Heritability estimates of liabilities to digestive diseases in heavy racehorses

Data on 1410 heavy racehorses were taken from the Hokkaido Banei Horse Racing Result Book published during 1997–2000. Heritabilities were estimated using the restricted maximum likelihood method with a full data set of the accumulated prize winnings and the frequency of medical treatment against digestive diseases. Heritability estimates for the same traits were also obtained using a partial data set from affected animals only. In addition, a ‘two classes, one threshold’ model was applied to the full data set for the heritability estimation of liability to the diseases. Heritabilities estimated by restricted maximum likelihood with the full data set were quite low: 0.09 ± 0.02 for the prize winnings and 0.08 ± 0.03 for all three digestive diseases – colic resulting from flatulence, constipation and overfeeding. Low heritabilities were also estimated for each of the three kinds of colic. Medium heritabilities were estimated for colic resulting from constipation (0.30 ± 0.19) and overfeeding (0.46 ± 0.22) using the partial data set taken from the affected animals. Using the threshold model, higher heritabilities from 0.42 ± 0.04 to 0.57 ± 0.03 were estimated for the liabilities to all three digestive diseases and to the individual digestive diseases. These results suggest that liability evaluation of the frequency of medical treatment is insufficient because all non‐affected animals are scored as 0. The high heritabilities estimated by the threshold model with half‐sib relationships should be verified in future studies by comparing them with estimates from other genetic relationships and with realized heritabilities from a small‐scale practical selection program.     

Effects of protein phosphatase inhibitor calyculin a on the postacrosomal protein serine/threonine phosphorylation state and acrosome reaction in boar spermatozoa incubated with a cAMP analog

In order to reveal the involvement of the sperm postacrosomal region in the acrosome reaction, we examined the effects of the protein phosphatase inhibitor calyculin A on the postacrosomal protein serine/threonine phosphorylation state and acrosome morphology in boar spermatozoa incubated with a cAMP analog. Proteins were highly phosphorylated on the serine/threonine residues only in the postacrosomal region before incubation. After 90-min incubation without calyculin A, the protein phosphorylation state declined in the postacrosomal region irrespective of the capacitation state while it remained under the detectable level in the other regions of the sperm head. However, addition of calyculin A effectively suppressed the decline in protein phosphorylation state and increased an inactive form of protein phosphatase 1 in the postacrosomal region. On the other hand, this inhibitor had no influence on the protein phosphorylation state in the acrosome and equatorial segment. After incubation without calyculin A for 180 or 360 min, many spermatozoa exhibited acrosomal changes and loss that indicated occurrence of the acrosome reaction. However, addition of calyculin A significantly blocked these events. These results are consistent with our suggestion that postacrosomal serine/threonine-phosphorylated proteins are involved in suppression of the acrosome reaction in boar spermatozoa in vitro.     

Novel variations and loss of heterozygosity of BRCA2 identified in a dog with mammary tumors

OBJECTIVE: To establish novel polymorphic markers for analysis of loss of heterozygosity (LOH), so as to study the possible involvement of BRCA2 in mammary tumors obtained from dogs. SAMPLE POPULATION: Blood samples, mammary gland specimens, or mammary tumors from 3 tumor-bearing dogs and 10 tumor-free dogs. PROCEDURES: Nucleotide sequence analysis was performed with a DNA autosequencer. Loss of heterozygosity analysis was performed for markers established in the present study. The expression level of canine BRCA2 was quantified by real-time PCR analysis. RESULTS: 3 novel microsatellite markers with high heterozygosity rates (> 50%) were established, and the previously reported marker for canine BRCA2 gene locus was improved. These markers were used for the analysis of DNA from formalin-fixed and paraffin-embedded samples. By use of these markers, LOH in canine BRCA2 was identified as a result of recombination. In mammary tumor DNA that corresponded to the LOH-positive dog, the level of canine BRCA2 expression was decreased compared with that of nonneoplastic mammary gland tissue; the open reading frame contained 4 missense variations, 1 insertion variation, and 1 silent variation, some of which were localized to functional domains. CONCLUSIONS AND CLINICAL RELEVANCE: 3 novel polymorphic markers were developed for LOH analysis of canine BRCA2 and identified a dog with LOH with some variations in the functional domains. These markers could be useful for assessing the relevance of BRCA2 variation in mammary tumors of dogs.     

Surveillance of avian influenza viruses in Northern pintails (Anas acuta) in Tohoku District, Japan

Among winter migratory waterfowl, Northern pintails (Anas acuta), in one of the largest flocks in Tohoku district, northeast Japan, were surveyed for influenza A viruses at five wintering sites in three prefectures, viz., Aomori, Akita, and Miyagi. A total of 38 influenza A viruses were isolated from 2066 fecal samples collected during November 2006 through March 2007. The overall isolation rate was 1.84%. Eleven different subtypes were isolated, including nine H5N2, seven H6N8, seven H10N1, four H4N6, three H6N1, three H11N9, and one each of H1N1, H6N2, H6N5, H10N9, H11N1. Only the H4N6 subtype was detected during two successive months, November and December, from Lake Ogawara of Aomori prefecture. One wintering site, Lake Izunuma of Miyagi prefecture, was negative for virus isolation throughout the study period. During the sampling period, the highest virus isolation rate was in December (4.90%) followed by November (2.18%), January (0.91%), and February (0.30%). Virus isolation was negative for samples collected in March 2007. These results suggest that influenza viruses are introduced by Northern pintail when they migrate into Japan, but the viruses are not maintained in the flocks, most likely because the birds are not breeding during the winter. We believe that this relatively large data set creates a strong foundation for future studies of avian influenza virus (AIV) prevalence, evolution, and ecology in wintering sites, along with the role of Northern pintails in the spread of AIV during their migration from northern Russia and Asia to Japan.     

Comparisons of Ophiostomatoid fungi associated withTomicus piniperda andT. minor in Japanese red pine

Five species ofOphiostoma, twoLeptographium species and aGraphium species were isolated from two morphologically and ecologically similar bark beetle species,Tomicus piniperda andT. minor, and their infested Japanese red pine (Pinus densiflora) in Yamanashi Prefecture, central Honshu, Japan. An underscribedOphiostoma species andO. minus were isolated mainly fromT. piniperda and its galleries.Ophiostoma canum which was found for the first time in Japan was mainly fromT. minor and its galleries. Specific relationships between the beetles and fungal species are suggested. Contribution No.140, Laboratory of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba. Part of this study was presented at 108th Annual meeting of Japanese Forestry Society, April 2, 1997, Fukuoka, Japan.     

Small-scale variation in feeding environments for the Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> in a tidal flat in Tokyo Bay

The relative contribution of particulate organic matters (POMs) in water column and sediment as a food source for the Manila clam, Ruditapes philippinarum, was studied using carbon and nitrogen stable isotopic method (δ¹³C and δ¹⁵N) in a tidal flat at Seaside Park, Yokohama, Japan. Comparisons of δ¹³C and δ¹⁵N among R. philippinarum and POMs in surface water, bottom water, and sediment surface indicated that R. philippinarum larger than 5 mm shell length (SL) mainly assimilated benthic POM, and individuals smaller than 5 mm SL assimilated benthic and pelagic POM. Continuous measurements of chlorophyll concentrations in the bottom water revealed tide-driven resuspension of the benthic phytopigments. R. philippinarum showed differences in δ¹³C and δ¹⁵N along an inshore–offshore transect, indicating small-scale spatial differences in POM provision in the tidal flat. These findings suggest that POM in the bottom water, supposedly inhaled by R. philippinarum, is a mixture of a larger proportion of resuspended benthic POM and a smaller proportion of pelagic POM, and that the mixing ratio of the POMs may be affected by the hydrodynamics of flooding water associated with tidal flat topography.     

In search of the Kumamoto oyster <Emphasis Type="Italic">Crassostrea sikamea</Emphasis> (Amemiya, 1928) based on molecular markers: is the natural resource at stake?

A cornerstone in conserving wildlife is to resolve taxonomic uncertainties over organisms so that conservationists can define the entity that should be conserved. This is the case for two closely related Crassostrea oysters inhabiting the Ariake Sea (Kyushu, Japan) in sympatry, the kumamoto oyster C. sikamea and Pacific oyster C. gigas, where molecular markers have shed light on their taxonomic separation and species diagnosis. In the past, there was a concern that Kumamoto oysters may be at risk of extinction. However, several field surveys equipped with species-diagnosis molecular markers have provided solid evidence that this perception is incorrect. Nevertheless, previous studies have sometimes given conflicting results, throwing us into confusion about the resource status. Research on natural Kumamoto oysters that the author hopes could help to enhance our understanding of the conservation of this invaluable oyster is reviewed here.

Deformation and blemishing of pearls caused by bacteria

Bacteria cause deformation and blemishing in pearls, and we investigate this relationship. We examined pearls derived from Pinctada margaritifera, Pinctada maxima, and Pinctada fucata, and determined the location of bacteria using histopathological and immunohistochemical techniques and 16S ribosomal RNA (rRNA) sequence analyses. The most remarkable change was the inflammatory reaction located between the pearl nucleus and the nacreous layer, composed of hemocytic infiltration with melanization, periostracum, and fibrous aragonite-like structures. These anomalous changes were limited to abnormal sites, and such inflammatory reaction sites are a major factor in the formation of pearl abnormalities. Bacteria were detected from the inflammatory sites and are suspected as the causative agent. Most of these bacteria were anaerobic.     

Producibility and digestibility of antihypertensive beta-casein tripeptides, Val-Pro-Pro and Ile-Pro-Pro, in the gastrointestinal tract: analyses using an in vitro model of mammalian gastrointestinal digestion

Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) are antihypertensive tripeptides isolated from milk fermented with Lactobacillus helveticus and inhibit angiotensin-converting enzyme (ACE). We investigated whether these peptides were generated from beta-casein by digestive enzymes and whether they were resistant to enzymatic hydrolysis, using an in vitro model. VPP and IPP were not generated from beta-casein by gastrointestinal enzymes; instead, a number of longer peptides including VPP and IPP sequences were detected. The fermentation step would therefore be necessary to produce these antihypertensive tripeptides. VPP and IPP themselves were hardly digested by digestive enzymes, suggesting that orally administered VPP and IPP remain intact in the intestine, retaining their activity until adsorption. The present study also demonstrated that various functional peptide sequences in beta-casein were resistant to gastrointestinal enzymes. There may be a strong correlation between the resistance of peptides to gastrointestinal digestion and their real physiological effects after oral administration.