Effect of selective vaccination on a decrease in the rate of hepatitis B virus-positive Japanese first-time blood donors.

The government of Japan started a selective vaccination programme to prevent mother-to-infant infection by hepatitis B virus (HBV) since January 1986. The effect of the programme on first-time blood donors has not been examined in detail. Data of first-time blood donors aged 16-25 years from 1996 to 2007 were extracted from the Japanese Red Cross (JRC) donors' database. Principal component analysis (PCA) was used to visualize the birth-year-dependent group of rate of HBV-positive donors. According to the birth of year, donors were divided into four groups by PCA. After the start of the programme, donors born in 1986-1989 comprised a single group. Before the start of the programme, three groups (1980, 1981-1984 and 1985) were identified. Although a significant time-dependent decrease in the rate of HBV-positive donors was observed before the start of the programme, a significant difference in the rate of HBV-positive donors was observed around the start of the programme by regression analysis for 16-19-year-old first-time blood donors. The selective vaccination programme has been effective to prevent the vertical transmission of HBV from the analysis of first-time blood donors. On the other hand, vaccination of blood donors should be considered to reduce the risk of post-transfusion HBV infection, because the horizontal transmission increases in HBV-positive blood donors.

Superiority of minipool nucleic acid amplification technology for hepatitis B virus over chemiluminescence immunoassay for hepatitis B surface antigen screening.

BACKGROUND AND OBJECTIVES: The Japanese Red Cross (JRC) have developed a fully automated multiplex (MPX) nucleic acid amplification technology (NAT) system for hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus 1 (HIV-1). This is used to test serologically negative blood units from volunteer, non-remunerated donors. The system utilizes a 50-sample pool for NAT screening with an input volume of each pool. This results in a significantly higher sensitivity for hepatitis B than that seen with highly sensitive hepatitis B surface antigen (HBsAg) testing. MATERIALS AND METHODS: From 1 February 2000 to 15 October 2001, over 11 million donations, which were serologically negative, were tested using the MPX NAT system. Donations found to be HBV DNA positive were further tested by using the chemiluminescence immunoassay (CLIA). RESULTS: Out of 181 HBV DNA-positive donations, 96 (53%) and 76 (42%) were negative by individual enzyme immunoassay (EIA) and CLIA testing, respectively. CONCLUSIONS: The sensitivity of the 50-sample pool MPX NAT system was higher than that of individual HBsAg screening by CLIA. By adopting this NAT-screening system, the JRC has improved the safety of the blood supply and maintained supply across Japan.

Analysis of distribution of Campylobacter jejuni and Campylobacter coli in broilers by using restriction fragment length polymorphism of flagellin gene.

The incidence of Campylobacter jejuni and Campylobacter coli in broiler farms was 33.9% (19/56). C. jejuni-positive flocks accounted for 20.0% (17/85) and C. coli-positive ones was 4.7% (4/85). There were 14 patterns (fla type) of restriction fragment length polymorphism (RFLP) of flagellin A gene among these 22 strains of C. jejuni and C. coli including the standard strain C. jejuni ATCC 33560. Different fla types of Campylobacter were isolated from broilers in different growing cycles on the same farms. Four strains of C. jejuni were isolated from four breeder farms and four fla types of C. jejuni were detected from their progenies reared on growing farms. Three fla types of C. jejuni detected from the progenies were different from those of each breeder. Also, the other three fla types of C. jejuni were detected from different progenies of each growing farm during the next growing cycle. These findings indicate that the RFLP analysis may contribute to epidemiological studies of C. jejuni and C. coli contamination of broilers and suggest the risk of contamination with different types of Campylobacter in every growing cycle of broilers on the farm even on the same farm. They also supported that there was little likeliness of the vertical transmission of C. jejuni and C. coli from breeders to broilers.

Direct detection of Campylobacter jejuni in chicken cecal contents by PCR.

Campylobacter jejuni in chicken feces was detected by PCR and Southern blot hybridization (SBH). The detection limits of C. jejuni in chicken feces were 34,000 cells by PCR and 340 cells by SBH. Some cecal contents of chickens up to 3 weeks old were C. jejuni positive by SBH whereas all of them were negative by PCR. Two of 51 cecal contents of 18-day-old chicken embryos were C. jejuni positive by PCR and SBH; but, C. jejuni were not isolated from the samples by conventional culture with selective enrichment.

A survey of Campylobacter jejuni in broilers from assignment to slaughter using DNA-DNA hybridization.

A survey of Campylobacter jejuni in the cecal contents of broilers raised on a farm was carried out by the DNA-DNA hybridization method from the day of assignment to slaughter at about 1-week intervals. C. jejuni was detected in chickens as early as 1 week of age, and was widely detected at each week of age throughout the growing period. In addition, of 20 chickens tested just after assignment, 7 (35%) were C. jejuni positive. It is suggested that newly introduced chickens may have already been contaminated with C. jejuni. The hybridization method was able to detect C. jejuni in the chickens from the day of assignment to 3 weeks of age, but C. jejuni was never detected in the same chickens by the enriched culture method. Therefore, it is suggested that the hybridization method is more sensitive than the enriched culture method.

Application of a DNA-DNA hybridization method for detection of Campylobacter jejuni in chicken feces.

A direct colony hybridization method was used for the detection of Campylobacter jejuni in chicken feces. The biotin-labeled DNA prepared from the whole genome DNA of C. jejuni subsp. jejuni ATCC 33560 reacted well with homologous DNA and slightly with C. coli DNA. The method with the probe was found to be sensitive enough to detect a small number (10(2) CFU/g) of C. jejuni in chicken feces which contained a large number of background flora. It was suggested that this simple and sensitive method was useful for a wide survey of C. jejuni.

Production and characterization of serovar-specific monoclonal antibodies to serovars 4, 8, and 9 of Mycobacterium intracellulare.

Serovar-specific monoclonal antibodies against Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex serovars 4, 8, and 9 were prepared. Nine, four, and one monoclonal antibodies, respectively, to the serovars were prepared by the usual cell fusion technique. All nine monoclonal antibodies to serovar 4 were monospecific for their homologous serovar and reacted with several native glycopeptidolipids (GPLs) and one major deacylated GPL from the homologous serovar. One of the four monoclonal antibodies to serovar 8 seemed to be monospecific for its homologous serovar, but the other cross-reacted with serovar 6 because serovar 6 organisms contain the same components as does the major deacylated GPL from serovar 8. One monoclonal antibody to serovar 9 was monospecific for its homologous serovar and reacted with one of the two major deacylated GPLs from this serovar. These antibody preparations proved useful for serovar identification.

Light and electron microscopic observations on granulomatous lesions in pigs dosed with Mycobacterium intracellulare.

The histology and ultrastructure of the granulomatous lesions were studied in 18 pigs dosed orally with Mycobacterium intracellulare serotype 8. The pigs were killed 2, 4, 6, 8 and 12 weeks after dosing. Histologically, initial granulomatous lesions were seen in the tonsils 6 and 8 weeks after dosing. Initial or more advanced granulomatous lesions were observed in the tonsils and lymph nodes (mandibular, jejunal, ileocolic and superficial inguinal) 12 weeks after dosing. The initial granulomatous lesions appeared as slight proliferations of epithelioid cells and multinucleated giant cells. Advanced granulomatous lesions were nodular proliferations of epithelioid cells and multinucleated giant cells (epithelioid-cell nodule). In more advanced granulomatous lesions, the epithelioid-cell nodules increased in size and showed caseation necrosis, calcification and fibrosis. Ultrastructural examination revealed that the epithelioid-cell nodules consisted of mononuclear cells, epithelioid cells and multinucleated giant cells. In the caseous foci of the epithelioid-cell nodules, an electron-dense amorphous material with collagen fibres was deposited in the intercellular spaces. Although a small number of acid-fast organisms were observed histologically, they could not be detected ultrastructurally. The histological and ultrastructural findings of the present cases are compared with those of pigs infected with M. avium and with human lesions of tuberculosis and sarcoidosis.

Relationship between serotype and certain biological and biochemical characteristics of strains of the Mycobacterium avium and Mycobacterium intracellulare complex.

Some relationships among Schaefer's serotypes and biological and biochemical characteristics were observed in strains of the Mycobacterium avium-Mycobacterium intracellulare complex. Strains belonging to serotypes 2 and 16 lacked the capacity to utilize n- and iso-butanols as the sole source of carbon in the presence of ammoniacal nitrogen. However, strains of serotype 2 grew at 45 C and lacked arylsulfatase activity (after 14 days), whereas strains of serotype 16 failed to grow at 45 C and showed positive arylsulfatase activity. Strains belonging to serotypes 8, 9, and 15 grew at 45 C, utilized n- and iso-butanols, and showed arylsulfatase activity.