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1.
Salmonella livingstone is one of the more common salmonella serotypes isolated in the United Kingdom. The characterization of 70 different isolates of S. livingstone using biochemical tests and plasmid profile analysis is described. The isolates could be divided into four groups by their ability to grow in d-tartrate, l-tartrate and Stern's glycerol. Further subdivision was achieved by the use of plasmid profile analysis; 24 of the isolates possessed light plasmids (less than 9.5 MD) and four possessed heavy plasmids (greater than 30 MD). The combination of biotyping and plasmid profile analysis can be used as the basis of a scheme to 'fingerprint' S. livingstone for epidemiological studies.  相似文献   

2.
Phenotypic and molecular techniques, including antimicrobial susceptibility testing, plasmid analysis, and pulsed-field gel electrophoresis (PFGE) were used to characterize 15 isolates of multidrug-resistant (MDR) Salmonella anatum cultured during a 16 mo period from horses and a veterinary clinic environment. The isolates were resistant to multiple antimicrobial agents and could be placed into 4 groups based on their antimicrobial resistance patterns. The isolates contained multiple plasmids ranging in size from 2 to > 100 kb that could be grouped into 3 different plasmid profile patterns; these patterns did not correlate with the antimicrobial resistance groupings. Furthermore, antimicrobial resistance was conjugatively transferable. Digestion of genomic DNA from the 15 isolates with 3 different restriction endonucleases, SfiI, SpeI, and XbaI followed by PFGE revealed a highly conserved restriction endonuclease digestion pattern. In contrast, diverse banding patterns were observed with S. anatum obtained from other sources. These observations suggest that the MDR S. anatum isolates represent a common outbreak strain even though they possess different, albeit similar, antibiograms and plasmid profiles. The study showed that PFGE is a useful epidemiological tool for discriminating between unrelated and outbreak-related strains of S. anatum. In conclusion, epidemiological studies of outbreaks caused by MDR isolates of S. anatum should consist of both genotypic and phenotypic methods of analysis.  相似文献   

3.
The aim of the present study was to evaluate the importance of clonal spread of Brachyspira hyodysenteriae resistant to pleuromutilins (tiamulin, valnemulin) on farms in the Czech Republic. Agar dilution method and macrorestriction fragment profile analysis by pulsed field gel electrophoresis were used to characterise 35 B hyodysenteriae isolates that were obtained from clinical cases of swine dysentery on 32 farms between 2000 and 2005. Most isolates showed multiple resistances to tiamulin, valnemulin, tylosin and lincomycin. A total of six pulsotypes were detected in these multiresistant isolates. An analysis of epidemiological data showed that multiresistant B hyodysenteriae isolates were more often detected on fattening farms (59 per cent), compared with farms with other types of production. Furthermore, it was found that multiresistant B hyodysenteriae clones were most frequently selected on farms with endemic incidence of swine dysentery. This finding was confirmed by the characterisation of 21 B hyodysenteriae isolates obtained from three large-scale operations in seven consecutive years.  相似文献   

4.
Restriction deoxyribonucleic acid (DNA) fragment profile analysis coupled with immunogenic protein profile analysis has provided useful information in determining the differences between vaccine strains and field isolates of fowlpox virus (FPV). The DNA of strains examined in this study clearly fell into 3 minor groups of restriction patterns similar but distinct from one another: restriction patterns exhibited by the vaccine strains except 1 vaccine strain, Vac-82; restriction profiles indicated by Vac-82 and field isolates FI-38 and FI-42; and restriction patterns indicated by field isolates FI-43, FI-51, FI-54, and FI-56. Furthermore, when the strains were analyzed and compared by immunoblotting analysis, they showed group differences similar to the differences in restriction profiles. Both techniques provided high sensitivity in verifying differences between vaccine strains and field isolates of FPV. The disparity found in restriction fragments or immunogenic protein profile between vaccine strains and field isolates does not exclude the appreciable high degree of DNA sequence conservation and homology. However, the minor disparity observed in these strains suggests a molecular basis for why vaccinated commercial flocks could have continually been infected by variant strains of FPV. A rapid and sensitive polymerase chain reaction method, which amplified a product from the 4b core protein gene of the FPV genome, was developed for identification and differentiation of members of the genus Avipoxvirus. Whereas total DNA from either vaccine strains or field isolates was used as template for amplifying a predicted product of 578 or 1409 bp, only cleavage of the amplified product (1409 bp) represented an additional detection technique for species differentiation. An attempt to distinguish between strains on the basis of amplification product was partially successful.  相似文献   

5.
The multiple-locus VNTR analysis (MLVA) assay is a method frequently employed as a molecular epidemiological tool for Brucella genetic fingerprinting. The purpose of this study was to assess the genotyping of 77 B. canis isolates from 14 different dog breeding farms in Korea by the MLVA assay and to compare the epidemiological relationships between the Korean isolates and foreign ones. Simpson's diversity index for 17 loci showed a range from 0 to 0.846 in 77 B. canis isolates. B. canis isolates in Korea were observed to have high genetic diversity at the most variable loci and were divided into 30 distinct genotypes by phylogenetic analysis. Some B. canis isolates were closely related to previously typed isolates in other countries. The MLVA assay can be helpful to analyze the epidemiological correlation of B. canis isolates in domestic pet animals and to track the geographic origin by comparing the genetic patterns with foreign isolates. Therefore, the MLVA assay will be useful as a tool for control and preventive measures of canine brucellosis.  相似文献   

6.
The genetic similarity of Campylobacter jejuni isolates from pets, compared to human clinical cases and retail food isolates collected in Ireland over 2001-2006 was investigated by cluster analysis of pulsed-field gel electrophoresis (PFGE) fingerprinting profiles. Comparison of the PFGE profiles of 60 pet isolates and 109 human isolates revealed that seven (4.1%) profiles were grouped in clusters including at least one human and one pet C. jejuni isolate. In total six (1.6%) of 60 pet and 310 food profiles were in clusters with at least one food and one pet C. jejuni isolate. The detection of only a small number of genetically indistinguishable isolates by PFGE profile cluster analysis from pets and from humans with enteritis in this study suggests that pets are unlikely to be an important reservoir for human campylobacteriosis in Ireland. However, genetically indistinguishable isolates were detected and C. jejuni from pets may circulate and may contribute to clinical infections in humans. In addition, contaminated food fed to pets may be a potential source of Campylobacter infection in pets, which may subsequently pose a risk to humans.  相似文献   

7.
From 2005 to 2007, Mycobacterium tuberculosis complex (MTC) strains were isolated from cattle, goats and pigs samples collected at the Bodija abattoir and from human samples from tuberculosis patients and livestock traders at the Akinyele cattle market in Ibadan, Southwestern Nigeria. Seventy four isolates obtained from humans (24) and livestock (50) were identified as MTC strains. Thirty two isolates were spoligotyped. Nineteen of these 32 isolates were identified as M. tuberculosis whilst 13 were identified as Mycobacterium bovis. M. bovis was isolated from two humans, whereas M. tuberculosis was isolated from a bovine, a pig and a goat. All the M. bovis isolates identified in this study belonged to the Africa 1 clonal complex. Multiple locus VNTR [variable number of tandem repeats] analysis (MLVA) was carried out on the 74 isolates. Three major clusters were defined. Group A consisted of 24 M. tuberculosis isolates (MLVA genotypes 1-18). One strain was isolated from a bovine and one from a pig. Group B consisted of 49 M. bovis strains (MLVA genotypes 19-48), mainly of cattle origin but also included four goat, nine pig and two human isolates. Group C consisted of a single M. tuberculosis isolate (MLVA genotype 49) obtained from a goat. Spoligotyping and MLVA confirmed it as clustering with the East Africa Indian clade found in humans in Sudan and the Republic of Djibouti. The isolation of three M. tuberculosis strains from livestock raises the question of their epidemiological importance as a source of infection for humans.  相似文献   

8.
Restriction endonuclease analysis (REA) and plasmid profile were used to study the epidemiology of Pasteurella multocida in a swine pyramid structure. The studied pyramid was comprised of a group of 12 swine farrow-to-finish farms related by unidirectional animal movement. P. multocida isolates were obtained from the lungs of 275 slaughtered pigs. Serotyping was performed by hyaluronidase sensitivity test and toxicity was investigated by the ELISA test. HpaII was used to cleave the P. multocida extracted DNA. REA patterns relationships were studied using the Sokal-Michener coefficients, and the dendrogram was built using the UPGMA system. The 218 P. multocida isolates obtained were distributed in 17 REA patterns. In 9 of the 12 farms studied only 2-3 REA patterns were detected, with one clearly predominant pattern. The 81 strains with plasmids were assigned to six plasmid profiles. REA and plasmid profiles proved to be good epidemiological tools for identifying different strains of P. multocida with the same phenotype.  相似文献   

9.
The banding profiles generated by Bam H1 restriction endonuclease cleavage of bacterial DNA from clinical and reference isolates of Histophilus ovis, Haemophilus somnus and related bacteria were compared. H. ovis, H. somnus and Haemophilus agni isolates were found to have distinct similarities in banding profiles characterised by 10 common bands between 2.0 and 9.6 kilobases (kb). The close taxonomic relationship of these isolates was reinforced by these findings. The reference isolates examined in this study--Actinobacillus lignieresii, Actinobacillus seminis, H. agni, H. somnus, H. ovis, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus parahaemolyticus--could be distinguished from each other on the basis of their characteristic banding profiles. Actinobacillus sp were observed to have more bands between 2 and 23 kb compared with the H. ovis and Haemophilus sp isolates studied. Analysis of isolates from an experimental infection trial illustrated the potential of restriction endonuclease analysis in molecular epidemiological applications. It was possible to demonstrate by this means that the post-challenge isolates had identical banding profiles to the challenge (or infecting) isolate which had a distinctly different banding profile from that of pre-challenge H. ovis isolates. Furthermore, restriction endonuclease analysis of H. ovis isolates obtained from follow-up investigations of a recurrent problem of epididymitis in unmated rams, indicated that the H. ovis isolates implicated in epididymitis, were present as a single strain in a number of sheep over a period of time. This suggested that the mechanism of transmission was by perinatal perputial contamination.  相似文献   

10.
Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate a total of 38 Pasteurella multocida isolates from four separate outbreaks of pasteurellosis in Australian piggeries. Six isolates were obtained from Outbreak 1, 16 from Outbreak 2 and eight each from outbreaks 3 and 4. Outbreaks 1 and 2 were cases of pneumonic pasteurellosis while outbreaks 3 and 4 involved systemic pasteurellosis. Biochemical characterisation established that a number of different types of P. multocida were present in outbreaks 1 and 3 while outbreaks 2 and 4 were associated with a single type of P. multocida. Outbreaks 1 and 3 yielded isolates of P. multocida that belonged to the subspecies multocida and gallicida, with the subspecies multocida isolates being identified as biovar 3 (6 in total) or 12 (1 in total) and the subspecies gallicida isolates (7 in total) being identified as biovar 8. All 24 isolates from outbreaks 2 and 4 belonged to the subspecies multocida and were all biovar 3. REA and ribotyping showed that, in outbreaks 1 and 3, there were three different types of P. multocida in each outbreak with no common strains between the outbreaks. The molecular methods showed that only a single strain of P. multocida was associated with outbreaks 2 and 4, although the outbreaks were associated with strains that differed in REA profiles but shared a ribotype profile. This study has shown that both, systemic and pneumonic pasteurellosis can be associated with either a single strain or multiple strains of P. multocida. The results also indicate that the molecular typing methods of REA and ribotyping are superior to biochemical characterisation for epidemiological investigation of porcine pasteurellosis.  相似文献   

11.
A total of 95 isolates of Pasteurella multocida were analysed by pulsed field gel electrophoresis (PFGE) using the enzyme ApaI, including 73 avian isolates from Australia and 22 from Vietnam. The majority of field isolates were capsular Type A, with the predominant somatic serovars of 1, 3, 4 and 3,4. Twenty-one distinct profiles were evident among the Australian isolates, with only 3 profiles observed among the 22 P. multocida strains isolated from Vietnam. Within the Australian isolates, related and unrelated outbreaks could be identified by PFGE. These results correlated well with previously published studies, with greater discrimination shown by PFGE. Repetitive extragenic palindromic sequence PCR (REP-PCR) analysis of representative isolates from PFGE classifications yielded 21 profiles, with most of the subgroups in accordance with PFGE analysis. While REP-PCR was shown to be less discriminating than PFGE, the epidemiological relatedness of strains compared favourably between the techniques. Thus, the ease and rapidity of REP-PCR while maintaining a high level of differentiation, supports the use of REP-PCR as a competent alternative to the more labour-intensive PFGE system for strain identification and epidemiological studies of avian P. multocida.  相似文献   

12.
In order to differentiate infectious bursal disease virus (IBDV) isolates/strains, a quick method of RT-PCR followed by restriction enzyme analysis of VP1 gene sequence is being reported for the first time. A 480 bp fragment, comprising one of the RNA dependent RNA polymerase motifs of VP1 gene sequence of an Indian classical virus, an attenuated vaccine strain, Georgia and two Indian field isolates, genetically similar to reported very virulent strains of IBDV, was amplified by RT-PCR. Restriction enzyme digestion of PCR products with Taq1 enzyme generated distinct profile for field isolates, different from the classical and attenuated viruses, whereas restriction profile with BstNI restriction enzyme was similar in all the viruses, irrespective of the pathotype. Therefore, the present results suggest that Taq1 digestion can be taken up for the differentiation of field isolates from the classical and vaccine strains. The sequence analysis of VPI gene of reported very virulent IBD viruses from Europe and Japan, using 'MapDraw' programme of Lasergene software, revealed similar restriction enzyme profile as in Indian field isolates.  相似文献   

13.
Bacteroides nodosus involved in several outbreaks of ovine footrot over a number of years were subjected to DNA restriction endonuclease analysis. Individual isolates were found to have characteristic Bam HI profiles which permitted their accurate identification and differentiation from other isolates. Bam HI profiles of B. nodosus isolates were used in epidemiological investigations involving consecutive outbreaks of footrot on individual and neighbouring farms. The relationship of given isolates to a common source could be established by this means. Restriction endonuclease analysis provides an additional epidemiological tool in ovine footrot investigations as it accurately identifies interstrain differences in a manner not possible by conventional bacteriological and serological means.  相似文献   

14.
In 1991 EMCV was isolated for the first time in Belgium from the offspring of a sow with reproductive failure. From August 1995 until December 1996, EMCV was diagnosed in 154 Belgian pig holdings in association with myocardial failure and sudden death in fatteners and suckling piglets or with reproductive failure in sows. To clarify some epidemiological aspects 3 EMCV isolates characteristic for the different clinical pictures and outbreaks were studied. Field observations and animal experiments indicated that the pathogenicity induced by each isolate is specific for one age category and that the spread of the virus is limited. The presented data also suggest that rodents may play a role in the transmission of EMCV but that pig-to-pig transmission is at least as important. Molecular analysis of two separate regions on the genomes of the respective EMCV isolates showed that the 1995-96 EMCV epizootic in Belgium was due to a new virus introduction. Furthermore, the VP1 coding gene is proposed as a marker of virulence.  相似文献   

15.
Since differences in the virulence of Mycoplasma (M.) hyopneumoniae strains have been described, the isolation of field strains followed by genotypic and phenotypic characterisation has become a major goal in epidemiological studies. The aim of this study was to compare various M. hyopneumoniae isolates from different pig herds and numerous pigs within the same herd. Therefore, pigs of 109 herds located in North-Western Germany were sampled either on-farm or during necropsies. Overall, 52 isolates of M. hyopneumoniae were recovered from 45 pigs originating from 21 herds. The identity of cultures was confirmed by PCR targeting the 16S-23S intergenic spacer region. Typing of isolates was achieved by random amplified polymorphic DNA (RAPD) analysis and multi-locus analysis of variable number of tandem repeats (VNTR) and demonstrated a high degree of heterogeneity of M. hyopneumoniae isolates. Differences among isolates recovered from animals of the same herd or even from the same pig revealed a grouping into different genotypic clusters. This outcome was observed with both methods. It was concluded that more than one strain of M. hyopneumoniae might be present in a pig herd and even in a single pig, suggesting high genetic heterogeneity between isolates of the same epidemiological source. These factors should be considered when applying nucleic amplification techniques for characterising M. hyopneumoniae strains to specify the epidemiology of infection and to evaluate virulence factors triggering the corresponding disease.  相似文献   

16.
The applicability of conventional and molecular methods for rapid detection and differentiation of Pasteurella multocida serogroup B isolates involved in an outbreak of haemorrhagic septicaemia affecting Indian buffaloes, was studied. Five isolates were obtained and were subjected to phenotypic and genotypic characterization. None of the five isolates could be differentiated on the basis of cultural, biochemical, pathogenicity and antimicrobial sensitivity patterns. Polymerase chain reaction (PCR)-based techniques were found to be specific and sensitive for rapid detection and differentiation of isolates. Repetitive extragenic palindromic (REP-) PCR, enterobacterial repetitive intergenic consensus (ERIC-) PCR and single-primer PCR differentiated all the five isolates into different profiles. All the isolates involved in the outbreak were found to have a genetic profile different from standard P. multocida strain (P52). However, three isolates had similar profiles, whereas each of the remaining two had a different profile. The study indicates the involvement of multiple strains of P. multocida in a single outbreak of haemorrhagic septicaemia in buffaloes. The results also indicate that molecular methods of detection and typing are superior to conventional methods for rapid epidemiological investigations of haemorrhagic septicaemia.  相似文献   

17.
Twenty-nine isolates of Malassezia pachydermatis were recovered from a single farm of 100 pigs in Croatia. In contrast, 290 farm pigs from other locations (northern parts of Croatia and Slovenia) yielded only two non-lipid dependent isolates of M. pachydermatis using the same swabbing procedure. Ten of the 29 isolates from a single farm had their identity confirmed by karyotyping, and were typed by restriction fragment length polymorphism (RFLP) analysis. All but one of these isolates sub-typed were indistinguishable, one isolate produced a slightly different RFLP profile. Control isolates recovered from dog skin gave RFLP profiles that were easily distinguished from those produced by the pig isolates. These results suggest that a single strain of M. pachydermatis had colonised this pig herd.  相似文献   

18.
Molecular characterization of Blastocystis isolates from primates   总被引：2，自引：0，他引：2  
Twelve Blastocystis isolates from primates were analyzed genetically by polymerase chain reaction (PCR) using diagnostic primers and PCR-restriction fragment length polymorphism (RFLP) of SSUrDNA. Two distinct genotypes, subtype 1 and a variant of subtype 1, were detected in two and six of the 12 isolates, respectively. The RFLP profiles of the isolates designated as subtype 1 were identical to the profile of ribodeme 1. The RFLP profiles of the six isolates designated as variants of subtype 1 were different from the profile of the variant of subtype 1 from a human reported previously. The other four isolates were not amplified with any diagnostic primers, but three of them showed the same RFLP profiles as ribodeme 6. This study was the first genomic analysis of Blastocystis isolates from primates, and showed the genetic similarity between the isolates from primates and the genotypes of Blastocystis hominis. However, it was unclear whether the isolates examined were zoonotic or not. Therefore, it is necessary to reveal the phylogenetic relationships between the isolates from primates and the multiple genotypes of B. hominis.  相似文献   

19.
IntroductionCutaneous leishmaniasis (CL) is one of wobbling endemic disease in Iraq, that cause intracellular obligate protistan parasite returned to the genusLeishmania. This study is aimed to identify epidemiology of CL, detect the prevalence of Leishmania tropica and find the phylogenetic relationship.MethodologyThe current study was conducted in the main hospitals of Thi-Qar province-south of Iraq for period from November 2018 to October 2019. Nested-PCR was used to amplify kinetoplast minicircle fragments DNA.ResultsIt was recorded 247 clinical cases with CL, the infections of males were higher than females, while infection rate appeared gradual reduction with age progress. Furthermore, the most CL infections were as single lesions and occurred in December. The infections of upper limbs were high when compared with other body regions. The molecular diagnosis showed L. tropica was more frequently. DNA sequences of kDNA gene of L. tropica showed confirmative genetic detection of local isolates using NCBI-Blast data and phylogenetic tree analysis after comparison with global recorded isolates. The local L. tropica isolates showed genetically closed related to NCBI-Blast L. tropica with accession number AB678350.1. Generally, the analysis of kDNA nitrogen bases sequences showed that all of samples were consistent with those recorded at the NCBI.ConclusionThe kDNA minicircle sequences analysis results showed mismatching of the local isolates decrease whenever approached from the Iranian border. In addition, genetic heterogeneity diagnosis is important for detection of therapy, control and epidemiological studies.  相似文献   

20.
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