Infectious Disease Of Ruminants Research Articles

Epithelial processed Mycobacterium avium subsp. paratuberculosis induced prolonged Th17 response and suppression of phagocytic maturation in bovine peripheral blood mononuclear cells

Johne’s disease (JD) caused by Mycobacterium avium subsp. paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. Understanding the mechanism of persistency of MAP is key to produce novel ideas for the development of new diagnostic methods or prevention techniques. We sought interactions between the host and MAP using epithelial passage model, which mimic initial stage of infection. From the transcriptomic analysis of bovine immune cells (PBMCs), it was suggested that infection through the epithelial cells elicited prolonged Th17-derived immune response, as indicated by upregulation of IL-17A, IL-17F and RORC until 120 h p.i., compared to directly infected PBMCs. Global downregulation of gene expression was observed after 72 h p.i., especially for genes encoding cell surface receptors of phagocytic cells, such as Toll-like receptors and MHC class II molecules. In addition, the cholesterol efflux transporters ABCA1, ABCG1, and APOE, which are regulated by the LXR/RXR pathway, were downregulated. In summary, it would be suggested that the host initiate immune response to activate Th17-derived cytokines, and MAP survives persistently by altering the host adaptive immune response by suppressing surface receptors and manipulating lipid metabolism in phagocytic cells.

Comparative analysis of serological tests and fecal detection in the diagnosis of Mycobacterium avium subspecies paratuberculosis infection

Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. The most effective way to eradicate JD is to detect infected individuals as early as possible and remove them from the herd. However, it is difficult to detect infected individuals early with the currently using diagnostic methods. Two serological diagnostic kits commercially used worldwide and a fecal detection test were compared using 298 serum samples and feces of cattle in this study to present an efficient diagnostic method. Although there was a high correlation between the 2 serological diagnostic kits (R2 = 0.7473), kit A showed a higher serological positive rate. However, the correlation between fecal tests and serological diagnosis was very low. MAP was also detected in fecal tests in many serologically negative individuals. In the periodical diagnosis of JD, MAP was detected in the feces of only cows with the higher antibody titer to MAP. These results suggest that for effective eradication of JD, early detection of infected individuals by fecal tests together with the serological tests currently in use and by removal of infected individuals are needed. Keywords: paratuberculosis, diagnosis, enzyme-linked immunosorbent assay, polymerase chain reaction

Immunoinformatic-Based Prediction of Candidate Epitopes for the Diagnosis and Control of Paratuberculosis (Johne's Disease).

Paratuberculosis is an infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). MAP is an intracellular pathogen with a possible zoonotic potential since it has been successfully isolated from the intestine and blood of Crohn’s disease patients.Since no cure is available, after the detection of the disease, animal culling is the sole applicable containment strategy. However, the difficult detection of the disease in its subclinical form, facilitates its spread raising the need for the development of effective diagnosis and vaccination strategies. The prompt identification and isolation of the infected animals in the subclinical stage would prevent the spread of the infection.In the present study, an immunoinformatic approach has been used to investigate the immunogenic properties of 10 MAP proteins. These proteins were chosen according to a previously published immunoproteomics approach. For each previously-described immunoreactive protein, we predicted the epitopes capable of eliciting an immune response by binding both B-cells and/or class I MHC antigens. The retrieved peptide sequences were analyzed for their specificity and cross-reactivity. The final aim is to employ the discovered peptides sequences as a filtered library useful for early-stage diagnosis and/or to be used in novel multi-subunit or recombinant vaccine formulations.

Genetic Resistance to Johne's Disease in Cattle

Paratuberculosis (Johne's disease) is an economically significant, infectious disease of ruminants caused by Mycobacterium avium subsp paratuberculosis, and characterized by progressive weight loss and nonresponsive diarrhea. Genetic factors have been associated with differences in host susceptibility to bovine paratuberculosis. Research has been aimed at detecting associations between susceptibility and polymorphisms at candidate genes with no definitive results. Several gene products are candidates for this association. Interferon gamma is an inducible cytokine with a crucial role in the innate host response to intracellular bacteria. Toll-like receptors are trans-membrane structures responsible for coordination of innate and adaptive immune responses. The SLC11A1 gene plays an important role in innate immunity, preventing bacterial growth in macrophages during the initial stages of infection. The objective of this candidate gene case-control study was to characterize the distribution of polymorphisms in three candidate genes related to the immune function; interferon gamma (BoIFNG), toll-like receptor 4 (TLR4), and solute carrier family 11 member1 (SLC11A1) genes, and test their association with paratuberculosis infection in cattle.

An efficient production of hybrid recombinant protein comprising non-structural proteins (NS 1 & NS 3) of bluetongue virus in prokaryotic expression system

Strategic design and suitable purification techniques are of paramount importance in the production of recombinant proteins, if intended for use in a diagnostic assay. However, there is no single protocol that can be universally adopted for obtaining proteins in requisite quality and quantity across various platforms. In this study, we have targeted proteins of bluetongue virus (BTV), which is the causative agent of an arthropod-borne infectious disease in ruminants. Traditionally, serological diagnosis of the disease has rested upon either virus neutralization test or on an ELISA test that employed a recombinant structural (VP1, VP7) protein. Among the non-structural (NS) proteins of BTV, NS1 and NS3, are preferred candidate antigens in development of immuno-diagnostics as these provide the option for identifying recent/ongoing infection. However, the difficulty in production/purification of recombinant full length NS proteins of BTV in sufficient quantity and quality in various expression systems, due to inherent structural complexities, have restricted their wider applicability as immunodiagnostic reagents. To circumvent the difficulties associated with production/purification, we developed a novel NS1 and NS3 fusion gene (∼1302 bp) encoding for NS1 N-terminus (1M to G252 aa) and NS3 protein containing the N- and C-termini with a deletion of two hydrophobic domains along with intervening variable central domain (118A to A182 aa) of bluetongue virus 23. This construct was cloned, over-expressed and efficiently purified by single step affinity chromatography under unique denaturing/renaturing condition. The purified fusion protein was found suitable for detection of antibodies against BTV in an indirect ELISA (iELISA).

Johne’s disease control: a challenging yet achievable goal

Johne’s disease is an infectious disease of ruminants caused by the bacterium Mycobacterium avium subspecies paratuberculosis (Map). It is found globally and is undoubtedly one of the most challenging infectious...

Multiple genome segments determine virulence of bluetongue virus serotype 8.

ABSTRACTBluetongue virus (BTV) causes bluetongue, a major hemorrhagic disease of ruminants. In order to investigate the molecular determinants of BTV virulence, we used a BTV8 strain minimally passaged in tissue culture (termed BTV8L in this study) and a derivative strain passaged extensively in tissue culture (BTV8H) in in vitro and in vivo studies. BTV8L was pathogenic in both IFNAR−/− mice and in sheep, while BTV8H was attenuated in both species. To identify genetic changes which led to BTV8H attenuation, we generated 34 reassortants between BTV8L and BTV8H. We found that partial attenuation of BTV8L in IFNAR−/− mice was achieved by simply replacing genomic segment 2 (Seg2, encoding VP2) or Seg10 (encoding NS3) with the BTV8H homologous segments. Fully attenuated viruses required at least two genome segments from BTV8H, including Seg2 with either Seg1 (encoding VP1), Seg6 (encoding VP6 and NS4), or Seg10 (encoding NS3). Conversely, full reversion of virulence of BTV8H required at least five genomic segments of BTV8L. We also demonstrated that BTV8H acquired an increased affinity for glycosaminoglycan receptors during passaging in cell culture due to mutations in its VP2 protein. Replication of BTV8H was relatively poor in interferon (IFN)-competent primary ovine endothelial cells compared to replication of BTV8L, and this phenotype was determined by several viral genomic segments, including Seg4 and Seg9. This study demonstrated that multiple viral proteins contribute to BTV8 virulence. VP2 and NS3 are primary determinants of BTV pathogenesis, but VP1, VP5, VP4, VP6, and VP7 also contribute to virulence.IMPORTANCE Bluetongue is one of the major infectious diseases of ruminants, and it is listed as a notifiable disease by the World Organization for Animal Health (OIE). The clinical outcome of BTV infection varies considerably and depends on environmental and host- and virus-specific factors. Over the years, BTV serotypes/strains with various degrees of virulence (including nonpathogenic strains) have been described in different geographical locations. However, no data are available to correlate the BTV genotype to virulence. This study shows that BTV virulence is determined by different viral genomic segments. The data obtained will help to characterize thoroughly the pathogenesis of bluetongue. The possibility to determine the pathogenicity of virus isolates on the basis of their genome sequences will help in the design of control strategies that fit the risk posed by new emerging BTV strains.

Prevalence and pathological study of Paramphistomum infection in the small intestine of slaughtered ovine.

Paramphistomiasis, a trematode infectious disease in ruminants, has been neglected but has recently emerged as an important cause of productivity loss. The small intestine of slaughtered sheep was collected weekly from abattoirs (Kermanshah, Sanandaj, Tabriz and Urmia Slaughterhouses) to monitoring the seasonal occurrence of Paramphistomosis, 2,421 sheep carcasses (743 male (30.69%) and 1,678 female (69.31%)) were examined, out of which 0.041% were positive for Paramphistomum infestation. Furthermore, upon evaluation Paramphistomum termatodes, Gastrothylax crumenifer and Cotylophoron detected as well. Overall, the small intestinal infestation by such parasite was 0.041% which contained hyperemia, severe congestion and haemorrhage. The highest infection in the sheep infected with Paramphistomum spp. was found during the summer (July to August) (6.7, 2%) and followed by the autumn seasons (November to October) (3.8, 2.3%). Microscopic study of the small intestine revealed dilatation of intestinal glands, destruction of superficial glands, replacement of fibrin, diffuse infiltration of inflammatory cells and fibrinonecrotic enteritis. Other changes as congestion hemorrhage and nodules of Ostertagia were observed in total examination of small intestines. According to statistical analysis by SPSS software and Chi square test revealed that there is significant difference between pathologic changes, seasons and ecological situations of the region (p<0.05). There was no significant difference between age, gender and sample pH of examined sheep (p>0.05).According to the results of pathologic changes of sheep small intestines, preventive measurements in the area should be taken to decrease the damages, so applying a parasitic control program is recommended.

Recovery rates of bluetongue virus serotypes 1, 2, 4 and 8 Spanish strains from orally infected Culicoides imicola in South Africa

Bluetongue (BT) is an infectious disease of ruminants that has spread northwards in Europe during the last decade. The aetiological agent of the disease is an arbovirus [bluetongue virus (BTV)] that belongs to the genus Orbivirus (family Reoviridae). The virus is transmitted by certain species of biting midge within the genus Culicoides (Diptera: Ceratopogonidae). Information on the vector status of the Culicoides species in a specific area will be essential to predict the risk for BTV incursion. Field-collected Culicoides (Avaritia) imicola Kieffer from South Africa were fed on blood containing several Spanish isolates of BTV. Despite the high virus concentrations in the bloodmeal (5.1-6.4 log(10) TCID(50) /mL of blood), virus was recovered from <1% of midges assayed after incubation. Virus concentrations >2.5 log(10) TCID(50) /midge in individual infected C. imicola suggest virus replication with possible risk for transmission to susceptible vertebrate hosts in the field for at least two of the serotypes assayed (BTV-1 and BTV-2). A third serotype (BTV-4) was very close to the estimated threshold for transmission. The relatively low to near refractory status of C. imicola compared with other vector species such as Culicoides bolitinos supports previous results, indicating that Culicoides species other than C. imicola may play a more important role in the epidemiology of BTV.

Estimating front-wave velocity of infectious diseases: a simple, efficient method applied to bluetongue

Understanding the spatial dynamics of an infectious disease is critical when attempting to predict where and how fast the disease will spread. We illustrate an approach using a trend-surface analysis (TSA) model combined with a spatial error simultaneous autoregressive model (SARerr model) to estimate the speed of diffusion of bluetongue (BT), an infectious disease of ruminants caused by bluetongue virus (BTV) and transmitted by Culicoides. In a first step to gain further insight into the spatial transmission characteristics of BTV serotype 8, we used 2007-2008 clinical case reports in France and TSA modelling to identify the major directions and speed of disease diffusion. We accounted for spatial autocorrelation by combining TSA with a SARerr model, which led to a trend SARerr model. Overall, BT spread from north-eastern to south-western France. The average trend SARerr-estimated velocity across the country was 5.6 km/day. However, velocities differed between areas and time periods, varying between 2.1 and 9.3 km/day. For more than 83% of the contaminated municipalities, the trend SARerr-estimated velocity was less than 7 km/day. Our study was a first step in describing the diffusion process for BT in France. To our knowledge, it is the first to show that BT spread in France was primarily local and consistent with the active flight of Culicoides and local movements of farm animals. Models such as the trend SARerr models are powerful tools to provide information on direction and speed of disease diffusion when the only data available are date and location of cases.

Association among Results of Serum ELISA, Faecal Culture and Nested PCR on Milk, Blood and Faeces for the Detection of Paratuberculosis in Dairy Cows

Paratuberculosis is a chronic, infectious disease of ruminants that entails a serious concern for the cattle industry. One of the main issues relates to the efficiency of diagnosis of subclinically infected animals. The objective of this field study was to analyse the association among results of a serum enzyme-linked immunosorbent assays (ELISA), faecal culture and nested PCR tests on milk, blood and faeces for Mycobacterium avium subsp. paratuberculosis detection in dairy cows. Faeces, blood and milk samples were collected from 328 lactating dairy cows in four known infected herds. Results were analysed to determine associations and levels of agreement between pairs of tests. A total of 61 animals (18.6%) tested positive when all the tests were interpreted in parallel. The agreement between results in different pairs of tests was poor, slight and fair in two, five and three of the 10 possible combinations respectively. Faecal culture and faecal polymerase chain reaction (PCR) resulted in the highest kappa coefficient (0.39; fair agreement), with the lowest agreement being for ELISA and blood PCR (-0.036; poor agreement). Fisher's exact test resulted in statistically significant associations (P < or = 0.05) between the following test pairs: ELISA : faecal culture; ELISA : faecal PCR; milk PCR : faecal PCR, blood PCR : faecal PCR and faecal culture : faecal PCR. Enzyme-linked immunosorbent assays showed the highest complementary sensitivity values for all the possible two-test combinations, followed by faecal PCR. The combined use of ELISA and faecal PCR has the potential to increase the overall sensitivity for the diagnosis of paratuberculosis infection.

Brucellosis of ruminants in Bosnia and Herzegovina: disease status, past experiences and initiation of a new surveillance strategy.

The current animal health situation in Bosnia and Herzegovina requires the prioritization of diseases for the application of control measures. One of the diseases requiring high priority is brucellosis of ruminants. Brucellosis is a zoonotic infectious disease and one of the most important zoonoses in the world. Brucellosis has been recognized during the past five decades as an important infectious disease in ruminants in Bosnia and Herzegovina. Control and eradication of brucellosis in animals is based on test and slaughter control policy. When the existing brucellosis control program was instituted, the veterinary and animal production sector was almost exclusively owned by the government, an arrangement that promoted compliance with the program and resulted in the successful control of the disease. This paper provides an overview of the current institutional and legislative framework for brucellosis control including the laboratory detection system and the epidemiological status of brucellosis in ruminants in Bosnia and Herzegovina. Relevant data were collected during the period spanning from the beginning of 2001 until the middle of 2007. Data we collected reveal an increase in the number of reported outbreaks in ruminants as well as a related increase in the number of human cases. This has brought serious consequences to public health, animal health and production and international trade.

Genetic Resistance to Johne's Disease in Four Cattle Breeds

Paratuberculosis is a chronic debilitating, infectious disease of ruminants, caused by Mycobacterium avium subsp. paratuberculosis, and characterized by progressive weight loss and profuse diarrhea. The disease has been suspected to have a genetic component and estimations of heritability of 0.15 have been reported. Crohn's disease (CD) is an inflammatory bowel disease in humans resembling many aspects of paratuberculosis. Susceptibility to CD has a strong genetic component and one gene involved has been identified as CARD15 (Caspase recruitment domain 15), with a role in the intracellular recognition of bacterial cell walls as a part of the immune response. The association between mutations in this gene and increased risk of CD has been extensively described. Since Johne's disease has been demonstrated to have a genetic component and genetic variation for infection resistance exists in cattle, CARD15 could also determine differences in resistance to paratuberculosis in the bovine. This candidate gene study focused on determining the alleles present in the CARD15 gene in a population of infected and of noninfected cows. Our objective was characterizing the distribution of two polymorphisms in this gene and testing their association with paratuberculosis infection in Florida cattle with the aim of finding resistance alleles useful in livestock selection.

Prediction of areas around the Mediterranean at risk of bluetongue by modelling the distribution of its vector using satellite imaging

Bluetongue is an infectious disease of ruminants caused by a virus transmitted by biting midges, one species of which, Culicoides imicola, is the major vector in the Old World. Following...

Serosurvey for Selected Disease Agents in White-tailed Deer from Mexico

Serum samples from 350 white-tailed deer (Odocoileus virginianus texanus) collected in March 1994 from northeastern Mexico were tested for the prevalence of antibody activity against five infectious diseases of ruminants. The prevalence rate was 81% for bluetongue virus (BTV) of all serotypes, 72% for epizootic hemorrhagic disease virus (EHDV), 3% for Borrelia burgdorferi, 69% for Anaplasma marginale, and 0% for Brucella abortus, B. melitensis, and B. ovis. These are diseases that affect domestic ruminants, and deer may act as a reservoir of infection. In addition, if deer are translocated, they may introduce pathogens to formerly disease-free areas. The high seroprevalence of BTV and EHDV cannot be related to the presence of hemorrhagic disease in the deer in this region. This is the first report to indicate the presence of B. burgdorferi infection of deer in Mexico. Despite the high prevalence of A. marginale titers, it is uncertain that deer play a role in the epizootiology of cattle anaplasmosis in the region. Apparently, white-tailed deer are unimportant in the epizootiology of brucellosis of both cattle and goats in northeastern Mexico.

Current world status of bluetongue

Bluetongue (BT) is an infectious disease of ruminants caused by viruses of the genus orbivirus. Currently there are 24 known serotypes of bluetongue virus worldwide. These viruses have been recognized on six continents, Africa, North America, South America, Australia, Asia and Europe, and currently five continents (all but Europe) have infection. South Africa has the greatest number of serotypes, followed by Australia and the United States.&#x0D; BTV is a double stranded RNA virus. There are 10 strands of RNA which code for the 7 structural and 3 nonstructural proteins. Gene segments 2 and 5 code for viral proteins 2 and 5. These proteins make up the outer coat and VP2 is responsible for neutralizing antigens. VP3 and 7 are the major proteins that make up the core. The core proteins, particularly VP7, is important for bluetongue group specificity.