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Binding of enteric bacteria to hog gastric mucin.

The binding features of enteric bacteria were studied using a model mucin of hog gastric origin. The time requirement of binding is short, it is temperature-independent, but dose-dependent. The binding effectiveness of Escherichia coli, Shigella sonnei and Shigella flexneri, as well as Salmonella minnesota had a narrow range: 1.5-9 germs pro pg of mucin. The bacterial ligand of the binding is certainly not a polysaccharide as proved by the uniform binding of the R-mutant series of S. sonnei and S. minnesota. On the basis of inhibition tests by an outer membrane protein fraction, the ligand may be a common outer membrane protein of the enteric bacteria. The outer membrane proteins encoded by the Shigella-EIEC invasivity plasmids do not take part in this binding. The inhibition by killed bacteria or by their culture supernatants of mucin binding of heterologous species may suggest a non-species specific common ligand, too. Similarly to the mucin utilization, the binding ability also seems to be a general phenomenon among the enteric bacteria.

Animals↗

Isoelectric focusing of hog cholera virus.

The isoelectric point of Hog Cholera virus (Alfort strain) has been determined (pHi = 4.8). A clear stimulation of its infectivity can be recorded at this pH. The isoelectrophoretic profiles of strains isolated in the field show no significant differences.

Animals↗

Natural infection of pigs with bovine viral diarrhea virus and its differential diagnosis from hog cholera.

Natural infection of pigs with bovine viral diarrhea virus (BVDV) through contact with infected cattle has caused problems in diagnosing hog cholera (HC). Low cross-reacting serum antibody titers against HC caused by BVDV infection were found in clinically normal pigs as well as those suspected of having HC. Bovine viral diarrhea virus was isolated from specimen tissues and initially identified as HC virus (HCV), using the fluorescent antibody cell culture technique. Additional cell cultures, as well as pig and calf trials, were necessary to identify it as BVDV. The isolate caused clinical signs of illness in the calves, whereas the pigs remained healthy. Bovine viral diarrhea virus may be detected in tissue sections or isolated in cell cultures and confirmed as HCV, using the HC fluorescent antibody conjugate. Laboratories performing the neutralization test for HC should use discretion when interpreting HC titers unless BVD titers are determined on the same serums.

Animals↗

Kinetics of the inhibition of hog kidney D-amino acid oxidase by short-, medium- and long-chain fatty acids.

Various fatty acids were studied in vitro as inhibitors of pure hog kidney D-amino acid oxidase by means of a spectrophotometric peroxidase-coupling method using D-methionine as a substrate. All the fatty acids tested behaved as substrate-competitive inhibitors of the enzyme. The affinity of the saturated aliphatic acids for D-amino acid oxidase decreased from pentanoate (5:0; Ki = 220 microM) to laurate (12:0; Ki = 675 microM), then rose to a maximum with stearate (18:0; Ki = 36 microM), suggesting the presence of a site in the active center of the enzyme that accepts long-chain fatty acid alkyl groups. Unsaturation did not further increase the affinity of the fatty acid for this binding site.

Animals↗

Morphology of normal and osteochondrotic porcine articular-epiphyseal cartilage. A study in the domestic pig and minipig of wild hog ancestry.

Osteochondrosis is a generalized skeletal disorder that affects the growth cartilage in the growing domestic pig but not in the minipig of wild hog ancestry. In the present study, we compare the ultrastructure of the articular and epiphyseal growth cartilage in the domestic pig with that in the minipig. The domestic pigs had areas of enlarged epiphyseal cartilage with chondronecrosis, which had caused focal impairment of the endochondral ossification, with retention of cartilage in the subchondral bone. Areas of chondronecrosis close to blood vessels were found in the resting zone, with no evidence of thickened cartilage or impaired ossification. The chondronecrosis was surrounded by chondrocytes, organized in small clusters, containing many lipid droplets. The vascular channels adjacent to the chondronecrosis contained degenerated blood vessels. The minipigs showed no areas of enlarged epiphyseal cartilage. A few dead chondrocytes could be seen close to vascular channels which contained morphologically normal blood vessels. We conclude that restricted perivascular chondrolysis may occur in the pig without the presence of vascular degeneration and without progressing to osteochondrosis.

Animals↗

A blocking ELISA to differentiate hog cholera virus antibodies in pig sera from those due to other pestiviruses.

The blocking ELISA technique was extended to comparative serology by using 3 different pestivirus strains: Hog cholera virus (HCV) Alfort strain propagated in PK15 cell line, Border disease virus (BDV) Aveyron strain in PK15 and BVD NADL** strain in fetal calf kidney (FCK) primary cells. Rabbit antisera to the Alfort HCV strain and Aveyron BDV strain were raised for use in the test. A bovine hyperimmune serum to BVD virus was also used for detecting antibodies specific to BVD virus. The ELISA was compared with the neutralisation test on various groups of field and experimetnal porcine sera. The results obtained with the ELISA were well correlated with the neutralisation test. Therefore the ELISA may be recommended as a differential serological test between HCV and other pestivirus antibodies in pig sera.

Animals↗

Antigenic comparisons of hog cholera virus isolates from Europe, America and Asia using monoclonal antibodies.

Nineteen monoclonal antibodies (MAbs) with specificity for hog cholera virus (HCV) were prepared. They were used in an immune binding (peroxidase linked) assay to determine the reaction patterns of HCV isolates from Europe, Brazil, USA, Japan and Malaysia, as well as laboratory reference strains of the virus. A further panel of 17 MAbs raised against bovine virus diarrhoea virus (BVDV) was included in the study, together with 5 MAbs raised against a non-HCV pestivirus of porcine origin. All the MAbs were also tested against representative strains of BVDV and border disease virus. Six MAbs were HCV-specific, reacting with all isolates of HCV and none of the ruminant viruses. Among the other HCV MAbs geographical variation in reaction patterns was observed. There was evidence of antigenic distinction between recent European isolates, and archive material originally isolated more than 10 years ago.

Animals↗

The lapinized Chinese strain of hog cholera virus (HCV) previously adapted in minipig kidney (MPK) cell line cultures, would also protect pigs to experimental infection with virulent HCV [corrected].

Two-month old piglets previously inoculated with different dilutions of the Lapinized Chinese (LC) strain of Hog Cholera Virus (HCV), adapted in a minipig kidney (MPK) cell line, resisted challenge infection with virulent HCV. All the animals remained healthy and the challenge virus was never recovered from any of them. In contrast, the pigs which served as controls for the challenging virus underwent the clinically lethal form of the disease and HCV was consistently recovered from their tissues.

Animals↗

Isolation and antimicrobial susceptibility of Campylobacter coli and Campylobacter jejuni from slaughter hogs.

Cultural examination of cecal contents from 109 market weight hogs slaughtered in Prince Edward Island during May-July 1988 yielded 62 isolates of Campylobacter coli and seven Campylobacter jejuni. A commercial latex agglutination test helped to confirm the identification of Campylobacter. When tested against four drugs: erythromycin, tetracycline, kanamycin and ampicillin, 11 isolates showed multiple resistance. Resistance to erythromycin was seen in 19% and 28.6% of Campylobacter coli and Campylobacter jejuni respectively. All the isolates were susceptible to nitrofurans, gentamicin and chloramphenicol.

Ampicillin Resistance↗

Development and properties of a cell culture produced vaccine for hog cholera based on the Chinese strain.

The Chinese strain of hog cholera virus (HCV) was adapted to suspension cultures of the established swine kidney cell line SK6. The strain designated "Cedipest", is produced on the basis of a seedlot system. The masterseed virus was identified in vitro and in vivo, and was found free from extraneous pig pathogenic viruses by repeated animal inoculation followed by appropriate serological tests. A distinct and reproducible relationship was ascertained between infectivity in vitro and protection. Pigs inoculated with 400-600 TCID50 of the Cedipest strain proved fully protected against challenge with greater than 100 pig LD50 of a virulent strain of HCV at 7 days and at 6 month post vaccination.

Animals↗

Identification of a folate binder in hog kidney.

A macromolecular binder of folic acid (pteroylglutamic acid) and folic acid derivatives has been identified in extracts of hog kidney. With partially purified preparations, binding of [3H]pteroylglutamate was competed for by unlabeled pteroylglutamate, 5-methyltetrahydrofolic acid and its triglutamate derivative, by tetra- and dihydrofolic acid, and by N-10-formyltetrahydrofolic acid. The partially purified extract did not bine [3H]methotrexate nor could methotrexate or 5-formyltetrahydrofolic acid compete for [3H]folic acid-binding sites. The rate of binding of pterolyglutamate at 37 degrees was approximately 3%/s, was independent of pteroylglutamate concentration, and was essentially irreversible between pH 6.0 and 9.0. Below pH 6.0 binding was reversible, and at pH 3.5 the folic acid-binder complex completely disassociated. Based upon Sephadex gel filtration, the molecular weight of the folate-binder complex is 35,000 to 40,000. Binding activity was unaffected by pretreatment with ribonuclease or deoxyribonuclease but was completely destroyed by trypsin. The initial, unfractionated extract showed gamma-glutamyl carboxypeptidase (conjugase) activity which was lost in subsequent steps of purification of the folate binder.

Animals↗

The Cl- channel in hog gastric vesicles is part of the function of H,K-ATPase.

Hog gastric vesicles showed Cl- conductance when treated with Cu2+-o-phenanthroline, an S-S cross-linking reagent. An IgG monoclonal antibody caused dose-dependent inhibition of Cl- conductance that had been induced by S-S cross-linking. The antibody did not cause intervesicular aggregation, as determined by measurement of vesicle size. These results show that Cl- conductance, the stimulation and inhibition of which are regulated reversibly by S-S----2SH transformation, is due to native, physiological channels. The antibody also dose dependently inhibited the activities of H,K-ATPase and p-nitrophenyl phosphatase in gastric vesicles, but did not inhibit Na,K-ATPase obtained from dog kidney. Immunoblotting with the antibody of vesicle proteins solubilized in sodium dodecyl sulfate-polyacrylamide gel showed that the antibody binds to a 95-kDa subunit of H,K-ATPase and its dimeric 180-kDa polypeptide. The antibody-binding sites of H,K-ATPase activity and the Cl- channel for the inhibition were present on the external (cytosolic) surface of the transmembraneous ATPase. A gastric antisecretory compound, 2-methyl-8-(phenylmethoxy)imidazo[1,2 alpha] pyridine-3-acetonitrile (SCH 28080), competitively bound to the high affinity site of K+ on the internal (luminal) surface of H,K-ATPase, and its half-maximal inhibitory concentration for H,K-ATPase activity in tight vesicles was 0.2 microM in the presence of valinomycin. SCH 28080 also dose dependently inhibited opening of Cl- channels by S-S cross-linking, the regulatory site being present on the cytosolic side and more internally than the antibody binding site. The half-inhibitory concentration of SCH 28080 was 0.3 microM. The present results with the antibody and SCH 28080 indicate that the Cl- channel is part of the function of H,K-ATPase.

Adenosine Triphosphatases↗

[Molecular characterization of the hog cholera virus].

The molecular biology of hog cholera virus (HCV) was studied. After reverse transcription of the viral RNA the cDNA was cloned in the expression vector lambda gt11. HCV clones were identified using antibodies as a probe. Partial sequencing of one HCV-derived cDNA clone revealed a high degree of homology to a portion of the bovine viral diarrhea virus (BVDV). One goal of these studies is the preparation of defined, effective and safe vaccines against pestiviruses.

Amino Acid Sequence↗

Effects on platelets and on the clotting system of four glycosaminoglycans extracted from hog mucosa and one extracted from aortic intima of the calf.

A commercial heparin preparation, a heparin fraction with a molecular weight of 12,000 Daltons, heparan sulfate, dermatan sulfate obtained from hog mucosa, and mesoglycan, an heparinoid obtained from calf aortic intima were investigated. Commercial mucous heparin had a stimulatory effect on platelet aggregation induced by ADP, while the others failed to do so. Dermatan sulfate had a dose dependent inhibition and commercial mucosal heparin, a dose dependent stimulation, on serotonin release induced by ADP. Both the commercial mucosal heparin and dermatan sulfate showed an inhibition and the other glycosaminoglycans (GAGs) a negligible effect on collagen induced platelet aggregation. The collagen induced serotonin release was clearly reduced by all GAGs; heparan sulfate had this activity only at the highest doses used. Commercial mucosal heparin produced the highest activity on clotting systems as measured by activated partial thromboplastin time, while mesoglycan had the strongest anti-factor Xa specific activity as measured by a clotting assay. Dermatan sulfate was the weakest on both assays. When we injected intravenously an equivalent amount (about 60 mg) of heparin fraction, heparan sulfate, dermatan sulfate and mesoglycan in three different volunteers with an interval of 20 days after each injection, we had an immediate platelet factor 4 (PF4) release only with heparin fraction, heparan sulfate and mesoglycan. Heparin fraction and mesoglycan, in spite of having a wide discrepancy in anticoagulant effect, caused almost the same PF4 release. GAGs which can neutralize PF4 and which can also have specific anti-factor Xa activity could represent a great advantage in thrombosis prophylaxis.

Animals↗

A study on the susceptibility of minipig kidney (MPK) and rabbit kidney (RK13) cell line cultures to the lapinized Chinese strain of hog cholera virus.

The susceptibility of two established cell lines of pig (MPK = minipig kidney) and rabbit (RK13 = rabbit kidney) origin to the lapinized Chinese (LC) strain of hog cholera virus (HCV) was studied. Spleen cells from rabbits infected with the virus under study were inoculated to cell cultures of either MPK and RK13 cells and subsequent passages were made by culturing the trypsinized infected cells with the normal cells. Only the MPK cell line appeared to be susceptible to virus replication. Since no cytopathic effects (CPE) were observed, the presence of the viral antigen in the inoculated cultures was detected by immunofluorescence tests. The virulence of the virus for rabbits was enhanced after its cultivation in MPK cell cultures. When the MPK cell culture system adapted virus was tested in neutralization trials in the presence of an HCV reference immune serum it was found that the virus did not modify its antigenic structure in any extent. Finally, the culture adapted virus appeared to be more immunogenic for rabbits than the original rabbits adapted virus. Based on these results, it seems reasonable to suggest the use of MPK cell line for the propagation of the LC strain of HCV as an alternative to the use of rabbits for the preparation of HCV vaccine.

Animals↗

Hog cholera virus: art and facts.

The aim of this review is to summarize the informations accumulated during the two last decades on hog cholera virus, a member of the pestivirus genus. Different aspects concerning the virus-host cell interactions, and the structural and antigenic properties of the virion itself are successively analyzed. Despite numerous works, many basic informations are still lacking, which can be explained by the difficulties inherent to the study of this virus.

Classical Swine Fever Virus↗

Pathogenesis and epidemiology of hog cholera.

A review is given on the infection with hog cholera (HC) virus resulting in various courses due to variabilities of the virus and the porcine host organism. Mechanisms by which the virus causes prenatal death or postnatal disease in pigs are considered. Epidemiological features relating to various courses of infection and disease patterns resulting from prenatal or postnatal contacts with the virus are discussed.

Animals↗