Neospora caninum and complex vertebral malformation as possible causes of bovine fetal mummification

Bovine neosporosis, caused by Neospora caninum is a leading cause of abortion in cattle. We postulated that neosporosis could lead to fetal death and mummification. Fifteen mummified fetuses were tested by polymerase chain reaction (PCR) for the mutation in the bovine SLC35A3 gene that causes complex vertebral malformation (CVM) and the pNC-5 gene which identifies N. caninum infection. DNA was extracted from the mummified fetuses and the sex of the mummies was determined by PCR. The CVM mutation was not detected in the mummified fetuses, but 4 fetuses were positive for N. caninum infection. The ages of the mummies with N. caninum infection were 100, 113, 123, and 131 days. Twelve of the 15 mummified fetuses were male. To our knowledge, this is the first detection of N. caninum as a possible cause of bovine fetal mummification.     

Biodegradation of nonlignocellulosic substances II: physical and chemical properties of sawdust before and after use as artificial soil

The physical properties of sawdust including porosity, water retention, and water drainage were analyzed to prove its suitability for use as an artificial soil in the automatic decomposer-extinguisher (GADE) machine. The physical and chemical properties of residual sawdust from the GADE machine were also analyzed, the mechanical abrasion of sawdust in the GADE machine was tested, and the morphology of this residue was observed through a scanning electron microscope to investigate changes of these properties in the medium of decomposing garbage. Sawdust, which showed a lower specific gravity and larger porosity than soil, is considered capable of supplying air to bacteria. It was found that sawdust became worn from the operation of the machine. The spaces of residual sawdust were still observed, but water drainage decreased. The portion of hollocellulose in residual sawdust decreased, although the extractives in it increased. Results indicated that the capacity of sawdust to function as an artificial soil in the GADE machine was decreased owing not only to the destruction of sawdust grain but also to the adherence of products from decomposition, such that sawdust needed to be replaced every few months.Part of this report was presented at the 43rd annual meeting of the Japan Wood Research Society, Morioka, April 1993     

Screening of heterozygous DNA markers in shiitake (Lentinula edodes) using de-dikaryotization via preparation of protoplasts and isolation of four meiotic monokaryons from one basidium

A suitable screening method for heterozygous DNA markers in shiitake,Lentinula edodes (Berk.) Pegler, is reported. Monokaryons were derived from a dikaryon by de-dikaryotization via protoplast formation. Compatibility of the monokaryons was determined by pairwise culture on agar plates. We selected the primers to amplify polymorphic fragments among the original strain (Hokken600H600) and two monokaryons (H600PP-39 and H600PP-67) showing compatibility. A total of 135 fragments were selected as specific random amplified polymorphic DNAs (RAPDs) resulting from 56 primers of the 147 primers tested. Furthermore, we tested whether the polymorphic fragments segregated into 22 among four strains isolated from a basidium. Most of the polymorphic fragments (about 97.8%) showed 22 segregation among the four strains. We concluded that the polymorphic fragments were heterozygous if they were detected in either of the monokaryons (H600PP-39 and H600PP-67) and segregated to 22 among four meiotic strains (H600B-1,-2, -3, and -4). A total of 132 heterozygous DNA markers were therefore selected from a dikaryon of shiitake (Hokken600H600).Part of this report was presented at the 8th Annual Meeting of Mushroom Science and Biotechnology and the 47th Annual Meeting of the Japan Wood Research Society, Kochi, 1997     

Effects of age, egg-laying activity, and Salmonella-inoculation on the expressions of gallinacin mRNA in the vagina of the hen oviduct

Gallinacins (Gal) are antimicrobial peptides that play significant roles in innate immunity in chickens. The aim of this study was to examine whether age of birds and egg-laying activity (laying and non-laying caused by feed-regulation) affect the mRNA expression of Gal-1, -2, and -3 in the vagina of hens, and whether their expressions are changed in response to the stimulation with Salmonella enteritidis (SE) and lipopolysaccharide (LPS). White Leghorn hens were divided into groups of young and old laying hens, and groups of laying and non-laying hens after feed-regulation. Vaginal cells were cultured and stimulated with SE or LPS. Expressions of Gal-1, -2, and -3 mRNA in their vaginal mucosa and cultured cells were examined by quantitative real-time RT-PCR. The expressions of Gal-1, -2, and -3 of the vaginal mucosa were significantly greater in old birds than in young birds. Expression of these Gals in the vagina were decreased in the regressed oviduct of non-laying birds compared with laying birds. The expressions of Gal-1, -2, and -3 in the cultured vaginal cells were increased by stimulation with SE or LPS within 24 h. These results suggest that the mRNA expressions of Gal-1, -2 and -3 in the vagina of laying hens increased with age, whereas they decreased in the regressed oviduct during the non-laying phase. Also, synthesis of these antimicrobial peptides in the vagina may increase in response to SE and LPS to eliminate SE bacteria.     

Storage of maitake mushroom (Grifola frondosa) culture medium after harvesting fruit bodies is an effective pretreatment for ethanol conversion

The storage of spent maitake culture medium (SMCM) under various conditions was investigated as a potential pretreatment of SMCM for increased ethanol conversion. When SMCM was stored at 25°C for 12?weeks, the glucose yield by enzymatic saccharification increased from 22 to 52%. Selective degradation of lignin and hemicellulose occurred in SMCM during storage. The optimal storage temperature of SMCM was at the active growing temperature of maitake mycelium, which is between 25 and 30°C. Storage of SMCM under anaerobic conditions did not increase the glucose yield compared to non-stored conditions. The glucose yield from the SMCM stored for 4?weeks at 25°C was increased by about 30% with either NaOH or vibrating ball milling pretreatment. After 12?weeks of storage, the glucose yield from SMCM without any other pretreatment was higher than that of non-stored SMCM with additional pretreatments. An ethanol yield of 42.1% was obtained from SMCM stored for 12?weeks by simultaneous saccharification and fermentation, which was comparable to yields after NaOH (41.3%) or vibrating ball milling (44.1%) pretreatments. Therefore, storage of SMCM is a very useful pretreatment for bioethanol production.     

Characteristics in Concentration of Chemical Species in Ambient Air Based on Three-Year Monitoring by Filter Pack Method

Continuous monitoring of concentrations of particulate matter and gaseous compounds for three years by a four-stage filter-pack method clarified the characteristics in the concentrations of chemical species in particulate matter and gaseous compounds in ambient air in Kobe, Japan. The amount of materials in blank filters was low enough for this method to be used for the monitoring of ambient air. Little or none of the hydrogen chloride was derived anthropogenically. The concentrations of both particulate sulfate and sulfur dioxide in spring and summer were significantly higher than those in autumn and winter. The concentration of gaseous nitric acid was high in summer, which suggests that active photochemical reactions in the atmosphere provided more nitrogen species in summer. The measured concentration products [HNO₃][NH₃] showed good agreement with the theoretical predictions with some exceptions. The total concentration of ammonium species showed no significant seasonal variation except that the concentration in winter was low. The concentration of nss-Ca²⁺ in spring was significantly higher than that in the other seasons, which is most likely associated with the so-called yellow-sand events. The concentrations of nss-K⁺ and nss-Ca²⁺ accounted for most of K⁺ and Ca²⁺, respectively, whereas that of nss-Mg²⁺ accounted for only 5% of Mg²⁺     

Complete nucleotide sequence of mitochondrial DNA in White Leghorn and White Plymouth Rock chickens

Among the chicken breeds, White Leghorn (WL) and White Plymouth Rock (WR) are major breeds and have different history in their establishments. Whole mitochondrial DNA of the breeds were sequenced in order to elucidate the genetic relationship between the breeds. The lengths of the two WL and two WR mitochondrial DNA were found to be 16 788 and 16 785 base pairs, respectively. When the DNA sequences were compared, the similarity was found to be 99.96% (six nucleotide differences). In addition, the present study conformed the existence of an extra nucleotide ‘C’ in NADH dehydrogenase subunit 3 (ND3) of the chicken mitochondrial DNA, which has been consistently observed in Galliformes.     

Cucumber mosaic virus isolated from Aconitum spp. in Japan

Viruses were isolated from leaves of plants of Aconitum species with symptoms such as mottling and yellowing in Hokkaido and Gunma prefectures in Japan. These viruses were identified as Cucumber mosaic virus (subgroup II) based on particle morphology, host range, aphid transmission, and serology.     

Sperm motility, plasma membrane integrity, and binding capacity to homologous zona pellucida of cryopreserved epididymal spermatozoa in the domestic cat

We examined motility, plasma membrane integrity, and binding capacity to homologous zona pellucidae (ZP) of frozen/thawed epididymal cat sperm as a model species for endangered felines. Epididymal spermatozoa from 20 domestic cats were frozen with freezing egg-yolk extender containing 3.0% glycerol in 0.25-ml straws. Post-thaw motility and plasma membrane integrity of the frozen/thawed spermatozoa were 31.8 +/- 2.4% and 32.2 +/- 4.2%, respectively. The frozen/thawed spermatozoa were co-cultured with frozen/thawed immature homologous oocytes with intact ZP for 3 h to examine their ability to bind to the ZP. Sixteen of the 20 frozen/thawed sperm samples demonstrated the ability to bind to ZP. These results indicated that the freezing system for epididymal sperm used in the present study gives appropriate information for banking the genetic resources of wild felid species.     

Effects of mating on the termination of aggregation by the oak borer,Platypus quercivorus (Murayama) (Curculionidae: Platypodinae) during host colonization

We conducted a field experiment to determine whether logs bored by male Platypus quercivorus beetle were less attractive to conspecifics after beetles had mated. Bored (with male beetles) and unbored logs were placed in a beetle-infested forest and enclosed within a cage. For half of the bored logs, female beetles were released to mate with the male beetles. Log attractiveness was determined by assessing the number of beetles captured on adhesive paper placed on each cage enclosing the log. On the logs on which female beetles were released, the number of beetles captured decreased distinctly after release, and did not differ from the number captured on the unbored logs. In contrast, the number of beetles captured on logs with only unpaired males was higher than the number captured on the unbored logs. This result confirms that the attractiveness of logs bored by male P. quercivorus beetles declines after mating occurs. To our knowledge, this is the first evidence for the relaxation of aggregation pheromone following mating for beetles in the subfamily Platypodinae.