Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen.

Forty peptides were synthesized corresponding to hydrophilic clusters of amino acids within the sequences of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP). Six peptides were studied in more detail and the most promising, a 2B peptide, was evaluated in enzyme-linked immunosorbent assay (ELISA) using sera from naive, vaccinated, and vaccinated-and-challenged cattle as well as bovine sera from field outbreaks. The performance of the new NSP peptide ELISA was compared to that of 4 commercial NSP ELISA kits. Antibody to 2B was detectable from the end of the first week to the second week after infection in most of the nonvaccinated animals and by the second to third week in vaccinated-and-challenged animals. The sensitivity of the 2B peptide ELISA was comparable to the 3ABC Ceditest (Ceditest FMDV-NS, Cedi Diagnostics B.V.; Chung et al., 2002). With some modification and further validation, this 2B test could be useful as a screening or conformational NSP test in postvaccination surveillance for FMD.     

Oxygen isotopic compositions of asteroidal materials returned from Itokawa by the Hayabusa mission

Meteorite studies suggest that each solar system object has a unique oxygen isotopic composition. Chondrites, the most primitive of meteorites, have been believed to be derived from asteroids, but oxygen isotopic compositions of asteroids themselves have not been established. We measured, using secondary ion mass spectrometry, oxygen isotopic compositions of rock particles from asteroid 25143 Itokawa returned by the Hayabusa spacecraft. Compositions of the particles are depleted in (16)O relative to terrestrial materials and indicate that Itokawa, an S-type asteroid, is one of the sources of the LL or L group of equilibrated ordinary chondrites. This is a direct oxygen-isotope link between chondrites and their parent asteroid.     

Polymerase chain reaction-restriction fragment length polymorphism analysis of the SzP gene of Streptococcus zooepidemicus isolated from the respiratory tract of horses

OBJECTIVE: To develop polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis for molecular typing of strains of Streptococcus zooepidemicus and to use the new typing method to analyze a collection of isolates from the respiratory tract of Thoroughbreds. SAMPLE POPULATION: 10 strains of S zooepidemicus, 65 isolates from the respiratory tract of 9 yearlings following long distance transportation, and 89 isolates from tracheal aspirates of 20 foals with pneumonia. PROCEDURE: Phenotypic variations in the SzP protein were detected by western immunoblot analysis. Using PCR-RFLP analysis, genotypes were obtained with primer sets from the SzP gene, followed by restriction endonuclease digestion of the amplicons. RESULTS: Unique genotypic patterns were obtained with a primer set designed from both ends of the structural gene and the restriction endonuclease DdeI. Forty-five isolates from the lymphoid tissue within the pharyngeal recess (ie, pharyngeal tonsil) of yearlings included 10 SzP genotypes and SzP phenotypes. Isolates from the trachea of each yearling were of a single genotype that was also present among isolates from the pharyngeal tonsil of the same horses. Isolates from tracheal aspirates of foals belonged to 14 genotypes. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of the SzP gene by use of PCR-RFLP was effective for molecular typing of strains of S zooepidemicus in the study of respiratory tract disease in horses. Results of PCR-RFLP analysis indicate that a single strain of S zooepidemicus can migrate from the pharyngeal tonsil to the trachea at a high rate in horses undergoing long distance transportation.     

Histocytological specificities of adrenal cortex in the New World Monkeys, Aotus lemurinus and Saimiri boliviensis

The New World monkey Aotus spp. (night monkeys) are expected for use of valuable experimental animal with the close species of Saimiri spp. (squirrel monkeys). Saimiri is known to show spontaneous hypercortisolemia, although few reports in Aotus. We compared basic states of blood steroid hormones and histological structure of the adrenal glands in two monkeys. Serum cortisol and ACTH levels were statistically lower in Aotus than Saimiri. Conversely, Aotus adrenocortical area showed significant enlargement, especially at the zona fasciculata. Electron microscopic observation at Aotus fasciculata cells revealed notable accumulation of large lipid droplets and irregular shapes of the mitochondrial cristae. These results suggest potential differences in cellular activities for steroidogenesis between Aotus and Saimiri and experimental usefulness in adrenocortical physiology and pathological models.     

Cycas necrotic stunt virus isolated from gladiolus plants in Japan

An undescribed spherical virus ca. 30 nm in diameter was isolated from gladiolus (Gladiolus spp.) plants in Japan. The virus had a moderate host range within eight families. Purified virus preparations contained two large RNA components and one coat protein with mobility similar to Cycas necrotic stunt virus (CNSV) from cycas (Cycas revolute). The virus was serologically closely related to CNSV. Its nucleotide sequence of the coat protein gene had 89% common identity with that of CNSV. These results indicated that the virus isolated from gladiolus is a new strain of CNSV. The nucleotide sequence data reported are available in the DDBJ/EMBL/Gen Bank databases under the accession number AB237656.     

Follicular dynamics during the pre-ovulatory and subsequent first follicular wave stages affect the pregnancy outcome in Japanese Black cows

The diameters of the pre-ovulatory follicles (PF) and the largest follicle during the subsequent first follicular wave (W1LF), and plasma estradiol-17β (E₂) concentrations were monitored on Days 0, 1, 3, 5, and 7 (ovulation = Day 1). Pregnancy was diagnosed on Day 30. Cows were classified into two groups according to the location of the dominant follicle ipsilateral (IG) or contralateral (CG) to the corpus luteum on Day 7. From Days 3 to 7, some follicles that had been determined as the subordinate in the previous examination exceeded the W1LF located in the opposite ovary in terms of the diameter. These were defined as switching (SW), whereas others were defined as non-switching (NSW). The diameter of PF was significantly smaller in pregnant (P) animals than in non-pregnant (NP) animals. The plasma E₂ concentration on Day 0 was significantly higher in P animals than in NP animals and tended to be higher in NSW than in SW. In addition, plasma E₂ concentrations around Days 3 to 7 tended to be higher in P animals of NSW than in NP animals of SW. The conception rates did not differ between IG and CG but were significantly higher in NSW than in SW. In the IG group, the conception rate tended to be higher in NSW than in SW.     

Population genetic structure of <Emphasis Type="Italic">Microdochium majus</Emphasis> and <Emphasis Type="Italic">Microdochium nivale</Emphasis> associated with Fusarium head blight of wheat in Hokkaido,Japan

The extent of mycotoxin-induced responses in living roots of two maize (Zea mays L.) cultivars differing in their susceptibility to Fusarium was studied. Application of mycotoxin zearalenone (ZEN) or its derivates α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL) caused a rapid depolarization of plasma membrane potential (E_M). The extent of E_M depolarization was dependent on the type of mycotoxin that have been used and showed concentration dependence. Interestingly, ZEN, but not its derivatives α-ZEL and β-ZEL, significantly decreased respiration of maize root cells. Electrolyte leakage increased with the duration of toxins treatment and was significantly higher in susceptible cv. Pavla than in resistant cv. Lucia. A strong superoxide dismutase insensitive nitro blue tetrazolium (NBT) reduction activity was identified in the root tips of control plants. This activity was rapidly inhibited by mycotoxin application in the meristematic/distal transition zones of roots in both cultivars examined. The level of recovery was a function of the mycotoxin concentration. Moreover, mycotoxin treatment also resulted in the onset and the progression of root cell death which was dependent on both, the type and concentration of mycotoxin.     

Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.     

Androgen induces production of male effect pheromone in female goats

Previously we showed that the primer pheromone responsible for the "male effect" was produced in specific skin regions of castrated male goats by androgen treatments. In the present study, we examined whether androgen can also induce production of the male effect pheromone in female goats. Capsules containing dihydrotestosterone (DHT) or testosterone (T) were subcutaneously implanted into six ovariectomized (OVX) goats for 28 days. Small skin samples were collected from the head and rump regions, and the pheromone activity of their ether extracts was examined using a bioassay that monitors the electrophysiological manifestation of the hypothalamic gonadotropin-releasing hormone pulse generator as multiple-unit activity. Behaviors of OVX goats towards ovary-intact estrous goats were also examined before and at the end of DHT or T treatment. Before androgen treatment, neither the head nor rump skin samples in OVX goats showed pheromone activity. DHT treatment induced pheromone activity in the head skin sample of six OVX goats and in the rump skin sample of two OVX goats. Similar results were obtained by T treatment. In addition, OVX goats treated with T showed masculine-type sexual behaviors such as courtship and mounting behaviors towards the estrous goats. These results demonstrate that androgen is capable of inducing primer pheromone activity in the female and suggest that the synthesis pathway of the male effect pheromone exists in both sexes in the goat.