Immunization of dogs against Echinococcus granulosus using worm secretory antigens.

A new approach to the immunization of dogs against Echinococcus granulosus, using secretory antigens derived from adult tapeworms cultivated in vitro, resulted in a highly significant suppression of egg production by Echinococcus granulosus in immunized dogs. There was also a significant reduction in the number of proglottids per worm in immunized dogs. It appeared that the suppression of egg production was due to an immune effect on both somatic growth and germinal development. The implications of this work with respect to hydatidosis control programmes are discussed.     

Studies on glutamic-oxalacetic (GOT) and glutamic-pyruvic (GPT) transaminases of swine kidney worm Stephanurus dentatus (Diesing, 1839). I. Assay and general properties.

Biochemical studies on the two transaminases GOT and GPT of swine kidney worm Stephanurus dentatus have been made. GOT has been found much more active than GPT. Enzyme activities are based on the formation of oxaloacetate (GOT) or pyruvate (GPT) from aspartic acid and alanine respectively with oxoglutarate. A linear relationship is observed between the enzyme concentration and activity. GOT shows a maximum activity at pH 8.0 and Michaelis constant 9 X 10(-3) M for male and 2.9 X 10(-3) M for female. GPT has an optimum pH of 7.5 and a Michaelis constant 19 X 10(-3) M for male and 8 X 10(-3) M for female. The optimum temperature for both GOT and GPT was 60 degrees C.     

Transplantation in miniature swine. XII. N-terminal sequences of class I histocompatibility antigens (SLA) and beta 2-microglobulin

Purified class I histocompatibility antigens (SLA) from three haplotypes were prepared by papain treatment of lymphoid cell membranes obtained from spleens and lymph nodes of miniature swine homozygous at their major histocompatibility complex. Antigens were purified by ion-exchange chromatography followed by gel filtration. Purity was analyzed by SDS-PAGE, and antigenic specificity by inhibition of complement-dependent, alloantiserum-mediated cytotoxicity. The SLA antigens were reduced and alkylated, and the component heavy and light chains were isolated by gel filtration under dissociating conditions. N-terminal amino acid sequences were obtained for SLAaa, SLAcc, and SLAdd heavy chains, as well as for the light chain, beta 2-microglobulin. The swine antigens showed high levels of homology with class I antigens from other animal species. Heterogeneity was observed among the swine haplotypes, and several of the positions at which substitutions were found are apparently invariant in other animal species. In contrast, only minimal sequence heterogeneity was detected within haplotypes, the basis of which may be of relevance to understanding the evolutionary development of these molecules.     

Biochemical and histochemical studies on non-specific phosphomonoesterases of swine kidney worm Stephanurus dentatus (Diesing, 1839).

Biochemical and histochemical studies have been made on non-specific acid and alkaline phosphomonoesterases of S. dentatus. The two forms of acid phosphomonoesterases have been found active at pH 4.0 and 6.0. The pH optima for the two forms of aklaline phosphomonoesterases lie at 8.0 and 10.0. Studies on the distribution of acid and alkaline phosphomonoesterases in various tissues have revealed an abundance of acid phosphomonoesterase in various parts of the alimentary canal and various organs of the reproductive system. The excretory ducts show alkaline phosphomonoesterase activity only.     

Crystal formation of swine vesicular disease virus in porcine kidney cells (IB-RS-2 cells)

Crystal formation of swine vesicular disease virus (SVDV) in IB-RS-2 cells was studied by electron microscopy. Cells were harvested 0, 3, 3.5, 4, 4.5, 5, 6 and 7 hours after inoculation. Crystalline arrays of SVDV was first observed in the cytoplasm of a few cells 4.5 hours after inoculation. In the cytoplasm of many cells harvested at 5 hours, 1 to 3 crystalline arrays of SVDV were observed. After that, a small number of cells had crystalline arrays in the cytoplasm. The cells with crystalline arrays were rich in ribosome and polysome with dilated mitochondria and many tiny vesicles. An individual virus particle was ca. 18 nm in diameter, and the center-to-center space ca. 22 nm. Crystalline arrays varied in size depending on the plane of section.     

Spread of porcine circovirus associated disease (PCVAD) in Ontario (Canada) swine herds: Part I. Exploratory spatial analysis

Background  

The emergence of porcine circovirus associated disease (PCVAD) was associated with high mortality in swine populations worldwide. Studies performed in different regions identified spatial, temporal, and spatio-temporal trends as factors contributing to patterns of the disease spread. Patterns consistent with spatial trend and spatio-temporal clustering were already identified in this dataset. On the basis of these results, we have further investigated the nature of local spread in this report. The primary objective of this study was to evaluate risk factors for incidence cases of reported PCVAD.     

Lesions in mink (Mustela vison) infected with giant kidney worm (Dioctophyma renale).

Adult Dioctophyma renale occupied the enlarged renal pelvis of the right kidney of naturally infected mink. Lesions in the kidney parenchyma consisted of connective tissue proliferation in the interstitial tissue, tubular atrophy and fibrosis, and periglomerular fibrosis. The luminal surface of the renal pelvis wall was formed of numerous papillae covered with transitional epithelium. The nematodes in the lumen were bathed in an albuminous fluid containing red blood cells, epithelial cells and D. renale eggs. The left (uninfected) kidney was 60% larger than the left kidney of normal mink.     

First report of the giant kidney worm (Dioctophyme renale) in a harbor seal (Phoca vitulina)

A male harbor seal (Phoca vitulina) was found moribund on the coast of New Jersey in January of 2003 and died a few hours later in the Marine Mammal Stranding Center. On necropsy, a single female Dioctophyme renale was recovered from the peritoneal cavity, and a tissue mass was found adjacent to the pelvic urethra and urinary bladder. Within this tissue mass were found D. renale ova. This is the first report of this nematode in the harbor seal and in a North American marine mammal.     

Somatic antigens of Shigella. The structure of the specific polysaccharide of Shigella newcastle (Sh. flexneri type 6) lipopolysaccharide.

The specific polysaccharide was obtained from the lipopolysaccharide of Shigella newcastle by mild acid hydrolysis and further purified by permeation chromatography on Sephadex G-50. It was found to consist of L-rhamnose, 2-acetamido-2-deoxy-D-galactose, D-galacturonic acid residues and O-acetyl groups in the molar ratios of 2:1:1:1. On the basis of 1H and 13C nuclear magnetic resonance spectroscopy, methylation analysis, partial acid hydrolysis, Smith degradation, and chromium trioxide oxidation, the following structure can be assigned to the repeating oligosaccharide unit of the polysaccharide:-4)DGalA(beta 1-3)DGalNAc-(beta 1-2)LAc3Rha(alpha 1-2)LRha(alpha 1-, where GalA = galacturonic acid. GalNAc = N-acetylgalactosamine, Ac3Rha = 3-O-acetylrhamnose. The structural and immunochemical data presented prove that Sh. newcastle lipopolysaccharide belongs to a 'non-classical' type of somatic antigens with acidic O-specific polysaccharide chains.     

Somatic antigens of Pseudomonas aeruginosa. The structure of the polysaccharide chain of Ps. aeruginosa O-serogroup 7 (Lanyi) lipopolysacharide

A loosely bound lipopolysaccharide-protein complex was extracted from cells of Pseudomonas aeruginosa strain 170015 (O:7ab; Lanyi classification) by saline solution and purified from contaminant nucleic acid by Cetavlon treatment followed by precipitation in an ultracentrifuge. The saline-treated cells were re-extracted with hot aqueous phenol to give firmly bound lipopolysaccharide which was isolated from the phenol layer and purified by ultracentrifugaiton. The identity of both lipopolysaccharide preparations was proved by serological and chemical evidence. Mild acid degradation of the lipopolysaccharide resulted in the splitting off of a lipid component and led to polysaccharide which was purified by gel-filtration on a Sephadex G-50 column. The polysaccharide consisted of N-acetyl-D-fucosamine, N-acetyl-L-fucosamine and D-glucose in the ratio 1:1:1. On the basis of nuclear magnetic resonance spectra, results of methylation analysis and two sequential Smith degradations, the following structure can be assigned to the repeating unit of the polysaccharide: -3)LFucNAc(alpha 1-3)DFucNAc(beta 1-2)DGlc(beta 1-. The polysaccharide did not show serological activity whereas alkali-treated lipopolysaccharide readily sensitised sheep erythrocytes and inhibited the passive haemagglutination reaction with anti-(O:7a,b)serum. Evidence is presented that the oligosaccharide repeating units of the polysaccharide and alkali-treated lipopolysaccharide are indistinguishable. Ps. aeruginosa strain 170016 (O:7a,c) was shown to have the O-specific lipopolysaccharide identical with that from strain 170015. The presented data show that subfactors 7b and 7c in the Lanyi classification of Ps. aeruginosa O-antigens seem to relate to components of the bacterial surface other than lipopolysaccharides.     

Heat-labile antigens of Salmonella enteritidis. I. Extraction of antigens

Milne, Margaret (University of Adelaide, Adelaide, Australia), and F. M. Collins. Heat-labile antigens of Salmonella enteritidis. I. Extraction of antigens. J. Bacteriol. 92:543-548. 1966.-Salmonella enteritidis strains of high and low mouse virulence were grown in continuous culture. Cell walls were obtained from both strains by sonic disruption, and the washed walls were extracted at 4 C with sodium dodecyl sulfate or sodium deoxycholate. The extracts were eluted with a saline gradient from a diethylaminoethyl cellulose column, and the separated peaks were tested for precipitin activity against rabbit antisera prepared with heator ethyl alcohol-killed vaccines. The separated antigens reacted less intensely with the antiserum to heat-killed cells than they did to the antisera prepared against alcohol-killed vaccine. Little antigenic difference could be detected between the extracts prepared from the virulent and the avirulent organisms.     

Somatic embryogenesis in cotton (Gossypium) I. Effects of source of explant and hormone regime

Optimal media for induction of somatic embryogenesis from mature and immature tissues ofG. hirsutum L. cv Coker 312 were determined. Explants of three-day-old seedlings form somatic embryos in 100% of cultures when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 0.5 mg/1 kinetin. Mature tissues are more recalcitrant than immature tissues and formed somatic embryos on a limited number of hormone treatments. Stem tissue is most readily induced to form somatic embryos by 2 mg/1 napthaleneacetic acid plus 0.1 mg/1 kinetin, whereas leaf tissue formed embryos best when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 1.0 mg/1 (2-isopentyl)-adenine, or 1.0 mg/1 napthaleneacetic acid plus 0.5 mg/1 (2-isopentyl)-adenine.     

Transplantation in miniature swine. V. Characterization of Ia antigens.

The complexity of the I region analog associated with the MHC of miniature swine has been probed by sequential antibody precipitation studies of Ia antigens. Treatment of solubilized lymphocyte preparations from MSLA homozygotes of the DD haplotype with excess AA anti-DD alloantiserum led to precipitation of only a portion of the total Ia antigens, as determined by secondary precipitation of the remaining material with CC anti-DD serum. The presumed I region of miniature swine must therefore code for more than one Ia antigen-bearing polypeptide chain. In addition, certain mouse alloantisera that had previously been shown to react with rat Ia antigens were tested for reactivity with swine Ia antigens. Anti-Iak mouse alloantisera precipitated Ia molecules from every swine extract tested, regardless of MHC type, precluding genetic mapping studies. However, sequential precipitation studies demonstrated that the cross-reactive mouse alloantisera reacted only with a subclass of swine Ia antigens, again suggesting genetic complexity of the pig Ia locus.     

Standardization of FIAXtm for schistosomiasis using crude cercarial and adult worm antigens

A fluoroimmunoassay (FIAXTM) has been developed for quantitating the antibody response to schistosomiasis by using cercarial and adult worm antigens of Schistosoma mansoni. The FIAXTM assay was calibrated by using an enzyme-linked immunosorbent assay (ELISA) performed with the same antigens. Studies of reproducibility and preliminary data on a battery of sera from proven cases of schistosomiasis an uninfected control sera are presented.     

Monoclonal antibodies against rat kidney antigens.

The proximal tubule of the nephron is subdivided into three structurally and functionally distinct segments, which can be differentiated with the help of special methods. With the aim of producing selective markers for these three portions of the proximal tubule, we raised monoclonal antibodies against the brush border membranes of the rat kidney. Immunohistochemistry was carried out with eleven different monoclonal antibodies to sections of rat kidney and other tissues at the light- and electron-microscopical level. These monoclonal antibodies mainly detect antigens located on the brush border of the proximal tubule, and they allow a distinction between its three segments. However, some antibodies also recognize other portions of the nephron, or even the glomerulus or stromal elements. Sites recognized by the antibodies are not limited to the kidney, but staining is observed on the intestinal brush border, the intralobular ducts of the pancreas, the bile canaliculi of the liver and on the macrophage clusters of the spleen. These antibodies are interesting reagents which can be applied to study biochemical differences between brush border membranes. In addition, they recognize antigenically related sites in other organs with reabsorptive or secretory tasks.     

Immunization with cellulose-immobilized antigens. The development of A. E. Gurvitch concept

A highly purified TUL4-CBD chimeric protein was obtained by one stage purification method. TUL4-CBD protein consists of TUL4 Francisella tularensis mature peptide sequence, Gly-Ser spacer and cellulose binding domain (CBD) of Anaerocellum thermophilum. The TUL4-CBD protein was shown to induce production of specific antibodies to TUL4 protein in laboratory animals.