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Edy M Vilei - One of the best experts on this subject based on the ideXlab platform.
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detection of mycoplasma mycoides subsp mycoides sc in bronchoalveolar lavage fluids of cows based on a taqman real time pcr discriminating wild type Strains from an lppq mutant Vaccine Strain used for diva strategies
Journal of Microbiological Methods, 2010Co-Authors: Edy M VileiAbstract:Contagious bovine pleuropneumonia (CBPP) is the most serious cattle disease in Africa, caused by Mycoplasma mycoides subsp. mycoides small-colony type (SC). CBPP control strategies currently rely on vaccination with a Vaccine based on live attenuated Strains of the organism. Recently, an lppQ− mutant of the existing Vaccine Strain T1/44 has been developed (Janis et al., 2008). This T1lppQ− mutant Strain is devoid of lipoprotein LppQ, a potential virulence attribute of M. mycoides subsp. mycoides SC. It is designated as a potential live DIVA (Differentiating Infected from Vaccinated Animals) Vaccine Strain allowing both serological and etiological differentiation. The present paper reports on the validation of a control strategy for CBPP in cattle, whereby a TaqMan real-time PCR based on the lppQ gene has been developed for the direct detection of M. mycoides subsp. mycoides SC in ex vivo bronchoalveolar lavage fluids of cows and for the discrimination of wild type Strains from the lppQ− mutant Vaccine Strain.
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contagious bovine pleuropneumonia cbpp caused by Vaccine Strain t1 44 of mycoplasma mycoides subsp mycoides sc
Veterinary Microbiology, 2004Co-Authors: Rosastella Mbulu, Edy M Vilei, Paola Pilo, Robin A J Nicholas, Georgina Tjipurazaire, Rosella Lelli, Felix Mettler, Otto J B HuebschleAbstract:A study was carried out on four adult cattle to assess the pathogenicity of Mycoplasma mycoides subsp. mycoides SC Strain T1/44, currently used as a Vaccine for the control of contagious bovine pleuropneumonia (CBPP) in Namibia. Post mortem examination 9 weeks after endobronchial inoculation of the Vaccine Strain to three of the four animals revealed unilateral pleuropneumonic lesions, pleuritis and well-developed sequesters in two of the three inoculated animals and several small sequesters surrounded by pleuropneumonic lesions in the diaphragmatic and apical lobes in one animal. The fourth animal, which was not directly inoculated but was in close contact with the inoculated animals, revealed only an adhesion area of the lung to the ribcage. Serological examination carried out using the complement fixation test (CFT) detected positive titres in all three intubated animals and the indirect CBPP-LppQ-ELISA was positive for two of the three inoculated animals. The contact animal showed no seroconversion. M. mycoides subsp. mycoides SC was isolated from the sequesters of two of the inoculated animals. Isolation of mycoplasmas was not possible from the third inoculated animal due to heavy contamination of the samples by other bacteria, but the presence of M. mycoides subsp. mycoides SC could be evidenced by PCR from clinical samples. The identity of the T1/44 Vaccine Strain isolated from the sequesters of two animals was confirmed by T1/44-specific PCR analysis and by IS1296 typing using Southern blot. These results clearly show that inoculation of T1/44 Vaccine via the endobronchial route can lead to CBPP.
Vito G Delvecchio - One of the best experts on this subject based on the ideXlab platform.
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comparative proteome analysis of brucella melitensis Vaccine Strain rev 1 and a virulent Strain 16m
Journal of Bacteriology, 2002Co-Authors: Michel Eschenbrenner, Mary Ann Wagner, Troy A Horn, Jo Ann Kraycer, Cesar V Mujer, Sue D Hagius, Philip H Elzer, Vito G DelvecchioAbstract:The genus Brucella consists of bacterial pathogens that cause brucellosis, a major zoonotic disease characterized by undulant fever and neurological disorders in humans. Among the different Brucella species, Brucella melitensis is considered the most virulent. Despite successful use in animals, the Vaccine Strains remain infectious for humans. To understand the mechanism of virulence in B. melitensis, the proteome of Vaccine Strain Rev 1 was analyzed by two-dimensional gel electrophoresis and compared to that of virulent Strain 16M. The two Strains were grown under identical laboratory conditions. Computer-assisted analysis of the two B. melitensis proteomes revealed proteins expressed in either 16M or Rev 1, as well as up- or down-regulation of proteins specific for each of these Strains. These proteins were identified by peptide mass fingerprinting. It was found that certain metabolic pathways may be deregulated in Rev 1. Expression of an immunogenic 31-kDa outer membrane protein, proteins utilized for iron acquisition, and those that play a role in sugar binding, lipid degradation, and amino acid binding was altered in Rev 1.
Miklos Gyuranecz - One of the best experts on this subject based on the ideXlab platform.
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development of mismatch amplification mutation assays for the differentiation of ms1 Vaccine Strain from wild type mycoplasma synoviae and ms h Vaccine Strains
PLOS ONE, 2017Co-Authors: Zsuzsa Kreizinger, Kinga M Sulyok, Denes Grozner, Katinka Bekő, Zoltan Szabo, Miklos GyuraneczAbstract:Mycoplasma synoviae is an economically significant pathogen in the poultry industry, inducing respiratory disease and infectious synovitis in chickens and turkeys, and eggshell apex abnormality in chickens. Eradication, medication and vaccination are the options for controlling M. synoviae infection. Currently there are two commercial, live, attenuated Vaccines available against M. synoviae: the temperature sensitive MS-H Vaccine Strain and the NAD independent MS1 Vaccine Strain. Differentiation of Vaccine Strains from field isolates is essential during vaccination and eradication programs. The present study provides melt-curve and agarose gel based mismatch amplification mutation assays (MAMA) to discriminate the MS1 Vaccine Strain from the MS-H Vaccine Strain and wild-type M. synoviae isolates. The assays are based on the A/C single nucleotide polymorphism at nt11 of a HIT family protein coding gene. The melt- and agarose-MAMAs reliably distinguish the MS1 Vaccine Strain genotype from the MS-H Vaccine Strain and wild-type M. synoviae isolate genotype from 102 template number/DNA sample. No cross-reactions with other avian Mycoplasma species were observed. The assays can be performed directly on clinical samples and they can be run simultaneously with the previously described MAMAs designed for the discrimination of the MS-H Vaccine Strain. The developed assays are applicable in laboratories with limited facilities and promote the rapid, simple and cost effective differentiation of the MS1 Vaccine Strain.
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rapid simple and cost effective molecular method to differentiate the temperature sensitive ts ms h Vaccine Strain and wild type mycoplasma synoviae isolates
PLOS ONE, 2015Co-Authors: Zsuzsa Kreizinger, Kinga M Sulyok, Alexandra Pasztor, Karoly Erdelyi, Orsolya Felde, Janos Povazsan, Laszlo Kőrosi, Miklos GyuraneczAbstract:Mycoplasma synoviae infection in chickens and turkeys can cause respiratory disease, infectious synovitis and eggshell apex abnormality; thus it is an economically important pathogen. Control of M. synoviae infection comprises eradication, medication or vaccination. The differentiation of the temperature sensitive (ts+) MS-H Vaccine Strain from field isolates is crucial during vaccination programs. Melt-curve and agarose gel based mismatch amplification mutation assays (MAMA) are provided in the present study to distinguish between the ts+ MS-H Vaccine Strain, its non-temperature sensitive re-isolates and wild-type M. synoviae isolates based on the single nucleotide polymorphisms at nt367 and nt629 of the obg gene. The two melt-MAMAs and the two agarose-MAMAs clearly distinguish the ts+ MS-H Vaccine Strain genotype from its non-temperature sensitive re-isolate genotype and wild-type M. synoviae isolate genotype, and no cross-reactions with other Mycoplasma species infecting birds occur. The sensitivity of the melt-MAMAs and agarose-MAMAs was 103 and 104 copy numbers, respectively. The assays can be performed directly on clinical samples and they can be run simultaneously at the same annealing temperature. The assays can be performed in laboratories with limited facilities, using basic real-time PCR machine or conventional thermocycler coupled with agarose gel electrophoresis. The advantages of the described assays compared with previously used methods are simplicity, sufficient sensitivity, time and cost effectiveness and specificity.
Otto J B Huebschle - One of the best experts on this subject based on the ideXlab platform.
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contagious bovine pleuropneumonia cbpp caused by Vaccine Strain t1 44 of mycoplasma mycoides subsp mycoides sc
Veterinary Microbiology, 2004Co-Authors: Rosastella Mbulu, Edy M Vilei, Paola Pilo, Robin A J Nicholas, Georgina Tjipurazaire, Rosella Lelli, Felix Mettler, Otto J B HuebschleAbstract:A study was carried out on four adult cattle to assess the pathogenicity of Mycoplasma mycoides subsp. mycoides SC Strain T1/44, currently used as a Vaccine for the control of contagious bovine pleuropneumonia (CBPP) in Namibia. Post mortem examination 9 weeks after endobronchial inoculation of the Vaccine Strain to three of the four animals revealed unilateral pleuropneumonic lesions, pleuritis and well-developed sequesters in two of the three inoculated animals and several small sequesters surrounded by pleuropneumonic lesions in the diaphragmatic and apical lobes in one animal. The fourth animal, which was not directly inoculated but was in close contact with the inoculated animals, revealed only an adhesion area of the lung to the ribcage. Serological examination carried out using the complement fixation test (CFT) detected positive titres in all three intubated animals and the indirect CBPP-LppQ-ELISA was positive for two of the three inoculated animals. The contact animal showed no seroconversion. M. mycoides subsp. mycoides SC was isolated from the sequesters of two of the inoculated animals. Isolation of mycoplasmas was not possible from the third inoculated animal due to heavy contamination of the samples by other bacteria, but the presence of M. mycoides subsp. mycoides SC could be evidenced by PCR from clinical samples. The identity of the T1/44 Vaccine Strain isolated from the sequesters of two animals was confirmed by T1/44-specific PCR analysis and by IS1296 typing using Southern blot. These results clearly show that inoculation of T1/44 Vaccine via the endobronchial route can lead to CBPP.
Menachem Banai - One of the best experts on this subject based on the ideXlab platform.
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identification of the brucella melitensis Vaccine Strain rev 1 in animals and humans in israel by pcr analysis of the psti site polymorphism of its omp2 gene
Journal of Clinical Microbiology, 2002Co-Authors: Svetlana Bardenstein, Michal Mandelboim, Thomas A Ficht, Miriam Baum, Menachem BanaiAbstract:Adverse effects of Strain persistence and secretion in milk have been encountered with the Brucella melitensis Vaccine Strain Rev.1. Field isolates obtained from vaccinated animals and from a human resembled the Vaccine Strain Rev.1 by conventional bacteriological tests. The lack of a specific molecular marker that could specifically characterize the commercial Vaccine Strain prevented confirmation of the homology of the Rev.1-like field isolates to the Vaccine Strain. The composition of the omp2 locus from two gene copies with differences in their PstI restriction endonuclease sites was used to establish an epidemiologic fingerprint for the omp2 gene in the Rev.1 Vaccine Strain. Primers designed to amplify DNA sequences that overlap the PstI site revealed a single 282-bp DNA band common to all Brucella spp. Agarose gel electrophoresis of the PstI digests of the PCR products from Strains 16M and the Vaccine Strain Rev.1 revealed a distinctive profile that included three bands: one band for the intact 282-bp fragment amplified from omp2a and two bands resulting from the digestion of the amplified omp2b gene fragment, 238- and 44-bp DNA fragments, respectively. Amplified fragments of 37 Rev.1-like isolates, including 2 human isolates, also exhibited this pattern. In contrast, DNA digests of all other Israeli field isolates, including atypical B. melitensis biotype 1 and representatives of the biotype 2 and 3 isolates, produced two bands of 238 and 44 bp, respectively, corresponding with the digestion of both omp2a and omp2b genes. This method facilitates identification of the Rev.1 Vaccine Strain in both animals and humans in Israel.
