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1.
Vet Pathol ; : 3009858231209408, 2023 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-37937724

RESUMO

Sarcoids are common mesenchymal neoplasms of horses. Although there are few studies in which sarcoids have been followed over a long period of time, sarcoids are considered locally invasive and have been reported to frequently recur following surgical excision. Currently, no histological features have been identified to predict which sarcoids will recur after excision. The present study comprised 49 sarcoids for which histology sections were available and in which the recurrence status of the case was known. Each sarcoid was excised from a different horse. Overall, 12 of the 49 (24%) sarcoids recurred after surgical excision. Mitotic count (MC), cellularity, necrosis, nuclear pleomorphism, and inflammation of the sarcoids were evaluated histologically. Of these, MC correlated with recurrence. Four of 5 (80%) sarcoids with an MC ≥ 20 in 2.37 mm2 recurred, which was a significantly higher recurrence rate than that of sarcoids with an MC < 20, 8 of 44 cases recurred (18%), P = .0051. Clinical type was also found to correlate with recurrence. Three of 4 (75%) fibroblastic types recurred, which was a significantly higher recurrence rate than that of sarcoids with other clinical types, 9 of 45 cases (18%), P < .001. In addition, univariate Cox regression analysis confirmed fibroblastic type and MC ≥ 20 as significant predictors for recurrence (P = .016 and P = .005, respectively). To the authors' knowledge, this is the first large study examining recurrence rates in sarcoids, and the first time that histological features have been correlated with recurrence.

2.
Front Vet Sci ; 10: 1096554, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37026093

RESUMO

Introduction: Capacity in veterinary epidemiology is critical to detect, respond and control infectious diseases. Laos veterinary service is limited by having only a small number of veterinarians who graduated from overseas. Animal science graduates support the majority of the Laos veterinary services. The veterinary program was established in 2009 at the National University of Laos. We aimed to understand the national veterinary epidemiology capacity and identify gaps and training needs. Method: A cross-sectional online study was conducted in 2021 targeting central (DLF), provincial (PAFO) and district (DAFO) government animal health officers and veterinary/animal science academics (n = 332). The questionnaire addressed skills, experiences and perceived training needs in outbreak investigation, disease surveillance, data management and analysis, epidemiological surveys, One Health, leadership and communication and biosecurity. A descriptive analysis was performed and associations between demographic factors and epidemiological skills were examined. Results and discussion: In total, 205 respondents completed the questionnaire (61.8% response rate). Respondents reported low or no skills and experience in data management and analysis, epidemiological surveys and One Health. In contrast, higher but limited skills and experiences were reported in outbreak investigation, disease surveillance and biosecurity. Previous epidemiology training was primarily associated with stronger experiences in veterinary epidemiology competencies, followed by respondents that had completed a veterinary degree, highlighting the value of the currently available epidemiology training and veterinary-trained personnel in Lao PDR. This study could help inform the Laos government in its policy planning for field veterinary epidemiology capacity and future training.

3.
Prev Vet Med ; 214: 105889, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36906937

RESUMO

Controlling foot-and-mouth disease (FMD) by vaccination requires adequate population coverage and high vaccine efficacy under field conditions. To assure veterinary services that animals have acquired sufficient immunity, strategic post-vaccination surveys can be conducted to monitor the coverage and performance of the vaccine. Correct interpretation of these serological data and an ability to derive exact prevalence estimates of antibody responses requires an awareness of the performance of serological tests. Here, we used Bayesian latent class analysis to evaluate the diagnostic sensitivity and specificity of four tests. A non-structural protein (NSP) ELISA determines vaccine independent antibodies from environmental exposure to FMD virus (FMDV), and three assays measuring total antibodies derived from vaccine antigen or environmental exposure to two serotypes (A, O): the virus neutralisation test (VNT), a solid phase competitive ELISA (SPCE), and a liquid phase blocking ELISA (LPBE). Sera (n = 461) were collected by a strategic post-vaccination monitoring survey in two provinces of Southern Lao People's Democratic Republic (PDR) after a vaccination campaign in early 2017. Not all samples were tested by every assay and each serotype: VNT tested for serotype A and O, whereas SPCE and LPBE tested for serotype O, and only NSP-negative samples were tested by VNT, with 90 of them not tested (missing by study design). These data challenges required informed priors (based on expert opinion) for mitigating possible lack of model identifiability. The vaccination status of each animal, its environmental exposure to FMDV, and the indicator of successful vaccination were treated as latent (unobserved) variables. Posterior median for sensitivity and specificity of all tests were in the range of 92-99 %, except for the sensitivity of NSP (∼66%) and the specificity of LPBE (∼71 %). There was strong evidence that SPCE outperformed LPBE. In addition, the proportion of animals recorded as having been vaccinated that showed a serological immune response was estimated to be in the range of 67-86 %. The Bayesian latent class modelling framework can easily and appropriately impute missing data. It is important to use field study data as diagnostic tests are likely to perform differently on field survey samples compared to samples obtained under controlled conditions.


Assuntos
Doenças dos Bovinos , Vírus da Febre Aftosa , Febre Aftosa , Animais , Bovinos , Febre Aftosa/diagnóstico , Febre Aftosa/epidemiologia , Febre Aftosa/prevenção & controle , Sorogrupo , Teorema de Bayes , Testes Sorológicos/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Vacinação/veterinária , Anticorpos Antivirais , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/prevenção & controle
4.
Transbound Emerg Dis ; 69(5): e1825-e1838, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35294113

RESUMO

Foot-and-mouth disease (FMD) is endemic in Lao People's Democratic Republic (PDR). A risk-based partial vaccination campaign was implemented in Champasak, Savannakhet and Xiengkhouang Provinces in Lao PDR in 2016-2020, which had limited effects on reducing the circulation of FMD virus. The objectives of this study were to measure the socioeconomic effects of (i) a clinical FMD occurrence and (ii) the vaccination campaign on livestock production performance of smallholders in Lao PDR in 2016-2020. Two questionnaire surveys were conducted at the beginning (2016) and the end (2020) of the campaign, involving 1609 smallholders in 160 villages comparing areas with and without FMD control. Mixed-effect multivariable regression models were fitted to measure the association between livestock production parameters at the household level (rates of birth, sale, purchase, and death, sale price, costs, gross gains and net gains) and (i) a clinical FMD occurrence and (ii) the vaccination campaign, while adjusting for village, herd size, province, vaccination status and FMD knowledge. We found no significant difference in annual household-level costs, gains and net gains by due absence of FMD. However, smallholders with FMD had a 52% higher cattle death rate, a 43% lower goat sale rate and a 78% lower pig sale price than those without FMD. Smallholders with FMD also had a lower purchase rate of livestock (cattle -15% and pigs -93%) compared with those without FMD. There was a general improvement in livestock production in target villages over the 5 years of the vaccination campaign, whereas there was an overall decline in non-target villages, resulting in USD 218 higher net gains in target villages in 2020 compared with non-target villages, possibly due to reduced severity of disease and improved awareness on FMD and general livestock health. The findings of this study will help improve strategies for sustainable and efficient FMD control in Lao PDR, while the challenges and importance of gaining participation by smallholders are highlighted.


Assuntos
Doenças dos Bovinos , Vírus da Febre Aftosa , Febre Aftosa , Doenças dos Suínos , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/prevenção & controle , Surtos de Doenças/veterinária , Características da Família , Febre Aftosa/epidemiologia , Febre Aftosa/prevenção & controle , Humanos , Programas de Imunização , Laos/epidemiologia , Gado , Suínos , Doenças dos Suínos/epidemiologia , Vacinação/veterinária
5.
Transbound Emerg Dis ; 69(4): e309-e321, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34412164

RESUMO

Foot and mouth disease (FMD) is endemic in the Lao People's Democratic Republic (PDR) and it causes economic loss to smallholder husbandry systems. An intervention programme based on a risk-based partial vaccination strategy was implemented in three provinces of Lao PDR (Champasak, Savannakhet and Xiangkhouang) to immunise domestic cattle and buffalo during 2016-2020. Two cross-sectional surveys were conducted in 2016/17 and 2020 to evaluate the impact of the vaccination programme on the prevalence of FMD virus exposure and clinical incidence of the disease. A total of 212 villages were visited during the two surveys, collecting 1609 household-level questionnaire results and 5931 blood samples of domestic cattle and buffalo. Blood samples were tested for the presence of antibodies to the non-structural proteins of FMD virus, and seroprevalence of 42.5 and 47.5% in 2016/17 and 2020, respectively were found. Multivariable regression analysis indicated that the efficacy of the FMD vaccination programme for reducing FMD virus circulation varied by province. In general, the incidence of clinical FMD increased toward the end of the 5-year intervention period, coinciding with a reduction of vaccine coverage in the last 2 years of the period. The findings suggest that the risk-based vaccination strategy achieved a marginally protective effect against the circulation of FMD virus with the possible limiting factors being operational constraints of public veterinary services, lack of farmers' compliance and unsustainable funding. We conclude that consistent resource availability and higher vaccination coverage is required to successfully control FMD with a risk-based vaccination strategy in Lao PDR.


Assuntos
Doenças dos Bovinos , Vírus da Febre Aftosa , Febre Aftosa , Animais , Búfalos , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/prevenção & controle , Estudos Transversais , Surtos de Doenças/veterinária , Febre Aftosa/epidemiologia , Febre Aftosa/prevenção & controle , Humanos , Incidência , Laos/epidemiologia , Estudos Soroepidemiológicos , Vacinação/veterinária
6.
Int J Food Microbiol ; 127(1-2): 84-90, 2008 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-18625534

RESUMO

Foot-and-mouth disease virus (FMDV) causes one of the most contagious diseases affecting cloven-hoofed animals (e.g., cattle and swine) and causes severe economic loss for many countries. The resistance to UV irradiation of FMDV strains isolated from outbreaks in Thailand was investigated in phosphate-buffered saline at 25 degrees C. Since the regression coefficients of linear regression were large and root mean square errors were small, UV inactivation could be appropriately summarized and fitted well by a linear equation. The first-order kinetics then could describe UV inactivation, which was experimentally and mathematically shown in this study to be an effective means to inactivate FMDV in suspension. The decimal inactivation dose (DID) was modified from D value in traditional thermal-inactivation kinetics. The DID was the amount of UV irradiation required to reduce the number of microorganisms by a factor of 10, or by 90%. DIDs of FMDV serotypes O189, A132, A-Sakol, and AS1 ranged from 19.66 to 31.31 mWs/cm2. FMDV serotype AS1 was the most UV-resistant, and FMDV serotype A132 was the least UV-resistant. UV resistance of FMDV did not vary significantly among strains and serotypes (P value > 0.05). DID raw data were used to determine the fitted probability distributions by simulation software @Risk. The fitted distributions suggested were Exponential, Logistic, Normal, and LogNormal. Exponential distribution was the best fit by Chi-square test, Kolmogorov-Smirnov test, and Anderson-Darling test (P value >0.10). The parameter beta of the Exponential distribution, equivalent to the mean DID, was 24.173 mWs/cm2 as a first-order model. Poisson probability distribution described the uncertainty of parameter beta for the second-order model. After running simulations of both first-order and second-order models, the curves of both first-order and second-order models were overlaid on the same graph, which indicated that both models were only slightly different. This study concluded that FMDV in suspension was effectively inactivated by UV irradiation, the fitted probability distribution for UV inactivation was Exponential, and source of total uncertainty of this UV-inactivation model was not the uncertainty component.


Assuntos
Vírus da Febre Aftosa/efeitos da radiação , Raios Ultravioleta , Inativação de Vírus/efeitos da radiação , Algoritmos , Relação Dose-Resposta à Radiação , Vírus da Febre Aftosa/crescimento & desenvolvimento , Vírus da Febre Aftosa/fisiologia , Cinética , Modelos Lineares , Sorotipagem , Temperatura , Tailândia
7.
Tuberculosis (Edinb) ; 99: 120-127, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27450013

RESUMO

A study was undertaken to determine whether BCG vaccination of cattle post-challenge could have an effect on a very early Mycobacterium bovis infection. Three groups of calves (n = 12/group) were challenged endobronchially with M. bovis and slaughtered 13 weeks later to examine for tuberculous lesions. One group had been vaccinated prophylactically with BCG Danish vaccine 21 weeks prior to challenge; a second group was vaccinated with a 4-fold higher dose of BCG Danish 3 weeks post-challenge and the third group, remained non-vaccinated. Vaccination prior to challenge induced only minimal protection with just a significant reduction in the lymph node lesion scores. Compared to the non-vaccinated group, BCG vaccination post-challenge produced no reduction in gross pathology and histopathology, but did result in significant increases in mRNA expression of pro-inflammatory mediators (IFN-γ, IL-12p40, IL-17A, IRF-5, CXCL9, CXCL10, iNOs, and TNF-α) in the pulmonary lymph nodes. Although there was no significant differences in the gross pathology and histopathology between the post-challenge BCG and non-vaccinated groups, the enhanced pro-inflammatory immune responses observed in the post-challenge BCG group suggest caution in the use of high doses of BCG where there is a possibility that cattle may be infected with M. bovis prior to vaccination.


Assuntos
Vacina BCG/administração & dosagem , Citocinas/imunologia , Mediadores da Inflamação/imunologia , Pulmão/imunologia , Linfonodos/imunologia , Mycobacterium bovis/imunologia , Tuberculose Bovina/tratamento farmacológico , Tuberculose Pulmonar/tratamento farmacológico , Animais , Vacina BCG/toxicidade , Bovinos , Citocinas/genética , Citocinas/metabolismo , Interações Hospedeiro-Patógeno , Esquemas de Imunização , Mediadores da Inflamação/metabolismo , Testes de Liberação de Interferon-gama , Pulmão/metabolismo , Pulmão/microbiologia , Pulmão/patologia , Linfonodos/metabolismo , Linfonodos/microbiologia , Linfonodos/patologia , Masculino , Mycobacterium bovis/patogenicidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Tempo , Teste Tuberculínico , Tuberculose Bovina/imunologia , Tuberculose Bovina/metabolismo , Tuberculose Bovina/microbiologia , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/metabolismo , Tuberculose Pulmonar/microbiologia , Regulação para Cima
8.
PLoS One ; 11(7): e0159861, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27472482

RESUMO

Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2) formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation). A control group of sheep (n = 6) was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1-2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible.


Assuntos
Anticorpos/análise , Proteínas Arqueais/administração & dosagem , Rúmen/microbiologia , Saliva/imunologia , Carneiro Doméstico/imunologia , Vacinação/métodos , Animais , Anticorpos/imunologia , Antígenos/imunologia , Archaea/isolamento & purificação , Proteínas Arqueais/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Imunoglobulina A/análise , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Imunoglobulina G/análise , Imunoglobulina G/sangue , Imunoglobulina G/imunologia
9.
Vet Immunol Immunopathol ; 164(3-4): 201-7, 2015 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-25782351

RESUMO

Methane is produced in the rumen of cattle by a group of archaea (single-celled organisms forming a domain distinct from bacteria and eucarya) called methanogens. Vaccination against methanogens has the potential to reduce methane emissions by inducing antibodies in saliva which are transferred to the rumen and diminish the ability of methanogens to produce methane. Since it is likely that an effective vaccination strategy will need to produce high levels of methanogen-specific antibody in the saliva; the choice of adjuvant, route of vaccination and stability of saliva-derived antibody in the rumen all need to be considered. In this study, stability of IgA and IgG in rumen fluid was determined using an in vitro assay. IgA levels in cattle saliva were reduced by only 40% after 8h exposure to rumen contents while IgG levels were reduced by 80%. These results indicated that antibody is relatively stable in the bovine rumen. A trial was conducted in cattle to investigate induction of immune responses to a methanogen protein, recombinant glycosyl transferase protein (rGT2) from Methanobrevibacter ruminantium M1. Groups of cattle (n=6) were vaccinated subcutaneously with rGT2, formulated with Montanide ISA61 with or without the TLR4 agonist, monophosphoryl lipid A (MPL). A control group (n=6) was not vaccinated. Strong antigen-specific IgG and moderate IgA responses were measured in the serum and saliva of the vaccinated animals and antibody was also detected in the rumen.


Assuntos
Anticorpos Antiarchaea/biossíntese , Proteínas Arqueais/imunologia , Glicosiltransferases/imunologia , Methanobrevibacter/imunologia , Rúmen/imunologia , Saliva/imunologia , Vacinação/veterinária , Animais , Bovinos , Imunoglobulina A/análise , Imunoglobulina G/análise , Masculino
10.
Vet Immunol Immunopathol ; 160(1-2): 51-60, 2014 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-24768453

RESUMO

Streptococcus uberis is a major cause of mastitis in dairy cows worldwide and currently, there is no vaccine commercially available against this form of mastitis. In the current study, cell-free extracts (CFE) were prepared from each of three different S. uberis strains, designated as #3, #24 and #363 representative of the three main sequence types of S. uberis that cause mastitis in New Zealand. These proteins were formulated into vaccines with Emulsigen-D and the immunogenicity of the vaccines was determined in both calves and dairy cows. Two groups of calves (n=5/group) were vaccinated subcutaneously with CFE from strain #24 or strains #3, #24 and #363 formulated with Emulsigen-D, respectively. A third group (n=5) was vaccinated with CFE from the three strains formulated with Emulsigen-D and also containing recombinant bovine granulocyte macrophage colony-stimulating factor while, a control group (n=5) was not vaccinated. Vaccinated animals produced strong antibody responses to the S. uberis antigens and an antigen-specific cytotoxic effect against blood monocytes/macrophages that had phagocytosed S. uberis, with no significant differences in responses observed between the three vaccinated groups. In a second trial, the safety and immunogenicity of the vaccine containing CFE from all three strains of S. uberis and Emulsigen-D was determined in dairy cows. A group of six cows were vaccinated subcutaneously at 3 and 1 week prior to dry off and revaccinated 2-3 weeks before calving. Immune responses in blood and mammary gland secretions (MGS) were monitored during the dry period and in the subsequent lactation. The vaccine was well tolerated with no adverse effect from vaccination observed in any of the cows. Vaccination induced an antigen-specific cytotoxic effect against blood monocytes/macrophages that had phagocytosed S. uberis, moderate antigen-specific IFN-γ responses in blood and strong antibody responses in both blood and MGS. In conclusion, the results suggest vaccination of cattle with S. uberis CFE by the subcutaneous route can induce both cellular and humoral responses.


Assuntos
Antígenos de Bactérias/imunologia , Citocinas/metabolismo , Mastite Bovina/prevenção & controle , Vacinas Estreptocócicas/imunologia , Streptococcus/imunologia , Linfócitos T Citotóxicos/fisiologia , Animais , Bovinos , Citocinas/genética , Indústria de Laticínios , Feminino , Infecções Estreptocócicas/prevenção & controle , Infecções Estreptocócicas/veterinária
11.
PLoS One ; 9(9): e106519, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25180583

RESUMO

In both humans and animals, controversy exists concerning the duration of protection induced by BCG vaccine against tuberculosis (TB) and whether revaccination enhances protection. A long-term study was undertaken to determine whether BCG-vaccinated calves would be protected against challenge with Mycobacterium bovis 2½ years after vaccination and to determine the effect of revaccination after 2 years. Seventy-nine calves were divided into five groups (n = 15-17 calves/group) with four of the groups vaccinated subcutaneously with 105 CFU of BCG Danish at 2-4 weeks of age and the fifth group serving as non-vaccinated controls. Three of the four BCG-vaccinated groups were revaccinated 2 years after the initial vaccination. One BCG-vaccinated group was revaccinated with BCG. A second group was vaccinated subcutaneously with a TB protein vaccine consisting of biopolyester particles (Biobeads) displaying two mycobacterial proteins, ESAT-6 and Antigen 85A, mixed with an adjuvant. A third group was vaccinated with TB proteins from M. bovis culture filtrate, mixed with an adjuvant. Twenty-three weeks after the BCG revaccination, all animals were challenged endotracheally with virulent M. bovis and a further 13 weeks later, animals were killed and necropsied to determine protection against TB. The BCG-vaccinated animals produced positive tuberculin caudal fold intradermal (15 of 62 animals) and IFN-γ TB test responses (six of 62 animals) at 6 months after vaccination, but not at subsequent time-points compared to the non-vaccinated animals. Calves receiving a single vaccination with BCG vaccine 2½ years prior to challenge were not protected against TB, while those revaccinated with BCG 2 years after the initial vaccination displayed significant reductions in lung and pulmonary lymph node lesion scores compared to the non-vaccinated animals. In contrast, no reduction in lesion scores was observed in the animals revaccinated with the TB protein vaccines with their immune responses biased towards induction of antibody.


Assuntos
Vacina BCG/administração & dosagem , Imunização Secundária/veterinária , Mycobacterium bovis/imunologia , Tuberculose Bovina/imunologia , Tuberculose Bovina/prevenção & controle , Animais , Anticorpos Antibacterianos/sangue , Bovinos , Feminino , Interferon gama/sangue , Pulmão/patologia , Linfonodos/patologia , Mycobacterium bovis/patogenicidade , Fatores de Tempo , Teste Tuberculínico , Vacinas contra a Tuberculose/administração & dosagem , Tuberculose Bovina/diagnóstico
12.
Vet Immunol Immunopathol ; 145(1-2): 471-8, 2012 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-22078656

RESUMO

Johne's disease caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic enteric disease of cattle. The mechanism how MAP can co-exist in the gastro-intestinal tract despite a massive infiltration of immune cells is not known. Toll-like receptors (TLRs) are known to play an important role in both innate and acquired immune responses but it is unclear what role different TLRs play in response to MAP. In this study, 38 cull cows from herds infected with MAP were classified into four groups, based on MAP culture from gut tissues and histopathological lesion scores. The expression of TLR1, 2 and 4 mRNA from MAP antigen-stimulated mesenteric lymph node (MLN) cultures and peripheral blood mononuclear cells (PBMCs) and in the MLN and ileum tissues of these animals was determined. MAP antigen-specific expression of TLR1 in MLN and PBMC was significantly lower in the MAP-infected groups than the non-infected control group, suggesting that in MAP-infected animals there is impairment in the up-regulation of TLR1 in response to MAP antigen. TLR4 expression in MLN tissues was significantly higher in the severely infected group than the control group suggesting up-regulation of endogenous TLR4 expression at a site of MAP infection in animals severely affected with Johne's disease. A preliminary screening of TLR1, 2 and 4 in the cull cows revealed the presence of polymorphisms in TLR1 and TLR2. In summary, one mechanism how MAP may subvert the immune system is that there is an apparent lack of recognition of MAP antigens as foreign by TLR1 in MAP-infected cows.


Assuntos
Doenças dos Bovinos/imunologia , Mycobacterium avium subsp. paratuberculosis , Paratuberculose/imunologia , Receptores Toll-Like/biossíntese , Animais , Bovinos , Doenças dos Bovinos/genética , Doenças dos Bovinos/microbiologia , Feminino , Regulação da Expressão Gênica/imunologia , Mycobacterium avium subsp. paratuberculosis/imunologia , Paratuberculose/genética , Filogenia , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de DNA/veterinária , Receptores Toll-Like/genética
13.
Vet Immunol Immunopathol ; 149(3-4): 225-36, 2012 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-22871577

RESUMO

This study examined the immune responses related to the infection, progression and control of Mycobacterium avium subsp. paratuberculosis (MAP) infection in calves. Twenty calves were challenged orally with MAP and 11 non-challenged calves served as controls. Approximately half the calves from each group were sacrificed at either 7 or 15 months post-challenge (PC). The majority of the challenged calves (19/20) shed MAP in feces 2-4 months PC, but thereafter fecal shedding reduced markedly. The severity of infection was reduced at 15 months PC compared to that at 7 months PC as seen from a significantly lower isolation of MAP from tissues and lower lesion scores (P<0.05). In addition, there was a reduction in the upregulation of gene expression of gamma interferon, interleukin-10 (IL-10) and inducible nitric oxide synthase from the antigen-stimulated mesenteric lymph node (MLN) cultures of the challenged calves. No evidence of infection was detected in the control calves. The severity of the infection in individual calves at 15 months PC as indicated from the number of tissue culture positive sites, was negatively related to IL-10 released from antigen-stimulated peripheral blood mononuclear cells (P<0.05). Collectively the data indicated that the severity of the MAP infection was reduced in the calves at 15 months PC and in a specific time period during infection, IL-10 may play a role in reducing the severity of this disease.


Assuntos
Doenças dos Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Mycobacterium avium subsp. paratuberculosis/imunologia , Paratuberculose/imunologia , Paratuberculose/microbiologia , Animais , Bovinos , Fezes/microbiologia , Feminino , Interferon gama/imunologia , Interleucina-10/imunologia , Linfonodos/imunologia , Linfonodos/microbiologia , Mycobacterium avium subsp. paratuberculosis/genética , Óxido Nítrico Sintase Tipo II/imunologia , Reação em Cadeia da Polimerase/veterinária , RNA Mensageiro/química , RNA Mensageiro/genética , Distribuição Aleatória , Estatísticas não Paramétricas
14.
J Vet Diagn Invest ; 23(4): 743-52, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21908317

RESUMO

A SYTO9 real-time polymerase chain reaction assay for detection of pathogenic Leptospira spp. based on amplification of DNA gyrase subunit B (gyrB) gene has been optimized and evaluated for sensitivity and specificity on kidney and urine samples of New Zealand farmed deer. The detection limit was 10(3) cells/ml (2-10 copies/reaction). Comparison of the assay on deer kidneys (n = 268) with culture as the gold standard revealed a sensitivity and specificity of 85% and 99.2%, respectively. For deer urine (n = 113), the assay was compared with known inoculated samples and revealed a sensitivity and specificity of 96.7% and 100%, respectively. The assay was applied for quantifying pathogenic leptospires shed naturally in deer urine and revealed a detectable concentration of 3.7 × 10(3) to 1.7 × 10(6) cells/ml. To assess the assay's capability for identifying pathogenic Leptospira spp., 14 field isolates of L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona were amplified for polymerase chain reaction (PCR) product, purified, and sequenced. When compared with the National Center for Biotechnology Information database, sequence data matched with L. borgpetersenii serovar Hardjo-bovis in 13 samples and L. interrogans serovar Pomona in 1 sample, which was consistent with the microscopic agglutination test (MAT). Sequence analysis of purified PCR product amplified directly from kidney and urine samples also yielded serovar-comparable MAT results. Results suggest that the assay is rapid, sensitive, and specific for detection of pathogenic leptospires in deer clinical samples. The developed assay can also be used for estimating the concentration of leptospires and identifying Leptospira spp. in combination with DNA sequencing.


Assuntos
Cervos , Rim/microbiologia , Leptospira/isolamento & purificação , Leptospirose/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Cervos/urina , Leptospira/classificação , Leptospirose/epidemiologia , Leptospirose/microbiologia , Leptospirose/urina , Nova Zelândia/epidemiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade
15.
Clin Vaccine Immunol ; 18(9): 1467-76, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21795461

RESUMO

Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease, is able to dampen or distort immune responses at the mucosal sites and coexist with a massive infiltration of immune cells in the gastrointestinal tract. Knowledge of the mechanism by which M. avium subsp. paratuberculosis subverts the immune response at the mucosal level in cattle is important for the development of improved disease control strategies, including new vaccines and diagnostic tests. In this study, 38 cull cows from herds infected with M. avium subsp. paratuberculosis were divided into four groups, based on M. avium subsp. paratuberculosis culture from gut tissues and histopathological lesion scores. Cytokine gene expression and secretion from M. avium subsp. paratuberculosis sonicate-stimulated peripheral blood mononuclear cell (PBMC) and mesenteric lymph node (MLN) cultures of the animals were compared. Antigen stimulation of MLN cells from the severely lesioned group resulted in significant upregulation of the mRNA expression of five cytokines, gamma interferon (IFN-γ), interleukin-10 (IL-10), IL-13, IL-17A, and tumor necrosis factor alpha (TNF-α), which have a diverse range of functions, while there was no significant upregulation of these cytokines by the other groups. There were major differences between the responses of the PBMC and MLN cultures, with higher levels of secreted IFN-γ released from the MLN cultures and, conversely, higher levels of IL-10 released from the PBMC cultures. The upregulation of all five cytokines from cells at the site of infection in the severely lesioned animals suggested a dysregulated immune response, contributing to a failure to clear infection in this group of animals.


Assuntos
Citocinas/metabolismo , Linfonodos/citologia , Linfonodos/imunologia , Mesentério/imunologia , Mycobacterium avium subsp. paratuberculosis/imunologia , Paratuberculose/imunologia , Paratuberculose/fisiopatologia , Animais , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/patologia , Doenças dos Bovinos/fisiopatologia , Células Cultivadas , Feminino , Valva Ileocecal/patologia , Íleo/patologia , Leucócitos Mononucleares/imunologia , Linfonodos/patologia , Mesentério/patologia , Mycobacterium avium subsp. paratuberculosis/patogenicidade , Paratuberculose/microbiologia , Paratuberculose/patologia , Índice de Gravidade de Doença , Regulação para Cima
16.
Appl Environ Microbiol ; 73(22): 7177-84, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17660312

RESUMO

The heat resistance of foot-and-mouth disease virus (FMDV) strains isolated from outbreaks in Thailand was investigated in phosphate-buffered saline (PBS) at 50, 60, 70, 80, 90, and 100 degrees C. The first-order kinetic model fitted most of the observed linear inactivation curves. The ranges of decimal-reduction time (D value) of FMDV strains at 50, 60, 70, 80, 90, and 100 degrees C were 732 to 1,275 s, 16.37 to 42.00 s, 6.06 to 10.87 s, 2.84 to 5.99 s, 1.65 to 3.18 s, and 1.90 to 2.94 s, respectively. The heat resistances of FMDV strains at lower temperature (50 degrees C) were not serotype specific. The effective inactivating temperature is approximately 60 degrees C. Heat resistances of FMDV strains at 90 and 100 degrees C were not statistically different (P > 0.05), while the FMDV serotype O (OPN) appeared to be the most heat resistant at 60 to 80 degrees C. The other observed inactivation curves were linear with shoulder or tailing (biphasic curves). The shoulder effect was mostly observed at 90 and 100 degrees C, while the tailing effect was mostly observed at 50 to 80 degrees C. The adjusted D values in the case of shoulder and tailing effects did not affect the overall estimated heat resistance of these FMDV strains, so even unadjusted D values of deviant inactivation curves were legitimate. The z values of FMDV serotypes O, A, and Asia 1 were 21.78 to 23.26, 20.75 to 22.79, and 19.87 degrees C, respectively. The z values of FMDV strains studied were not statistically significantly different (P > 0.05). The results of this study indicated that the heat resistance in PBS of FMDV strains from Thailand was much less than had been reported for foreign epidemic FMDV strains.


Assuntos
Vírus da Febre Aftosa/crescimento & desenvolvimento , Temperatura Alta , Adaptação Fisiológica , Algoritmos , Vírus da Febre Aftosa/classificação , Temperatura , Tailândia
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