Ontogeny of immune system organs in northern bluefin tuna (Thunnus orientalis, Temminck and Schlegel 1844)

Serial sections, prepared from 0.5 to 30 days posthatch (dph) larval and juvenile Thunnus orientalis (Temminck & Schlegel 1844), were stained with haematoxylin and eosin and examined by light microscopy for immune organ development. The early kidney was present at 0.5 dph, undifferentiated stem cells began to appear at 2 dph, and by 7 dph occasional small lymphocytes were present. The thymus was first obvious at 5 dph, located above the fourth branchial arch, small lymphocytes appeared at 7 dph, and by 15 dph an outer thymocytic zone and an inner epithelioid zone were visible. The progenitor spleen was present at 2 dph, located close to the gut, and by 12 dph it consisted of a mass of sinusoids filled with red blood cells, and remained mainly erythroid throughout the period studied. These results suggest that development of immune organs in this species is precocious relative to other marine teleosts.     

Rhizobium Proliferation-Supporting Substances in Seed Exudates of Common Bean (Phaseolus vulgaris L.)

Seed exudates collected from three cultivars (cv. Hokkaikintoki, Himetebou, and Kurodane-kinugasa) of common bean ( Phaseolus vulgaris L.) were fractionated based on several physicochemical properties, and the nutrient substances in the fractions effective for Rhizobium proliferation were determined. Regardless of the common bean cultivars, the low molecular weight and high polarity fraction showed the most beneficial effect on Rhizobium proliferation. After further separation of the low molecular weight and high polarity fraction, the promoting effect was detected in the non-ionic and the cationic plus ampho-teric subtractions. Although the amount of sugars or amino acids exuded from a seed was, respectively, similar in all the cultivars except for the sugars in Kurodanekinugasa, the content of these compounds per seed-weight was inversely proportional to the seed size. Ten sugars and twenty-one amino acids were, respectively, identified in the non-ionic and cat-ionic plus amphoteric subtractions, and there were some differences in the composition of these substances between common bean cultivars.     

Single-copy nuclear PolA1 gene sheds light on the origin of S genome with relationships to B and G genomes of polyploid wheat species

PolA1, a single-copy nuclear gene encoding the largest subunit of RNA polymerase I, comprises highly polymorphic intron 19 and nucleotide tag (Ntag) sequences. We analyzed these sequences in 42 accessions, which differed in ploidy, of Triticum–Aegilops and Hordeum species. The lengths of the intron 19 sequences were ca. 110?bp long in Triticum–Aegilops species, except in four Sitopsis species, Ae. longissima, Ae. searsii, Ae. sharonensis, Ae. speltoides, which had introns similar in length to those of Hordeum species, i.e., ca. 240?bp long. Phylogenetic analyses of the Ntag sequences showed that the four Sitopsis and remaining Triticum–Aegilops species were classified into two discrete Hordeum and Triticum clades, respectively. The A and D genome-specific Ntag sequences of polyploid wheats were highly homologous with those of T. urartu and Ae. tauschii, respectively. In Ae. bicornis, another Sitopsis species, two accessions had the short intron 19 and Triticum–type Ntag sequence, which were highly homologous with those of the B genome in polyploid wheats, whereas one accession contained the long intron 19 and Hordeum–type Ntag sequences. In contrast, partial sequence analyses revealed that the three accessions of Ae. bicornis shared highly homology to single-copy DMC1 and EF-G genes. The discrepancy between these results indicates that the Sitopsis species were probably established by hybrid speciation including ancient introgressive hybridization between progenitors of Triticum–Aegilops and Hordeum. Although many researchers have proposed Ae. speltoides as a candidate for the B genome donor, our data suggest the existence of diploid B genome species in the past that were responsible for the origin of both polyploid wheats and Sitopsis species, including Ae. speltoides.     

Identification of low-molecular weight glutenin subunits of wheat associated with bread-making quality

Although it is known that the compositions of low‐molecular weight glutenin subunits (LMW‐GSs) are important factors for bread‐making quality of wheat, it is still not clear which LMW‐GSs confer improved bread‐making quality and how those LMW‐GSs interact with high‐molecular weight (HMW) GSs. Using a hard red winter wheat line with good bread‐making quality and a Japanese wheat cultivar with poor quality as well as their progeny we identified LMW‐GSs associated with the bread‐making quality. One such LMW‐GS, KS2, which had a molecular weight of 42 kDa and was allelic to HS1, was associated with bread‐making quality. Furthermore, by using four recombinant inbred lines with different HMW‐GS and LMW‐GS combinations, KS2 and HMW‐GS 5+10 showed interaction effects on the bread‐making quality. Two‐dimensional polyacrylamide gel electrophoresis (Page) analysis showed that KS2 consists of two protein components and that HS1 is composed of three components. The N‐terminal amino acid sequences of the five components were identical to the most frequently analysed sequence of LMW glutenin components.     

Quantitative structure-activity relationships of the fungicidal methyl N-phenylcarbamates

For 69 methyl N-phenylcarbamates having various benzene ring substituents, the fungicidal activity was determined against Botrytis cinerea resistant to benzimidazole fungicides by the agar medium dilution method. The structure-activity relationships were analyzed quantitatively using such physicochemical substituent parameters as hydrophobic π, steric B₅, and HB (hydrogen bonding) with the Hansch-Fujita method (C. Hansch and T. Fujita, J. Amer. Chem. Soc.86, 1616 1964). The hydrophobicity of substituents was favorable to the acitivity. The effect was position-specific, the importance being in the order of ortho meta > para. The activity was related parabolically to the maximum width of one of the m-substituents having the larger dimension. The hydrogen acceptability of p-substituents enhanced the activity. The preventive activity of compounds against gray mold of cucumber caused by the resistant B. cinerea, which was determined by the foliar application in pot tests, was dependent on the magnitude of the fungicidal activity and hydrophobicity of the compounds as analyzed by the adaptive least-squares method.     

Presence of a Temperature Gradient Among Genital Tract Portions and the Thermal Changes Within These Portions Over the Estrous Cycle in Beef Cows

The aim of the present study was to describe the temperature of the different portions of the female genital tract and their relation to rectal temperature and to investigate the effect of steroid hormones profiles on these variables over the estrous cycle in cattle. Four nonpregnant Japanese Black cows were investigated daily over two successive estrous cycles using a digital thermometer with a long probe and rounded-end sensor to record the temperature of the rectum (RT), vagina (VT), cervix (CT), uterine body (UBT) and uterine horns (UHT). Blood samples were collected immediately before temperature recording to assay peripheral levels of progesterone (P₄) and estradiol-17β (E₂). Moreover, transrectal ultrasonography was carried out after temperature recording to monitor the ovulatory follicle and track ovulation. During the experiment, the ambient temperature and relative humidity were recorded for further calculation of the temperature humidity index (THI). The temperature within the genital tracts in these cows progressively increased towards the uterine horns from the vagina. The VT, CT, UBT and UHTs were significantly higher in association with peripheral P₄ concentrations greater than 4 ng/ml (mid-luteal phase) when compared with lower peripheral P₄ concentrations. The VT was more significantly (P<0.01) correlated to the CT, UBT and UHTs than RT. In conclusion, a temperature gradient was present among the vagina, cervix and uterus over the estrous cycle, and changes in peripheral P₄ concentrations were associated with the thermal variations within these portions. The VT could be more beneficial than RT in monitoring temperature of deeper portions of the female genital tract in bovine.     

Isolation of Mycobacterium spp. from milking buffaloes and cattle in Nepal

In Nepal, mycobacterial isolates obtained from the milk and feces of buffaloes and cattle that were positive for the single intradermal cervical tuberculin (SICT) tests were genetically identified. A total of 36 mycobacterial strains were isolated from 39% of the buffaloes (14 of 36) and 34% of the cattle (11 of 32). Of the 36 strains, 13 were identified as M. bovis, and these strains were isolated from 17% of the buffaloes (6 of 36) and 16% of the cattle (5 of 32). M. bovis was isolated from both the milk and feces of one buffalo and one cattle, the milk alone of three buffaloes and three cattle, and the feces alone of two buffaloes and one cattle. These results suggest that milking buffaloes and cattle infected with M. bovis exist in Nepal. The remaining 23 strains were atypical mycobacteria. A program for the elimination of bovine tuberculosis should be implemented as soon as possible, and the public health education and proper hygienic practices may be required.     

Changes in mRNA of immune factors expressed by milk somatic cells of Holstein cows with hypocalcemia after calving

Changes in immune factors expressed by milk somatic cells from Holstein cows with hypocalcemia after calving were investigated in this study. Fourteen multiparous Holstein cows after their 3rd or 4th calving in one farm were used. The cows were divided into 2 groups: 7 cows needing treatment due to onset of hypocalcemia (hypocalcemia group; age = 5.53 ± 0.27 years, parity = 3.14 ± 0.14) and 7 cows without health problems (control group; age = 5.88 ± 0.31 years, parity = 3.57 ± 0.26). Milk samples were collected aseptically using a cannula and mRNA of immune factors expressed by milk somatic cells were analyzed. Milk samples (50 mL) were collected from the right rear mammary gland of cows before milking at day 1 and weeks 1, 2, 4, and 8 after calving. All milk samples showed a negative reaction to the California Mastitis Test. Levels of relative interleukin (IL)-6 and cathelicidin in the hypocalcemia group were lower than those in the control group in weeks 1 to 8. A significant difference in relative IL-6 levels was found in week 4 (P < 0.05). These results suggest that levels of IL-6 expressed by milk somatic cells may be affected by hypocalcemia in dairy cows.     

Anthracnose of salt-wort (<Emphasis Type="Italic">Salsola komarovii</Emphasis>) caused by <Emphasis Type="Italic">Colletotrichum truncatum</Emphasis>

In July 2006, black rot was observed on the leaves of 4-leaf-stage seedlings of salt-wort (Salsola komarovii) in Yamagata Prefecture, Japan. We isolated two single-conidial isolates from the diseased leaves. Although colony appearance of the isolates was different from that of each other, both isolates were identified as Colletotrichum truncatum by morphology and molecular similarity. After inoculation of healthy salt-wort plants with the isolates, the isolates were reisolated from symptomatic plants. We thus propose a new disease, anthracnose of salt-wort.     

Fish meal replacement by soy protein from soymilk in the diets of red sea bream (Pagrus major)

Six isoenergetic diets were formulated as follows: fish meal (FM) 700 g kg^–1 (control, C), FM 300 g kg^–1 + soy protein concentrate 300 g kg^–1 (SPC), FM 300 g kg^–1 + enzyme‐treated SPC 300 g kg^–1 (ESC), FM 170 g kg^–1 + soy protein isolate 300 g kg^–1 (SPI), FM 160 g kg^–1 + enzyme‐treated SPI 300 g kg^–1 (ESI) and FM 150 g kg^–1 + conglycinin 300 g kg^–1(CG). Forty fish (3.9 g) were randomly distributed into each of eighteen 300‐L tanks, fed twice daily until satiation for 8 weeks. The final body weight, specific growth rate and condition factor did not show significant differences among the fish fed with diets C, SPC, ESC and ESI (p > .05). The survival was significantly lower in fish fed with diets SPI and CG. Feed efficiency was significantly higher in fish fed with diets SPC and C than in fish fed with other diets (p < .05). There were no significant differences in nutrients retention efficiencies in fish fed with diets C, SPC, ESC and ESI. A significantly higher phosphorus retention efficiency in fish fed with soymilk protein diets resulted in lower phosphorus discharge to the environment (p < .05). These results suggest that the soymilk proteins can comfortably replace 570–770 g FM kg^–1 diet of red sea bream juvenile, which will ensure significant ecological benefits through reducing phosphorus load to the environment.