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1.
Animals (Basel) ; 13(9)2023 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-37174606

RESUMO

(1) Background: paratuberculosis is an important disease in ruminants, causing worldwide economic losses to the livestock industry. Although vaccination is known not to prevent transmission of the causative agent Mycobacterium avium subsp. paratuberculosis (Map), it is considered an effective tool for paratuberculosis in infected herds. The objectives of this controlled field study were to evaluate the effects of the whole-cell heat-killed Silirum® vaccine on Map fecal shedding and serological status in dairy herds infected with paratuberculosis. (2) Methods: The serological status (ELISA) and fecal shedding (qPCR) of 358 vaccinated cows were assessed over 3 years in 7 infected dairy herds in the Meuse department, France. Within each herd, cows from the last non-vaccinated birth cohort (n = 265) were used as controls. The probability and level of Map fecal shedding and the serological status were modeled using multivariable mixed general linear regression models. (3) Results: Overall, 34.7% of cows tested positive at least once on fecal qPCR, with significant differences between herds, but high shedding levels were observed in only 5.5% of cows. Compared to non-vaccinated seronegative cows, a statistically significant reduction in the probability of Map shedding was found only in cows vaccinated before 4 to 5 months of age that tested negative for Map antibodies throughout the study period (odds ratio = 0.5, 95% confidence interval: 0.3-0.9, p = 0.008), but no significant effect of vaccination on the amount of Map shedding could be evidenced. Finally, the younger the cows were when vaccinated, the less they tested positive on the serum ELISA. (4) Conclusions: a beneficial effect of vaccination on Map fecal shedding may exist in cows vaccinated before 4 to 5 months of age. The variability of the serum ELISA response in vaccinated cows remains to be investigated.

2.
Front Immunol ; 13: 871780, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35677047

RESUMO

Recent developments in multiplex technologies enable the determination of a large nu\mber of soluble proteins such as cytokines in various biological samples. More than a one-by-one determination of the concentration of immune mediators, they permit the establishment of secretion profiles for a more accurate description of conditions related to infectious diseases or vaccination. Cytokine profiling has recently been made available for bovine species with the development of a Luminex® technology-based 15-plex assay. Independently from the manufacturer, we evaluated the bovine cytokine/chemokine multiplex assay for limits of detection, recovery rate, and reproducibility. Furthermore, we assessed cytokine secretion in blood samples from 107 cows upon stimulation with heat-killed bacteria and TLR2/4 ligands compared to a null condition. Secretion patterns were analyzed either using the absolute concentration of cytokines or using their relative concentration with respect to the overall secretion level induced by each stimulus. Using Partial Least Square-Discriminant Analysis, we show that the 15-cytokine profile is different under Escherichia coli, Staphylococcus aureus, and Streptococcus uberis conditions, and that IFN-γ, IL-1ß, and TNF-α contribute the most to differentiate these conditions. LPS and E. coli induced largely overlapping biological responses, but S. aureus and S. uberis were associated with distinct cytokine profiles than their respective TLR ligands. Finally, results based on adjusted or absolute cytokine levels yielded similar discriminative power, but led to different stimuli-related signatures.


Assuntos
Bovinos , Citocinas , Receptores Toll-Like , Animais , Bovinos/sangue , Citocinas/sangue , Escherichia coli , Feminino , Ligantes , Reprodutibilidade dos Testes , Staphylococcus aureus , Streptococcus , Receptores Toll-Like/imunologia
3.
Vet Rec ; 191(2): e1733, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35700231

RESUMO

BACKGROUND: The condemnation of carcases and offal unfit for human consumption is a regulatory requirement at the slaughterhouse. Condemnation data, if comprehensive and standardised, can be a valuable source of information for risk-based inspection and decision making. METHODS: The aim of this study was to analyse postmortem condemnation data that were recorded in all bovine slaughterhouses in mainland France from 1 January 2016 to 31 December 2020 in a comprehensive and standardised information system. The rates of and reasons for condemnation, as well as factors influencing rate variation, were investigated through descriptive analysis and multivariable logistic regression models. RESULTS: The global, total and partial condemnation rates were 4.5%, 0.7% and 3.8% for adult cattle and 1.4%, 0.3% and 1.1% for calves, respectively. Reasons for condemnation varied with the animal category; for example, the three main reasons for total condemnation in adult cattle were serous infiltration of connective tissue (49% of condemned animals), congestive peritonitis (12.2%) and fibrinous peritonitis (10.9%), whereas the top three reasons for partial condemnation were unique abscess (21.9%), haemorrhagic infiltration (20.6%) and muscular sclerosis (17.4%). Condemnation rates were influenced by animal-related factors (sex, age, type of breed) and slaughterhouse-related factors (status, type, slaughter volume). CONCLUSION: Our findings could usefully contribute to the continuous improvement of the harmonisation of inspection decisions and support the risk manager's strategy in the modernisation of official controls at the slaughterhouse.


Assuntos
Matadouros/estatística & dados numéricos , Doenças dos Bovinos/epidemiologia , Carne , Peritonite/veterinária , Matadouros/legislação & jurisprudência , Animais , Bovinos , França/epidemiologia , Humanos , Sistemas de Informação , Modelos Logísticos , Carne/normas , Análise Multivariada , Peritonite/classificação , Peritonite/epidemiologia
4.
Parasit Vectors ; 13(1): 529, 2020 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-33092627

RESUMO

BACKGROUND: Bovine besnoitiosis, an emerging disease in Europe that can be transmitted by vectors, is caused by the apicomplexan Besnoitia besnoiti. Bovine besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine besnoitiosis. METHODS: Previous real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (n = 518) from eight dairy or beef cattle farms facing a high serological prevalence of besnoitiosis were sampled at the tail base and their skin sample analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time. RESULTS: Among the 518 seropositive animals, a low proportion of individuals (14.5%) showed Cq values below 36, 17.8% had doubtful results (36 < Cq ≤ 40) and 67.8% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographical location or history of besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. CONCLUSIONS: Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine besnoitiosis control.


Assuntos
Portador Sadio/veterinária , Bovinos/parasitologia , Coccidiose/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sarcocystidae/isolamento & purificação , Animais , Anticorpos Antiprotozoários/sangue , Biópsia , Portador Sadio/parasitologia , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/prevenção & controle , Coccidiose/diagnóstico , Coccidiose/prevenção & controle , DNA de Protozoário , Testes Diagnósticos de Rotina/métodos , Patologia Molecular , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sarcocystidae/genética , Sarcocystidae/imunologia , Testes Sorológicos , Pele/parasitologia
5.
PLoS One ; 14(12): e0226246, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31877160

RESUMO

The aim of our study was to evaluate the flock sensitivity and specificity of fecal qPCR and serum ELISA using pooled samples for screening paratuberculosis in French sheep. Using individual feces with low or high qPCR Ct values from ewes sampled in 14 infected flocks, a total of 555 pools of size 5, 10 and 20 were created by diluting individual materials in negative feces and analysed using a commercial IS900 qPCR kit. The relative performances of pooled serum ELISA analysis were evaluated based on the analysis of 181 different pools of size 5 and 10, composed of individual serum samples of various individual S/P values. Results showed that for pools of size 5, 10 or 20, individual fecal samples with low Ct values were invariably detected. Conversely fecal samples with high Ct values were associated with a lower detection rate in both pools of size 5 (87.0% to 90.0%), 10 (63.0% to 70.7%) and 20 (46.7% to 60.0%). After lowering the decision threshold to 25% and 15% for serum pools of size 5 and 10 respectively, the pooled serum ELISA relative sensitivity ranged between 62.2% and 100.0% depending on the composition of the pools. Finally, a simulation study was carried out to evaluate the performances of 16 screening strategies at flock level, with varying pool size (5 to 20) and number (5 to 60). The use of pooled serum ELISA led to very false positive detection rate ranging between 37.6% and 91.8% in paratuberculosis free flocks and prevents its further use in that context. For infection prevalence ≤ 5%, the flock sensitivity based on pooled fecal qPCR ranged between 39.0% (5 pools of size 10) and 99.9% (300 sampled individuals, with pools of size 5,10 or20), and was always above 93% when the infection prevalence was greater or equal to 15%. We conclude that pooled-fecal qPCR but not pooled-serum ELISA could be a useful tool to detect sheep flocks infected with paratuberculosis.


Assuntos
Anticorpos Antibacterianos/sangue , Fezes/microbiologia , Mycobacterium avium subsp. paratuberculosis/genética , Paratuberculose/diagnóstico , Doenças dos Ovinos/microbiologia , Animais , Estudos Transversais , Diagnóstico Precoce , Ensaio de Imunoadsorção Enzimática/veterinária , Mycobacterium avium subsp. paratuberculosis/imunologia , Paratuberculose/diagnóstico por imagem , Prevalência , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sensibilidade e Especificidade , Ovinos , Doenças dos Ovinos/epidemiologia , Manejo de Espécimes
6.
Front Vet Sci ; 6: 66, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30895182

RESUMO

Risk factors are key epidemiological concepts that are used to explain disease distributions. Identifying disease risk factors is generally done by comparing the characteristics of diseased and non-diseased populations. However, imperfect disease detectability generates disease observations that do not necessarily represent accurately the true disease situation. In this study, we conducted an extensive simulation exercise to emphasize the impact of imperfect disease detection on the outcomes of logistic models when case reports are aggregated at a larger scale (e.g., diseased animals aggregated at farm level). We used a probabilistic framework to simulate both the disease distribution in herds and imperfect detectability of the infected animals in these herds. These simulations show that, under logistic models, true herd-level risk factors are generally correctly identified but their associated odds ratio are heavily underestimated as soon as the sensitivity of the detection is less than one. If the detectability of infected animals is not only imperfect but also heterogeneous between herds, the variables associated with the detection heterogeneity are likely to be incorrectly identified as risk factors. This probability of type I error increases with increasing heterogeneity of the detectability, and with decreasing sensitivity. Finally, the simulations highlighted that, when count data is available (e.g., number of infected animals in herds), they should not be reduced to a presence/absence dataset at the herd level (e.g., presence or not of at least one infected animal) but rather modeled directly using zero-inflated count models which are shown to be much less sensitive to imperfect detectability issues. In light of these simulations, we revisited the analysis of the French bovine abortion surveillance data, which has already been shown to be characterized by imperfect and heterogeneous abortion detectability. As expected, we found substantial differences between the quantitative outputs of the logistic model and those of the zero-inflated Poisson model. We conclude by strongly recommending that efforts should be made to account for, or at the very least discuss, imperfect disease detectability when assessing associations between putative risk factors and observed disease distributions, and advocate the use of zero-inflated count models if count data is available.

7.
J Contemp Brachytherapy ; 10(5): 463-469, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30479624

RESUMO

PURPOSE: The aim of this study is to investigate the feasibility of perioperative I-125 low-dose-rate brachytherapy mesh implantation in pelvic locations in an animal model, before applying it clinically. MATERIAL AND METHODS: The animal model was the Romanov adult ewe. Non-radioactive dummy I-125 seeds were implanted by laparotomy in the pelvic area. Forty-five dummy seeds were placed on a 10 cm² polyglactin mesh to obtain a dose of 160 Gy at 5 mm from the center of each seed. Three CT scans were performed at day 15, day 70, and day 180 after surgery to check the positioning of the mesh for eventual seed migration according to bony landmarks and to perform a 3D theoretical dosimetric study. The experimental study design was based on Simon's minimax plan with a preliminary analysis of 10 ewes to validate the protocol and a second series of 7 ewes. RESULTS: After the first step, 9 of 10 ewes were investigated. For 8 of 9 animals, the 160 Gy isodose line volume was within 10%, showing feasibility of the procedure and allowing 7 more to be added. At the end of the study, 16 of 17 animals were examined. No seeds loss was observed. The volume difference of the 160 Gy isodose line was within 10% in 13 of 16 ewes between the three CT scans. Twelve out of 16 had a coordinate deviation less than or equal to 10 mm on the three axes between the first and the third scans. CONCLUSIONS: These results show the technical feasibility of the pelvic mesh implantation in ewes. A phase I study for patients with locally advanced or recurrent pelvic tumors amenable to surgery, in combination with surgical resection should be possible.

8.
PLoS One ; 13(7): e0199914, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29975760

RESUMO

Bovine spongiform encephalopathy (BSE) can be efficiently transmitted to pigs via intracerebral inoculation. A clear link has been established between the consumption of products of bovine origin contaminated with the BSE agent and the development of variant Creutzfeldt-Jakob disease in humans. Small ruminants can also naturally develop BSE, and sheep-adapted BSE (Sh-BSE) propagates more efficiently than cattle BSE in pigs and in mouse models expressing porcine prion protein. In addition, Sh-BSE shows greater efficiency of transmission to human models than original cow BSE. While infectivity and/or abnormal PrP accumulation have been reported in the central nervous system in BSE-infected pigs, the ability of the agent to replicate in peripheral tissues has not been fully investigated. We previously characterized the presence of prions in a panel of tissues collected at the clinical stage of disease from pigs experimentally infected with Sh-BSE. Western blot revealed low levels of PrPres accumulation in lymphoid tissues, nerves, and skeletal muscles from 4 of the 5 animals analysed. Using protein misfolding cyclic amplification (PMCA), which we found to be 6 log fold more sensitive than direct WB for the detection of pig BSE, we confirmed the presence of the Sh-BSE agent in lymphoid organs, nerves, ileum, and striated muscles from all 5 inoculated pigs. Surprisingly, PrPres positivity was also detected in white blood cells from one pig using this method. The presence of infectivity in lymphoid tissues, striated muscles, and peripheral nerves was confirmed by bioassay in bovine PrP transgenic mice. These results demonstrate the ability of BSE-derived agents to replicate efficiently in various peripheral tissues in pigs. Although no prion transmission has been reported in pigs following oral BSE challenge, our data support the continuation of the Feed Ban measure implemented to prevent entry of the BSE agent into the feed chain.


Assuntos
Encéfalo/metabolismo , Encefalopatia Espongiforme Bovina/transmissão , Nervos Periféricos/metabolismo , Proteínas PrPSc/metabolismo , Doenças dos Ovinos/transmissão , Animais , Encéfalo/patologia , Bovinos , Modelos Animais de Doenças , Encefalopatia Espongiforme Bovina/metabolismo , Encefalopatia Espongiforme Bovina/patologia , Camundongos , Camundongos Transgênicos , Nervos Periféricos/patologia , Proteínas PrPSc/isolamento & purificação , Ovinos , Doenças dos Ovinos/metabolismo , Doenças dos Ovinos/patologia
9.
J Gen Virol ; 98(10): 2628-2634, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28920852

RESUMO

Multiple theories exist regarding the origin of bovine spongiform encephalopathy (BSE). An early and prominent theory proposed that BSE was the result of the adaptation of sheep scrapie to cattle. The reports to date indicate that the distribution of the pathological prion protein (PrPSc) in experimental bovine scrapie is largely restricted to the central nervous system (CNS). Here, we describe pathological findings in a calf intracerebrally inoculated with a Spanish classical scrapie isolate. While clinical disease was observed 30 months after inoculation and PrPSc was detected in the CNS, the corresponding phenotype differed from that of BSE. Immunohistochemistry and PMCA also revealed the presence of PrPSc in the peripheral nerves, lymphoid tissues, skeletal muscle and gastrointestinal tract, suggesting centrifugal spread of the scrapie agent from the brain. To the best of our knowledge, this is the first report describing the detection of PrPSc in tissues other than the CNS after experimental transmission of scrapie to cattle.

10.
BMC Vet Res ; 13(1): 230, 2017 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-28774299

RESUMO

BACKGROUND: The objective was to evaluate the diagnostic accuracy of two serum ELISAs and one quantitative PCR on feces for the diagnosis of paratuberculosis in sub-clinically infected young-adult sheep. A cross-sectional study was performed to collect 1197 individual blood and fecal samples from 2- to 3-year-old sub-clinically infected ewes in 14 closed meat sheep flocks in France. Fecal excretion was determined using qPCR based on IS900 sequence detection, and serology was performed on serum samples using two commercial ELISAs. Data were analyzed in a 3-test multiple-population Bayesian latent class model accounting for potential dependence between the three tests fitted in OpenBUGS. Separate analyses were performed according to whether doubtful ELISA results were handled as positive or negative and based on two thresholds for fecal qPCR (Ct ≤ 42 or Ct ≤ 40). RESULTS: The best fit to the data was provided by accounting for a pairwise dependence between the two ELISAs on sensitivity and pairwise dependence between the three tests on specificity. Under this model, the estimated ELISA sensitivities were 17.4% (95% PCI: 10.6 - 25.9) and 17.9% (95% PCI 11.4 - 25.6), with estimated specificities of 94.8% (95% PCI: 93.1 - 96.3) and 94.0% (95% PCI: 92.2 - 95.7). Fecal qPCR demonstrated significantly higher sensitivity (47.5%; 95% PCI: 29.3 - 69.9) and specificity (99.0%; 95% PCI: 97.9 - 99.9) than the ELISAs. Assumptions regarding doubtful ELISA results and qPCR thresholds had only a slight impact on test accuracy estimates. Models not accounting for pairwise dependence between ELISA and fecal qPCR results yielded higher sensitivity and specificity estimates but always provided a worse fit to the data. CONCLUSIONS: Although the overall sensitivity of serum ELISAs and fecal qPCR remains low, the higher diagnostic performances of fecal qPCR make it more suitable for paratuberculosis diagnosis in sub-clinically infected sheep. Our results also illustrate that all dependence structures should be investigated when evaluating diagnostic test accuracy and selection based on a rigorous statistical approach.


Assuntos
Ensaio de Imunoadsorção Enzimática/veterinária , Mycobacterium avium subsp. paratuberculosis , Paratuberculose/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Doenças dos Ovinos/diagnóstico , Animais , Teorema de Bayes , Estudos Transversais , Ensaio de Imunoadsorção Enzimática/métodos , Fezes/microbiologia , Feminino , França , Mycobacterium avium subsp. paratuberculosis/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade , Ovinos
11.
J Gen Virol ; 98(8): 2200-2204, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28721847

RESUMO

The ARR allele is considered to provide a very strong resistance against classical scrapie infection in sheep. In this study, we report the occurrence of clinical transmissible spongiform encephalopathy in ARR/ARR sheep, following their inoculation by the intracerebral route with a classical scrapie isolate. On first passage, the disease displayed an incomplete attack rate transmission, with incubation periods exceeding 6 years. On second passage, the obtained prion did not display better abilities to propagate than the original isolate. These transmission results contrasted with the 100 % attack rate and the short incubation periods observed in ARQ/ARQ sheep challenged with the same isolate. These data confirm that ARR/ARR sheep cannot be considered to be fully resistant to classical scrapie. However, they also support the contention that classical scrapie has a very limited capacity to transmit and adapt to ARR/ARR sheep.


Assuntos
Príons/genética , Scrapie/genética , Doenças dos Ovinos/genética , Ovinos/genética , Animais , Genótipo , Camundongos , Príons/metabolismo , Scrapie/metabolismo , Scrapie/transmissão , Ovinos/metabolismo , Doenças dos Ovinos/metabolismo , Doenças dos Ovinos/transmissão
12.
J Virol ; 90(7): 3439-45, 2016 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-26764000

RESUMO

UNLABELLED: Previous experiments carried out in a sheep scrapie model demonstrated that the transfusion of 200 µl of prion-infected whole blood has an apparent 100% efficacy for disease transmission. These experiments also indicated that, despite the apparent low infectious titer, the intravenous administration of white blood cells (WBC) resulted in efficient disease transmission. In the study presented here, using the same transmissible spongiform encephalopathy (TSE) animal model, our aim was to determine the minimal number of white blood cells and the specific abilities of mononucleated cell populations to transmit scrapie by the transfusion route. Our results confirmed that the transfusion of 100 µl, but not 10 µl, of fresh whole blood collected in asymptomatic scrapie-infected donor sheep can transmit the disease. The data also show that the intravenous administration of 10(5) WBCs is sufficient to cause scrapie in recipient sheep. Cell-sorted CD45R(+) (predominantly B lymphocytes), CD4(+)/CD8(+) (T lymphocytes), and CD14(+) (monocytes/macrophages) blood cell subpopulations all were shown to contain prion infectivity by bioassays in ovine PrP transgenic mice. However, while the intravenous administration of 10(6) CD45(+) or CD4(+)/8(+) living cells was able to transmit the disease, similar numbers of CD14(+) cells failed to infect the recipients. These data support the contention that mononucleated blood cell populations display different abilities to transmit TSE by the transfusion route. They also represent an important input for the risk assessment of blood-borne prion disease transmission and for refining the target performance of leukoreduction processes that currently are applied to mitigate the transmission risk in transfusion medicine. IMPORTANCE: Interindividual variant Creutzfeldt-Jakob disease (vCJD) transmission through blood and blood-derived products is considered a major public health issue in transfusion medicine. Over the last decade, TSE in sheep has emerged as a relevant model for assessing the blood-borne vCJD transmission risk. In this study, using a sheep TSE model, we characterized the ability of different peripheral blood mononucleated cell populations to infect TSE-free recipients by the transfusion route. Our results indicate that as little as 10(5) WBC and 100 µl of blood collected from asymptomatic scrapie infected animals can transmit the disease. They also demonstrate unambiguously that peripheral blood mononuclear cell subpopulations display dramatically different abilities to transmit the disease. These data represent an important input for the risk assessment of blood-borne prion disease transmission and for refining the target performance of leukoreduction processes that currently are applied to mitigate the transmission risk in transfusion medicine.


Assuntos
Leucócitos Mononucleares/transplante , Scrapie/sangue , Scrapie/transmissão , Reação Transfusional , Animais , Linfócitos B/transplante , Síndrome de Creutzfeldt-Jakob/sangue , Síndrome de Creutzfeldt-Jakob/transmissão , Modelos Animais de Doenças , Macrófagos/transplante , Camundongos , Ovinos , Linfócitos T/transplante
13.
Parasit Vectors ; 7: 439, 2014 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-25228371

RESUMO

BACKGROUND: Anaplasma phagocytophilum is a tick-borne intragranulocytic alpha-proteobacterium. It is the causative agent of tick-borne fever in ruminants, and of human granulocytic anaplasmosis in humans, two diseases which are becoming increasingly recognized in Europe and the USA. However, while several molecular typing tools have been developed over the last years, few of them are appropriate for in-depth exploration of the epidemiological cycle of this bacterium. Therefore we have developed a Multiple-Locus Variable number tandem repeat (VNTR) Analysis typing technique for A. phagocytophilum. METHODS: Five VNTRs were selected based on the HZ human-derived strain genome, and were tested on the Webster human-derived strain and on 123 DNA samples: 67 from cattle, 7 from sheep, 15 from roe deer, 4 from red deer, 1 from a reindeer, 2 from horses, 1 from a dog, and 26 from ticks. RESULTS: From these samples, we obtained 84 different profiles, with a diversity index of 0.96 (0.99 for vertebrate samples, i.e. without tick samples). Our technique confirmed that A. phagocytophilum from roe deer or domestic ruminants belong to two different clusters, while A. phagocytophilum from red deer and domestic ruminants locate within the same cluster, questioning the respective roles of roe vs red deer as reservoir hosts for domestic ruminant strains in Europe. As expected, greater diversity was obtained between rather than within cattle herds. CONCLUSIONS: Our technique has great potential to provide detailed information on A. phagocytophilum isolates, improving both epidemiological and phylogenic investigations, thereby helping in the development of relevant prevention and control measures.


Assuntos
Anaplasma phagocytophilum/genética , Ehrlichiose/veterinária , Repetições Minissatélites/genética , Ruminantes , Sequências de Repetição em Tandem/genética , Doenças Transmitidas por Carrapatos/veterinária , Anaplasma phagocytophilum/isolamento & purificação , Animais , Animais Domésticos , Animais Selvagens , Técnicas de Tipagem Bacteriana , Sequência de Bases , Análise por Conglomerados , Reservatórios de Doenças/microbiologia , Ehrlichiose/microbiologia , França , Loci Gênicos/genética , Variação Genética , Geografia , Humanos , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Doenças Transmitidas por Carrapatos/microbiologia
14.
J Virol ; 88(10): 5870-2, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24574409

RESUMO

Mice overexpressing the prion protein (PrP) sequence from various host species are widely used for measuring infectious titers in prion disease. However, the impact that the transgene expression level might have on the susceptibility to infection raises some concerns about the final biological relevance of these models. Here we report that endpoint titration of a sheep scrapie isolate in sheep and in mice overexpressing the ovine PrP results in similar estimates of the infectious titer.


Assuntos
Suscetibilidade a Doenças , Expressão Gênica , Proteínas PrPSc/biossíntese , Doenças Priônicas/genética , Animais , Camundongos , Proteínas Recombinantes/biossíntese , Ovinos
15.
Emerg Infect Dis ; 20(1): 114-7, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24377668

RESUMO

We report the presence of infectivity in erythrocytes, leukocytes, and plasma of 1 person with variant Creutzfeldt-Jakob disease and in the plasma of 2 in 4 persons whose tests were positive for sporadic Creutzfeldt-Jakob disease. The measured infectivity levels were comparable to those reported in various animals with transmissible spongiform encephalopathies.


Assuntos
Síndrome de Creutzfeldt-Jakob/metabolismo , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Bovinos , Síndrome de Creutzfeldt-Jakob/sangue , Síndrome de Creutzfeldt-Jakob/patologia , Modelos Animais de Doenças , Eritrócitos/metabolismo , Humanos , Leucócitos/metabolismo , Camundongos , Camundongos Transgênicos , Príons/genética , Príons/metabolismo , Príons/patogenicidade
16.
Vet Rec Open ; 1(1): e000036, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-26392876

RESUMO

OBJECTIVE: The present study aimed to estimate and compare the economic impact of Schmallenberg virus (SBV) in different sheep production holdings using partial budget and gross margin analyses in combination with production models. PARTICIPANTS: The sheep production types considered were lowland spring lambing, upland spring lambing and early lambing flocks in the UK, and grass lamb flocks of the Centre and West of France, extensive lambing flocks and dairy sheep flocks in France. METHODOLOGY: Two disease scenarios with distinct input parameters associated with reproductive problems were considered: low and high impact. Sensitivity analyses were performed for the most uncertain input parameters, and the models were run with all of the lowest and highest values to estimate the range of disease impact. RESULTS: The estimated net SBV disease cost per year and ewe for the UK was £19.65-£20.85 for the high impact scenario and £6.40-£6.58 for the low impact scenario. No major differences were observed between the different production types. For France, the net SBV disease cost per year and ewe for the meat sheep holdings was £15.59-£17.20 for the high impact scenario and £4.75-£5.26 for the low impact scenario. For the dairy sheep, the costs per year and ewe were £29.81 for the high impact scenario and £10.34 for the low impact scenario. CONCLUSIONS: The models represent a useful decision support tool for farmers and veterinarians who are facing decisions regarding disease control measures. They allow estimating disease impact on a farm accounting for differing production practices, which creates the necessary basis for cost effectiveness analysis of intervention strategies, such as vaccination.

17.
J Virol ; 88(5): 2406-13, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24284317

RESUMO

In goats, several field studies have identified coding mutations of the gene encoding the prion protein (I/M142, N/D146, S/D146, R/Q211, and Q/K222) that are associated with a lower risk of developing classical scrapie. However, the data related to the levels of resistance to transmissible spongiform encephalopathies (TSEs) of these different PRNP gene mutations are still considered insufficient for developing large-scale genetic selection against scrapie in this species. In this study, we inoculated wild-type (WT) PRNP (I142R154R211Q222) goats and homozygous and/or heterozygous I/M142, R/H154, R/Q211, and Q/K222 goats with a goat natural scrapie isolate by either the oral or the intracerebral (i.c.) route. Our results indicate that the I/M142 PRNP polymorphism does not provide substantial resistance to scrapie infection following intracerebral or oral inoculation. They also demonstrate that H154, Q211, and K222 PRNP allele carriers are all resistant to scrapie infection following oral exposure. However, in comparison to WT animals, the H154 and Q211 allele carriers displayed only moderate increases in the incubation period following i.c. challenge. After i.c. challenge, heterozygous K222 and a small proportion of homozygous K222 goats also developed the disease, but with incubation periods that were 4 to 5 times longer than those in WT animals. These results support the contention that the K222 goat prion protein variant provides a strong but not absolutely protective effect against classical scrapie.


Assuntos
Predisposição Genética para Doença , Doenças das Cabras/genética , Scrapie/genética , Alelos , Animais , Códon , Feminino , Genótipo , Doenças das Cabras/metabolismo , Doenças das Cabras/patologia , Cabras/genética , Proteínas PrPSc/genética , Proteínas PrPSc/metabolismo , Distribuição Tecidual
18.
PLoS One ; 8(1): e54911, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23372789

RESUMO

Small ruminant post-mortem testing programs were initially designed for monitoring the prevalence of prion disease. They are now considered as a potential alternative to genetic selection for eradicating/controlling classical scrapie at population level. If such policy should be implemented, its success would be crucially dependent on the efficiency of the surveillance system used to identify infected flocks. In this study, we first determined the performance of post-mortem classical scrapie detection in eight naturally affected goat herds (total n = 1961 animals) according to the age at culling. These results provided us with necessary parameters to estimate, through a Monte Carlo simulation model, the performance of scrapie detection in a commercial population. According to this model, whatever the number of tests performed, post mortem surveillance will have limited success in identifying infected herds. These data support the contention that scrapie eradication programs relying solely on post mortem testing in goats will probably fail. Considering the epidemiological and pathological similarities of scrapie in sheep and goats, the efficiency of scrapie surveillance in both species is likely to be similar.


Assuntos
Doenças das Cabras/epidemiologia , Scrapie/epidemiologia , Animais , Ensaio de Imunoadsorção Enzimática , Feminino , Doenças das Cabras/diagnóstico , Cabras , Imuno-Histoquímica , Vigilância da População , Prevalência , Príons , Reprodutibilidade dos Testes , Scrapie/diagnóstico , Sensibilidade e Especificidade
19.
J Gen Virol ; 94(Pt 1): 241-245, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23100359

RESUMO

The PrP gene polymorphisms at codons 142 (I/M), 154 (R/H), 211 (R/Q), 222 (Q/K) and 240 (S/P) and their association with susceptibility to classical scrapie infection were investigated in five French goat herds displaying a high disease prevalence (>10%). On the basis of PrP(Sc) detection in the central nervous system and in various lymphoid tissues, 301 of 1343 goats were found to be scrapie infected. The statistical analyses indicated that while P(240) mutation had no direct impact on scrapie infection risk, the H(154), Q(211) and K(222) mutations were associated with high resistance to scrapie. The M(142) mutated allele was associated with a limited protection level against the disease. These results further reinforce the view that, like in sheep, the control and eradication of classical scrapie through the selection of certain PrP alleles could be envisaged in commercial goat population.


Assuntos
Doenças das Cabras/genética , Príons/genética , Scrapie/genética , Alelos , Animais , Sistema Nervoso Central/imunologia , Sistema Nervoso Central/metabolismo , França/epidemiologia , Predisposição Genética para Doença , Doenças das Cabras/epidemiologia , Doenças das Cabras/imunologia , Cabras , Tecido Linfoide/imunologia , Tecido Linfoide/metabolismo , Mutação , Polimorfismo Genético , Prevalência , Scrapie/epidemiologia , Scrapie/imunologia
20.
Stat Methods Med Res ; 22(3): 243-60, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21632696

RESUMO

Owing to the natural evolution of a disease, several events often arise after a first treatment for the same subject. For example, patients with a primary invasive breast cancer and treated with breast conserving surgery may experience breast cancer recurrences, metastases or death. A certain proportion of subjects in the population who are not expected to experience the events of interest are considered to be 'cured' or non-susceptible. To model correlated failure time data incorporating a surviving fraction, we compare several forms of cure rate frailty models. In the first model already proposed non-susceptible patients are those who are not expected to experience the event of interest over a sufficiently long period of time. The other proposed models account for the possibility of cure after each event. We illustrate the cure frailty models with two data sets. First to analyse time-dependent prognostic factors associated with breast cancer recurrences, metastases, new primary malignancy and death. Second to analyse successive rehospitalizations of patients diagnosed with colorectal cancer. Estimates were obtained by maximization of likelihood using SAS proc NLMIXED for a piecewise constant hazards model. As opposed to the simple frailty model, the proposed methods demonstrate great potential in modelling multivariate survival data with long-term survivors ('cured' individuals).


Assuntos
Neoplasias da Mama/patologia , Neoplasias Colorretais/patologia , Modelos Teóricos , Metástase Neoplásica , Recidiva Local de Neoplasia , Readmissão do Paciente , Taxa de Sobrevida , Feminino , Humanos , Funções Verossimilhança
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