Refine search
Results 1-10 of 66
Linkage of serum resistance, aerobactin production, and resistance to antimicrobial agents on conjugal plasmids in some strains of Escherichia coli isolated from septic foals.
1993
Hirsh D.C. | Kirkham C. | Wilson W.D.
Fifteen isolates of Escherichia coli obtained from the blood and tissues of septic foals had plasmid DNA of size ranging from 2.5 to 93 megadaltons. These isolates grew in normal equine serum (serum resistant), a trait previously documented to be expressed by isolates obtained from blood and tissues of septic foals, but not by isolates obtained from the feces of clinically normal horses. Of these isolates, 3 contained conjugal plasmids that encoded resistance to multiple antimicrobial agents linked to serum resistance and, in 1 isolate, to production of aerobactin as well. Serum resistance and production of aerobactin are related to virulence of septicemic E coli from non-equine sources.
Show more [+] Less [-]Association between the effacing (eae) gene and the Shiga-like toxin-encoding genes in Escherichia coli isolates from cattle.
1993
Mainil J.G. | Jacquemin E.R. | Kaeckenbeeck A.E. | Pohl P.H.
Two hundred ninety-six Eschericbia coli isolates from feces or intestines of calves with diarrhea were hybridized with 7 gene probes. One probe (the eae probe) was derived from the eae gene coding for a protein involved in the effacement of the enterocyte microvilli by the group of bacteria called attaching and effacing E coli (AEEC), and 2 probes were derived from genes coding for the Shiga-like toxins (SLT) 1 and 2 produced by the verocytotoxic E coli (VTEC). The other 4 probes were derived from DNA sequences associated with the adhesive properties of enteroadherent E coli (EAEC) to cultured cells (the EAF probe for the localized adherence pattern, probes F1845 and AIDA-1 for the diffuse adherence pattern, and the Agg probe for the aggregative adherence pattern). Hybridization results for the eae probe were in agreement, for all but 1 of the 8 isolates, with previously published phenotypic results of microvilli effacement. The latter was previously reported as effacing the microvilli of calf enterocytes, but was eae probe-negative. Two classes of isolates hybridized with the eae probe. Members of a first class (60 isolates) additionally produced a positive signal with 1 or both of the SLT probes (VTEC-AEEC isolates). Isolates hy- bridizing with the eae and the SLT1 probes were the most frequent: 56 isolates (ie, 93% of all VTEC-AEEC). Members of the second class (10 isolates) failed to hybridize with either SLT probe (non-VTEC-AEEC isolates). Most isolates of these 2 classes belong to only 4 serogroups: O5, O26, O111, and O118. In addition to these 2 AEEC classes, a VTEC class (20 isolates) was observed. Such isolates were positive with 1 or both SLT probes, but were negative with the eae probe. All but 1 isolate belonged to serogroups not found among the AEEC isolates. Only 7 of all AEEC and VTEC isolates were positive with the EAF, the F1845, or the AIDA-1 probe, and none were positive with the Agg probe. On the other hand, 32 non-VTEC, non-AEEC isolates were po.
Show more [+] Less [-]Evaluation of a live avirulent Escherichia coli vaccine for K88+, LT+ enterotoxigenic colibacillosis in weaned pigs.
1991
Francis D.H. | Willgohs J.A.
Live, avirulent Escherichia coli vaccine strains were constructed and tested for efficacy in preventing colibacillosis in 4-week-old pigs. Either or both of 2 plasmids were inserted into avirulent E coli strain G58-1 (0101:NM). These plasmids were pPMC4, which encodes for LTb subunits of heat-labile enterotoxin, and pDHF1, which encodes for K88ac fimbriae. Litter- and weight-matched pigs were removed from sows when they were 10 days old and vaccinated orally with the constructed strains or with G58-1 (negative control vaccine) when they were 2 weeks old and 5 days later. All pigs were challenge-inoculated with virulent E coli strain 3030-2 (0157:K88, LT+, STb+) 2 weeks after the first vaccination. Only 1 pig vaccinated with G58-1/pPMC4/pDHF1 developed diarrhea and none died following challenge inoculation. Seventeen of 31 control pigs developed diarrhea and 11 died. Of 18 pigs vaccinated with G58-1/pDHF1 then challenge-inoculated with the virulent strain, 5 developed diarrhea and 2 died. Fifteen of 18 litter- and weight-matched controls developed diarrhea and 8 died. When compared with G58-1 (negative control), G58-1/pPMC4 afforded no protection to pigs challenge-inoculated with 3030-2.
Show more [+] Less [-]Genetic properties of R plasmids in Salmonella isolates of swine and bovine origin in Korea., 2; incompatibility and profile of R plasmid.
1989
Choi W.P. | Lee H.S. | Yeo S.G. | Lee H.J. | Jung S.C.
This paper deals with the genetic properties of R plasmids in Salmonella originated from pigs and cattle. The plasmid DNA was examined for incompatibility, stability and fertility inhibition (F1), and gel electrophoresis was performed for isolation of plasmid DNA. Among the 66 conjugative R plasmids from 44 pigs and 22 cattle, 61 R plasmids (92.4 %) were Fi-, whereas the remainder were Fi+. The Inc groups of 66 R plasmids were determined with 7 standard plasmids. Twenty-six R plasmids were classified into Inc group Ialpha, H1, H2 or F1, 40 R plasmids being not classified with standard plasmids used, and the Inc group Ialpha (57.7 %) was most frequent. 3. Inc groups Ialpha H1, and F1 were identified in strains from swine, Inc groups H2 and F1 from cattle. The plasmid DNA profiles in 16 Salmonella isolated from pigs and cattle were confirmed as being 1 to 10 fragments by the gel eletrophoresis. Their molecular weight ranged 1.0 to 90 megadalton. The molecular weight of conjugative plasmids ranged 1.0 to 80 megadalton in 4 Salmonella (P-4, P-5, P-7 and P-8) isolated from pigs.
Show more [+] Less [-]Genetic properties of R plasmids in Shigella isolates of swine origin in Korea.
1989
Choi W.P. | Kwon H.B. | Jung S.C.
This paper dealt wih the distribution of Shigella spp. on 5 piggeries in Taegu and Kyongbuk during the period from August to October 1987. Isolated Shigella were examined for serogrouping, antimicrobial drug resistance and detection of R plasmid. Genetic properties of R plasmid in Shigella were examined for fertility inhibition (F1) and gel electrophoresis was performed for the isolation of plasmid DNA. Of total 2,978 samples from 5 piggeries, 82 strains (2.8 %) of Shigella spp. were isolated from 82 samples. The isolated strains were identified as S. dysenteriae (60 strains), S. flexneri (20 strains) and S. sonnei (2 strains). Of the 82 strains examined 67 (95.1 %) were resistant to one or more antibiotics, such as ampicillin (Am), chloramphenicol (Cm), kanamycin (Km), nalidixic acid (Na), rifampicin (Rf), streptomycin (Sm), sulfademethoxine (Su), and tetracycline (Tc) and higher resistance to Su (90.2 %), Sm (63.4 %) and Tc (63.4 %). Of the 78 resistant Shigella strains 26 (33.3 %) harbored conjugative R plasmids and the transfer frequency of Sm (50.0 %), Cm (33.3 %) resistance was much higher than that of the other drug resistance. The most common resistant patterns were SmSuTc, Su and AmSmSuTc. Out of the 26 Shigella R plasmids examined for Fi, 14 (53.8 % were Fi + and the remainder were Fi-. The plasmid DNA profiles in Shigella spp. (9 strains) isolated from pigs were confirmed as being 2 to 9 fragments by the gel electrophoresis. Their molecular size ranged 2.17 to 87.62 kilobase (Kb). All strains of Shigella spp. consisted in 15.4 Kb plasmids.
Show more [+] Less [-]Genetic properties of R plasmids in Salmonella isolates of swine and bovine origin in Korea, 1. Distribution and drug resistance of Salmonella isolated from dairy cow.
1988
Choi W.P. | Lee H.S. | Chae T.C. | Yeo S.G. | Lee H.J.
This paper deals with the distribution of Salmonella (S) infection on 4 herds in Kyungju and Taegu during the period from May to October 1986. Isolated Salmonella were examined for serotypes, antimicrobial drug resistance and detection of R plasmid. From 4 herds, 67 Salmonella were isolated from 51 samples (1.1%), and their serovar strains were S. typhimurium 6, S. derby 5, S. infantis 4, S. bareilly 4, S. dublin 3, S. anatum 2, S. montevideo 2 and untypable 41. In 4 herds, the incidence of drug resistance was 57.7-100% and transfer frequency of conjugative R plasmid was 96.1-100%.
Show more [+] Less [-]Characterisation of Yersinia enterocolitica strains isolated from wildlife in the northwestern Italian Alps Full text
2022
Carella, Emanuele | Romano, Angelo | Domenis, Lorenzo | Robetto, Serena | Spedicato, Raffaella | Guidetti, Cristina | Pitti, Monica | Orusa, Riccardo
Characterisation of Yersinia enterocolitica strains isolated from wildlife in the northwestern Italian Alps Full text
2022
Carella, Emanuele | Romano, Angelo | Domenis, Lorenzo | Robetto, Serena | Spedicato, Raffaella | Guidetti, Cristina | Pitti, Monica | Orusa, Riccardo
Yersiniosis is a zoonosis causing gastroenteritis, diarrhoea, and occasionally reactive arthritis and septicaemia. Cases are often linked to meat consumption and the most common aetiological agent is the Gram-negative bacilliform Yersinia enterocolitica bacterium. The occurrence of Yersinia spp. among wild animals has mostly been studied in wild boar, but it has seldom been in other species. A total of 1,868 faecal samples from animals found dead or hunted were collected between 2015 and 2018 in the Valle d’Aosta region of the northwestern Italian Alps. Alpine ibex faecal samples were collected during a health monitoring program in 2018. Bacteria were isolated via PCR and confirmed as Y. enterocolitica biochemically. Strain antimicrobial susceptibility was tested by Kirby–Bauer disc diffusion, and the presence of virulence factors and antimicrobial resistance genes was investigated using whole-genome sequencing. Yersinia enterocolitica strains of biotype 1A were detected in six faecal samples from red deer (0.93%), roe deer (0.49%) and red foxes (0.7%). Strains found in beech martens (3.57%) and Alpine ibex (2.77%) belonged to biotypes 1B and 5, respectively and harboured the pYPTS01 plasmid that had only been detected in Y. pseudotuberculosis PB1/+. All the isolates were resistant to ampicillin and erythromycin. The biovar 1A strains exhibited different virulence factors and behaved like non-pathogenic commensals. The strain from an Alpine ibex also harboured the self-transmissible pYE854 plasmid that can mobilise itself and the pYPTS01 plasmid to other strains. The beech marten could be considered a sentinel animal for Y. enterocolitica. Phenotypic resistance may account for the ability of all the strains to resist β-lactams.
Show more [+] Less [-]Characterisation of Yersinia enterocolitica strains isolated from wildlife in the northwestern Italian Alps Full text
2022
Carella Emanuele | Romano Angelo | Domenis Lorenzo | Robetto Serena | Spedicato Raffaella | Guidetti Cristina | Pitti Monica | Orusa Riccardo
Yersiniosis is a zoonosis causing gastroenteritis, diarrhoea, and occasionally reactive arthritis and septicaemia. Cases are often linked to meat consumption and the most common aetiological agent is the Gram-negative bacilliform Yersinia enterocolitica bacterium. The occurrence of Yersinia spp. among wild animals has mostly been studied in wild boar, but it has seldom been in other species.
Show more [+] Less [-]Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6) Full text
2019
Wang, Lu-Lu | Lu, Shi-Ying | Hu, Pan | Fu, Bao-Quan | Li, Yan-Song | Zhai, Fei-Fei | Ju, Dan-Di | Zhang, Shi-Jun | Su, Bing | Zhou, Yu | Liu, Zeng-Shan | Ren, Hong-Lin
Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6) Full text
2019
Wang, Lu-Lu | Lu, Shi-Ying | Hu, Pan | Fu, Bao-Quan | Li, Yan-Song | Zhai, Fei-Fei | Ju, Dan-Di | Zhang, Shi-Jun | Su, Bing | Zhou, Yu | Liu, Zeng-Shan | Ren, Hong-Lin
Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of Brucella and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and Brucella infection. Material and Methods: The target open reading frame (ORF) DNAs of Prdx6 with a single active centre were prepared using gene splicing by overlap extension PCR (SOE-PCR), and the recombinant eukaryotic expression plasmids inserted by Prdx6 with the single activity centre were constructed and transfected into murine Raw264.7 macrophages. The glutathione peroxidase activity and phospholipase A2 activity of the constructed Prdx6 were examined. Results: The core centres (Ser³² and Cys⁴⁷) of Prdx6 were successfully mutated by changing the 94ᵗʰ nucleotide from T to G and the 140ᵗʰ nucleotide from G to C in the two enzyme activity cores, respectively. The constructed recombinant plasmids of Prdx6 with the single active centre were transfected into murine macrophages showing the expected single functional enzyme activity, which MJ33 or mercaptosuccinate inhibitors were able to inhibit. Conclusion: The constructed mutants of Prdx6 with the single activity cores will be a benefit to further study of the biological function of Prdx6 with different enzyme activity.
Show more [+] Less [-]Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6) Full text
2019
Wang Lu-Lu | Lu Shi-Ying | Hu Pan | Fu Bao-Quan | Li Yan-Song | Zhai Fei-Fei | Ju Dan-Di | Zhang Shi-Jun | Su Bing | Zhou Yu | Liu Zeng-Shan | Ren Hong-Lin
Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of Brucella and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and Brucella infection.
Show more [+] Less [-]Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China Full text
2016
Hou, Yi-Xuan | Xie, Chun | Wang, Kang | Zhao, Yu-Ting | Xie, Yang-Yang | Shi, Hong-Yan | Chen, Jian-Fei | Feng, Li | Tong, Guang-Zhi | Hua, Xiu-Guo | Yuan, Cong-Li | Zhou, Yan-Jun | Yang, Zhi-Biao
Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China Full text
2016
Hou, Yi-Xuan | Xie, Chun | Wang, Kang | Zhao, Yu-Ting | Xie, Yang-Yang | Shi, Hong-Yan | Chen, Jian-Fei | Feng, Li | Tong, Guang-Zhi | Hua, Xiu-Guo | Yuan, Cong-Li | Zhou, Yan-Jun | Yang, Zhi-Biao
Introduction: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.Material and Methods: Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.Results: The developed quantitative PCR assay detected viral titres as low as 0.1 TCID₅₀ with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive.Conclusion: This approach is suitable for clinical sample identification and pathogenesis studies.
Show more [+] Less [-]Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China Full text
2016
Hou Yi-Xuan | Xie Chun | Wang Kang | Zhao Yu-Ting | Xie Yang-Yang | Shi Hong-Yan | Chen Jian-Fei | Feng Li | Tong Guang-Zhi | Hua Xiu-Guo | Yuan Cong-Li | Zhou Yan-Jun | Yang Zhi-Biao
Introduction: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.
Show more [+] Less [-]Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli Full text
2021
Yu, Hai-Yang | Gao, Dong-Mei | Zhou, Wei | Xia, Bing-Bing | He, Zhi-Yuan | Wu, Bo | Jiang, Min-Zhi | Wang, Mingli | Zhao, Jun
Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli Full text
2021
Yu, Hai-Yang | Gao, Dong-Mei | Zhou, Wei | Xia, Bing-Bing | He, Zhi-Yuan | Wu, Bo | Jiang, Min-Zhi | Wang, Mingli | Zhao, Jun
Ovine interferon-tau (oIFN-τ) is a newly discovered type I interferon. This study used biochemical techniques to transform the oIFN-τ gene into Escherichia coli to obtain the mass and soluble expression of the recombinant protein. First, total RNA was extracted from fresh sheep embryonic tissues with TRIzol reagent and then used as a template to reverse transcribe and amplify the mature oIFN-τ gene with RT-PCR. The amplified product was next digested with the HindIII and XhoI restriction enzymes and inserted into the pET-32a(+) vector to construct the prokaryotic expression plasmid. The corrected in-frame recombinant plasmid, pET-32a(+)-oIFN-τ, was transformed into E. coli Rosetta (DE3) competent cells. After induction with isopropyl-beta-D-thiogalactopyranoside (IPTG), the recombinant protein was detected in bacteria. Finally, the bacteria were lysed by sonication, and the recombinant protein was purified by nickel affinity chromatography and DEAE anion exchange chromatography. The protein was confirmed to be oIFN-τ, which mainly existed in the soluble lysate fraction, as proven by SDS-PAGE and Western blot assays. Purified IFN-τ exists mostly in a soluble form, and its anti-vesicular stomatitis virus (VSV) activity reached 7.08×10(6)IU/mL.
Show more [+] Less [-]Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli Full text
2021
Yu Hai-Yang | Gao Dong-Mei | Zhou Wei | Xia Bing-Bing | He Zhi-Yuan | Wu Bo | Jiang Min-Zhi | Wang Ming-Li | Zhao Jun
Ovine interferon-tau (oIFN-τ) is a newly discovered type I interferon. This study used biochemical techniques to transform the oIFN-τ gene into Escherichia coli to obtain the mass and soluble expression of the recombinant protein.
Show more [+] Less [-]