Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

Discrimination of “grazing milk” using milk fatty acid profile in the grassland dairy area in Hokkaido

Milk produced by the grazing system, referred to as “grazing milk” contains many components required for human health. The milk fatty acid (FA) profile is strongly associated with the diet on the farms. In the present study, based on the FA profile of farmer's bulk milk, we determined how to discriminate between milk produced on grazing and on a confinement system. A field survey was conducted four times (grazing and confinement season) in the Konsen (29 farms) and Okhotsk (25 farms) area in Hokkaido. Farmer's bulk milk samples and details of feeding management were collected and the FA profile of milk was measured. Milk produced during the grazing season contained less C16:0 and cis‐9 C16:0, and more C18:0, cis‐9 C18:1, trans‐11 C18:1, cis‐9,12 C18:2, cis‐9,trans‐11 C18:2 and cis‐9,12,15 C18:3 than milk produced during the confinement season. Discrimination analysis using 16 FA revealed that almost all milk samples were discriminated correctly (confinement season: 90% correct and 10% borderline, grazing season: 88% correct, 9% borderline and 3% incorrect). For farmers that were categorized incorrectly and were considered borderline in the grazing season, the dependency on pasture was low compared with that for farmers correctly discriminated. Therefore, to claim “grazing milk”, a high dependency on pasture is required for grazing dairy farmers.     

Relationship of rumen fill and fermentation to diurnal and seasonal variation of herbage intake in dairy cows grazed on perennial ryegrass pasture

To clarify the effect of digesta weight in the reticulorumen on diurnal and seasonal fluctuations in herbage intake, six ruminally cannulated, non‐lactating dairy cows were grazed on perennial ryegrass/white clover pasture during morning and evening sessions in spring and autumn. The digesta weight of fresh matter, dry matter (DM) and fiber in the reticulorumen at the beginning and the end of each grazing session was lower in spring than in autumn (P < 0.01). Although the digesta weight was similar between the sessions at the beginning of grazing, it was greater for the evening than for the morning at the end of grazing (P < 0.01). The large particles proportion in the digesta was lower for the morning than the evening (P < 0.01), and it was lower in spring than in autumn (P < 0.01). The concentrations of volatile fatty acids in rumen fluid were generally higher in spring compared with autumn. The herbage DM intake during the evening was greater compared with the morning in both seasons (P < 0.01). However, there was no difference in herbage DM intake between seasons. The results showed that the rumen digesta fill was not the sole factor explaining diurnal and seasonal variation of herbage intake in grazing dairy cows.     

Herbage intake and ruminal digestion of dairy cows grazed on perennial ryegrass pasture either in the morning or evening

This study aimed to clarify diurnal fluctuations of herbage intake, ruminal fermentation of herbage carbohydrates and proteins, and digesta particulate weight in the rumen of grazing dairy cows. Six ruminally cannulated, non‐lactating dairy cows were grazed on perennial ryegrass/white clover pasture either in the morning (04.00 to 08.00 hours) or the evening (16.00 to 20.00 hours). Cows grazed in the evening spent more time (P < 0.01) and consumed more herbage (P < 0.01) compared with cows grazed in the morning. Higher (P < 0.05) daily mean concentrations of total volatile fatty acid, propionate and n‐butyrate in rumen fluid were observed for cows grazed in the evening compared with cows grazed in the morning. Although cows grazed in the evening ingested more crude protein compared with cows grazed in the morning, no significant difference in NH₃‐N concentration in rumen fluid was observed between them. The ratio of purine‐derivative concentration to creatinine concentrations was higher (P < 0.01) in the urine of cows grazed in the evening than in cows grazed in the morning. These results clearly indicated that evening grazing was advantageous for dairy cows compared with morning grazing, in terms of ruminal fermentable energy intake and nitrogen utilization efficiency.     

Androgen induces production of male effect pheromone in female goats

Previously we showed that the primer pheromone responsible for the "male effect" was produced in specific skin regions of castrated male goats by androgen treatments. In the present study, we examined whether androgen can also induce production of the male effect pheromone in female goats. Capsules containing dihydrotestosterone (DHT) or testosterone (T) were subcutaneously implanted into six ovariectomized (OVX) goats for 28 days. Small skin samples were collected from the head and rump regions, and the pheromone activity of their ether extracts was examined using a bioassay that monitors the electrophysiological manifestation of the hypothalamic gonadotropin-releasing hormone pulse generator as multiple-unit activity. Behaviors of OVX goats towards ovary-intact estrous goats were also examined before and at the end of DHT or T treatment. Before androgen treatment, neither the head nor rump skin samples in OVX goats showed pheromone activity. DHT treatment induced pheromone activity in the head skin sample of six OVX goats and in the rump skin sample of two OVX goats. Similar results were obtained by T treatment. In addition, OVX goats treated with T showed masculine-type sexual behaviors such as courtship and mounting behaviors towards the estrous goats. These results demonstrate that androgen is capable of inducing primer pheromone activity in the female and suggest that the synthesis pathway of the male effect pheromone exists in both sexes in the goat.     

Improvement of preservation method for ammonia-oxidizing bacteria by freeze-drying

Long-term preservation of ammonia-oxidizing bacteria had been achieved until now by using the silica gel-freezing method. Recently, the freeze-drying method for the preservation of microorganisms has been widely used, and has been applied to the preservation of ammonia-oxidizing bacteria. The 10% skim milk-l% pyruvate or glutamate ratio was optimum as a dispersion-medium, and the preservation temperature of the freeze-dried cells was effective at -80°C. This method of preservation was found to be considerably more effective than the silica gel-freezing method.     

Characterization of Pilot Scale Processed Canadian Organic Cranberry (Vaccinium macrocarpon) and Blueberry (Vaccinium angustifolium) Juice Pressing Residues and Phenolic-Enriched Extractives

This work examined the processing yields, physico-chemical characteristics, and bioactive properties of Canadian organic cranberry and blueberry pomace and phenolic-enriched extractives. The ethanol extracts from each berry contained three to four times the soluble solids and sugar contents compared to the pomace. The lipid content of organic cranberry pomace (4.44%), cranberry ethanol extractives (3.86%), and blueberry ethanol extractives (4.44%) were significantly lower than the lipid content of the organic blueberry pomace (5.42%). The organic blueberry pomace showed higher protein content (8.41%) than organic cranberry pomace (5.76%); however, both organic cranberry and blueberry ethanol extracts showed similarly low protein contents. Significant differences were observed in the mineral and elemental contents of both pomaces and their extractives. Total phenolics, tartaric esters, flavonols, anthocyanins, and antioxidant activities were increased by two to three times in both organic cranberry and blueberry ethanol extracts compared to their respective pomaces. The major anthocyanins detected in organic blueberry pomace and ethanol extracts were peonidin 3-glucoside, malvidin 3-galactoside, malvidin 3-glucoside, and cyanidin 3-arabinoside. Peonidin 3-galactoside, cyanidin 3-galactoside, cyanidin 3-arabinoside, and peonidin 3-arabinside were the major anthocyanins in the organic cranberry and ethanol extacts. Results indicated that cranberry and blueberry pomace could be used to develop bioactives with antioxidant activity for potential health benefit and/or bioenergy production.     

Effects of pre-feeding of a corn silage-based supplement on the feed intake, milk production and nitrogen utilization of grazing dairy cows

Four Holstein cows were used to determine the effect of timing of the feeding of a corn silage (CS)‐based supplement on the feed intake, milk production and nitrogen utilization of grazing dairy cows. The cows were fed the supplement 2 h before grazing (pre‐grazing) or immediately after grazing (post‐grazing). Cows were grazed for 5 h per day under a rotational grazing system. There was no difference in the herbage and total feed intake between treatments. The milk protein yield for pre‐grazing tended to be higher than that for post‐grazing, whereas the milk yield did not differ between treatments. The total nitrogen intake for pre‐grazing tended to be higher than that for post‐grazing (P = 0.06). There was no difference in the urinary nitrogen output between treatments, whereas the proportion of urinary nitrogen output : total nitrogen intake for pre‐grazing tended to be lower than that for post‐grazing (P = 0.06). The milk nitrogen output and nitrogen retention for pre‐grazing tended to be higher than that for post‐grazing (milk nitrogen, P = 0.06; nitrogen retention, P = 0.05). Nitrogen utilization of grazing dairy cows was improved by feeding a CS‐based supplement before grazing.     

Cycas necrotic stunt virus isolated from gladiolus plants in Japan

An undescribed spherical virus ca. 30 nm in diameter was isolated from gladiolus (Gladiolus spp.) plants in Japan. The virus had a moderate host range within eight families. Purified virus preparations contained two large RNA components and one coat protein with mobility similar to Cycas necrotic stunt virus (CNSV) from cycas (Cycas revolute). The virus was serologically closely related to CNSV. Its nucleotide sequence of the coat protein gene had 89% common identity with that of CNSV. These results indicated that the virus isolated from gladiolus is a new strain of CNSV. The nucleotide sequence data reported are available in the DDBJ/EMBL/Gen Bank databases under the accession number AB237656.