Phylogenetic analysis of noxious red tide flagellates Chattonella antiqua, C. marina, C. ovata, and C. verruculosa (Raphidophyceae) based on the rRNA gene family

ABSTRACT:   Four species of Chattonella , which are well known to form red tides that are lethal to fish, were subjected to phylogenetic analysis on the basis of the ribosomal RNA genes (rDNA), 5.8S rDNA, 18S rDNA, 28S rDNA, and the flanking internal transcribed spacers 1 and 2 (ITS1 and ITS2). The 18S rDNA sequences of C. antiqua , C. marina , and C. ovata isolated from different regions in Japan were compared. They were found to be identical with each other in a sequence 1818 bp long. The sequences of the D1/D2 region in the 28S rDNA, 5.8S rDNA, and ITS region that are known to be more variable regions were also found to be identical. These homogeneities of the rRNA gene family revealed the extremely close relatedness of C. antiqua , C. marina , and C. ovata . The sequences of C. verruculosa were different from those of these three species , resulting in an 89.2% homology in the 18S rDNA sequences, 70.4% homology in the D1/D2 region in the 28S rDNA sequences, and an 81.5% homology in 5.8S rDNA sequences and the ITS regions. Chattonella verruculosa was grouped within a single cluster composed of Dictyochophyceae rather than the other species of Raphidophyceae.     

Effect of danazol on NK cells and cytokines in the mouse uterus

Danazol, which has been used as a medicine for endometriosis, has a valid effect in pretreatment of patients receiving in vitro fertilization and embryo transfer, although its reproductive mechanism remains unclear. BALB/c mice were subcutaneously injected with danazol for 2 weeks. Blood and uteri were collected and cytokines were assayed. Following danazol treatment, an increase in pregnancy ratio was evident that was accompanied by up-regulation in serum macrophage-colony stimulating factor (M-CSF). RT-PCR analysis revealed that expression of M-CSF and Ly49, a phenotypic marker of natural killer (NK) cells, was up-regulated in the uteri of the danazol-treated mice. In immunohistochemical analysis, M-CSF and Ly49, together with alpha5 integrin, were clearly detected in the endometrium of the danazol-treated mice with very similar pattern of localization. These results suggest that danazol has an effect to promote pregnancy that induces recruitment of NK cells and a concomitant increase in the expression of M-CSF and alpha5 integrin in the uterus.     

The formation process of button ulcers in pigs experimentally infected with a subgenotype 2.1 isolate of classical swine fever virus

We evaluated the role of classical swine fever virus (CSFV) in the formation of button ulcers in the mucosa of the gastrointestinal tract. Histopathological and immunohistochemical analyses of pigs experimentally infected with a subgenotype 2.1 isolate of CSFV, which was isolated in Japan in 2019, revealed follicular necrosis in the submucosal mucosa-associated lymphoid tissue and herniation of crypts as factors that contribute to the development of button ulcers during CSFV infection. These findings indicate that CSFV induces follicular necrosis and is one of the causative agents of button ulcers in pigs.     

Effects of dietary heme iron and exercise training on abdominal fat accumulation and lipid metabolism in high‐fat diet‐fed mice

Animal by‐products can be recycled and used as sources of essential nutrients. Water‐soluble heme iron (WSHI), a functional food additive for supplementing iron, is produced by processing animal blood. In this study, we investigated the effects of dietary supplementation of 3% WSHI and exercise training for 4 weeks on the accumulation of abdominal fat and lipid metabolism in mice fed high‐fat diet. Exercise‐trained mice had significantly less perirenal adipose tissue, whereas WSHI‐fed mice tended to have less epididymal adipose tissue. In addition, total weight of abdominal adipose tissues was significantly decreased in the Exercise + WSHI group. Dietary WSHI significantly increased the messenger RNA (mRNA) levels of lipoprotein lipase and hormone‐sensitive lipase. WSHI‐fed mice also tended to show increased mRNA levels of adipose triglyceride lipase in their epididymal adipose tissue. Dietary WSHI also significantly decreased the mRNA levels of fatty acid oxidation‐related enzymes in the liver, but did not influence levels in the Gastrocnemius muscle. Exercise training did not influence the mRNA levels of lipid metabolism‐related enzymes in the epididymal adipose tissue, liver or the Gastrocnemius muscle. These findings suggest that the accumulation of abdominal fat can be efficiently decreased by the combination of dietary WSHI and exercise training in mice fed high‐fat diet.     

Evaluation of the juice brix of wild sugarcanes (Saccharum spontaneum) indigenous to Japan

Modern sugarcane cultivars are derived from the interspecific crossing between Saccharum officinarum and wild sugarcane, Saccharum spontaneum. The introgression of valuable characteristics from wild sugarcane is recognized as extremely important, but this process typically requires long-term effort over multiple generations of backcrosses owing to the low sugar content of the initial interspecific hybrids. In this study, we aimed to identify Japanese wild sugarcanes with high juice brix in order to promote effective interspecific crossing of sugarcane. Sixty-four accessions from the Nansei Islands and 70 accessions from the Honshu were evaluated for juice brix. Wild sugarcanes with high juice brix were demonstrated to exist among wild sugarcanes indigenous to the Honshu. A significant difference was observed between the median juice brix values of wild sugarcanes of the Nansei Islands and those of the Honshu. The relationship between juice brix and stem traits was then examined in 20 wild sugarcanes, 10 each from the Nansei Islands and the Honshu. The reproducibility of juice brix value in both experiments was confirmed. In contrast to juice brix, stem traits, such as length, diameter, and volume, were typically smaller in wild sugarcanes from the Honshu. Moreover, a negative correlation was observed between the index of stem volume and juice brix. In this study, we identified outstanding wild sugarcanes with high juice brix. Using germplasms from the identified wild sugarcanes in interspecific crossing could contribute to the increases in both yield and sugar content.     

Fish polyculture system integrated with vegetable farming improves yield and economic benefits of small‐scale farmers

This study explored the yield and economic benefits of African sharptooth catfish, Clarias gariepinus, and Nile tilapia, Oreochromis niloticus polyculture, reared in earthen ponds for 270 days integrated with Chinese cabbage, Brassica rapa chinensis, farmed for 45 days to small‐scale farmers in Tanzania. An integrated aquaculture–agriculture (IAA) system involving fishes and vegetables resulted in three and 2.5 times higher net yield than the culture of fishes alone and farming of non‐integrated vegetables respectively. Vegetables irrigated with pond water resulted in 1.8 times higher net and annual yields than those irrigated with stream water. The practice of fish–vegetable integration and growing of vegetables irrigated with pond water produced 14 and 13 times higher net annual yield, respectively, than the culture of fishes alone. The integrated system produced 20 and 150 times more net returns compared with vegetables irrigated with pond and stream water respectively. Integrating fishes with vegetables resulted in significantly higher annual net cash flow than the farming of fishes and vegetables separately. The study revealed that integrating fishes with B. rapa chinensis increases yields, income and food production to small‐scale farmers. Thus, small‐scale farmers can adopt IAA technology to meet the increasing demand of fish and vegetables, stabilize their incomes and diversify food production, consequently improving food security. These benefits will be realized by providing proper training in IAA technology and management skills to small‐scale farmers.     

PCR-DGGE analysis of bacterial communities in <Emphasis Type="Italic">funazushi</Emphasis>, fermented crucian carp with rice,during fermentation

Funazushi (fermented Crucian Carp with rice) is a fermented fish product found only around Lake Biwa in Shiga Prefecture, Japan. It is characterized by a unique cheese-like flavor and characteristic sour taste. We analyzed the changes in the microbial community during funazushi fermentation by denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA fragments (PCR-DGGE) and by plate counts. The plate counts showed that lactic acid bacteria reached 8.0 log₁₀ CFU/g within 7 days of fermentation initiation before decreasing slowly to 4.0 log₁₀ CFU/g during the remainder of 1-year study period. PCR-DGGE revealed that the dominant bacteria in the initial (days 14 and 30) and latter (days 90, 180, and 360) periods of fermentation were Lactobacillus plantarum and L. acetotolerans. This is the first identification of L. acetotolerans in funazushi as traditional cultivation techniques have not been sufficiently sensitive. This is the first report of PCR-DGGE being used to assess the microbial community in funazushi. This technique was also found to be effective in profiling microbial diversity.