The ability of Mycoplasma hyopneumoniae to agglutinate RBC was evaluated to develop an in vitro cytadsorption assay. Using swine RBC in a microtitration hemagglutination test, no agglutination or partial agglutination was detected. Comparison of RBC from various other species indicated that improved hemagglutination was obtained with RBC from turkeys. This hemagglutination was detected only when mycoplasma cells used in the assay had been frozen and thawed, heated at 50 C for 30 minutes, or treated with trypsin. Treatment of RBC with trypsin or neuraminidase enhanced hemagglutination. Possible surface lectin activity in M hyopneumoniae was evaluated by use of carbohydrates in a blocking assay; hemagglutination was not inhibited by any of 13 carbohydrates evaluated. Mycoplasma hyopneumoniae convalescent porcine serum and monoclonal antibodies against 2 M hyopneumoniae immunogens of molecular weights of 64,000 and 41,000 inhibited hemagglutination.

Snail eggs can be the raw material for the production of a caviar substitute. The substitute varies from the original in caloric value and nutrient content which determine the nutritional value of every foodstuff. The present study aimed to determine and compare the nutritional value and protein quality of eggs from two subspecies of edible snail. The chemical composition of the snail eggs i.e. Cornu aspersum maxima and Cornu aspersum aspersum was determined in accordance with international standards. In order to evaluate the protein quality of the eggs of the two studied snail subspecies, the chemical score (CS), and a reference protein were used. Significant differences in the content of water, ash, and carbohydrates, but comparable protein and fat contents and caloric values were found. The protein in the eggs of the snails was complete by the measure of the model adopted for this study, however, meeting the daily essential amino acid requirements of an adult would require an immense supply of both species’ eggs. Snail eggs of the Cornu genus were characterised by much lower nutritional value in comparison with caviar and caviar substitutes.

Sheep pulmonary adenomatosis (ovine pulmonary adenomatosis, OPA, Jaagsiekte) is a chronic contagious bronchoalveolar carcinoma caused by the Jaagsiekte sheep retrovirus. Since effective treatment and a vaccination procedure are not currently possible, control and eradication of the disease is difficult. It leads to serious economic losses around the world, therefore studies are currently underway in order to design control and eradication programmes. In this study, levels and changes in selected tumour markers (carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 125, CA 19-9, CA 15-3, and alphafetoprotein (AFP)-3) and their diagnostic significance were investigated. A total of 30 sheep were used. Clinical examinations were performed and blood samples were obtained before slaughter from all animals with presumed OPA. Blood samples with positive OPA results by macroscopic and histopathological examination were included in the study as the experimental group and numbered 20. Sheep totalling 10 had negative OPA results and provided control samples. CEA levels were similar in both groups, and the differences were statistically insignificant (P > 0.05). CA 125, CA 19-9, CA 15-3, and AFP-3 levels were higher in the OPA group than the control group and with statistical significance (P < 0.05). In all OPA animals, CA 125 levels were higher than 1 U/mL. serum CAs and AFP levels increase significantly in adenomatous sheep. These tumour markers are thought to facilitate the diagnosis of OPA.

Four healthy calves were inoculated with Pasteurella haemolytica serotype 1 by instillation of a broth culture into the middle nasal meatus of the left nostril. Four weeks later, calves were exposed to infectious bovine rhinotracheitis virus by aerosol into both nostrils. All calves became ill, from approximately day 3 through day 10 after virus exposure, and shed increased amounts of nasal mucus. Two calves were induced to shed P haemolytica by the virus infection, and 2 calves required reinoculation with P haemolytica for nasal passages to become actively colonized. Elastase activity in nasal mucus increased about 15-fold within 3 days and peaked about 60-fold over baseline by 7 days after virus exposure. Activity of N-acetyl-beta-D-glucosaminidase, a measure of cell damage and serum leakage, increased slightly by day 3 and reached plateau on day 5, almost threefold over baseline activity. Protein and carbohydrate content increased at a rate similar to that of N-acetyl-beta-D-glucosaminidase activity with about 12-fold and sixfold increases, respectively. None of the variables returned to baseline by 19 days after virus exposure, Increased elastase activity preceded colonization by P haemolytica and decreasing elastase activity preceded decreasing P haemolytica concentration in the nasal secretions. A causal relation between elastase activity and P haemolytica colonization could be mediated by cleavage of epithelial cell surface fibronectin and exposure of receptors.

The balance of coagulation and fibrinolysis was studied in 15 horses during the prodromal stages of acute laminitis induced by carbohydrate overload. Progression of the disease was stopped 12 to 24 hours before the expected onset of lameness in trial 1 (8 horses) and at the onset of lameness in trial 2 (7 horses). The end points in each trial were identified by specific changes in blood pressures (trial 1) and by changes in pulse, rectal temperature, and arterial pressure (trial 2) that were anticipated on the basis of original description of the experimental model. Blood samples for hemostasis evaluation were collected before and after carbohydrate overload in trial 1 and after carbohydrate overload in trial 2. Significant changes were not detected in platelet count, mean platelet volume, prothrombin time, activated partial thromboplastin time, fibrinogen concentration, plasminogen concentration, alpha-2-antiplasmin, antithrombin III, protein C, thromboxane B2, or fibrin(ogen) degradation product concentration. We concluded that an imbalance in coagulation and fibrinolysis is not pathogenic in the onset of experimentally induced equine acute laminitis. Because several test methods used to evaluate hemostasis in these horses were new, reference values for 34 healthy adult horses were established.

The antibody responses to the capsular carbohydrate (CC) purified from Pasteurella haemolytica serotype 1 were determined by an ELISA using 135 sera from 6 calves vaccinated with phosphate-buffered saline solution, formalin-killed P haemolytica bacterins, live P haemolytica, or an extract of P haemolytica referred to as carbohydrate-protein subunit (CPS). Calves vaccinated with live P haemolytica, bacterins, or CPS developed serum antibodies to CC. Bacterins containing Freund incomplete adjuvant or Freund complete adjuvant induced higher antibody responses than did bacterins containing aluminum hydroxide. In 4 of 6 experiments, high antibody responses to CC were significantly (P less than 0.05) correlated with resistance to transthoracic challenge exposure with P haemolytica. When calves were challenge exposed with a dose of P haemolytica that was 4.5 times greater than the standard challenge exposure dose or when calves that had been vaccinated with CPS were challenge exposed, antibody responses did not significantly (P greater than 0.05) correlate with resistance to challenge exposure. The amount of serum antibodies to CPS increased significantly (P less than 0.05) when calves were vaccinated with live or killed P haemolytica or with CPS, compared with that in calves given saline solution. In 5 of 6 experiments, correlation between high antibody responses and resistance to challenge exposure was significant (P less than 0.05). The correlation between those variables was not significant (P less than 0.07) for CPS-vaccinated calves. In the ELISA, treatment of CPS with sodium m-periodate, to oxidize periodate-sensitive carbohydrate epitopes, failed to markedly alter the antibody response to CPS. However, the correlation between high antibody responses to periodate-treated CPS and resistance was significant (P less than 0.05) for all 6 experiments. In the ELISA, periodate treatment of CC, lipopolysaccharide, and CPS caused average reductions in antibody reactivity of 7.1%, 53.8%, and 34.5%, respectively. Preadsorption of sera with CC or lipopolysaccharide did not markedly reduce antibody reactivity with CPS. Preadsorption of sera with CC and reaction with periodate-treated and nontreated CPS indicatedthat for calves given phosphate-buffered saline solution vaccines, antibody reactivity was reduced 65.4%, whereas for those vaccinated with a bacterin with aluminum hydroxide, a bacterin with Freund incomplete adjuvant, or live P haemolytica, antibody reactivity was reduced 47.1%, 40.5%, and 25.0%, respectively. It was concluded that serum antibodies to CC are of some importance in resistance and that certain epitopes in CPS that are not sensitive to periodate are of importance in resistance to bovine pneumonic pasteurellosis. There are qualitative and quantitative differences among the serum antibody responses to carbohydrate epitopes for calves vaccinated with phosphate-buffered saline solution, bacterins, or live P haemolytica.

The ability of Mycoplasma hyopneumoniae to agglutinate RBC was evaluated to develop an in vitro cytadsorption assay. Using swine RBC in a microtitration hemagglutination test, no agglutination or partial agglutination was detected. Comparison of RBC from various other species indicated that improved hemagglutination was obtained with RBC from turkeys. This hemagglutination was detected only when mycoplasma cells used in the assay had been frozen and thawed, heated at 50 C for 30 minutes, or treated with trypsin. Treatment of RBC with trypsin or neuraminidase enhanced hemagglutination. Possible surface lectin activity in M hyopneumoniae was evaluated by use of carbohydrates in a blocking assay; hemagglutination was not inhibited by any of 13 carbohydrates evaluated. Mycoplasma hyopneumoniae convalescent porcine serum and monoclonal antibodies against 2 M hyopneumoniae immunogens of molecular weights of 64,000 and 41,000 inhibited hemagglutination.

OBJECTIVE To identify cardiac tissue genes and gene pathways differentially expressed between dogs with and without dilated cardiomyopathy (DCM). ANIMALS 8 dogs with and 5 dogs without DCM. PROCEDURES Following euthanasia, samples of left ventricular myocardium were collected from each dog. Total RNA was extracted from tissue samples, and RNA sequencing was performed on each sample. Samples from dogs with and without DCM were grouped to identify genes that were differentially regulated between the 2 populations. Overrepresentation analysis was performed on upregulated and downregulated gene sets to identify altered molecular pathways in dogs with DCM. RESULTS Genes involved in cellular energy metabolism, especially metabolism of carbohydrates and fats, were significantly downregulated in dogs with DCM. Expression of cardiac structural proteins was also altered in affected dogs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that RNA sequencing may provide important insights into the pathogenesis of DCM in dogs and highlight pathways that should be explored to identify causative mutations and develop novel therapeutic interventions.

Objective-To use antibodies produced by calves in response to infection with Mannheimia haemolytica in immunoaffinity chromatography for the identification and subsequent isolation of the dominant immunogenic antigens from bacteria grown in irondeficient media. Sample Population-Serum from 10 calves actively infected with M haemolytica. Procedure-An outer membrane protein fraction was obtained from sonicated salt-extracted M haemolytica cells by extraction with N-lauroyl sarcosinate. The immunoglobulin fraction of serum from calves actively infected with M haemolytica was used to prepare an immunoaffinity column. The immunoaffinity column was used to isolate the dominant immunogenic proteins from the outer membrane protein fraction. The resultant immunogenic protein fraction was subjected to ELISA and immunoblot methods as well as carbohydrate quantification. Sequencing of the N-terminal was performed on the most prominent protein. Results-5 immunogenic proteins with molecular weights of 42, 30, 24, 20, and 15 kd were isolated. The immunogenic protein fraction was found to contain 51% carbohydrate. The immunoaffinity column capacity was 1 µg of immunogenic protein/mL of gel. The N-terminal sequence of the 42-kd protein was Tyr-Gln-Thr-Tyr-Gln-Ser-X-Leu-Gln, where X could not be identified. Conclusions and Clinical Relevance-Immunogenic proteins were isolated by use of immunoaffinity chromatography. A substantial amount of carbohydrates was co-purified in the process. Additional experiments are needed to determine whether the carbohydrates would hinder or enhance development of vaccine preparations. This method could potentially allow a more rapid production of antigens for use in vaccines.

Bordetella avium is an important respiratory tract pathogen of turkeys. In common with other pathogenic bordetellae, B avium manifests a tissue tropism for cilia of the respiratory tract epithelium. To determine the molecular characteristics of the host cell receptors for B avium, we used hemagglutination and in vivo adherence assays. Carbohydrates, mucus, sialic acid-specific lectin, and other glycoconjugates were evaluated for their ability to competitively inhibit binding of B avium to host cells. The gangliosides, GD(1a) and GT(1b), completely inhibited hemagglutination, whereas N-acetylneuraminic acid (sialic acid) partially inhibited hemagglutination. Adherence to turkey tracheal mucosa in vivo was significantly (P < 0.01) inhibited by GD(1a) and GT(1b) gangliosides, N-acetylneuraminic acid, bovine sub-maxillary mucin, and horseshoe crab (Limulus polyphemus) lectin. Treatment of the tracheal mucosa with neuraminidase also inhibited adherence of B avium. We conclude that N-acetylneuraminic acid and the gangliosides, GD(1a) and GT(1b) may be important components of the tracheal mucosa receptor for B avium in turkeys.