Seed formation in triploid loquat (Eriobotrya japonica) through cross-hybridization with pollen of diploid cultivars

As the fruits of loquat (Eriobotrya japonica, 2n = 2x = 34) carry large seeds, the breeding of seedless loquat has long been a goal. The recent creation of triploid cultivars (2n = 3x = 51) and the application of gibberellins allow commercial production of seedless loquat, but the possibility of seed formation in triploid loquats has not been carefully investigated. Through crossing experiments and cytological observations of meiosis and pollen tube growth, we found that the triploid line 3N-N28 was essentially self-sterile, but developed seeds on pollination with pollen from diploid cultivars at rates of up to 5.5%. Almost half of the seedlings survived to 5 months, and carried diploid (2n = 34), tetraploid (2n = 68), or aneuploid chromosome numbers. Our results suggest that triploid loquat cultivars might retain the risk of seed formation. Protection from pollination by diploid cultivars or the development of new triploid cultivars will be necessary to ensure the production of seedless loquat fruits.     

Influence of initial swimbladder inflation failure on survival of Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

This study investigated the influence of initial swimbladder inflation (ISI) failure on survival of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae to prevent mass mortality due to sinking death. In Experiment 1, swimbladder inflation frequency and survival within an ISI promoted (PS) group, for which surface film of rearing water was removed, and a group without ISI promotion (NPS) were compared in 20 and 30 kL tanks. The PS group demonstrated significantly higher swimbladder inflation frequency and increased survival than the NPS group within 20 kL tanks at 9 days post hatching (dph). Similar tendencies were observed within 30 kL tanks. In Experiment 2, larval distribution and swimbladder inflation frequency in the night‐time were examined through larval sampling within the upper and middle layers of the water column and tank bottom within 1.6 and 30 kL tanks at 5 dph. Larvae at tank bottom had higher distributional density and significantly lower swimbladder inflation frequency than those distributed in the upper and middle layers within 1.6 kL tanks. Similar tendencies were observed within 30 kL tanks. Results of this study suggest the improvement of larval survival due to prevention of sinking death through ISI promotion in mass‐scale PBT larviculture.     

Cyanobacterial communities of rice straw left on the soil surface of a paddy field

To estimate diversity, seasonal variation, and phylogeny of the cyanobacterial communities in rice straw placed in nylon mesh bags and left on the soil surface of a paddy field, total DNA was extracted from straw, amplified by polymerase chain reaction targeting 16S rRNA genes of cyanobacteria, and the amplicons were separated by denaturing gradient gel electrophoresis (DGGE). These DGGE bands were sequenced. The paddy field was under flooded condition after transplanting of rice (Experiment 1) and under drained conditions after harvest (Experiment 2). The residual samples on the soil surface under upland conditions were collected just before spring plowing and were placed again on the soil surface after transplanting under flooded conditions. DGGE band patterns of cyanobacterial communities of rice straw were different under drained conditions, under flooded conditions when fresh rice straw samples were placed (Experiment 1), and under flooded conditions when residual rice straw samples were replaced (Experiment 2), indicating that the communities were influenced by both water regime of the paddy field and the degree of the rice straw decomposition. Sequence analysis of DGGE bands indicated that most of the cyanobacteria in rice straw on the soil surface in the paddy field were filamentous members belonging to Subsections III and IV. Filamentous cyanobacterial cells were observed in rice straw under flooded conditions by epifluorescence microscopy.     

An angiotensin converting enzyme inhibitor,benazepril can be transformed to an active metabolite,benazeprilat, by the liver of dogs with ascitic pulmonary heartworm disease

To examine whether an angiotensin converting enzyme (ACE) inhibitor, benazepril, can be transformed to the active metabolite, benazeprilat, by severely injured liver of dogs with ascitic heartworm disease, benazepril hydrochloride was administered orally to dogs once daily for 7 consecutive days at a dose rate of 0.29 mg/kg to 0.63 mg/kg of body weight, and plasma benazepril and benazeprilat concentrations were determined on the 1st and 7th administration days. In 7 dogs with ascitic pulmonary heartworm disease, plasma benazeprilat concentrations tended to be higher than in 7 control dogs both on the 1st and 7th administration days. The peak concentration and area under the concentration-time curve tended to be greater in dogs of the ascites group than in control dogs, but the statistics could not detect significant differences in the time to peak concentration and t(1/2) between the control and ascites groups. Plasma ACE activities decreased after administration of benazepril. In dogs with ascitic heartworm disease, benazepril was readily transformed to benazeprilat by the liver, and was effective for suppression of plasma ACE activity.     

Latency and persistence of bovine herpesvirus type 4, strain B11-41, in bovine nervous tissues

Three cattle were experimentally infected with bovine herpesvirus type 4 (BoHV-4), strain B11-41, isolated from the spinal cord of a cow, and monitored for clinical symptoms. None of them showed any clinical signs except increases of leukocyte numbers in two of them, and the body temperature remained normal throughout the experiment. Antibody titers against BoHV-4 continuously increased for one month and were maintained at a high level for more than 1 year by enzyme-linked immunosorbent assay (ELISA). The virus was isolated only from serum and peripheral blood leukocytes (PBL) of one cow in the early stage of infection, but the viral genome was detected in PBL continuously by PCR. When they were euthanized, the viral genome was detected in the lymph nodes and nervous tissues such as medulla, spinal cord, and trigeminal ganglion. These results indicate that cattle are infected with the virus latently and persistently, and the latency site would be in the tissues of the central nervous system as well as lymphoid tissues. When a seroepidemiological survey was performed on antibodies to BoHV-4 among cattle in Japan by ELISA, the rate of antibody-positive cattle was 8.9% and they were found irregularly on certain farms.     

Gummy stem blight of balsam pear caused by <Emphasis Type="Italic">Didymella bryoniae</Emphasis> and its anamorph <Emphasis Type="Italic">Phoma cucurbitacearum</Emphasis>

Gummy stem blight of balsam pear found in the Kanto district and in the Hokkaido Prefecture was demonstrated to be caused by Didymella bryoniae (Auerswald) Rehm based on inoculation experiments, molecular analysis, and morphological identification of the pathogenic fungus. This fungus was also pathogenic to related plants belonging to Cucurbitaceae. The imperfect stage of the fungus was identified as Phoma cucurbitacearum (Fr.: Fr.) Sacc. based on morphological similarities.     

Volatile organic compounds from wood and their influences on museum artifact materials I. Differences in wood species and analyses of causal substances of deterioration

Wood is generally used as the interior material in museum storage rooms. Recently, however, the effects of volatile organic compounds (VOCs) from wood on artifacts has become a topic of great concern. The VOCs from four species of wood (western red cedar, spruce, kiri, and sugi) and their effects on artifact materials (two types of metal, seven types of pigment) were investigated using a deterioration-accelerating test, gas chromatography-mass spectrometry, X-ray diffraction analysis, and X-ray photoelectron spectroscopy. The results suggested that the influences on artifact materials varied greatly with wood species, and depended on specific components such as hinokitiol or acetic acid rather than the amount of total volatile organic compounds (TVOCs). It is a very serious problem that of the four species of wood, western red cedar (rich in hinokitiol), which has been recommended as an interior material for museum storage rooms, showed the heaviest deterioration on metal samples, and only this type of wood discolored enpaku (white lead) and rokushou (malachite, verdigris). In such storage rooms, museum artifacts should be carefully monitored. When selecting an interior material for a storage room or studying methods of preventing deterioration, it is very important to consider fully the characteristics of wood VOCs, not only the amount of TVOC.Part of this paper was presented at the 24th (Tokyo, June 2002) and 25th (Kyoto, June 2003) Annual Meeting of the Japan Society for the Conservation of Cultural Property     

Evaluation of the live vaccine efficacy of virulence plasmid-cured,and phoP- or aroA-deficient Salmonella enterica serovar Typhimurium in mice

We evaluated the protective efficacy of 94-kb virulence plasmid-cured, and phoP- or aroA-deficient strains of Salmonella enterica serovar Typhimurium (ΔphoP or ΔaroA S. Typhimurium) as oral vaccine candidates in BALB/c mice. Two weeks after the completion of 3 oral immunizations with 1 × 10⁸ colony-forming units (CFU) of virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium at 10-day intervals, S. Typhimurium lipopolysaccharide (LPS)-specific mucosal secretory immunoglobulin A (s-IgA) antibody titers were detected in the cecal homogenate, bile and lung lavage fluid, but not in the intestinal lavage fluid. In addition, the increases in S. Typhimurium LPS-specific immunoglobulin G (IgG) and IgA antibody titers in the serum were also observed 2 weeks after completing 3 oral immunizations with virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium. The series of 3 oral immunizations protected the mice against an oral challenge with 5 × 10⁸ CFU of the virulent strain of S. Typhimurium, suggesting that both the virulence plasmid-cured, and ΔphoP and ΔaroA S. Typhimurium strains are promising candidates for safe and effective live S. Typhimurium vaccines.     

Measurement of porcine luteinizing hormone concentration in blood by time-resolved fluoroimmunoassay

We validated a time-resolved fluoroimmunoassay (TR-FIA) using the lanthanide element europium as a non-radiological tracer for measuring luteinizing hormone (LH) in porcine peripheral blood. The dose-response curve of the reference standard ranged from 0.2048 to 50 ng/ml. Good parallelism was noted between the LH standard and plasma sample. The profile of LH throughout the estrous cycle, assessed by daily blood sampling, was consistent with the previous findings obtained by radioimmunoassays (RIAs). Moreover, the secretory patterns of pulsatile LH in the follicular phase and the preovulatory LH surge were also similar to those obtained in previous RIAs. We conclude that TR-FIA can be used to measure LH levels in porcine blood, with practical and convenient applications.     

Japanese black cattle with orotic aciduria detected by gas-chromatography/mass-spectrometry

A 4-months-old calf of Japanese black cattle was diagnosed with orotic aciduria by gas-chromatography/mass-spectrometry (GC/MS). Until now orotic aciduria had not been reported in Japanese black cattle. The animal showed repeated diarrhea. The hematocrit was low, and microcytes and acanthocytes were observed in blood smears. The calf had lower serum total protein concentrations with a higher blood ammonia concentration. Needle-shaped crystals of orotic acid were observed in urinary sediments. Sequence homologous analysis with cattle uridine monophosphate synthase DNA indicated silent mutation in the affected calf.