Species identification method for marine products of Seriola and related species

The complete nucleotide sequences of mitochondrial DNA (mtDNA) from four Seriola spp. (S. quinqueradiata, S. lalandi, S. dumerili, and S. rivoliana) were determined with the aim of developing a species identification analysis method for discriminating between commercially important Seriola spp. and other related species. In addition, the nucleotide sequences of the mitochondrial cytochrome b gene (Cytb) from five related but less expensive species in terms of market value (Seriolella brama, S. caerulea, S. punctata, Hyperoglyphe japonica, and Rachycentron canadum), which are often used as substitutes for Seriola spp., were determined. Restriction enzyme sites were examined by comparing the nucleotide sequences, and species-specific primers were designed for PCR-based restriction fragment length polymorphism (RFLP) analysis. Based on the results of the PCR amplification studies, the four Seriola spp. and the five related species tested could be categorized into three groups according to their PCR product pattern: a 373-bp product from the four Seriola spp., a 513-bp product from three Seriolella spp. and H. japonica, and a 204-bp product from R. canadum. In addition, RFLP analysis of the PCR products was able to differentiate these fish species.     

A comparison of the concentrations of C-reactive protein and alpha1-acid glycoprotein in the serum of young and adult dogs with acute inflammation

The concentrations of C-reactive protein (CRP) and alpha1-acid glycoprotein (AAG) were evaluated in 1-, 3- and 18-month-old dogs (four of each age) that had been inoculated with turpentine oil. The CRP and AAG in 3-month-old and younger dogs subjected to surgery or inoculated with either Staphylococcus aureus or a viral vaccine were also evaluated. The average CRP concentration in the sera peaked 2 days after inoculation of turpentine oil. The peak CRP concentrations in 3- and 18-month-old dogs were significantly (p < 0.05) greater than those in 1-month-old dogs. The average AAG concentration in the sera peaked 4 days after inoculation of turpentine oil. No significant difference was found in AAG concentrations between any of the age groups. When experimentally inoculated with S. aureus or subjected to oophorohysterectomy, the CRP and AAG concentrations increased in 3-month-old dogs, but they increased little in 1-month-old dogs. The CRP and AAG in dogs inoculated with the viral vaccine did not increase. In dogs with fractures or subjected to percutaneous gastrostomy, the CRP and AAG concentrations correlated with the condition of dogs.     

Rapid molecular typing of Listeria monocytogenes by pulsed-field gel electrophoresis

Pulsed-field gel electrophoresis (PFGE) is a highly discriminating tool for molecular typing, but the conventional PFGE protocol is time consuming. This paper describes a rapid method of PFGE for Listeria monocytogenes that yields results within 2 days.     

Virulence of ophiostomatoid fungi associated with the spruce bark beetleIps typographus f.japonicus in Yezo spruce

Yezo spruce trees (Picea jezoensis), approximately 40-year-old were inoculated with eight ophiostomatoid fungi associated withIps typographus f.japonicus to compare relative virulence of the fungi. Among them,Ophistoma penicillatum formed the longest necrotic lesion on inner bark around inoculation points, followed byO. aenigmaticum, Ceratocystis polonica, andO. bicolor, whileC. polonica formed a larger dry zone in sapwood than the other fungi. Yezo spruce trees were also mass inoculated withC. polonica, O. penicillatum, O. piceae singly or mixed to demonstrate the ability of the fungi to kill Yezo spruce trees. The trees inoculated withC. polonica, O. penicillatum or the mixed inoculum showed discoloration of needles in the early summer of the next year and died by autumn. However, the trees inoculated withO. piceae or the control inocula did not die, except for one tree. These results indicated thatC. polonica andO. penicillatum were more virulent thanO. piceae and suggested that they might be at least partially responsible for the mortality of the beetle-infested Yezo spruce trees. Part of this study was supported by the Sumitomo Foundation, Japan to Y. Yamaoka and I. Takahashi. Part of this study was presented at the 107th meeting of the Japanese Forestry Society, April 1–4, 1996, Tsukuba, Ibaraki, at the 42nd annual meeting of the Mycological Society of Japan, May 16–17, 1998, Kyoto, and at the 110th meeting of the Japanese Forestry Society, April 2–5, 1999, Matsuyama, Ehime. Contribution No. 143, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Morphological analysis of a deep-sea whelk <Emphasis Type="Italic">Buccinum tsubai</Emphasis> in the Sea of Japan

ABSTRACT:   Morphometrical analyses were performed on Buccinum tsubai , a deep-sea gastropod found in the Sea of Japan, and morphological differences were examined between the gender and among the four local subpopulations of Hokkaido, Yamagata-Toyama, the Yamato Bank, and San'in which are distinguished by mitochondrial DNA sequence analysis. As a result, sexual dimorphism of B. tsubai was found. Morphological differences were also recognized among four areas which may be related to genetic differences. They are also thought to be associated with the phenotypic plasticity in response to different environmental factors in each area. B. tsubai inhabits the the upper portion of the Sea of Japan Proper Water (UJSPW); and in the UJSPW of the Yamato Basin, nutrients are comparatively abundant and the dissolved oxygen content is low. Such environmental differences may be related to morphological differences among the four areas.     

Comparison of estrogen responsive genes in the mouse uterus, vagina and mammary gland

Female reproductive organs are mainly regulated by estrogen and progesterone. Specifically, the uterus, vagina and mammary gland show organ-specific mitosis and morphological changes during proliferative events, such as estrous cycle, gestation and lactation. The mechanism underlying these organ-specific estrogen-dependent events is still unknown. We examined, therefore, global gene expression in the mature uterus, vagina and mammary gland of ovariectomized adult mice 6 hr after an injection of 5 microg/kg 17beta-estradiol (E2) using a microarray method in order to identify primary E2-responsive genes. Half of the E2 up-regulated genes in the uterus were similar to those in the vagina. E2 up-regulated the expression of Insulin-like growth factor 1 (Igf-1) genes in the uterus and vagina. In the vagina, E2 up-regulated the expression of IGF binding proteins (Igfbp2 and Igfbp5). In the mammary gland, unlike the uterus and vagina, no gene showed altered expression 6 hr after the E2 exposure. These results suggest that expression of Igf-1 and morphogenesis genes is regulated by E2 in an organ-specific manner, and it is supported by the results of BrdU labeling showing E2-induced mitosis in the uterus and vagina except the mammary gland. The differences in organ specificity in response to E2 may be attributed by differences in gene expression regulated by E2 in female reproductive organs. The candidate estrogen-responsive genes in the uterus and vagina identified by profiling provide an important foundation understanding functional mechanisms of estrogen regulating morphogenesis and maintenance of each reproductive organ.     

Utility of 5-aminolaevulinic acid fluorescence-guided endoscopic biopsy for malignant mesothelioma in a cat and dog

Malignant mesothelioma (MM) is uncommon in cats and dogs and can be challenging to diagnose. Adequate tissue sampling is required for superior diagnostic accuracy. Protoporphyrin IX, a metabolite of 5-aminolaevulinic acid (5-ALA), is a photosensitiser for photodynamic diagnosis (PDD). To the best of our knowledge, no study has reported the use of 5-ALA-PDD to detect MM in veterinary medicine. The present study describes the use of 5-ALA-PDD for MM diagnosis in a cat and dog, as well as the effectiveness of intracavitary chemotherapy. We evaluated the use of PDD with 5-ALA hydrochloride (5-ALA-PDD) in two cases of MM. A 12-year-old cat presented with a 1-month history of respiratory distress, and a 9-year-old dog presented with a 3-month history of mild abdominal distention. We endoscopically biopsied lesions in both the cases using 5-ALA-PDD. Histopathological examination revealed mesothelioma, and immunohistochemical staining was positive for calretinin. Both patients were treated with carboplatin. The cat died of respiratory failure. Although, the dog's condition improved 21 days after the first chemotherapeutic drug administration, the dog died on day 684 owing to cardiac-related issues. 5-ALA-PDD is thus, safe and feasible for the diagnosis of MM in veterinary medicine.     

Effects of reaggregated granulosa cells and oocytes derived from early antral follicles on the properties of oocytes grown in vitro

In this study, we examined the effects of reconstructed oocyte–granulosa cell complexes (OGCs) on the development of porcine oocytes derived from early antral follicles (EAFs; 0.5–0.7 mm in diameter). When denuded oocytes were cocultured with granulosa cells derived from other EAFs, the oocytes and granulosa cells aggregated to form OGCs after 2 days of culture. After 14 days of culture, we compared cell number, oocyte diameter, and oocyte chromatin configuration in unmanipulated (natural) OGCs, reconstructed OGCs, and OGCs collected from antral follicles (AFs, 3.0–6.0 mm in diameter). The diameters of oocytes from reconstructed OGCs grown in vitro were not different from those of oocytes from natural OGCs, although they were significantly smaller than those of oocytes from antral follicle (AF) OGCs. Oocyte chromatin configuration did not differ among the 3 OGC groups, but the oocyte nuclear maturation rate was lower in the reconstructed OGCs and higher in the AF OGCs. However, when the in vitro culture period for the reconstructed OGCs was extended by 2 days, the nuclear maturation rate of oocytes from reconstructed OGCs was similar to that of oocytes from natural OGCs. In addition, blastocysts were successfully obtained from oocytes from reconstructed OGCs. In conclusion, we established an innovative culture method that allows oocytes and granulosa cells from EAFs to reaggregate as reconstructed OGCs, which yield oocytes with the ability to develop to the blastocyst stage.