The early pathogenesis in chicken inoculated with non-pathogenic serotype 2 Marek's disease virus.

A newly cloned serotype 2 Marek's disease virus (MDV), strain ML-6, was inoculated via the nasal cavity in specific-pathogen-free chicks to examine early virus replication and the expression of Marek's disease (MD)-related antigens. Following inoculation, viral intracellular antigens (VIAs) were detected in lymphoid organs (bursas and spleens) between 5 and 14 days post inoculation (PI), in feather follicles between 14 and 30 days PI, and in lungs at 3 days PI by the immunohistopathological staining of avidin-biotin-peroxidase complex method. But, very few VIAs were expressed in the thymuses between 5 and 14 days PI. However, MD tumor-associated surface antigens were not detected in any organs. Viruses were isolated from separated spleen cells at 14 and 30 days PI. Fluorescent antibodies of convalescent sera were also detected after 10 days PI. As most of the VIAs were detectable in B-cells in bursas and spleens. B-cells were considered to be the main first target cells for the serotype 2 MDV infection.     

Distribution of minke whales in the Bellingshausen and Amundsen Seas (60°W–120°W), with special reference to environmental/physiographic variables

The relationship between the distribution of minke whales ( Balaenoptera acutorostrata ) in the Bellingshausen and Amundsen Seas (longitude between 60°W and 120°W), and environmental and physiographic variables (sea-surface temperature, sea-ice extension, and sea-floor-slope type), was studied to determine whether these environmental and physical factors affect the distribution and density of minke whales. The analysis was based on sightings data obtained from the 1989/90 and 1982/83 IWC/IDCR cruises. The mean sea-surface temperatures for comparable areas were significantly higher in 1989/90 (2.04°C) than in 1982/83 (1.12°C), and the area where the sea-surface temperature was greater than 1°C in the 1989/90 study was approximately twice that of the 1982/83 study. Additionally, during the surveys, the extent of the sea ice in 1989/90 was less than that in 1982/83, with the mean ice edge about 92.6 km (50 nautical miles; 1 nautical mile ≈1.852 km) farther south in 1989/90 than in 1982/83. This is consistent with the sea ice extent observed in winter, when the sea ice extent was less in 1989 than in 1982. The distribution of minke whales was substantially different between the two surveys, with the density and abundance of minke whales being greater in 1982/83 than in 1989/90. The warmer sea-surface temperatures, fewer cold-water intrusions, and the smaller extent of sea ice in 1989/90 may be related to the difference in distribution of minke whales from 1982/83, possibly owing to the shift in availability of prey.     

Iridal coloboma induces dyscoria during miosis in FLS mice

Objective Fatty liver Shionogi (FLS) mice exhibit characteristic retinochoroidal coloboma because of a failure in fusion of the embryonic optic fissure. However, the same pathogenesis should result in iridal coloboma that has not been reported in this strain. The purpose of this study was to describe the physiologic and morphometric changes in iridal tissue involved in ocular coloboma in FLS mice. Procedures The miotic response after light exposure was evaluated in three strains of live mice, and the shape and location of the pupil were judged macroscopically. Subsequently, macroscopic abnormalities in the anterior segment and fundus were observed postmortem in all mice. During miotic and mydriatic responses in the eyes of live male FLS mice with dyscoric and normal pupils, each iris was measured in four radial directions. The enucleated eyes were examined morphometrically and histologically in both sexes of FLS mice. Results Inferior corectopia upon light‐induced miosis was clearly detected in live FLS mice. The deviated pupils were not round but oval‐shaped. Clinical and postmortem examination revealed that all dyscoric eyes had hypoplastic and dysfunctional irides inferiorly in FLS mice. Histopathological examination confirmed that both the dilator and sphincter muscles and iris stroma were quantitatively diminished in the affected inferior iris. Meanwhile, the rate of fundus (retinochoroidal) coloboma in eyes exhibiting dyscoria was remarkably high, although some dyscoric eyes had no fundus coloboma. Conclusions Fatty liver Shionogi mice had iridal coloboma, resulting in inferior corectopia upon light‐induced miosis as an indicator of ocular coloboma.     

An electron microscopic examination of fulvic acid

An electron microscopic examination of fulvic acid shows that the crystallinity, shapes, dimensions and extent of aggregation vary with pH. At pH 2.5, three types of particles can be observed: small spheroids (15–20 Å in diameter), aggregates of spheroids (200–300 Å in diameter) and an amorphous material of low contrast, perforated by voids, 500–1,100 Å in diameter. spheroidal aggregates tend to form elongated, irregularly shaped structures, 20,000–30,000 Å long.At pH 3.5, which is the natural pH of a dilute fulvic acid solution, electron micrographs show a sponge-like structure of variable thickness (100–300 Å), punctured by voids, 200–1,000 Å in diameter.At pH 4.5 and higher, electron micrographs show flat sheet-like lamellae of very low contrast, perforated by voids, 200–2,000 Å in diameter.Electron diffraction patterns show crystallinity in fulvic acid aggregates formed at pH 2.5 only but not in those formed at pH 3.5. The electron microscopic data are in harmony with X-ray, chemical and spectroscopic data on the same fulvic acid, and point to a relatively “openlrd structure, perforated by voids of varying dimensions which can trap or fix organic and inorganic compounds that fit into the voids, provided that the charges are complementary.     

Analysis of neutral glycosphingolipids from Trypanosoma brucei

Neutral glycosphingolipids (GSLs) were isolated from Trypanosoma brucei and analyzed by thin-layer chromatography (TLC), TLC/secondary ion mass spectrometry (TLC/SIMS), and liposome immune lysis assay (LILA). Three species of neutral GSLs, designated as N-1, -2, and -3 were separated on TLC. N-1 GSL migrated very close to glucosylceramide (GlcCer) and N-2 GSL showed the same mobility as lactosylceramide (LacCer). On the other hand, the mobility of N-3 GSL on the TLC plate was slower than globotetraosylceramide (Gb4). In order to characterize the molecular species of neutral GSLs from T. brucei, N-1, -2 and -3 GSLs were analyzed by TLC/SIMS. The TLC/SIMS analysis of N-1 of the parasites revealed a series of (M–H)⁻ ions from m/z 698 to 825 representing the molecular mass range of ceramide monohexoside (CMH) (GlcCer or galactosylceramide). On the other hand, the TLC/SIMS spectra of N-2 GSL revealed a series of (M–H)⁻ ions from m/z 944–987 indicating the molecular mass range of LacCer. In the TLC/SIMS analysis of N-3 GSL, however, the characteristic molecular ions that can elucidate the structure of N-3 GSL were not obtained. In order to confirm the results obtained from TLC/SIMS, N-1, -2, and -3, GSLs were tested by LILA with specific antibodies against GlcCer, LacCer, and Gb4, respectively. N-1 GSL had reactivity to anti-GlcCer antibody and N-2 GSL reacted with the antibody against LacCer. However, N-3 GSL was not recognized by anti-Gb4 antibody. Using anti-GlcCer and anti-LacCer antibodies, furthermore, we studied the expression of GlcCer and LacCer in T. brucei parasites. Both GlcCer and LacCer were detected on the cell surface of T. brucei.     

Prevalence and characterization of Staphylococcus aureus and enterotoxigenic Staphylococcus aureus in retail raw chicken meat throughout Japan

A total of 444 samples of raw chicken meat (thighs, breasts, wings, livers, gizzards, hearts and ovaries) that retailed at 145 different supermarkets in 47 prefectures in Japan were examined for contamination with Staphylococcus aureus in association with its enterotoxigenicity. S. aureus was isolated from 292 (65.8%) of the samples, and from 131 of the 145 supermarkets. There was no significant difference in the detection rate of S. aureus according to the type of meat examined. About 80% of 714 isolates belonged to the poultry (57.1%) and human biotypes (22.1%). Seventy-eight (21.7%) of 360 isolates were enterotoxigenic and isolated from 78 samples in 53 supermarkets in 31 prefectures. Staphylococcal enterotoxins (SEs) produced were SEB (50 isolates), SEA (14), SEC (8), SED (2), SEA+SEB (2), and SEA+SEC (2). Most of the enterotoxigenic isolates belonged to the human and poultry biotypes, coagulase type VII, VIII or IV, and were lysed by phages of group III. Identical SE types, biotypes, coagulase types and pulsed-field gel electrophoresis (PFGE) patterns were shown in isolates from different types of meat at the same supermarket and from samples taken from different supermarkets in the same prefectures or in isolates from samples obtained from several different prefectures. Among the 50 SEB-producing isolates, 27 yielded three similar PFGE patterns that differed by only a few fragments, suggesting that they were closely related genetically. The three patterns were found in isolates of samples that retailed at 17 supermarkets in 11 prefectures, indicating that they may be disseminated among raw chicken meat in Japan.     

Induction of immune responses against glycosphingolipid antigens: comparison of antibody responses in mice immunized with antigen associated with liposomes prepared from various phospholipids

The immune responses of mice against glycosphingolipid (GSL) antigens and the effect of the phospholipid composition of liposomes on the immunogenicity in mice of liposome-associated GSL antigens were examined. The immunization with GSL antigen alone was unable to induce any detectable anti-GSL antibody responses. On the other hand, the immune responses against GSL antigens were detected after immunization with liposomes composed of dipalmitoylphosphatidylcholine (DPPC) (0.5 micromol), cholesterol (Chol) (0.5 micromol), Salmonella minnesota R595 lipopolysaccharides (LPS) (10 microg) and GSL (0.05 micromol) (DPPC-liposome). However, the administration with liposome composed of dimyristoylphosphatidylcholine (DMPC) (0.5 micromol), Chol (0.5 micromol), S. minnesota R595 LPS (10 microg) and GSL (0.05 micromol) and with liposomes composed of distearylphosphatidylcholine (DSPC) (0.5 micromol), Chol (0.5 micromol), and S. minnesota R595 LPS (10 microg) and GSL (0.05 micromol) was ineffective for the induction of the immune responses against GSL antigens. These results suggest that DPPC-liposome would serve effectively as a delivery vehicle for inducing immune responses against GSL antigen.     

Detection of fungi producing infection-inhibiting metabolites against <Emphasis Type="Italic">Alternaria alternata</Emphasis> Japanese pear pathotype from fungi inhabiting internal tissues of Japanese pear shoots

Fungi inhabiting Japanese pear were isolated from internal tissues of cv. Nijisseiki, and culture filtrates (CFs) of 100 isolates were evaluated for their inhibitory activity against infection by Alternaria alternata Japanese pear pathotype. CFs of 11 isolates inhibited lesion formation on the pear by the pathogen. Among these isolates, CFs of five isolates inhibited spore germination. CFs of the six other isolates inhibited appressorial formation, infection hypha formation, AK-toxin production, or a combination of these actions. Analysis of sequence homology in the rDNA ITS1 regions of these isolates showed that most isolates had high homology with some fungal endophytes.     

Identification of glycosphingolipid binding sites for SEF21-fimbriated Salmonella enterica serovar Enteritidis in chicken oviductal mucosa

In order to clarify the presence of glycosphingolipids (GSLs) receptors for Salmonella enterica serovar Enteritidis with SEF21 fimbriae, we analyzed neutral GSLs and gangliosides from chicken oviductal mucosa and investigated the binding of bacteria to neutral GSLs and gangliosides. Five types of neutral GSLs, designated as N-1 to N-5, and two types of gangliosides, designated as G-1 and G-2, were identified on the thin-layer chromatography (TLC) plates. In the bacterial binding assay on TLC, the fimbriated bacteria bound only to glucosylceramide (GlcCer) standard, N-1 having the same TLC mobility as GlcCer, GM3 standard and G-1 corresponding to GM3 in TLC mobility, but not to N-2, N-3, N-4, N-5, or G-2. These results suggest the presence of GlcCer (N-1) and ganglioside GM3 (G-1) on the epithelial surface of chicken oviductal tract which act as sites for adherence of SEF21-fimbriated S. Enteritidis.