Candida spp. show one or more of these activities and that isolates of  medically important species Candida albicans were the most virulent isolates. this referred to the importance of take attention about consuming of such types of dairy products and need for applying more hygienic measures during handling, processing of milk and form of storage and/or selling of cheese.

  17. Assessment of Virulence of Raffaelea quercus-mongolicae and Raffaelea spp. Isolates by Artificial Inoculation of Quercus mongolica Logs

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    Su-Yeon Son

    2014-12-01

    Full Text Available The objective of this study was to evaluate the virulence of geographically different isolates of oak wilt pathogen, Raffaelea quercus-mongolicae and other Raffaelea species. In this study, mature trees of Quercus mongolica were inoculated with the various isolates of Raffaelea spp. and their virulence was evaluated by measuring the extent of sapwood discoloration resulting from the inoculation. The average length of discolored sapwood in a lateral direction was longest in the trees inoculated with the isolates from Korea (8.69 cm followed by R. quercivora (7.51 cm and the other Raffaelea spp. (3.35 cm. The lateral length of discolored sapwood caused by the inoculation with Korean strains varied from 4.71 to 14.90 cm indicating their differences in virulence. The area of discolored sapwood caused by the inoculation with Raffaelea spp. varied from 1.57 to 8.42 cm2 indicating their differences in virulence. Based on the length and area of the discolored sapwoods, isolated YY and wj43 appeared to have the highest virulence among all the Raffaelea isolates tested. Each of the two isolates was obtained from Gangwon Province and Jeonbuk Province, respectively.

  18. In vitro antifungal susceptibility of clinical species belonging to Aspergillus genus and Rhizopus oryzae.

    Science.gov (United States)

    Kachuei, R; Khodavaisy, S; Rezaie, S; Sharifynia, S

    2016-03-01

    Among filamentous fungal pathogens, Aspergillus spp. and zygomycetes account for highest rates of morbidity and mortality among immunocompromised patients. Recently developed antifungal drugs offer the potential to improve management and therapeutic outcomes of fungal infections. The aim of this study was to analyse the in vitro activities of voriconazole, itraconazole, amphotericin B and caspofungin against clinical isolates of Aspergillus spp. and Rhizopus oryzae. The in vitro antifungal susceptibility of 54 isolates belonging to different clinical isolates of Aspergillus spp. and R. oryzae was tested for four antifungal agents using a microdilution reference method (CLSI, M38-A2). All isolates were identified by typical colony and microscopic characteristics, and also characterized by molecular methods. Caspofungin (MEC range: 0.008-0.25 and MEC50: 0.0023μg/mL) was the most active drug in vitro against Aspergillus spp., followed by voriconazole (MIC range: 0.031-8 and MIC50: 0.5μg/mL), itraconazole (MIC range: 0.031-16 and MIC50: 0.25μg/mL), and amphotericin B (MIC range: 0.125-4 and MIC50: 0.5μg/mL), in order of decreasing activity. The caspofungin, voriconazole, and itraconazole demonstrated poor in vitro activity against R. oryzae isolates evaluated, followed by amphotericin B. This study demonstrates that caspofungin had good antifungal activity and azole agents had better activity than amphotericin B against Aspergillus species. Although, azole drugs are considered ineffective against R. oryzae. This result is just from a small scale in vitro susceptibility study and we did not take other factors into consideration. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  19. Toxin profiles of five geographical isolates of Dinophysis spp. from North and South America.

    Science.gov (United States)

    Fux, Elie; Smith, Juliette L; Tong, Mengmeng; Guzmán, Leonardo; Anderson, Donald M

    2011-02-01

    Marine dinoflagellates of the genus Dinophysis can produce toxins of the okadaic acid (OA) and pectenotoxin (PTX) groups. These lipophilic toxins accumulate in filter-feeding shellfish and cause an illness in consumers called diarrhetic shellfish poisoning (DSP). In 2008, a bloom of Dinophysis led to the closure of shellfish harvesting areas along the Texas coast, one of the first DSP-related closures in the U.S. This event resulted in a broad study of toxin production in isolates of Dinophysis spp. from U.S. waters. In the present study, we compared toxin profiles in geographical isolates of Dinophysis collected in the U.S. (Eel Pond, Woods Hole MA; Martha's Vineyard, MA; and Port Aransas Bay, Texas), and in those from Canada (Blacks Harbour, Bay of Fundy) and Chile (Reloncavi Estuary), when cultured in the laboratory under the same conditions. For each isolate, the mitochondrial cox1 gene was sequenced to assist in species identification. Strains from the northeastern U.S. and Canada were all assigned to Dinophysis acuminata, while those from Chile and Texas were most likely within the D. acuminata complex whereas precise species designation could not be made with this marker. Toxins were detected in all Dinophysis isolates and each isolate had a different profile. Toxin profiles of isolates from Eel Pond, Martha's Vineyard, and Bay of Fundy were most similar, in that they all contained OA, DTX1, and PTX2. The Eel Pond isolate also contained OA-D8 and DTX1-D7, and low levels (unconfirmed structurally) of DTX1-D8 and DTX1-D9. D. acuminata from Martha's Vineyard produced DTX1-D7, along with OA, DTX1, and PTX2, as identified in both the cells and the culture medium. D. acuminata from the Bay of Fundy produced DTX1 and PTX2, as found in both cells and culture medium, while only trace amounts of OA were detected in the medium. The Dinophysis strain from Texas only produced OA, and the one from Chile only PTX2, as confirmed in both cells and culture medium. Published

  20. Effect of gamma irradiation on growth and aflatoxin production by certain local aflatoxigenic isolates of aspergillus flavus

    International Nuclear Information System (INIS)

    Hammad, A.A.I.; Atalla, M.A.; El-Shayeb, N.M.A.; Ahmed, A.A.; Zeinat, Kamel

    1998-01-01

    The radiation D 1 0 values of two aflatoxigenic isolates of aspergillus flavus (No. 1 and No. 184) were determined. The D 1 0 -value of A. flavus isolate No.1 was found to be 0.43 and 0.50 kGy in physiological saline soulution and smoked herrings, while the D 1 0 - value of A. SFlavus isolateNo. 184 was 0.53 and -0.62 KGy in saline solution and raisins, respectively. The irradiated (0.05, 1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 KGy) and non-irradiated spores of two fungi were separately inoculated into rice-corn steep liquor and incubated at 28 degree C for 12 days. The fungal mycelial dry weight, as a function of growth, decreased with increasing gamma irradiation doses. Irradiation doses used gratly reduced aflatoxins formation. Smoked herrings and raisins were inoculated with spores of A. flavus isolated No.1 and No.184, respectively, then irradiated at 1.5, 3.0 and 4.5 KGy and stored for two months at room temperature. It was found that a gamma irradiation dose of 1.5 KGy greatly reduced the amount of aflatoxins produced, while 3.0 and 4.5 KGy completely prevented aflatoxin formation

  1. Risk factors for antibiotic resistance in Campylobacter spp. isolated from raw poultry meat in Switzerland

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    Danuser Jürg

    2003-12-01

    Full Text Available Abstract Background The world-wide increase of foodborne infections with antibiotic resistant pathogens is of growing concern and is designated by the World Health Organization as an emerging public health problem. Thermophilic Campylobacter have been recognised as a major cause of foodborne bacterial gastrointestinal human infections in Switzerland and in many other countries throughout the world. Poultry meat is the most common source for foodborne cases caused by Campylobacter. Because all classes of antibiotics recommended for treatment of human campylobacteriosis are also used in veterinary medicine, in view of food safety, the resistance status of Campylobacter isolated from poultry meat is of special interest. Methods Raw poultry meat samples were collected throughout Switzerland and Liechtenstein at retail level and examined for Campylobacter spp. One strain from each Campylobacter-positive sample was selected for susceptibility testing with the disc diffusion and the E-test method. Risk factors associated with resistance to the tested antibiotics were analysed by multiple logistic regression. Results In total, 91 Campylobacter spp. strains were isolated from 415 raw poultry meat samples. Fifty-one strains (59% were sensitive to all tested antibiotics. Nineteen strains (22% were resistant to a single, nine strains to two antibiotics, and eight strains showed at least three antibiotic resistances. Resistance was observed most frequently to ciprofloxacin (28.7%, tetracycline (12.6%, sulphonamide (11.8%, and ampicillin (10.3%. One multiple resistant strain exhibited resistance to five antibiotics including ciprofloxacin, tetracycline, and erythromycin. These are the most important antibiotics for treatment of human campylobacteriosis. A significant risk factor associated with multiple resistance in Campylobacter was foreign meat production compared to Swiss meat production (odds ratio = 5.7. Conclusion Compared to the situation in other

  2. High Milk-Clotting Activity Expressed by the Newly Isolated Paenibacillus spp. Strain BD3526

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    Feng Hang

    2016-01-01

    Full Text Available Paenibacillus spp. BD3526, a bacterium exhibiting a protein hydrolysis circle surrounded with an obvious precipitation zone on skim milk agar, was isolated from raw yak (Bos grunniens milk collected in Tibet, China. Phylogenetic analysis based on 16S rRNA and whole genome sequence comparison indicated the isolate belong to the genus Paenibacillus. The strain BD3526 demonstrated strong ability to produce protease with milk clotting activity (MCA in wheat bran broth. The protease with MCA was predominantly accumulated during the late-exponential phase of growth. The proteolytic activity (PA of the BD3526 protease was 1.33-fold higher than that of the commercial R. miehei coagulant. A maximum MCA (6470 ± 281 SU mL−1 of the strain BD3526 was reached under optimal cultivation conditions. The protease with MCA was precipitated from the cultivated supernatant of wheat bran broth with ammonium sulfate and purified by anion-exchange chromatography. The molecular weight of the protease with MCA was determined as 35 kDa by sodium dodecyl sulfate-polyacrylamide gels electrophoresis (SDS-PAGE and gelatin zymography. The cleavage site of the BD3526 protease with MCA in κ-casein was located at the Met106–Ala107 bond, as determined by mass spectrometry analysis.

  3. Antimicrobial Susceptibility of Escherichia coli Strains Isolated from Alouatta spp. Feces to Essential Oils

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    Valéria Maria Lara

    2016-01-01

    Full Text Available This study evaluated the in vitro antibacterial activity of essential oils from Lippia graveolens (Mexican oregano, Origanum vulgaris (oregano, Thymus vulgaris (thyme, Rosmarinus officinalis (rosemary, Cymbopogon nardus (citronella, Cymbopogon citratus (lemongrass, and Eucalyptus citriodora (eucalyptus against Escherichia coli (n=22 strains isolated from Alouatta spp. feces. Minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC were determined for each isolate using the broth microdilution technique. Essential oils of Mexican oregano (MIC mean = 1818 μg mL−1; MBC mean = 2618 μg mL−1, thyme (MIC mean = 2618 μg mL−1; MBC mean = 2909 μg mL−1, and oregano (MIC mean = 3418 μg mL−1; MBC mean = 4800 μg mL−1 showed the best antibacterial activity, while essential oils of eucalyptus, rosemary, citronella, and lemongrass displayed no antibacterial activity at concentrations greater than or equal to 6400 μg mL−1. Our results confirm the antimicrobial potential of some essential oils, which deserve further research.

  4. Antimicrobial Susceptibility of Escherichia coli Strains Isolated from Alouatta spp. Feces to Essential Oils

    Science.gov (United States)

    Carregaro, Adriano Bonfim; Santurio, Deise Flores; de Sá, Mariangela Facco; Santurio, Janio Moraes; Alves, Sydney Hartz

    2016-01-01

    This study evaluated the in vitro antibacterial activity of essential oils from Lippia graveolens (Mexican oregano), Origanum vulgaris (oregano), Thymus vulgaris (thyme), Rosmarinus officinalis (rosemary), Cymbopogon nardus (citronella), Cymbopogon citratus (lemongrass), and Eucalyptus citriodora (eucalyptus) against Escherichia coli (n = 22) strains isolated from Alouatta spp. feces. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for each isolate using the broth microdilution technique. Essential oils of Mexican oregano (MIC mean = 1818 μg mL−1; MBC mean = 2618 μg mL−1), thyme (MIC mean = 2618 μg mL−1; MBC mean = 2909 μg mL−1), and oregano (MIC mean = 3418 μg mL−1; MBC mean = 4800 μg mL−1) showed the best antibacterial activity, while essential oils of eucalyptus, rosemary, citronella, and lemongrass displayed no antibacterial activity at concentrations greater than or equal to 6400 μg mL−1. Our results confirm the antimicrobial potential of some essential oils, which deserve further research. PMID:27313638

  5. Characterization of expressed sequence tag-derived simple sequence repeat markers for Aspergillus flavus: emphasis on variability of isolates from the southern United States.

    Science.gov (United States)

    Wang, Xinwang; Wadl, Phillip A; Wood-Jones, Alicia; Windham, Gary; Trigiano, Robert N; Scruggs, Mary; Pilgrim, Candace; Baird, Richard

    2012-12-01

    Simple sequence repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers were designed from 362 tri-nucleotide EST-SSR sequences. Eighteen polymorphic loci were used to genotype 96 Aspergillus species isolates. The number of alleles detected per locus ranged from 2 to 24 with a mean of 8.2 alleles. Haploid diversity ranged from 0.28 to 0.91. Genetic distance matrix was used to perform principal coordinates analysis (PCA) and to generate dendrograms using unweighted pair group method with arithmetic mean (UPGMA). Two principal coordinates explained more than 75 % of the total variation among the isolates. One clade was identified for A. flavus isolates (n = 87) with the other Aspergillus species (n = 7) using PCA, but five distinct clusters were present when the others taxa were excluded from the analysis. Six groups were noted when the EST-SSR data were compared using UPGMA. However, the latter PCA or UPGMA comparison resulted in no direct associations with host species, geographical region or aflatoxin production. Furthermore, there was no direct correlation to visible morphological features such as sclerotial types. The isolates from Mississippi Delta region, which contained the largest percentage of isolates, did not show any unusual clustering except for isolates K32, K55, and 199. Further studies of these three isolates are warranted to evaluate their pathogenicity, aflatoxin production potential, additional gene sequences (e.g., RPB2), and morphological comparisons.

  6. High Proportions of Multidrug-Resistant Acinetobacter spp. Isolates in a District in Western India: A Four-Year Antibiotic Susceptibility Study of Clinical Isolates

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    Ingvild Odsbu

    2018-01-01

    Full Text Available The purpose of the study was to determine the proportions of multidrug-resistant (MDR Acinetobacter spp. isolates from the district of Nashik in Western India during the period from 2011–2014. Antibacterial susceptibility testing of isolates from inpatients and outpatients was performed using Kirby–Bauer disc diffusion method to determine inhibitory zone diameters. Proportions of non-susceptible isolates were calculated from the antibacterial susceptibility data. MDR was defined as an isolate being non-susceptible to at least one antibacterial agent in at least three antibacterial categories. The change in proportions of MDR isolates; extended-spectrum β-lactamase (ESBL-producing isolates; and non-susceptible isolates to specific antibacterial categories over calendar time was investigated by logistic regression. The proportions of MDR and ESBL-producing isolates ranged from 89.4% to 95.9% and from 87.9% to 94.0%; respectively. The proportions of non-susceptible isolates to aminoglycosides; carbapenems; antipseudomonal penicillins/β-lactamase inhibitors; cephalosporins; folate pathway inhibitors; or penicillins/β-lactamase inhibitors exceeded 77.5%. Proportions of fluoroquinolone and tetracycline non-susceptible isolates ranged from 65.3% to 83.3% and from 71.3% to 75.9%; respectively. No changes in trends were observed over time; except for a decreasing trend in fluoroquinolone non-susceptible isolates (OR = 0.75 (95% CI, 0.62–0.91. Significantly higher proportions of non-susceptible; MDR and ESBL-producing isolates were found among isolates from the respiratory system compared to isolates from all other specimen types (p < 0.05. High proportions of MDR Acinetobacter spp. isolates were observed in the period from 2011–2014. Antimicrobial stewardship programmes are needed to prevent the emergence and spread of antibiotic resistance.

  7. Isolation and identification of Bacillus spp. from compost material, compost and mushroom casing soil active against Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Stanojević Olja

    2016-01-01

    Full Text Available The isolation of bacteria was carried out from samples of straw and chicken manure, compost at various stages of the composting process and casing soil used for growing button mushrooms. A preliminary screening of 108 bacterial isolates for antagonistic activity against Trichoderma aggressivum f. europaeum showed that 23 tested isolates inhibited mycelial growth of the pathogenic fungus. Further screening with four indicator isolates of fungi revealed that all 23 bacterial isolates inhibited the growth of T. aggressivum f. europaeum, T. harzianum and T. koningii, while only 13 isolates inhibited the growth of T. atroviride. T. aggressivum f. europaeum proved to be the most sensitive, with many bacterial isolates generating a high percentage of growth inhibition. Only two bacterial isolates (B-129 and B-268 were successful in inhibiting the growth of all 4 tested pathogens. All 23 bacterial isolates were characterized as Gram-positive and catalase-positive and were subjected to molecular identification based on the partial sequence, the hypervariant region of the 16S rDNA. It was shown that the obtained bacterial strains belong to Bacillus subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus species. [Projekat Ministarstva nauke Republike Srbije, br. 31043 i br. 173026

  8. Molecular characterization of Toxocara spp. eggs isolated from public parks and playgrounds in Shiraz, Iran.

    Science.gov (United States)

    Choobineh, M; Mikaeili, F; Sadjjadi, S M; Ebrahimi, S; Iranmanesh, S

    2018-05-07

    Human toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

  9. Diversidade de fungos micorrízicos Epulorhiza spp. isolados de Epidendrum secundum (Orchidaceae Diversity of mycorrhizal fungi Epulorhiza spp. isolated from Epidendrum secundum (Orchidaceae

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    Marlon Corrêa Pereira

    2009-10-01

    Full Text Available Fungos micorrízicos rizoctonioides Epulorhiza spp. têm sido isolados de orquídeas do gênero Epidendrum e vêm sendo utilizados na germinação simbiótica das sementes de orquídeas. Epidendrum secundum é uma orquídea largamente distribuída em campos de altitude do Parque Estadual da Serra do Brigadeiro (PESB, Minas Gerais, e pouco se sabe sobre a associação micorrízica dessa espécie nesse parque. O objetivo deste trabalho foi avaliar a diversidade morfológica dos fungos micorrízicos rizoctonioides isolados de quatro populações de E. secundum em três regiões de um campo de altitude localizado na subserra Totem Deitado, PESB. Vinte e seis isolados fúngicos foram obtidos, todos pertencentes ao gênero Epulorhiza. As características morfológicas qualitativas e quantitativas avaliadas revelaram, de modo geral, baixa variabilidade entre os isolados obtidos de uma mesma população e de populações localizadas na mesma região, porém grande variabilidade foi observada entre os isolados obtidos das populações de diferentes regiões. Com base nessas características morfológicas, os isolados foram divididos em quatro grupos: o primeiro constituído pelos fungos obtidos das populações I e II da região A, o segundo pelos fungos da população III da região B, o terceiro pelo isolado M61 da população II da região A, e o quarto pelo único isolado obtido na população IV da região C. A variabilidade morfológica observada é um indicativo da diversidade dos fungos Epulorhiza spp. associados a E. secundum no PESB.Rhizoctonia-like mycorrhizal fungi Epulorhiza spp. have been isolated from orchids of the genus Epidendrum and have been used to promote the symbiotic germination of orchid seeds. Epidendrum secundum is a widely distributed orchid in campo de altitude (high elevation grassy vegetation regions of the State Park of Serra do Brigadeiro (PESB, Minas Gerais, Brazil, and little is known about the mycorrhizal relationships

  10. Aflatoxin-producing Aspergillus spp. and aflatoxin levels in stored cassava chips as affected by processing practices

    DEFF Research Database (Denmark)

    Essono, G.; Ayodele, M.; Akoa, A.

    2009-01-01

    . The levels of aflatoxin ranged between 5.2 and 14.5 ppb. The distribution of aflatoxin in positive samples depended on 8 parameters including pH, moisture content, storage duration, types of chips, level of contamination by aflatoxin-producing fungi, processing practices and storage facilities. From analysis......), followed by A. nomius (58 isolates in 15 samples), whereas A. parasiticus was rarest. A direct competitive Enzyme-linked immunosorbent assay (ELISA)-based method was implemented to quantify the content in aflatoxins. Eighteen of the samples contained some aflatoxins at detectable levels whereas 54 did not...... of variance results, only pH (p levels were...

  11. Antimicrobial Resistance of Shigella spp. isolated in the State of Pará, Brazil Resistência Antimicrobiana de Shigella spp. isoladas no Estado do Pará

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    Flávia Corrêa Bastos

    2011-10-01

    Full Text Available INTRODUCTION: Shigella spp. are Gram-negative, nonsporulating, rod-shaped bacteria that belong to the family Enterobacteriaceae and are responsible for shigellosis or bacillary dysentery, an important cause of worldwide morbidity and mortality. METHODS: We studied the antibiotic resistance profiles of 122 Shigella spp. strains (81 S. flexneri, 41 S. sonnei, 1 S. boydii isolated from patients (female and male from 0 to 80 years of age presenting diarrhea in different districts of the State of Pará, in the North of Brazil. The antibiotic resistance of the strains, isolated from human fecal samples, was determined by the diffusion disk method and by using the VITEK-2 system. RESULTS: The highest resistance rate found was the resistance rate to tetracycline (93.8%, followed by the resistance rate to chloramphenicol (63.9% and to trimethoprim/sulfamethoxazole (63.1%. Resistance to at least three drugs was more common among S. flexneri than S. sonnei (39.5% vs. 10%. Six (4.9% strains were susceptible to all the antibiotics tested. All strains were susceptible to cefotaxime, ceftazidime, ciprofloxacin, nalidixic acid and nitrofurantoin. CONCLUSIONS: High rates of multidrug resistance in Shigella spp. are a serious public health concern in Brazil. It is extremely important to continuously monitor the antimicrobial resistances of Shigella spp. for effective therapy and control measures against shigellosis.INTRODUÇÃO: Shigella spp. são bactérias Gram-negativas, não esporuladas, em forma de bastonete, pertencentes a família Enterobacteriaceae responsáveis pela shigelose ou disenteria bacilar, uma importante causa de mortalidade e morbidade mundial. MÉTODOS: Foi estudado o perfil de resistência a antimicrobianos de 122 amostras de Shigella spp. (81 S. flexneri, 41 sonnei, 1 S. boydii isoladas de pacientes (sexo feminino e masculino com faixa etária de 0 a 80 anos com distúrbios gastrointestinais em diferentes municípios no Estado do Par

  12. Antibiotic resistance in bacteria Staphylococcus spp. isolated from samples of raw sheep's milk

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    Milan Vasiľ

    2016-12-01

    Full Text Available From samples of raw sheep's milk were determined results of bacteriological examination from two herds in region of Eastern Slovakia in three years lasting study. The occurrence of Staphylococcus spp. 41.6% (124 was determined from 298 samples. The seven species of staphylococci were on a regular basis isolated: S. epidermidis (34, S. chromogenes (26, S. aureus (16. Alternately have been recorded S. warneri (16, S. schleiferi (15, S. haemolyticus (9 and S. xylosus (8. All isolated pathogens were tested by in vitro test on Mueller-Hinton agar by disc methods on resistance to 10 types of antibiotics.  Highest value of resistance was determined to Penicilin 21.0%, Neomycin 10.5% and Novobiocin 9.7%. Lower resistance was in to Oxacilin 7.2% and Amoxicilin 6.5%. Minimal resistance was founded to Cefoxitin 0.8%, Linkomycin 2.4%, Erytromycin, and Streptomycin 3.2%. Was founded total resistance (21.0% to all antibiotics in S. epidermidis (34 during the three years, S. chromogenes (26 showed resistance to 8 types of antibiotics (12.9%, S. aureus (16 to 6 antibiotics (10.5% and S. warneri (16 to 4 antibiotics (5.6%. It was confirmed that sheep's milk remains a major source of staphylococci. Bacteria in comparison with isolates from cows' raw milk, showed lower values of resistance, but were resistant to more than two antibiotics. Recorded occurrence of resistance in staphylococci may be connected with a minimum use of antibiotics in the treatment of mastitis and other diseases in sheep herds. Reported resistance to the tested antibiotics became the basis for the recommendation to use preparations to treat mastitis in sheep principally by the detection of resistance to antibiotics contained.

  13. Identification of non-Listeria spp. bacterial isolates yielding a β-D-glucosidase-positive phenotype on Agar Listeria according to Ottaviani and Agosti (ALOA).

    Science.gov (United States)

    Angelidis, Apostolos S; Kalamaki, Mary S; Georgiadou, Sofia S

    2015-01-16

    Agar Listeria according to Ottaviani and Agosti (ALOA) is the mandatory medium used for the detection and enumeration of Listeria monocytogenes in foods according to the official International Organization for Standardization (ISO) methods. On ALOA, Listeria spp. appear as bluish-green colonies due to the production of β-D-glucosidase, an enzyme that cleaves 5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside, a chromogenic substrate included in the formulation of the medium. The present work reports on bacterial isolates (n=64) from ready-to-eat soft cheeses, which are able to grow on ALOA, forming bluish-green colonies and therefore phenotypically resemble Listeria spp. All isolates were also capable of growing on the selective media PALCAM and RAPID L'mono. The isolates were characterised with biochemical tests including those specified in the ISO standards for the confirmation of Listeria spp. and identified via partial sequencing of their 16S rRNA gene. According to sequencing results the isolates represented 12 different bacterial species or species-groups belonging to seven different genera: Bacillus spp. (B. circulans, B. clausii, B. licheniformis and B. oleronius), Cellulosimicrobium spp. (C. funkei), Enterococcus spp. (E. faecalis, E. faecium/durans), Kocuria spp. (K. kristinae), Marinilactibacillus spp. (M. psychrotolerans), Rothia spp. (R. terrae) and Staphylococcus spp. (S. sciuri and S. saprophyticus subsp. saprophyticus/xylosus). Cellulosimicrobium spp. have never been previously isolated from foods. These results significantly extend the list of bacteria previously known as capable of growing on ALOA as bluish-green colonies and suggest that there may be room for further improvement in the medium's inhibitory properties towards non-Listeria spp., Gram-positive bacteria present in foods. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Isolation and risk assessment of Geotrichum spp. in the white shrimp (Litopenaeus vannamei Boone, 1931 from culture ponds

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    José Luis Ochoa

    2015-09-01

    Full Text Available The present study was done in order to identify the fungus invading some of the supralittoral ponds used for shrimp aquaculture in the CIBNOR facilities in La Paz, Baja California Sur (BCS, México during the summer season. From the walls and bottoms of the ponds, two strains of Geotrichum spp. were isolated and morphologically identified. Fungal adhesion towards hemocytes and primary cultures of various white shrimp (Litopeneaus vannamei tissues (gill, tegument, and gut was analyzed to determine infectivity. Extracellular protease, lipase, and amylase activity were evaluated as virulence factors. Survival of shrimp post-larvae (PL8 exposed to fungal culture supernatant or to their filaments was also investigated. The results showed that shrimp tegument cells and hemocytes were very susceptible to Geotrichum spp. invasion, and that this fungus provokes great mortality of post-larvae. Hence, Geotrichum spp. could be considered an opportunistic pathogen that might represent a serious health risk to shrimp in culture.

  15. Biofilm Filtrates of Pseudomonas aeruginosa Strains Isolated from Cystic Fibrosis Patients Inhibit Preformed Aspergillus fumigatus Biofilms via Apoptosis.

    Science.gov (United States)

    Shirazi, Fazal; Ferreira, Jose A G; Stevens, David A; Clemons, Karl V; Kontoyiannis, Dimitrios P

    2016-01-01

    Pseudomonas aeruginosa (Pa) and Aspergillus fumigatus (Af) colonize cystic fibrosis (CF) patient airways. Pa culture filtrates inhibit Af biofilms, and Pa non-CF, mucoid (Muc-CF) and nonmucoid CF (NMuc-CF) isolates form an ascending inhibitory hierarchy. We hypothesized this activity is mediated through apoptosis induction. One Af and three Pa (non-CF, Muc-CF, NMuc-CF) reference isolates were studied. Af biofilm was formed in 96 well plates for 16 h ± Pa biofilm filtrates. After 24 h, apoptosis was characterized by viability dye DiBAc, reactive oxygen species (ROS) generation, mitochondrial membrane depolarization, DNA fragmentation and metacaspase activity. Muc-CF and NMuc-CF filtrates inhibited and damaged Af biofilm (pbiofilms (3.7- fold) compared to treatment with filtrates from Muc-CF- (2.5- fold) or non-CF Pa (1.7- fold). Depolarization of mitochondrial potential was greater upon exposure to NMuc-CF (2.4-fold) compared to Muc-CF (1.8-fold) or non-CF (1.25-fold) (pbiofilm, compared to control, mediated by metacaspase activation. In conclusion, filtrates from CF-Pa isolates were more inhibitory against Af biofilms than from non-CF. The apoptotic effect involves mitochondrial membrane damage associated with metacaspase activation.

  16. Distribution of genes encoding virulence factors and molecular analysis of Shigella spp. isolated from patients with diarrhea in Kerman, Iran.

    Science.gov (United States)

    Hosseini Nave, Hossein; Mansouri, Shahla; Emaneini, Mohammad; Moradi, Mohammad

    2016-03-01

    Shigella is one of the important causes of diarrhea worldwide. Shigella has several virulence factors contributing in colonization and invasion of epithelial cells and eventually death of host cells. The present study was performed in order to investigate the distribution of virulence factors genes in Shigella spp. isolated from patients with acute diarrhea in Kerman, Iran as well as the genetic relationship of these isolates. A total of 56 isolates including 31 S. flexneri, 18 S. sonnei and 7 S. boydii were evaluated by polymerase chain reaction (PCR) for the presence of 11 virulence genes (ipaH, ial, set1A, set1B, sen, virF, invE, sat, sigA, pic and sepA). Then, the clonal relationship of these strains was analyzed by multilocus variable-number tandem repeat analysis (MLVA) method. All isolates were positive for ipaH gene. The other genes include ial, invE and virF were found in 80.4%, 60.7% and 67.9% of the isolates, respectively. Both set1A and set1B were detected in 32.3% of S. flexneri isolates, whereas 66.1% of the isolates belonging to different serogroup carried sen gene. The sat gene was present in all S. flexneri isolates, but not in the S. sonnei and S. boydii isolates. The result showed, 30.4% of isolates were simultaneously positive and the rest of the isolates were negative for sepA and pic genes. The Shigella isolates were divided into 29 MLVA types. This study, for the first time, investigated distribution of 11 virulence genes in Shigella spp. Our results revealed heterogeneity of virulence genes in different Shigella serogroups. Furthermore, the strains belonging to the same species had little diversity. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Isolation and Molecular Characterization of Potential Plant Growth Promoting Bacillus cereus GGBSTD1 and Pseudomonas spp. GGBSTD3 from Vermisources

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    Balayogan Sivasankari

    2014-01-01

    Full Text Available Vermicompost was prepared from leaf materials of Gliricidia sepium + Cassia auriculata + Leucaena leucocephala with cow dung (1 : 1 : 2 using Eudrilus eugeniae (Kinberg and Eisenia fetida for 60 days. Nineteen bacterial strains which have the capability to fix nitrogen, solubilize inorganic phosphate, and produce phytohormones were isolated from vermicompost, vermisources, and earthworm (fore, mid, and hind guts and tested for plant growth studies. Among the bacterial strains only five strains had both activities; among the five Bacillus spp. showed more nitrogen fixing activity and Pseudomonas spp. showed more phosphate solubilizing activity. Hence these bacterial strains were selected for further molecular analysis and identified Bacillus cereus GGBSTD1 and Pseudomonas spp. GGBSTD3. Plant growth studies use these two organisms separately and as consortium (Bacillus cereus + Pseudomonas spp. in (1 : 1 ratio at different concentrations using Vigna unguiculata (L. Walp. at different day intervals. The germination percent, shoot length, root length, leaf area, chlorophyll a content of the leaves, chlorophyll b content of the leaves, total chlorophyll content of the leaves, fresh weight of the whole plant, and dry weight of the whole plant were significantly enhanced by the consortium (Bacillus cereus + Pseudomonas spp. of two organisms at 5 mL concentrations on the 15th day compared to others.

  18. Isolation, antibiogram and pathogenicity of Salmonella spp. Recovered from slaughtered food animals in Nagpur region of Central India

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    D. G. Kalambhe

    2016-02-01

    Full Text Available Aim: To determine the prevalence, antibiogram and pathogenicity of Salmonella spp. in the common food animals slaughtered for consumption purpose at government approved slaughter houses located in and around Nagpur region during a period of 2010-2012. Materials and Methods: A total of 400 samples comprising 50 each of blood and meat from each slaughtered male cattle, buffaloes, pigs and goats were collected. Isolation was done by pre-enrichment in buffered peptone water and enrichment in Rappaport-Vassiliadis broth with subsequent selective plating onto xylose lysine deoxycholate agar. Presumptive Salmonella colonies were biochemically confirmed and analyzed for pathogenicity by hemolysin production and Congo red dye binding assay (CRDA. An antibiotic sensitivity test was performed to assess the antibiotic resistance pattern of the isolates. Results: A total of 10 isolates of Salmonella spp. from meat (3 from cattle, 1 from buffaloes and 6 from pigs with an overall prevalence of 5% among food animals was recorded. No isolation was reported from any blood samples. Pathogenicity assays revealed 100% and 80% positivity for CRDA and hemolytic activity, respectively. Antimicrobial sensitivity test showed multi-drug resistance. The overall resistance of 50% was noted for trimethoprim followed by ampicillin (20%. A maximum sensitivity (80% was reported to gentamycin followed by 40% each to ampicillin and trimethoprim, 30% to amikacin and 10% to kanamycin. Conclusion: The presence of multidrug resistant and potentially pathogenic Salmonella spp. in slaughtered food animals in Nagpur region can be a matter of concern for public health.

  19. Mycogenic synthesis of silver nanoparticles by the Japanese environmental isolate Aspergillus tamarii

    Energy Technology Data Exchange (ETDEWEB)

    Rajesh Kumar, Ramasamy; Poornima Priyadharsani, Krishnamurthy; Thamaraiselvi, Kaliannan, E-mail: kthamaraiselvi@hotmail.com [Bharathidasan University, Laboratory of Molecular Microbial Bioremediation and Nanobiotechnology, Department of Environmental Biotechnology (India)

    2012-05-15

    In this study, an environmental friendly process for the synthesis of silver nanoparticles (AgNPs) using a fungus Aspergillus tamarii has been investigated. The process of silver ion reduction by the fungal extracellular filtrate was spontaneous which lead to the development of an easy process for synthesis of silver nanoparticles. The AgNPs formed were characterized using UV-Visible spectrum, FTIR, and SEM. The results revealed that silver ions reduction by the fungal extracellular filtrate started at 420 nm after 0.5 h of incubation time. The FTIR peaks were observed at 1393, 1820, 2727, and 3545 cm{sup -1}. The SEM result showed the distribution of spherical AgNPs ranging from 25 to 50 nm.

  20. Isolation and Biological Evaluation of Two Bioactive Metabolites from Aspergillus gorakhpurensis

    Directory of Open Access Journals (Sweden)

    Venkateswarlu Yenamandra

    2009-06-01

    Full Text Available Fungi are known to produce a vast array of secondary metabolites that are gaining importance for their biotechnological applications. Screening of Aspergillus gorakhpurensis for the production of bioactive secondary metabolites results in the production of 4-(N-methyl-N-phenyl amino butan-2-one and itaconic acid. The structure of the known compounds was established by 1H-, 13C-NMR and Mass spectral data. Biological evaluation of the two compounds against test microorganisms showed strong inhibitory activity of 4-(N-methyl-N-phenyl amino butan-2-one towards bacteria and fungi. Only 4-( N -methyl-N- phenyl amino-butan-2-one showed a marked significant activity (LD 50 = 330.69 m g/mL in Spodoptera litura larvicidal bioassay.

  1. Antifungal activity of linalool in cases of Candida spp. isolated from individuals with oral candidiasis

    Directory of Open Access Journals (Sweden)

    I. J. Dias

    2017-09-01

    Full Text Available Abstract This study analyzed the antifungal activity of phytoconstituents from linalool on Candida spp. strains, in vitro, isolated from patients with clinical diagnoses of oral candidiasis associated with the use of a dental prosthesis. Biological samples were collected from 12 patients using complete dentures or removable partial dentures and who presented mucous with diffuse erythematous or stippled features, indicating a clinical diagnosis of candidiasis. To identify fungal colonies of the genus Candida, samples were plated onto CHROMagar Candida®. The antifungal activity of linalool, a monoterpene unsaturated constituent of basil oil, was performed using the broth microdilution technique. Then, the minimum inhibitory concentration (MIC, the two subsequent stronger concentrations and the positive controls were subcultured on Sabouraud Dextrose Agar plates to determine the minimum fungicidal concentration (MFC. The experiments were performed in triplicate and nystatin was used as a positive control in all tests. Diagnoses of oral candidiasis were verified in eight patients (66.6% and the most prevalent fungal species was Candida albicans (37.5%, followed by Candida krusei (25.0%; and Candida tropicalis (4.2%. The best antifungal activity of linalool was observed on Candida tropicalis (MIC = 500 mg/mL, followed by Candida albicans (MIC = 1.000 mg/mL, and Candida krusei (MIC = 2.000 mg/mL.Under the study conditions and based on the results obtained, it can be concluded that the Candida strains tested were susceptible to linalool.

  2. Isolation and Characterization of an α-Glucosidase Inhibitor from Musa spp. (Baxijiao Flowers

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    Zhanwu Sheng

    2014-07-01

    Full Text Available The use of α-glucosidase inhibitors is considered to be an effective strategy in the treatment of diabetes. Using a bioassay-guided fractionation technique, five Bacillus stearothermophilus α-glucosidase inhibitors were isolated from the flowers of Musa spp. (Baxijiao. Using NMR spectroscopy analysis they were identified as vanillic acid (1, ferulic acid (2, β-sitosterol (3, daucosterol (4 and 9-(4′-hydroxyphenyl-2-methoxyphenalen-1-one (5. The half maximal inhibitory concentration (IC50 values of compounds 1–5 were 2004.58, 1258.35, 283.67, 247.35 and 3.86 mg/L, respectively. Compared to a known α-glucosidase inhibitor (acarbose, IC50 = 999.31 mg/L, compounds 3, 4 and 5 showed a strong α-glucosidase inhibitory effect. A Lineweaver-Burk plot indicated that compound 5 is a mixed-competitive inhibitor, while compounds 3 and 4 are competitive inhibitors. The inhibition constants (Ki of compounds 3, 4 and 5 were 20.09, 2.34 and 4.40 mg/L, respectively. Taken together, these data show that the compounds 3, 4 and 5 are potent α-glucosidase inhibitors.

  3. Identification, antimicrobial resistance and genotypic characterization of Enterococcus spp. isolated in Porto Alegre, Brazil

    Science.gov (United States)

    Bender, Eduardo André; de Freitas, Ana Lúcia Peixoto; Reiter, Keli Cristine; Lutz, Larissa; Barth, Afonso Luís

    2009-01-01

    In the past two decades the members of the genus Enterococcus have emerged as important nosocomial pathogens worldwide. In the present study, we evaluated the antimicrobial resistance and genotypic characteristics of 203 Enterococcus spp. recovered from different clinical sources from two hospitals in Porto Alegre, Rio Grande do Sul, Brazil. The species were identified by conventional biochemical tests and by an automated system. The genetic diversity of E. faecalis presenting high-level aminoglycoside resistance (HLAR) was assessed by pulsed-field gel electrophoresis of chromosomal DNA after SmaI digestion. The E. faecalis was the most frequent specie (93.6%), followed by E. faecium (4.4%). The antimicrobial resistance profile was: 2.5% to ampicillin, 0.5% to vancomycin, 0.5% teicoplanin, 33% to chloramphenicol, 2% to nitrofurantoin, 66.1% to erythromycin, 66.5% to tetracycline, 24.6% to rifampicin, 30% to ciprofloxacin and 87.2% to quinupristin-dalfopristin. A total of 10.3% of the isolates proved to be HLAR to both gentamicin and streptomycin (HLR-ST/GE), with 23.6% resistant only to gentamicin (HLR-GE) and 37.4% only to streptomycin (HLR-ST). One predominant clonal group was found among E. faecalis HLR-GE/ST. The prevalence of resistance among beta-lactam antibiotics and glycopeptides was very low. However, in this study there was an increased number of HLR Enterococcus which may be spreading intra and inter-hospital. PMID:24031416

  4. Antimicrobial susceptibility of Brachyspira spp. isolated from commercial laying hens and free-living wild mallards (Anas platyrhynchos).

    Science.gov (United States)

    Jansson, Désirée S; Pringle, Märit

    2011-08-01

    In vitro antimicrobial susceptibility to tylosin, valnemulin, tiamulin, doxycycline, lincomycin and ampicillin was investigated by broth dilution in 48 Brachyspira spp. isolates from commercial laying hens (n=30) and free-living wild mallards (Anas platyrhynchos) (n=18). Presumed pathogens (Brachyspira alvinipulli, Brachyspira intermedia, Brachyspira pilosicoli), commensals (Brachyspira murdochii, Brachyspira innocens, "Brachyspira pulli"), and isolates of undetermined species affiliation were included. The laying hens had not been exposed to therapeutic levels of antimicrobials for at least 50 weeks before sampling, and low levels of environmental antimicrobial exposure were presumed in mallards. No isolates with decreased susceptibility to tylosin, valnemulin, tiamulin or doxycycline were found. Decreased susceptibility to lincomycin (minimum inhibitory concentration 16 µg/ml) was detected in two isolates (Brachyspira sp.) from laying hens. Five isolates showed decreased susceptibility to ampicillin (minimum inhibitory concentration 16 to >32 µg/ml), including two "B. pulli" and one B. alvinipulli from laying hens, and isolates of B. pilosicoli and "B. pulli" from mallards. Decreased susceptibility to ampicillin was associated with β-lactamase activity in four isolates. A new variant of a class D β-lactamase gene designated bla (oxa-192) was identified in a B. pilosicoli isolate of mallard origin. This is the first time the genetic basis for antimicrobial resistance is described in Brachyspira spp. from a free-living wild bird. Isolates displaying decreased susceptibility to ampicillin were accompanied by fully susceptible isolates of the same species or other genotypes within three laying hen flocks. This underlines the need for performing antimicrobial susceptibility tests on single clones/genotypes, and to analyse multiple isolates from the same flock.

  5. Improved protocol for isolation of Campylobacter spp. from retail broiler meat and use of pulsed field gel electrophoresis for the typing of isolates.

    Science.gov (United States)

    Oyarzabal, Omar A; Williams, Aretha; Zhou, Ping; Samadpour, Mansour

    2013-10-01

    To improve the detection of Campylobacter spp. in retail broiler meat, a reference method (R subsamples) based on the enrichment of 25 g of meat in Bolton broth at 42°C under microaerobiosis was compared with an alternative method (A subsamples) consisting in the rinsing of meat samples for 30s in buffered peptone water with antimicrobials with incubation at 42°C under aerobiosis. One piece of meat (breasts, tenderloins and thighs) was rinse in experiment 1 (A1) and two pieces in experiment 2 (A2). Campylobacter spp. were isolated on agar plates and identified by PCR. Retail samples in Alabama had less prevalence (P ≤ 0.05) than samples in the state of Washington. The percentage of positive was higher (P ≤ 0.05) in A than in R subsamples and rinsing two pieces of meat yielded the highest percentage of positive subsamples. R subsamples showed variations in the prevalence by product. However, A subsamples had similar prevalence of positives among products compare to the result from reference method. More Campylobacter coli isolates were collected in A2 subsamples. Pulse field gel electrophoresis (PFGE) was used as subtyping method to study the genome similarity among the isolates from all methods. A larger diversity of isolates were detected by PFGE in A2 subsamples. Denaturing gradient gel electrophoresis analysis suggested that the initial bacterial populations of the meat samples impact the final bacterial profile after enrichment. Rinsing broiler meats was less time consuming, required less sample preparation and was more sensitive than the reference method for the isolation of naturally occurring Campylobacter spp. This new method could help with epidemiological and intervention studies to control Campylobacter spp. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Fungi in spices and mycotoxigenic potential of some Aspergilli isolated.

    Science.gov (United States)

    Garcia, Marcelo Valle; Parussolo, Gilson; Moro, Camila Brombilla; Bernardi, Angélica Olivier; Copetti, Marina Venturini

    2018-08-01

    The aim of this study was to identify fungal species present in 200 samples of rosemary, fennel, cinnamon, clove, pepperoni, black and white pepper and oregano and evaluate the mycotoxigenic potential of the some Aspergilli isolated. Clove, black and white peppers were analyzed by direct plating. For rosemary, cinnamon, fennel, pepperoni pepper and oregano samples were used spread plate. Mycotoxigenic capacity was verified by the agar plug method. With the exception of clove, all the spices showed high fungal contamination, especially by Aspergillus sp., Penicillium sp. and Cladosporium sp. Frequency of toxigenic Aspergillus spp. was intense in white and black peppers, with presence of Aspergillus flavus (up to 32%), Aspergillus nomius (up to 12%), Aspergillus parasiticus (up to 4%), Aspergillus niger complex (up to 52%), Aspergillus ochraceus (up 12%) and Aspergillus carbonarius (up to 4%). 14,2% of A. flavus isolated from black pepper were aflatoxins producers. In the white pepper, 66.7% of A. flavus isolates and 100% of A. nomius were aflatoxigenic. Oregano showed the highest number of A. niger complex isolates (49), however, only 2.04% produced ochratoxin A. This study showed a huge fungal presence in spices, which could compromise the sensorial quality of these products and represent a hazard for consumers. Copyright © 2018. Published by Elsevier Ltd.

  7. Discrimination of Aspergillus lentulus from Aspergillus fumigatus by Raman spectroscopy and MALDI-TOF MS.

    Science.gov (United States)

    Verwer, P E B; van Leeuwen, W B; Girard, V; Monnin, V; van Belkum, A; Staab, J F; Verbrugh, H A; Bakker-Woudenberg, I A J M; van de Sande, W W J

    2014-02-01

    In 2005, a new sibling species of Aspergillus fumigatus was discovered: Aspergillus lentulus. Both species can cause invasive fungal disease in immune-compromised patients. The species are morphologically very similar. Current techniques for identification are PCR-based or morphology-based. These techniques are labour-intense and not sufficiently discriminatory. Since A. lentulus is less susceptible to several antifungal agents, it is important to correctly identify the causative infectious agent in order to optimize antifungal therapy. In this study we determined whether Raman spectroscopy and/or MALDI-TOF MS were able to differentiate between A. lentulus and A. fumigatus. For 16 isolates of A. lentulus and 16 isolates of A. fumigatus, Raman spectra and peptide profiles were obtained using the Spectracell and MALDI-TOF MS (VITEK MS RUO, bioMérieux) respectively. In order to obtain reliable Raman spectra for A. fumigatus and A. lentulus, the culture medium needed to be adjusted to obtain colourless conidia. Only Raman spectra obtained from colourless conidia were reproducible and correctly identified 25 out of 32 (78 %) of the Aspergillus strains. For VITEK MS RUO, no medium adjustments were necessary. Pigmented conidia resulted in reproducible peptide profiles as well in this case. VITEK MS RUO correctly identified 100 % of the Aspergillus isolates, within a timeframe of approximately 54 h including culture. Of the two techniques studied here, VITEK MS RUO was superior to Raman spectroscopy in the discrimination of A. lentulus from A. fumigatus. VITEK MS RUO seems to be a successful technique in the daily identification of Aspergillus spp. within a limited timeframe.

  8. Antifungal susceptibility testing of Aspergillus species complex in the Clinical Laboratory: how to do it, when to do it, and how to interpret it

    Directory of Open Access Journals (Sweden)

    Esther Manso

    2014-12-01

    Full Text Available The emergence of drug resistance in fungal pathogens has a profound impact on human health given limited number of antifungal drugs. Antifungal resistance in Aspergillus spp. infection can be encountered in the antifungal drug-exposed patient due to selection of intrinsically resistant species or isolates with acquired resistance belonging to species that are normally susceptible. Resistance to triazoles is not common in Aspergillus spp., however, triazole resistance in A. fumigatus appears to be increasing in several European countries in recent years and can be clinically relevant. The Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing have developed breakpoints and epidemiological cutoff values that are now established for Aspergillus spp. Clinical microbiology laboratories will be employed commercial susceptibility assays, rather than reference broth microdilution methods and comparative studies are particularly important.

  9. Species distribution, virulence factors, and antimicrobial resistance of Acinetobacter spp. isolates from dogs and cats: a preliminary study.

    Science.gov (United States)

    Kimura, Yui; Harada, Kazuki; Shimizu, Takae; Sato, Tomomi; Kajino, Akari; Usui, Masaru; Tamura, Yutaka; Tsuyuki, Yuzo; Miyamoto, Tadashi; Ohki, Asami; Watarai, Masahisa

    2018-05-12

    We investigated the prevalence of virulence factors and antimicrobial resistance among 67 Acinetobacter spp. isolates, consisting of 21 Acinetobacter baumannii and 46 non-baumannii Acinetobacter from companion animals. The PCR analysis showed that the most prevalent virulence gene was afa/draBC (29.9%), followed by papC (22.4%) and cvaC (20.9%). Antimicrobial susceptibility testing revealed that resistance to gentamicin (14.9%) and ciprofloxacin (11.9%) was relatively prevalent. Five gentamicin- and/or ciprofloxacin-resistant A. baumannii strains were assigned to ST25, ST149, ST164, ST203, and ST1198. All ciprofloxacin-resistant isolates harbored point mutations in gyrA and/or parC. This is the first preliminary monitoring of animal-origin Acinetobacter spp. in Japan. © 2018 The Societies and John Wiley & Sons Australia, Ltd.

  10. Genetic and phenotypic diversity of naturally isolated wild strains of Aspergillus niger with hyper glucose oxidase production

    Directory of Open Access Journals (Sweden)

    MAHMOUD EL-HARIRI

    2015-12-01

    Full Text Available Glucose oxidase (GOx is the basic stone for many of biological industry worldwide. The improvement of GOx production basically depends on selection of hyper producer strain of Aspergillus niger. Selective isolation and screening for natural hyper producer strains of A. niger and sequence analysis of the GOD gene, which is responsible for production of the enzyme, are very effective approaches to investigate the naturally modified strains of A. niger with hyper productive capacity of GOx enzyme. The aims of the current study were selective isolation of naturally hyper GOx producing strains of A. niger and evaluation of their GOx activities under optimized parameters in the laboratory. Five wild Egyptian isolates of A. niger were screened for GOx and catalase activity using two types of modified basal liquid media. The GOx activity was evaluated by high throughout liquid phase system. The isolates showed a variable activity for GOx production ranged from 0 to 28.7 U.ml-1. One isolate coded Strain 7 was negative GOx producer on Vogel's broth medium in comparison to other isolates, while its GOx activity on Cazpek Dox was considered as positive (7.28 U.ml-1. It was concluded that GOx production is affected by three controllable factors – the basal media components, time of incubation, and the strain with its adaption to the media components‎. Also, the catalase activity was tested and it was produced with a different degree of variability, which may be reflected on GOx stability. GOD genes of these wild variant of A. niger were cloned and sequenced to determine intraspecies diversity of GOD between the wild variants. The comparison of isolated wild variants to other reference hyper GOx producer strains of A. niger was performed to determine if the GOD sequence analysis of these strains can be distinguished based on their GOx activity. This is the first report for isolation and detection of naturally A. niger hyper GOx-producer strains with

  11. PATHOGENICITY OF FUSARIUM SPP. ISOLATED FROM WEEDS AND PLANT DEBRIS IN EASTERN CROATIA TO WHEAT AND MAIZE

    Directory of Open Access Journals (Sweden)

    Jelena Ilić

    2012-12-01

    Full Text Available Pathogenicity of thirty isolates representing 14 Fusarium species isolated from weeds and plant debris in eastern Croatia was investigated in the laboratory. Pathogenicity tests were performed on wheat and maize seedlings. The most pathogenic Fusarium spp. was F. graminearum isolated from Amaranthus retroflexus, Abutilon theophrasti and Chenopodium album. There was a noticeable inter- and intraspecies variability in pathogenicity towards wheat and maize. Isolates of F. solani from Sonchus arvensis and F. verticillioides from C. album were highly pathogenic to wheat seedlings and apathogenic to maize seedlings. Isolates of F. venenatum were very pathogenic to wheat and maize being the first report about pathogenicity of this species. This experiment proves that weeds and plant debris can serve as alternate hosts and source of inoculum of plant pathogens.

  12. Aspergillus section Fumigati typing by PCR-restriction fragment polymorphism.

    Science.gov (United States)

    Staab, Janet F; Balajee, S Arunmozhi; Marr, Kieren A

    2009-07-01

    Recent studies have shown that there are multiple clinically important members of the Aspergillus section Fumigati that are difficult to distinguish on the basis of morphological features (e.g., Aspergillus fumigatus, A. lentulus, and Neosartorya udagawae). Identification of these organisms may be clinically important, as some species vary in their susceptibilities to antifungal agents. In a prior study, we utilized multilocus sequence typing to describe A. lentulus as a species distinct from A. fumigatus. The sequence data show that the gene encoding beta-tubulin, benA, has high interspecies variability at intronic regions but is conserved among isolates of the same species. These data were used to develop a PCR-restriction fragment length polymorphism (PCR-RFLP) method that rapidly and accurately distinguishes A. fumigatus, A. lentulus, and N. udagawae, three major species within the section Fumigati that have previously been implicated in disease. Digestion of the benA amplicon with BccI generated unique banding patterns; the results were validated by screening a collection of clinical strains and by in silico analysis of the benA sequences of Aspergillus spp. deposited in the GenBank database. PCR-RFLP of benA is a simple method for the identification of clinically important, similar morphotypes of Aspergillus spp. within the section Fumigati.

  13. Aspergillus Section Fumigati Typing by PCR-Restriction Fragment Polymorphism▿

    Science.gov (United States)

    Staab, Janet F.; Balajee, S. Arunmozhi; Marr, Kieren A.

    2009-01-01

    Recent studies have shown that there are multiple clinically important members of the Aspergillus section Fumigati that are difficult to distinguish on the basis of morphological features (e.g., Aspergillus fumigatus, A. lentulus, and Neosartorya udagawae). Identification of these organisms may be clinically important, as some species vary in their susceptibilities to antifungal agents. In a prior study, we utilized multilocus sequence typing to describe A. lentulus as a species distinct from A. fumigatus. The sequence data show that the gene encoding β-tubulin, benA, has high interspecies variability at intronic regions but is conserved among isolates of the same species. These data were used to develop a PCR-restriction fragment length polymorphism (PCR-RFLP) method that rapidly and accurately distinguishes A. fumigatus, A. lentulus, and N. udagawae, three major species within the section Fumigati that have previously been implicated in disease. Digestion of the benA amplicon with BccI generated unique banding patterns; the results were validated by screening a collection of clinical strains and by in silico analysis of the benA sequences of Aspergillus spp. deposited in the GenBank database. PCR-RFLP of benA is a simple method for the identification of clinically important, similar morphotypes of Aspergillus spp. within the section Fumigati. PMID:19403766

  14. ANTIBACTERIAL POTENTIAL OF NATIVE PLANTS FROM THE CAATINGA BIOME AGAINST Staphylococcus spp. ISOLATES FROM SMALL RUMINANTS WITH MASTITIS

    OpenAIRE

    PEIXOTO, RODOLFO DE MORAES; SILVA, WELLINGTON ERASMO LIMA E; ALMEIDA, JACKSON ROBERTO GUEDES SILVA; BRANCO, ALEXSANDRO; COSTA, MATEUS MATIUZZI DA

    2016-01-01

    ABSTRACT The aim of the present study is to assess the antibacterial potential of plants from the Caatinga biome of the semi-arid region of Pernambuco, against Staphylococcus spp. isolates from cases of subclinical mastitis in small ruminants, such as goats and ewes. Ethanolic extracts of the following plants from the Caatinga biome were used: Encholirium spectabile Mart., Bromelia laciniosa Mart., Neoglaziovia variegata Mez., Amburana cearensis (Fr. Allem.) A. C. Smith, Hymenaea martiana Hay...

  15. Isolation of Arcobacter spp from the milk of dairy cows in Brazil Isolamento de Arcobacter spp do leite de vacas leiteiras no Brasil

    Directory of Open Access Journals (Sweden)

    Celso Pianta

    2007-02-01

    Full Text Available Bacteriologic examinations were performed on 188 milk samples collected from cows from 11 farms for diagnosis of mastitis in three cities of Rio Grande do Sul, Brazil. Among the common causes of mastitis, the most frequent isolates were Staphylococcus aureus, followed by Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae and Streptococcus agalactiae. Bacteriologic examination of 32 milk samples from one farm didn't show bacteria known as common etiologic agent of mastitis. Six samples of Arcobacter spp typed by genotypic tests as Arcobacter cryaerophilus (five strains and Arcobacter butzleri (one strain were isolated from cows' milk of that farm. It is reported the isolation of Arcobacter species from the milk of cows in absence of clinical signs of mastitis. This is the first report of the detection of the microorganisms in the milk of dairy cows in Brazil. No previous reports are known from other countries.Foram realizados exames bacteriológicos em 188 amostras de leite colhidas de vacas de 11 propriedades leiteiras para diagnóstico de mastite, em três municípios no Rio Grande do Sul, Brasil. Entre as causas comuns de mastite, os isolados mais freqüentes foram Staphylococcus aureus, seguido de Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae e Streptococcus agalactiae. O exame bacteriológico realizado em 32 amostras de leite de vacas de uma propriedade não demonstrou a presença de bactérias conhecidas como causadoras de mastite. Foram isoladas do leite de vacas desta propriedade seis amostras de Arcobacter spp, classificadas por testes moleculares como Arcobacter cryaerophilus (cinco amostras e Arcobacter butzleri (uma amostra. É relatado o isolamento de espécies de Arcobacter do leite de vacas na ausência de sinais clínicos de mastite. Este é o primeiro relato da detecção dos microorganismos no leite de vacas leiteiras no Brasil.

  16. Microbial Observatory (ISS-MO): Draft Genome Sequence of two Aspergillus fumigatus Strains Isolated from the International Space Station

    Data.gov (United States)

    National Aeronautics and Space Administration — Aspergillus fumigatus is a saprophytic filamentous fungus that is ubiquitous outdoors (soil decaying vegetation) and indoors (hospitals simulated closed habitats...

  17. [Sensitivity profile of Staphylococcus spp. and Streptococcus spp. isolated from toys used in a teaching hospital playroom].

    Science.gov (United States)

    Boretti, Vanessa Stolf; Corrêa, Renata Nunes; dos Santos, Silvana Soléo Ferreira; Leão, Mariella Vieira Pereira; Gonçalves e Silva, Célia Regina

    2014-09-01

    To evaluate the presence of microorganisms of the genus Staphylococcus and Streptococcus on toys in the playroom of a teaching hospital, as well to as analyze the antimicrobial from the isolated strains. Samples were collected from 60 toys, using wet swabs, soon after being used by the children. The samples were inoculated in enriched and selective agar for isolation and later identification of the microorganisms. Antibiogram testing was performed by agar diffusion technique. The genus Staphylococcus was present in 87.0% (52/60) of the toys. Seventythree strains were isolated, with 29.0% (21/73) coagulase-positive and 71.0% (52/73) coagulase-negative. Among the coagulase-negative strains, 90.4% were resistant to penicillin, 65.4% to oxacillin, 28.8% to clarithromycin, 61.5% to clindamycin, and none to vancomycin. Among the coagulase-positive strains, 76.2% were resistant to penicillin, 23.8% to oxacillin, 23.8% to clarithromycin, 47.6% to clindamycin, and none to vancomycin. The genus Streptococcus was not detected in any of the evaluated toys. Toys can be contaminated with potentially pathogenic bacteria with antimicrobial resistance, representing a possible source of nosocomial infection for patients who are already debilitated. Copyright © 2014 Sociedade de Pediatria de São Paulo. Publicado por Elsevier Editora Ltda. All rights reserved.

  18. Sensitivity profile of Staphylococcus spp. and Streptococcus spp. isolated from toys used in a teaching hospital playroom☆

    Science.gov (United States)

    Boretti, Vanessa Stolf; Corrêa, Renata Nunes; dos Santos, Silvana Soléo Ferreira; Leão, Mariella Vieira Pereira; Silva, Célia Regina Gonçalves e

    2014-01-01

    Objective: To evaluate the presence of microorganisms of the genus Staphylococcus and Streptococcus on toys in the playroom of a teaching hospital, as well to as analyze the antimicrobial resistance from isolated strains. Methods: Samples were collected from 60 toys, using wet swabs, soon after being used by the children. The samples were inoculated in enriched and selective agar for isolation and later identification of the microorganisms. Antibiogram testing was performed by agar diffusion technique. Results: The genus Staphylococcus was present in 87.0% (52/60) of the toys. Seventy-three strains were isolated, with 29.0% (21/73) coagulase-positive and 71.0% (52/73) coagulasenegative. Among the coagulase-negative strains, 90.4% were resistant to penicillin, 65.4% to oxacillin, 28.8% to clarithromycin, 61.5% to clindamycin, and none to vancomycin. Among the coagulase-positive strains, 76.2% were resistant to penicillin, 23.8% to oxacillin, 23.8% to clarithromycin, 47.6% to clindamycin, and none to vancomycin. The genus Streptococcus was not detected in any of the evaluated toys. Conclusions: Toys can be contaminated with potentially pathogenic bacteria with antimicrobial resistance, representing a possible source of nosocomial infection for patients who are already debilitated. PMID:25479842

  19. Determination of antifungal susceptibility patterns among the environmental isolates of Aspergillus fumigatus in Iran

    Directory of Open Access Journals (Sweden)

    Faezeh Mohammadi

    2016-01-01

    Conclusion: Our findings demonstrated that there was not azole-resistant among environmental isolates of A. fumigatus. Medical triazoles compounds have structural similarity with triazole fungicide compounds in agriculture, therefore, resistance development through exposure to triazole fungicide compounds in the environment is important but it sounds there is not a serious health problem in drug resistance in environmental isolates in Iran.

  20. Aspergillus, Penicillium and Talaromyces isolated from house dust samples collected around the world

    NARCIS (Netherlands)

    Visagie, C M; Hirooka, Y; Tanney, J B; Whitfield, E; Mwange, K; Meijer, M; Amend, A S; Seifert, K A; Samson, R A

    As part of a worldwide survey of the indoor mycobiota, dust was collected from nine countries. Analyses of dust samples included the culture-dependent dilution-to-extinction method and the culture-independent 454-pyrosequencing. Of the 7 904 isolates, 2 717 isolates were identified as belonging to

  1. Purification and properties of two /beta/-glucosidases isolated from Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Witte, K.; Wartenberg, A.

    1989-01-01

    The cellulase complex of the fungus Aspergillus niger (strain CBS 554.65=ATCC 16 888) was fractionated by gel filtration yielding six pronounced peaks. Only proteins from the fraction corresponding to the first peak (96 kDa) showed /beta/-glucosidase activity vs. the substrate 4-nitrophenyl-/beta/-D-glucopyranoside (pNPG). These proteins have been fractionated by chromatofocusing, yielding two /beta/-glucosidases (I and II) which are shown to be homogeneous in isoelectric focusing experiments (pI=4.6 and 3.8, respectively). Kinetic experiments with pNPG, MU-glucopyranoside and cellobiose revealed that both types of /beta/-glucosidases behave like aryl-/beta/-glucosidases, /beta/-Glucosidase-I acting on pNPG exhibits a split kinetics characterized by high and low substrate-concentration kinetics which are differentiated by different values of V and of K/sub m/. In addition, /beta/-glucosidase-II is shown to be an exo-glucohydrolase as deduced from experiments with MU-cellobiopyranoside. Experimental features should be emphasized; usual soft-gel ion-exchange materials did not work in the chromatofocusing separation of the two /beta/-glucosidases, in contrast to the 10 /mu/-Si 500=DEAE exchange material (Serva) typically used in HPLC-experiments. Furthermore, protein content determinations based on different procedures yielded widely differing values. (orig.).

  2. Antifungal activity of essential oils on Aspergillus parasiticus isolated from peanuts

    Directory of Open Access Journals (Sweden)

    Yooussef Mina M.

    2016-04-01

    Full Text Available Aspergillus parasiticus is one of the most common fungi which contaminates peanuts by destroying peanut shells before they are harvested and the fungus produces aflatoxins. The aim of this study was to evaluate the antifungal activities of seventeen essential oils on the growth of the aflatoxigenic form of A. parasiticus in contaminated peanuts from commercial outlets in Georgia. The agar dilution method was used to test the antifungal activity of essential oils against this form of A. parasiticus at various concentrations: 500; 1,000; 1,500; 2,000; 2,500 ppm. Among the seventeen essential oils tested, the antifungal effect of cinnamon, lemongrass, clove and thyme resulted in complete inhibition of mycelial growth. Cinnamon oil inhibited mycelial growth at ≥ 1,000 ppm, lemongrass and clove oils at ≥ 1,500 ppm and thyme at 2,500 ppm. However, cedar wood, citronella, cumin and peppermint oils showed partial inhibition of mycelial growth. Eucalyptus oil, on the other hand, had less antifungal properties against growth of A. parasiticus, irrespective of its concentration. Our results indicate that the aflatoxigenic form of A. parasiticus is sensitive to selected essential oils, especially cinnamon. These findings clearly indicate that essential oils may find a practical application in controlling the growth of A. parasiticus in stored peanuts.

  3. Rapid detection and typing of pathogenic nonpneumophila Legionella spp. isolates using a multiplex real-time PCR assay.

    Science.gov (United States)

    Benitez, Alvaro J; Winchell, Jonas M

    2016-04-01

    We developed a single tube multiplex real-time PCR assay that allows for the rapid detection and typing of 9 nonpneumophila Legionella spp. isolates that are clinically relevant. The multiplex assay is capable of simultaneously detecting and discriminating L. micdadei, L. bozemanii, L. dumoffii, L. longbeachae, L. feeleii, L. anisa, L. parisiensis, L. tucsonensis serogroup (sg) 1 and 3, and L. sainthelensis sg 1 and 2 isolates. Evaluation of the assay with nucleic acid from each of these species derived from both clinical and environmental isolates and typing strains demonstrated 100% sensitivity and 100% specificity when tested against 43 other Legionella spp. Typing of L. anisa, L. parisiensis, and L. tucsonensis sg 1 and 3 isolates was accomplished by developing a real-time PCR assay followed by high-resolution melt (HRM) analysis targeting the ssrA gene. Further typing of L. bozemanii, L. longbeachae, and L. feeleii isolates to the serogroup level was accomplished by developing a real-time PCR assay followed by HRM analysis targeting the mip gene. When used in conjunction with other currently available diagnostic tests, these assays may aid in rapidly identifying specific etiologies associated with Legionella outbreaks, clusters, sporadic cases, and potential environmental sources. Published by Elsevier Inc.

  4. Changes in qnr Prevalence and Fluoroquinolone Resistance in Clinical Isolates of Klebsiella pneumoniae and Enterobacter spp. Collected from 1990 to 2005▿

    Science.gov (United States)

    Strahilevitz, Jacob; Engelstein, Dalia; Adler, Amos; Temper, Violeta; Moses, Allon E.; Block, Colin; Robicsek, Ari

    2007-01-01

    Clinical isolates of Klebsiella pneumoniae and Enterobacter spp. collected from 1990 through 2005 at a tertiary care center were studied for qnr genes. Isolates bearing these genes emerged in the mid-1990s, coinciding with the time of a rapid increase in fluoroquinolone resistance. Sixty percent of these isolates were ciprofloxacin susceptible by CLSI breakpoints. PMID:17526754

  5. Molecular profiling and bioactive potential of an endophytic fungus Aspergillus sulphureus isolated from Sida acuta: a medicinal plant.

    Science.gov (United States)

    Murali, M; Mahendra, C; Hema, P; Rajashekar, N; Nataraju, A; Sudarshana, M S; Amruthesh, K N

    2017-12-01

    Sida acuta Burm.f. (Malvaceae) extracts are reported to have applications against malaria, diuretic, antipyretic, nervous and urinary diseases. No fungal endophytes of S. acuta are reported. Isolation, identification and evaluation of antibacterial, antioxidant, anticancer and haemolytic potential of fungal endophytes from the ethnomedcinal plant S. acuta. Sida acuta stem segments were placed on PDA medium to isolate endophytic fungi. The fungus was identified by genomic DNA analysis and phylogenetic tree was constructed using ITS sequences (GenBank) to confirm species. The antibacterial efficacy of Aspergillus sulphureus MME12 ethyl acetate extract was tested against Gram-positive and Gram-negative pathogenic bacteria. DPPH free radical scavenging activity, anticancer and DNA fragmentation against EAC cells, and direct haemolytic activity (100-500 μg/mL) using human erythrocytes were determined. The ethyl acetate extract of A. sulphureus (Fresen.) Wehmer (Trichocomaceae) demonstrated significant antibacterial potential against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella typhi compared to streptomycin. MIC against test pathogens was in the range of 15.6-62.5 μg/mL. The antioxidant results revealed significant RSA from 12.43% to 62.02% (IC 50  = 350.4 μg/mL, p ≤ 0.05). MME12 offered considerable inhibition of EAC proliferation (23% to 84%, IC 50  = 216.7 μg/mL, p ≤ 0.05) supported by DNA fragmentation studies. The extract also offered insignificant haemolysis (5.6%) compared to Triton X-100. A single endophytic fungus, A. sulphureus MME12 was isolated and identified using molecular profiling. The above-mentioned findings support the pharmacological application of A. sulphureus MME12 extract and demand for purification of the active principle(s).

  6. Genetic isolation between coastal and fishery-impacted, offshore bottlenose dolphin (Tursiops spp.) populations.

    Science.gov (United States)

    Allen, Simon J; Bryant, Kate A; Kraus, Robert H S; Loneragan, Neil R; Kopps, Anna M; Brown, Alexander M; Gerber, Livia; Krützen, Michael

    2016-06-01

    The identification of species and population boundaries is important in both evolutionary and conservation biology. In recent years, new population genetic and computational methods for estimating population parameters and testing hypotheses in a quantitative manner have emerged. Using a Bayesian framework and a quantitative model-testing approach, we evaluated the species status and genetic connectedness of bottlenose dolphin (Tursiops spp.) populations off remote northwestern Australia, with a focus on pelagic 'offshore' dolphins subject to incidental capture in a trawl fishery. We analysed 71 dolphin samples from three sites beyond the 50 m depth contour (the inshore boundary of the fishery) and up to 170 km offshore, including incidentally caught and free-ranging individuals associating with trawl vessels, and 273 dolphins sampled at 12 coastal sites inshore of the 50 m depth contour and within 10 km of the coast. Results from 19 nuclear microsatellite markers showed significant population structure between dolphins from within the fishery and coastal sites, but also among dolphins from coastal sites, identifying three coastal populations. Moreover, we found no current or historic gene flow into the offshore population in the region of the fishery, indicating a complete lack of recruitment from coastal sites. Mitochondrial DNA corroborated our findings of genetic isolation between dolphins from the offshore population and coastal sites. Most offshore individuals formed a monophyletic clade with common bottlenose dolphins (T. truncatus), while all 273 individuals sampled coastally formed a well-supported clade of Indo-Pacific bottlenose dolphins (T. aduncus). By including a quantitative modelling approach, our study explicitly took evolutionary processes into account for informing the conservation and management of protected species. As such, it may serve as a template for other, similarly inaccessible study populations. © 2016 John Wiley & Sons Ltd.

  7. Prevalence of Listeria spp. and Molecular Characterization of Listeria monocytogenes Isolates from Broilers at the Abattoir.

    Science.gov (United States)

    Bouayad, Leila; Hamdi, Taha M; Naim, Malek; Leclercq, Alexandre; Lecuit, Marc

    2015-07-01

    Products from three broiler abattoirs were sampled for Listeria species to evaluate the changes in the prevalence and contamination rates at two stages of processing. Sampling was performed at the evisceration stage and at the end of processing after packaging and refrigerating at 4°C for 24 h. A total of 212 samples were collected; 52 were from abattoir A, and 80 samples each were collected from abattoirs B and C. Among all samples, 99 (46.7%) tested positive for Listeria, including L. monocytogenes 19 (8.9%), L. innocua 69 (32.5%), L. grayi 10 (4.7%), and L. welshimeri 1 (0.5%). The L. monocytogenes contamination rate varied from 5% to 11.5% in the 3 abattoirs. L. innocua was the most common species identified and was found in 8.8% of the samples from abattoir A and 33.7% of the samples from both abattoirs B and C. Twenty-six of the L. monocytogenes isolates obtained from positive samples were subjected to serotyping by multiplex polymerase chain reaction and characterization by the pulsed-field gel electrophoresis (PFGE) method using two cutting enzymes, ApaI and AscI. Three molecular serogroups were identified: IIa, IIb, and IVb. Serogroup IIa was common to all abattoirs, and serogroups IIb and IVb were found only in abattoir C. The 10 different obtained PFGE profiles were grouped into 7 clusters; some of these clusters were common to the 3 abattoirs, and others were specific to the abattoirs in which they were identified. This study revealed a high prevalence of Listeria spp., particularly L. monocytogenes, in raw broilers. This high incidence presents a risk to consumers due to the potential occurrence of cross-contamination with other foods in domestic refrigerators and the ability of these microorganisms to survive in undercooked products.

  8. Detection of extended spectrum β-lactamase in Pseudomonas spp. isolated from two tertiary care hospitals in Bangladesh

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    Begum Shahanara

    2013-01-01

    Full Text Available Abstract Background Extended spectrum ß-lactamases (ESBLs represent a major group of lactamases responsible for resistance, mostly produced by gram-negative bacteria, to newer generations of ß-lactam drugs currently being identified in large numbers worldwide. The present study was undertaken to see the frequency of ESBL producing Pseudomonas spp. isolated from six hundred clinical specimens (wound, pus, aural, urine, sputum, throat and other swabs collected over a period of three years from two tertiary care hospitals in Bangladesh. Findings Aerobic bacterial culture was performed on aseptically collected swabs and only growth of Pseudomonas was considered for further species identification and ESBL production along with serotyping of Pseudomonas aeruginosa. Antimicrobial susceptibility testing was carried out using the Kirby-Bauer agar diffusion method and ESBL production was detected on Mueller Hinton agar by double-disk synergy technique using Amoxicillin-Clavulanic acid with Ceftazidime, Cefotaxime, Ceftriaxone and Aztreonam. Culture yielded 120 Pseudomonas spp. and 82 of them were biochemically characterized for species. Pseudomonas aeruginosa was found to be the predominant (90.2% species. Of 82 isolates tested for ESBL, 31 (37.8% were ESBL positive with 29 (93.5% as Pseudomonas aeruginosa, the remaining 2 (6.5% were Stenotrophomonas maltophilia and Ralstonia pickettii. Antibiogram revealed Imipenem as the most effective drug (93.3% among all antimicrobials used against Pseudomonas spp. followed by Aminoglycosides (63.7%. Conclusion ESBL producing Pseudomonas spp. was found to be a frequent isolate from two tertiary care hospitals in Bangladesh, showing limited susceptibility to antimicrobials and decreased susceptibility to Imipenem in particular, which is a matter of great concern.

  9. Genetic and Toxigenic Variability within Aspergillus flavus Population Isolated from Maize in Two Diverse Environments in Kenya

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    Sheila Okoth

    2018-01-01

    Full Text Available Aspergillus flavus is the main producer of carcinogenic aflatoxins in agricultural commodities such as maize. This fungus occurs naturally on crops, and produces aflatoxins when environmental conditions are favorable. The aim of this study is to analyse the genetic variability among 109 A. flavus isolates previously recovered from maize sampled from a known aflatoxin-hotspot (Eastern region, Kenya and the major maize-growing area in the Rift Valley (Kenya, and to determine their toxigenic potential. DNA analyses of internal transcribed spacer (ITS regions of ribosomal DNA, partial β-tubulin gene (benA and calmodulin gene (CaM sequences were used. The strains were further analyzed for the presence of four aflatoxin-biosynthesis genes in relation to their capability to produce aflatoxins and other metabolites, targeting the regulatory gene aflR and the structural genes aflP, aflD, and aflQ. In addition, the metabolic profile of the fungal strains was unraveled using state-of-the-art LC-MS/MS instrumentation. The three gene-sequence data grouped the isolates into two major clades, A. minisclerotigenes and A. flavus. A. minisclerotigenes was most prevalent in Eastern Kenya, while A. flavus was common in both regions. A. parasiticus was represented by a single isolate collected from Rift Valley. Diversity existed within the A. flavus population, which formed several subclades. An inconsistency in identification of some isolates using the three markers was observed. The calmodulin gene sequences showed wider variation of polymorphisms. The aflatoxin production pattern was not consistent with the presence of aflatoxigenic genes, suggesting an inability of the primers to always detect the genes or presence of genetic mutations. Significant variation was observed in toxin profiles of the isolates. This is the first time that a profound metabolic profiling of A. flavus isolates was done in Kenya. Positive associations were evident for some metabolites

  10. Genetic and Toxigenic Variability within Aspergillus flavus Population Isolated from Maize in Two Diverse Environments in Kenya.

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    Okoth, Sheila; De Boevre, Marthe; Vidal, Arnau; Diana Di Mavungu, José; Landschoot, Sofie; Kyallo, Martina; Njuguna, Joyce; Harvey, Jagger; De Saeger, Sarah

    2018-01-01

    Aspergillus flavus is the main producer of carcinogenic aflatoxins in agricultural commodities such as maize. This fungus occurs naturally on crops, and produces aflatoxins when environmental conditions are favorable. The aim of this study is to analyse the genetic variability among 109 A. flavus isolates previously recovered from maize sampled from a known aflatoxin-hotspot (Eastern region, Kenya) and the major maize-growing area in the Rift Valley (Kenya), and to determine their toxigenic potential. DNA analyses of internal transcribed spacer (ITS) regions of ribosomal DNA, partial β-tubulin gene (benA) and calmodulin gene (CaM) sequences were used. The strains were further analyzed for the presence of four aflatoxin-biosynthesis genes in relation to their capability to produce aflatoxins and other metabolites, targeting the regulatory gene aflR and the structural genes aflP, aflD, and aflQ. In addition, the metabolic profile of the fungal strains was unraveled using state-of-the-art LC-MS/MS instrumentation. The three gene-sequence data grouped the isolates into two major clades, A. minisclerotigenes and A. flavus . A. minisclerotigenes was most prevalent in Eastern Kenya, while A. flavus was common in both regions. A. parasiticus was represented by a single isolate collected from Rift Valley. Diversity existed within the A. flavus population, which formed several subclades. An inconsistency in identification of some isolates using the three markers was observed. The calmodulin gene sequences showed wider variation of polymorphisms. The aflatoxin production pattern was not consistent with the presence of aflatoxigenic genes, suggesting an inability of the primers to always detect the genes or presence of genetic mutations. Significant variation was observed in toxin profiles of the isolates. This is the first time that a profound metabolic profiling of A. flavus isolates was done in Kenya. Positive associations were evident for some metabolites, while for

  11. [Campylobacter spp.: prevalence and pheno-genotypic characterization of isolates recovered from patients suffering from diarrhea and their pets in La Pampa Province, Argentina].

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    Tamborini, Ana L; Casabona, Luis M; Viñas, María R; Asato, Valeria; Hoffer, Alicia; Farace, María I; Lucero, María C; Corso, Alejandra; Pichel, Mariana

    2012-01-01

    The prevalence of Campylobacter spp. was investigated in 327 patients suffering from diarrhea and in 36 animals (dogs, cats and chickens) owned by the patients that presented infection by Campylobacter in Santa Rosa, La Pampa, Argentina. Campylobacter spp. was isolated in 50/327 patients and in 12/36 animals, being Campylobacter jejuni the most common species. Resistance to ciprofloxacin (65 %) and tetracycline (32 %) was found among 35 isolates of human origin studied. Seven genetic subtypes were observed among 13 C. jejuni isolates by pulsed field gel electrophoresis. Two subtypes grouped isolates belonging to patients and their respective dogs whereas another subtype grouped one isolate of human origin and two isolates from the patient's chickens. The results of this investigation highlight the need to strengthen surveillance of Campylobacter spp. not only in poultry, which is recognized as the main reservoir, but also in pets, which were shown to be asymptomatic carriers of the pathogen.

  12. Prevalence, identification by a DNA microarray-based assay of human and food isolates Listeria spp. from Tunisia.

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    Hmaïed, F; Helel, S; Le Berre, V; François, J-M; Leclercq, A; Lecuit, M; Smaoui, H; Kechrid, A; Boudabous, A; Barkallah, I

    2014-02-01

    We aimed at evaluating the prevalence of Listeria species isolated from food samples and characterizing food and human cases isolates. Between 2005 and 2007, one hundred food samples collected in the markets of Tunis were analysed in our study. Five strains of Listeria monocytogenes responsible for human listeriosis isolated in hospital of Tunis were included. Multiplex PCR serogrouping and pulsed field gel electrophoresis (PFGE) applying the enzyme AscI and ApaI were used for the characterization of isolates of L. monocytogenes. We have developed a rapid microarray-based assay to a reliable discrimination of species within the Listeria genus. The prevalence of Listeria spp. in food samples was estimated at 14% by using classical biochemical identification. Two samples were assigned to L. monocytogenes and 12 to L. innocua. DNA microarray allowed unambiguous identification of Listeria species. Our results obtained by microarray-based assay were in accordance with the biochemical identification. The two food L. monocytogenes isolates were assigned to the PCR serogroup IIa (serovar 1/2a). Whereas human L. monocytogenes isolates were of PCR serogroup IVb, (serovars 4b). These isolates present a high similarity in PFGE. Food L. monocytogenes isolates were classified into two different pulsotypes. These pulsotypes were different from that of the five strains responsible for the human cases. We confirmed the presence of Listeria spp. in variety of food samples in Tunis. Increased food and clinical surveillance must be taken into consideration in Tunisia to identify putative infections sources. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  13. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

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    Paulo S. Monteiro

    2015-03-01

    Full Text Available An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

  14. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

    Science.gov (United States)

    Monteiro, Paulo S.; Guimarães, Valéria M.; de Melo, Ricardo R.; de Rezende, Sebastião T.

    2015-01-01

    An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for sodium phytate hydrolysis was 30.9 mM, while the k cat and k cat / K M were 1.46 ×10 5 s −1 and 4.7 × 10 6 s −1 .M −1 , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg 2+ , Cd 2+ , K + and Ca 2+ , and it was drastically inhibited by F − . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment. PMID:26221114

  15. Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

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    Colla, Luciane Maria; Ficanha, Aline M. M.; Rizzardi, Juliana; Bertolin, Telma Elita; Reinehr, Christian Oliveira; Costa, Jorge Alberto Vieira

    2015-01-01

    Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through submerged fermentation presented optimal activities at 37°C and pH 7.2 and those obtained through solid-state fermentation at 35°C and pH 6.0. The enzymes produced by submerged fermentation were more temperature-stable than those obtained by solid-state fermentation, presenting 72% of residual activity after one hour of exposition at 90°C. Lipases obtained through submerged fermentation had 80% of stability in acidic pH and those obtained through solid-state fermentation had stability greater than 60% in alkaline pH. PMID:26180809

  16. Phytosterols elevation in bamboo shoot residue through laboratorial scale solid-state fermentation using isolated Aspergillus niger CTBU.

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    Zheng, X X; Chen, R S; Shen, Y; Yin, Z Y

    2014-04-01

    Aspergillus niger CTBU isolated from local decayed bamboo shoot residue was employed to solid-state fermentation (SSF) of bamboo shoot residue to elevate the content of phytosterols. Strain acclimatization was carried out under the fermentation condition using bamboo shoot as substrate for fermentation performance improvement. The optimal fermentation temperature and nitrogen level were investigated using acclimatized strain, and SSF was carried out in a 500-ml Erlenmeyer flask feeding 300-mg bamboo shoot residue chips under the optimal condition (33 °C and feeding 4 % urea), and 1,186 mg (100 g)(-1) of total phytosterol was attained after 5-day fermentation, in comparison, only 523 mg (100 g)(-1) of phytosterol was assayed in fresh shoots residue. HPLC analysis of the main composition of total phytosterols displays that the types of phytosterols and composition ratio of main sterols keep steady. This laboratorial scale SSF unit could be scaled up for raw phytosterols production from discarded bamboo shoot residue and could reduce its cost.

  17. Characterization of a Newly Isolated Marine Fungus Aspergillus dimorphicus for Optimized Production of the Anti-Tumor Agent Wentilactones

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    Rui Xu

    2015-11-01

    Full Text Available The potential anti-tumor agent wentilactones were produced by a newly isolated marine fungus Aspergillus dimorphicus. This fungus was derived from deep-sea sediment and identified by polyphasic approach, combining phenotypic, molecular, and extrolite profiles. However, wentilactone production was detected only under static cultures with very low yields. In order to improve wentilactone production, culture conditions were optimized using the response surface methodology. Under the optimal static fermentation conditions, the experimental values were closely consistent with the prediction model. The yields of wentilactone A and B were increased about 11-fold to 13.4 and 6.5 mg/L, respectively. The result was further verified by fermentation scale-up for wentilactone production. Moreover, some small-molecule elicitors were found to have capacity of stimulating wentilactone production. To our knowledge, this is first report of optimized production of tetranorlabdane diterpenoids by a deep-sea derived marine fungus. The present study might be valuable for efficient production of wentilactones and fundamental investigation of the anti-tumor mechanism of norditerpenoids.

  18. Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

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    Luciane Maria Colla

    2015-01-01

    Full Text Available Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through submerged fermentation presented optimal activities at 37°C and pH 7.2 and those obtained through solid-state fermentation at 35°C and pH 6.0. The enzymes produced by submerged fermentation were more temperature-stable than those obtained by solid-state fermentation, presenting 72% of residual activity after one hour of exposition at 90°C. Lipases obtained through submerged fermentation had 80% of stability in acidic pH and those obtained through solid-state fermentation had stability greater than 60% in alkaline pH.

  19. In vitro probiotic potential of Lactobacillus spp. isolated from fermented milks

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    A.F. Cunha

    2013-12-01

    Full Text Available The potential of in vitro probiotic Lactobacillus spp. was evaluated in fermented milks marketed in Belo Horizonte, MG, Brazil. Of the samples analyzed, 86.7% had at least 10(6 CFU/mL of Lactobacillus spp., complying with the Brazilian quality standards for fermented milks. Furthermore, 56.7% had minimum count ranging from 10(8 to 10(9 CFU/mL, which is in accordance with legal parameters. The remaining 43.3% would not be able to satisfactorily guarantee benefits to consumers. The amount of Lactobacillus spp. varied between batches of products, which may indicate failures in monitoring during manufacture, transport or storage. All strains of Lactobacillus spp. showed some inhibitory activity against the indicator microorganisms, being more pronounced against pathogenic microorganisms than against non-pathogenic (P<0.05. Samples of Lactobacillus spp. showed different profiles of antimicrobial susceptibility, with an occurrence of cases of multidrug resistance. All strains tested showed sensitivity to bile salts (0.3% and resistance to gastric pH (2.0. Lactobacillus spp. of commercial fermented milks should be present in higher amounts in some brands, be resistant to bile salts and have no multiple resistance to antimicrobials.

  20. Resistance to antimicrobials and biofilm formation in Staphylococcus spp. isolated from bovine mastitis in the Northeast of Brazil.

    Science.gov (United States)

    da Costa Krewer, Carina; Santos Amanso, Evandro; Veneroni Gouveia, Gisele; de Lima Souza, Renata; da Costa, Mateus Matiuzzi; Aparecido Mota, Rinaldo

    2015-03-01

    Mastitis is the principal disease affecting dairy herds worldwide. The aim of the present study was to characterize phenotypic and genotypic features associated with resistance to antimicrobials in Staphylococcus spp. isolated from 2064 milk samples of 525 lactating cows in the Northeast of Brazil. Of the 218 isolates analyzed, 57.8% were characterized as Staphylococcus aureus, 28% as coagulase-positive staphylococci other than S. aureus (oCPS), and 14.2% as coagulase-negative staphylococci (CNS). The test for susceptibility to antimicrobials showed amoxicillin (32.6%) to be the less effective drug in vitro, and the multi-drug resistance (MDR) rate for beta-lactams varied from 0 to 0.75. The genotypic characterization showed that 93.1% of the samples were tested positive for the blaZ gene, while none amplified mecA. The antibiotic efflux mechanism was observed in 0.9% of isolates. The biofilm formation was found in 3.7 and 96.3% of samples, respectively, on Congo red agar and on the microplate adhesion test, while the icaD gene was present in 92.2% of Staphylococcus spp. The high frequency of blaZ gene observed in this study was associated with the resistance of most Staphylococcus spp. to one or more of the beta-lactams tested, which are routinely used in Brazilian herds for mastitis treatment. The biofilm formation was also detected in the isolates analyzed being an important characteristic for pathogenicity and antimicrobial resistance of bacteria.

  1. Study of Inhibitory Effect of Echinops cephalotes on Candida Spp. Isolated from Vulvovaginal Candidiasis Patients in Isfahan

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    Sara Heshmati

    2016-06-01

    Full Text Available Background Candida spp. has been considered as the agents of acute and recurrent vulvovaginal candidiasis. Objectives The aim of current study was the evaluation of antifungal activity of Echinops cephalotes (Leaves and stem, manna plant against species of Candida isolated from patients with vulvovaginal candidiasis. Materials and Methods In this research study identification of clinical isolates (50 cases was inducted to the species level by means of conventional mycological methods, morophology on corn meal agar and chromogenic agar, germ tube production and biochemical methods. Antifungal activity of the ethanolic, methanolic and aqueous extracts of E. cephalotes was studied against isolated Candida using agar well diffusion and microdilution methods. Results Candida spp. which isolated from patients was C. albicans, C. glabrata, C. tropicalis and C. parapsilosis. The inhibition zone of ethanolic extract was 16.6, 13.3, 14, and 22 mg/mL respectively. Minimum inhibitory concentration (MIC for most the cases were 15.6 mg/mL. The inhibition zone of aqueous extract was 16.8, 16.7, 15 and 15 mg/mL respectively. MIC for most the cases were 15.6-31.2 mg/mL. The inhibition zone of methanolic extract was 15.4, 13.2, 12 and 18 respectively. MIC for most of the cases was 7.8 mg/mL. Among the different extracts, ethanolic extract has the highest and aqueous extract has the lowest anti-Candida activity. Ethanolic, methanolic and aqueous extracts of trehala manna did not show any antifungal activity. Conclusions This research is the first study on antifungal activity of E. cephalotes. Hence, this plant may be used further as medicinal plant against Candida spp.

  2. Dissimilatory nitrate reduction by Aspergillus terreus isolated from the seasonal oxygen minimum zone in the Arabian Sea.

    Science.gov (United States)

    Stief, Peter; Fuchs-Ocklenburg, Silvia; Kamp, Anja; Manohar, Cathrine-Sumathi; Houbraken, Jos; Boekhout, Teun; de Beer, Dirk; Stoeck, Thorsten

    2014-02-11

    A wealth of microbial eukaryotes is adapted to life in oxygen-deficient marine environments. Evidence is accumulating that some of these eukaryotes survive anoxia by employing dissimilatory nitrate reduction, a strategy that otherwise is widespread in prokaryotes. Here, we report on the anaerobic nitrate metabolism of the fungus Aspergillus terreus (isolate An-4) that was obtained from sediment in the seasonal oxygen minimum zone in the Arabian Sea, a globally important site of oceanic nitrogen loss and nitrous oxide emission. Axenic incubations of An-4 in the presence and absence of oxygen and nitrate revealed that this fungal isolate is capable of dissimilatory nitrate reduction to ammonium under anoxic conditions. A ¹⁵N-labeling experiment proved that An-4 produced and excreted ammonium through nitrate reduction at a rate of up to 175 nmol ¹⁵NH₄⁺ g⁻¹ protein h⁻¹. The products of dissimilatory nitrate reduction were ammonium (83%), nitrous oxide (15.5%), and nitrite (1.5%), while dinitrogen production was not observed. The process led to substantial cellular ATP production and biomass growth and also occurred when ammonium was added to suppress nitrate assimilation, stressing the dissimilatory nature of nitrate reduction. Interestingly, An-4 used intracellular nitrate stores (up to 6-8 μmol NO₃⁻ g⁻¹ protein) for dissimilatory nitrate reduction. Our findings expand the short list of microbial eukaryotes that store nitrate intracellularly and carry out dissimilatory nitrate reduction when oxygen is absent. In the currently spreading oxygen-deficient zones in the ocean, an as yet unexplored diversity of fungi may recycle nitrate to ammonium and nitrite, the substrates of the major nitrogen loss process anaerobic ammonium oxidation, and the potent greenhouse gas nitrous oxide.

  3. In vitro and in vivo effects of Pseudomonas spp. and Bacillus sp. on Fusarium acuminatum, Botrytis cinerea and Aspergillus niger infecting cucumber

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    Jasmina Zdravković

    2015-09-01

    Full Text Available Cucumber (Cucumis sativus L is an important member of the Cucurbitaceae family. Production of healthy nursery is necessary for high-quality production of this crop in greenhouses and in fields. With the idea of minimizing the use of pesticides and mineral fertilizers to preserve soil quality, we investigated the effects of plant growth promoting bacteria (PGPB on growth promotion and protection of cucumber plants from phytopathogenic fungi. The effects of Pseudomonas spp. strains with different antifungal activities and Bacillus sp. Q10 strain with PGP activity were tested on cucumber plants. Antagonistic activity of Pseudomonas spp. against the growth of several phytopathogenic fungi isolated from cucumber: F. acuminatum, B. cinerea and A. niger, was observed. The influences of overnight cultures, supernatants and heat-stable antifungal factors were tested on the phytopathogenic fungi in vitro. Pseudomonas sp. K35 and K24 strains were more effective than P. chlororaphis Q16 and Pseudomonas sp. K27, showing 70-80% of fungal growth inhibition regardless of culture or fraction applied. The good antagonists that belong to pseudomonads and Bacillus sp. Q10 strain were used as mixtures for estimation of plant growth and health promoting effects on cucumber plants. Growth dynamics differed depending on the applied strain of Pseudomonas sp. The M3 treatment (a mixture of Bacillus sp. Q10 and P. chlororaphis Q16 stimulated the initial phase of growth, while M4 (a mixture of Bacillus sp. Q10 and Pseudomonas sp. K24 resulted in the maximal height at the final measurement. Significant differences in leaf and plant weight (M4, and leaf weight (M5, containing K35 strain were found after the treatments. No significant differences in chlorophyll and NBI level were observed in any of the tested combinations. The obtained results suggested that M3 was suitable for stimulation of the early phase of cucumber growth, while the mixtures M4 and M5 improved plant

  4. Isolation, screening and characterization of a novel extracellular xylanase from Aspergillus niger (KP874102.1) and its application in orange peel hydrolysis.

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    Uday, Uma Shankar Prasad; Majumdar, Ria; Tiwari, Onkar Nath; Mishra, Umesh; Mondal, Abhijit; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2017-12-01

    In the present work, a potent xylanase producing fungal strain Aspergillus niger (KP874102.1) was isolated through cultural and morphological observations from soil sample of Baramura forest, Tripura west, India. 28S rDNA technique was applied for genomic identification of this fungal strain. The isolated strain was found to be phylogenetically closely related to Aspergillus niger. Kinetic constants such as K m and V max for extracellular xylanase were determined using various substrate such as beech wood xylan, oat spelt xylan and CM cellulose through Lineweaver-Burk plot. K m , V max and K cat for beech wood xylan are found to be 2.89mg/ml, 2442U and 426178Umlmg -1 respectively. Crude enzyme did not show also CM cellulose activity. The relative efficiency of oat spelt xylan was found to be 0.819 with respect to beech wood xylan. After acid hydrolysis, enzyme was able to produce reducing sugar with 17.7, 35.5, 50.8 and 65% (w/w) from orange peel after 15, 30, 45 and 60min incubation with cellulase free xylanase and maximum reducing sugar formation rate was found to be 55.96μg/ml/min. Therefore, the Aspergillus niger (KP874102.1) is considered as a potential candidate for enzymatic hydrolysis of orange peel. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Reduced susceptibility of Enterococcus spp. isolates from Cairo University Hospital to tigecycline: Highlight on the influence of proton pump inhibitors.

    Science.gov (United States)

    Hassan, Reem Mostafa; Ghaith, Doaa Mohammad; Ismail, Dalia Kadry; Zafer, Mai Mahmoud

    2018-03-01

    The incidence of reduced susceptibility to tigecycline (TIG) is increasing. This study aimed to analyse the in vitro activity of TIG against Enterococcus spp. isolates recovered from hospitalised patients and to evaluate the effect of omeprazole on the in vitro antimicrobial activity of TIG against several enterococcal species. A total of 67 Enterococcus clinical isolates were identified by MALDI-TOF/MS and multiplex PCR. Minimum inhibitory concentrations (MICs) of TIG alone and in combination with omeprazole (10, 30 and 60mg/L) were determined by broth microdilution. Antibiotic susceptibility to other antibiotics was determined by disk diffusion. The presence of van, tet(X) and tet(X1) genes was tested by multiplex PCR. Of the 67 Enterococcus isolates, 2 (3.0%) were resistant to TIG and 13 (19.4%) were intermediate-resistant according to EUCAST. The frequencies of resistance to norfloxacin (80.6%), doxycycline (80.6%), levofloxacin (74.6%) and ciprofloxacin (71.6%) were highest, whilst that of vancomycin (25.4%) was lowest. The vanA gene was detected in 11 Enterococcus isolates (8 Enterococcus faecalis, 3 Enterococcus faecium), vanB in 3 Enterococcus isolates (2 E. faecium, 1 E. faecalis) and vanC-2/3 in 3 Enterococcus casseliflavus. Nine isolates (13.4%) were positive for tet(X1). TIG resistance occurred both in patients receiving or not TIG and/or omeprazole. Omeprazole increased TIG MICs by 4-128-fold. The possibility of selection of TIG-non-susceptible Enterococcus in the gut may occur with long-term use of omeprazole. Omeprazole influenced TIG activity in a concentration-dependent manner. To our knowledge; this is the first report of TIG-non-susceptible Enterococcus spp. in Egypt. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  6. Enterotoxigenicidade de Staphylococcus spp. isolados de leite in natura Enterotoxigenicity of Staphylococcus spp. isolated of milk in natura

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    Tânia Lúcia Montenegro Stamford

    2006-03-01

    agents, which can take to manifested diseases by action of pathogenic microorganisms or their toxins. It was researched the occurrence of strains of Staphylococcus and your capacity in producing enterotoxins in milk in natura, that have been produced or commercialized in the State Pernambuco, Brazil. 109 strains of Staphylococus positive and negative coagulase of milk in natura were selected. The identification of the isolated strains was accomplished through morphologic and biochemical tests as: catalase, coagulase, haemolysins, DNAse, thermonuclease, acetoin production (VP and carbohydrates metabolism (glucose, maltose and mannitol. From the 77 coagulase positive strains 30 were identified as S. aureus, 3 as S. hyicus, 16 as S. intermedius, 13 as S. aureus identification presumptive and 15 as SCP. Among 32 coagulase negative strains 2 were identified as S. capitis, 1 as S. carnosus, 6 as S. chromogenes, 1 as S. hyicus, 1 as S. schleiferi and 21 as SCN. Fourty-three strains that presented very evident thermonuclease reaction, were selected in order to perform for staphylococcal enterotoxins analysis by the immuno enzimatic test (ELFA. 10 strains showed negative reaction for enterotoxins: S. aureus (4, S. carnosus (1, S. chromogenes (2, S. hyicus (2 and S. intermedius (1. Strains that gave positive results, were S. aureus (17, S. chromogenes (2, S. hyicus (1, S. intermedius (8, S. aureus identified presumptively (2 and of the groups SCP (1 and SCN (2. The species that presented larger number of enterotoxigenics strains were S. aureus and S. intermedius. Results can be attributed to the inadequate manipulation or food recontamination during the storage and distribution.

  7. Use of cell surface protein typing for genotyping of azole-resistant and -susceptible Aspergillus fumigatus isolates in Iran

    NARCIS (Netherlands)

    Falahatinejad, M.; Vaezi, A.; Fakhim, H.; Abastabar, M.; Shokohi, T.; Zahedi, N.; Ansari, S.; Meis, J.F.G.M.; Badali, H.

    2018-01-01

    Aspergillus fumigatus is the leading cause of mortality in severely immunocompromised individuals. Understanding pathogen dispersion and relatedness is essential for determining the epidemiology of nosocomial infections. Therefore, the aim of this study was to investigate the diversity and putative

  8. Prevalence and Antibiotic Resistance of Listeria Spp. Isolated from Ready-to-Eat Foods in Ankara

    OpenAIRE

    ŞİRELİ, Ufuk Tansel; GÜCÜKOĞLU, Ali

    2014-01-01

    In this study the presence of Listeria spp. is tested in 100 ready-to-eat food samples purchased from different stores and traditional food shops in the province of Ankara. The tested materials were 20 each of the following: mayonnaise based salad, kadınbudu köfte (fried meatball), fried liver, rice stuffed mussel, and green salad. Microbiological analyzes showed that 13 of 100 salad samples (13%) were contaminated with Listeria spp. while 10 of 100 salad samples (10%) were contaminated with ...

  9. Chemotactic responses to gas oil of Halomonas spp. strains isolated from saline environments in Argentina Respuesta quimiotáctica hacia gas oil de cepas de Halomonas spp. aisladas de ambientes salinos de Argentina

    Directory of Open Access Journals (Sweden)

    Sebastián D´Ippólito

    2011-06-01

    Full Text Available In this study, two halophilic bacterial strains isolated from saline habitats in Argentina grew in the presence of gas oil. They were identified as Halomonas spp. and Nesterenkonia sp. by 16S ribosomal RNA sequencing. Chemotaxis towards gas oil was observed in Halomonas spp. by using swimming assays.En el presente trabajo se aislaron dos cepas bacterianas halofílicas a partir de muestras obtenidas en ambientes salinos de Argentina, que crecieron en presencia de gasoil como única fuente de carbono. Las cepas aisladas se identificaron como Halomonas spp. y Nesterenkonia sp. mediante secuenciación del gen del ARN ribosomal 16S. En ensayos de swimming, las cepas del genero Halomonas spp. mostraron una respuesta quimiotáctica hacia el gas oil.

  10. A New Biocontrol Fungus Trichoderma Kongii in the Kingdom of Saudi Arabia Isolation and Identification

    International Nuclear Information System (INIS)

    AlYahya, F.A; ElHussieni, S.M; Ibrahim, A.A; Ibrahim, Y.E

    2007-01-01

    A total of 164 soil and root samples of different plant groups were collected from Abu-Arish governorate , Jazan province South West Saudi Arabia during the period of 2004-2005. Each sample contained feeder roots and approximately 250 g soil, taken from a depth of 20 cm of the soil surface. Samples were analyzed by two different media. Culture fungi on Malt Extract Agar identified by Biolog Systems and culture fungi on Potato Dextrose Agar medium containing chloramphenicol were identified by microscopic characterization. Results showed that, 11 different types of fungi isolated from tested samples, Fusarium spp (40%), Rhizoctonia solani,(12%) Trichoderma spp (12%), Macrophomina phaseoina. (7 %), Aspergillus spp (18 %) were the predominant fungal species. Helminthosporium spp (3%), Alternaria alternate (2%), Pythium spp (2%), Curviularia spp (2%), Cladsporium spp. (1%) and Mucor spp. (1%) were less frequent. (author)

  11. [Determination of in vitro susceptibilities of Brucella spp. strains against 11 different antibacterial gents isolated from blood cultures].

    Science.gov (United States)

    Keşli, Recep; Bilgin, Hüseyin; Yılmaz, Halim

    2017-07-01

    Brucellosis is a worldwide zoonotic disease and still continuous to be a major public health problem. In this study, it was aimed to identify the Brucella strains to the species level isolated from blood cultures, and to determine the rate of antimicrobial susceptibility against eleven antibacterial agents. A total of 106 Brucella spp. strains were included in the study, which were isolated from blood cultures in University of Health Sciences, Konya Training and Research Hospital, Medical Microbiology Laboratory between January 2011 and June 2013. Identification of the isolated strains were mainly based on conventional methods. In vitro antibacterial susceptibilities of azithromycin, ciprofloxacin, doxycycline, gentamicin, levofloxacin, moxifloxacin, rifampicin, streptomycin, tetracycline, tigecycline, and trimethoprim/sulfamethoxazole, were evaluated by using the gradient (E-test, bioMerieux, France) strip method. The bacterial suspensions adjusted to 0.5 McFarland turbidity was inoculated to Mueller Hinton agar plates, supplemented with 5% sheep blood, and E-test strips of selected antibacterial were applied. The plates were incubated in ambient air 48 hours at 37ºC and Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 were used as quality control strains for antimicrobial susceptibility testing. Minimum inhibitors concentration (MIC) values were interpreted according to Clinical and Laboratory Standards Institute (CLSI) guidelines for slow-growing bacteria such as Haemophilus spp. Of the 106 Brucella spp. strains included in to the study, 90 were identified as Brucella melitensis, and 16 were Brucella abortus. MIC90 values of azithromycin, ciprofloxacin, doxycycline, gentamicin, levofloxacin, moxifloxacin, rifampicin, streptomycin, tetracycline, tigecycline, and trimethoprim/sulfamethoxazole were determined as 1 µg/ml, 0.25 µg/ml, 0.19 µg/ml, 0.25 µg/ml, 0.19 µg/ml, 0.75 µg/ml, 0.25 µg/ml, 0.75 µg/ml, 0.38 µg/ml, 0.64 µg/ml, and 0

  12. LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) FOR THE DETECTION OF SALMONELLA SPP. ISOLATED FROM DIFFERENT FOOD TYPES

    OpenAIRE

    Kostas Papanotas; Petros A. Kokkinos; Panos G. Ziros; Apostolos Vantarakis

    2012-01-01

    The objective of this study was the application and evaluation of a loop-mediated isothermal amplification (LAMP) method for the detection of Salmonella spp. strains isolated from food samples. Salmonella specific invA gene sequences (50 strains, 15 serotypes) were amplified at 65oC in 60 min. All of the strains of Salmonella subsp. Enterica were shown to be positive using the LAMP reaction assay, whereas, all other bacteria, virus and yeasts tested in this study were negative. LAMP products ...

  13. optimization of process parameters for lovastatin production under solid-state fermentation from ground corn cobs by gamma irradiated aspergillus tamarri isolate

    International Nuclear Information System (INIS)

    Mattar, Z.A.; Khalaf, M.A.; Meleigy, S.A.

    2010-01-01

    rapid screening method is demonstrated for isolating lovastatin overproducing strains of gamma irradiated aspergillus tamarri. the screening methodology, based on the activity of lovastatin against the yeast candida albicans. among 24 gamma irradiated isolates of a. tamarri, the isolate G-8 was selected as best producer for lovastatin. solid state fermentation (SSF)was evaluated to produce lovastatin by a. tamarri G-8 isolate using ground corn cobs as substrate. monofactorial experiments were adopted to optimize the fermentation conditions. various crucial parameters such as particle size, moisture content, ph, temperature, inoculum size and incubation time were derived. corn cobs of particle size 0.4 mm having moisture level of 60 % and ph 5 gave the highest yield of lovastatin (12.4 mg/gram dry substrate) when inoculated with a. tamarri G-8 at inoculum size 10 % and 28 degree C for 8 days.

  14. The tetracycline resistance determinant Tet 39 and the sulphonamide resistance gene sulII are common among resistant Acinetobacter spp. isolated from integrated fish farms in Thailand

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Petersen, Andreas

    2007-01-01

    Objectives: To determine the genetic basis for tetracycline and sulphonamide resistance and the prevalence of class I and II integrons in oxytetracycline-resistant Acinetobacter spp. from integrated fish farms in Thailand. Methods: A total of 222 isolates were screened for tetracycline resistance...... and Southern blots with sulII and tet(39) probes were performed on selected isolates. Results: The recently identified tetracycline resistance gene tet(39) was demonstrated in 75% (166/222) of oxytetracycline-resistant Acinetobacter spp. from integrated fish farms in Thailand. Isolates that were also...

  15. Primary Isolation and Characterization of Tenacibaculum maritimum from Chilean Atlantic Salmon Mortalities Associated with a Pseudochattonella spp. Algal Bloom.

    Science.gov (United States)

    Apablaza, Patricia; Frisch, Kathleen; Brevik, Øyvind Jakobsen; Småge, Sverre Bang; Vallestad, Camilla; Duesund, Henrik; Mendoza, Julio; Nylund, Are

    2017-09-01

    This study presents the first isolation of Tenacibaculum maritimum from farmed Atlantic Salmon Salmo salar in Chile. The isolate, designated T. maritimum Ch-2402, was isolated from gills of Atlantic Salmon at a farm located in region X, Los Lagos, Chile, during the harmful algal bloom caused by Pseudochattonella spp. in February 2016. The algal bloom is reported to have caused 40,000 metric tons of mortality in this salmon farming area. The bacterium T. maritimum, which causes tenacibaculosis, is recognized as an important pathogen of marine fish worldwide. Genetic, phylogenetic, and phenotypic characterizations were used to describe the T. maritimum Ch-2402 isolate. The isolate was similar to the type strain of T. maritimum but was genetically unique. Tenacibaculum dicentrarchi isolates were also recovered during sampling from the same farm. Based on the fact that T. maritimum has been shown to cause disease in Atlantic Salmon in other regions, the presence of this bacterium poses a potential risk of disease to fish in the Chilean aquaculture industry. Received November 6, 2016; accepted May 29, 2017.

  16. Polyphasic approach to the identification and characterization of aflatoxigenic strains of Aspergillus section Flavi isolated from peanuts and peanut-based products marketed in Malaysia.

    Science.gov (United States)

    Norlia, M; Jinap, S; Nor-Khaizura, M A R; Son, R; Chin, C K; Sardjono

    2018-05-31

    Peanuts are widely consumed as the main ingredient in many local dishes in Malaysia. However, the tropical climate in Malaysia (high temperature and humidity) favours the growth of fungi from Aspergillus section Flavi, especially during storage. Most of the species from this section, such as A. flavus, A. parasiticus and A. nomius, are natural producers of aflatoxins. Precise identification of local isolates and information regarding their ability to produce aflatoxins are very important to evaluate the safety of food marketed in Malaysia. Therefore, this study aimed to identify and characterize the aflatoxigenic and non-aflatoxigenic strains of Aspergillus section Flavi in peanuts and peanut-based products. A polyphasic approach, consisting of morphological and chemical characterizations was applied to 128 isolates originating from raw peanuts and peanut-based products. On the basis of morphological characters, 127 positively identified as Aspergillus flavus, and the other as A. nomius. Chemical characterization revealed six chemotype profiles which indicates diversity of toxigenic potential. About 58.6%, 68.5%, and 100% of the isolates are positive for aflatoxins, cyclopiazonic acid and aspergillic acid productions respectively. The majority of the isolates originating from raw peanut samples (64.8%) were aflatoxigenic, while those from peanut-based products were less toxigenic (39.1%). The precise identification of these species may help in developing control strategies for aflatoxigenic fungi and aflatoxin contamination in peanuts, especially during storage. These findings also highlight the possibility of the co-occurrence of other toxins, which could increase the potential toxic effects of peanuts. Copyright © 2018. Published by Elsevier B.V.

  17. Antimicrobial susceptibility and antibiotic resistance gene transfer analysis of foodborne, clinical, and environmental Listeria spp. isolates including Listeria monocytogenes.

    Science.gov (United States)

    Bertsch, David; Muelli, Mirjam; Weller, Monika; Uruty, Anaïs; Lacroix, Christophe; Meile, Leo

    2014-02-01

    The aims of this study were to assess antibiotic resistance pheno- and genotypes in foodborne, clinical, and environmental Listeria isolates, as well as to elucidate the horizontal gene transfer potential of detected resistance genes. A small fraction of in total 524 Listeria spp. isolates (3.1%) displayed acquired antibiotic resistance mainly to tetracycline (n = 11), but also to clindamycin (n = 4) and trimethoprim (n = 3), which was genotypically confirmed. In two cases, a tetracycline resistance phenotype was observed together with a trimethoprim resistance phenotype, namely in a clinical L. monocytogenes strain and in a foodborne L. innocua isolate. Depending on the applied guidelines, a differing number of isolates (n = 2 or n = 20) showed values for ampicillin that are on the edge between intermediate susceptibility and resistance. Transferability of the antibiotic resistance genes from the Listeria donors, elucidated in vitro by filter matings, was demonstrated for genes located on transposons of the Tn916 family and for an unknown clindamycin resistance determinant. Transfer rates of up to 10(-5) transconjugants per donor were obtained with a L. monocytogenes recipient and up to 10(-7) with an Enterococcus faecalis recipient, respectively. Although the prevalence of acquired antibiotic resistance in Listeria isolates from this study was rather low, the transferability of these resistances enables further spread in the future. This endorses the importance of surveillance of L. monocytogenes and other Listeria spp. in terms of antibiotic susceptibility. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  18. Secreted aspartate proteinases, a virulence factor of Candida spp.: Occurrence among clinical isolates

    Czech Academy of Sciences Publication Activity Database

    Hamal, P.; Dostál, Jiří; Raclavský, V.; Krylová, M.; Pichová, Iva; Hrušková-Heidingsfeldová, Olga

    2004-01-01

    Roč. 49, č. 4 (2004), s. 491-496 ISSN 0015-5632 R&D Projects: GA MZd NI6485 Institutional research plan: CEZ:AV0Z4055905 Keywords : Candida spp. * aspartate proteinases * RAPD typing Subject RIV: CE - Biochemistry Impact factor: 1.034, year: 2004

  19. Isolation of Campylobacter spp and Escherichia coli 0157: H7 from ...

    African Journals Online (AJOL)

    Objectives: To determine the biosafety of a free range indigenous chicken value chain with reference to zoonotic bacteria, Campylobacter spp and Escherichia coli 0157: H7. Design: cross-sectional sampling of chickens and chicken meat carcasses at farm and market level. Setting: Makueni and Nairobi Counties. Subjects: ...

  20. Isolation, Purification, and Identification of Taxol and Related Taxanes from Taxol-Producing Fungus Aspergillus niger subsp. taxi.

    Science.gov (United States)

    Li, Dan; Fu, Dongwei; Zhang, Yue; Ma, Xueling; Gao, Liguo; Wang, Xioahua; Zhou, Dongpo; Zhao, Kai

    2017-08-28

    The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by 1 H NMR, 13 C NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (β-sitosterol and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger , which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger .

  1. Alkaline thermostable pectinase enzyme from Aspergillus niger strain MCAS2 isolated from Manaslu Conservation Area, Gorkha, Nepal.

    Science.gov (United States)

    Khatri, Bhim Prakash; Bhattarai, Tribikram; Shrestha, Sangita; Maharjan, Jyoti

    2015-01-01

    Pectinase enzymes are one of the commercially important enzymes having great potential in various industries especially in food industry. Pectinases accounts for 25 % of global food enzymes produced and their market is increasing day by day. Therefore, the exploration of microorganism with novel characteristics has always been the focus of the research. Microorganism dwelling in unique habitat may possess unique characteristics. As such, a pectinase producing fungus Aspergillus niger strain MCAS2 was isolated from soil of Manaslu Conservation Area (MCA), Gorkha, Nepal. The optimum production of pectinase enzyme was observed at 48 h of fermentation. The pectinase enzyme was partially purified by cold acetone treatment followed by Sephadex G-75 gel filtration chromatography. The partially purified enzyme exhibited maximum activity 60 U/mg which was almost 8.5-fold higher than the crude pectinase. The approximate molecular weight of the enzyme was found to be 66 kDa as observed from SDS-PAGE. The pectinase enzyme was active at broad range of temperature (30-70 °C) and pH (6.2-9.2). Optimum temperature and pH of the pectinase enzyme were 50 °C and 8.2 respectively. The enzyme was stable up to 70 °C and about 82 % of pectinase activity was still observed at 100 °C. The thermostable and alkaline nature of this pectinase can meet the demand of various industrial processes like paper and pulp industry, in textile industry, fruit juice industry, plant tissue maceration and wastewater treatment. In addition, the effect of different metal ions on pectinase activity was also studied.

  2. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

    OpenAIRE

    S.M. Azwai; E.A. Alfallani; S.K. Abolghait; A.M. Garbaj; H.T. Naas; A.A. Moawad; F.T. Gammoudi; H.M. Rayes; I. Barbieri; I.M. Eldaghayes

    2016-01-01

    The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localitie...

  3. Aspergillus: introduction

    Science.gov (United States)

    Species in the genus Aspergillus possess versatile metabolic activities that impact our daily life both positively and negatively. Aspergillus flavus and Aspergillus oryzae are closely related fungi. While the former is able to produce carcinogenic aflatoxins and is an etiological agent of aspergill...

  4. Occurrence and Variety of β-Lactamase Genes among Aeromonas spp. Isolated from Urban Wastewater Treatment Plant

    Directory of Open Access Journals (Sweden)

    Marta Piotrowska

    2017-05-01

    Full Text Available Members of the genus Aeromonas that commonly occur in various aquatic ecosystems are taken into account as vectors spreading antibiotic resistance genes (ARGs in the environment. In our study strains of Aeromonas spp. (n = 104 not susceptible to ampicillin were isolated from municipal sewage of different levels of purification – raw sewage, activated sludge and treated wastewater. The crucial step of the study was the identification of β-lactamase resistance genes. The identified genes encode β-lactamases from 14 families – blaTEM, blaOXA, blaSHV, blaCTX-M, blaMOX, blaACC, blaFOX, blaGES, blaPER, blaV EB, blaKPC, cphA, imiH, and cepH. There were no significant differences in number of identified ARGs between isolation points. BlaOXA, blaFOX variants and, characteristic for Aeromonas genus, metallo-β-lactamase cphA-related genes were the most commonly identified types of β-lactam resistance determinants. Moreover, we found four extended-spectrum β-lactamases (blaSHV -11, blaCTX-M-27, blaCTX-M-98, and blaPER-4 – and seven AmpC (blaACC, blaFOX-2-like, blaFOX-3, blaFOX-4-like, blaFOX-9, blaFOX-10-like, and blaFOX-13-like types and variants of genes that had never been found among Aeromonas spp. before. Five of the β-lactamases families (blaTEM, blaOXA, blaFOX, blaV EB, and cphA were identified in all three isolation sites, which supports the hypothesis that wastewater treatment plants (WWTPs are hot spots of ARGs dissemination. The obtained ARGs sequences share high identity with previously described β-lactamases, but new variants of those genes have to be considered as well. Characterization of antibiotic susceptibility was performed using disk the diffusion method with 12 different antibiotics according to CLSI guidelines. Over 60% of the strains are unsusceptible to cefepime and chloramphenicol and the majority of the strains have a multidrug resistance phenotype (68%. Finally, analysis of plasmid profiles among the resistant strains

  5. Diverse and Abundant Secondary Metabolism Biosynthetic Gene Clusters in the Genomes of Marine Sponge Derived Streptomyces spp. Isolates

    Directory of Open Access Journals (Sweden)

    Stephen A. Jackson

    2018-02-01

    Full Text Available The genus Streptomyces produces secondary metabolic compounds that are rich in biological activity. Many of these compounds are genetically encoded by large secondary metabolism biosynthetic gene clusters (smBGCs such as polyketide synthases (PKS and non-ribosomal peptide synthetases (NRPS which are modular and can be highly repetitive. Due to the repeats, these gene clusters can be difficult to resolve using short read next generation datasets and are often quite poorly predicted using standard approaches. We have sequenced the genomes of 13 Streptomyces spp. strains isolated from shallow water and deep-sea sponges that display antimicrobial activities against a number of clinically relevant bacterial and yeast species. Draft genomes have been assembled and smBGCs have been identified using the antiSMASH (antibiotics and Secondary Metabolite Analysis Shell web platform. We have compared the smBGCs amongst strains in the search for novel sequences conferring the potential to produce novel bioactive secondary metabolites. The strains in this study recruit to four distinct clades within the genus Streptomyces. The marine strains host abundant smBGCs which encode polyketides, NRPS, siderophores, bacteriocins and lantipeptides. The deep-sea strains appear to be enriched with gene clusters encoding NRPS. Marine adaptations are evident in the sponge-derived strains which are enriched for genes involved in the biosynthesis and transport of compatible solutes and for heat-shock proteins. Streptomyces spp. from marine environments are a promising source of novel bioactive secondary metabolites as the abundance and diversity of smBGCs show high degrees of novelty. Sponge derived Streptomyces spp. isolates appear to display genomic adaptations to marine living when compared to terrestrial strains.

  6. A comparison of three methods for the isolation of Arcobacter spp. from retail raw poultry in Northern Ireland

    DEFF Research Database (Denmark)

    Scullion, R.; Harrington, C. S.; Madden, R. H.

    2004-01-01

    Recent evidence suggests that arcobacters, especially Arcobacter butzleri, are potential foodborne pathogens, but standardized detection methods have yet to be established. A study was undertaken to determine which of three isolation methods was the most effective for the isolation of Arcobacter...... spp. from fresh raw poultry. Method I was microaerobic and involved a membrane filtration step followed by plating onto blood agar. Method 2 was also microaerobic and involved enrichment and plating media containing a five-antibiotic cocktail. Method 3 was aerobic and was based on enrichment...... in a charcoal-based broth containing two antibiotics. Retail poultry samples (n = 50) were obtained from supermarkets in Northern Ireland; the European Community license number was recorded to ensure sample diversity. Presumptive arcobacters were identified using genus-specific and species-specific primers...

  7. Free chlorine and monochloramine inactivation kinetics of Aspergillus and Penicillium in drinking water.

    Science.gov (United States)

    Ma, Xiao; Bibby, Kyle

    2017-09-01

    Fungi are near-ubiquitous in potable water distribution systems, but the disinfection kinetics of commonly identified fungi are poorly studied. In the present study, laboratory scale experiments were conducted to evaluate the inactivation kinetics of Aspergillus fumigatus, Aspergillus versicolor, and Penicillium purpurogenum by free chlorine and monochloramine. The observed inactivation data were then fit to a delayed Chick-Watson model. Based on the model parameter estimation, the Ct values (integrated product of disinfectant concentration C and contact time t over defined time intervals) for 99.9% inactivation of the tested fungal strains ranged from 48.99 mg min/L to 194.7 mg min/L for free chlorine and from 90.33 mg min/L to 531.3 mg min/L for monochloramine. Fungal isolates from a drinking water system (Aspergillus versicolor and Penicillium purpurogenum) were more disinfection resistant than Aspergillus fumigatus type and clinical isolates. The required 99.9% inactivation Ct values for the tested fungal strains are higher than E. coli, a commonly monitored indicator bacteria, and within a similar range for bacteria commonly identified within water distribution systems, such as Mycobacterium spp. and Legionella spp. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Occurrence of emerging food-borne pathogenic Arcobacter spp. isolated from pre-cut (ready-to-eat) vegetables.

    Science.gov (United States)

    Mottola, Anna; Bonerba, Elisabetta; Bozzo, Giancarlo; Marchetti, Patrizia; Celano, Gaetano Vitale; Colao, Valeriana; Terio, Valentina; Tantillo, Giuseppina; Figueras, Maria José; Di Pinto, Angela

    2016-11-07

    Given that changes in consumer food behaviours have led to an increase in the demand for pre-cut ready-to-eat (RTE) vegetables, and that few data are currently available on the occurrence of Arcobacter spp. in such foods, the aim of the present study was to assess the occurrence of Arcobacter spp. that carry virulence-associated genes on pre-cut RTE vegetables, using cultural and molecular methods. Arcobacter was detected using biomolecular identification methods in 44/160 (27.5%) of the samples, of which 40/44 (90.9%) isolates corresponded to A. butzleri and 4/44 (9.1%) to A. cryaerophilus. Studying the incidence of 9 virulence-associated genes revealed the widespread distribution of these genes among the Arcobacter isolates tested. The results obtained in our research provided plenty of information on the health risks associated with the direct consumption of raw vegetables, and highlight the need to implement further studies at each level of the production chain, in order to obtain further information to help protect human health. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. The effects of inulin and fructo-oligosaccharide on the probiotic properties of Lactobacillus spp. isolated from human milk.

    Science.gov (United States)

    Tulumoğlu, Şener; Erdem, Belgin; Şimşek, Ömer

    2018-05-22

    This study aims to determine the effects of inulin and fructo-oligosaccharide (FOS) on the probiotic properties of five Lactobacillus spp. isolated from human milk. Lactobacillus spp. were isolated and identified, and the growth characteristics, acid and bile salt tolerance, antagonistic effects, and cholesterol assimilation of Lactobacillus strains were investigated in the presence of inulin and FOS. Lactobacillus casei L1 was able to utilize inulin and FOS as carbon source as well as glucose even other strains were able to use, including Lactobacillus rhamnosus GG. This strain also showed high tolerance to acid and bile salt, even at pH 2.5 and 0.5% bile salt levels, respectively. Inulin and FOS promoted the antimicrobial activity of L. casei L1 against pathogenic bacteria. Cholesterol assimilation was higher than in the other and control probiotic strains in the presence inulin and FOS, which were measured as 14 and 25 mg/dL, respectively. In conclusion, L. casei L1 can use both inulin and FOS to maintain its viability both at digestive conditions and also the relevant prebiotics, and show broad antagonistic activity and cholesterol assimilation.

  10. Study of strains of Candida spp. Isolated from catheters in UHC of Oran (Algeria): Identification and antifungal susceptibility.

    Science.gov (United States)

    Bendjelloul, M; Boucherit-Otmani, Z; Boucherit, K

    2016-09-01

    The increasing incidence of Candida spp., and the vital prognosis often compromise for patients with Candida species make urgent the exact knowledge of their distribution worldwide and exhaust action antifungals currently used in clinical. That why we carry out an epidemiological study of Candida species and testing their susceptibility against two antifungals: amphotericin B and caspofungin. Samplings of peripheral venous catheters (PVC) were carried out from during 8months on the services of Internal medicine, Surgery A and Neonatology of Oran's University Hospital Center (UHC). The study of the susceptibility of Candida species to antifungal agents was performed according to the Clinical Laboratory Standards Institute (CLSI 2008). From 300 samples, 25 yeasts were isolated. The rate of colonization PVC was 8.33% by Candida spp. The most isolated strains were Candida parapsilosis with 64% of cases, followed by Candida albicans (12%) then 8% for Candida glabrata and Candida krusei. However, only 4% of isolates were Candida famata or Candida lusitaniae. Furthermore all isolated strains were susceptible to amphotericin B with Minimum Inhibitory Concentrations (MIC) ranging from 0.25 to 1μg/mL. MIC obtained with caspofungin vary from 0.0625 to 2μg/mL for all strains. Moreover, one strain of C. krusei is resistant to caspofungin with a MIC superior to 8μg/mL. All though caspofungin is at least as effective as amphotericin B, it is better tolerated for the treatment of invasive fungal infections. Copyright © 2016. Published by Elsevier Masson SAS.

  11. Mycotoxin producing potential of some isolates of Aspergillus flavus and Eurotium groups from meat products.

    Science.gov (United States)

    el-Kady, I; el-Maraghy, S; Zohri, A N

    1994-09-01

    All strains (92) of A. flavus group proved to be positive for production of aflatoxin (45 to 1200 micrograms/50 ml medium) on potato dextrose liquid medium, while 59 strains only proved to be positive (35-310 micrograms/50 ml) on 15% NaCl potato-dextrose liquid medium. Most of the strains tested of A. flavus, A. flavus var. columnaris and A. oryzae produced aflatoxins B1, B2, G1 & G2. All positive strains of A. tamarii produced aflatoxins G1 & G2 while the tested isolate of A. zonatus produced aflatoxins B1 & G1. Of 95 strains tested of Eurotium, aflatoxins B1 & G1 were produced by one strain of each of E. chevalieri var. intermedium, E. repens and E. rubrum. Gliotoxin was detected in the extract of two strains of E. chevalieri and one strain of each of E. chevalieri var. intermedium and E. pseudoglaucum on the salt-free medium, and two strains of each of E. chevalieri, E. chevalieri var. intermedium and one of E. pseudoglaucum on 15% NaCl medium. Sterigmatocystin was produced by some strains of E. chevalieri, E. chevalieri var. intermedium, E. amstelodami, E. pseudoglaucum and E. rubrum on the two experimental media. One strain only of E. repens produced ochratoxin A while citrinin was detected in the extract of one strain of E. pseudoglaucum.

  12. Molecular and biochemical characterization of Iranian surfactin-producing Bacillus subtilis isolates and evaluation of their biocontrol potential against Aspergillus flavus and Colletotrichum gloeosporioides.

    Science.gov (United States)

    Mohammadipour, Matin; Mousivand, Maryam; Salehi Jouzani, Gholamreza; Abbasalizadeh, Saeed

    2009-04-01

    The characterization of surfactin-producing Bacillus subtilis isolates collected from different ecological zones of Iran is presented. Characterization was performed using blood agar, PCR, drop-collapse, and reverse-phase high-performance liquid chromatography (HPLC) analyses, and the isolates' biocontrol effects against the aflatoxin-producing agent Aspergillus flavus and the citrus antracnosis agent Colletotrichum gloeosporioides were studied. In total, 290 B. subtilis isolates were isolated from phylosphere and rhizosphere samples collected from fields and gardens of 5 provinces of Iran. Blood agar assays showed that 185 isolates produced different biosurfactants. Isolates containing the sfp gene, coding for surfactin, were detected using the PCR method. It was found that 14 different isolates contained the sfp gene. Drop-collapse assays, which detect isolates with high production of surfactin, showed that 7 isolates produced high levels of surfactin. It was found from HPLC analysis that the isolates containin the sfp gene produced between 55 and 1610 mg of surfactin per litre of broth medium. Four isolates, named BS119m, BS116l, N3dn, and BS113c, produced more than 1000 mg of surfactin per litre of broth. The highest surfactin production level was observed for isolate BS119m (1610 mg/L). The antagonistic potential of the sfp gene-containing isolates was determined using dual culture and chloroform vapour methods. Our bioassay results indicated that isolate BS119m showed high inhibitory effects against A. flavus (100%) and C. gloeosporioides (88%). Furthermore, the effect of purified surfactin on the growth of A. flavus was evaluated. Mycelia growth was considerably reduced with increasing concentration of surfactin, and 36%, 54%, 84%, and 100% inhibitions of mycelia growth were, respectively, observed at 20, 40, 80, and 160 mg/L after 7 days of incubation.

  13. Isolation and Identification of Campylobacter spp. from Poultry and Poultry By-Products in Tunisia by Conventional Culture Method and Multiplex Real-Time PCR.

    Science.gov (United States)

    Jribi, Hela; Sellami, Hanen; Mariam, Siala; Smaoui, Salma; Ghorbel, Asma; Hachicha, Salma; Benejat, Lucie; Messadi-Akrout, Feriel; Mégraud, Francis; Gdoura, Radhouane

    2017-10-01

    Thermophilic Campylobacter spp. are one of the primary causes of bacterial human diarrhea. The consumption of poultry meats, by-products, or both is suspected to be a major cause of human campylobacteriosis. The aims of this study were to determine the prevalence of thermophilic Campylobacter spp. in fresh poultry meat and poultry by-products by conventional culture methods and to confirm Campylobacter jejuni and Campylobacter coli isolates by using the multiplex PCR assay. Two hundred fifty fresh poultry samples were collected from a variety of supermarkets and slaughterhouses located in Sfax, Tunisia, including chicken (n =149) and turkey (n =101). The samples were analyzed using conventional microbiological examinations according to the 2006 International Organization for Standardization method (ISO 10272-1) for Campylobacter spp. Concurrently, a real-time PCR was used for identification of C. jejuni and C. coli . Of the 250 samples of poultry meat and poultry by-products, 25.6% (n = 64) were contaminated with Campylobacter spp. The highest prevalence of Campylobacter spp. was found in chicken meat (26.8%) followed by turkey meat (23.7%). Among the different products, poultry breasts showed the highest contamination (36.6%) followed by poultry by-products (30%), poultry wings (28%) and poultry legs (26%) showed the lowest contamination, and no contamination was found on neck skin. Of the 64 thermophilic Campylobacter isolates, C. jejuni (59.7%) was the most frequently isolated species and 10.9% of the isolates were identified as C. coli . All of the 64 Campylobacter isolates identified by the conventional culture methods were further confirmed by PCR. The seasonal peak of Campylobacter spp. contamination was in the warm seasons (spring and summer). The study concluded that high proportions of poultry meat and poultry by-products marketed in Tunisia are contaminated by Campylobacter spp. Furthermore, to ensure food safety, poultry meats must be properly cooked

  14. Identification of ochratoxin A producing Aspergillus carbonarius and A. niger clade isolated from grapes using the loop-mediated isothermal amplification (LAMP) reaction.

    Science.gov (United States)

    Storari, M; von Rohr, R; Pertot, I; Gessler, C; Broggini, G A L

    2013-04-01

    To develop two assays based on the loop-mediated isothermal amplification (LAMP) of DNA for the quick and specific identification of Aspergillus carbonarius and ochratoxigenic strains of the Aspergillus niger clade isolated from grapes. Two sets of primers were designed based on the polyketide synthase genes involved or putatively involved in ochratoxin A (OTA) biosynthesis in A. carbonarius and A. niger clade. Hydroxynaphthol blue was used as indirect method to indicate DNA amplification. The limit of detection of both assays was comparable to that of a PCR reaction. Specificities of the reactions were tested using DNA from different black aspergilli isolated from grapes. The two LAMP assays were then used to identify A. carbonarius and ochratoxigenic A. niger and A. awamori grown in pure cultures without a prior DNA extraction. The two LAMP assays permitted to quickly and specifically identify DNA from OTA-producing black aspergilli, as well as isolates grown in pure culture. Monitoring vineyards for the presence of OTA-producing strains is part of the measures to minimize the occurrence of OTA in grape products. The two LAMP assays developed here could be potentially used to speed the screening process of vineyards for the presence of OTA-producing black aspergilli. © 2013 The Society for Applied Microbiology.

  15. Biodiversity and ITS-RFLP Characterisation of Aspergillus Section Nigri Isolates in Grapes from Four Traditional Grape-Producing Areas in Greece

    Science.gov (United States)

    Kizis, Dimosthenis; Natskoulis, Pantelis; Nychas, George-John E.; Panagou, Efstathios Z.

    2014-01-01

    A study on the occurrence of Aspergillus section Nigri species on grapes from four traditional grape-producing areas in Greece during the 2011/2012 vintage, and their capability to produce OTA was conducted. One hundred and twenty-eight black aspergilli isolates were characterised at the species level initially by the use of morphological criteria in accordance with appropriate keys, followed by molecular characterisation performed with Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR-RFLP) of the 5.8 ribosomal RNA gene Internal Transcribed Spacer region (5.8 rRNA ITS). Restriction enzyme digestion of the ITS amplicons using the HhaI, HinfI and RsaI, endonucleases distinguished eleven different patterns of restriction fragment length polymorphism (RFLP), four for each of the HhaI and RsaI digests and three for HinfI. From a total number of 128 individual isolates, 124 were classified into four Aspergillus species corresponding to A. carbonarius, A. tubingensis, A. japonicus and A. ibericus, and the remaining 4 were classified as members of the A. niger aggregate. A. carbonarius and A. tubingensis being the main representative species were equally counted, with higher geographical representation of the former in southern and the latter in northern regions, respectively. All isolates were tested for their ochratoxigenic potential by use of High Performance Liquid Chromatography (HPLC) and Enzyme Linked Immuno Sorbent Assay (ELISA), resulting in significant interspecies differences in OTA production. PMID:24710283

  16. Trends in Resistance to Extended-Spectrum Cephalosporins and Carbapenems among Escherichia coli and Klebsiella spp. Isolates in a District in Western India during 2004–2014

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    Ingvild Odsbu

    2018-01-01

    Full Text Available Surveillance data on the level of resistant bacteria is needed to inform strategies to reduce the development and spread of antibiotic resistance. The aim of this study was to determine the non-susceptibility trends to extended-spectrum cephalosporins and carbapenems among Escherichia coli and Klebsiella spp. isolates from the district of Nashik in Western India during the period 2004–2014. Antibacterial susceptibility testing of clinical isolates was performed using Kirby-Bauer disc diffusion method to determine inhibitory zone diameters. The change in proportions of non-susceptible bacteria over calendar time was investigated with spline transformations in a logistic regression model. For the extended-spectrum cephalosporins, the proportions of non-susceptible E. coli and Klebsiella spp. isolates were above 78.4% and 84.9% throughout the study period, respectively. E. coli and Klebsiella spp. isolates exhibited carbapenem non-susceptibility levels as high as 76.9% and 84.1% respectively. The proportions of extended-spectrum betalactamase (ESBL-producing isolates ranged from 38.3–85.9% in E. coli and from 45.1–93.1% in Klebsiella spp. Significantly higher proportions of non-susceptible and ESBL-producing isolates were found among isolates from inpatients compared to isolates from outpatients for both E. coli and Klebsiella spp. (p < 0.050. The high proportions of non-susceptible isolates observed show that there is great need to focus on optimal use of antibiotics to reduce the development of antibiotic resistance.

  17. Aspergillus asper sp. nov. and Aspergillus collinsii sp. nov., from Aspergillus section Usti.

    Science.gov (United States)

    Jurjevic, Zeljko; Peterson, Stephen W

    2016-07-01

    In sampling fungi from the built environment, two isolates that could not confidently be placed in described species were encountered. Phenotypic analysis suggested that they belonged in Aspergillus sect. Usti. In order to verify the sectional placement and to assure that they were undescribed rather than phenotypically aberrant isolates, DNA was isolated and sequenced at the beta-tubulin, calmodulin, internal transcribed spacer and RNA polymerase II loci and sequences compared with those from other species in the genus Aspergillus. At each locus, each new isolate was distant from existing species. Phylogenetic trees calculated from these data and GenBank data for species of the section Usti excluded the placement of these isolates in existing species, with statistical support. Because they were excluded from existing taxa, the distinct species Aspergillus asper (type strain NRRL 35910 T ) and Aspergillus collinsii (type strain NRRL 66196 T ) in sect. Usti are proposed to accommodate these strains.

  18. Molecular characterization of Ambler class A to D β-lactamases, ISAba1, and integrons reveals multidrug-resistant Acinetobacter spp. isolates in northeastern China.

    Science.gov (United States)

    Sun, Xiaoyu; Liu, Bin; Chen, Yan; Huang, Honglan; Wang, Guoqing; Li, Fan; Ni, Zhaohui

    2016-12-01

    The prevalence of various Ambler class A to D β-lactamases, ISAba1, and class 1 and 2 integrons as well as the clonal relatedness in 105 Acinetobacter spp. isolates found in northeastern China was investigated. All 105 Acinetobacter spp. isolates were determined to be multidrug resistant (MDR), and the resistance rates to carbapenem agents were approximately 50%. PER, IMP, AmpC, and OXA-23 were found to be dominant β-lactamases belonging to different classes, respectively. This is the first report of the coexistence of bla PER , bla IMP , bla AmpC , and bla OXA-23-like genes in Acinetobacter spp. isolates from northeastern China. ISAba1 was found upstream of the bla OXA-23-like gene in 87.8% (36/41) strains and upstream of the bla OXA-51-like gene in 26.5% (13/49) strains. ISAba3-like element was found upstream of the bla OXA-58-like gene in one bla OXA-58-like -positive strain. The presence of IntI1 was detected in 63.8% (67/105) of the isolates and the most prevalent gene cassettes were aacA4, aadA1, and catB8. The highly prevalent isolates belong to international clonal lineage (ICL)-II. These results indicate that the wide horizontal and clonal spread of MDR Acinetobacter spp. isolates harbouring multiple β-lactamase genes has become a serious problem in northeastern China.

  19. The potential of soil fungi associated with potato rhizosphere to control root knot nematode (Meloidogyne spp.) on potato

    Science.gov (United States)

    Utari, E.; Lisnawita; Safni, I.; Lubis, K.; Tantawi, AR; Hasanuddin

    2018-02-01

    The root knot nematode (Meloidogyne spp.) is one of important pathogens on potato crops in North Sumatra, Indonesia. This nematode causes significant crop losses on potatoes directly and indirectly. The effect of fungal isolates (Trichoderma sp. 1, Mucor sp.1, Aspergillus sp. 2, Mucor sp. 2) that were isolated from rhizosphere of potato in North Sumatra were studied in green house experiments on the growth of potato and the reproduction of the nematode (Meloidogyne spp). The results showed that Trichoderma sp. 1 caused a significant gall reduction, while Mucor sp.1 and Mucor sp.2 could improve the growth of potato.

  20. Prevalence, virulence factors and antifungal susceptibility of Candida spp. isolated from bloodstream infections in a tertiary care hospital in Brazil.

    Science.gov (United States)

    Canela, Heliara Maria Spina; Cardoso, Bárbara; Vitali, Lucia Helena; Coelho, Harnoldo Colares; Martinez, Roberto; Ferreira, Márcia Eliana da Silva

    2018-01-01

    Candida spp. are responsible for 80% of all systemic fungal infections and are associated with high mortality rates. This study characterised 79 bloodstream isolates of C. albicans, C. glabrata, C. orthopsilosis, C. parapsilosis and C. tropicalis from patients in a Brazilian hospital. The susceptibility to amphotericin B, caspofungin, fluconazole and voriconazole was determined; virulence factor production was assessed based on haemolysin, phospholipase and proteinase activities, and the patients' clinical characteristics were analysed. C. albicans was the predominant species (44%), followed by C. glabrata (19%), C. tropicalis (19%), C. parapsilosis (14%) and C. orthopsilosis (4%). The candidemia incidence was 1.52 per 1000 admissions, and the crude mortality rate was 52%. One C. albicans isolate was resistant to fluconazole and voriconazole. Moreover, 20.2%, 2.5% and 3.8% of the isolates exhibited dose-dependent susceptibility to fluconazole, voriconazole and caspofungin, respectively. In conclusion, although the C. glabrata incidence was higher than that usually described in Brazil, its increase was previously observed in studies conducted worldwide. Furthermore, the azole resistance of the C. albicans isolate could be due to previous exposure to these antifungals. These results highlight the importance of epidemiological studies and will facilitate an improved understanding of candidemia in the studied hospital. © 2017 Blackwell Verlag GmbH.

  1. Antimicrobial resistance in Escherichia coli and Salmonella spp. isolates from fresh produce and the impact to food safety.

    Science.gov (United States)

    Vital, Pierangeli G; Caballes, Marie Bernadine D; Rivera, Windell L

    2017-09-02

    Foodborne diseases associated with fresh produce consumption have escalated worldwide, causing microbial safety of produce of critical importance. Bacteria that have increasingly been detected in fresh produce are Escherichia coli and Salmonella spp., both of which have been shown to progressively display antimicrobial resistance. The study focused on the assessment of antimicrobial resistance of these enteric bacteria from different kinds of fresh produce from various open air markets and supermarkets in the Philippines. Using the disk diffusion assay on a total of 50 bacterial isolates obtained from 410 fresh produce surveyed, monoresistance to tetracycline was observed to be the most prevalent (38%), followed by multidrug resistance to tetracycline, chloramphenicol, ciprofloxacin, and nalidixic acid (4%), and lastly by dual resistance to tetracycline and chloramphenicol (2%). Using multiplex and simplex polymerase chain reaction (PCR) assays, tetA (75%) and tetB (9%) were found in tetracycline resistant isolates, whereas catI (67%) and catIII (33%) were detected in chloramphenicol resistant isolates. Sequence analysis of gyr and par genes from the ciprofloxacin and nalidixic acid resistant isolates revealed different mutations. Based on the results, fresh produce act as a reservoir of these antibiotic resistant bacteria which may pose health threat to consumers.

  2. Identification of clinical isolates of Aspergillus, including cryptic species, by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

    Science.gov (United States)

    Vidal-Acuña, M Reyes; Ruiz-Pérez de Pipaón, Maite; Torres-Sánchez, María José; Aznar, Javier

    2017-12-08

    An expanded library of matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been constructed using the spectra generated from 42 clinical isolates and 11 reference strains, including 23 different species from 8 sections (16 cryptic plus 7 noncryptic species). Out of a total of 379 strains of Aspergillus isolated from clinical samples, 179 strains were selected to be identified by sequencing of beta-tubulin or calmodulin genes. Protein spectra of 53 strains, cultured in liquid medium, were used to construct an in-house reference database in the MALDI-TOF MS. One hundred ninety strains (179 clinical isolates previously identified by sequencing and the 11 reference strains), cultured on solid medium, were blindy analyzed by the MALDI-TOF MS technology to validate the generated in-house reference database. A 100% correlation was obtained with both identification methods, gene sequencing and MALDI-TOF MS, and no discordant identification was obtained. The HUVR database provided species level (score of ≥2.0) identification in 165 isolates (86.84%) and for the remaining 25 (13.16%) a genus level identification (score between 1.7 and 2.0) was obtained. The routine MALDI-TOF MS analysis with the new database, was then challenged with 200 Aspergillus clinical isolates grown on solid medium in a prospective evaluation. A species identification was obtained in 191 strains (95.5%), and only nine strains (4.5%) could not be identified at the species level. Among the 200 strains, A. tubingensis was the only cryptic species identified. We demonstrated the feasibility and usefulness of the new HUVR database in MALDI-TOF MS by the use of a standardized procedure for the identification of Aspergillus clinical isolates, including cryptic species, grown either on solid or liquid media. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For

  3. Abundance, genetic diversity and sensitivity to demethylation inhibitor fungicides of Aspergillus fumigatus isolates from organic substrates with special emphasis on compost.

    Science.gov (United States)

    Santoro, Karin; Matić, Slavica; Gisi, Ulrich; Spadaro, Davide; Pugliese, Massimo; Gullino, Maria L

    2017-12-01

    Aspergillus fumigatus is a widespread fungus that colonizes dead organic substrates but it can also cause fatal human diseases. Aspergilloses are treated with demethylation inhibitor (DMI) fungicides; however, resistant isolates appeared recently in the medical and also environmental area. The present study aims at molecular characterizing and quantifying A. fumigatus in major environmental habitats and determining its sensitivity to medical and agricultural DMI fungicides. A. fumigatus was isolated only rarely from soil and meadow/forest organic matter but high concentrations (10 3 to 10 7  cfu/g) were detected in substrates subjected to elevated temperatures, such as compost and silage. High genetic diversity of A. fumigatus from compost was found based on SSR markers, distinguishing among fungal isolates even when coming from the same substrate sample, while subclustering was observed based on mutations in cyp51A gene. Several cyp51A amino acid substitutions were found in 15 isolates, although all isolates were fully sensitive to the tested DMI fungicides, with exception of one isolate in combination with one fungicide. This study suggests that the tested A. fumigatus isolates collected in Italy, Spain and Hungary from the fungus' major living habitats (compost) and commercial growing substrates are not potential carriers for DMI resistance in the environment. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  4. Obtención de ácido cítrico a partir de suero de leche por fermentación con Aspergillus spp.

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    Sánchez Toro Óscar Julián

    2004-07-01

    Full Text Available El suero de leche se ha constituido en el principal desecho de la industria láctea, a pesar de los constantes esfuerzos por aprovecharlo. El presente trabajo tuvo por objeto estudiar la obtención de ácido cítrico por fermentación sumergida con hongos del género Aspergillus, utilizando lactosuero en calidad de sustrato con miras a su aprovechamiento y a la reducción del impacto ambiental que causan los vertimientos de este subproducto en los cursos de agua. Se utilizaron las siguientes cepas: A. carbonarius NRRL 368, A. carbonarius NRRL 67 y A. niger NRRL 3. Fue seleccionado el mejor medio de adaptación para la propagación del inóculo. El diseño experimental planteado para evaluar la biosíntesis de ácido cítrico a partir de suero de leche modificado mediante diferentes tratamientos, dio como resultado que las dos cepas de A. carbonarius no presentaran diferencias significativas en la formación del ácido, mientras A. niger NRRL 3 alcanzó mayores concentraciones cuando se utilizó suero de leche desproteinizado, evaporado y con lactosa hidrolizada con f>-galactosidasa. A. carbonarius arrojó concentraciones promedio de ácido cítrico mayores que las encontradas para A. niger, lo que sugiere la necesidad de profundizar en su estudio como productor potencial. Se obtuvo la cinética de crecimiento celular, consumo de sustrato y formación del ácido en un biorreactor de tanque agitado con aireación de 3 L para el caso de A. niger, la cual fue simulada mediante modelos matemáticos no estructurados. Palabras clave: Aspergillus carbonarius, Aspergillus niger, biorreactor, simulación, p-galactosidasa.

  5. Antibiotic resistance monitoring in Vibrio spp. isolated from rearing environment and intestines of abalone Haliotis diversicolor.

    Science.gov (United States)

    Wang, R X; Wang, J Y; Sun, Y C; B L Yang; A L Wang

    2015-12-30

    546 Vibrio isolates from rearing seawater (292 strains) and intestines of abalone (254 strains) were tested to ten antibiotics using Kirby-Bauer diffusion method. Resistant rates of abalone-derived Vibrio isolates to chloramphenicol (C), enrofloxacin (ENX) and norfloxacin (NOR) were 40%) to kanamycin (KNA), furazolidone (F), tetracycline (TE), gentamicin (GM) and rifampin (RA). 332 isolates from seawater (n=258) and abalone (n=74) were resistant to more than three antibiotics. Peaked resistant rates of seawater-derived isolates to multiple antibiotics were overlapped in May and August. Statistical analysis showed that pH had an important effect on resistant rates of abalone-derived Vibrio isolates to RA, NOR, and ENX. Salinity and dissolved oxygen were negatively correlated with resistant rates of seawater-derived Vibrio isolates to KNA, RA, and PG. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Intracellular and membrane-damaging activities of methyl gallate isolated from Terminalia chebula against multidrug-resistant Shigella spp.

    Science.gov (United States)

    Acharyya, Saurabh; Sarkar, Prodipta; Saha, Dhira R; Patra, Amarendra; Ramamurthy, T; Bag, Prasanta K

    2015-08-01

    Shigella spp. (Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei) cause bacillary dysentery (shigellosis), which is characterized by bloody mucous diarrhoea. Although a variety of antibiotics have been effective for treatment of shigellosis, options are becoming limited due to globally emerging drug resistance. In the present study, in vitro antibacterial activity of methyl gallate (MG) isolated from Terminalia chebula was determined by performing MIC, minimal bactericidal concentration (MBC) and time-kill kinetic studies. Bacterial membrane-damaging activity of MG was determined by membrane perturbation and transmission electron microscopy (TEM). Cellular drug accumulation, cell infection and assessment of intracellular activities of MG and reference antibiotics were performed using HeLa cell cultures. The bactericidal activity of MG against multidrug-resistant (MDR) Shigella spp. in comparison with other commonly used drugs including fluoroquinolone was demonstrated here. TEM findings in the present study revealed that MG caused the total disintegration of inner and outer membranes, and leakage of the cytoplasmic contents of S. dysenteriae. The level of accumulation of MG and tetracycline in HeLa cells incubated for 24  h was relatively higher than that of ciprofloxacin and nalidixic acid (ratio of intracellular concentration/extracellular concentration of antibiotic for MG and tetracycline>ciprofloxacin and nalidixic acid). The viable number of intracellular S. dysenteriae was decreased in a time-dependent manner in the presence of MG (4 × MBC) and reduced to zero within 20  h. The significant intracellular activities of MG suggested that it could potentially be used as an effective antibacterial agent for the treatment of severe infections caused by MDR Shigella spp.

  7. Phenotypic and genotypic antimicrobial susceptibility pattern of Streptococcus spp. isolated from cases of clinical mastitis in dairy cattle in Poland.

    Science.gov (United States)

    Kaczorek, E; Małaczewska, J; Wójcik, R; Rękawek, W; Siwicki, A K

    2017-08-01

    Mastitis of dairy cattle is one of the most frequently diagnosed diseases worldwide. The main etiological agents of mastitis are bacteria of the genus Streptococcus spp., in which several antibiotic resistance mechanisms have been identified. However, detailed studies addressing this problem have not been conducted in northeastern Poland. Therefore, the aim of our study was to analyze, on phenotypic and genotypic levels, the antibiotic resistance pattern of Streptococcus spp. isolated from clinical cases of mastitis from dairy cattle in this region of Poland. The research was conducted using 135 strains of Streptococcus (Streptococcus uberis, n = 53; Streptococcus dysgalactiae, n = 41; Streptococcus agalactiae, n = 27; other streptococci, n = 14). The investigation of the antimicrobial susceptibility to 8 active substances applied in therapy in the analyzed region, as well as a selected bacteriocin (nisin), was performed using the minimum inhibitory concentration method. The presence of selected resistance genes (n = 14) was determined via PCR. We also investigated the correlation between the presence of resistance genes and the antimicrobial susceptibility of the examined strains in vitro. The highest observed resistance of Streptococcus spp. was toward gentamicin, kanamycin, and tetracycline, whereas the highest susceptibility occurred toward penicillin, enrofloxacin, and marbofloxacin. Additionally, the tested bacteriocin showed high efficacy. The presence of 13 analyzed resistance genes was observed in the examined strains [gene mef(A) was not detected]. In most strains, at least one resistance gene, mainly responsible for resistance to tetracyclines [tet(M), tet(K), tet(L)], was observed. However, a relationship between the presence of a given resistance gene and antimicrobial susceptibility on the phenotypic level was not always observed. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Proteome analysis of Aspergillus flavus isolate-specific responses to oxidative stress in relationship to aflatoxin production capability

    Science.gov (United States)

    Aspergillus flavus is an opportunistic pathogen that infects host plants such as maize and peanut under conducive conditions such as drought stress resulting in significant aflatoxin production. Drought-associated oxidative stress is known to exacerbate aflatoxin production by A. flavus. The object...

  9. IDENTIFICATION AND ANTIBIOTIC RESISTANCE PROFILE OFENTEROBACTERIACEAE SPECIES AND LACTOBACILLUS SPP. ISOLATED FROM HONEY BEES (APIS MELLIFERA DIGESTIVE TRACT

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2014-02-01

    Full Text Available Honey bees play important role in agricultural environment as main pollinators. Its important for many agricultural and wild plants. Also honey bee are producers of honey, which is consumed directly and it should be not a heat treatment. Many bacteria can be survive in honey for long time. Some of these bacteria are human and animal facultative pathogens, including Enterobactericaeae genera. If these bacteria contain antibiotic resistant genes than it can to leads to troubles in healing of some of bacterial infections. Lactobacillus spp. can be a reservoir of resistant genes for pathogenic bacterial strains. In this study we isolated Enterobacteriaceae strains from digestive tracts of honey bees. These strains was tested to the eight selected antibiotics by disc diffusion method and strains were indentified by MALDI TOF MS Biotyper. From this study we determined resistance to piperacillin in the highest level. Equally, we determined that Citrobacter gillenii was resistant to three antibiotics (piperacillin, chloramphenicol and levofloxacin from eight. Resistance to other antibiotics were determined in low levels and other indentified bacteria were resistant to one antibiotic, if any. Also we detected resistance in Lactobacillus spp. and determined MICs distribution for some selected antibiotics. For absence of similar studies we could not to discuss our results and we think that further experiments and studies are needed.

  10. Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready-to-eat salads, ham, and raw meat.

    Science.gov (United States)

    Pesavento, G; Calonico, C; Ducci, B; Magnanini, A; Lo Nostro, A

    2014-08-01

    Food specimens were analyzed in order to research Enterococcus spp.: 636 samples of raw meat (227 beef, 238 poultry, and 171 pork), 278 samples of cheese (110 fresh soft cheese and 168 mozzarella cheese), 214 samples of ready-to-eat salads, and 187 samples of ham. 312 strains of Enterococcus spp samples were isolated, then identified and submitted to susceptibility tests against 11 antimicrobial agents. The predominant species were Enterococcus faecalis in raw meat and Enterococcus faecium in retail products. Low percentages of microorganisms were resistant to vancomycin (3.53%), teicoplanin (2.24%), linezolid (0.32%), and amoxicillin in combination with clavulanic acid (0.32%). A high percentage of resistance was noted in E. faecalis at high level gentamicin (21.9%) and tetracycline (60.6%). In general, strains of E. faecalis were more resistant than E. faecium. Enterococci should be considered not only potential pathogens, but also a reservoir of genes encoding antibiotic resistance which can be transferred to other microorganisms. Continuous monitoring of their incidence and emerging resistance is important in order to identify foods which potentially represent a real risk to the population, and to ensure effective treatment of human enterococcal infections. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. L-Glutamic acid production by Bacillus spp. isolated from vegetable ...

    African Journals Online (AJOL)

    Ogiri” (fermented vegetable proteins) in Nigeria. The isolates were identified as Bacillus subtilis (6), (27.3%), Bacillus pumilus (5), (22.7%), Bacillus licheniformis (5), (27.3%) and Bacillus polymyxa (6), (22.7%). Four species of the Bacillus isolates ...

  12. Development of novel agar media for isolating guaiacol producing Alicyclobacillus spp.

    Science.gov (United States)

    Chang, S S; Park, S H; Kang, D H

    2013-06-03

    The purpose of this study is to develop a selective and differential medium (SK2 agar) for isolating guaiacol producing Alicyclobacillus. Forty-one selected dyes and vanillic acid were incorporated in SK agar for screening selective and differential agents. Two guaiacol producing (1016, 1101) and two non-guaiacol producing (19220, C-GD 1-1) Alicyclobacillus isolates were streaked onto media and color differentiation of the isolates was assessed. Among 41 tested dyes, Chrome Azurol S (CAS) allowed color differentiation of the two types of Alicyclobacillus. Colonies of guaiacol producing Alicyclobacillus isolates appeared as dark purple to royal blue color with yellow background, whereas non-guaiacol producing Alicyclobacillus isolates produced cream colored colonies with yellow background. Vanillic acid not only served as a precursor for guaiacol formation but also inhibited non-guaiacol producing Alicyclobacillus. Non-guaiacol producing isolates did not grow on SK agar containing more than 70 ppm vanillic acid, whereas the recovery of guaiacol producing isolates was unaffected. When compared with other Alicyclobacillus isolation media, not only was SK2 agar capable of selectively recovering guaiacol-producing Alicyclobacillus, the degree of growth was also approximately equal if not better than orange serum agar, potato dextrose agar, and K agar. The development of SK2 agar provides the fruit juice industry with an inexpensive, simple to use alternative for the detection of guaiacol producing Alicyclobacillus. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Isolation and identification of Helicobacter spp, from canine and feline gastric mucosa

    DEFF Research Database (Denmark)

    Jalava, K.; On, Stephen L.W.; VanDamme, P.A.R.

    1998-01-01

    It is known that virtually all healthy adult dogs and cats harbor spiral helicobacters in their gastric mucosa, Three species, Helicobacter felis, Helicobacter bizzozeronii, and Helicobacter salomonis have been isolated in vitro from the gastric mucosa of these animals. The aims of this study were...... conventional phenotypic tests, whole-cell protein profiling, and ultrastructural analysis in identifying the different species isolated from canine and feline gastric mucose. We cultured 95 and 22 gastric mucosal biopsies from dogs and cats, respectively. Twenty-one H. bizzozeronii strains, 8 H. felis strains......, 8 H. salomonis strains, 3 mixed cultures, 2 "Flexispira rappini"-like organisms, and 3 as get uncharacterized strains were isolated from the dogs, and 3 H. felis strains were isolated from the cats. The methods used here yielded Helicobacter isolation rates of 51% from dogs and 13.6% from cats...

  14. Prevalence and antibiotic-resistance characteristics of Enterococcus spp. Isolated from free-living and captive raptors in Central Illinois.

    Science.gov (United States)

    Marrow, Judilee; Whittington, Julia K; Mitchell, Mark; Hoyer, Lois L; Maddox, Carol

    2009-04-01

    Due to their predatory nature, raptor species may serve as important indicators of environmental contamination with antimicrobial-resistant bacteria. Raptors prey on small rodents and birds that have diverse habitat ranges, including urban and rural environments, and their intestinal microflora can reflect that of the animals on which they feed. Enterococcus spp. were selected as target organisms because they have been isolated from the avian gastrointestinal tract, can be conferred by prey items, and because they are capable of multiple resistance patterns. They are also a concerning source of human antimicrobial resistance. In this study fecal cultures were obtained from 15 May 2004 to 31 August 2004, from 21 free-living raptors and four captive raptors. Enterococcus was isolated from 21 (84%) of the 25 birds, and 54 isolates were chosen for further study based upon unique colony morphology. The most common isolate recovered was Enterococcus faecalis (95%, 95% confidence interval [CI]: 89-100). One bird in the study was determined to have Enterococcus gallinarum. Two distinct ribotypes of E. faecalis were identified, one with unique bands at 11 and 13 kb and the other with unique bands at 14 and 20 kb. Both ribotypes were found in free-living and captive birds. The Enterococcus isolates in this study demonstrated a variety of antimicrobial-resistance characteristics, including almost complete resistance to amikacin, first-generation cephalosporins, spectinomycin, and sulphadimethoxime. Isolates demonstrated variable resistance to chloramphenicol, gentamicin, enrofloxacin, erythromycin, and ticarcillin. No phenotypically vancomycin-resistant E. faecalis isolates were recovered from any of the raptors; three isolates had intermediate level susceptibility. A significantly higher number of isolates collected from captive birds demonstrated resistance to chloramphenicol than those obtained from free-living birds. This trend was not duplicated with any of the remaining

  15. Lactic acid bacteria as functional probiotic isolates for inhibiting the growth of Aspergillus flavus, A. parasiticus, A. niger and Penicillium chrysogenum.

    Science.gov (United States)

    Abbaszadeh, S; Tavakoli, R; Sharifzadeh, A; Shokri, H

    2015-12-01

    The aim of this study was to assess the potential of lactic acid bacteria (LAB) such as Lactobacillus acidophilus, L. rhamnosus, L. casei, L. paracasei and Bifidobacterium bifidum to inhibit the outgrowth of some common food-spoiling fungi including Aspergillus niger, A. flavus, A. parasiticus and Penicillium chrysogenum. Bacterial isolates were cultured on Mann Rogosa Sharpe (MRS) broth and liquid cultures and supernatants were prepared. The antifungal activity was tested using the agar well diffusion method. Both liquid culture and supernatant of L. casei isolate exhibited high antifungal activity, followed by L. acidophilus and L. paracasei isolates. The least activity was recorded for the isolates B. bifidum, while the isolate L. rhamnosus was moderately active against tested fungi. The antifungal activity of the supernatants obtained from all probiotic isolates against fungi was significantly less than that of liquid cultures (Pniger and A. parasiticus. These results suggest that probiotic bacteria strains have the ability to prevent the growth of pathogenic and mycotoxigenic fungi as antifungal agents for various biomedical applications. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  16. Characterization and Extracellular Enzyme Activity of Predominant Marine Bacillus spp. Isolated From Sea Water of Orissa Coast, India

    Directory of Open Access Journals (Sweden)

    Bal, S.

    2009-01-01

    Full Text Available Bacillus species are ubiquitous and diverse both in the terrestrial and marine ecosystems. In this investigation, predominant Bacillus species from sea water of three different sites of Orissa Coast were isolated and identified. In total, 16 Bacillus species were identified using morpho-physiological and biochemical characterisation. These identified bacterial strains include B. fastidiosus (CMB1, B. alvei (CMB2, B. coagulans (CMB3, B. marinus (CMB5, B. mycoides (CMB8, B. coagulans (PMB1, B. circulans (PMB2, B. cereus (PMB3, B. subtilis (PMB4, B. alcalophilus (GMB1, B. licheniformics (GMB2, B. polymyxa (GMB3 and B. pumilus (GMB4. The isolates CMB4, CMB6 and CMB7 were identified only up to genus level. These isolates were further screened for their salt tolerance and growth under varied temperature and pH conditions. Ability of these strains to produce extracellular enzymes such as protease, amylase, lipase, gelatinase, casein hydrolase, lecithinase, chitinase and pectinase were also screened and found that most of the Bacillus spp. possess extracellular enzymes.

  17. Assessment of antibiotic resistance of Escherichia coli isolates and screening of Salmonella spp. in wild ungulates from Portugal.

    Science.gov (United States)

    Dias, Diana; Torres, Rita T; Kronvall, Göran; Fonseca, Carlos; Mendo, Sónia; Caetano, Tânia

    2015-09-01

    Antibiotic resistance is an emerging global problem. Wild animals are rarely exposed to antibiotics and therefore low levels of antibiotic resistance are expected. However, the growing interactions of these animals with humans and livestock may have a huge impact on their bacterial flora. This study aimed to assess the levels of antibiotic resistance in Escherichia coli isolated from widespread wild ungulates in Portugal. The interpretation of inhibition zone diameters was performed according to clinical breakpoints and epidemiological cut-offs, determined with the normalized resistance interpretation (NRI) method. For clinical breakpoints, 16% of the isolates were resistant to at least one antibiotic, including ampicillin (10%), tetracycline (9%), streptomycin (5%) co-trimoxazole (4%), amoxicillin/clavulanic acid (1%) and cefoxitin (1%). The levels of resistance detected in E. coli strains isolated from wild boar were statistically different for ampicillin and co-trimoxasol. According to NRI cut-offs, 10% of the population showed a non-wild-type phenotype against at least one antibiotic, also including tetracycline (9%), co-trimoxazole (6%), streptomycin (4%), ampicillin (2%) and amoxicillin/clavulanic acid (1%). Considering this parameter of comparison, no statistically different levels of resistance were identified between E. coli recovered from the three wild ungulates. Screening of Salmonella spp., which can be potentially pathogenic, was also performed, revealing that its prevalence was very low (1.5%). The study demonstrated that wild ungulates from Portugal are also reservoirs of antibiotic-resistant bacteria. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  18. (+)-Geodin from Aspergillus terreus

    DEFF Research Database (Denmark)

    Rønnest, Mads Holger; Nielsen, Morten Thrane; Leber, Blanka

    2011-01-01

    The fungal metabolite (+)-geodin [systematic name: (2R)-methyl 5,7-dichloro-4-hydroxy-6'-methoxy-6-methyl-3,4'-dioxospiro[benzofuran-2,1'-cyclohexa-2',5'-diene]-2'-carboxylate], C(17)H(12)Cl(2)O(7), was isolated from Aspergillus terreus. The crystal structure contains two independent molecules...

  19. Variation in fumonisin and ochratoxin production associated with differences in biosynthetic gene content in Aspergillus niger and A. welwitschiae isolates from multiple crop and geographic origins

    Directory of Open Access Journals (Sweden)

    Antonia Susca

    2016-09-01

    Full Text Available The fungi Aspergillus niger and A. welwitschiae are morphologically indistinguishable species used for industrial fermentation and for food and beverage production. The fungi also occur widely on food crops. Concerns about their safety have arisen with the discovery that some isolates of both species produce fumonisin (FB and ochratoxin A (OTA mycotoxins. Here, we examined FB and OTA production as well as the presence of genes responsible for synthesis of the mycotoxins in a collection of 92 A. niger/A. welwitschiae isolates from multiple crop and geographic origins. The results indicate that i isolates of both species differed in ability to produce the mycotoxins; ii FB-nonproducing isolates of A. niger had an intact fumonisin biosynthetic gene (fum cluster; iii FB-nonproducing isolates of A. welwitschiae exhibited multiple patterns of fum gene deletion; and iv OTA-nonproducing isolates of both species lacked the ochratoxin A biosynthetic gene (ota cluster. Analysis of genome sequence data revealed a single pattern of ota gene deletion in the two species. Phylogenetic analysis suggest that the simplest explanation for this is that ota cluster deletion occurred in a common ancestor of A. niger and A. welwitschiae, and subsequently both the intact and deleted cluster were retained as alternate alleles during divergence of the ancestor into descendent species. Finally, comparison of results from this and previous studies indicate that a majority of A. niger isolates and a minority of A. welwitschiae isolates can produce FBs, whereas a minority of isolates of both species produce OTA. The comparison also suggested that the relative abundance of each species and frequency of FB/OTA-producing isolates can vary with crop and/or geographic origin.

  20. Isolation of Campylobacter spp. from Client-Owned Dogs and Cats, and Retail Raw Meat Pet Food in the Manawatu, New Zealand.

    Science.gov (United States)

    Bojanić, K; Midwinter, A C; Marshall, J C; Rogers, L E; Biggs, P J; Acke, E

    2017-09-01

    Campylobacter causes acute gastroenteritis in people worldwide and is frequently isolated from food, animals and the environment. The disease is predominately food-borne but many routes of transmission and sources of infection have been described, including contact with pets. The prevalence of Campylobacter spp. in dogs and cats varies widely, and data on New Zealand pets are limited. This study aimed to investigate the prevalence of Campylobacter spp. in dogs, cats and retail raw meat pet food products in New Zealand and to characterize Campylobacter jejuni isolates using multilocus sequence typing (MLST). Ninety dogs and 110 cats examined at the Massey University Veterinary Teaching Hospital for elective procedures, and fifty locally purchased retail raw meat pet diets were sampled. Two culture protocols combining Bolton broth enrichment and mCCDA and CAT agars in a microaerobic atmosphere at 42°C and 37°C with species identification using PCR were performed. The prevalence of Campylobacter spp., C. jejuni, Campylobacter upsaliensis and Campylobacter helveticus was 36%, 13%, 23% and 1% in dogs and 16%, 5%, 5% and 7% in cats, respectively. One dog had Campylobacter lari confirmed, and three dogs and one cat had multiple Campylobacter spp. detected. Significantly more animals tested positive using CAT than mCCDA agar (P dogs, whereas attendance for dental treatment was a risk factor for cats. Campylobacter spp. were isolated from 28%, C. jejuni 22%, C. lari 6% and Campylobacter coli 6% of food samples. Six isolates positive by Campylobacter genus PCR were identified as Arcobacter butzleri. Poultry meat was more likely to be positive than non-poultry meat (P = 0.006). Of the 13 C. jejuni pet isolates with full MLST profiles, eight were of different sequence types (ST) and all nine food isolates were of different STs. © 2016 Blackwell Verlag GmbH.

  1. In vitro activity of solithromycin and its metabolites, CEM-214 and N-acetyl-CEM-101, against 100 clinical Ureaplasma spp. isolates compared with azithromycin.

    Science.gov (United States)

    Furfaro, Lucy L; Spiller, O Brad; Keelan, Jeffrey A; Payne, Matthew S

    2015-09-01

    There is a strong association between vaginal and/or amniotic fluid Ureaplasma spp. colonisation and risk of preterm birth. The novel fluoroketolide antibiotic solithromycin (CEM-101) is active against Ureaplasma spp. in vitro. Evidence from ex vivo and in vivo models suggests that, unlike most macrolide antibiotics, solithromycin readily crosses the placenta. Solithromycin metabolism varies according to species; in pregnant sheep, the bioactive metabolites CEM-214 and N-acetyl-CEM-101 (NAc-CEM-101) have been shown to accumulate in the amniotic cavity following maternal solithromycin administration, potentially contributing to its antimicrobial effects. To determine the antimicrobial activity of these metabolites against Ureaplasma spp., the effects of solithromycin, CEM-214, NAc-CEM-101 and the comparator azithromycin were tested on a collection of 100 clinical Ureaplasma spp. isolates from the UK and Australia using a modified 96-well broth microdilution method. MIC90 values observed for the combined cohort were: solithromycin, 0.125 mg/L; CEM-214, 0.5mg/L; NAc-CEM-101, 0.5mg/L; and azithromycin, 2mg/L. Solithromycin showed 34-fold greater activity against Ureaplasma spp. isolates than azithromycin, whilst CEM-214 and NAc-CEM-101 possessed ca. 22% and 17% of the activity of solithromycin, respectively, significantly greater than that of azithromycin. One bacterial isolate showed resistance to azithromycin (MIC=16 mg/L) but had a much lower MIC for solithromycin (MIC=0.25mg/L). In conclusion, the metabolites of solithromycin had reduced, but still potent, activity against 100 clinical Ureaplasma spp. isolates in vitro. This may be important in some instances such as pregnancy, however studies to determine levels of the metabolites in these settings are required. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  2. Isolation and characterization of two mitoviruses and a putative alphapartitivirus from Fusarium spp.

    Science.gov (United States)

    Osaki, Hideki; Sasaki, Atsuko; Nomiyama, Koji; Sekiguchi, Hiroyuki; Tomioka, Keisuke; Takehara, Toshiaki

    2015-06-01

    The filamentous fungus Fusarium spp. includes several important plant pathogens. We attempted to reveal presence of double-stranded (ds) RNAs in the genus. Thirty-seven Fusarium spp. at the MAFF collection were analyzed. In the strains of Fusarium coeruleum, Fusarium globosum and Fusarium solani f. sp. pisi, single dsRNA bands were detected. The strains of F. coeruleum and F. solani f. sp. pisi cause potato dry rot and mulberry twig blight, respectively. Sequence analyses revealed that dsRNAs in F. coeruleum and F. globosum consisted of 2423 and 2414 bp, respectively. Using the fungal mitochondrial translation table, the positive strands of these cDNAs were found to contain single open reading frames with the potential to encode a protein of putative 757 and 717 amino acids (molecular mass 88.5 and 84.0 kDa, respectively), similar to RNA-dependent RNA polymerases of members of the genus Mitovirus. These dsRNAs in F. coeruleum and F. globosum were assigned to the genus Mitovirus (family Narnaviridae), and these two mitoviruses were designated as Fusarium coeruleum mitovirus 1 and Fusarium globosum mitovirus 1. On the other hand, a positive strand of cDNA (1950 bp) from dsRNA in F. solani f. sp. pisi contained an ORF potentially encoding a putative RdRp of 608 amino acids (72.0 kDa). The putative RdRp was shown to be related to those of members of the genus of Alphapartitivirus (family Partitiviridae). We coined the name Fusarium solani partitivirus 2 for dsRNA in F. solani f. sp. pisi.

  3. Comparative pathogenicity of Vibrio spp., Photobacterium damselae ssp. damselae and five isolates of Aeromonas salmonicida ssp. achromogenes in juvenile Atlantic halibut (Hippoglossus hippoglossus).

    Science.gov (United States)

    Bowden, T J; Bricknell, I R; Preziosi, B M

    2018-01-01

    Juvenile Atlantic halibut (~100 mg, Hippoglossus hippoglossus) were exposed to Vibrio proteolyticus, a Vibrio spp. isolate, Photobacterium damselae ssp. damselae and five different isolates of Aeromonas salmonicida ssp. achromogenes via an hour-long bath immersion to ascertain their variation in pathogenicity to this fish species. Results were analysed using Kaplan-Meier survival analysis. Analysis of the data from challenges using A. salmonicida ssp. achromogenes revealed three survival values of zero and a spread of values from 0 to 28.43. Challenges using a Vibrio spp isolate, V. proteolyticus and P. damselae resulted in Kaplan-Meier survival estimates of 31.21, 50.41 and 57.21, respectively. As all bacterial species tested could induce juvenile halibut mortalities, they must all be considered as potential pathogens. However, the degree of pathogenicity of A. salmonicida is isolate dependent. © 2017 John Wiley & Sons Ltd.

  4. Species Distribution and Prevalence of Putative Virulence Factors in Mesophilic Aeromonas spp. Isolated from Fresh Retail Sushi

    Directory of Open Access Journals (Sweden)

    Sunniva Hoel

    2017-05-01

    Full Text Available Aeromonas spp. are ubiquitous bacteria that have received increasing attention as human pathogens because of their widespread occurrence in food, especially seafood and vegetables. The aim of this work was to assess the species identity and phylogenetic relationship of 118 Aeromonas strains isolated from fresh retail sushi from three producers, and to characterize the isolates with respect to genetic and phenotypic virulence factors. We also evaluate the potential hazard associated with their presence in ready-to-eat seafood not subjected to heat treatment. Mesophilic Aeromonas salmonicida was most prevalent (74%, followed by A. bestiarum (9%, A. dhakensis (5%, A. caviae (5%, A. media (4%, A. hydrophila (2%, and A. piscicola (1%. All isolates were considered potentially pathogenic due to the high prevalence of genes encoding hemolysin (hlyA (99%, aerolysin (aerA (98%, cytotoxic enterotoxin (act (86%, heat-labile cytotonic enterotoxin (alt (99%, and heat-stable cytotonic enterotoxin (ast (31%. The shiga-like toxins 1 and 2 (stx-1 and stx-2 were not detected. Moreover, there was heterogeneity in toxin gene distribution among the isolates, and the combination of act/alt/hlyA/aerA was most commonly detected (63%. β-hemolysis was species-dependent and observed in 91% of the isolates. All A. media and A. caviae strains were non-hemolytic. For isolates belonging to this group, lack of hemolysis was possibly related to the absence of the act gene. Swimming motility, linked to adhesion and host invasion, occurred in 65% of the isolates. Partial sequencing of the gyrB gene demonstrated its suitability as a genetic marker for Aeromonas species identification and for assessment of the phylogenetic relationship between the isolates. The gyrB sequence divergence within a given species ranged from 1.3 to 2.9%. A. bestiarum, A. salmonicida, and A. piscicola were the most closely related species; their sequences differed by 2.7–3.4%. The average gyrB sequence

  5. 16S Ribosomal DNA Characterization of Nitrogen-Fixing Bacteria Isolated from Banana (Musa spp.) and Pineapple (Ananas comosus (L.) Merril)

    Science.gov (United States)

    Magalhães Cruz, Leonardo; Maltempi de Souza, Emanuel; Weber, Olmar Baler; Baldani, José Ivo; Döbereiner, Johanna; de Oliveira Pedrosa, Fábio

    2001-01-01

    Nitrogen-fixing bacteria isolated from banana (Musa spp.) and pineapple (Ananas comosus (L.) Merril) were characterized by amplified 16S ribosomal DNA restriction analysis and 16S rRNA sequence analysis. Herbaspirillum seropedicae, Herbaspirillum rubrisubalbicans, Burkholderia brasilensis, and Burkholderia tropicalis were identified. Eight other types were placed in close proximity to these genera and other alpha and beta Proteobacteria. PMID:11319127

  6. Epidemiological trends and patterns of antimicrobial resistance of Shigella spp. isolated from stool cultures in two different populations in Southern Israel.

    Science.gov (United States)

    Peleg, Itai; Givon-Lavi, Noga; Leibovitz, Eugene; Broides, Arnon

    2014-03-01

    Southern Israel is inhabited by Bedouins, living in conditions similar to developing countries and Jews, living in conditions similar to developed countries. We determined the epidemiology of Shigella spp. in these populations. We retrospectively reviewed Shigella spp. stool isolations between 2005-2009. Overall, 3295 isolates were analyzed. S. sonnei was isolated in 2057/3295 (62.4%) and S. flexneri in 1058 (32.1%). S. sonnei was isolated in 1567/1707 (91.8%) from Jewish patients and S. flexneri in 931/1542 (60.4%) from Bedouin patients. Ampicillin resistance increased linearly from 217/373 (58.2%) in 2005 to 186/256 (72.7%) in 2009, (P Shigella spp. to ampicilin and trimethoprim-sulfamethoxazole were found in Jewish patients: 1527/1706 (89.5%) versus 977/1542 (63.4%) (P Shigella spp. infections can differ in populations residing in the same geographical area. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Specific biological properties of Listeria innocua spp. isolated in Primorye Territory

    Directory of Open Access Journals (Sweden)

    E. A. Zaitseva

    2017-01-01

    Full Text Available Rationale: Most cases of listeriosis are caused by the pathogenic Listeria monocytogenes. Some cases of isolation of L.  innocua with pathogenicity factors from foods have been published, as well as on the cases of the disease in humans caused by this species. Aim: To assess biological properties including potential pathogenicity of L.  innocua, isolated from food and environmental objects. Materials and methods: We performed microbiological study of L. innocua cultures isolated from foods (n = 35 and environmental objects (n = 15 on the territory of Primorye Territory (Russian Federation, as well as assessment of their sensitivity to antibiotics. Results: The studied L.  innocua cultures showed stable phenotypic features of their biological properties, such as morphology, typical colony growth on the medium with characteristic odor of fermented milk, blue or blue-green luminescence induced by inclined light, presence of catalase activity and absence of the oxidase activity. Only 38 ± 6.9% of L.  innocua demonstrated movements at T 22 °С. L.  innocua cultures did not ferment mannitol (100% of cultures; they degraded ramnose to its acid without gas (70 ± 6.5% and degraded xylose (42.8 ± 7%. Listeria isolated from vegetables and environmental objects could ferment ramnose (92.8 ± 7.2% of the studied cultures and xylose (28.5 ± 12.5% more frequently than L. innocua isolated from meat and fish foods. L.  innocua demonstrated variable biochemical activities towards mannose (92 ± 3.8%, saccharose (85.7 ± 7.8% and melesitose (76.2 ± 9.5%. L. innocua cultures with hemolytic activity (34 ± 6.7% (α or β  type were isolated, more commonly from fish products. All Listeria irrespective of their isolation source showed lipase activity. L.  innocua cultures from foods and environmental objects were highly sensitive to antimicrobials from the following classes: penicillins (ampicillin, carbenicillin, combined amoxicillin and clavulanic

  8. MOLECULAR IDENTIFICATION AND ANTIMICROBIAL RESISTANCE PATTERN OF SEVEN CLINICAL ISOLATES OF Nocardia spp. IN BRAZIL

    Directory of Open Access Journals (Sweden)

    Larissa Anuska Zeni CONDAS

    2015-06-01

    Full Text Available Nocardia is a ubiquitous microorganism related to pyogranulomatous infection, which is difficult to treat in humans and animals. The occurrence of the disease is on the rise in many countries due to an increase in immunosuppressive diseases and treatments. This report of cases from Brazil presents the genotypic characterization and the antimicrobial susceptibility pattern using the disk-diffusion method and inhibitory minimal concentration with E-test® strips. In summary, this report focuses on infections in young adult men, of which three cases were cutaneous, two pulmonary, one neurological and one systemic. The pulmonary, neurological and systemic cases were attributed to immunosuppressive diseases or treatments. Sequencing analysis of the 16S rRNA segments (1491 bp identified four isolates of Nocardia farcinica, two isolates of Nocardia nova and one isolate of Nocardia asiatica. N. farcinica was involved in two cutaneous, one systemic and other pulmonary cases; N. nova was involved in one neurological and one pulmonary case; and Nocardia asiatica in one cutaneous case. The disk-diffusion antimicrobial susceptibility test showed that the most effective antimicrobials were amikacin (100%, amoxicillin/clavulanate (100%, cephalexin (100% and ceftiofur (100%, while isolates had presented most resistance to gentamicin (43%, sulfamethoxazole/trimethoprim (43% and ampicillin (29%. However, on the inhibitory minimal concentration test (MIC test, only one of the four isolates of Nocardia farcinica was resistant to sulfamethoxazole/trimethoprim.

  9. Characterization of Salmonella spp. isolated from an integrated broiler chicken operation in Korea.

    Science.gov (United States)

    Lee, Young Ju; Kim, Hyun Jung; Park, Cheong Kyu; Kim, Ki Seuk; Bae, Dong Hwa; Kang, Min Su; Cho, Jae Keun; Kim, Ae Ran; Kim, Jong Wan; Kim, Byoung Han

    2007-04-01

    The purpose of this study was to investigate the biological and genetic characterization of persistent Salmonella isolates in an integrated broiler chicken operation, in an attempt to elucidate the source of contamination. From the breeder farm, the hatchery, the broiler farm and the chicken slaughter house of an integrated broiler chicken operation, a total of 6 serotypes were observed. Although S. Heidelberg was not detected in the broiler farm, it was consistently found in the breeder farm, the hatchery and the chicken slaughter house. Also, S. Enteritidis and S. Senftenberg were found in the hatchery and the chicken slaughter house, and the hatchery and the broiler farm, respectively. S. Gallinarum and S. Blockley were found only in the broiler farm, and S. Virchow was only recovered in the chicken slaughter house. Isolated S. Heidelberg, S. Enteritidis and S. Senftenberg strains were divided into 3, 5 and 7 types, respectively, on the basis of all properties. Especially, S. Senftenberg isolates, divided into four types by their antimicrobial resistance patterns, were all obviously the XbaI PFGE pattern. Also, four S. Enteritidis isolates resistant to nalidixic acid showed a difference in phage type and PFGE pattern. Such a different pattern was shown despite Salmonella isolates originating from an integrated broiler operation, suggesting that further epidemiological studies on many integrated chicken companies in Korea are needed.

  10. The production and activity test of cellulases using bagasse substrate on Aspergillus niger isolated from Clove field, Kare, Madiun

    Science.gov (United States)

    Ardhi, Muh. Waskito; Sulistyarsi, Ani; Pujiati

    2017-06-01

    Aspergillus sp is a microorganism which has a high ability to produce cellulase enzymes. In producing Cellulase enzymes requires appropriate concentration and incubation time to obtain optimum enzyme activity. This study aimed to determine the effect of inoculum concentration and incubation time towards production and activity of cellulases from Aspergillus sp substrate bagasse. This research used experiments method; completely randomized design with 2 factorial repeated 2 times. The treatment study include differences inoculum (K) 5% (K1), 15% (K2) 25%, (K3) and incubation time (F) that is 3 days (F1), 6 days (F2), 9 days (F3), 12 days (F4). The data taken from the treatment are glucose reduction and protein levels of crude cellulase enzyme activity that use Nelson Somogyi and Biuret methods. Analysis of variance ANOVA data used two paths with significance level of 5% then continued with LSD test. The results showed that: Fhit>Ftab. Thus, there is effect of inoculum concentrations and incubation time toward activity of crude cellulases of Aspergillus sp. The highest glucose reduction of treatment is K3F4 (concentration of inoculum is 25% with 12 days incubation time) amount 12.834 g / ml and the highest protein content is K3F4 (concentration of inoculum is 25% with with 12 days incubation time) amount 0.740 g / ml.

  11. A Brucella spp. Isolate from a Pac-Man Frog (Ceratophrys ornata) Reveals Characteristics Departing from Classical Brucellae.

    Science.gov (United States)

    Soler-Lloréns, Pedro F; Quance, Chris R; Lawhon, Sara D; Stuber, Tod P; Edwards, John F; Ficht, Thomas A; Robbe-Austerman, Suelee; O'Callaghan, David; Keriel, Anne

    2016-01-01

    Brucella are highly infectious bacterial pathogens responsible for brucellosis, a frequent worldwide zoonosis. The Brucella genus has recently expanded from 6 to 11 species, all of which were associated with mammals; The natural host range recently expanded to amphibians after some reports of atypical strains from frogs. Here we describe the first in depth phenotypic and genetic characterization of a Brucella strains isolated from a frog. Strain B13-0095 was isolated from a Pac-Man frog ( Ceratophyrus ornate ) at a veterinary hospital in Texas and was initially misidentified as Ochrobactrum anthropi . We found that B13-0095 belongs to a group of early-diverging brucellae that includes Brucella inopinata strain BO1 and the B. inopinata -like strain BO2, with traits that depart significantly from those of the "classical" Brucella spp. Analysis of B13-0095 genome sequence revealed several specific features that suggest that this isolate represents an intermediate between a soil associated ancestor and the host adapted "classical" species. Like strain BO2, B13-0095 does not possess the genes required to produce the perosamine based LPS found in classical Brucella , but has a set of genes that could encode a rhamnose based O-antigen. Despite this, B13-0095 has a very fast intracellular replication rate in both epithelial cells and macrophages. Finally, another major finding in this study is the bacterial motility observed for strains B13-0095, BO1, and BO2, which is remarkable for this bacterial genus. This study thus highlights several novel characteristics in strains belonging to an emerging group within the Brucella genus. Accurate identification tools for such atypical Brucella isolates and careful evaluation of their zoonotic potential, are urgently required.

  12. Isolation of nucleotide binding site-leucine rich repeat and kinase resistance gene analogues from sugarcane (Saccharum spp.).

    Science.gov (United States)

    Glynn, Neil C; Comstock, Jack C; Sood, Sushma G; Dang, Phat M; Chaparro, Jose X

    2008-01-01

    Resistance gene analogues (RGAs) have been isolated from many crops and offer potential in breeding for disease resistance through marker-assisted selection, either as closely linked or as perfect markers. Many R-gene sequences contain kinase domains, and indeed kinase genes have been reported as being proximal to R-genes, making kinase analogues an additionally promising target. The first step towards utilizing RGAs as markers for disease resistance is isolation and characterization of the sequences. Sugarcane clone US01-1158 was identified as resistant to yellow leaf caused by the sugarcane yellow leaf virus (SCYLV) and moderately resistant to rust caused by Puccinia melanocephala Sydow & Sydow. Degenerate primers that had previously proved useful for isolating RGAs and kinase analogues in wheat and soybean were used to amplify DNA from sugarcane (Saccharum spp.) clone US-01-1158. Sequences generated from 1512 positive clones were assembled into 134 contigs of between two and 105 sequences. Comparison of the contig consensuses with the NCBI sequence database using BLASTx showed that 20 had sequence homology to nuclear binding site and leucine rich repeat (NBS-LRR) RGAs, and eight to kinase genes. Alignment of the deduced amino acid sequences with similar sequences from the NCBI database allowed the identification of several conserved domains. The alignment and resulting phenetic tree showed that many of the sequences had greater similarity to sequences from other species than to one another. The use of degenerate primers is a useful method for isolating novel sugarcane RGA and kinase gene analogues. Further studies are needed to evaluate the role of these genes in disease resistance.

  13. Identification and antimicrobial susceptibility patterns of Staphylococcus spp. isolated from canine chronic otitis externa

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    Silva N.

    2001-01-01

    Full Text Available Swab samples obtained from 96 dogs with chronic otitis externa were cultured for the isolation of Staphylococcus species. Of 57 staphylococcal strains, 41 (72% were coagulase-negative (CNS. The identification of staphylococci strains was made by standard procedures for the routine identification of staphylococci in clinical practice. S. sciuri was the most frequent species isolated (22.8% from chronic otitis externa in dogs followed by S. intermedius (12.3%, S. auricularis (10.5% and S. aureus (8.8%. Three (5.2% CNS strains could not be identified. Bacterial isolates were susceptible to enrofloxacin, gentamicin, cephalothin, chloramphenicol and neomycin. Resistance was most common to penicillin G, oxacillin and ampicillin.

  14. Antibiotic Properties of the endophytic Streptomyces Spp. Isolated from the Leaves of Myanmar Medicinal Plants

    International Nuclear Information System (INIS)

    Aye Pe; Mar Mar Nyein; Win Maung

    2002-02-01

    Three medicinal plants of Myanmar are selected in the study of endophytic microorganisms and are taxonomically classified and identified to be Sa-ba-lin (Cymbopogon citratus Stapf.), Shazaungtinga- neah (Euphorbia splendens Bojer. ex Hooker) and Ma-shaw (Sauropus grandifolius Pax. and Hoffm.). The screening of endophytic microorganisms is performed according to the ISP method (International Streptomyces Projects 1993). The morphological and physicochemical properties of isolated strains are studied and identified to be the Genus Streptomyces. The test of apparent antimicrobial activity of isolated Streptomyces is done on 18 strains of pathogenic bacteria. It is found that the isolated endophytic Sireptomyces showed the significant antibacterial activity on most of the test organisms. (author)

  15. Loktanella spp. Gb03 as an algicidal bacterium, isolated from the culture of Dinoflagellate Gambierdiscus belizeanus.

    Science.gov (United States)

    Bloh, Anmar Hameed; Usup, Gires; Ahmad, Asmat

    2016-02-01

    Bacteria associated with harmful algal blooms can play a crucial role in regulating algal blooms in the environment. This study aimed at isolating and identifying algicidal bacteria in Dinoflagellate culture and to determine the optimum growth requirement of the algicidal bacteria, Loktanella sp. Gb-03. The Dinoflagellate culture used in this study was supplied by Professor Gires Usup's Laboratory, School of Environmental and Natural Resources Sciences, Faculty of Science and Technology, University Kebangsaan Malaysia, Malaysia. The culture was used for the isolation of Loktanella sp., using biochemical tests, API 20 ONE kits. The fatty acid content of the isolates and the algicidal activity were further evaluated, and the phenotype was determined through the phylogenetic tree. Gram-negative, non-motile, non-spore-forming, short rod-shaped, aerobic bacteria (Gb01, Gb02, Gb03, Gb04, Gb05, and Gb06) were isolated from the Dinoflagellate culture. The colonies were pink in color, convex with a smooth surface and entire edge. The optimum growth temperature for the Loktanella sp. Gb03 isolate was determined to be 30°C, in 1% of NaCl and pH7. Phylogenetic analysis based on 16S rRNA gene sequences showed that the bacterium belonged to the genus Loktanella of the class Alphaproteobacteria and formed a tight cluster with the type strain of Loktanella pyoseonensis (97.0% sequence similarity). On the basis of phenotypic, phylogenetic data and genetic distinctiveness, strain Gb-03, were placed in the genus Loktanella as the type strain of species. Moreover, it has algicidal activity against seven toxic Dinoflagellate. The algicidal property of the isolated Loktanella is vital, especially where biological control is needed to mitigate algal bloom or targeted Dinoflagellates.

  16. Petroleum degradation by endophytic Streptomyces spp. isolated from plants grown in contaminated soil of southern Algeria.

    Science.gov (United States)

    Baoune, Hafida; Ould El Hadj-Khelil, Aminata; Pucci, Graciela; Sineli, Pedro; Loucif, Lotfi; Polti, Marta Alejandra

    2018-01-01

    Petroleum hydrocarbons are well known by their high toxicity and recalcitrant properties. Their increasing utilization around worldwide led to environmental contamination. Phytoremediation using plant-associated microbe is an interesting approach for petroleum degradation and actinobacteria have a great potential for that. For this purpose, our study aimed to isolate, characterize, and assess the ability of endophytic actinobacteria to degrade crude petroleum, as well as to produce plant growth promoting traits. Seventeen endophytic actinobacteria were isolated from roots of plants grown naturally in sandy contaminated soil. Among them, six isolates were selected on the basis of their tolerance to petroleum on solid minimal medium and characterized by 16S rDNA gene sequencing. All petroleum-tolerant isolates belonged to the Streptomyces genus. Determination by crude oil degradation by gas chromatorgraph-flame ionization detector revealed that five strains could use petroleum as sole carbon and energy source and the petroleum removal achieved up to 98% after 7 days of incubation. These isolates displayed an important role in the degradation of the n-alkanes (C 6 -C 30 ), aromatic and polycyclic aromatic hydrocarbons. All strains showed a wide range of plant growth promoting features such as siderophores, phosphate solubilization, 1-aminocyclopropane-1-carboxylate deaminase, nitrogen fixation and indole-3-acetic acid production as well as biosurfactant production. This is the first study highlighting the petroleum degradation ability and plant growth promoting attributes of endophytic Streptomyces. The finding suggests that the endophytic actinobacteria isolated are promising candidates for improving phytoremediation efficiency of petroleum contaminated soil. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Herpetomonas spp. isolated from tomato fruits (Lycopersicon esculentum) in southern Spain.

    Science.gov (United States)

    Marín, Clotilde; Fabre, Sandrine; Sánchez-Moreno, Manuel; Dollet, Michel

    2007-05-01

    A flagellate of the family Trypanosomatidae was isolated from fruits of Lycopersicon esculentum (tomato) in southeastern Spain. The isolate was successfully adapted to in vitro culture in monophasic media. The morphology showed the kinetoplast to be positioned towards the middle of the body, and the typical opistomastigote form characteristic of members of the genus Herpetomonas. Amplification of the mini-exon gene was negative, whilst for the 5S ribosomal rRNA gene the result was positive. The DNA sequence was obtained and its alignment with other trypasomatids, obtained using the BLAST algorithm, suggested it was closely related to Herpetomonas samuelpessoai.

  18. Evaluation of matrix-assisted laser desorption/ionization time-of-fight mass spectrometry for identification of 345 clinical isolates of Aspergillus species from 11 Korean hospitals: comparison with molecular identification.

    Science.gov (United States)

    Park, Ju Heon; Shin, Jong Hee; Choi, Min Ji; Choi, Jin Un; Park, Yeon-Joon; Jang, Sook Jin; Won, Eun Jeong; Kim, Soo Hyun; Kee, Seung Jung; Shin, Myung Geun; Suh, Soon Pal

    2017-01-01

    We evaluated the ability of the Filamentous Fungi Library 1.0 of the MALDI-TOF MS Biotyper system to identify 345 clinical Aspergillus isolates from 11 Korean hospitals. Compared with results of the internal transcribed spacer region sequencing, the frequencies of correct identification at the species-complex level were 94.5% and 98.8% with cutoff values of 2.0 and 1.7, respectively. Compared with results of β-tubulin gene sequencing, the frequencies of correct identification at the species level were 96.0% (cutoff 2.0) and 100% (cutoff 1.7) for 303 Aspergillus isolates of five common, non-cryptic species, but only 4.8% (cutoff 1.7) and 0% (cutoff 2.0) for 42 Aspergillus isolates of six cryptic species (identifiable by β-tubulin or calmodulin sequencing). These results show that the MALDI Biotyper using the Filamentous Fungi Library version 1.0 enables reliable identification of the majority of common clinical Aspergillus isolates, although the database should be expanded to facilitate identification of cryptic species. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

    Science.gov (United States)

    Azwai, S.M.; Alfallani, E.A.; Abolghait, S.K.; Garbaj, A.M.; Naas, H.T.; Moawad, A.A.; Gammoudi, F.T.; Rayes, H.M.; Barbieri, I.; Eldaghayes, I.M.

    2016-01-01

    The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6 % of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 ×104 CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 ×104 CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products. PMID:27004169

  20. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

    Directory of Open Access Journals (Sweden)

    S.M. Azwai

    2016-03-01

    Full Text Available The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk. Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6% yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS with culture characteristics of Vibrio spp. More than half (n=27 of processed seafood samples (n=46 yielded colonies on TCBS, while only 44.6% of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 х104 CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 х104 CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9% were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

  1. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya.

    Science.gov (United States)

    Azwai, S M; Alfallani, E A; Abolghait, S K; Garbaj, A M; Naas, H T; Moawad, A A; Gammoudi, F T; Rayes, H M; Barbieri, I; Eldaghayes, I M

    2016-01-01

    The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6 % of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 ×10(4) CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 ×10(4) CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

  2. Genotypic characterization and species identification of Fasciola spp. with implications regarding the isolates infecting goats in Vietnam.

    Science.gov (United States)

    Nguyen, Thanh Giang Thi; Van De, Nguyen; Vercruysse, Jozef; Dorny, Pierre; Le, Thanh Hoa

    2009-12-01

    Ribosomal RNA sequences (361 or 362bp) of the second internal transcribed spacer 2 (ITS-2) and a portion of mitochondrial cox1 (423bp) for Fasciola spp. obtained from specimens collected in indigenous and hybrid goats and sheep in Vietnam were characterized for genotypic status and hybridization/introgression. Alignment of 48 ITS-2 sequences (also those from goats and sheep in this study) indicates that F. gigantica and F. hepatica differ typically from each other at seven sites whereas one of these is a distinguishing deletion (T) at the 327th position in F. gigantica relative to F. hepatica. The isolates from the mountainous goats in the North of Vietnam (Yen Bai province) showed the ITS-2 composition relatively identical to that of F. hepatica. The ITS-2 sequences from populations of Fasciola isolates in goats had probably experienced introgression/hybridization as reported previously in other ruminants and humans. All Vietnamese goat-of-origin specimens had high pairwise percentage of mitochondrial cox1 sequences to F. gigantica (97-100%), and very low identity to F. hepatica (91-93%), suggesting their maternal linkage to be traced to F. gigantica. The presence of hybrid and/or introgressed populations of liver flukes bearing genetic material from both F. hepatica and F. gigantica in the goats/sheep in Vietnam, regardless of indigenous or imported hosts, appears to be the first demonstration from a tropical country.

  3. Cryptococcus spp. isolation from excreta of pigeons (Columba livia) in and around Monterrey, Mexico.

    Science.gov (United States)

    Canónico-González, Yolanda; Adame-Rodríguez, Juan Manuel; Mercado-Hernández, Roberto; Aréchiga-Carvajal, Elva Teresa

    2013-01-01

    The presence of Cryptococcus spp. has been reported in Mexico's capital city; however, to our knowledge there are no reports of its presence in the state of Nuevo León located in northeast Mexico. This is presumed to be because the hot and dry climate in this region does not favor cryptococcal proliferation. This study confirmed the presence of C. neoformans and C. albidus in 20% (10/50) of randomly selected fecal samples of pigeons (Columba livia) in the Monterrey metropolitan area. The presence of this yeast in the state of Nuevo León is proof of its adaptation to the typically hot climate of the area and is consistent with recent reviews of cryptococcosis cases in several local hospitals. The two species were identified and characterized through microbiological tests and molecular identification by DNA extraction and PCR amplification of highly conserved 18S ribosomal DNA using ITS1 and ITS2 as target regions. The PCR products were sequenced and compared with those reported in GenBank.

  4. Spermatogenic and Phylo-molecular Characterizations of Isolated Fasciola Spp. From Cattle, North West Iran.

    Science.gov (United States)

    Rouhani, Soheila; Raeghi, Saber; Spotin, Adel

    2017-01-01

    Fascioliasis is economically important to the livestock industry that caused with Fasciola hepatica and Fasciola gigantica. The objective of this study was to identify these two species F. hepatica and F. gigantica by using nuclear and mitochondrial markers (ITS1, ND1 and CO1) and have been employed to analyze intraspecific phylogenetic relations of Fasciola spp. Approximately 150 Fasciola specimens were collected, then stained with haematoxylin-carmine dye and observed under an optical microscope to examine for the existence of sperm. The ITS1 marker was used to identify different Fasciola and phylogenetic analysis based on ND1 and CO1 sequence data were conducted by maximum likelihood algorithm. Fasciola samples were separated into 2 groups. Almost all specimens had many sperms in the seminal vesicle (spermic fluke) and one fluke did not contain any sperm in the seminal vesicle. The aspermic sample had F. gigantica RFLP pattern with ITS1 gene. Phylogenetic analysis based on NDI and COI sequence data were conducted by maximum likelihood showed a similar topology of the trees obtained particularly for F. hepatica and F. gigantica. This study demonstrated that aspermic Fasciola found in this region of Iran has same genetic structures through the spermic F. gigantica populations in accordance to phylogenetic tree.

  5. Activity of essential oils from spices against Staphylococcus spp. isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    M Dal Pozzo

    2011-10-01

    Full Text Available Avaliou-se a atividade antimicrobiana dos óleos essenciais (OE de Origanum vulgare (orégano, Thymus vulgaris (tomilho, Lippia graveolens (lipia, Zingiber officinale (gengibre, Salvia officinalis (sálvia, Rosmarinus officinalis (alecrim e Ocimum basilicum (manjericão, e de suas frações majoritárias, carvacrol e timol, frente a 32 isolados de Staphylococcus spp, oriundos de rebanhos leiteiros bovinos. A concentração inibitória mínima (CIM e a concentração bactericida mínima foram determinadas por meio da técnica de microdiluição em caldo. Orégano, tomilho e lípia (Orégano Mexicano apresentaram atividade antimicrobiana similar, médias geométrica de CIM de 1600µg mL-1; 1564µg mL-1; 1562µg mL-1, respectivamente, no entanto menos ativos que carvacrol, 584µg mL-1 e thymol, 427µg mL-1. Isolados com diferentes perfis de susceptibil idade aos antimicrobianos usados no tratamento de mastite bovina, quando subagrupados, foram inibidos por concentrações semelhantes de OE . Estes resultados confirmam a atividade antimicrobiana de OE e algumas frações majoritárias.

  6. Biodegradation of glyphosate herbicide by Salinicoccus spp isolated from Qom Hoze-soltan lake, Iran

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    Yaser Sharifi

    2015-01-01

    Full Text Available Background: Glyphosate (N-phosphonomethyl Glycine is an organophosphorus pesticide with dangerous effects on the environment. In this study, the biodegradation of glyphosate herbicide by halophilic bacteria isolated from Qom Hoze-Soltan Lake has been investigated. Methods: After sampling and bacterial isolation, native halophilic strains grown in the presence of glyphosate at a wavelength of 660 nm and also the disappearance of the glyphosate in the plates at a wavelength of 220 nm were determined and the dominant bacteria were isolated. Biochemical, molecular (according to the 16S rRNA sequence, antibiotic, and the Minimum Inhibitory Concentration (MIC test was performed for the dominant bacteria. Analysis of the remaining glyphosate herbicide was performed by HPLC analysis after derivation with FMOC-Cl. Results: According to the results of the biochemical, antibiotic and molecular 16S rRNA tests, the native halophilic isolates with the ability to biodegrade glyphosate were gram positive cocci very similar to Salinicoccusspp. The results of HPLC showed that Salinicoccusspp is able to biodegrade glyphosate herbicide. Conclusion: The native bacteria in Qom Hoze-soltanlake, Iran can be used for biodegradation of glyphosate herbicide.

  7. Enterotoxigenic Genes in strains of Staphylococcus spp., isolated from cheese made in Pamplona-Colombia

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    Fanny Herrera A.

    2015-01-01

    Full Text Available Objective. To determine the incidence of coagulase-positive strains of enterotoxigenic Staphylococcus in doble crema (double cream cheese samples produced in Pamplona. Materials and methods. Bacterial isolation was performed following the routine method for coagulase positive Staphylococcus provided by the Colombian Technical Standard 4779, by using Baird Parker medium with confirmation of typical colonies by performing the coagulase test. Detection of genes for principal enterotoxins was done by PCR. Results. The prevalence of coagulase positive Staphylococcus in cheese samples was 31%, with 27% of the samples failing to meet the requirements of the NTC 750. In 24.6% of the studied isolates, genes for enterotoxin production were detected. The presence, in the isolated strains, of genes for SEB, SEA and SED was 18.5%, 4.6% and 3.0%, respectively. Conclusions. The significant presence of enterotoxigenic genes found in the isolates obtained from samples of double cream cheese made in Pamplona, suggests an important hazard to the health of consumers.

  8. Characterization of clinical and environmental Mycobacterium avium spp. isolates and their interaction with human macrophages

    Science.gov (United States)

    Members of the Mycobacterium avium complex (MAC) are naturally occurring bacteria in the environment. A link has been suggested between M. avium strains in drinking water and clinical isolates from infected individuals. There is a need to develop new screening methodologies tha...

  9. THE INFLUENCE OF NATURAL ESSENTIAL OILS ON THE GROWTH OF PHYTOPHTHORA SPP. ISOLATED FROM PELARGONIUM CUTTINGS

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    Marcelina Machura

    2017-10-01

    Full Text Available Ornamental plants play an important role in human life. Plants positively influence the psyche and improve the well-being of people around them. They produce oxygen, provide a barrier to dust and noise, lower the temperature and increase air humidity, thereby positively impacting the microclimate. The unmatched appeal of pelargonium, ease of cultivation and care, abundance of flowering from spring to late autumn and its decorative qualities make it a universal application. The aim of the study was to isolate the microorganisms that inhabit the cuttings of pelargonium, identify fungal isolates, investigate the pathogenicity of selected isolates and evaluate the influence of certain essential oils (Carum carvi L. essential oils, Citrus limon L. essential oils, Citrus reticulatae aetheroleum essential oils, essential oil of tea tree in in vitro circumstances on the linear growth of the mycelium: Phytophthora cryptogea, Phytophthora nicotianae var. nicotianae. Previcur Energy 840 SL was used as a standard chemical protection. The most numerous isolated fungi were: Phytophthora, Botrytis, Cylindrocladium, Alternaria and Cylindrocarpon. The highest efficiency in relation to Phytophthora cryptogea characterized the Citrus limon L. essential oils (concentration 0.1% and 1% and Carum carvi L. essential oil (concentration 1%.

  10. [Mycotic aneurism in aortic arch by Aspergillus fumigatus: contribution of a case and review of the literature].

    Science.gov (United States)

    Burón Fernández, M R; Oruezábal Moreno, M J

    2005-09-01

    The micotic aneurisms by Aspergillus are rare and usually appear in the context of an invasive pulmonary aspergilosis, or by septicum embolism or direct extension from the lungs, for that reason the location the more frequents is in aortic arch and the ascending aorta.8 cases of micotic aneurisms by Aspergillus spp. have been described in literature between 1966 and 2000, being the most frequent location the ascending aorta or the aortic arch. The Aspergillus fumigatus is the isolated species with more frequency, affecting mainly to patients undergoing inmunosupression. The diagnosis of a micotic aneurism requires a high clinical suspicion, given to its peculiarity and the presence of inespecific symptoms, being frequently an accidental finding in an invasive pulmonary aspergilosis.The case of a patient with a micotic aneurism by A. fumigatus appears and we reviewed the similar cases previously disclosed.

  11. Aflatoxin B1-producing Aspergillus in sun-dried medicinal plant materials

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    Chinaputi, A.

    2001-10-01

    Full Text Available Fifty sun-dried medicinal plants were obtained from fraditional drug stores in Songkhla Province, Thailand, and examined for Aspergillus and aflatoxin B1. 288 isolates of Aspergillus were obtaines by standard blotter plate and 25 species were identified. The most common species were A. niger with 99 isolates, A. Flavus 84 isolates, A. terreus 33 isolates, A. oryzae 25 isolates, A.nidulans (Emericella nidulans 10 isolates, A fumigatus 9 isolates and A. chevalieri (Eurotium chevalieri 8 isolates. The other species[A. alliaceus, A.auricomus, A. carbonarius, A. carneus, A. clavatus, A. fisheri(Sartorya fumigata, A. janus, A. melleus,A. ochraceus, A. phoencis, A. sparsus, A. terricola, A. thomii, A. versicolor, A. wentii and Aspergillus sp.1-3] each had 1-2 siolates. Ofthe 50 different plants examined,9 had no trace of Aspergillus, namely Cinnamomum zeylanicum, Illicium verum, Andrographis paniculate, Carthamus tinctorius, Eugenia caryophyllus, Elettaria cardomomum, Coriandrum sativum, Curcuma longa and Cassia garrettiana. The highest number of species(9 of Aspergillus was found on Rauvolfia serpentina.The ability of Aspergillus to form aflatoxin was determined in coconut milk agar by observing the intensity of blue fluorescence in agar surrounding the colonies under ultraviolet light and the yellow pigment under the colonies. The results showed the production of aflatoxin was limited to the one species, A. flavus, from which 84 isolates produced aflatoxin in 57 isolates(67.8%.Aflatoxin B1. production was confirmed by culturing fluorescencing isolates of A. flavus in coconut nilk broth and detecting by ELISA technique. Aflatoxin B1. showed increasing production after 2 days, stabilizing at 3-4 days, and the decreasing after 5-6 days. Aflatoxin B1. could not be detected from nonfluorescencing isolates.The morphological characteristics of the aflatoxin B1. -producing and non-producing strains of A. flavus were similar under light microscope and

  12. Morphological and molecular identification of four Brazilian commercial isolates of Pleurotus spp. and cultivation on corncob

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    Nelson Menolli Junior

    2010-04-01

    Full Text Available The species of Pleurotus have great commercial importance and adaptability for growth and fructification within a wide variety of agro-industrial lignocellulosic wastes. In this study, two substrates prepared from ground corncobs supplemented with rice bran and charcoal were tested for mycelium growth kinetics in test tubes and for the cultivation of four Pleurotus commercial isolates in polypropylene bags. The identification of the isolates was based on the morphology of the basidiomata obtained and on sequencing of the LSU rDNA gene. Three isolates were identified as P. ostreatus, and one was identified as P. djamor. All isolates had better in-depth mycelium development in the charcoal-supplemented substrate. In the cultivation experiment, the isolates reacted differently to the two substrates. One isolate showed particularly high growth on the substrate containing charcoal.Espécies de Pleurotus têm grande importância comercial e adaptabilidade para crescimento e frutificação em uma ampla variedade de resíduos agro-industriais lignocelulósicos. Neste trabalho foram testados dois substratos à base de sabugo de milho triturado, suplementados com farelo de arroz e carvão vegetal, para avaliação da cinética de crescimento micelial em tubos de ensaio e produção em sacos de polipropileno, utilizando quatro isolados comerciais. O estudo taxonômico foi realizado com a análise da morfologia dos basidiomas obtidos em cultivo e pelo seqüenciamento do gene nLSU do DNAr, para certificar a identificação taxonômica. Os isolados tiveram melhor desenvolvimento micelial em profundidade no substrato suplementado com carvão vegetal. Em relação à produção, os isolados reagiram de formas distintas em função dos substratos, sendo significativamente melhor o substrato contendo carvão. Três isolados foram identificados como P. ostreatus e o outro foi identificado como P. djamor.

  13. Resistance to linezolid in Staphylococcus spp. clinical isolates associated with ribosomal binding site modifications: novel mutation in domain V of 23S rRNA.

    Science.gov (United States)

    Musumeci, Rosario; Calaresu, Enrico; Gerosa, Jolanda; Oggioni, Davide; Bramati, Simone; Morelli, Patrizia; Mura, Ida; Piana, Andrea; Are, Bianca Maria; Cocuzza, Clementina Elvezia

    2016-10-01

    Linezolid is the main representative of the oxazolidinones, introduced in 2000 in clinical practice to treat severe Gram-positive infections. This compound inhibits protein synthesis by binding to the peptidyl transferase centre of the 50S bacterial ribosomal subunit. The aim of this study was to characterize 12 clinical strains of linezolid-resistant Staphylococcus spp. isolated in Northern Italy. All isolates of Staphylococcus spp. studied showed a multi-antibiotic resistance phenotype. In particular, all isolates showed the presence of the mecA gene associated with SSCmec types IVa, V or I. Mutations in domain V of 23S rRNA were shown to be the most prevalent mechanism of linezolid resistance: among these a new C2551T mutation was found in S. aureus, whilst the G2576T mutation was shown to be the most prevalent overall. Moreover, three S. epidermidis isolates were shown to have linezolid resistance associated only with alterations in both L3 and L4 ribosomal proteins. No strain was shown to harbor the previously described cfr gene. These results have shown how the clinical use of linezolid in Northern Italy has resulted in the selection of multiple antibiotic-resistant clinical isolates of Staphylococcus spp., with linezolid resistance in these strains being associated with mutations in 23S rRNA or ribosomal proteins L3 and L4.

  14. Effects of temperature, water activity and incubation time on fungal growth and aflatoxin B1 production by toxinogenic Aspergillus flavus isolates on sorghum seeds.

    Science.gov (United States)

    Lahouar, Amani; Marin, Sonia; Crespo-Sempere, Ana; Saïd, Salem; Sanchis, Vicente

    2016-01-01

    Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37°C), water activity (aw, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 a(w) at 37°C for two of the isolates. The minimum aw needed for mycelial growth was 0.91 at 25 and 37°C. At 15°C, only isolate 8 grew at 0.99 a(w). Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 a(w)). Aflatoxin production was not observed at 15°C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  15. Reclassification of Borrelia spp. isolated in South Korea using Multilocus Sequence Typing.

    Science.gov (United States)

    Park, Kyung-Hee; Choi, Yeon-Joo; Kim, Jeoungyeon; Park, Hye-Jin; Song, Dayoung; Jang, Won-Jong

    2018-05-31

    Using Borrelia isolated from South Korea, we evaluated by MLST and three intergenic genes (16S rRNA, ospA, and 5S-23S IGS) typing to analyze the relationship between host and vector and molecular background. Using the MLST analysis, we identified B. afzelii, B. yangtzensis, B. garinii, and B. bavariensis. This study was first report of the identification of B. yangtzensis using the MLST in South Korea.

  16. Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico

    Science.gov (United States)

    Zavala-Norzagaray, Alan A.; Aguirre, A. Alonso; Velazquez-Roman, Jorge; Flores-Villaseñor, Héctor; León-Sicairos, Nidia; Ley-Quiñonez, C. P.; Hernández-Díaz, Lucio De Jesús; Canizalez-Roman, Adrian

    2015-01-01

    The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL), Baja California Sur (BCS), Mexico (Pacific Ocean) and the lagoon system of Navachiste (LSN) and Marine Area of Influence (MAI), Guasave, Sinaloa (Gulf of California). A total of 34 black turtles (Chelonia mydas agassizii) were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea) were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%), V. parahaemolyticus in 17/64 (26%), and V. cholerae in 6/64 (9%). However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI). Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4%) belonged to the pathogenic strains (tdh+ gene) and 2/17 (11.7%) had the pandemic clone (tdh+ and toxRS/new+). Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66%) the accessory cholera enterotoxin gene (ace) was identified but without virulence gene zot, ctxA, and ctxB. Of the isolated V. parahaemolyticus, V. cholerae, and V. alginolyticus strains, 94.1, 33.4, and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin), respectively. In conclusion, the presence of several potential (toxigenic) human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico. PMID:26161078

  17. Isolation, Characterization, and Antibiotic Resistance of Vibrio spp. in Sea Turtles from Northwestern Mexico

    Directory of Open Access Journals (Sweden)

    Alan A. eZavala-Norzagaray

    2015-06-01

    Full Text Available The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL, Baja California Sur, Mexico (Pacific Ocean and the lagoon system of Navachiste (LSN and Marine Area of Influence (MAI, Guasave, Sinaloa (Gulf of California. A total of 34 black turtles (Chelonia mydas agassizii were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%, V. parahaemolyticus in 17/64 (26% and V. cholerae in 6/64 (9%,. However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI. Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4% belonged to the pathogenic strains (tdh+ gene and 2/17 (11.7% had the pandemic clone (tdh+ and toxRS/new+. Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66% the accessory cholera enterotoxin gene (ace was identified but without virulence gene zot, ctxA and ctxB. Of the isolated V. parahaemolyticus, V. cholerae and V. alginolyticus strains, 94.1%, 33.4% and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin, respectively. In conclusion, the presence of several potential (toxigenic human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico.

  18. Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico.

    Science.gov (United States)

    Zavala-Norzagaray, Alan A; Aguirre, A Alonso; Velazquez-Roman, Jorge; Flores-Villaseñor, Héctor; León-Sicairos, Nidia; Ley-Quiñonez, C P; Hernández-Díaz, Lucio De Jesús; Canizalez-Roman, Adrian

    2015-01-01

    The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL), Baja California Sur (BCS), Mexico (Pacific Ocean) and the lagoon system of Navachiste (LSN) and Marine Area of Influence (MAI), Guasave, Sinaloa (Gulf of California). A total of 34 black turtles (Chelonia mydas agassizii) were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea) were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%), V. parahaemolyticus in 17/64 (26%), and V. cholerae in 6/64 (9%). However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI). Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4%) belonged to the pathogenic strains (tdh (+) gene) and 2/17 (11.7%) had the pandemic clone (tdh (+) and toxRS/new (+)). Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66%) the accessory cholera enterotoxin gene (ace) was identified but without virulence gene zot, ctxA, and ctxB. Of the isolated V. parahaemolyticus, V. cholerae, and V. alginolyticus strains, 94.1, 33.4, and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin), respectively. In conclusion, the presence of several potential (toxigenic) human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico.

  19. Various Ready-to-Eat Products from Retail Stores Linked to Occurrence of Diverse Listeria monocytogenes and Listeria spp. Isolates.

    Science.gov (United States)

    Vongkamjan, Kitiya; Fuangpaiboon, Janejira; Turner, Matthew P; Vuddhakul, Varaporn

    2016-02-01

    Listeriosis outbreaks have been associated with a variety of foods. This study investigated the prevalence and diversity of Listeria monocytogenes and Listeria spp. in ready-to-eat (RTE) products and evaluated the performance of a rapid detection method, the 3M molecular detection assay for L. monocytogenes (MDA-LM), for detection of L. monocytogenes. Assay results were compared with those obtained using the U.S. Food and Drug Administration standard culture method described in the Bacteriological Analytical Manual. Products (n = 200) were purchased from retail stores: 122 aquatic products, 22 products of animal origin, 18 vegetarian products, 15 deli meat products, 13 salad and vegetable products, 4 desserts, 2 egg-based products, and 4 other products. L. monocytogenes prevalence was comparable with both methods. Overall, 15 (7.5%) of 200 samples were positive for L. monocytogenes: 3% of aquatic products, 1.5% of products of animal origin, 1% of vegetarian products, and 2% of deli meat products. Compared with the standard culture method, the sensitivity, specificity, and the accuracy of the MDA-LM were 86.7% (95% confidence interval, 58.4 to 97.7%), 98.4% (95% confidence interval, 95.0 to 99.6%), and 97.5%, respectively. Using the culture-based method, 18 (9%) of 200 samples were positive for Listeria species other than L. monocytogenes. Listeria isolates from these samples were classified into nine allelic types (ATs). The majority of isolates were classified as ATs 58 and 74, which were identified as L. monocytogenes lineages I and IV, respectively. Listeria innocua and Listeria welshimeri also were represented by isolates of multiple ATs. The MDA-LM is a rapid and reliable technique for detecting L. monocytogenes in various RTE foods. Further study is needed to develop effective control strategies to reduce L. monocytogenes contamination in RTE foods.

  20. Production of enterotoxins of Staphylococcus spp. isolated from samples of sheep milk

    Directory of Open Access Journals (Sweden)

    František Zigo

    2014-02-01

    Full Text Available In our study was followed occurrence of mastitis in herd of 430 sheep of breed zoslachtena valaska with hand milking technology examined two times during one lactation season. Individual examination consisted from clinical examination of udder and microbiological examination of milk samples. By PCR was determined presence of genes coding production of enterotoxins, and by ELISA methods production individual types of enterotoxins. From individual forms of mastitis were frequently detected subacute (6.7%, subclinical (5.7% and acute (2.9%. The coagulase-negative staphylococci (CNS were identified in 102 (65.4% from all 156 positive isolates. The CNS and S. aureus caused subacute (5.1%, subclinical (3.9% and acute (2.4% forms of mastitis. The most frequently isolated were S. epidermidis, followed by S. chromogenes and S. xylosus from ewes with subacute and subclinical mastitis. From acute and chronical forms of mastitis were  predominantly isolated S. aureus, S. uberis and S. epidermidis. The production of staphylococcal enterotoxins (SE - SEA, SEB, SEC, SED and the presence of genes sec (3, sea (2, seb (2 and sed (2 were determined in S. aureus, S. epidermidis, S. schleiferi and S. chromogenes, respectively. The results suggested on the high occurrence (12.4% of subacute and subclinical forms. Confirmed production of enterotoxins and presence of genes coding their production present a risk for human health and decreased a quality of milk and products from sheep´s milk.

  1. Commensal Staphylococcus spp., Acinetobacter spp. and ...

    African Journals Online (AJOL)

    ANTHONY

    2012-07-31

    Jul 31, 2012 ... Intermittent assessment of resistance genes in the ecosystem should be ..... among resistant Acinetobacter spp. isolated from integrated fish .... independent studies on the emerging phylogenetic view of bacterial .... Functional.

  2. Molecular characterization of Aspergillus fumigatus isolated from raw cow milk in Argentina: Molecular typing of A. fumigatus from raw cow milk.

    Science.gov (United States)

    Pena, Gabriela A; Alonso, Verónica; Manini, Marina Velasco; Pellegrino, M; Cavaglieri, Lilia R

    2018-06-20

    Aspergillus fumigatus, the major etiological agent of human and animal aspergillosis, is a gliotoxinogenic species into section Fumigati commonly found in contaminated animal environments. In dairy herds, exposed areas of lactating cows, as mammalian glandule, can be easily contaminated by them. This study was aimed to identify A. fumigatus sensu lato strains (identified based on morphology) isolated from raw cow milk at species level, by morphological and molecular techniques, and to estimate their genetic variability. Forty-five A. fumigatus strains showed similar RAPD profiles (generated with PELF and URP1F primers) to each other and to A. fumigatus sensu stricto reference strains; also, they were almost identical to clinical human and feed-borne A. fumigatus strains included in the assay, since their similarity coefficient ranged from 0.7 to 1.00. Therefore, all strains were characterized as belonging to A. fumigatus sensu stricto species. This result was supported by sequencing the benA gene of selected strains and by maximum parsimony analysis. In addition, RAPD fingerprinting demonstrated intra-specific genetic variability into the A. fumigatus sensu stricto cluster. The results found in this study strengthen the fact that A. fumigatus sensu stricto is the predominant species in the Aspergillus section Fumigati found in animal environments such as dairy herd environments, while other species such as A. novofumigatus, A. fumigatiaffinis, A. udagawae and A. lentulus may be rarely isolated. Since no differences between animal and human strains were observed they can become pathogenic also for farm handlers'. Moreover, the presence of A. fumigatus sensu stricto in raw cow milk is probably a very important risk factor since milk and its by-products are generally indented for human consumption, then gliotoxin could be transferred to them. Copyright © 2018. Published by Elsevier B.V.

  3. Production of 3,4-dihydroxy L-phenylalanine by a newly isolated Aspergillus niger and parameter significance analysis by Plackett-Burman design

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    Haq I

    2010-12-01

    Full Text Available Abstract Background The amino acid derivative 3,4-dihydroxy L-phenylalanine (L-dopa is gaining interest as a drug of choice for Parkinson's disease. Aspergillus oryzae is commonly used for L-dopa production; however, a slower growth rate and relatively lower tyrosinase activity of mycelia have led to an increasing interest in exploiting alternative fungal cultures. In the present investigation, we report on the microbiological transformation of L-tyrosine to L-dopa accomplished by a newly isolated filamentous fungus Aspergillus niger. Results The culture A. niger (isolate GCBT-8 was propagated in 500 ml Erlenmeyer flasks and the pre-grown mycelia (48 h old were used in the reaction mixture as a source of enzyme tyrosinase. Grinded mycelia gave 1.26 fold higher L-dopa production compared to the intact at 6% glucose (pH 5.5. The rate of L-tyrosine consumption was improved from 0.198 to 0.281 mg/ml. Among the various nitrogen sources, 1.5% peptone, 1% yeast extract and 0.2% ammonium chloride were optimized. The maximal L-dopa was produced (0.365 mg/ml at 0.3% potassium dihydrogen phosphate with L-tyrosine consumption of 0.403 mg/ml. Conclusion Over ~73% yield was achieved (degree of freedom 3 when the process parameters were identified using 2k-Plackett-Burman experimental design. The results are highly significant (p ≤ 0.05 and mark the commercial utility (LSD 0.016 of the mould culture which is perhaps the first ever report on L-dopa production from A. niger.

  4. Molecular characterization and classification of Trypanosoma spp. Venezuelan isolates based on microsatellite markers and kinetoplast maxicircle genes.

    Science.gov (United States)

    Sánchez, E; Perrone, T; Recchimuzzi, G; Cardozo, I; Biteau, N; Aso, P M; Mijares, A; Baltz, T; Berthier, D; Balzano-Nogueira, L; Gonzatti, M I

    2015-10-15

    Livestock trypanosomoses, caused by three species of the Trypanozoon subgenus, Trypanosoma brucei brucei, T. evansi and T. equiperdum is widely distributed throughout the world and constitutes an important limitation for the production of animal protein. T. evansi and T. equiperdum are morphologically indistinguishable parasites that evolved from a common ancestor but acquired important biological differences, including host range, mode of transmission, distribution, clinical symptoms and pathogenicity. At a molecular level, T. evansi is characterized by the complete loss of the maxicircles of the kinetoplastic DNA, while T. equiperdum has retained maxicircle fragments similar to those present in T. brucei. T. evansi causes the disease known as Surra, Derrengadera or "mal de cadeiras", while T. equiperdum is the etiological agent of dourine or "mal du coit", characterized by venereal transmission and white patches in the genitalia. Nine Venezuelan Trypanosoma spp. isolates, from horse, donkey or capybara were genotyped and classified using microsatellite analyses and maxicircle genes. The variables from the microsatellite data and the Procyclin PE repeats matrices were combined using the Hill-Smith method and compared to a group of T. evansi, T. equiperdum and T. brucei reference strains from South America, Asia and Africa using Coinertia analysis. Four maxicircle genes (cytb, cox1, a6 and nd8) were amplified by PCRfrom TeAp-N/D1 and TeGu-N/D1, the two Venezuelan isolates that grouped with the T. equiperdum STIB841/OVI strain. These maxicircle sequences were analyzed by nucleotide BLAST and aligned toorthologous genes from the Trypanozoon subgenus by MUSCLE tools. Phylogenetic trees were constructed using Maximum Parsimony (MP) and Maximum Likelihood (ML) with the MEGA5.1® software. We characterized microsatellite markers and Procyclin PE repeats of nine Venezuelan Trypanosoma spp. isolates with various degrees of virulence in a mouse model, and compared them to a

  5. Species distribution and antifungal susceptibility of Candida spp. isolated from superficial candidiasis in outpatients in Iran.

    Science.gov (United States)

    Razzaghi-Abyaneh, M; Sadeghi, G; Zeinali, E; Alirezaee, M; Shams-Ghahfarokhi, M; Amani, A; Mirahmadi, R; Tolouei, R

    2014-06-01

    Candidiasis is the most prevalent fungal infection affecting human and animals all over the world. This study represents the epidemiological aspects of superficial candidiasis in outpatients and in vitro antifungal susceptibility of etiologic Candida species. Clinical samples were taken from 173 patients including skin and nail scrapings (107; 61.8%), vaginal discharge (28; 16.2%), sputum (20; 11.6%), oral swabs (7; 4.0%), bronchoalveolar lavage (6; 3.5%) and 1 specimen (0.6%) of each eye tumor, gastric juice, urine, biopsy and urinary catheter and confirmed as candidiasis by direct microscopy, culture and histopathology. Susceptibility patterns of the isolated Candida species were determined using the disk diffusion and broth microdilution methods. Among 173 Candida isolates, C. albicans (72.3%) was the most prevalent species followed by C. parapsilosis (11.5%). Other identified species were C. glabrata, C. krusei, C. tropicalis, C. guilliermondii, C. intermedia and C. sake. Majority of the Candida isolates were susceptible to fluconazole (95.4%) followed by 5-flucytosine (89.6%), voriconazole (78.6%) itraconazole (48.0%) and ketoconazole (42.8%). Caspofungin was the most potent antifungal drug against C. albicans (MICs; 0.062-1 μg/mL), ketoconazole for C. parapsilosis and C. tropicalis (MICs; 0.031-0.25 μg/mL) and itraconazole for C. krusei, C. glabrata and C. guilliermondii (MICs; 0.031-1 μg/mL). This study reinforces the significance of superficial candidiasis as an important fungal infection with multiple clinical presentations. Our results further indicate that susceptibility testing to commonly used antifungals is crucial in order to select the appropriate therapeutic strategies which minimize complications while improving patients' life. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  6. Glucanase and Chitinase from Some Isolates of Endophytic Fungus Trichoderma spp.

    Science.gov (United States)

    Prasetyawan, Sasangka; Sulistyowati, Lilik; Aulanni'am

    2018-01-01

    Endophytic fungi are those fungi that are able to grow in plant tissue without causing symptoms of disease. It is thought that these fungi may confer on the host plants degree of resistance to parasitic invasion. Endophytic fungi have been isolated from stem tissue and these fungi are known to be antagonistic to pathogenic fungi. These endophytes produce chitinase and β-1,3-glucanase enzymes. Based on the fact that chitin and β-1,3-glucan are the main skeletal polysaccharides of the cell walls of fungal patogen. The aim of this research is to do potential test on some of isolates of Trichoderma’s endophytic (L-1, L-2, Is-1, Is-2 and Is-7) in the chitinase and β-1,3-glucanase activity in effort to determine endophytic which be chossen to be gene resource for the next research. The gene will be transformed to citrus plant japanese citroen in effort to make citrus plant transgenic resistance to phytopatogenic invasion. The result of this research is endofit namely L-1 is the most potential endophytic fungi with chitinase activities is 4,8 10-2 Unit and glucanase 24,2. 1012 Unit. The addition of chitin and cell wall of phytophtora causes chitinase activity significantly increase, and also addition of laminarin and cell wall of phytophtora makes glucanase activity increase.

  7. Characterization of Brugmansia mosaic virus Isolated from Brugmansia spp. in Korea

    Directory of Open Access Journals (Sweden)

    Chung Youl Park

    2014-12-01

    Full Text Available In May 2013, an angel’s trumpet leaves showing mosaic and malformation symptoms were collected from Suwon city, Gyeonggi-do. An analysis of the collected sample by transmission electron microscopy observation showed filamentous rod particles of 720-800 nm in length. On the basis of the these observations, we performed PCR against three reported Potyviruses (Brugmansia mosaic virus, Colombian datura virus and Brugmansia suaveolens mottle virus, and the sample was positive for BruMV. Pathogenicity and host range test of BruMV was determined by mechanical inoculation. Solanaceae (tobacco, tomato and eggplant and Amaranthaceae (ground cherry appeared typical virus symptoms. To determine coat protein of this virus, we designed specific primer pairs, and performed PCR amplification, cloning, and sequencing. Phylogenetic analysis showed that BruMV-SW was most closely related to BruMV isolate SK. Comparison of the BruMV-SW coat protein nucleotide sequences showed 92% to 99% identities to the other BruMV isolates.

  8. A rapid and inexpensive method for isolation of total DNA from Trichoderma spp (Hypocreaceae).

    Science.gov (United States)

    Vazquez-Angulo, J C; Mendez-Trujillo, V; González-Mendoza, D; Morales-Trejo, A; Grimaldo-Juarez, O; Cervantes-Díaz, L

    2012-05-15

    Extraction of high-quality genomic DNA for PCR amplification from filamentous fungi is difficult because of the complex cell wall and the high concentrations of polysaccharides and other secondary metabolites that bind to or co-precipitate with nucleic acids. We developed a modified sodium dodecyl sulfate/phenol protocol, without maceration in liquid nitrogen and without a final ethanol precipitation step. The A(260/280) absorbance ratios of isolated DNA were approximately 1.7-1.9, demonstrating that the DNA fraction is pure and can be used for analysis. Additionally, the A(260/230) values were higher than 1.6, demonstrating negligible contamination by polysaccharides. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. The main advantages of the method are that the mycelium is directly recovered from culture medium and it does not require the use of expensive and specialized equipment.

  9. Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp. isolated from dairy and pharmaceutical products.

    Science.gov (United States)

    D'Aimmo, Maria Rosaria; Modesto, Monica; Biavati, Bruno

    2007-04-01

    The outlines of antibiotic resistance of some probiotic microorganisms were studied. This study was conducted with the double purpose of verifying their ability to survive if they are taken simultaneously with an antibiotic therapy and to increase the selective properties of suitable media for the isolation of samples containing mixed bacterial populations. We isolated from commercial dairy and pharmaceutical products, 34 strains declared as probiotics, belonging to the genera Bifidobacterium and Lactobacillus, and 21 strains of starter culture bacteria. All the microorganisms have been compared by electrophoresis of the soluble proteins for the purpose of identifying them. A Multiplex-PCR with genus- and species-specific primers was used to detect for Bifidobacterium animalis subsp. lactis presence. All bifidobacteria were B. animalis subsp. lactis except one Bifidobacterium longum. Sometimes the identification showed that the used strain was not the one indicated on the label. The lactobacilli were Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus delbrueckii subsp. bulgaricus. The streptococci were all Streptococcus thermophilus. The minimal inhibitory concentration (MIC) of 24 common antibiotic substances has been valued by the broth microdilution method. All tested strains were susceptible to ampicillin, bacitracin, clindamycin, dicloxacillin, erytromycin, novobiocin, penicillin G, rifampicin (MIC(90) ranging from 0.01 to 4 microg/ml); resistant to aztreonam, cycloserin, kanamycin, nalidixic acid, polymyxin B and spectinomycin (MIC(90) ranging from 64 to >1000 microg/ml). The susceptibility to cephalothin, chloramphenicol, gentamicin, lincomycin, metronidazole, neomycin, paromomycin, streptomycin, tetracycline and vancomycin was variable and depending on the species.

  10. Prevalence of Thermotolerant Campylobacter spp. in Chicken Meat in Croatia and Multilocus Sequence Typing of a Small Subset of Campylobacter jejuni and Campylobacter coli Isolates

    Directory of Open Access Journals (Sweden)

    Andrea Humski

    2016-01-01

    Full Text Available In order to detect thermotolerant Campylobacter spp., 241 samples of fresh chicken meat, at retail in Croatia, were analysed according to a standard method, followed by biochemical test and molecular polymerase chain reaction/restriction enzyme analysis for exact species determination. Campylobacter spp. prevalence was 73.86 %. Campylobacter jejuni and Campylobacter coli were isolated from 53.53 and 15.35 % of the samples, respectively. In 4.98 % of isolates thermotolerant Campylobacter spp. were not determined. The multi locus sequence typing method was used to evaluate genetic diversity of eight Campylobacter jejuni and four Campylobacter coli isolates. To our knowledge, these results of genotyping provided the first data on the presence of sequence types (STs and clonal complexes (CCs of Campylobacter jejuni and C. coli isolates in Croatia. By applying the multilocus sequence typing, a new allele of tkt gene locus was discovered and marked tkt508. The C. jejuni ST 6182 and C. coli ST 6183 genotypes were described for the fi rst time, and all other identified genotypes were clustered in the previously described sequence types and clonal complexes. These findings provide useful information on the prevalence and epidemiology of Campylobacter jejuni and C. coli in Croatia.

  11. EFFECT OF HIGHLY ACTIVE ANTIRETROVIRAL THERAPY ON VAGINAL Candida spp. ISOLATION IN HIV-INFECTED COMPARED TO HIV-UNINFECTED WOMEN

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    Silvia de Souza Dantas ALCZUK

    2015-04-01

    Full Text Available Vulvovaginal candidiasis (VVC in HIV-infected women contributed to the impairment of their quality of life. The aim of this study was to evaluate the effect of highly active antiretroviral therapy (HAART use on the vaginal Candida spp. isolation in HIV-infected compared to HIV-uninfected women. This cross-sectional study included 178 HIV-infected (HIV group and 200 HIV-uninfected women (control that were studied at the Specialized Assistance Service (SAE for sexually transmitted diseases (STD/AIDS of the city of Maringá, Brazil, from April 1 to October 30, 2011. The yeasts were isolated and identified by phenotypic and molecular methods. The in vitro antifungal susceptibility to fluconazole, itraconazole, nystatin and amphotericin B was tested by the reference microdilution method. Higher frequencies of total vaginal Candida spp. isolation were found in the HIV-infected group than in the control group. However, both groups showed a similar frequency of colonization and VVC. Although C. albicans was the most frequent and sensitive to azolics and polyenes in both HIV-infected and uninfected women, the emerging resistance of C. glabrata to amphotericin B in the HIV-infected women was observed. Although higher frequency of vaginal Candida spp. isolation had been observed in the HIV-infected than in HIV-uninfected women, colonization and VVC showed similar frequency in both groups, indicating that HAART appears to protect against vaginal colonization and VVC.

  12. Isolation of Cronobacter spp. (formerly Enterobacter sakazakii) from infant food, herbs and environmental samples and the subsequent identification and confirmation of the isolates using biochemical, chromogenic assays, PCR and 16S rRNA sequencing.

    Science.gov (United States)

    Jaradat, Ziad W; Ababneh, Qotaiba O; Saadoun, Ismail M; Samara, Nawal A; Rashdan, Abrar M

    2009-10-27

    Cronobacter spp. (formerly Enterobacter sakazakii), are a group of Gram-negative pathogens that have been implicated as causative agents of meningitis and necrotizing enterocolitis in infants. The pathogens are linked to infant formula; however, they have also been isolated from a wide range of foods and environmental samples. In this study, 233 samples of food, infant formula and environment were screened for the presence of Cronobacter spp. in an attempt to find its source. Twenty nine strains were isolated from samples of spices, herbs, infant foods, and dust obtained from household vacuum cleaners. Among the 76 samples of infant food, infant formula, milk powder and non-milk dairy products tested, only one sample of infant food contained Cronobacter spp. (1.4%). The other Cronobacter spp. isolates recovered include two from household vacuum dust, and 26 from 67 samples of herbs and spices. Among the food categories analyzed, herbs and spices harbored the highest number of isolates, indicating plants as a possible reservoir of this pathogen. Initial screening with API 20E test strips yielded 42 presumptive isolates. Further characterization using 3 chromogenic media (alpha-MUG, DFI and EsPM) and 8 sets of PCR primers detecting ITS (internal transcribed spacer sequences), 16S rRNA, zpx, gluA, gluB, OmpA genes followed by nucleotide sequencing of some PCR amplicons did not confirm the identity of all the isolates as none of the methods proved to be free of both false positives or false negatives. The final confirmation step was done by 16S rRNA sequence analysis identifying only 29 of the 42 isolates as Cronobacter spp. Our studies showed that Cronobacter spp. are highly diverse and share many phenotypic traits with other Enterobacteriaceae members highlighting the need to use several methods to confirm the identity of this pathogen. None of the biochemical, chromogenic or PCR primers proved to be a reliable method for confirmation of the identity of the isolates

  13. Isolation of Cronobacter spp. (formerly Enterobacter sakazakii from infant food, herbs and environmental samples and the subsequent identification and confirmation of the isolates using biochemical, chromogenic assays, PCR and 16S rRNA sequencing

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    Samara Nawal A

    2009-10-01

    Full Text Available Abstract Background Cronobacter spp. (formerly Enterobacter sakazakii, are a group of Gram-negative pathogens that have been implicated as causative agents of meningitis and necrotizing enterocolitis in infants. The pathogens are linked to infant formula; however, they have also been isolated from a wide range of foods and environmental samples. Results In this study, 233 samples of food, infant formula and environment were screened for the presence of Cronobacter spp. in an attempt to find its source. Twenty nine strains were isolated from samples of spices, herbs, infant foods, and dust obtained from household vacuum cleaners. Among the 76 samples of infant food, infant formula, milk powder and non-milk dairy products tested, only one sample of infant food contained Cronobacter spp. (1.4%. The other Cronobacter spp. isolates recovered include two from household vacuum dust, and 26 from 67 samples of herbs and spices. Among the food categories analyzed, herbs and spices harbored the highest number of isolates, indicating plants as a possible reservoir of this pathogen. Initial screening with API 20E test strips yielded 42 presumptive isolates. Further characterization using 3 chromogenic media (α-MUG, DFI and EsPM and 8 sets of PCR primers detecting ITS (internal transcribed spacer sequences, 16S rRNA, zpx, gluA, gluB, OmpA genes followed by nucleotide sequencing of some PCR amplicons did not confirm the identity of all the isolates as none of the methods proved to be free of both false positives or false negatives. The final confirmation step was done by 16S rRNA sequence analysis identifying only 29 of the 42 isolates as Cronobacter spp. Conclusion Our studies showed that Cronobacter spp. are highly diverse and share many phenotypic traits with other Enterobacteriaceae members highlighting the need to use several methods to confirm the identity of this pathogen. None of the biochemical, chromogenic or PCR primers proved to be a reliable

  14. Antifungal Activity of Thapsia villosa Essential Oil against Candida, Cryptococcus, Malassezia, Aspergillus and Dermatophyte Species

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    Eugénia Pinto

    2017-09-01

    Full Text Available The composition of the essential oil (EO of Thapsia villosa (Apiaceae, isolated by hydrodistillation from the plant’s aerial parts, was analysed by GC and GC-MS. Antifungal activity of the EO and its main components, limonene (57.5% and methyleugenol (35.9%, were evaluated against clinically relevant yeasts (Candida spp., Cryptococcus neoformans and Malassezia furfur and moulds (Aspergillus spp. and dermatophytes. Minimum inhibitory concentrations (MICs were measured according to the broth macrodilution protocols by Clinical and Laboratory Standards Institute (CLSI. The EO, limonene and methyleugenol displayed low MIC and MFC (minimum fungicidal concentration values against Candida spp., Cryptococcus neoformans, dermatophytes, and Aspergillus spp. Regarding Candida species, an inhibition of yeast–mycelium transition was demonstrated at sub-inhibitory concentrations of the EO (MIC/128; 0.01 μL/mL and their major compounds in Candida albicans. Fluconazole does not show this activity, and the combination with low concentrations of EO could associate a supplementary target for the antifungal activity. The association of fluconazole with T. villosa oil does not show antagonism, but the combination limonene/fluconazole displays synergism. The fungistatic and fungicidal activities revealed by T. villosa EO and its main compounds, associated with their low haemolytic activity, confirm their potential antimicrobial interest against fungal species often associated with human mycoses.

  15. Genetic diversity of Dirofilaria spp. isolated from subcutaneous and ocular lesions of human patients in Ukraine.

    Science.gov (United States)

    Rossi, Alice; Peix, Álvaro; Pavlikovskaya, Tamara; Sagach, Olga; Nikolaenko, Svetlana; Chizh, Nina; Kartashev, Vladimir; Simón, Fernando; Siles-Lucas, Mar

    2015-02-01

    This short communication describes the phylogenetic analysis of 48 Dirofilaria worms isolated from human patients in Ukraine. 102 cases were both of subcutaneous (47; 46.1%) and ocular (54; 52.9%) locations. Worms from 44 patients (15 subcutaneous and 29 ocular) were subjected to DNA extraction and amplification of a specific fragment of the 12S rRNA subunit, and sequences were used for phylogenetic analysis. Results showed that 13.8% of the ocular cases analyzed at molecular level were caused by Dirofilaria immitis. Very few cases of ocular human dirofilariosis due to D. immitis have been described in the literature to date, majority of them attributed to Dirofilaria repens. Our results show that ocular dirofilariosis cannot be excluded in areas of low endemicity for D. repens were D. immitis is also present. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Previously unknown species of Aspergillus.

    Science.gov (United States)

    Gautier, M; Normand, A-C; Ranque, S

    2016-08-01

    The use of multi-locus DNA sequence analysis has led to the description of previously unknown 'cryptic' Aspergillus species, whereas classical morphology-based identification of Aspergillus remains limited to the section or species-complex level. The current literature highlights two main features concerning these 'cryptic' Aspergillus species. First, the prevalence of such species in clinical samples is relatively high compared with emergent filamentous fungal taxa such as Mucorales, Scedosporium or Fusarium. Second, it is clearly important to identify these species in the clinical laboratory because of the high frequency of antifungal drug-resistant isolates of such Aspergillus species. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been shown to enable the identification of filamentous fungi with an accuracy similar to that of DNA sequence-based methods. As MALDI-TOF MS is well suited to the routine clinical laboratory workflow, it facilitates the identification of these 'cryptic' Aspergillus species at the routine mycology bench. The rapid establishment of enhanced filamentous fungi identification facilities will lead to a better understanding of the epidemiology and clinical importance of these emerging Aspergillus species. Based on routine MALDI-TOF MS-based identification results, we provide original insights into the key interpretation issues of a positive Aspergillus culture from a clinical sample. Which ubiquitous species that are frequently isolated from air samples are rarely involved in human invasive disease? Can both the species and the type of biological sample indicate Aspergillus carriage, colonization or infection in a patient? Highly accurate routine filamentous fungi identification is central to enhance the understanding of these previously unknown Aspergillus species, with a vital impact on further improved patient care. Copyright © 2016 European Society of Clinical Microbiology and

  17. Bacillus spp. Isolated from Puba as a Source of Biosurfactants and Antimicrobial Lipopeptides

    Science.gov (United States)

    Perez, Karla J.; Viana, Jaime dos Santos; Lopes, Fernanda C.; Pereira, Jamile Q.; dos Santos, Daniel M.; Oliveira, Jamil S.; Velho, Renata V.; Crispim, Silvia M.; Nicoli, Jacques R.; Brandelli, Adriano; Nardi, Regina M. D.

    2017-01-01

    Several products of industrial interest are produced by Bacillus, including enzymes, antibiotics, amino acids, insecticides, biosurfactants and bacteriocins. This study aimed to investigate the potential of two bacterial isolates (P5 and C3) from puba, a regional fermentation product from cassava, to produce multiple substances with antimicrobial and surface active properties. Phylogenetic analyses showed close relation of isolates P5 and C3 with Bacillus amyloliquefaciens and Bacillus thuringiensis, respectively. Notably, Bacillus sp. P5 showed antimicrobial activity against pathogens such as Listeria monocytogenes and Bacillus cereus, in addition to antifungal activity. The presence of genes encoding pre-subtilosin (sboA), malonyl CoA transacylase (ituD), and the putative transcriptional terminator of surfactin (sfp) were detected in Bacillus sp. P5, suggesting the production of the bacteriocin subtilosin A and the lipopeptides iturin A and surfactin by this strain. For Bacillus sp. C3 the presence of sboA and spas (subtilin) genes was observed by the first time in members of B. cereus cluster. Bacillus sp. P5 showed emulsifying capability on mineral oil, soybean biodiesel and toluene, while Bacillus sp. C3 showed emulsifying capability only on mineral oil. The reduction of the surface tension in culture medium was also observed for strain P5, confirming the production of surface-active compounds by this bacterium. Monoprotonated molecular species and adducts of sodium and potassium ions of surfactin, iturin, and fengycin were detected in the P5 culture medium. Comparative MS/MS spectra of the peak m/z 1030 (C14 surfactin A or C15 surfactin B [M+Na]+) and peak m/z 1079 (C15 iturin [M+Na]+) showed the same fragmentation profile of standards, confirming the molecular identification. In conclusion, Bacillus sp. P5 showed the best potential for the production of antifungal, antibacterial, and biosurfactant substances. PMID:28197131

  18. Molecular identification and characterization of Colletotrichum spp isolates from tahiti lime, tamarillo, and mango

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    Adriana Sanabria

    2010-12-01

    Full Text Available Anthracnose is a very limiting disease affecting production, as well as postharvest quality of numerous fruit crops in Colombia. The current management practices for this disease are partially effective due to limited information about the etiology, the inoculum sources, population structure and variation of the pathogen. A total of 293 Colletotrichum isolates were obtained from symptomatic tissues collected from Tahiti lime, tamarillo and mango orchards. To determine the Colletotrichum species causing the symptoms, amplification, and PCR product analysis for intergenic regions of the ribosomal DNA were conducted. Genetic diversity of the fungal population was assessed with Random Amplified Microsatellites (RAMS. Results of this study indicated that anthracnose in Tahiti lime and tamarillo are caused by Colletotrichun acutatum whereas symptoms on mango were induced by the species Colletotrichum gloeosporioides, which was also fund in few citrus samples. RAMS data analysis indicated the existence of two distinct species groups, with a low similarity index (35%. RAM profiles also showed a clear host differentiation of isolates. The C. acutatum population originated from tamarillo exhibited a narrow and homogeneous genetic base, while the C. acutatum population from Tahiti lime was more heterogeneous and genetically complex, as determined by the analysis of molecular variance (AMOVA and of Ni-Li coefficient. The C. gloeosporioides population originated from mango and Tahiti lime was heterogeneous and highly diverse, with clear host differentiation according to RAM profiles. Collectively, the results from this study provide new insight into the general characteristics of Colletotrichum populations on various hosts; this type of knowledge will prove useful in designing more effective management practices.

  19. Potential aflatoxin and ochratoxin a production by Aspergillus species in poultry feed processing.

    Science.gov (United States)

    Fraga, M E; Curvello, F; Gatti, M J; Cavaglieri, L R; Dalcero, A M; da Rocha Rosa, C A

    2007-04-01

    Poultry feeds are prone to fungal growth and mycotoxin production during processing. The identification of biota with the ability to produce mycotoxins is essential. The aims of this study were (1) to monitor the mycobiota counts at different stages of poultry feed processing; (2) to determine the occurrence of Aspergillus species; (3) to evaluate the natural incidence of aflatoxins and ochratoxin A. The ability of Aspergillus spp. and its teleomorphs isolated here to produce these toxins was also investigated. Samples (144) were collected at random from a factory in Brazil. The occurrence of Aspergillus and Eurotium species was demonstrated on DRBC and DG18 media and the production of aflatoxins and ochratoxin A and their natural incidence were determined by TLC and HPLC methods. A. flavus and E. chevalieri were the most prevalent species isolated. Fungal contamination was not found after the pelleting process, though Aspergillus and Eurotium species were recovered from trough samples. High levels of aflatoxin and ochratoxin A producers were found at all stages of poultry feed processing. Also, high natural contamination with aflatoxins and ochratoxin A was found in the samples. Contact of feed with remainder poultry feed could lead to fungal contamination, so the risk of aflatoxin and/or ochratoxin A contamination of feed must be taken into account.

  20. Antimicrobial activity evaluation and comparison of methods of susceptibility for Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacter spp. isolates.

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    Rechenchoski, Daniele Zendrini; Dambrozio, Angélica Marim Lopes; Vivan, Ana Carolina Polano; Schuroff, Paulo Alfonso; Burgos, Tatiane das Neves; Pelisson, Marsileni; Perugini, Marcia Regina Eches; Vespero, Eliana Carolina

    The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest ® (bioMérieux), Vitek 2 ® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2 ® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  1. Correlation between antibiotic and biocide resistance in mesophilic and psychrotrophic Pseudomonas spp. isolated from slaughterhouse surfaces throughout meat chain production.

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    Lavilla Lerma, Leyre; Benomar, Nabil; Casado Muñoz, María del Carmen; Gálvez, Antonio; Abriouel, Hikmate

    2015-10-01

    The aim of this study was to evaluate biocide susceptibility in mesophilic and psychrotrophic pseudomonads isolated from surfaces of a goat and lamb slaughterhouse, which was representative of the region. To determine biocide resistance in pseudomonads, we determined for the first time the epidemiological cut-off values (ECOFFs) of benzalkonium, cetrimide, chlorhexidine, hexachlorophene, P3 oxonia, polyhexamethylene guanidine hydrochloride (PHMG), topax 66 and triclosan being generally very similar in different Pseudomonas spp. with some exceptions. Thus, resistance of pseudomonads was mainly shown to triclosan, and in lesser extent to cetrimide and benzalkonium chloride depending on the species, however they were highly susceptible to industrial formulations of biocides. By means of statistical analysis, positive correlations between antibiotics, biocides and both antimicrobials in pseudomonads were detected suggesting a co- or cross resistance between different antimicrobials in goat and lamb slaughterhouse environment. Cross-resistance between biocides and antibiotics in pseudomonads were especially detected between PHMG or triclosan and different antibiotics depending on the biocide and the population type. Thus, the use of those biocides as disinfectant in slaughterhouse zones must be carefully evaluated because of the selection pressure effect of antimicrobials on the emergence of resistant bacteria which could be spread to the consumer. It is noteworthy that specific industrial formulations such as topax 66 and oxonia P3 showed few correlations with antibiotics (none or 1-2 antibiotics) which should be taken into consideration for disinfection practices in goat and lamb slaughterhouse. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Incidencia y fármaco-resistencia de cepas de Staphylococcus spp aisladas de exudados conjuntivales Incidence and drug-resistance of Staphylococcus spp strains isolated from conjunctival swabs

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    Andrés Noya Cabaña

    2010-01-01

    Full Text Available OBJETIVO: Determinar la incidencia de cepas del género Staphylococcus aisladas de exudados conjuntivales y analizar su resistencia frente a diferentes antimicrobianos. MÉTODOS: Se realizó un estudio observacional descriptivo retrospectivo en el que se revisaron 3554 exudados conjuntivales a pacientes que acudieron en el período comprendido entre enero de 2002 a diciembre de 2003 y desde enero de 2005 hasta diciembre de 2007 al Instituto Cubano de Oftalmología «Ramón Pando Ferrer» por presentar un diagnóstico de infección ocular externa. RESULTADOS: Se aislaron 874 cepas de Staphylococcus aureus para un 47,5 % y 965 cepas de Staphylococcus spp. coagulasa negativa con prueba de patogenicidad positiva para un 52,4 %. En 69 de esos exudados los cultivos presentaron etiología mixta con dos bacterias diferentes, para el 3,7 %. Se determinaron los porcentajes de resistencia a las cepas aisladas pertenecientes al género Staphylococcus. CONCLUSIONES: Se encontró una alta incidencia de las especies del género Staphylococcus en las infecciones oculares, así como se pudo apreciar que la menor fármaco-resistencia para Staphylococcus aureus y Staphylococcus spp. coagulasa negativa correspondieron a los antimicrobianos ciprofloxacina y amikacina. La mayor fármaco-resistencia de las cepas de Staphylococcus aureus correspondió a eritromicina y tetraciclina y en Staphylococcus spp coagulasa negativa fue frente a la tetraciclina.OBJECTIVE: To determine the incidence of Staphylococcus strains isolated from conjunctival swaps and their resistance to several antimicrobial agents. METHODS: A retrospective, descriptive and observational study was performed to review 3554 conjunctival swabs from patients who went to "Ramón Pando Ferrer" Institute of Ophthalmology in the period from January 2002 to December 2009 due to a diagnosis of external ocular infection. RESULTS: From the total of conjunctival swabs, 874 Staphylococcus aureus strains (47.5 % and

  3. Kinetics of improved 1,4-alpha-D-glucan glucohydrolase biosynthesis from a newly isolated Aspergillus oryzae IIB-6 and parameter significance analysis by 2-factorial design.

    Science.gov (United States)

    Fatima, Bilqees; Ali, Sikander

    2012-01-01

    Sixteen different mould cultures viz. Aspergillus, Alternaria, Arthroderma, Trichoderma, Fusarium, Penicillium, Rhizopus and Chochliobolus were isolated from the soil samples of Qatar by serial dilution method. The preliminary screening of isolates was done by selecting initial colonies showing relatively bigger zones of starch hydrolysis on nutrient agar plates. The isolates were then subjected to secondary screening by submerged fermentation (SmF). The 1,4-α-D-glucan glucohydrolase (GGH) activity ranged from 1.906-12.675 U/ml/min. The product yield was analysed in dependence of mycelial morphology, biomass level and protein content. The isolate Aspergillus oryzae llB-6 which gave maximum enzyme production was incubated in M3 medium containing 20 g/l starch, 10 g/l lactose, 8.5 g/l yeast extract, 6 g/l corn steep liquor (CSL), 1.2 g/l MgSO4.7H2O, 1.3 g/l NH4Cl, 0.6 g/l CaCl2.2H2O, pH 5 at 30±2°C and 200 rpm. On the basis of kinetic variables, notably Qp (0.058±0.01(a) U/g/h), Yp/s (0.308±0.03(ab) U/g) and qp (0.210±0.032(abc) U/g fungal biomass/h), A. oryzae IIB-6 was found to be a hyper producer of GGH (LSD 0.0345) compared to A. kawachii IIB-2. A noticeable enhancement in enzyme activity of over 30% was observed (13.917±1.01 U/ml/min) when the process parameters viz. cultural conditions (pH 5, incubation period 72 h) and nutritional requirements (6 g/l CSL, 9.5 g/l yeast extract, 10 g/l starch, 20 g/l lactose) were further optimized using a 2-factorial Plackett-Burman design. The model terms were found to be highly significant (HS, p≤0.05), indicating the potential utility of the culture (dof~3).

  4. Drug use and antimicrobial resistance among Escherichia coli and Enterococcus spp. isolates from chicken and turkey flocks slaughtered in Quebec, Canada.

    Science.gov (United States)

    Boulianne, Martine; Arsenault, Julie; Daignault, Danielle; Archambault, Marie; Letellier, Ann; Dutil, Lucie

    2016-01-01

    An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in > 90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.

  5. Aspergillus fumigatus-Related Species in Clinical Practice.

    Science.gov (United States)

    Lamoth, Frédéric

    2016-01-01

    Aspergillus fumigatus is the main etiologic agent of invasive aspergillosis (IA). Other Aspergillus species belonging to the section Fumigati (A. fumigatus complex) may occasionally be the cause of IA. These strains are often misidentified, as they cannot be distinguished from A. fumigatus by conventional morphological analysis and sequencing methods. This lack of recognition may have important consequences as these A. fumigatus-related species often display some level of intrinsic resistance to azoles and other antifungal drugs. A. lentulus, A. udagawae, A. viridinutans, and A. thermomutatus (Neosartorya pseudofischeri) have been associated with refractory cases of IA. Microbiologists should be able to suspect the presence of these cryptic species behind a putative A. fumigatus isolate on the basis of some simple characteristics, such as defect in sporulation and/or unusual antifungal susceptibility profile. However, definitive species identification requires specific sequencing analyses of the beta-tubulin or calmodulin genes, which are not available in most laboratories. Multiplex PCR assays or matrix-assisted laser desorption ionization - time-of-flight mass spectrometry (MALDI-TOF MS) gave promising results for rapid and accurate distinction between A. fumigatus and other Aspergillus spp. of the section Fumigati in clinical practice. Improved diagnostic procedures and antifungal susceptibility testing may be helpful for the early detection and management of these particular IA cases.

  6. Aspergillus fumigatus-Related Species in Clinical Practice

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    Frederic eLamoth

    2016-05-01

    Full Text Available Aspergillus fumigatus is the main etiologic agent of invasive aspergillosis (IA. Other Aspergillus species belonging to the section Fumigati (A. fumigatus complex may occasionally be the cause of IA. These strains are often misidentified, as they cannot be distinguished from A. fumigatus by conventional morphological analysis and sequencing methods. This lack of recognition may have important consequences as these A. fumigatus-related species often display some level of intrinsic resistance to azoles and other antifungal drugs. A. lentulus, A. udagawae, A. viridinutans and A. thermomutatus (Neosartorya pseudofischeri have been associated with refractory cases of IA. Microbiologists should be able to suspect the presence of these cryptic species behind a putative A. fumigatus isolate on the basis of some simple characteristics, such as defect in sporulation and/or unusual antifungal susceptibility profile. However, definitive species identification requires specific sequencing analyses of the beta-tubulin or calmodulin genes, which are not available in most laboratories. Multiplex PCR assays or matrix-assisted laser desorption ionization – time-of-flight mass spectrometry (MALDI-TOF MS gave promising results for rapid and accurate distinction between A. fumigatus and other Aspergillus spp. of the section Fumigati in clinical practice. Improved diagnostic procedures and antifungal susceptibility testing may be helpful for the early detection and management of these particular IA cases.

  7. In Vitro Activities of Amphotericin B, Terbinafine, and Azole Drugs against Clinical and Environmental Isolates of Aspergillus terreus Sensu Stricto

    Science.gov (United States)

    Fernández, Mariana S.; Rojas, Florencia D.; Cattana, María E.; Sosa, María de los Ángeles; Iovannitti, Cristina A.; Giusiano, Gustavo E.

    2015-01-01

    The antifungal susceptibilities of 40 clinical and environmental isolates of A. terreus sensu stricto to amphotericin B, terbinafine, itraconazole, and voriconazole were determined in accordance with CLSI document M38-A2. All isolates had itraconazole and voriconazole MICs lower than epidemiologic cutoff values, and 5% of the isolates had amphotericin B MICs higher than epidemiologic cutoff values. Terbinafine showed the lowest MICs. No significant differences were found when MICs of clinical and environmental isolates were compared. PMID:25824228

  8. Typing and virulence factors of food-borne Candida spp. isolates.

    Science.gov (United States)

    Rajkowska, Katarzyna; Kunicka-Styczyńska, Alina

    2018-08-20

    Food-borne yeasts, excluding yeasts used as starter cultures, are commonly considered as food spoilage microorganisms. However, the incidence of non-C. albicans Candida (NCAC) infections has increased considerably over the past two decades. Although 15 Candida species are frequently identified as pathogens, a threat to human from food-borne Candida is poorly recognized. In the present study food-borne NCAC were characterized for the virulence factors, known to be associated with yeast pathogenicity. All food-borne strains in planktonic forms and 89% in biofilm structures represented biotypes established for C. albicans, and 61% demonstrated hemolytic activity. 56-94% of food-borne isolates formed biofilms on glass and biomaterials at a level comparable to clinical C. albicans. Nine out of eighteen tested food-borne NCAC strains (C. krusei, C. lusitaniae, C. famata, C. colliculosa, C. parapsilosis, C. tropicalis) showed similarity to clinical C. albicans in terms of their biotypes and the tested virulence factors, allocating them in a group of risk of potential pathogens. However, their capacity to grow at 37 °C seems to be the preliminary criterion in the study of potential virulence of food-borne yeasts. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. Isolation of Salmonella spp. in cattle egrets (Bubulcus ibis) from Fernando de Noronha Archipelago, Brazil.

    Science.gov (United States)

    Silva, Marcio A; Fernandes, Érika F S T; Santana, Sandra C; Marvulo, Maria Fernanda V; Barros, Mércia R; Vilela, Sineide M O; Reis, Eliane M F; Mota, Rinaldo A; Silva, Jean C R

    2018-03-03

    The growth of the population of cattle egrets (Bubulcus ibis) in the archipelago of Fernando de Noronha constitutes a threat to public health and biological diversity because of their competition with and predation on native species and the possibility of transmission of pathogens to human beings, livestock and native wildlife. The aim here was to search for, isolate and identify serovars of Salmonella in clinically healthy local cattle egrets. Cloacal swabs were obtained from 456 clinically healthy cattle egrets of both sexes and a variety of ages. The swabs were divided into 51 pools. Six of these (11.7%) presented four serovars of Salmonella enterica subspecies enterica: Salmonella serovar Typhimurium; Salmonella serovar Newport; Salmonella serovar Duisburg; and Salmonella serovar Zega. One sample was identified as S. enterica subspecies enterica O16:y:-. Results in this study suggest that cattle egrets may be reservoirs of this agent on Fernando de Noronha and represent a risk to public health and biological diversity. Copyright © 2018. Published by Elsevier Editora Ltda.

  10. Genetic and phenotypic characterization of Saccharomyces spp. strains isolated in distillery plants.

    Science.gov (United States)

    Úbeda, Juan F; Chacón-Ocaña, Maria; Díaz-Hellín, Patricia; Ramírez-Pérez, Hector; Briones, Ana

    2016-06-01

    In this study, the biodiversity and some interesting phenotypic properties of Saccharomyces wild yeasts isolated in distilleries, at least 100 years old, located in La Mancha (Spain), were determined. Strains were genetically characterized by RFLP-mtDNA, which confirmed a great genetic biodiversity with 73% of strains with different mtDNA profiles, highlighting the large variability found in sweet and fermented piquette substrata. The predominant species identified was S. cerevisiae, followed by S. paradoxus and S. bayanus Due to the residual sugar-alcohol extraction process using warm water, a great number of thermophilic Saccharomyces strains with a great cell vitality were found to have potential use as starters in distillery plants. Interesting technological properties such as cell vitality and growth rate at different temperatures were studied. The thermal washing process for the extraction of alcohol and reducing sugars of some raw materials contributes to the presence of Saccharomyces strains with technologically interesting properties, especially in terms of vitality and resistance to high temperatures. Due to the fact that fermentation is spontaneous, the yeast biota of these environments, Saccharomyces and non-Saccharomyces, is very varied so these ecological niches are microbial reserves of undoubted biotechnological interest. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Evaluation of resistance to low pH and bile salts of human Lactobacillus spp. isolates.

    Science.gov (United States)

    Fuochi, Virginia; Petronio, Giulio Petronio; Lissandrello, Edmondo; Furneri, Pio Maria

    2015-09-01

    There are nearly 100 trillion bacteria in the intestine that together form the intestinal microbiota. They are 'good' bacteria because they help to maintain a physiological balance and are called probiotics. Probiotics must have some important characteristics: be safe for humans, be resistant to the low pH in the stomach, as well as bile salts and pancreatic juice. Indeed, their survival is the most important factor, so that they can arrive alive in the intestine and are able to form colonies, at least temporarily. The aim of our study was the evaluation of resistance of Lactobacillus isolates from fecal and oral swabs compared to that found in a commercial product. Seven strains were randomly chosen: L. jensenii, L. gasseri, L. salivarius, L. fermentum, L. rhamnosus, L. crispatus, and L. delbrueckii. We observed a large variability in the results: L. gasseri and L. fermentum were the most resistance to low pH, while only L. gasseri showed the best survival rate to bile salts. Interestingly, the commercial product did not show tolerance to both low pH and bile salts. © The Author(s) 2015.

  12. Evaluation of a modified Cefsulodin-Irgasan-Novobiocin agar f