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Biomedical subjects

N Kagiyama

Publications and source records attributed to N Kagiyama.

At least 37 records · Page 2Linked to original sources

Differences in susceptibility to peroral inoculation with Bacillus piliformis spores in rats and mice.

The median liver lesion producing doses of peroral inoculation with the spores of Tyzzer's organism RJ strain were 10(4. 3) in rats and 10(2. 7) in rats receiving prednisolone treatment for the provocation of Tyzzer's disease. In contrast to rats, liver lesions were detected in few mice inoculated perorally with 10(7) spores. In mice inoculated perorally with 10(7) spores, excretion of infective spores in the feces was detected only on day 1 postinoculation. On the other hand, no difference in susceptibility between rats and mice was detected upon intravenous inoculation with vegetative cells of the RJ strain. These results suggest that germination of the spores in the intestinal tract causes the difference in the susceptibility in rats and mice.

Animals↗

Determination of an antigen suitable for enzyme-linked immunosorbent assay of the antibody to Bordetella bronchiseptica in guinea pigs.

To establish an enzyme-linked immunosorbent assay (ELISA) technique for the serological diagnosis of infections caused by Bordetella bronchiseptica (B. bronchiseptica) in guinea pigs, the authors recently assessed the usefulness of three antigen preparations derived from the bacterial cell components: sonication antigen (S-Ag), cell surface antigen (C-Ag) and lipopolysaccharide antigen (L-Ag). The use of S-Ag for ELISA resulted in the most sensitive detection of the antibody to B. bronchiseptica from guinea pig sera immunized with killed bacteria and sera derived from naturally infected guinea pigs. Like C-Ag, S-Ag was highly specific, showing no cross-reactivity with Pasteurella multocida. Assessment of antibody formations in animals with experimentally induced infection using the three antigen preparations revealed that the antibody to S-Ag was formed earlier than antibodies to the other two antigen preparations following growth of the bacterium in the lungs. These results indicate that ELISA with S-Ag as an antigen is a useful tool for the serological diagnosis of infection by B. bronchiseptica.

Animals↗

[The reactivity of antigens of Mycoplasma pulmonis derived from mice and rats to naturally infected rat sera].

The reactivity of antigens of 4 mouse and 3 rat derived Mycoplasma pulmonis strains to 20 naturally infected rat sera was studied. The optical density values of the same serum by enzyme-linked immunosorbent assay using the 7 strains as the antigen revealed no marked difference among the strains. M. pulmonis antigens recognized by the antibodies were analyzed by the Western immunoblot method. The antigens with molecular weights of 92 K, 66 K, and 58 K were recognized in the 7 strains at a high frequency.

Animals↗

Genomic DNA analysis of rodent mycoplasmas.

Genomic DNA was compared between three typical species of rodent mycoplasmas, Mycoplasma pulmonis, M. arthritidis and M. neurolyticum, and between strains of these species. Each of the three species showed a distinct restriction endonuclease cleavage pattern of genomic DNA. The genetic heterogeneity of these species was revealed by total DNA hybridization as well. In addition, the restriction endonuclease cleavage pattern of genomic DNA was almost identical in three strains of M. pulmonis and in two strains of M. neurolyticum. The genetic hemogeneity among strains of the same species was revealed by total DNA hybridization as well. These data suggest that the genomic DNA sequence of each rodent mycoplasma species has a high degree of species specificity.

Animals↗

Expression of Mycoplasma pulmonis antigens in Escherichia coli.

The expression of Mycoplasma pulmonis antigen in Escherichia coli was investigated by cloning genomic DNA derived from M. pulmonis m 53, and the DNA fragment participating in antigen expression was identified. When the DNA library of M. pulmonis was screened by colony immunoassay using anti-M. pulmonis serum, 10 recombinant clones expressing seroreactive antigens were obtained. The recombinant plasmids isolated from these clones included 3.7-6.5 kilobase pair (kbp) DNA inserts, while all clones contained a common 2.3-kbp DNA fragment. Subcloning of initial DNA inserts showed that the common 2.3-kbp fragment is essential for antigen expression. Moreover, antiserum against the recombinant antigen generated from the 2.3-kbp DNA fragment recognized a native M. pulmonis antigen. The reactivity of this antiserum was absorbed specifically with M. pulmonis. These results suggest that the cloned 2.3-kbp DNA fragment codes an antigen specific to M. pulmonis.

Antigens, Bacterial↗

A DNA probe for specific detection of Mycoplasma pulmonis.

Mycoplasma pulmonis was specifically detected by using a 2.3 kilobase pair (kbp) cloned DNA fragment derived from M. pulmonis m 53 as a probe. This probe recognized 2.3-kbp DNA fragments of three M. pulmonis strains in Southern hybridization, while it did not hybridize with the DNA of M. arthritidis or M. neurolyticum. Determination of the sensitivity of the probe by dot hybridization revealed that 10 ng of M. pulmonis DNA was detected by a biotinylated probe and 1 ng of M. pulmonis DNA was detected by a radioactive probe.

DNA Probes↗

Production of Tyzzer's disease in rats by ingestion of bacterial spores.

Tyzzer's disease was produced in rats by peroral inoculation with spores of Bacillus piliformis (Tyzzer's organism) of rat origin. After ingestion of 10(6) spores necrotized lesions with intracellular bacterial propagation were seen in the intestines, liver and heart on days 2 to 14 postinoculation (p.i.). A number of B. piliformis were present within enterocytes of the cecum and colon. Infected cells were also seen in the liver, myocardium and intestinal muscle layers on days 3 to 7 p.i. Infective spores were found to be shed in feces during 3 to 10 days p.i.

Animals↗

Cross reactivity of Mycoplasma pulmonis and Mycoplasma arthritidis antigen strains to anti-Mycoplasma pulmonis antibody in the sera of Mycoplasma pulmonis infected rats.

Reactivity of Mycoplasma pulmonis (Mp) antigen strains to anti-Mp antibody in the sera of Mp infected rats was examined by enzyme linked immunosorbent assay (ELISA). Antibody titers to 7 kinds of Mp antigens were measured in the sera of 20 Mp isolated rats and 20 Mp free rats by ELISA and complement fixation test (CF test). ELISA showed that there was no difference in the antibody titer of the same serum among 7 Mp antigen strains employed, and the main cross reaction to anti-Mp antibody took place on the common recognition site (common antigen) in all the Mp antigens. The CF test suggested that the antibody titers largely differed due to the kind of Mp antigen strains, and the detection rate was between 0 and 60%, presumably due to the difference in the reactivity or binding ability of complements but not due to the difference in the cross reactivity of Mp antigen to anti-Mp antibody. When the cross reactivity of anti-Mp antibody to Mycoplasma arthritidis (Ma) antigen was examined in the sera of 33 Mp isolated rats, the CF test exhibited the negative results, but ELISA showed 4 positive cases to support the cross reactivity of anti-Mp antibody to Ma antigen.

Animals↗

Enzyme-linked immunosorbent assay for detection of serum antibody to CAR bacillus.

An enzyme-linked immunosorbent assay (ELISA) for detection of CAR bacillus antibody in rat sera was developed by Ganaway et al., in 1985 although the ELISA method was not described in detail. We investigated antigen preparation and test procedures of the ELISA using two strains of CAR bacillus which we isolated from a mouse (CB-M) and a rat (CB-R). Allantoic fluids containing 2.4 X 10(8)/ml of CB-M and 2.0 X 10(8)/ml of CB-R were washed with sterile phosphate buffered saline (PBS), resuspended in a 1/5 volume of sterile carbonate buffer (pH 9.8) and sonicated. Then 1/40 and 1/80 dilutions of CB-M and CB-R lysates in PBS, respectively, were used for antigen solutions of ELISA. Briefly, antibodies in sera are reacted with antigens coated on the surface of microtiter plates. The amount of horse radish peroxidase labeled protein-A or anti-rat IgG bound to the antigen-antibody complexes is measured on the spectro photometer at wave length of 492 nm. A total of 180 mouse and 205 rat sera were tested against both antigens. The optical density (OD) values of 140 mouse and 161 rat sera obtained from SPF mice and rats free from CAR bacillus infection were on the average 0.005 and 0.019, respectively. On the other hand, OD values of the sera collected from CB-M or CB-R infected animals ranged from 0.20 to 1.52. According to these results, the cut-off OD value for positive reaction was set at 0.1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Detection of Sendai virus antibody in mouse and guinea pig sera by an enzyme-linked immunosorbent assay with protein A.

An enzyme-linked immunosorbent assay using horseradish peroxidase (HRPO)-labeled protein A (P-ELISA) was established for detection of Sendai virus (SV) antibody in mouse and guinea pig sera. Sensitivity and specificity of P-ELISA were compared with those of ordinary ELISA using HRPO-labeled immunoglobulin G (IgG-ELISA) and the hemagglutination inhibition (HI) test. P-ELISA was 100 to 1,000 times more sensitive than the HI test for detection of the antibody in SV-naturally infected mice. P-ELISA and IgG-ELISA showed similar sensitivities for detection of the antibody in naturally infected mouse and guinea pig sera. A high specificity was demonstrated in P-ELISA with a cut-off optical density value of 0.2 (492 nm), while a non-specific reaction was observed when IgG-ELISA was used to both mouse and guinea pig sera at a low dilution (1:10-20). The antibody in rat sera was not detected by P-ELISA although it was realized by IgG-ELISA.

Animals↗

Pathogenicities of two CAR bacillus strains in mice and rats.

Two strains of CAR bacillus from a mouse (CB-M) and a rat (CB-R) which were passaged 11th in embryonated chicken eggs via the allantoic route were inoculated intranasally in ICR mice and Wistar rats. The histopathological changes and the localization of the CAR bacillus in the tracheas and lungs of these animals were investigated microscopically 2, 4 and 8 weeks postinoculation (PI). Histopathological changes similar to those in natural cases of CAR bacillus infection, showing severe peribronchial lymphoid cuffing, were first recognized 4 weeks PI. CAR bacillus was also found on the cilia of the respiratory epithelium. These histopathological changes were more remarkable in mice inoculated with CB-M and in rats inoculated with CB-R.

Animals↗

Microbiological monitoring in inbred mouse foundation stocks in Japan.

Microbiological monitoring on 128 inbred mouse foundation stocks consisted of common 10 inbred strains and inbred strains originated from outbred dd mice was performed by cooperation of 24 organizations. A total of 881 mice were divided into 647 conventional animals from 95 colonies and 234 barrier-sustained animals from 33 colonies. Three viral, one mycoplasmal, 6 bacterial, one fungal and 3 parasitic agents selected as monitoring microbes according to the proposed selection standards. Among conventional colonies, 84.2% were positive for at least one agent. The highest detection rate was 44.2% for S. obvelata, followed by P. pneumotropica and S. muris, P. aeruginosa, G. muris, Sendai virus, M. pulmonis, MHV and E. coli O115a, c: K (B). Of these agents, only one microbe, P. aeruginosa, was detected in barrier-sustained colonies (36.4%), thus the efficacy of barrier system for the microbiological quality control of the inbred mouse foundation stocks was actually demonstrated. The positive rates of MHV (6.3%) and Sendai v. (16.8%) were significantly low compared with those in experimental mouse colonies. Positivity for parasites was rather high and they were infested together with other pathogens in many cases. Thus parasites including G. muris, S. muris and S. obvelata were regarded as useful indicators to see microbiological contaminations in conventional mice. There observed no strain difference in susceptibility to pathogens except for C57BL/6 and AKR mice which seemed to be high antibody responders to MHV.

Animals↗

Inactivation of Bacillus piliformis spores by heat and certain chemical disinfectants.

The inactivation of Tyzzer's organism (Bacillus piliformis) spore isolated from rats by heat and various chemical disinfectants was studied. The spores were from B. piliformis-infected rat liver tissues. The spore suspension (10(4) 50% of rat liver lesion producing dose with prednisolone treatment/ml) was treated with heart or disinfectants. Inactivation of the spores was examined in experimentally infected rats. Rats were inoculated perorally with a treated spore suspension and injected subcutaneously with prednisolone. On the sixth day after inoculation, rats were examined grossly for liver lesions. Spores were inactivated at 80 degrees C for 15 min but not at 60 degrees C for 30 min. Spores were inactivated by 0.4% peracetic acid, 0.015% sodium hypochrolite, 1% iodophol, 5% phenol. Alcide and 0.37% formaldehyde solution, but not by 0.037% formaldehyde solution, 70% ethanol, 0.3% benzethonium chloride solution, 3% cresol and soap solution, or 4% chlorhexidine digluconate. These findings suggest that B. piliformis spores are relatively sensitive to heat and certain chemical disinfectants.

Animals↗

Naturally occurring CAR bacillus infection in a laboratory rat colony and epizootiological observations.

An epizootic of chronic respiratory disease was found in a rat colony. Lungs of the symptomatic rats showed histopathologically severe peribronchial lymphoid cuffing. Filamentous bacteria were detected on the border of the tracheal and bronchial epithelium by light and electron microscopy. These bacteria did not grown on artificial media but propagated in embryonated chicken eggs. The disease was thus diagnosed as cilia-associated respiratory (CAR) bacillus infection. Epizootiological observations of the natural and experimentally induced cases revealed that the disease was highly contagious, slowly progressive and intractable. Contact infection may play a major role in the transmission of this disease.

Animals↗

[An outbreak of Trichophyton mentagrophytes infection in a rat breeding stock and its successful control].

An outbreak of dermatophytosis by Trichophyton mentagrophytes infection was observed in a breeding stock of the Wistar-Imamichi rat. Typical cutaneous lesions such as alopecia and hyperkeratosis due to the fungus were observed in 10% and 44% of adult females, and males respectively. No infection was seen in infant, even in those fostered by infected females. By 3 repeated cultivations of the hair, T. mentagrophytes was isolated from 107 (90.7%) of 118 rats clinically diagnosed as dermatophytosis. In order to maintain the original stock, the usual wholesale slaughter program was avoided and the following control measures were taken. After sacrificing all rats with the cutaneous lesions, the breeding colony was transferred temporarily to a separate animal room. The resulting vacant room was then disinfected 3 times with 10% formalin, followed by spraying with 3% sodium propionate solution 3 month later. The progenies from the non-infected mothers were dipped into 3% sodium propionate solution and transferred back into the disinfected animal room for re-colonization. Frequent sterilization of all equipment with iodophor, balsam and formalin was carried out. The room sterilized monthly with iodophor and 70% alcohol. Daily observation of all rats and monthly screening by culture test was done thereafter. As a result, no infection has been detected 4 month after recolonization, except for the 3 rats which were sacrificed immediately after detection. On the basis of the culture test and clinical observation, no infection was observed in the treated rat colony for the next five following years. Therefore, the effectiveness of these procedures of the eradication of T. mentagrophytes infection from a rat colony was shown to be quite reliable.

Animals↗

[A comparison of two antigen strains of each of mouse hepatitis virus and mouse adenovirus for detection of complement fixation antibody in mouse and rat sera].

Detection rates of complement fixation antibodies in mice and rats were compared between two antigen strains of each of mouse hepatitis virus (MHV) and mouse adenovirus (MAV). Among 66 and 47 naturally infected MHV-positive sera of mice (18 facilities) and rats (16 facilities) respectively, 17 mouse and 21 rat sera reacted with both Nu-67 and MHV-2 strains, but 49 mouse and 25 rat sera were positive to Nu-67 strain alone. Only one rat serum reacted with MHV-2 strain alone. In comparison with K87 and FL strains of MAV, all 8 positive mouse sera (3 facilities) reacted with K87 strain alone whereas out of 53 positive rat sera (20 facilities), 43, 6 and 4 sera reacted with K87 strain alone, with FL strain alone and with both the two strains, respectively.

Adenoviridae↗