scholarly journals Isolation, molecular identification and antimicrobial resistance patterns of Campylobacter species of dairy origin: first report from Bangladesh

2019 ◽  
Vol 8 (1) ◽  
Author(s):  
S. M. Lutful Kabir ◽  
Most. Mostary Lubna ◽  
Mehedul Islam ◽  
A.K.M. Ziaul Haque ◽  
Sucharit Basu Neogi ◽  
...  
Keyword(s):  
Dairy Farm ◽  
Multidrug Resistant ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Isolation And Identification ◽  
First Report ◽  
Multiplex Pcr Assay ◽  
Faecal Samples ◽  
Resistance Patterns ◽  
Campylobacter Species

This study was aimed for isolation, identification and characterization of Campylobacter species from Bangladesh Agricultural University (BAU) dairy farm during the period of January to May, 2016. A total of 80 samples (faecal samples of calves, heifers and cows; milk samples of cows) were collected from Bangladesh Agricultural University dairy farm for isolation and identification of Campylobacter species by using cultural, biochemical and molecular methods. Moreover, the isolated Campylobacter species were subjected for antimicrobial susceptibility test. Campylobacter like organisms were presumptively identified in 20 samples. Isolates were biochemically positive to catalase and oxidase tests and in hippurate hydrolysis test some of the isolates (n=6) shown negative that indicated the isolates were C. coli and some of the test isolates (n=14) shown positive that indicated the isolates were C. jejuni. Campylobacter specific 16S rRNA genes were amplified from the isolates. Out of 20 isolated Campylobacter 14 (17.5%) were detected as C. jejuni and the rest 6 (7.5%) were detected as C. coli by cdtC gene based multiplex PCR assay. C. jejuni were resistant to amoxicillin, erythromycin, azithromycin and susceptible to gentamicin, ciprofloxacin, norfloxacin and streptomycin. Furthermore, C. coli were resistant to amoxicillin and erythromycin and susceptible to gentamycin, ciproflaxacin. Out of 20 Campylobacter isolates, 57.14% C. jejuni and 33.33% C. coli were identified as multidrug resistant. To the best of our knowledge, this study has brought the first report on the occurrence of Campylobacter species with their antibiogram profiles in any dairy farm of Bangladesh.

scholarly journals Isolation and Characterization of Novel Marine-Derived Actinomycete Taxa Rich in Bioactive Metabolites

2004 ◽  
Vol 70 (12) ◽  
pp. 7520-7529 ◽  
Author(s):  
Nathan A. Magarvey ◽  
Jessica M. Keller ◽  
Valerie Bernan ◽  
Martin Dworkin ◽  
David H. Sherman
Keyword(s):  
16S Rrna ◽  
Multidrug Resistant ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Bioactive Metabolites ◽  
Rrna Gene ◽  
New Genera ◽  
Isolation And Identification ◽  
Fermentation Products ◽  
Isolation And Characterization

ABSTRACT A unique selective enrichment procedure has resulted in the isolation and identification of two new genera of marine-derived actinobacteria. Approximately 90% of the microorganisms cultured by using the presented method were from the prospective new genera, a result indicative of its high selectivity. In this study, 102 actinomycetes were isolated from subtidal marine sediments collected from the Bismarck Sea and the Solomon Sea off the coast of Papua New Guinea. A combination of physiological parameters, chemotaxonomic characteristics, distinguishing 16S rRNA gene sequences, and phylogenetic analysis based on 16S rRNA genes provided strong evidence for the two new genera (represented by strains of the PNG1 clade and strain UMM518) within the family Micromonosporaceae. Biological activity testing of fermentation products from the new marine-derived actinomycetes revealed that several had activities against multidrug-resistant gram-positive pathogens, malignant cells, and vaccinia virus replication.

scholarly journals Prevalence and molecular detection of fluoroquinolone-resistant genes (qnrA and qnrS) in Escherichia coli isolated from healthy broiler chickens

2018 ◽  
pp. 1720-1724 ◽  
Author(s):  
Shahin Mahmud ◽  
K. H. M. Nazmul Hussain Nazir ◽  
Md. Tanvir Rahman
Keyword(s):  
Escherichia Coli ◽  
Broiler Chickens ◽  
Molecular Detection ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Diffusion Method ◽  
Isolation And Identification ◽  
E Coli ◽  
Pcr Targeting ◽  
Apparently Healthy

Aim: The present study was carried out to determine the prevalence and molecular detection of fluoroquinolone-resistant Escherichia coli carrying qnrA and qnrS genes in healthy broiler chickens in Mymensingh, Bangladesh, and also to identify the genes responsible for such resistance. Materials and Methods: A total of 65 cloacal swabs were collected from apparently healthy chickens of 0-14 days (n=23) and 15-35 days (n=42) old. The samples were cultured onto Eosin Methylene Blue Agar, and the isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties followed by polymerase chain reaction (PCR) targeting E. coli 16S rRNA genes. The isolates were subjected to antimicrobial susceptibility test against five commonly used antibiotics under fluoroquinolone (quinolone) group, namely gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, and pefloxacin by disk diffusion method. Detection of qnrA and qnrS genes was performed by PCR. Results: Among the 65 cloacal samples, 54 (83.08%) were found to be positive for E. coli. Antibiotic sensitivity test revealed that, of these 54 isolates, 18 (33.33%) were found to be resistant to at least one fluoroquinolone antibiotic. The highest resistance was observed against pefloxacin (61.11%). By PCR, of 18 E. coli resistant to fluoroquinolone, 13 (72.22%) were found to be positive for the presence of qnrS. None of the isolates were found positive for qnrA. Conclusion: Fluoroquinolone-resistant E. coli harboring qnrS genes is highly prevalent in apparently healthy broiler chickens and possesses a potential threat to human health.

scholarly journals First Report of Meloidogyne marylandi Infecting Bermudagrass in Oklahoma

Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1286-1286 ◽  
Author(s):  
N. Walker
Keyword(s):  
Cynodon Dactylon ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Similar Species ◽  
Root Knot Nematode ◽  
Putting Greens ◽  
Egg Masses ◽  
First Report ◽  
Mean Width ◽  
Ultradwarf Bermudagrass

Meloidogyne marylandi is a nematode commonly associated with turfgrasses and has been reported to occur in Texas and Arkansas (1,3). In the fall of 2013, a stand of ultradwarf bermudagrass (Cynodon dactylon × C. transvaalensis) plants in a sand-based, research putting green in Stillwater, Oklahoma, exhibited symptoms of decline. Roots of the affected plants had small galls and upon staining of the root system, numerous egg masses were evident. Egg masses were collected, placed in water, and the morphology of 20 hatched, second-stage juveniles were examined. The characteristics of the juveniles were: body length averaged 393.1 ± 19.87 (range: 361 to 425) μm, mean width averaged 16.6 ± 0.7 (15.6 to 17.8) μm, stylet lengths averaged 12.1 ± 0.7 (10.4 to 12.9) μm, dorsal gland orifice from stylet base averaged 2.9 ± 0.4 (2.5 to 3.6) μm, tail lengths averaged 53.7 ± 3.8 (46.2 to 60.4) μm, and the hyaline region of the tails averaged 10.4 ± 1.1 (8.4 to 12.7) μm. Genomic DNA was extracted from six females that were removed from roots. Amplification and sequencing of the mitochondrial DNA region between COII and 16S rRNA genes was performed with primers 1RNAF (5′-TACCTTTGACCAATCACGCT-3′) and CO11R (5′-GGTCAATGTTCAGAAATTTGTGG-3′) as previously described (2). A PCR product approximately 510 bp in length was obtained and sequenced at the Oklahoma State University Core Facility. Sequences were compared with those in NCBI's nucleotide database using BLAST and had 97% identity with two sequences from M. marylandi (KC473862.1 and KC473863.1) and the next most similar species being M. graminis (JN241898.1) with 83% identity. To our knowledge, this is the first report of the root-knot nematode M. marylandi in Oklahoma. As bermudagrass becomes more commonly used for putting greens in the turfgrass transition zone, M. marylandi may become a more common and damaging pathogen in the region. References: (1) A. A. Elmi et al. Grass For. Sci. 55:166, 2000. (2) M. A. McClure et al. Plant Dis. 96:635, 2012. (3) J. L. Starr et al. Nematrop. 37:43, 2007.

Development of multiplex PCR assay for concurrent detection of tick borne haemoparasitic infections in bovines

2018 ◽  
Vol 63 (4) ◽  
pp. 759-765 ◽  
Author(s):  
V.R. Kundave ◽  
Hira Ram ◽  
Partha S. Banerjee ◽  
Rajat Garg ◽  
K. Mahendran ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genomic Dna ◽  
Large Scale ◽  
Simultaneous Detection ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Individual Species ◽  
Pcr Assay ◽  
Blood Samples ◽  
Multiplex Pcr Assay

Abstract This study describes development and evaluation of a multiplex PCR assay for simultaneous detection of Theileria annulata, Babesia bigemina and Anaplasma marginale infections in bovines. The assay was developed using parasites specific genomic DNA and three sets of PCR primers targeting the Tams1, 18S rRNA and 16S rRNA genes of T. annulata, B. bigemina and A. marginale, respectively. Blood samples collected from a total of 461 bovines, suspected for haemoparasitic infections, were examined microscopically to record the status of infection and simultaneously, genomic DNA extracted from these blood samples were utilized for the optimization and validation of multiplex PCR assay. Microscopic examination of blood samples revealed presence of single and multiple species of haemoparasites in 25.8% and 2.4% samples, respectively. Results of multiplex PCR revealed the presence of single haemoparasitic species infection in 159 cases (34.5%), whereas mixed infection was recorded in 82 (17.8%) samples. Occurrence of individual species infection detected by mPCR in the study was 26.03% (120/461) for T. annulata, 3.25% (15/461) for B. bigemina and 5.20% (24/461) for A. marginale. The detection limit of multiplex PCR assay was at the template dilutions of 10−6, 10−6 and 10−4, which corresponded to 0.1 pg, 0.1 pg and 10.0 pg of DNA for T. annulata, A. marginale, and B. bigemina, respectively. Based on the high diagnostic sensitivity and throughput, multiplex PCR assay developed in the present study could be exploited as a tool to conduct large-scale epidemiological survey for tick-borne haemoparasitic infection of bovines.

scholarly journals Simultaneous Detection of Aeromonas salmonicida, Flavobacterium psychrophilum, and Yersinia ruckeri, Three Major Fish Pathogens, by Multiplex PCR

2002 ◽  
Vol 68 (10) ◽  
pp. 5177-5180 ◽  
Author(s):  
A. del Cerro ◽  
I. Marquez ◽  
J. A. Guijarro
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Aeromonas Salmonicida ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Yersinia Ruckeri ◽  
Pcr Assay ◽  
Fish Pathogens ◽  
Flavobacterium Psychrophilum ◽  
Multiplex Pcr Assay

ABSTRACT A multiplex PCR assay based on the 16S rRNA genes was developed for the simultaneous detection of three major fish pathogens, Aeromonas salmonicida, Flavobacterium psychrophilum, and Yersinia ruckeri. The assay proved to be specific and as sensitive as each single PCR assay, with detection limits in the range of 6, 0.6, and 27 CFU for A. salmonicida, F. psychrophilum, and Y. ruckeri, respectively. The assay was useful for the detection of the bacteria in artificially infected fish as well as in fish farm outbreaks. Results revealed that this multiplex PCR system permits a specific, sensitive, reproducible, and rapid method for the routine laboratory diagnosis of infections produced by these three bacteria.

scholarly journals Root-Knot Nematodes in Golf Course Greens of the Western United States

Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 635-647 ◽  
Author(s):  
Michael A. McClure ◽  
Claudia Nischwitz ◽  
Andrea M. Skantar ◽  
Mark E. Schmitt ◽  
Sergei A. Subbotin
Keyword(s):  
United States ◽  
Western Hemisphere ◽  
Western United States ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Golf Course ◽  
28S Rrna ◽  
Golf Courses ◽  
Root Knot Nematodes ◽  
First Report

A survey of 238 golf courses in 10 states of the western United States found root-knot nematodes (Meloidogyne spp.) in 60% of the putting greens sampled. Sequence and phylogenetic analyses of 18S rRNA, D2-D3 of 28S rRNA, internal transcribed spacer-rRNA, and mitochondrial DNA gene sequences were used to identify specimens from 110 golf courses. The most common species, Meloidogyne naasi, was found in 58 golf courses distributed from Southern California to Washington in the coastal or cooler areas of those states. In the warmer regions of the Southwest, M. marylandi was recovered from 38 golf courses and M. graminis from 11 golf courses. This constitutes the first report of M. marylandi in Arizona, California, Hawaii, Nevada, and Utah, and the first report of M. graminis in Arizona, Hawaii, and Nevada. Two golf courses in Washington were infested with M. minor, the first record of this nematode in the Western Hemisphere. Columbia root-knot nematode, M. chitwoodi, was found in a single golf course in California. Polymerase chain reaction restriction fragment length polymorphism of the intergenic region between the cytochrome oxidase and 16S rRNA genes in the mitochondrial genome with restriction enzyme SspI was able to distinguish populations of M. graminis from M. marylandi, providing a fast and inexpensive method for future diagnosis of these nematodes from turf.

Phylogenetic relationships and diversity of β-rhizobia associated with Mimosa species grown in Sishuangbanna, China

2011 ◽  
Vol 61 (2) ◽  
pp. 334-342 ◽  
Author(s):  
Xiao Yun Liu ◽  
Wei Wu ◽  
En Tao Wang ◽  
Bin Zhang ◽  
Jomo Macdermott ◽  
...  
Keyword(s):  
16S Rrna ◽  
Phylogenetic Relationships ◽  
Phenotypic Diversity ◽  
Mainland China ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Whole Cell ◽  
First Report ◽  
Sds Page

In order to investigate the genetic diversity of rhizobia associated with various exotic and invasive species in tropical mainland China, 116 bacterial isolates were obtained from Mimosa root nodules collected from Sishuangbanna and Yuanjiang districts of Yunnan province. Isolated rhizobia were characterized by RFLP analysis of 16S rRNA genes, SDS-PAGE of whole-cell proteins and BOX-PCR. Most of the isolated strains were identified as β-rhizobia belonging to diverse populations of Burkholderia and Cupriavidus, and the phylogenetic relationships of their 16S rRNA gene sequences showed that they were closely related to one of four β-rhizobia species: Burkholderia phymatum, B. mimosarum, B. caribensis or Cupriavidus taiwanensis. Additionally, among the 116 isolates, 53 different whole-cell SDS-PAGE profiles and 30 distinct BOX-PCR genotypic patterns were detected, which demonstrated the genetic and phenotypic diversity found within these Burkholderia and Cupriavidus strains. To the best of our knowledge, this is the first report that β-rhizobia are extant and possibly widespread on the Chinese mainland and nodulate easily with Mimosa plants. We also find it especially interesting that this appears to be the first report from mainland China of Cupriavidus symbionts of Mimosa. These records enrich our knowledge and understanding of the geographical distribution and diversity of these bacteria.

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