Evaluation of PCR as a means of identification of Pasteurella pneumotropica.
A polymerase chain reaction with new primers (new PCR) designed from Pasteurella pneumotropica 16S rDNA as an identification system for this organism was compared with the PCR reported by Wang et al. (Wang's PCR) by using 15 bacterial reference species and 70 clinical isolates with the conventional identification system. For the 15 reference strains, both PCRs were identical. For the 70 clinical isolates, the new PCR and Wang's PCR showed consistency with the conventional system in 62.9% (44/70) and 51.4% (36/70), respectively. Twenty-six isolates were inconsistent with the conventional system and the new PCR with respect to morphology and serology. These findings suggested that the new PCR was more sensitive than Wang's PCR, and the new PCR in combination with morphology and serology is useful for P. pneumotropica identification. (+info
Opportunistic Pneumocystis carinii infection in red-bellied tamarins (Saguinus labiatus).
P. carinii infection in red-bellied tamarins (Saguinus labiatus), born and maintained in a laboratory breeding colony, was examined by histopathologic examination postmortem. P. carinii cysts were detected in 6 of 10 red-bellied tamarins examined, by using Grocott's, toluidine blue O and immunostaining with avidin-biotin complex using antisera for rat-, simian-, and human-P. carinii. The results obtained from the present studies imply that P. carinii may be an important pathogen in this species. (+info
Microsatellite loci in wild-type and inbred Strongylocentrotus purpuratus.
Strongylocentrotus purpuratus, a major research model in developmental molecular biology, has been inbred through six generations of sibling matings. Though viability initially decreased, as described earlier, the inbred line now consists of healthy, fertile animals. These are intended to serve as a genomic resource in which the level of polymorphism is decreased with respect to wild S. purpuratus. To genotype the inbred animals eight simple sequence genomic repeats were isolated, in context, and PCR primers were generated against the flanking single-copy sequences. Distribution and polymorphism of these regions of the genome were studied in the genomes of 27 wild individuals and in a sample of the inbred animals at F2 and F3 generations. All eight regions were polymorphic, though to different extents, and their homozygosity was increased by inbreeding as expected. The eight markers suffice to identify unambiguously the cellular DNA of any wild or F3 S. purpuratus individual. (+info
Infection of laboratory mice with the human granulocytic ehrlichiosis agent does not induce antibodies to diagnostically significant Borrelia burgdorferi antigens.
Laboratory diagnosis of Borrelia burgdorferi is routinely made by an enzyme-linked immunosorbent assay, with positive results confirmed by Western blot analysis. Concern has been raised that false-positive diagnoses may be made on the basis of serologic cross-reactivity with antibodies directed against other bacterial pathogens, in particular the agent of human granulocytic ehrlichiosis (HGE). The present study made use of a mouse model to ascertain the validity of these concerns. Two different strains of mice were inoculated with the HGE agent and assayed for production of polyclonal and monoclonal antibodies to antigens of both of these bacteria. Infection of mice with the HGE agent does not induce diagnostically significant B. burgdorferi serologic cross-reactions. (+info
Genetics of mouse behavior: interactions with laboratory environment.
Strains of mice that show characteristic patterns of behavior are critical for research in neurobehavioral genetics. Possible confounding influences of the laboratory environment were studied in several inbred strains and one null mutant by simultaneous testing in three laboratories on a battery of six behaviors. Apparatus, test protocols, and many environmental variables were rigorously equated. Strains differed markedly in all behaviors, and despite standardization, there were systematic differences in behavior across labs. For some tests, the magnitude of genetic differences depended upon the specific testing lab. Thus, experiments characterizing mutants may yield results that are idiosyncratic to a particular laboratory. (+info
Production of ex-germfree rabbits for establishment of specific pathogen-free (SPF) colonies.
Nine groups of ex-germfree (GF) rabbits were produced by inoculation of hysterectomy-derived GF rabbits with various combinations of cecal bacteria isolated from conventional (CV) rabbits in order to establish a barrier-sustained colony. Six strains of Bacteroides and two strains of Streptococcus isolated from CV rabbits (2 to 3 weeks old) were used for pretreatment. The mortality of ex-GF rabbits inoculated with the anaerobic growth (CF) on EG or SM10 plates inoculated with a 10(-5) dilution of cecal contents was 71.4 to 94.4% when given without pretreatment. All ex-GF rabbits pretreated with Bacteroides alone survived, but the mortality of ex-GF rabbits inoculated with Bacteroides plus Streptococcus strains as pretreatment was 20 and 45.4%. The mortality of ex-GF rabbits inoculated with only Bacteroides was 43%. All ex-GF rabbits inoculated with Bacteroides plus anaerobic growth (CF), cecal suspension of ex-GF mice which had been inoculated with cecal suspensions of CV rabbits (MF) or chloroform-treated cecal suspension (CHF) survived, but CHF inoculated ex-GF rabbits were in an unhealthy condition with slight diarrhoea. These data indicate that inoculation with Bacteroides strains as pretreatment plus CF or MF was required to convert GF rabbits to the normal state. (+info
Establishment of specific pathogen-free rabbit colonies with limited-flora rabbits associated with conventional rabbit flora, and monitoring of their cecal flora.
In the present study we attempted to establish specific-pathogen-free (SPF) rabbit breeding colonies with two groups of limited-flora (LF) rabbits, both ex-germfree rabbits, and their offspring. Two groups of LF rabbits associated with cecal flora of conventional (CV) rabbits produced in a previous study [Exp. Animals, submitted], were transferred to individual barrier rooms and some of the LF rabbits were accommodated in isolators to maintain the basic flora for SPF rabbits. The composition of the cecal flora of LF rabbits was stable for a long period; bacteroides remained predominant and clostridia dominant. From the SPF rabbits, different types of bacteria, e.g., enterobacteriaceae and streptococci, which could not be isolated in the isolator were detected at a low population level at an early stage in the establishment of the SPF colonies, but the basic composition of the cecal flora was mainly bacteroidaceae and clostridia and did not change over a long period, and the floral composition became similar to that of CV rabbits. The fertility and weaning rates of the SPF rabbits were satisfactory for a SPF rabbit colony. In addition, these SPF colonies were free of more than one year rabbit-specific pathogens. (+info
Leukocyte differential analysis in multiple laboratory species by a laser multi-angle polarized light scattering separation method.
Leukocyte differential analysis was performed in various species, particularly laboratory animals, by the laser multi-angle polarized light scattering separation method. Venous blood specimens were drawn from the following subjects: healthy adult men and women ("humans"); cynomolgus monkeys ("monkeys"); common marmosets ("marmosets"); beagle dogs ("dogs"); miniature potbelly pigs ("swine"); Japanese white rabbits ("rabbits"); Hartley guinea pigs ("guinea pigs"); and Sprague-Dawley rats ("rats"). 90 degrees/10 degrees scatter plot: Basophils and mononuclear-polymorphonuclear cells were separated in all subjects, but individual 10 degrees and 90 degrees scatter plots overlapped in dogs and guinea pigs, respectively. 90 degrees depolarized /90 degrees scatter plot: Neutrophils and eosinophils were clearly separated in human, monkey, guinea pig, swine and rat subjects. The eosinophil cluster was not clearly plotted in marmoset, dog, or rabbit. 0 degree/10 degrees scatter plot: Regarding this plot for monocytes and lymphocytes, cells were plotted in the following order in all subjects: lymphocytes < basophils < or = monocytes in the 0 degree (size) scatter; and lymphocytes [symbol: see text] monocytes < or = basophils in the 10 degrees (complexity) scatter. Compared to other species, the rat scatter showed a tendency to overlapping plots in both the 0 degree and 10 degrees scatters in the monocyte and lymphocyte clusters. In both dog and guinea pig, the monocyte and neutrophil plots overlapped in the 0 degree and 10 degrees scatters. Basobox: In the human and rabbit subjects, the basophil cluster was plotted within the established basobox, but no clear cell cluster was plotted in the other subjects. As a result of comparing the percentage values for leukocytes in various species obtained by using the CD3500 apparatus versus the corresponding values obtained manually, good correspondence was found in the monkey, and eosinophils in the marmoset were lower with CD3500 than manually. In the rabbit, the mean measured value for basophils matched in the manual and CD3500 findings. In the guinea pig, the CD3500 values were lower than the manual values for lymphocytes, but higher for monocytes and neutrophils. The above findings suggest that the laser multi-angle polarized light scattering separation method is indeed capable of analyzing leukocytes from various species based on cell size and cell complexity, i.e., the presence or absence of nuclei, granules and cell enclosures. (+info