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.     

Effect of timing and type of supplementary grain on herbage intake,nitrogen utilization and milk production in dairy cows grazed on perennial ryegrass pasture from evening to morning

The present study aimed to clarify the effect of timing and type of supplementary grain in grazing dairy cows on herbage dry matter intake (HDMI), nitrogen utilization and milk production. Eight lactating cows were allowed to graze from evening to morning during three seasonal periods (spring, summer, autumn). They were randomly allocated to four treatments (timing: pre‐ (Pre) or post‐grazing (Post), for large grain allotments consisting of 75% of daily grain offered; grain type: barley or corn) in 4 × 4 Latin square designs in each period. In the spring period, HDMI was greater for cows fed corn than those fed barley (P = 0.005), whereas cows in the Pre treatment had a similar HDMI, higher (P = 0.049) urinary purine derivative concentration and greater (P = 0.004) milk yield compared with cows in the Post treatment. In the summer and autumn periods, timing treatments did not affect HDMI, nitrogen utilization or milk production, but cows supplemented with barley had higher urinary purine derivatives concentration (P < 0.05) and milk yield (P < 0.05) compared with those supplemented with corn. The results indicate that large grain allotments immediately before evening grazing during early grazing seasons increased ruminal microbial protein synthesis and milk production without reducing HDMI regardless of grain type.     

Characterization of the cathelicidin cluster in the Japanese quail (Coturnix japonica)

The Japanese quail has several advantages as a low‐fat meat bird with high immunity against diseases. Cathelicidins (CATHs) are antimicrobial peptides that play an important role in innate immunity. The aim of this study was to characterize the CATH cluster in the Japanese quail (Coturnix japonica). The Japanese quail CATH (CjCATH) cluster, contains four CATH genes, as in the chicken. The coding sequences of CjCATHs exhibited >85.3% identity to chicken CATHs. The predicted amino acid sequences of the four CjCATH genes contained the cathelin‐like domain characteristic of CATH proteins. Polymorphisms were detected in the open reading frames (ORFs) of all CjCATH sequences. Two amino acid substitutions were observed in the antimicrobial region of the mature peptide of CjCATH2, and predicted to influence peptide function. CjCATH1 is expressed in lung, heart, bone marrow and bursa of Fabricius (BF). CjCATH2 is expressed in bone marrow. CjCATH3 is expressed in lung, heart, bone marrow, BF, tongue and duodenum. CjCATHB1 is expressed in bone marrow and BF. This study is the first to characterize CATH genes in the Japanese quail, and identifies novel antimicrobial peptide sequences belonging to the cathelicidin family, which may play a role in immunity in this species.     

Alteration of somatostatin receptor 2 expression in canine mammary gland tumor

Somatostatin receptor 2 (SSTR2) is a negative regulator of cell proliferation in human breast cancer. Since there is little information about SSTR2 in canine mammary gland tumor (MGT), we clarified its distribution and expression level in normal mammary gland, benign MGT and malignant MGT. SSTR2 expression determined by immunohistochemical staining was observed in the cytoplasm of luminal epithelial cells. The intensity was negatively correlated with malignancy: normal tissues and some of the benign tumors had the highest levels, while the malignant tumors had little or no SSTR2 expression. As for the Western blotting, SSTR2 protein level in benign tumors was significantly lower than the normal mammary gland. On the other hand, SSTR2 protein levels in two of three malignant tumors were higher than the other groups. These results suggest that SSTR2 expression alters according to the malignancy of canine MGT.     

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.     

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.     

Carbon source and trophic position of pelagic fish in coastal waters of south-eastern Izu Peninsula,Japan, identified by stable isotope analysis

ABSTRACT:   It is important to clarify trophic dynamics in marine ecosystems for management of the fishing ground. Organic carbon sources and trophic position of pelagic fishes in the coastal waters of the south-eastern Izu Peninsula, Japan, were examined on the basis of carbon and nitrogen stable isotope distributions. The δ¹³C of the fishes was mostly distributed from −19 to −16‰ for nektonic fishes (13 species of adults and immatures) and planktonic fishes (10 species of larvae and juveniles), close to the δ¹³C values of particulate organic matter and planktonic decapods. These δ¹³C signatures for the inhabitants of the water column were in contrast with the high δ¹³C values (mainly −16 to −13‰) for demersal fishes of Scorpaeniformes and benthic polychaetes collected in the surf zone. These results indicate that nektonic and planktonic fishes depend on phytoplankton for carbon supply. The δ¹⁵N signatures suggest that the trophic position ranged 3.1–4.5 for the nektonic fishes and 2.9–3.7 for the planktonic fishes, premised on trophic level 3 for larval Japanese anchovy Engraulis japonicus . Thus, planktivorous fishes should be mainly assigned to trophic levels 3 and 4 in this area.     

Characterization of vitamin B12 compounds from commercially available fish roe products

Although fish roes (or the egg-laden ovary) are seafood products with high nutritional value and are considered abundant in vitamins including vitamin B₁₂, nevertheless, the detailed properties of vitamin B₁₂ have not been fully elucidated in fish roe products. Vitamin B₁₂ content was determined using C18-reversed-phase high-performance liquid chromatography after purification of samples from immunoaffinity columns. Salmon egg-laden ovary products (sujiko), salmon roe products (ikura), dried mullet roe products (bottarga), and white sturgeon roe products (caviar) were found to contain substantial levels of vitamin B₁₂ (more than approximately 15 µg/100 g wet weight). Interestingly, high levels of vitamin B₁₂ per 100 g wet weight have been measured in pollack roe products (tarako) and flying fish roe products (tobiko). Liquid chromatography–electrospray-ionization/tandem mass spectrometry analysis revealed that vitamin B₁₂ was the predominant corrinoid in the evaluated fish roe products examined, while no inactive corrinoid compounds were identified. These results suggest that commercially available fish roe products could be an important source of vitamin B₁₂ for use as a dietary product for humans.

     

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.