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Haemophilus parasuis is an important swine pathogen that causes Glasser's disease, characterized by pneumonia, polyserositis and meningitis. Protection against H. parasuis infection is associated with the presence of homologous antibodies in serum. However, a H. parasuis antigen that can elicit a protective immune response against all H. parasuis strains has yet to be found. A novel immunogenic and species-specific H. parasuis protein was identified by screening H. parasuis whole cell proteins using swine convalescent sera. One protein of 52kDa was clearly immunodominant and conserved among different H. parasuis strains. This protein was further identified as an oligopeptide permease A (OppA). Because OppA elicited a specific antibody response in pigs that recovered from H. parasuis infection, we investigated its potential role in diagnostics and protective immunity. An ELISA test using recombinant OppA (rOppA) as its coating antigen was further developed and tested. H. parasuis specific antibodies to rOppA were detected in serum from convalescent pigs but not in serum from specific pathogen free (SPF) or conventional pigs. Pigs immunized with rOppA protein had robust serological responses. However, the antibodies were not protective against challenge infection. We conclude that OppA is a universal species-specific H. parasuis immunogen, and a good marker for previous systemic infection with H. parasuis., (Copyright © 2016. Published by Elsevier Ltd.)

Porcine deltacoronavirus (PDCoV) was identified in multiple states across the United States (US) in 2014. In this study, we investigate the presence of PDCoV in diagnostic samples, which were further categorized by case identification (ID), and the association between occurrence, age, specimen and location between March and September 2014. Approximately, 7% of the case IDs submitted from the US were positive for PDCoV. Specimens were categorized into eight groups, and the univariate analysis indicated that oral fluids had 1.89 times higher odds of detecting PDCoV compared to feces. While the 43-56 day age group had the highest percentage of PDCoV positives (8.4%), the univariate analysis indicated no significant differences between age groups. However, multivariable analysis for age adjusted by specimen indicated the >147 day age group had 59% lower odds than suckling pigs of being positive for PDCoV. The percentage of PDCoV in diagnostic samples decreased to <1% in September 2014. In addition, 19 complete PDCoV genomes were sequenced, and Bayesian analysis was conducted to estimate the emergence of the US clade. The evolutionary rate of the PDCoV genome is estimated to be 3.8×10(-4) substitutions/site/year (2.3×10(-4)-5.4×10(-4), 95% HPD). Our results indicate that oral fluids continue to be a valuable specimen to monitor swineherd health, and PDCoV has been circulating in the US prior to 2014., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)

Porcine deltacoronavirus (PDCoV) was first reported in the United States (US) in February 2014. This was the second novel swine enteric coronavirus detected in the US since May 2013. In this study, we conducted retrospective testing of samples submitted to three veterinary diagnostic laboratories where qualifying biological samples were derived from previously submitted diagnostic case submissions from US commercial swine farms with a clinical history of enteric disease or from cases that had been previously tested for transmissible gastroenteritis virus, PEDV, or rotavirus. Overall, 2286 banked samples were tested from 27 States. Samples were collected in 3 separate years and in 17 different months. Test results revealed 4 positive samples, 3 collected in August 2013 and 1 collected in October 2013. In addition, a case series including 42 operations in 10 States was conducted through administration of a survey. Survey data collected included information on characteristics of swine operations that had experienced PDCoV clinical signs. Special emphasis was placed on obtaining descriptive estimates of biosecurity practices and disease status over time of each operation. Clinical signs of PDCoV were reported to be similar to those of PEDV. The average number of animals on each operation exhibiting clinical signs (morbidity) and the average number of case fatalities was greatest for suckling and weaned pigs. Average operation-level weaned pig morbidity was greatest in the first week of the outbreak while average operation-level suckling pig case fatality was greatest in the second week of the outbreak. The survey included questions regarding biosecurity practices for visitors and operation employees; trucks, equipment and drivers; and feed sources. These questions attempted to identify a likely pathway of introduction of PDCoV onto the operations surveyed., (Published by Elsevier B.V.)

In early 2014, a Minnesota sow farm with a solid vaccination history suffered a severe porcine reproductive and respiratory syndrome (PRRS) outbreak with unusually high morbidity and mortality in piglets and sows, as well as anorexia and secondary bacterial infections in nursery pigs. Due to the unusual clinical severity in a PRRS-immune herd, genetic characteristics of the virus were examined to determine if a new PRRSV genotype had emerged. Phylogenetic analysis indicated that the virulent strain (PRRSV2/USA/Minnesota414/2014) was related to virulent strains circulating in the mid-western United States in recent years, and that the nonstructural protein 2 (nsp2) gene of MN414 contained an insertion-deletion pattern typical of existing type 2 virulent strains. We conclude that the MN414 isolate is a recently evolved member of the virulent lineage 1 family of type 2 PRRSV., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Metabolic surgery can modulate weight as well as food intake and basal energy expenditure. In this study, we evaluate the effectiveness of duodenal exclusion by analysing anthropometric results, intake variations, food behaviour and calorimetric parameters., Methods: This is an experimental study with 8-week-old Sprague-Dawley male rats. The sequences used are as follows: Cafeteria diet for 3 weeks, followed by surgery and sacrifice at 4 weeks. Four experimental groups are as follows: two non-obese groups (n = 15; surgery = 10, sham = 5) and two obese groups by cafeteria diet (n = 15; surgery = 10, sham = 5). Surgery performed was duodenal exclusion with physical barrier. Weight, intake, glycaemia and basal energy expenditure by indirect calorimetry were monitored before and after surgery., Results: Weight changes in groups that underwent intervention were significant. The reduction in calorie consumption after surgery was significant in the obese intervention group despite an increased standard feed consumption (161 ± 11 vs 139 ± 13 Kcal/day, p < 0.05; due to a lower consumption of cafeteria diet). In non-obese animals, changes were transient. Basal energy expenditure decreased in both intervention groups: 6.2 ± 0.5 vs 5.5 ± 0.4 Kcal/kg/h in non-obese animals and 5.6 ± 0.3 vs 4.7 ± 0.3 Kcal/kg/h in obese animals (p < 0.05)., Conclusions: Duodeno-jejunal tube placement stops weight gain in obese and non-obese animals. In obese animals, there is an important qualitative change in appetite towards standard feed with a significant decrease in caloric intake. In non-obese animals, changes in quantitative intake are transient. This surgery decreases basal energy expenditure in obese animals. This may be attributed to an enhanced thermogenic effect of food and a slowing in the animal's weight gain.

Porcine epidemic diarrhea virus (PEDV), which emerged in the United States in 2013, has spread throughout North America. Limited availability of PEDV complete genomes worldwide has impeded our understanding of PEDV introduction into the United States. To determine the relationship between the North American strains and global emerging and historic PEDV strains, we sequenced and analyzed complete genomes of 74 strains from North America; the strains clustered into 2 distinct clades. Compared with the initially reported virulent US PEDV strains, 7 (9.7%) strains from 4 states contained insertions and deletions in the spike gene (S INDELs). These S INDEL strains share 99.8%-100% nt identity with each other and 96.2%-96.7% nt identity with the initial US strains. Furthermore, the S INDEL strains form a distinct cluster within North American clade II, sharing 98.6%-100% nt identity overall. In the United States, the S INDEL and original PEDV strains are co-circulating and could have been introduced simultaneously.

In February 2014, porcine deltacoronavirus (PDCoV) was identified in the United States. We developed a PDCoV real-time reverse transcription PCR that identified PDCoV in 30% of samples tested. Four additional PDCoV genomes from the United States were sequenced; these had ≈99%-100% nt similarity to the other US PDCoV strains.

Surveillance of porcine reproductive and respiratory syndrome (PRRS) in negative sow farms is usually performed by testing for the presence of antibodies against PRRS virus in serum with a commercial ELISA test. The objective of this study was to evaluate the feasibility of pooling serum samples for detection of PRRS virus antibodies by ELISA. The effect of pool size on the sensitivity and specificity of the ELISA test was evaluated by testing true positive samples and false positive samples, respectively, diluted in negative sera. The effect of three different cut-off values for the interpretation of the diagnostic test (0.4, 0.3 and 0.2) was evaluated as well. Furthermore, the obtained sensitivity and specificity estimates were used to calculate the herd sensitivity and herd specificity of surveillance protocols in different scenarios. The results showed that pooling serum samples to detect PRRSV antibodies resulted in a decrease in sensitivity and an increase in specificity, compared to testing individual samples, while the reduction of the s/p cut-off value recommended by the manufacturer (0.4) had the opposite effect. We describe an approach that can increase the herd sensitivity of a surveillance protocol for breeding herds, while maintaining high herd specificity and low testing costs. This can be achieved by sampling a larger number of animals and running the samples in pools. Therefore, the conventional monitoring protocols based on ELISA on individual samples can be improved by using pooled-sample testing.

Background: Ghrelin is an important factor in the regulation of intake. Most ghrelin is synthesized in the gastric fundus, but this is not the only location. The aim of this experimental study was to analyze the effect of sleeve gastrectomy (removing fundus) on the volume of intake in four experimental models and determine how this relates to changes in weight, plasmatic levels of glycemia, ghrelin, GLP-1, and insulin., Methods: Sleeve gastrectomy was performed on four experimental models: (1) non-obesity; (2) exogenous obesity caused by excessive calorie intake; (3) genetically determined obesity (Zucker rats); and (4) genetically determined obesity and type 2 diabetes mellitus (Zucker diabetic fatty; ZDF rats). Model 2 had a control group on which sleeve gastrectomy was not performed., Results: In the non-obese group, there were few changes after intervention, but in model 2, sleeve gastrectomy led to normalization of weight and endocrine-metabolic parameters that were the same as those for non-obese rats. The exception was for GLP-1, which has an anorexigenic effect: GLP-1 remained higher. In Zucker rats, sleeve gastrectomy had a slight effect on all parameters. In ZDF rats, sleeve gastrectomy led to a reduction in intake and a stabilization of weight., Conclusions: Sleeve gastrectomy is a very good option for exogenous obesity. Normalization of hormonal levels led us to find an extragastric ghrelin production.

Porcine circovirus type 2 (PCV2) is detected in high amounts within the characteristic microscopic lesions of postweaning multisystemic wasting syndrome (PMWS) affected pigs. In spite of recent advances on disease pathogenesis, the precise cell types that support viral replication are still a major issue of scientific discussion. In this study, a new methodology to detect cell types that support PCV2 replication was designed. For this purpose, two in situ hybridisation (ISH) methods were developed and applied on tissues of PMWS naturally affected pigs using two probes designed from the ORF1 sequence of the virus. While the complementary probe (CP) detected ssDNA, ORF1 mRNA and replicative form (RF) of PCV2, the RF probe (RFP) exclusively hybridised with the RF of the virus, thus, only labelling cells where PCV2 replication is taking place. Both probes demonstrated to be specific and equally sensitive by an in vitro Southern blot hybridisation assay. ISH labelling with the CP was extensive in lymphoid tissues and of variable amount in other non-lymphoid tissues. With this probe, mainly macrophage-like cells were labelled but also other cell types such as hepatocytes and other epithelial cells. Tissues in which RFP labelling was found more frequently were lung, inguinal and mesenteric lymph nodes, tonsil and liver. Labelling with the RFP was always nuclear, and found in the same cell types as with the CP, although in a relatively low proportion of them; labelling of macrophage-like cells was infrequent. Therefore, the results indicate that at least a certain proportion of macrophages may support PCV2 replication, but main cells where PCV2 replicates are of epithelial/endothelial origin. In summary, the present study permitted the study of cell types that support PCV2 replication by the use of ISH on formalin-fixed, paraffin-embedded tissues of PMWS affected pigs.