BACKGROUND: It is believed that bovine neutrophils contain several peptides and protein which exhibit antimicrobial activity against microorganisms such as fungi and bacteria. Objectives: The purpose of this study was to isolate and purify a potential antibacterial protein from bovine neutrophil and test its anti bacterial activity. Methods: Neutrophils were isolated from bovine blood using dextran sedimentation and centrifugation on Ficoll-Hypaqe. Cell viability is examined by trypan blue dye exclusion method. Protein extract was then dialyzed and applied onto ion exchange chromatography for further purification, and its potential cytostatic activity was examined against Staphylococcus aureus, Aeromonas hydrophila and Yersinia ruckeri. Results: Viability of isolated neutrophil was over 95%, chromatographic results and SDS-PAGE analysis exhibited that neutrophil cytosolic proteins were fractionated so that the purification of P7/P23 was almost 60% in the first step and purification was completed in the second phase. Using calibration curve according to molecular mass markers, the relative molecular mass of P7 and P23 determined as 7 kDa and 23 kDa, respectively. Also, results showed that this protein has antibacterial activity and has higher bactericidal activity against Y. ruckeri. Conclusions: It could be concluded that purification of P7/P23 sustains its biological activity and has wide range antibacterial activity. Moreover, taking all data into account may suggest that the cytostatic activity of neutrophil, to some extent, results from P7/P23 protein which is abundant in the cytosole.

A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose. For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions. The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride. The neutrophils (2 X 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ([32P]SA; 2 X 10(8)) inskimmed milk samples (2.5% final concentration) prepared from 4 cows. This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals. The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1%. Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation. The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern. The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis. The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.

The first outbreak of aujeszky's disease (AD) was identified from piggery located at the southern part of Korea in July, 1987. This piggery suffered from a significant economic loss caused by unexpected piglet mortality and reproductive failure. Etiologic viral agents were isolated from tonsil and spleen of the infected piglets, and the isolates produced a typical cytopathic effect of herpesvirus with giant cell formation when inoculated in many different cells. Subsequently the field isolates were characterized as suid herpesvirus I by cross-neutralization test and indirect fluorescence assay utilizing specific monoclonal antibody, and proved to be a pathogenic strain of AD virus(ADV).

Mating experiments were performed to elucidate the distribution of perfect state of Microsporum gypseum complex originating from animals and soils in Korea. A total of 30 strains of M. gypseum complex, composed of 16 from animals and 14 from soils, mated with the tester strains of Nannizzia incurvata, N. gypsea and N. fulva. Among 30 strains of M. gypseum complex examined, 16 strains (53.3%) were N. incurvata, 13 strains (43.3%) N. gypsea and 1 strain (3.3%) N. fulva. Among 13 strains of dog isolates, 8 strains (61.5%) were N. incurvata and 5 strains (38.5%) N. gypsea. And each strain isolated from Korean native goat, monkey and albino rat was N. gypsea. Among 14 strains of soil isolates, 8 strains (57.2%) were N. incurvata, 5 strains (35.7%) N. gypsea and 1 strain (7.2%) N. fulva. And - mating type was more frequently obseved than + mating type in the strains of N. incurvata and N. gypsea. By this study, N. fulva was isolated the first time in Korea.

This paper deals with the distribution of Salmonella (S) infection on 4 herds in Kyungju and Taegu during the period from May to October 1986. Isolated Salmonella were examined for serotypes, antimicrobial drug resistance and detection of R plasmid. From 4 herds, 67 Salmonella were isolated from 51 samples (1.1%), and their serovar strains were S. typhimurium 6, S. derby 5, S. infantis 4, S. bareilly 4, S. dublin 3, S. anatum 2, S. montevideo 2 and untypable 41. In 4 herds, the incidence of drug resistance was 57.7-100% and transfer frequency of conjugative R plasmid was 96.1-100%.

This paper dealt wih the distribution of Shigella spp. on 5 piggeries in Taegu and Kyongbuk during the period from August to October 1987. Isolated Shigella were examined for serogrouping, antimicrobial drug resistance and detection of R plasmid. Genetic properties of R plasmid in Shigella were examined for fertility inhibition (F1) and gel electrophoresis was performed for the isolation of plasmid DNA. Of total 2,978 samples from 5 piggeries, 82 strains (2.8 %) of Shigella spp. were isolated from 82 samples. The isolated strains were identified as S. dysenteriae (60 strains), S. flexneri (20 strains) and S. sonnei (2 strains). Of the 82 strains examined 67 (95.1 %) were resistant to one or more antibiotics, such as ampicillin (Am), chloramphenicol (Cm), kanamycin (Km), nalidixic acid (Na), rifampicin (Rf), streptomycin (Sm), sulfademethoxine (Su), and tetracycline (Tc) and higher resistance to Su (90.2 %), Sm (63.4 %) and Tc (63.4 %). Of the 78 resistant Shigella strains 26 (33.3 %) harbored conjugative R plasmids and the transfer frequency of Sm (50.0 %), Cm (33.3 %) resistance was much higher than that of the other drug resistance. The most common resistant patterns were SmSuTc, Su and AmSmSuTc. Out of the 26 Shigella R plasmids examined for Fi, 14 (53.8 % were Fi + and the remainder were Fi-. The plasmid DNA profiles in Shigella spp. (9 strains) isolated from pigs were confirmed as being 2 to 9 fragments by the gel electrophoresis. Their molecular size ranged 2.17 to 87.62 kilobase (Kb). All strains of Shigella spp. consisted in 15.4 Kb plasmids.

The present study was conducted to investigate the Bordetella bronchiseptica infection in Youngnam swine herds during the period of August 1986 to July 1987 and some properties of the organisms isolated from these Korean swine. B. bronchiseptica was recovered from 25 of 70 (35.7%) growing pigs of 4 to 10 weeks of age and from 12 of 13 (92.3%) herds. From 115 slaughter pigs, 58(50.4%) pigs were culture positive and the pigs from 13 of 14 (92.9%) herds were found to be infected with B. bronchiseptica. The majority of biochemical and cultural properties of B. bronchiseptica isolated from Korean swine were identical to those of the standard strain employed and some 97.6% of the isolates showed the characters of phase I organism on primary isolation.