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Characterization of antimicrobial resistance and application of RFLP for epidemiological monitoring of thermophilic Campylobacter spp. isolated from dogs and humans in Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Jong-Hyun;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.54 no.2
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    • pp.91-99
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    • 2014
  • An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

Genetic and Phenotypic Diversity of Fenitrothion-Degrading Bacteria Isolated from Soils

  • Kim, Kyung-Duk;Ahn, Jae-Hyung;Kim, Tae-Sung;Park, Seong-Chan;Seong, Chi-Nam;Song, Hong-Gyu;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.113-120
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    • 2009
  • Twenty-seven fenitrothion-degrading bacteria were isolated from different soils, and their genetic and phenotypic characteristics were investigated. Analysis of the 16S rDNA sequence showed that the isolates were related to members of the genera Burkholderia, Pseudomonas, Sphingomonas, Cupriavidus, Corynebacterium, and Arthrobacter. Among the 27 isolates, 12 different chromosomal DNA fingerprinting patterns were obtained by polymerase chain reaction(PCR) amplification of repetitive extra genic palindromic(REP) sequences. The isolates were able to utilize fenitrothion as a sole source of carbon and energy, producing 3-methyl-4-nitrophenol as the intermediate metabolite during the complete degradation of fenitrothion. Twenty-two of 27 isolates were able to degrade parathion, methyl-parathion, and p-nitrophenol but only strain BS2 could degrade EPN(O-ethyl-O-p-nitrophenyl phenylphosphorothioate) as a sole source of carbon and energy for growth. Eighteen of the 27 isolates had plasmids. When analyzed with PCR amplification and dot-blotting hybridization using various specific primers targeted to the organophosphorus pesticide hydrolase genes of the previously reported isolates, none of the isolates showed positive signals, suggesting that the corresponding genes of our isolates had no significant sequence homology with those of the previously isolated organophosphate pesticide-degrading bacteria.

Species Profiles and Antimicrobial Resistance of Non-aureus Staphylococci Isolated from Healthy Broilers, Farm Environments, and Farm Workers

  • Ji Heon Park;Gi Yong Lee;Ji Hyun Lim;Geun-Bae Kim;Kun Taek Park;Soo-Jin Yang
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.792-804
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    • 2023
  • Non-aureus staphylococci (NAS), particularly antimicrobial-resistant NAS, have a substantial impact on human and animal health. In the current study, we investigated (1) the species profiles of NAS isolates collected from healthy broilers, farm environments, and farm workers in Korea, (2) the occurrence of antimicrobial-resistant NAS isolates, especially methicillin resistance, and (3) the genetic factors involved in the methicillin and fluoroquinolone resistance. In total, 216 NAS isolates of 16 different species were collected from healthy broilers (n=178), broiler farm environments (n=18), and farm workers (n=20) of 20 different broiler farms. The two most dominant broiler-associated NAS species were Staphylococcus agnetis (23.6%) and Staphylococcus xylosus (22.9%). Six NAS isolates were mecA-positive carrying staphylococcal cassette chromosome mec (SCCmec) II (n=1), SCCmec IV (n=1), SCCmec V (n=2), or nontypeable SCCmec element (n=2). While two mecA-positive Staphylococcus epidermidis isolates from farm workers had SCCmec II and IV, a mecA-positive S. epidermidis isolate from broiler and a Staphylococcus haemolyticus isolate farm environment carried SCCmec V. The occurrence of multidrug resistance was observed in 48.1% (104/216 isolates) of NAS isolates with high resistance rates to β-lactams (>40%) and fusidic acid (59.7%). Fluoroquinolone resistance was confirmed in 59 NAS isolates (27.3%), and diverse mutations in the quinolone resistance determining regions of gyrA, gyrB, parC, and parE were identified. These findings suggest that NAS in broiler farms may have a potential role in the acquisition, amplification, and transmission of antimicrobial resistance.

Re-identification of Colletotrichum acutatum Species Complex in Korea and Their Host Plants

  • Le Dinh Thao;Hyorim Choi;Yunhee Choi;Anbazhagan Mageswari;Daseul Lee;Seung-Beom Hong
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.384-396
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    • 2023
  • Colletotrichum acutatum species complex is one of the most important groups in the genus Colletotrichum with a high species diversity and a wide range of host plants. C. acutatum and related species have been collected from different plants and locations in Korea and deposited into the Korean Agricultural Culture Collection (KACC), National Institute of Agricultural Sciences since the 1990s. These fungal isolates were previously identified based mainly on morphological characteristics, and a limitation of molecular data was provided. To confirm the identification of species, 64 C. acutatum species complex isolates in KACC were used in this study for DNA sequence analyses of six loci: nuclear ribosomal internal transcribed spacers (ITS), betatubulin 2 (TUB2), histone-3 (HIS3), glyceraldehyde3-phosphate dehydrogenase (GAPDH), chitin synthase 1 (CHS-1), and actin (ACT). The molecular analysis revealed that they were identified in six different species of C. fioriniae (24 isolates), C. nymphaeae (21 isolates), C. scovillei (12 isolates), C. chrysanthemi (three isolates), C. lupini (two isolates), and C. godetiae (one isolate), and a novel species candidate. We compared the hosts of KACC isolates with "The List of Plant Diseases in Korea", previous reports in Korea and global reports and found that 23 combinations between hosts and pathogens could be newly reported in Korea after pathogenicity tests, and 12 of these have not been recorded in the world.

Antibacterial Activity of Streptomyces Strains Isolated from Different Regions of Jordan

  • Hala Khyami-Horani;Amal Al-Aboudi;Musa Abu Zarga;Monther Sadder;Halima Othman
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.474-483
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    • 2023
  • Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.

Fatty Acid Profiles of Marine Benthic Microorganisms Isolated from the Continental Slope of Bay of Bengal: A Possible Implications in the Benthic Food Web

  • Das, Surajit;Lyla, P.S.;Khan, S. Ajmal
    • Ocean Science Journal
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    • v.42 no.4
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    • pp.247-254
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    • 2007
  • Marine bacteria, actionmycetes and fungal strains were isolated from continental slope sediment of the Bay of Bengal and studied for fatty acid profile to investigate their involvement in the benthic food-web. Fifteen different saturated and unsaturated fatty acids from bacterial isolates, 14 from actinomycetes and fungal isolates were detected. The total unsaturated fatty acids in bacterial isolates ranged from 11.85 to 37.26%, while the saturated fatty acid ranged between 42.34 and 80.74%. In actinomycetes isolates, total unsaturated fatty acids varied from 27.86 to 38.85% and saturated fatty acids ranged from 35.29 to 51.25%. In fungal isolates unsaturated fatty acids ranged between 44.62 and 65.52% while saturated FA ranged from 20.80 to 46.30%. The higher percentages of unsaturated fatty acids from the microbial isolates are helpful in anticipating the active participation in the benthic food-web of Bay of Bengal.

Fitness of Dicarboximide-Resistant and Sensitive Monilinia fructicola Isolated from Peach in Korea

  • Lim, Tae-Heon;Yi, Jae-Choon;Chang, Tae-Hyun;Byeongjin Cha
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.205-209
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    • 2001
  • Dicarboximide-resistant isolates of Monilinia fructicola grew readily on media amended with dicarbosimide fungicides, and showed cross-resistance to pentachlo-ronitrobenzene (PCNB). The fitness of resistant isolates was inferior to that of sensitive isolates. Mycelial growth on fungicide-free medium was not significantly different between the dicarboximide-resistant and sensitive isolates. The originally high $\textrm{EC}_{50}$ values of the resistant isolate decreased after storage for 16 weeks at $4^{\circ}$. After inoculation with the mixture of spore suspensions of resistant and sensitive isolates, the re-isolation rate of the resistant spores was significantly reduced regardless of the mixing ratio. From the results, it could be concluded that the competitive ability of the resistant isolates is inferior to the sensitive ones.

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Genetic Variability of Sorghum Charcoal Rot Pathogen (Macrophomina phaseolina) Assessed by Random DNA Markers

  • Bashasab, Rajkumar, Fakrudin;Kuruvinashetti, Mahaling S
    • The Plant Pathology Journal
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    • v.23 no.2
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    • pp.45-50
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    • 2007
  • Genetic diversity among selected isolates of Macrophomina phaseolina, a causal agent of charcoal rot (stalk rot) disease in sorghum was studied using PCR-RAPD markers. A set of ten isolates, from ten different rabi sorghum genotypes representing two traditional sorghum growing situations viz., Dharwad- a transitional high rainfall region and Bijapur- a semi-arid low rainfall region in South India. From a set of 40 random primers tested, amplicon profiles of 15 were reproducible. A total of 149 amplicon levels, with an average of 9.9 bands per primer, were available for analysis, of which 148 were polymorphic (99.3%). It was possible to discriminate all the isolates with any of the 15 primers employed. UPGMA clustering of data indicated that the isolates shared varied levels of genetic similarity within a range of 0.14 to 0.72 similarity coefficient index and it was suggestive that grouping of isolates was not related to sampling location in anyway. A high level of genetic heterogeneity of 0.28 was recorded among the isolates.

Pathogenic effects of porcine reproductive and respiratory syndrome virus isolates in swine tracheal ring culture

  • Park, Bong-kyun;Collins, James E.;Goyal, Sagar M.;Pijoan, Carlos;Joo, Han-soo
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.311-317
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    • 1999
  • Pathogenic effects of 29 different porcine reproductive and respiratory syndrome(PRRS) virus isolates were investigated in swine tracheal ring(STR) cultures by examining their effects on the ciliary activity of STR. Inhibition of ciliary movement and destruction of the tracheal epithelium were seen between 72 and 96 hours postinoculation(PI). Virus replication was demonstrated by examining viral infectivity of the supernatants from the STR cultures. PRRS virus antigen in macrophages was detected by a streptavidin-biotin complex(ABC) immunoperoxidase method. Of the 29 PRRS virus isolates, 8 isolates were classified into pathogenic, and the remaining 21 isolates were determined as mildly pathogenic or apathogenic viruses. These results suggest that STR examination may be used as a method for predicting pathogenic variability of PRRS virus isolates.

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Isolation and characterization of Brachyspira pilosicoli in Korean dogs (국내의 개에서 Brachyspira pilosicoli의 분리 및 동정)

  • Kim, Tae-Jung;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.421-426
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    • 2004
  • This study was performed to report the outbreaks of canine intestinal spirochetosis and to characterize the canine isolates. Three canine isolates were weakly beta-hemolytic and had sharp end shape with 5 flagella. Isolates didn't produce indole but fermented fructose. In $API-ZYM^{(R)}$ study, isolates were alpha-glucosidase negative and alpha-galactosidase positive, which is the typical characteristics of B. pilosicoli. In multilocus enzyme electrophoresis(MEE) study, isolates were divided into 2 electrophoretic types. Isolates showed different properties with B. hyodysenteriae but closely related with pathogenic canine intestinal spirochete(B. pilosicoli). This is the first report of the isolation and characterization of canine intestinal spirochete in Korean dogs.