Comparison of the amount of thiotrophic symbionts in the deep-sea mussel <Emphasis Type="Italic">Bathymodiolus septemdierum</Emphasis> under different sulfide levels using fluorescent in situ hybridization

Various invertebrates inhabiting hydrothermal vents harbor thiotrophic endosymbionts that provide the host with nutrients and are possibly involved in the detoxification of harmful sulfides. In this study, we first determined the partial 16S rRNA gene sequence of the thiotrophic symbiont of the deep-sea mussel Bathymodiolus septemdierum, a dominant species at hydrothermal vents in the Izu–Ogasawara (Bonin) area. We then designed a new probe, Bsob692, for fluorescent in situ hybridization (FISH) using regions completely conserved among thiotrophic symbionts of all bathymodiolin mussels and established the protocol for FISH to compare the distribution and amount of the symbiont using an image analysis program that is commercially available. We compared fluorescent intensity in the gill of the mussels collected at different sites and found a higher intensity in specimens collected from a site with higher sulfide concentration. We also compared mussels reared in the presence and absence of sulfide and found that the former had a higher fluorescent intensity.     

‘Niigata S3’ is a new strawberry cultivar suitable for forcing culture under low temperature and insolation conditions

‘Niigata S3’ is a new strawberry (Fragaria × ananassa Duch.) cultivar that is early flowering and possesses high soluble solid content and good coloration. It was selected from a cross between Kei812 (seed parent) and ‘Asuka-Ruby’ (pollen parent). The first harvest date of ‘Niigata S3’ was December 27, 34 days earlier than ‘Echigohime’ and 9 days earlier than ‘Asuka-Ruby’ (means of 2007 and 2008). The marketable yield of ‘Niigata S3’ was 85% of ‘Echigohime’, 107% of ‘Asuka-Ruby’, while the early yield was 145% of ‘Echigohime’, 85% of ‘Asuka-Ruby’ (based on 2007 and 2008 means). The shape of the fruit is long conical, and its skin color medium-red. The fruit skin hardness of ‘Niigata S3’ was 31.5 g/mm², which was harder than ‘Echigohime’, and its average soluble solid content was 11.4%, which was higher than the values for ‘Echigohime’ and ‘Asuka-Ruby’ (2008). Furthermore, ‘Niigata S3’ did not bear apical overripe fruit. This new cultivar is adaptable to the climatic conditions of Niigata, as well as other regions that experience low winter temperatures and insolation.     

Proper reprogramming of imprinted and non‐imprinted genes in cloned cattle gametogenesis

Epigenetic abnormalities in cloned animals are caused by incomplete reprogramming of the donor nucleus during the nuclear transfer step (first reprogramming). However, during the second reprogramming step that occurs only in the germline cells, epigenetic errors not corrected during the first step are repaired. Consequently, epigenetic abnormalities in the somatic cells of cloned animals should be erased in their spermatozoa or oocytes. This is supported by the fact that offspring from cloned animals do not exhibit defects at birth or during postnatal development. To test this hypothesis in cloned cattle, we compared the DNA methylation level of two imprinted genes (H19 and PEG3) and three non‐imprinted genes (XIST, OCT4 and NANOG) and two repetitive elements (Satellite I and Satellite II) in blood and sperm DNAs from cloned and non‐cloned bulls. We found no differences between cloned and non‐cloned bulls. We also analyzed the DNA methylation levels of four repetitive elements (Satellite I, Satellite II, Alpha‐satellite and Art2) in oocytes recovered from cloned and non‐cloned cows. Again, no significant differences were observed between clones and non‐clones. These results suggested that imprinted and non‐imprinted genes and repetitive elements were properly reprogramed during gametogenesis in cloned cattle; therefore, they contributed to the soundness of cloned cattle offspring.     

Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan

Compared with other countries, surveys of these parasites have been rarely performed in companion animals of Japan in spite of their significance for public health. Here, we investigated pet dogs and cats in Japan for the first time, and genetically analyzed the isolates to evaluate the risk of zoonotic infections. Seventy-seven fecal samples were collected from privately owned dogs and 55 samples from owned cats in Osaka city, Japan. Cryptosporidium oocysts were identified in 3/77 dogs (3.9%) and 7/55 cats (12.7%), and Giardia infection in 2/77 dogs (2.6%) and 1/55 cats (1.8%). Amplification of the target regions for genotyping was successful, Cryptosporidium isolates in dogs and cats were identified as C. canis and C. felis, respectively, and those of Giardia in dogs and cats were G. intestinalis Assemblages D and F. The discharge period of the oocysts varied within 3-16 weeks and that of the cysts was 12 weeks. To date, zoonotic types of both parasites have been identified in other animals in Japan, and further large-scale studies are needed to determine the distribution of zoonotic genotypes in these animals, especially those closely associated with humans.     

A comparative analysis of acceptance by Japanese females and price of goat milk from different sources

A sensory survey was carried out using 4 different types of whole goat milk among middle‐class females to investigate consumer acceptability of goat milk and whether there is an opportunity to expand the sale of goat milk products in Japan. Four different types of whole milk powder (domestic concentrate‐fed, domestic pasture‐fed, USA commercially canned, and New Zealand commercially canned) were used. Fresh cow milk was served as a control. Thirty‐one housewives evaluated the 5 liquid milk samples for smell, taste and overall characteristics on a scale of 1 (low) to 3 (high). Chi‐square analyses were carried out to detect significant differences between the milk types in each category. The goat milk from the USA was the most preferred goat milk with respect to smell and overall evaluation. Domestic pasture‐fed milk received the lowest grade in the evaluation for its ‘grassy and goaty’ smell. This result shows us that pasture intake affects the taste and smell of powdered milk which gives the lowest evaluation by the participants. If Japanese goat milk producers want to be successful in the domestic goat milk market and compete against goat milk products from other countries, they should improve production methods and flavor of their products.     

Metabolic discrimination of mango juice from various cultivars by band-selective NMR spectroscopy

NMR-based metabolic analysis of foods has been widely applied in food science. In this study, we performed discrimination of five different mango cultivars, Awin, Carabao, Keitt, Kent, and Nam Dok Mai, using metabolic analysis with band-selective excitation NMR spectra. A combination of unsupervised principal component analysis (PCA) with low-field region (1)H NMR spectra obtained by band-selective excitation provided a good discriminant model of the five mango cultivars. Using F(2)-selective 2D NMR spectra, we also identified various minor components in the mango juice. Signal assignment of the minor components facilitated the interpretation of the loading plot, and it was found that arginine, histidine, phenylalanine, glutamine, shikimic acid, and trigonelline were important for classification of the five mango cultivars.     

Nutritional characteristics of the leaves of native plants growing in adverse soils of humid tropical lowlands

Acid sulfate, peat, sandy podzolic, and saline soils are widely distributed inthe lowlands of Thailand and Malaysia. The nutrient concentrations in theleaves of plants grown in these type of soils were studied with the aim ofdeveloping a nutritional strategy for adapting to such problem soils. In sagoand oil palms that were well-adapted to peat soil, the N, P, and Kconcentrations were the same in the mature leaves, while the Ca, Mg, Na,and Fe concentrations were higher in the mature leaves of the oil palm thanof the sago palm. Melastoma malabathricum and Melaleuca cajuputi plantsthat were well-adapted to low pH soils, peat, and acid sulfate soils were alsostudied. It was observed that a high amount of Al accumulated in the M.marabathricum leaves, while Al did not accumulate in M. cajuputi leaves.M. cajuputi plants accumulated large amounts of Na in their leaves or stemsregardless of the exchangeable Na concentration in the soil, while M.malabathricum that was growing in saline-affected soils excluded Na.Positive relationships between macronutrients were recognized between Pand N, between K and N, and between P and K. Al showed antagonisticrelationships with P, K, Ca, Mg, Fe, Zn, Cu, and Na. Na also showedantagonistic relationships with P, K, Zn, Mn, Cu, and Al. Fe showed weakantagonistic relationships with Zn, Mn, Cu, and Al.     

Neutropenia associated with osteomyelitis due to Hepatozoon canis infection in a dog

A 4-year-old, intact male Shiba dog was referred to Yamaguchi University Animal Medical Center, Yamaguchi, Japan, for the following complaints: anorexia, lethargy, intermittent fever, gingival bleeding and abdominal purpura. The dog presented with persistent neutropenia. Histopathological examination of a bone marrow sample revealed round to oval structures that resembled Hepatozoon micromerozoites and formed a "wheel-spoke" pattern. Furthermore, mature neutrophils were observed around these structures. PCR and sequencing using bone marrow aspirate confirmed Hepatozoon canis (H. canis) infection. These findings suggest that the neutropenia observed in this case was associated with osteomyelitis due to H. canis infection. This is the first report of neutropenia associated with H. canis infection. H. canis infection can be included in the differential diagnosis in canine cases of neutropenia in areas where the disease is endemic.     

Molecular and phenotypic characteristics of CMY-2 β-lactamase-producing Salmonella enterica serovar Typhimurium isolated from cattle in Japan

Isolates of extended-spectrum cephalosporin (ESC)-resistant Salmonella enterica serovar Typhimurium obtained from two different farms in Fukushima Prefecture, Japan, in 2007 were characterized in order to determine the genetic basis of resistance. ESC resistance in the two isolates was mediated by an AmpC β-lactamase encoded by the bla(CMY-2) gene, which is located in a large self-transmissible plasmid in each isolate. The sizes of the bla(CMY-2)-carrying plasmids were different. The replicon types of the plasmids were I1-Iγ and A/C. The results of macrorestriction analysis and phage typing suggest a close relationship between both isolates. This is the first report of ESC-resistant S. Typhimurium isolated from cattle in Japan.