Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis

The effect of intramammary injection of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF, 400 microg/10 mL) on quarter milk levels of chemiluminescence (CL) activity, and somatic cell count (SCC) and shedding pattern of Staphylococcus aureus was investigated. Ten Holstein cows, naturally infected with S. aureus were used, with either early-stage or late-stage subclinical mastitis. Injection of rboGM-CSF caused a remarkable increase in milk CL activity with a peak at 6 h after the cytokine injection in the early- and late-stage groups. In the early-stage group, milk SCC stayed around preinjection level at 6 h, rose significantly on days 1 and 2, and was followed by a smooth and significant decline to an under preinjection level (below 200 000 cells/mL) on day 7 postinjection. Alternatively, in the late-stage group, milk SCC rose significantly at 6 h after the cytokine injection and maintained high levels thereafter. The milk S. aureus count decreased drastically by the cytokine injection in the early-stage group. The bacterial count was moderately decreased in the late-stage group, but increased back to preinoculation levels on day 7 after the cytokine injection. The results suggest that the rboGM-CSF has a potential as a therapeutic agent for S. aureus infection causing subclinical mastitis of dairy cows, if the cytokine is applied at the initial stage of infection.     

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.     

Transformation of trifoliate orange with rice chitinase gene and the use of the transformed plant as a rootstock

RCC2, cDNA clone encoding rice class-I chitinase, was introduced into trifoliate orange (Poncirus trifoliata Raf.). Chitinase activity of the transformed plants was higher than that of the non-transformed plants. The introduced gene was not detected in the scions grafted onto the transformed plants. The effect of the introduction of foreign gene into rootstocks on the scions was discussed.     

Patterns of stable carbon and nitrogen isotopes in the baleen of common minke whale Balaenoptera acutorostrata from the western North Pacific

ABSTRACT:   Stable carbon and nitrogen isotope ratios were determined in the baleen plates of 17 common minke whales Balaenoptera acutorostrata from the north-western Pacific Ocean off Japan, as well as prey species (krill Euphausia pacifica , Japanese anchovy Engraulis japonicus and Pacific saury Cololabis saira ) collected in the stomach contents, to investigate the trophic relationship between the minke whales and their prey. A few δ¹⁵N-depleted peaks occurred along the length of baleen plates for 10 males irrespective of stomach content (anchovies and sauries). Similar δ¹⁵N-depleted peaks were also found for one female and two immature individuals. It was likely that these δ¹⁵N-depleted peaks formed in early summer. The stable nitrogen isotope ratio (δ¹⁵N) values in Pacific saury (9.3 ± 1.4‰) did not differ significantly from that in Japanese anchovy (8.8 ± 0.9‰). In contrast, δ¹⁵N in krill (7.2 ± 0.5‰ in July and 8.0 ± 0.2‰ in September) were significantly lower than in the Pacific saury. Thus, these peaks may reflect the dietary change from krill to fishes in the feeding migration of the whales. Growth rate of the baleen plate was estimated to be 129 mm/y, and it appeared that a dietary record of about 1.4 years remained in the baleen plate. For two immature whales, the maximum value of δ¹⁵N occurred at the tip of baleen. This δ¹⁵N enrichment may possibly be useful for discriminating weanlings and older whales.     

Isolation of influenza A viruses from migratory waterfowl in San-in District, western Japan in the winter of 1983-1984

Certain species of winter migratory waterfowl in San-in District, western Japan, were surveyed for influenza virus from November 1983 to March 1984. Faeces of the waterfowl were collected regularly at five stations. Eleven influenza A viruses including H5N3 and H10N4 subtypes were isolated from 450 faecal samples from whistling swans, 28 viruses including H2N2 and H10N4 subtypes were isolated from 362 faecal samples from pintails; and subtype H13N6 was isolated from 240 faecal samples of black-tailed gulls.     

Isolation and culture of rabbit primordial germ cells

Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells.     

Habitat differentiation between sei (Balaenoptera borealis) and Bryde's whales (B. brydei) in the western North Pacific

Two closely related baleen whale species, sei and Bryde's whales, in the western North Pacific were studied to identify differences in habitat use. Data were obtained from May to August 2004 and 2005. This study examined the relationship between oceanographic features derived from satellite data and the distribution of sei and Bryde's whales using basic statistics. We investigated oceanographic features including sea surface temperature (SST), sea surface chlorophyll a (Chl‐a), sea surface height anomalies (SSHAs), and depth of the habitat. These two whale species used habitats with different SST, Chl‐a, and SSHA ranges. The 0.25 mg m^?3 Chl‐a contour (similar to the definition of the Transition Zone Chlorophyll Front) was a good indicator that separated the habitats of sei and Bryde's whales. Then generalized linear models were used to model the probabilities that the whale species would be present in a habitat and to estimate their habitat distribution throughout the study area as a function of environmental variables. The potential habitats of the two species were clearly divided, and the boundary moved north with seasonal progression. The habitat partitioning results indicated that SST contributed to the patterns of habitat‐use and might reflect differences in prey species between the two whales. This study showed that the habitats of the sei and Bryde's whales were clearly divided and their potential habitat‐use changed seasonally.