Detection and Characterization of a Multi-drug and Multi-metal Resistant Enterobacterium Pantoea sp. from Tannery Wastewater after Secondary Treatment Process

2016 ◽  
Vol 2 (1 and 2) ◽  
pp. 37-42 ◽  
Author(s):  
Ashutosh Yadav ◽  
Abhay Raj ◽  
Ram Naresh Bharagava

In this study, an enterobacterium was isolated from treated tannery wastewater and characterized as gram-negative, rod shaped, motile, and lactose fermenting bacterium. Further, based on the 16S rRNA gene sequence analysis, the bacterium was identified as Pantoea sp. with accession number The antibiotic and heavy KJ576899. metal resistant property of isolated bacterium was investigated by the disk diffusion method on Muller-Hinton and nutrient agar medium amended with the increasing concentrations of various toxic metal ions, respectively. Results showed that the isolated bacterium was sensitive for amikacin, ampicillin, cefepime, cetriaxone, chloramphenicol, levofloxacin, meropenem, nalidixic acid, piperacillin and tobramycin, resistant for aztreonam, carbenicillin, cefotaxime, ceftazidime, ciprofloxacin, cotrimoxazole, imepenam, moxifloxacin, streptomycin and tetracycline, but intermediate for amoxicillin and gentamicin. In addition, the bacterium also showed the Minimum Inhibitory Concentration (MIC) values of 250, 500, 160, 190, 600, 700, 500, 380, 600 and 350 μg ml-1 forCu, Cr, Cd, Co, Zn, Fe, Ni, Pb, Mo and As, respectively with a plasmid DNA of 33.5 kb. This multi-drug and multi-metal resistant bacterium can be used as a potential agent for the bioremediation of metal contaminated sites.

2020 ◽  
Vol 36 (1) ◽  
Author(s):  
Estibaliz Sansinenea ◽  
Jessica Vaca ◽  
Norma Elena Rojas ◽  
Candelario Vázquez

A highly potent secondary metabolites-producing Bacillus strain was isolated from Mexican soil (Puebla State), together with other fifty strains. The fifty-one strains were subjected for metabolites extraction and evaluated as antibacterial against several bacteria. The active metabolites of these strains were extracted using amberlite XAD16 absorbent resin. The antibacterial activity of crude extracts of all strains was performed by disk diffusion method against some pathogenic gram positive and gram-negative bacteria. Among all Bacillus strains tested, the most potent strain ELI149 (NRB) was selected for molecular characterization. The nucleotide sequence of the 16S rRNA gene (1.5 Kb) of this strain evidenced a 94% similarity with Bacillus amyloliquefaciens strain IIHR-Ba-2, which showed the highest inhibition against the most bacteria probed even greater inhibition than the standard antibiotic. In conclusion, secondary metabolites extracted from Bacillus amyloliquefaciens strain are highly potent as antibiotic against the most bacteria probed. Identification of which metabolites extracted from amberlite are the responsible of the bacteria growth inhibition will be a challenge.


2013 ◽  
Vol 63 (Pt_5) ◽  
pp. 1834-1839 ◽  
Author(s):  
Elena Ramos ◽  
Martha-Helena Ramírez-Bahena ◽  
Angel Valverde ◽  
Encarna Velázquez ◽  
Doris Zúñiga ◽  
...  

During a study of the ‘tunta’ (frozen-dry potato) production process in Peru, a bacterial strain, LMT03T, was isolated from the straw grass in which the potatoes are dried. This strain was classified into the genus Pseudomonas on the basis of the 16S rRNA gene sequence analysis, and is most closely related to Pseudomonas argentinensis CH01T with 99.3 % identity in this gene and 96 %, 92 % and 86 % identities in rpoB, rpoD and gyrB genes, respectively. Strain LMT03T has a single polar flagellum, like other related yellow-pigment-producing pseudomonads. The major quinone is Q-9. The major fatty acids are C18 : 1ω7c in summed feature 8 (40.82 %), C16 : 1ω6c/C16 : 1ω6c in summed feature 3 (23.72 %) and C16 : 0 (15.20 %). The strain produces oxidase but it does not produce gelatinase, indole, urease, arginine dihydrolase or β-galactosidase. Catalase production was very weak after 28 and 48 h incubation on nutrient agar medium. Nitrate reduction is negative. It does not hydrolyse aesculin. The DNA G+C content is 57.8 mol%. DNA–DNA hybridization results showed lower than 52 % relatedness with respect to the type strain of P. argentinensis , CH01T. These results, together with other phenotypic characteristics, support the definition of a novel species within the genus Pseudomonas , for which the name Pseudomonas punonensis sp. nov. is proposed. The type strain is LMT03T ( = LMG 26839T = CECT 8089T).


2004 ◽  
Vol 54 (4) ◽  
pp. 1055-1061 ◽  
Author(s):  
Carole Feurer ◽  
Dominique Clermont ◽  
François Bimet ◽  
Adina Candréa ◽  
Mary Jackson ◽  
...  

Nine unidentified Gram-positive, lipophilic corynebacteria were isolated from clinical and food samples and subjected to a polyphasic taxonomic analysis. The bacteria were distinguished from Corynebacterium species with validly published names by biochemical tests, fatty acid content and whole-cell protein analysis. Comparative 16S rRNA gene sequence analysis demonstrated unambiguously that the nine strains were related phylogenetically to the species ‘Corynebacterium tuberculostearicum’ and represented a distinct subline within the genus Corynebacterium. On the basis of both phenotypic and phylogenetic evidence, the formal description of Corynebacterium tuberculostearicum sp. nov. is proposed. The type strain of C. tuberculostearicum is Medalle XT (=LDC-20T=CIP 107291T=CCUG 45418T=ATCC 35529T).


2020 ◽  
Vol 70 (9) ◽  
pp. 5032-5039 ◽  
Author(s):  
Jae-Chan Lee ◽  
Kyung-Sook Whang

A Gram-stain-positive actinobacterial strain, designated ANK073T, was isolated from rhizosphere soil sampled at a spinach farming field in Shinan, Republic of Korea. Cells of strain ANK073T were found to be aerobic, non-motile, non-spore-forming rods which could grow at 20–40 °C (optimum, 30 °C), at pH 6.0–10.0 (optimum, pH 6.5–7.5) and at salinities of 0–4 % (w/v) NaCl (optimum, 0 % NaCl). The 16S rRNA gene sequence analysis showed that strain ANK073T belongs to the genus Agromyces with high sequence similarities to Agromyces humatus CD5T (98.8 %), Agromyces tardus SJ-23T (98.5 %) and Agromyces iriomotensis IY07-20T (98.4 %). The phylogenetic analysis indicated that strain ANK073T formed a distinct phyletic line in the genus Agromyces and the results of DNA–DNA relatedness and phylogenomic analysis based on whole genome sequences demonstrated that strain ANK073T could be separated from its closest relatives in the genus Agromyces . The strain contained 2,4-diaminobutylic acid, glycine, d-glutamic acid and d-alanine in the peptidoglycan. The predominant menaquinones were identified as MK-12 and MK-11, and the major fatty acids were anteiso-C17 : 0, anteiso-C15 :  0 and iso-C15:0. The major polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The G+C content of the genome was determined to be 70.2 mol%. On the basis of its phenotypic and chemotaxonomic properties and the results of phylogenetic and phylogenomic analyses, strain ANK073T is considered to represent a novel species in the genus Agromyces , for which the name Agromyces humi sp. nov. is proposed. The type strain is ANK073T (=KACC 18683T=NBRC 111825T).


2018 ◽  
Author(s):  
Christian Vinueza-Burgos ◽  
David Ortega-Paredes ◽  
Cristian Narváez ◽  
Lieven De Zutter ◽  
Jeannete Zurita

AbstractAntimicrobial resistance (AR) is a worldwide concern. Up to a 160% increase in antibiotic usage in food animals is expected in Latin American countries. The poultry industry is an increasingly important segment of food production and contributor to AR. The objective of this study was to evaluate the prevalence, AR patterns and the characterization of relevant resistance genes in Extended Spectrum β-lactamases (ESBL) and AmpC E. coli from large poultry farms in Ecuador. Sampling was performed from June 2013 to July 2014 in 6 slaughterhouses that slaughter broilers from 115 farms totaling 384 flocks. Each sample of collected caeca was streaked onto TBX agar supplemented with cefotaxime (3 mg/l). In total, 176 isolates were analyzed for antimicrobial resistance patterns by the disk diffusion method and for blaCTX-M, blaTEM, blaCMY, blaSHV, blaKPC, and mcr-1 by PCR and sequencing. ESBL and AmpC E. coli were found in 362 flocks (94.3%) from 112 farms (97.4%). We found that 98.3% of the isolates were multi-resistant to antibiotics. Low resistance was observed for ertapenem and nitrofurantoin. The most prevalent ESBL genes were the blaCTX-M (90.9%) blaCTX-M-65, blaCTX-M-55 and blaCTX-M-3 alleles. Most of the AmpC strains presented the blaCMY-2 gene. Three isolates showed the mcr-1 gene. Poultry production systems represent a hotspot for antimicrobial resistance in Ecuador, possibly mediated by the extensive use of antibiotics. Monitoring this sector in national and regional plans of antimicrobial resistance surveillance should therefore be considered.


2020 ◽  
Vol 70 (3) ◽  
pp. 1868-1875 ◽  
Author(s):  
Shan-Hui Li ◽  
Jaeho Song ◽  
Yeonjung Lim ◽  
Yochan Joung ◽  
Ilnam Kang ◽  
...  

A Gram-stain-negative, rod-shaped, aerobic, non-flagellated, chemoheterotrophic bacterium, designated IMCC14385T, was isolated from surface seawater of the East Sea, Republic of Korea. The 16S rRNA gene sequence analysis indicated that IMCC14385T represented a member of the genus Halioglobus sharing 94.6–97.8 % similarities with species of the genus. Whole-genome sequencing of IMCC14385T revealed a genome size of 4.3 Mbp and DNA G+C content of 56.7 mol%. The genome of IMCC14385T shared an average nucleotide identity of 76.6 % and digital DNA–DNA hybridization value of 21.6 % with the genome of Halioglobus japonicus KCTC 23429T. The genome encoded the complete poly-β-hydroxybutyrate biosynthesis pathway. The strain contained summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and C17 : 1 ω8c as the predominant cellular fatty acids as well as ubiquinone-8 (Q-8) as the respiratory quinone. The polar lipids detected in the strain were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, five unidentified phospholipids, an unidentified aminolipid, an unidentified aminophospholipid and four unidentified lipids. On the basis of taxonomic data obtained in this study, it is suggested that IMCC14385T represents a novel species of the genus Halioglobus , for which the name Halioglobus maricola sp. nov. is proposed. The type strain is IMCC14385T (=KCTC 72520T=NBRC 114072T).


2017 ◽  
Vol 84 (3) ◽  
Author(s):  
Irene Cano ◽  
Ronny van Aerle ◽  
Stuart Ross ◽  
David W. Verner-Jeffreys ◽  
Richard K. Paley ◽  
...  

ABSTRACTOne of the fastest growing fisheries in the UK is the king scallop (Pecten maximusL.), also currently rated as the second most valuable fishery. Mass mortality events in scallops have been reported worldwide, often with the causative agent(s) remaining uncharacterized. In May 2013 and 2014, two mass mortality events affecting king scallops were recorded in the Lyme Bay marine protected area (MPA) in Southwest England. Histopathological examination showed gill epithelial tissues infected with intracellular microcolonies (IMCs) of bacteria resemblingRickettsia-like organisms (RLOs), often with bacteria released in vascular spaces. Large colonies were associated with cellular and tissue disruption of the gills. Ultrastructural examination confirmed the intracellular location of these organisms in affected epithelial cells. The 16S rRNA gene sequences of the putative IMCs obtained from infected king scallop gill samples, collected from both mortality events, were identical and had a 99.4% identity to 16S rRNA gene sequences obtained from “CandidatusEndonucleobacter bathymodioli” and 95% withEndozoicomonasspecies.In situhybridization assays using 16S rRNA gene probes confirmed the presence of the sequenced IMC gene in the gill tissues. Additional DNA sequences of the bacterium were obtained using high-throughput (Illumina) sequencing, and bioinformatic analysis identified over 1,000 genes with high similarity to protein sequences fromEndozoicomonasspp. (ranging from 77 to 87% identity). Specific PCR assays were developed and applied to screen for the presence of IMC 16S rRNA gene sequences in king scallop gill tissues collected at the Lyme Bay MPA during 2015 and 2016. There was 100% prevalence of the IMCs in these gill tissues, and the 16S rRNA gene sequences identified were identical to the sequence found during the previous mortality event.IMPORTANCEMolluscan mass mortalities associated with IMCs have been reported worldwide for many years; however, apart from histological and ultrastructural characterization, characterization of the etiological agents is limited. In the present work, we provide detailed molecular characterization of anEndozoicomonas-like organism (ELO) associated with an important commercial scallop species.


2020 ◽  
Vol 367 (4) ◽  
Author(s):  
Seza Arslan ◽  
Fatma Özdemir

ABSTRACT A wide variety of foods can be contaminated with Listeria species, especially L. monocytogenes. Ready-to-eat (RTE) foods are predominantly associated with human listeriosis caused by L. monocytogenes. Therefore, this study aimed to assess the presence of Listeria species in RTE foods and to characterize L. monocytogenes isolates by means of detection of virulence markers, serotypes and genetic relatedness. Of the 300 RTE food samples, 59 (19.7%) were positive for Listeria species: L. innocua (13.3%), L. monocytogenes (5%), L. welshimerii (2.3%), L. grayi subsp. murrayi (1.3%), L. grayi (1%), L. ivanovii (1%) and L. ivanovi subsp. londoniensis (0.3%). All L. monocytogenes isolates identified were positive for the actA, iap, inlA, inlB, inlC, inlJ, plcA and prfA virulence genes and biofilm. The isolates were serotyped as 1/2c (33.3%), 4b (26.7%), 1/2a (26.7%), 1/2b (6.7%) and 3c (6.7%) by the multiplex-PCR and agglutination methods. PCR-restriction fragment length polymorphism with AluI and MluCI resulted in three and two profiles, respectively. Pulsed-field gel electrophoresis differentiated the L. monocytogenes isolates into 15 ApaI and 12 AscI patterns. Antimicrobial resistance of all Listeria isolates was determined by the disk diffusion method. Most L. monocytogenes isolates were sensitive to antimicrobials used in the treatment of listeriosis. This study shows the presence of potential pathogenic and antimicrobial-resistant L. monocytogenes in RTE foods that may lead to consumer health risks.


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