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1.
PLoS One ; 18(8): e0290473, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37616210

RESUMO

Understanding the microbial genomic contributors to antimicrobial resistance (AMR) is essential for early detection of emerging AMR infections, a pressing global health threat in human and veterinary medicine. Here we used whole genome sequencing and antibiotic susceptibility test data from 980 disease causing Escherichia coli isolated from companion and farm animals to model AMR genotypes and phenotypes for 24 antibiotics. We determined the strength of genotype-to-phenotype relationships for 197 AMR genes with elastic net logistic regression. Model predictors were designed to evaluate different potential modes of AMR genotype translation into resistance phenotypes. Our results show a model that considers the presence of individual AMR genes and total number of AMR genes present from a set of genes known to confer resistance was able to accurately predict isolate resistance on average (mean F1 score = 98.0%, SD = 2.3%, mean accuracy = 98.2%, SD = 2.7%). However, fitted models sometimes varied for antibiotics in the same class and for the same antibiotic across animal hosts, suggesting heterogeneity in the genetic determinants of AMR resistance. We conclude that an interpretable AMR prediction model can be used to accurately predict resistance phenotypes across multiple host species and reveal testable hypotheses about how the mechanism of resistance may vary across antibiotics within the same class and across animal hosts for the same antibiotic.


Assuntos
Antibacterianos , Gado , Animais , Humanos , Antibacterianos/farmacologia , Animais de Estimação , Farmacorresistência Bacteriana/genética , Escherichia coli/genética
2.
Front Vet Sci ; 8: 674307, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34414224

RESUMO

Mycobacterium bovis causes tuberculosis (TB) in cattle, which in turn can transmit the pathogen to humans. Tuberculosis in dairy cattle is of particular concern where the consumption of raw milk and dairy products is customary. Baja California (BCA), Mexico, presents high prevalence of TB in both cattle and humans, making it important to investigate the molecular epidemiology of the disease in the region. A long-term study was undertaken to fully characterize the diversity of M. bovis genotypes circulating in dairy cattle, cheese and humans in BCA by whole-genome sequencing (WGS). During a 2-year period, 412 granulomatous tissue samples were collected from local abattoirs and 314 cheese samples were purchased from local stores and vendors in BCA and sent to the laboratory for mycobacterial culture, histology, direct PCR and WGS. For tissue samples M. bovis was recovered from 86.8%, direct PCR detected 90% and histology confirmed 85.9% as mycobacteriosis-compatible. For cheese, M. bovis was recovered from 2.5% and direct PCR detected 6% of the samples. There was good agreement between diagnostic tests. Subsequently, a total of 345 whole-genome SNP sequences were obtained. Phylogenetic analysis grouped these isolates into 10 major clades. SNP analysis revealed putative transmission clusters where the pairwise SNP distance between isolates from different dairies was ≤3 SNP. Also, human and/or cheese isolates were within 8.45 (range 0-17) and 5.8 SNP (range 0-15), respectively, from cattle isolates. Finally, a comparison between the genotypes obtained in this study and those reported previously suggests that the genetic diversity of M. bovis in BCA is well-characterized, and can be used to determine if BCA is the likely source of M. bovis in humans and cattle in routine epidemiologic investigations and future studies. In conclusion, WGS provided evidence of ongoing local transmission of M. bovis among the dairies in this high-TB burden region of BCA, as well as show close relationships between isolates recovered from humans, cheese, and cattle. This confirms the need for a coordinated One Health approach in addressing the elimination of TB in animals and humans. Overall, the study contributes to the knowledge of the molecular epidemiology of M. bovis in BCA, providing insight into the pathogen's dynamics in a high prevalence setting.

3.
PLoS One ; 16(5): e0246983, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33983941

RESUMO

Recent evidence of circulation of multiple strains within herds and mixed infections of cows marks the beginning of a rethink of our knowledge on Mycobacterium avium ssp. paratuberculosis (MAP) epidemiology. Strain typing opens new ways to investigate MAP transmission. This work presents a method for reconstructing infection chains in a setting of endemic Johne's disease on a well-managed dairy farm. By linking genomic data with demographic field data, strain-specific differences in spreading patterns could be quantified for a densely sampled dairy herd. Mixed infections of dairy cows with MAP are common, and some strains spread more successfully. Infected cows remain susceptible for co-infections with other MAP genotypes. The model suggested that cows acquired infection from 1-4 other cows and spread infection to 0-17 individuals. Reconstructed infection chains supported the hypothesis that high shedding animals that started to shed at an early age and showed a progressive infection pattern represented a greater risk for spreading MAP. Transmission of more than one genotype between animals was recorded. In this farm with a good MAP control management program, adult-to-adult contact was proposed as the most important transmission route to explain the reconstructed networks. For each isolate, at least one more likely ancestor could be inferred. Our study results help to capture underlying transmission processes and to understand the challenges of tracing MAP spread within a herd. Only the combination of precise longitudinal field data and bacterial strain type information made it possible to trace infection in such detail.


Assuntos
Doenças dos Bovinos/microbiologia , Cadeia de Infecção/veterinária , Indústria de Laticínios , Doenças Endêmicas/veterinária , Genômica , Mycobacterium avium subsp. paratuberculosis/fisiologia , Paratuberculose/genética , Paratuberculose/microbiologia , Animais , Bovinos , Doenças dos Bovinos/genética , Feminino , Funções Verossimilhança , Fenótipo , Filogenia
4.
Mol Ecol ; 28(9): 2192-2205, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30807679

RESUMO

The role of wildlife in the persistence and spread of livestock diseases is difficult to quantify and control. These difficulties are exacerbated when several wildlife species are potentially involved. Bovine tuberculosis (bTB), caused by Mycobacterium bovis, has experienced an ecological shift in Michigan, with spillover from cattle leading to an endemically infected white-tailed deer (deer) population. It has potentially substantial implications for the health and well-being of both wildlife and livestock and incurs a significant economic cost to industry and government. Deer are known to act as a reservoir of infection, with evidence of M. bovis transmission to sympatric elk and cattle populations. However, the role of elk in the circulation of M. bovis is uncertain; they are few in number, but range further than deer, so may enable long distance spread. Combining Whole Genome Sequences (WGS) for M. bovis isolates from exceptionally well-observed populations of elk, deer and cattle with spatiotemporal locations, we use spatial and Bayesian phylogenetic analyses to show strong spatiotemporal admixture of M. bovis isolates. Clustering of bTB in elk and cattle suggests either intraspecies transmission within the two populations, or exposure to a common source. However, there is no support for significant pathogen transfer amongst elk and cattle, and our data are in accordance with existing evidence that interspecies transmission in Michigan is likely only maintained by deer. This study demonstrates the value of whole genome population studies of M. bovis transmission at the wildlife-livestock interface, providing insights into bTB management in an endemic system.


Assuntos
Cervos/microbiologia , Mycobacterium bovis/genética , Tuberculose Bovina/transmissão , Tuberculose/veterinária , Animais , Bovinos , Interações Hospedeiro-Patógeno , Gado/microbiologia , Michigan , Mycobacterium bovis/isolamento & purificação , Mycobacterium bovis/patogenicidade , Filogenia , Análise Espaço-Temporal , Tuberculose/transmissão , Tuberculose Bovina/prevenção & controle , Sequenciamento Completo do Genoma
5.
Int J Infect Dis ; 63: 48-56, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28739421

RESUMO

OBJECTIVES: To determine genetic diversity by comparing the whole genome sequences of cattle and human Mycobacterium bovis isolates from Baja California. METHODS: A whole genome sequencing strategy was used to obtain the molecular fingerprints of 172 isolates of M. bovis obtained from Baja California, Mexico; 155 isolates were from cattle and 17 isolates were from humans. Spoligotypes were characterized in silico and single nucleotide polymorphism (SNP) differences between the isolates were evaluated. RESULTS: A total of 12 M. bovis spoligotype patterns were identified in cattle and humans. Two predominant spoligotypes patterns were seen in both cattle and humans: SB0145 and SB1040. The SB0145 spoligotype represented 59% of cattle isolates (n=91) and 65% of human isolates (n=11), while the SB1040 spoligotype represented 30% of cattle isolates (n=47) and 30% of human isolates (n=5). When evaluating SNP differences, the human isolates were intimately intertwined with the cattle isolates. CONCLUSIONS: All isolates from humans had spoligotype patterns that matched those observed in the cattle isolates, and all human isolates shared common ancestors with cattle in Baja California based on SNP analysis. This suggests that most human tuberculosis caused by M. bovis in Baja California is derived from M. bovis circulating in Baja California cattle. These results reinforce the importance of bovine tuberculosis surveillance and control in this region.


Assuntos
Mycobacterium bovis/genética , Mycobacterium bovis/isolamento & purificação , Tuberculose Bovina/epidemiologia , Tuberculose/epidemiologia , Sequenciamento Completo do Genoma , Animais , Técnicas de Tipagem Bacteriana , Bovinos , DNA Bacteriano/genética , Humanos , México/epidemiologia , Polimorfismo de Nucleotídeo Único
6.
J Am Vet Med Assoc ; 251(2): 206-216, 2017 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-28671497

RESUMO

OBJECTIVE To describe use of whole-genome sequencing (WGS) and evaluate the apparent sensitivity and specificity of antemortem tuberculosis tests during investigation of an unusual outbreak of Mycobacterium bovis infection in a Michigan dairy herd. DESIGN Bovine tuberculosis (bTB) outbreak investigation. ANIMALS Cattle, cats, dog, and wildlife. PROCEDURES All cattle in the index dairy herd were screened for bTB with the caudal fold test (CFT), and cattle ≥ 6 months old were also screened with a γ-interferon (γIFN) assay. The index herd was depopulated along with all barn cats and a dog that were fed unpasteurized milk from the herd. Select isolates from M bovis-infected animals from the index herd and other bTB-affected herds underwent WGS. Wildlife around all affected premises was examined for bTB. RESULTS No evidence of bTB was found in any wildlife examined. Within the index herd, 53 of 451 (11.8%) cattle and 12 of 21 (57%) cats were confirmed to be infected with M bovis. Prevalence of M bovis-infected cattle was greatest among 4- to 7-month-old calves (16/49 [33%]) followed by adult cows (36/203 [18%]). The apparent sensitivity and specificity were 86.8% and 92.7% for the CFT and 80.4% and 96.5% for the γIFN assay when results for those tests were interpreted separately and 96.1% and 91.7% when results were interpreted in parallel. Results of WGS revealed that M bovis-infected barn cats and cattle from the index herd and 6 beef operations were infected with the same strain of M bovis. Of the 6 bTB-affected beef operations identified during the investigation, 3 were linked to the index herd only by WGS results; there was no record of movement of livestock or waste milk from the index herd to those operations. CONCLUSIONS AND CLINICAL RELEVANCE Whole-genome sequencing enhanced the epidemiological investigation and should be used in all disease investigations. Performing the CFT and γIFN assay in parallel improved the antemortem ability to detect M bovis-infected animals. Contact with M bovis-infected cattle and contaminated milk were major risk factors for transmission of bTB within and between herds of this outbreak.


Assuntos
Mycobacterium bovis/genética , Tuberculose Bovina/epidemiologia , Animais , Animais Recém-Nascidos , Bovinos , Surtos de Doenças/veterinária , Feminino , Michigan/epidemiologia , Tuberculose Bovina/microbiologia
7.
Genome Announc ; 5(14)2017 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-28385856

RESUMO

We report here an update to the reference genome sequence of the bovine tuberculosis bacillus Mycobacterium bovis AF2122/97, generated using an integrative multiomics approach. The update includes 42 new coding sequences (CDSs), 14 modified annotations, 26 single-nucleotide polymorphism (SNP) corrections, and disclosure that the RD900 locus, previously described as absent from the genome, is in fact present.

8.
Artigo em Inglês | MEDLINE | ID: mdl-27734009

RESUMO

Brucella are highly infectious bacterial pathogens responsible for brucellosis, a frequent worldwide zoonosis. The Brucella genus has recently expanded from 6 to 11 species, all of which were associated with mammals; The natural host range recently expanded to amphibians after some reports of atypical strains from frogs. Here we describe the first in depth phenotypic and genetic characterization of a Brucella strains isolated from a frog. Strain B13-0095 was isolated from a Pac-Man frog (Ceratophyrus ornate) at a veterinary hospital in Texas and was initially misidentified as Ochrobactrum anthropi. We found that B13-0095 belongs to a group of early-diverging brucellae that includes Brucella inopinata strain BO1 and the B. inopinata-like strain BO2, with traits that depart significantly from those of the "classical" Brucella spp. Analysis of B13-0095 genome sequence revealed several specific features that suggest that this isolate represents an intermediate between a soil associated ancestor and the host adapted "classical" species. Like strain BO2, B13-0095 does not possess the genes required to produce the perosamine based LPS found in classical Brucella, but has a set of genes that could encode a rhamnose based O-antigen. Despite this, B13-0095 has a very fast intracellular replication rate in both epithelial cells and macrophages. Finally, another major finding in this study is the bacterial motility observed for strains B13-0095, BO1, and BO2, which is remarkable for this bacterial genus. This study thus highlights several novel characteristics in strains belonging to an emerging group within the Brucella genus. Accurate identification tools for such atypical Brucella isolates and careful evaluation of their zoonotic potential, are urgently required.


Assuntos
Anuros/microbiologia , Brucella/classificação , Brucella/isolamento & purificação , Brucelose/microbiologia , Filogenia , Animais , Proteínas de Bactérias/genética , Sequência de Bases , Evolução Biológica , Brucella/genética , Brucella/metabolismo , Brucelose/mortalidade , Carbono/metabolismo , Linhagem Celular/patologia , Criança , DNA Bacteriano/genética , Células Epiteliais/microbiologia , Feminino , Genes Bacterianos , Genoma Bacteriano , Células HeLa/patologia , Humanos , Lipopolissacarídeos/classificação , Lipopolissacarídeos/genética , Macrófagos/microbiologia , Camundongos , Família Multigênica , Antígenos O/genética , Fenótipo , Ramnose/metabolismo , Texas , Virulência , Zoonoses/microbiologia
9.
Genome Announc ; 4(4)2016 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-27469947

RESUMO

Mycobacterium mungi, a Mycobacterium tuberculosis complex pathogen, has emerged in banded mongoose in northern Botswana and Northwest Zimbabwe. The pathogen is transmitted through infected secretions used in olfactory communication behavior (K. A. Alexander, C. E. Sanderson, M. H. Larsen, S. Robbe-Austerman, M. C. Williams, and M. V. Palmer, mBio 7(3):e00281-16, 2016, http://dx.doi.org/10.1128/mBio.00281-16). We announce here the draft genome sequence of this emerging pathogen.

10.
Vet Microbiol ; 180(1-2): 75-81, 2015 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-26243696

RESUMO

Mycobacterium bovis (M. bovis) causes tuberculosis in white-tailed deer (WTD). Natural infection of WTD with M. bovis is most closely mimicked by instilling inoculum into palatine tonsillar crypts. One hundred fifty days after intratonsillar inoculation, M. bovis was cultured from 30 tissues originating from 14 deer. Whole-genome sequencing (WGS) was performed on the original inoculum, single colonies subcultured from the original inoculum, and M. bovis isolated from each culture positive tissue. Single nucleotide polymorphisms (SNP) were identified by comparing the derived sequences to the reference strain AF2122/97. Results indicate that the majority of the SNPs that were identified were homogeneous between the inoculum and the isolates from the tissues. The majority of individual tissues had different WGS genotypes from each other, suggesting that dissemination of M. bovis beyond the initial site of infection may require few mycobacteria representing a bottleneck.


Assuntos
Cervos/microbiologia , Genótipo , Mycobacterium bovis/genética , Mycobacterium bovis/isolamento & purificação , Tuberculose/veterinária , Animais , Mycobacterium bovis/classificação , Tonsila Palatina/microbiologia , Polimorfismo de Nucleotídeo Único , Tuberculose/microbiologia
11.
Vaccine ; 31(44): 5082-7, 2013 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-24016805

RESUMO

Clostridium sordellii is a Gram positive anaerobic bacterium that causes multiple disease syndromes in both humans and animals. As with many clostridial pathogens, toxins contribute to the virulence of C. sordellii. Two large toxins have been identified: a lethal toxin (TcsL) and a hemorrhagic toxin (TcsH) which are similar in structure and function to Clostridium difficile toxin B (TcdB) and toxin A (TcdA), respectively. While TcdA, TcdB, and TcsL have been extensively studied, relatively little is known about TcsH. This study elucidated the TcsH gene sequence using whole genome sequencing, compared the genotype with toxin expression of 52 C. sordellii strains, and examined the role of TcsH in batch release potency tests required for veterinary vaccines licensed in the United States and other testing utilizing WHO standard antitoxin. Data from this study will assist in future research to clarify the TcsH contribution to the pathogenesis of C. sordellii infections and may aid in the development of improved vaccines.


Assuntos
Toxinas Bacterianas/genética , Clostridium sordellii/genética , Vacinação/veterinária , Animais , Anticorpos Monoclonais/imunologia , Antitoxinas/imunologia , Toxinas Bacterianas/imunologia , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/normas , Clostridium sordellii/patogenicidade , DNA Bacteriano/genética , Enterotoxinas/imunologia , Ensaio de Imunoadsorção Enzimática , Genes Bacterianos , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Testes de Toxicidade , Estados Unidos , Vacinação/normas , Virulência
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