Characterization of Aerococcus viridans isolated from milk samples from cows with mastitis and manure samples

Thirty-eight Aerococcus viridans isolates were obtained from milk from 478 cows with clinical mastitis in a farm during the periods between November 2011 and February 2012, and between December 2012 and March 2013. Additional isolates were obtained from processed manure (a mixture of composted manure, straw and hydrated lime) and bedding materials. The processed manure was later used to cover the floor of the stalls in barns as bedding materials. The temperatures recorded in the composted and processed manure were not as high as those generally observed during satisfactory composting. To reveal the association of A. viridans in manure-related products with intramammary infection in cows, isolates were characterized by their DNA fragment patterns as determined by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing. Isolates obtained from milk, processed manure and bedding materials had identical DNA fragment patterns. Antimicrobial susceptibilities were determined for 29 isolates from milk, processed manure and bedding materials. Of these, 26 (89.7%) were resistant to clindamycin, whereas virtually all the isolates were susceptible to 12 other antimicrobials including cefalosporins that have been used to treat bovine mastitis in Japan. In vitro, three A. viridans isolates from milk and an isolate from processed manure survived for 3 hr in Good’s buffer (pH 9) at high temperature (50°C). The results suggest that the processed manure and bedding materials in this farm were possible sources of A. viridans that caused infection in the cows with mastitis.     

Weak activity of UDP-glucuronosyltransferase toward Bisphenol analogs in mouse perinatal development

Bisphenol A (BPA) is a widely used industrial chemical that disrupts endocrine function. BPA is an endocrine disrupting chemical (EDC) that has been demonstrated to affect reproductive organ development, brain development, metabolic disease and post-natal behavior. Accordingly, Bisphenol analogs, Bisphenol F (BPF, bis (4-hydroxyphenyl) methane) and Bisphenol AF (BPAF, 4,4-hexafluoroisopropylidene) diphenol) are used as replacements for BPA. BPA is mainly metabolized by UDP-glucuronosyltransferase (UGT), UGT2B1, but this effective metabolizing system is weak in the fetus. In the present study, we demonstrated that hepatic UGT activity toward BPAF was very weak, in comparison with BPA and BPF, in the fetus, pups and dams. Conversely, hepatic UGT activity toward BPF was very weak in the fetus and newborn pups, and was increased to the same level as BPA post-partum. In conclusion, BPAF possibly tends to accumulate in the fetus, because of weak metabolism during the perinatal period, suggesting that the metabolism of individual Bisphenol analogs requires assessment to properly gauge their risks.     

Pathogenicity of emerging Japanese type 1 porcine reproductive and respiratory syndrome virus in experimentally infected pigs

To clarify the pathogenicity of Japanese type 1 porcine reproductive and respiratory syndrome virus (PRRSV) isolate in experimentally infected pigs, we evaluated clinical signs and monitored viremia for 21 days post-inoculation (dpi). Lungs were mottled, tanned and reddish in appearance; had lesions predominantly in the cranial, middle and accessory lobes; and failed to collapse at 10 dpi. Although microscopic lesions of lungs were reproduced using the Japanese emerging type 1 PRRSV isolate under experimental conditions, no significant differences were noted between the challenge and control groups regarding mean rectal temperature and daily weight gain. These results provide useful insights into the limited pathogenicity of single infection with the Japanese type 1 PRRSV isolate in piglets, which differ from findings in reported field cases.     

Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.     

Mitigation of nitrous oxide (N2O) emission from swine wastewater treatment in an aerobic bioreactor packed with carbon fibers

Mitigation of nitrous oxide (N₂O) emission from swine wastewater treatment was demonstrated in an aerobic bioreactor packed with carbon fibers (CF reactor). The CF reactor had a demonstrated advantage in mitigating N₂O emission and avoiding NOx (NO₃ + NO₂) accumulation. The N₂O emission factor was 0.0003 g N₂O‐N/gTN‐load in the CF bioreactor compared to 0.03 gN₂O‐N/gTN‐load in an activated sludge reactor (AS reactor). N₂O and CH₄ emissions from the CF reactor were 42 g‐CO₂ eq/m³/day, while those from the AS reactor were 725 g‐CO₂ eq/m³/day. The dissolved inorganic nitrogen (DIN) in the CF reactor removed an average of 156 mg/L of the NH₄‐N, and accumulated an average of 14 mg/L of the NO₃‐N. In contrast, the DIN in the AS reactor removed an average 144 mg/L of the NH₄‐N and accumulated an average 183 mg/L of the NO₃‐N. NO₂‐N was almost undetectable in both reactors.     

Preparation, characterization, and antioxidative effects of oligomeric proanthocyanidin-L-cysteine complexes

Controlled acid-catalyzed degradation of proanthocyanidin polymers in grape seeds together with L-cysteine led to oligomeric proanthocyanidin-L-cysteine complexes along with monomeric flavan-3-ol derivatives being isolated, and their structures were confirmed on the basis of spectroscopic data and by chemical means. In addition, comparative studies on the antioxidative and survival effects of oligomeric proanthocyanidin-L-cysteine complexes and proanthocyanidin polymers were performed. The oligomeric proanthocyanidin-L-cysteine complexes showed higher bioavailability and antioxidant capacity and enhanced survival time in the animal test groups. In addition, it is suggested that the oligomeric complexes may help to prevent oxidative stress and may reduce free radical production.     

Specific detection of potentially allergenic kiwifruit in foods using polymerase chain reaction

Kiwifruit (Actinidia deliciosa and Actinidia chinensis) is allergenic to sensitive patients, and, under Japanese regulations, it is one of the food items that are recommended to be declared on food labeling as much as possible. To develop PCR-based methods for the detection of trace amounts of kiwifruit in foods, two primer pairs targeting the ITS-1 region of the Actinidia spp. were designed using PCR simulation software. On the basis of the known distribution of a major kiwifruit allergen (actinidin) within the Actinidia spp., as well as of reports on clinical and immunological cross-reactivities, one of the primer pairs was designed to detect all Actinidia spp. and the other to detect commercially grown Actinidia spp. (i.e., kiwifruit, Actinidia arguta, and their interspecific hybrids) except for Actinidia polygama. The specificity of the developed methods using the designed primer pairs was verified by performing PCR experiments on 8 Actinidia spp. and 26 other plants including fruits. The methods were considered to be specific enough to yield target-size products only from the target Actinidia spp. and to detect no target-size products from nontarget species. The methods were sensitive enough to detect 5-50 fg of Actinidia spp. DNA spiked in 50 ng of salmon testis DNA used as a carrier (1-10 ppm of kiwifruit DNA) and 1700 ppm (w/w) of fresh kiwifruit puree spiked in a commercial plain yogurt (corresponding to ca. 10 ppm of kiwifruit protein). These methods would be expected to be useful in the detection of hidden kiwifruit and its related species in processed foods.     

Advanced broodstock diets for the mangrove red snapper and a potential importance of arachidonic acid in eggs and fry

Mangrove red snapper fed advanced broodstock diets containing squid meal and squid oil exhibited higher hatching rates, cumulative survival and survival activity index than those fed a basal diet or a basal diet supplemented with mixture of antioxidants. On the other hand, fatty acid analyses of ovaries and fry of wild fish and eggs and larvae of broodstock fed raw fish revealed high arachidonic acid (ARA) and docosahexaenoic acid (DHA) levels and relatively lower eicosapentaenoic acid (EPA) levels consequently showing high ARA/EPA and DHA/EPA ratios compared to cold water species. This suggests that ARA may be nutritionally more important for egg and larval development and survival in tropical marine fish and its supplementation in broodstock diets may enhance reproductive performance of mangrove red snapper.     

The duality of teleost gonadotropins

The duality of salmon gonadotropins has been proved by biochemical, biological, and immunological characterization of two chemically distinc gonadotropins. GTH I and GTH II were equipotent in stimulating estradiol production, whereas GTH II appears to be more potent in stimulating maturational steroid synthesis. The ratio of plasma levels and pituitary contents of GTHs and the secretory control by a GnRH suggest that GTH I is the predominant GTH during vitellogenesis and early stages of spermatogenesis in salmonids, whereas GTH II is predominant at the time of spermiation and ovulation. GTH I and GTH II are found in distinctly separate cells. In trout, GTH I is expressed first in ontogeny, whereas GTH II cells appear coincident with the onset of spermatogenesis and vitellogenesis, and increase dramatically at the time of final reproductive maturation. Comparison of the amino acid sequences of polypeptides and the base sequences of cDNA revealed that salmon GTH I β is more similar to bovine FSHβ than bovine LHβ and salmon GTH II β shows higher homology to bovine LHβ than to bovine FSHβ. The existence of two pituitary gonadotropins in teleosts as well as tetrapods suggests that the divergence of the GTH gene took place earlier than the time of divergence of teleosts from the main line of evolution leading to tetrapods.