Infectivity for mice of avian influenza A viruses of H7N7, H5N3 and H2N2 subtypes isolated from migratory waterfowls in San-in District, Western Japan.
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Biomedical subjects
Publications and source records attributed to K Otsuki.
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Ten strains of avian infectious bronchitis virus (IBV) were studied serologically by cross-neutralization test using rabbit and chicken immune sera. With the chicken sera all 10 IBV strains were antigenically related. In particular, anti-KH serum neutralized all heterologous strains except of the Ishida strain; Nerima strain was neutralized by all antisera except of anti-Ishida serum. Most cross-reactions were less or more heterologous, thus all 10 IBV strains seemed to belong to one serological type. Using rabbit sera, all strains except of Connecticut A-5968, cross-reacted with certain other strains. Most cross-reactions were partially heterologous showing one-way-relationship; heterologous relations were observed less frequently than with chicken sera.
From November 1982 to March 1983, winter migratory waterfowls of some species staying in San-in District, Western Japan, were surveyed for influenza virus at five stations. A total of eight influenza A viruses were isolated from 354 faeces samples of whistling swans; in contrast, no virus was isolated from any sample of 261 black-tailed gulls, of 113 pintails and of 10 mallards. Five of eight isolates belonged to human pandemic subtype H2N2, two isolates belonged to fowl plague subtype H7N7, and the remaining one to subtype H4N6.
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Certain species of winter migratory waterfowl in San-in District, western Japan, were surveyed for influenza virus from November 1983 to March 1984. Faeces of the waterfowl were collected regularly at five stations. Eleven influenza A viruses including H5N3 and H10N4 subtypes were isolated from 450 faecal samples from whistling swans, 28 viruses including H2N2 and H10N4 subtypes were isolated from 362 faecal samples from pintails; and subtype H13N6 was isolated from 240 faecal samples of black-tailed gulls.
One of 16 raw milk isolates of Yersinia enterocolitica and Y. intermedia produced heat-stable enterotoxin (ST) in milk at 25 degrees C but not at 4 degrees C after 21 days of incubation. A catabolite repression of ST synthesis by the lactose-fermenting strain of Y. enterocolitica was observed when 4.6% lactose was added to trypticase soy broth. However, the lactose-fermenting strain was killed by acid produced by lactose fermentation in milk and did not produce ST in milk with the pH adjusted to neutrality. This study suggested that lactose and fat in milk are not the fundamental inhibitors of ST synthesis by Y. enterocolitica and that repression of ST synthesis may be related to other components.
Lipopolysaccharide (LPS) of a reference strain of Yersinia enterocolitica serovar O15, Ye614, was examined for its chemical structure and antigenicity. The O15 LPS was partially hydrolyzed with diluted acetic acid and fractionated on a Sephadex G-50 column. Only one peak was detected at fractions where the R-core oligosaccharide of a Y. enterocolitica rough strain was eluted. The chemical compositions of this oligosaccharide and the O15 LPS were similar to those of the R-core oligosaccharide and the rough LPS, respectively. The O15 LPS was antigenically identical to the rough LPS in immunodiffusion tests. These results indicate that the O15 LPS of Y. enterocolitica is rough LPS and that antigenic factor O15 is a rough antigen.
In Shimane Prefecture, Japan, 175 Yersinia enterocolitica isolates from 6,479 stools and 347 appendixes of patients were separated into biotype 3B (VP-, sorbose-, inositol-) serotype O3 phage type II (22 isolates); biotype 4 (84 isolates), biotype 4 (ornithine-) (16 isolates) and biotype 4 (maltose-) (9 isolates) of serotype O3 phage type VIII, biotype 2 serotype O9 (1 isolate) and biotype 1 (43 isolates). This may be the first documentation of isolation of Y. enterocolitica biotype 3B serotype O3 phage type II from patients. Twenty-three Yersinia pseudotuberculosis isolates from stools of 23 patients were grouped into serotypes IB (13 isolates), III (2 isolates) and IVB (8 isolates). Clinical manifestations were more numerous in Y. pseudotuberculosis infections than Y. enterocolitica O3 infections. Y. enterocolitica O3 was more frequently isolated between summer and autumn but Y. pseudotuberculosis and Y. enterocolitica biotype 1 were isolated between winter and spring. Secondary infection with Yersinia among close family members was frequent. Y. enterocolitica O3 and Y. pseudotuberculosis infected persons excreted 10(4) to 10(9) viable cells per g stool during 27 and 19 days, but Y. enterocolitica biotype 1 strains were detected only when using enrichment techniques. Serological responses were not observed in under 2 year old persons infected with Y. enterocolitica O3.
Fattening pigs were inoculated with the human pathogenic strains of Yersinia enterocolitica, biovar 4 serovar 3 phagovar 8 and biovar 2 serovar 5.27. Each pig received 2.6 X 10(9) organisms by catheter into the stomach and thereafter 10% sodium bicarbonate solution, by the same route. The serovar 3 strain became established in the intestines of four out of six 11-week-old pigs and in all three 24-week-old pigs. Serovar 5.27 established in the intestines of all four 10-week-old pigs. In these pigs, both serovars were excreted in the feces at 10(2)-10(6) cells g-1 for a few weeks. However, the serovar 5.27 was excreted in the feces at less than 10(3.3) cells g-1 for 2 weeks by 24-week-old pigs. Horizontal transmission with the serovar 5.27 strain was observed on the 5th-11th day, and the bacteria were present in the intestines during the 2nd week. However, the serum titers to O-agglutinin were 1/10 and less, but in one pig, the titer was 1/40 against serovar 5.27 strain.
Yersinia enterocolitica biovar 4 serovar 3 and biovar 2 serovar 5.27 failed to establish in the intestines of pigs when challenged with the homologous or heterologous strains. After inoculation, the serum O-agglutinin titers were 1/10 or less and were not boosted by challenge with either serovar. Y. enterocolitica were not recovered in any specimens at the time of slaughter.
Twelve strains of Yersinia enterocolitica and 4 strains of a Y. enterocolitica-like organism (Y. intermedia) were isolated from 374 pooled raw milk samples. Growth occurred in a cold enrichment system using phosphate buffer solution (7 strains), peptone solution (5 strains) and yeast extract-casein-cystine broth (5 strains). Serovar 13,7 was most frequently isolated (8 strains) followed by serovars 5,27 (2 strains), 6,31, 7,8, 14 and 22 (each one strain). Two strains were untypable, serologically. Seasonal variation was of little consequence, but the regional incidence was an important feature of the bacteriological findings. Autoagglutination tests gave negative results with all isolates. Although so-called "clinical strains" were not identified, the relationship of the strains of Yersinia to milk-borne infections in humans cannot be excluded.
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A total of 93 Yersinia spp. were isolated from 68 of 252 dogs in Shimane Prefecture, Japan. The Yersinia spp. were 70 Yersinia enterocolitica isolates from 50 dogs, 2 Yersinia frederiksenii isolates from 2 dogs, 5 Yersinia intermedia isolates from 5 dogs, and 16 Yersinia pseudotuberculosis isolates from 16 dogs. Fifteen of 70 Y. enterocolitica isolates belonged to biotype 4 serotype O3 phagotype 8 (6 isolates), biotype 4 (maltose negative) serotype O3 phagotype 8 (4 isolates), biotype 3 (Voges-Proskauer and sorbose negative) serotype O3 phagotype 9 (2 isolates), and biotype 2 serotype O5.27 (3 isolates). The other 55 Y. enterocolitica isolates belonged to biotype 1 and were classified into serotypes O4, O5, O7 .8, O9, O10 , O12 , O13 .7, O14 , O15 , and untypable. Sixteen Y. pseudotuberculosis isolates belonged to serotypes IB (two), IIB (two), IVA (two), IVB (three), VA (five), and untypable (two). These isolates were mostly detected in cold months. Y. enterocolitica serotype O3 and Y. pseudotuberculosis were isolated more frequently from puppies. Y. enterocolitica serotype O3 was recovered at less than 10(7.0) cells per g of the jejunal-to-rectal contents and at less than 10(2) cells per g of mesenteric lymph nodes. Y. pseudotuberculosis was recovered at less than 10(4.3) cells per g of the cecal-to-rectal contents and at less than 10(2.9) cells per g of mesenteric lymph nodes. Almost all strains of Y. enterocolitica biotype 1, Y. intermedia, and Y. frederiksenii were recovered at less than 10(2) cells per g of gastric-to-rectal contents.
Yersinia pseudotuberculosis serogroup III is most frequently detected in healthy pigs. Y. pseudotuberculosis strains were divided into two biogroups on the basis of their fermentation of melibiose. Melibiose-fermenting strains were distributed in all the hosts and serogroups examined. Melibiose-nonfermenting strains were limited to serogroup III isolated from healthy pigs. Autoagglutination and calcium dependency were not related to fermentation of melibiose. Melibiose-fermenting strains seem to be virulent for mice, but melibiose-nonfermenting strains showed no virulence for mice.
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Yersinia spp. (741 strains) were recovered from 81% of 48 surface water samples collected over a 12-month period from four rivers in Matsue, Japan. The precipitation methods with FeCl3 or Kaolin and the cold enrichment method with Peptone-Mannitol-Phosphate buffer solution were used for recovery. Isolates belonged to Yersinia enterocolitica (Ye) (133 strains), Yersinia intermedia (511 strains), Yersinia frederiksenii (57 strains), Yersinia kristensenii (10 strains) and X2-like organisms (30 strains). Thirty colonies of Ye O3 biotype 3 per ml surface water may relate to the drainage containing 2 X 10(4) Ye O3 biotype 3 per ml, from the piggery that raised Ye O3 biotype 3-positive pigs. There was the negative interrelation between the incidence of isolation of Yersinia spp. and the environmental- and water temperatures. This may be the first documentation of isolation of Ye O3 from surface water.
Intestinal contents of tufted ducks and faecal samples from two other species of migratory birds were examined for Yersinia species of organisms. From 296 tufted ducks Yersinia species were isolated from 10 (3.4 per cent), from 223 swans Yersinia species were recovered from eight (3.6 per cent) but none were recovered from the faeces of 121 pintail ducks. Since the isolates differed from each other in their biochemical characteristics and, or, O serotypes, yersinia infection in these migratory waterfowl seemed to be sporadic.