Novel method for detecting bovine immunoglobulin G in dried porcine plasma as an indicator of bovine plasma contamination.

Current U.S. Food and Drug Administration regulations prohibit feeding of protein derived from mammalian tissue, excluding blood and blood products and any product that consists entirely of porcine or equine protein. A novel lateral flow immunoassay device has been developed that can quickly and qualitatively determine the presence of bovine immunoglobulin G (IgG), a major component in blood products, at very low levels (0.01% v/v). The device can be used to test for bovine IgG commingling in spray-dried porcine plasma used in the feed industry. Producers and consumers alike could use this device to verify product content at threshold levels.

Isolation of immunogenic outer membrane proteins from Mannheimia haemolytica serotype 1 by use of selective extraction and immunoaffinity chromatography.

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 iron-deficient 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 microg 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: lmmunogenic 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.

Evaluation of a lateral-flow immunoassay for use in monitoring passive transfer of immunoglobulins in calves.

OBJECTIVE: To determine the predictive ability of a commercially available lateral-flow immunoassay used for determining passive transfer of immunoglobulins in calves. ANIMALS: 204 male Holstein calves ranging from 4 to 8 days old. PROCEDURE: Serum samples were obtained from each calf. Results of refractometry, zinc sulfate turbidity technique, and the lateral-flow immunoassay were determined. Sensitivity, specificity, accuracy, and predictive ability were calculated on the basis of IgG concentrations determined by turbidimetric immunoassay (TIA). RESULTS: Mean IgG concentration in the study was 10.9 mg/ml as determined by TIA. Rate of failure of passive transfer in this study population was 56%. Associations between the values for the refractometry and zinc sulfate turbidity techniques were established by regression analysis. Accuracy for the lateral-flow immunoassay, refractometry, and zinc sulfate turbidity methods was 95, 80, and 73%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The lateral-flow immunoassay was better at determining the status of passive transfer of immunoglobulins, compared with the refractometry or zinc sulfate turbidity methods. The ability of the lateral-flow immunoassay to provide accurate results should enable clinicians to make immediate management or intervention decisions.

Rapid development of an efficacious swine vaccine for novel H1N1.

Recombinant hemagglutinin (HA) from a novel H1N1 influenza strain was produced using an alphavirus replicon expression system. The recombinant HA vaccine was produced more rapidly than traditional vaccines, and was evaluated as a swine vaccine candidate at different doses in a challenge model utilizing the homologous influenza A/California/04/2009 (H1N1) strain. Vaccinated animals showed significantly higher specific antibody response, reduced lung lesions and viral shedding, and higher average daily gain when compared to non-vaccinated control animals. These data demonstrate that the swine vaccine candidate was efficacious at all of the evaluated doses.

Isolation and characterization of the P5 adhesin protein of Haemophilus parasuis serotype 5.

Haemophilus parasuis is a Gram-negative respiratory pathogen of young pigs that colonizes the upper respiratory tract and produces a number of symptoms collectiviely described as Glässer's disease. Recently, an H. parasuis P5-like outer membrane adhesin protein homologous to H. influenzae P5 was identified. The P5 adhesin was partially purified by anion exchange and size-exclusion chromatography. Final purification for functional studies was performed by elution of the protein from a polyacrylamide gel. Identification of the protein as a P5 adhesin homolog of H. influenzae was confirmed by N-terminal sequencing. The P5 protein had a molecular mass of 32,000 and a pI of 5.5. Unlike the H. influenzae P5 adhesin, the H. parasuis P5 protein did not bind carcinoembryonic antigen.