The transition time from the juvenile phase to the adult phase differs in soybean cultivars ‘Enrei’ and ‘Peking’

Plants develop juvenile phase to adult phase in vegetative stage. Although soybean is a very important crop worldwide, there has been only one study of the juvenile–adult phase change. In this study, we determined that the juvenile–adult phase change occurred at different stages in two soybean cultivars that differ in their photosensitivity. Cultivar ‘Enrei’ (E1e2e3E4) is weakly photosensitive and cultivar ‘Peking’ (E1E2E3E4) is strongly photosensitive. In ‘Enrei’, the leaf size gradually increased at a constant leaf position regardless of the difference in day length. In ‘Peking’ plants transferred to short‐day conditions at several leaf development stages, leaf size gradually increased at different leaf positions. Expression of miR156 by ‘Enrei’ transferred to short‐day conditions had nearly the same pattern as that of ‘Enrei’ grown under long‐day conditions. In ‘Peking’, the expression of miR156 had different patterns in younger leaves of plants subjected to either a short‐day treatment or long‐day conditions. These results indicate that the E2 and E3 loci that regulate photosensitivity also regulate the expression of miR156 and the juvenile–adult phase change in soybean.     

Phylogenetic groups and cephalosporin resistance genes of Escherichia coli from diseased food-producing animals in Japan

A total of 318 Escherichia coli isolates obtained from different food-producing animals affected with colibacillosis between 2001 and 2006 were subjected to phylogenetic analysis: 72 bovine isolates, 89 poultry isolates and 157 porcine isolates. Overall, the phylogenetic group A was predominant in isolates from cattle (36/72, 50%) and pigs (101/157, 64.3%) whereas groups A (44/89, 49.4%) and D (40/89, 44.9%) were predominant in isolates from poultry. In addition, group B2 was not found among diseased food-producing animals except for a poultry isolate. Thus, the phylogenetic group distribution of E. coli from diseased animals was different by animal species. Among the 318 isolates, cefazolin resistance (minimum inhibitory concentrations: ≥32 μg/ml) was found in six bovine isolates, 29 poultry isolates and three porcine isolates. Of them, 11 isolates (nine from poultry and two from cattle) produced extended spectrum β-lactamase (ESBL). The two bovine isolates produced bla_CTX-M-2, while the nine poultry isolates produced bla_CTX-M-25 (4), bla_SHV-2 (3), bla_CTX-M-15 (1) and bla_CTX-M-2 (1). Thus, our results showed that several types of ESBL were identified and three types of β-lactamase (SHV-2, CTX-M-25 and CTX-M-15) were observed for the first time in E. coli from diseased animals in Japan.     

Annual Element Budget of Soil in Snow-Dominated Forested Ecosystem

Seasonal fluctuation of concentration and flux of major inorganic ions in throughfall, stem flow, snowpack and soil solution was investigated at a natural cool temperature mixed forest in Hokkaido, northern Japan, in order to clarify the effect of snowmelt on the solute dynamics in the forest soil in snow-dominated region. Na⁺, Ca²⁺, Mg²⁺, Cl^? and SO₄ ^2? concentrations in soil solution showed a large fluctuation in the snowmelt period. The percentage of output of these elements from soil during the snowmelt period in the annual output was as follows. Mg²⁺: 51%, Na⁺ and Cl^?: 59 and 60%, SO₄ ^2?: 65%, Ca²⁺: 77%. Our results indicated that the snowmelt event was very important to quantify the annual elemental budgets in this region. Although the leaching of base cation from the soil was larger than that of inputs and accumulation into the vegetation, annual decreasing rate of acid neutralization capacity (ANC_(s)) from the soil was mostly affected by the base cation accumulation into the vegetation, related that the base cations weathering accompanied with bicarbonate was slow due to the acidic and weathered soil in the studied site. It is suggested that the weakly acidic soil which has low ANC_(s) in snow-dominated region will be relatively sensitive to the future increase of acidic deposition.     

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.     

Nuclear Replacement of In Vitro‐Matured Porcine Oocytes by a Serial Centrifugation and Fusion Method

The objective of the present study was to establish a method for nuclear replacement in metaphase‐II (M‐II) stage porcine oocytes. Karyoplasts containing M‐II chromosomes (K) and cytoplasts without chromosomes (C) were produced from in vitro‐matured oocytes by a serial centrifugation method. The oocytes were then reconstructed by fusion of one karyoplast with 1, 2, 3 or 4 cytoplasts (K + 1C, K + 2C, K + 3C and K + 4C, respectively). Reconstructed oocytes, karyoplasts without fusion of any cytoplast (K) and zona‐free M‐II oocytes (control) were used for experiments. The rates of female pronucleus formation after parthenogenetic activation in all groups of reconstructed oocytes (58.2–77.4%) were not different from those of the K and control groups (58.2% and 66.0%, respectively). In vitro fertilization was carried out to assay the fertilization ability and subsequent embryonic development of the reconstructed oocytes. The cytoplast : karyoplast ratio did not affect the fertilization status (penetration and male pronuclear formation rates) of the oocytes. A significantly high monospermy rate was found in K oocytes (p < 0.05, 61.6%) compared with the other groups (18.2–32.8%). Blastocyst formation rates increased significantly as the number of the cytoplasts fused with karyoplasts increased (p < 0.05, 0.0–15.3%). The blastocyst rate in the K + 4C group (15.3%) was comparable with that of the control (17.8%). Total cell numbers in both the K + 3C and K + 4C groups (16.0 and 15.3 cells, respectively) were comparable with that of the control (26.2 cells). Our results demonstrate that a serial centrifugation and fusion (Centri‐Fusion) is an effective method for producing M‐II chromosome transferred oocytes with normal fertilization ability and in vitro development. It is suggested that the number of cytoplasts fused with a karyoplast plays a critical role in embryonic development.     

Detection and quantification of protein residues in food grade amino acids and nucleic acids using a dot-blot fluorescent staining method

Food allergies represent an important health problem in industrialized countries, such that detection and quantitative analysis of the protein considered to be the main allergen is crucial. A dot-blot fluorescent staining method for the detection and quantitative analysis of protein residues in food grade amino acids and nucleic acids is presented. This method combines fluorescence staining with dot-blotting onto PVDF membrane. Several standard proteins, such as bovine serum albumin (66 kDa), lysozyme (14 kDa), ubiquitin (8.6 kDa), bovine insulin (5.7 kDa), and oxidized insulin B chain (3.5 kDa), were detectable at 0.1 ppm using SYPRO Ruby blot stain. Twenty-five different amino acids and two different nucleic acids of food grade were analyzed using this method combined with an internal standard addition method using BSA as an internal standard. All amino acids and nucleic acids were dissolved in 3.6% aqueous HCl and dot-blotted onto PVDF membrane before a large amount of amino acids and nucleic acid were removed. Protein residues and the internal standard protein immobilized on the membrane were stained using SYPRO ruby blot stain. The internal standard in all samples was detectable at 0.1 ppm. Samples were dissolved at 120 or 70 mg/mL, according to their solubility under acidic conditions. The detection limit of protein residues per weight was 0.8-1.4 ppm in amino acids and nucleic acids; residual protein was not detected in any sample.     

Increase in the Nitrogen Content of Soil by the Introduction of Earthworms into Soil

The contribution of an earthworm species ( Amynthas vittatus ) to the increase of the nitrogen content of soil was examined. Three specimens of adult earthworms were introduced into 300 g of soil (Gray Lowland soil, silty clay) supplemented with 1% carboxymethyl cellulose in a container and incubated for 32 d at 22°C in the dark. The contents of total-N, NH₄-N and NO₃-N, and the population of aerobic nitrogen-fixing bacteria in soil significantly increased after incubation with the earthworms, while the natural abundance of ¹⁵N (δ¹⁵N) in soil decreased. The amount of nitrogen in the earthworms did not decrease during the incubation in the microcosm. Both acetylene reduction activity of the microcosm and incorporation of ¹⁵N to soil from atmospheric ¹⁵N₂ were significantly enhanced by the introduction of the earthworms into soil, though the observed increment of nitrogen in soil was much higher than the estimated one based on the nitrogen-fixing activity. The results obtained in the present study indicated that the earthworms increased the nitrogen content of soil, presumably due to the enhancement of the nitrogen-fixing activity of the soil from the microcosm by the earthworms.     

Goat milk acceptance and promotion methods in Japan: The questionnaire survey to middle class households

A consumer questionnaire conducted with the purpose of ascertaining the acceptability of goat milk and related products in Japan was carried out on 345 guarantees of Nippon Veterinary and Life Science University in December 2006. 275 effective responses (79%) representing middle class urban households were returned. The results revealed that (1) 30% of respondents have experienced drinking goat milk and only 10% are aware of the current retail situation of goat milk and related products; (2) over 70% of goat milk drinkers raised goats by hand at some point in their past and their first experience drinking goat milk was in infancy; (3) those with experience in drinking goat milk expressed a vague evaluation and minimal understanding of drinking goat milk; (4) respondents who were inexperienced goat milk drinkers expressed a strong desire to taste and a weak desire to purchase goat milk; (5) respondents expressed low recognition regarding retailed goat milk products, but those who had already purchased goat milk products expressed a high evaluation and strong desire to purchase these products again; and (6) recognition of goat milk characteristics is low, but those with high recognition also rate goat milk highly. Goats are perceived as being 'mild and familiar.' It is necessary for those who manage goat husbandry to present goat milk and related product tasting opportunities to consumers. The key point is to make the functional differences between cow and goat milk clear and present the advantages of goat milk at the fore of this promotion. Goat milk should not be promoted merely as a drink that is similar to cow milk, but must be positioned as a functional drink or health food in order to expand the Japanese goat milk market.     

Nucleotide sequence homology to bovine viral diarrhea virus 2 (BVDV 2) in the 5′ untranslated region of BVDVs from cattle with mucosal disease or persistent infection in Japan

Cytopathogenic and non-cytopathogenic bovine viral diarrhea viruses (BVDVs) were isolated from cattle with mucosal disease or persistent infection in Japan. These isolates were compared for antigenic properties by cross-neutralization tests with Japanese reference strains of BVDV belonging to classical type 1. Significantly low cross-reactivity to reference strains was noted, indicating the viruses to possibly represent a new serotype in Japan. Thus, to determine the genotype of the isolates, nucleotide sequences of the 5′ untranslated region were determined and compared with those of previously reported BVDV 1 and 2. The isolates were clearly shown to belong to BVDV 2, not to BVDV 1.