• Title/Summary/Keyword: phage types

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Phage typing and pulsed-field gel electrophoresis of Salmonella typhimurium and S enteritidis isolated from domestic animals in Gyeongbuk province (경북지역 가축에서 분리된 Salmonella typhimurium과 S enteritidis의 phage typing 및 pulsed-field gel electrophoresis)

  • Kim, Sang-Yun;Lee, Hee-Moo;Kim, Sin;Hong, Hyon-Pyo;Kwon, Heon-Il
    • Korean Journal of Veterinary Service
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    • v.24 no.3
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    • pp.243-253
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    • 2001
  • Forty-five Salmonella typhimurium isolates were encountered 8 phage types in which DT197 and U302 were the predominant types. The DT104 type which was first found from pig in Korea, and was resistant to chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim, tetracycline, gentamicin and nalidixic acid. Twenty-two S enteritidis isolates were encountered 5 phage types in which PT4 were the representative (predominant). S enteritidis isolates were susceptible to all antimicrobial agents. As a result of PFGE analysis for S typhimurium and S enteritidis, PFGE patterns was better than phage typing in discriminating of strains. PFGE patterns were not in accord with phage type even though some strain had the same phage types.

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Phage typing of Staphylococcus aureus Isolates from Poultry Meat in Spain

  • Rosa Capita;Astorga, Maite-Alvarez;Calleja, Carlos-Alonso;Benito Moreno
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.219-225
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    • 2001
  • Phage typing is currently used for typing of Staphylococcus aureus strains beyond the species level in epidemiological studies. A total of 168 Staphylococcus aureus isolates from chicken meat and chicken by-products were phage-typed using the international bacteriophage set for typing Staphylococcus aureus of human origin. One hundred and forty-eight (88.79%) strains were phage-typeable (at least one phage produced 20 or more plaques of lysis). Lysis by phages of group Ⅲ was the mast frequent with 99 (58.93%) sensitive strains. This fact coincides with results of other authors. Twenty-nine different phage patterns were observed and three (95, 75/84 and 6/1030/W57) were most common. One hundred and thirty-two (89.19% of typeable strains) skewed these or indistinguishable (only one phage reaction difference) patterns. Twenty-six out of seventy chicken samples (37.14%) harboured more than one phage type of Staphylococcus aureus. This fact emphasizes the convenience of subtyping several Staphylococcus aureus isolates from the same sample in epidemiological studies. 80% of sausages and hamburgers contained the same Staphylococcus aureus phage types, which wore not found in any of the other food types. This fact suggests a cross contamination during the processing of these foods. Phages 6, 75, 84, 1030 and W57 skewed the greatest activity. None of the Staphylococcus aureus strains were sensitive to phages 47, 81 and 94.

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Proteome analysis between diverse phenotypes of Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium)

  • Shin, Gee-Wook;Cha, In-Seok;Lee, Woo-Won;Nho, Seong-Won;Park, Seong-Bin;Jang, Ho-Bin;Kim, Yong-Hwan;Jung, Tae-Sung
    • Korean Journal of Veterinary Research
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    • v.50 no.4
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    • pp.285-295
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    • 2010
  • Protein expression patterns in Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) strains with diverse phenotypes, such as phage type, antibiotic resistance pattern and plasmid profiles were examined. For detailed analysis of proteins expressed by different S. Typhimurium strains, protein fractions were divided into detergent-rich phase (DP) and aqueous phase (AP) using triton X-114 detergent. The two phases were subjected to two-dimensional gel electrophoresis (2-DE), followed by protein identification using peptide mass fingerprinting (PMF). In the results, PMF showed that DP fractions consisted mainly of outer membrane proteins, whereas the AP fractions included cytosolic proteins. Comparison of 2-DE profiles of DP did not show any distinct protein spots which could be correlated with phage type, antibiotic resistance pattern or plasmid profile. However, comparisons of 2-DE profiles of the AP revealed differences in the protein spots, which could be correlated with the plasmid profile and phage types. Among these protein spots, flagellin was specific for strains containing a 90 kb plasmid. Compared to DT193 phage type, three protein spots in the range of pI 5.0-5.5 and MW 8-15 kDa of AP 2-DE profiles were absent in the DT104 phage types. Additionally, a protein spot with PI in the range of 4.5-5.0 and molecular weight (MW) between 51-69 kDa was specific for phage type DT104, while a protein spot with pI in the range of 4.0-4.8 and MW between 18-20 kDa was specific for DT193 phage type. These protein spots may be useful for discriminating phage types of S. Typhimurium.

Phage types of Salmonella enteritidis isolated from animal and human in Korea (한국의 동물과 사람에서 분리한 Salmonella enteritidis의 phage types)

  • Woo, Yong-ku;Park, Mi-sun;Woo, Seung-ryong;Kim, Bong-hwan;Kim, Jae-hak
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.515-524
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    • 2000
  • The present study was conducted to investigate the phage types(PT) of Salmonella enteritidis(SE) isolated from animals and human patients in Korea during the period 1993-1999. SE were isolated from chickens including the layer and broiler, duck, rat, swine and human patients in nationwide scale. A total of 231 non-human and 14 human SE strains were phage typed and 80.4%(197/245) of SE isolates tested could discriminate as a ten SEPTs including the RDNC by the 10 standard phages, which were brought from the Central Public Health Laboratory in England. In analysing of phage types both animal and human SE isolates, SEPT 4(55.4%) was the most prevalent in domestic animals including the layer(39.3%) and broiler(64.5%), duck(55.5%), and swine(75.0%) since 1993. About 57% of the S enteritidis from human was SEPT 4. Along with SEPT 4, SEPT 1, SEPT 6a, SEPT 7 and SEPT 7a var, SEPT 9b, SEPT 15, SEPT 21b and SEPT 22 were also found from animal and human SE isolates. SEPT 4(40.0%) was isolated from frozen chicken meats imported from China along with SEPT 7 and 7a var strains in this study. In conclusion, SEPT 4 was the most predominant SEPT in our country regardless of animal or human hosts including the Che-Ju island and almost exclusively found in domestic poultry(layer and broiler). During the review period, SEPT 4 has already been spreading continually among the animal hosts since 1993.

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Vi-Phage Type Distribution and Antibiotic Susceptibility of Salmonella typhi in Korean(1982) (장티푸스균이 Vi-phage형 및 항생제감수성에 관한 연구)

  • Lee, B.K.;Kim, B.H.;Chung, T.H.
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.39-46
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    • 1983
  • We collected three hundred thirty-five strains of Salmonella typhi isolated from human sources during the period January to December 1982 Korea. Most of them were from general hospitals and city health center, the remaining one hundred sixtyone strains were obtained from other 12 provincial health centers among 335 testing strains. We used ninety-nine Vi-phages as distributed from International Center for Enteric phage Typing(ICEPT) in London. We found nine phage types among 335 strains of Salmonella typhi in this study. Additionally, I+IV, degraded Vi-positive and Vi negative strains were presented. This study documented the occurrence of the new $B_2$ type in Korea for the first time. The present basic phage type formula in Korea appeared to be A, $B_2,\;D_1,\;D_2,\;D_4,\;D_6,\;D_8,\;D_{12},\;E_1,\;E_9,\;D_1,\;M_1$, 40 and plus I+IV and degraded vi strains(Table 2). The current phage types of Salmonella typhi isolated in Korea 1982 were A, $B_2,\;D_1,\;D_2,\;D_6,\;D_8,\;E_1,\;M_1$ and 46 $M_1$ type was widely distributed all over the country, and E type was next predominant. Antibiotic susceptibility test were performed by means of Kirby-Bauer disc diffution method using 12 kind of antibiotics such as Ampicillin, Carbenicillin, Cephalosporin, Chloramphenicol, Colistin, Gentamicin, Kanamycin, Nalidix acid, Neomycin, Polymyxin-B, Streptomycin and Tetracycline. The sensitivity pattern to antibiotics of Salmonelia typhi cultures were summarized.

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Studies on the Bacteriophages of Brevibacterium lactofermentum (L-글루타민산 생산균 Brevibacterium lactofermentum의 Bacteriophag에 관한 연구)

  • 이태우
    • Korean Journal of Microbiology
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    • v.17 no.3
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    • pp.97-130
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    • 1979
  • Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

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Serotypes, antimicrobial resistance of Salmonella spp. and plasmid profiles, phage types, PFGE of S. Enteritidis and S. Typhimurium isolated from ducks in Daegu-Gyeongbuk province (대구.경북지역 오리 유래 Salmonella속 균의 혈청형, 항균제 내성 및 S. Enteritidis와 S. Typhimurium의 plasmid profiles, phage types 및 PFGE)

  • Cho, Jae-Keun;Kang, Min-Su;Kim, Ki-Seuk
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.217-226
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    • 2011
  • Salmonella spp. is of increasing public health concern as causative pathogens of food poisoning. The aim of this study was to investigate the serotypes and antimicrobial resistance pattern of Salmonella spp. isolated from duck farms in Daegu-Gyeongbuk province. Also, S. Enteritidis and S. Typhimurium isolates were further examined for plasmid analysis, phage typing and pulsed-field gel electrophoresis (PFGE). A total of 34 Salmonella spp. (16.4%) were isolated from duck farms and ten serotypes were identified. The predominant serotypes were S. Typhimurium (23.5%) S. Fyris (17.6%) and S. Haardt (11.8%), S. Agona and S. Enteritidis (respectively 8.8%). Of 34 Salmonella isolates, 15 (44.1%) isolates were resistant to at least one antimicrobial agent and multiple resistance (resistance to more than 4 drugs) was observed in 9 strains (26.5%). The high resistance was found to streptomycin (32.4%), tetracycline (29.4%), ampicillin, kanamycin and nalidixic acid (respectively, 26.5%), all Salmonella isolates were susceptible to cefoxitin, cefotaxime, gentamicin, amikacin and ciprofloxacin. All S. Enteritidis and S. Typhimurium isolates were found to contain only one plasmid (ca. 54 or 55kb, respectively). Among the S. Enteritidis isolates, two phage types were found, PT32a and PT1c, respectively, one isolates did not react with any of the phages used. Whereas, all S. Typhimurium isolates were RDNC (reacts but does not conform). PFGE showed to be a useful typing method better than plasmid analysis and phage typing for discrimination of isolates especially, S. Typhimurium isolates. Our results indicated that the serotypes of Salmonella isolates are widely distributed in duck farms, further epidemiological studies should be carried out.

Effect of Phosphates on Lytic Activity of Bacteriophages Infected in Lactobacilus Cells (유산간균 Bacteriophage의 증식억제물질)

  • 강국희;박기문
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.253-258
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    • 1982
  • Lactobacillus casei YIT 9018 was infected with phage J1 and subjected to grow in $Ca^{++}$ -free MRT (spell out) medium under the presence of four different types of phosphates, sodium-metaphosphate,-pyrophos-phate,-dibasic phosphate, and potassium-phosphate. Among the phosphates tested, sodium pyrophosphate showed sufficient inhibition on the lytic activity of the phage at 0.1% level whereas other phosphate needed more than 0.2% for the same effect. When the concentration of sodium pyrophosphate increased to 0.17%, the bacteria could be protected from lysis until the second succeeding transfer.

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Molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolated from pigs and cattle (소와 돼지유래 다제내성 Salmonella속 균의 분자유전학적 특성)

  • Lee, Woo-Won;Jung, Byeong-Yeal;Lee, Gang-Rok;Lee, Dong-Soo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.32 no.1
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    • pp.61-76
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    • 2009
  • At the present study, it was aimed to explore the molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolates from pigs and cattle. A total of 138 Salmonella Typhimurium (S. Typhimurium) isolates were typed with phage, among them, 83.3% of S. Typhimurium tested could divide into a 10 phage types. Definitive type 193 (DT193) (25.4%) and DT195 (24.6%) were exhibited as the dominant types. DT104 and U302 were found from pigs and cattle. On the other hand, S. Enteritidis had 6 phage types, of them, phage type 21 (PT21) and PT11b were the popular types. In the plasmid profiles, 135 of S. Typhimurium isolates were exhibited 1 to 6 plasmid bands which molecular weight ranged from 90 to 2kb. 35 isolates (25.4%) harbored a 90kb plasmid which is thought to be the serotype specific virulence plasmid. Two of twenty five S. Enteritidis had common plasmids at 2 and 1.5kb. With multiplex polymerase chain reaction, virulence genes (invA and spvC) were detected from all Salmonella spp. from 167 of S. Typhimurium, S. Enteritidis and chloramphenicol resistant S. Schwarzengrund, but some drug resistant genes, such as PSE-1, cml/tetR and flo were not determined but other drug resistant genes, for example TEM and int were found. The detection rates of spvC, TEM and int gene was 35.3%, 29.3% and 72.5%, respectively. The TEM gene was highly popular in S. Typhimurium, which was detected from ampicillin and amoxicillin resistant strains as 95.9%. int gene was able to detect from all the isolates identified as multidrug resistsnt (MDR), particularly DT193 was thought as the most prevalent virulence and multidrug resistance isolate. The major plasmid profile and drug resistance pattern of DT193 were 90, 40, 10.5, 6.3, 3.0kb and ACCbDNaPSSuT, respectively. MDR was commonly found in other phage types, particularly DT104, U302 and DT203.

Genotyping, Phage Typing, and Antimicrobial Resistance of Salmonella Typhimurium Isolated from Pigs, Cattle, and Humans

  • Ju, Min-Seok;Kang, Zheng-Wu;Jung, Ji-Hun;Cho, Seong-Beom;Kim, Sung-Hun;Lee, Young-Ju;Hong, Chong-Hae;Pak, Son-Il;Hahn, Tae-Wook
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.47-53
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    • 2011
  • Salmonella enterica serovar Typhimurium (ST) is one of the most common serovars isolated from humans and animals. It has been suggested that ST infections in Koreans are largely due to the consumption of contaminated pork and beef. To investigate the genotypes, phage types, and antimicrobial resistance patterns for ST isolates of different origins, a total of 70 ST strains, including 19 isolates from humans, 44 isolates from pigs, and 6 isolates from cattle, were analyzed using pulsedfield gel electrophoresis (PFGE), phage typing, and antimicrobial susceptibility tests. Forty-three distinct PFGE patterns were generated from 70 ST isolates, which were grouped into 14 PFGE groups (from A to N) at the level of 75% similarity. The most prevalent group was the A (A1-A17 subtypes) group, encompassing 54.5% (38/70) of ST isolates. ST isolates from pigs and cattle mostly belong to groups A and L, whereas ST isolates from humans mostly belong to groups F and C. Antimicrobial susceptibility tests using 11 antimicrobial agents showed that resistance to tetracycline (TE) (81.4%) was highly prevalent, followed by streptomycin (S) (64.3%) and nalidixic acid (NA) (31.4%) resistance. A total of seventeen antimicrobial resistance patterns were observed. Only 8.6% of isolates, including a reference strain, were susceptible to all antimicrobial agents tested. The most prevalent resistance pattern was TE-S (37.1%), which was seen in 66.6% of bovine, 40.8% of swine and 21.1% of human isolates. Three ST isolates from humans (15.9%) showed resistance to 7-8 antimicrobials. The most predominant phage type (PT) was U302 (64.3%), followed by DT170 (10.0%). PFGE types did not coincide with antimicrobial resistance patterns and phage types; therefore, the combination of those types allowed for further differentiation between tested ST isolates.