Distribution and Fate of Organotin Compounds in Japanese Coastal Waters

In order to elucidate the details of both the distribution and fate of organotin compounds (OTs) in the costal ecosystem, the concentrations of butyltin compounds (BTs) and phenyltin compounds (PTs) were determined in seawater, sediment and blue mussels Mytilus galloprovincialis collected in Maizuru Bay, Japan. The concentrations of tributyltin (TBT) in seawater, sediments and mussels ranged from 3.9 to 27 ng l^?1, from 1.2 to 19 ng g^?1 dry wt and from 0.77 to 11 ng g^?1 wet wt, respectively. Although the levels of TBT in seawater, sediments and mussels from Maizuru Bay were lower than those reported previously at other sites in Japan, the levels can still be toxic to susceptible organisms. Trace amounts of PTs were also found in seawater, sediment and mussel samples, indicating that there is a slight input of triphenyltin (TPT) into the seawater in the bay at present. The highest TBT concentration in seawater among all sites measured in the bay was found near a glass factory, and the lowest concentrations were observed at the center of the bay. The highest concentration of TBT in sediment was detected near a shipyard. In mussels, a high concentration of TBT was detected near a cement plant and timberyard. These results indicate that the major sources of contamination of OTs in the bay are considered to be from those facilities and ships. The proportion of TBT in seawater and mussels showed that, of total BTs, it was the predominant compound at most sites. These results suggested that there is a continuous input of TBT into the seawater and thereafter an accumulation of TBT in mussels due to their low metabolic capacity to degrade TBT. In sediment, the proportion of monobutyltin (MBT) was the highest of BTs at most sites. This could reflect a previous contamination by TBT used before the regulation of its usage in the bay.     

Fatty acid‐binding protein expression in the gastrointestinal tract of calves and cows

Fatty acid‐binding protein (FABP) has high affinity for long‐chain fatty acids and appears to participate in the metabolism and intracellular transport of lipids. Liver‐ and intestinal‐type FABP (L‐FABP and I‐FABP, respectively) are expressed in the small intestine. However, in the gastrointestinal tract of ruminants, expression and localization of FABPs are unknown. In this study, we investigated the expression of I‐FABP and L‐FABP in the gastrointestinal tract of cattle. I‐ and L‐FABP had higher messenger RNA (mRNA) and protein expression levels in the duodenum and jejunum relatively to other gastrointestinal regions in both calves and cows. Furthermore, L‐FABP mRNA and protein expression were high in the colon. Both these protein types were confirmed to be in the cytosol of jejunal epithelial cells, where they were found in the villi rather than in the crypts. We concluded that duodenal and jejunal FABPs might be involved in the metabolism of fatty acids mainly in epithelial cells in cattle.     

Comparison of the insulin reaction of peripheral blood T cells between healthy Holstein dairy cows and JB during the periparturient period

To compare the changes in the insulin reaction of Holstein dairy cows and Japanese Black cows (JB) during the periparturient period, the insulin resistance test in vivo and lymphocytes proliferation with insulin in vitro were performed. Ten healthy Holstein dairy cows (Holstein group) and 10 healthy JB cows (JB group) used in this study were observed on days 60, 40, and 20 before calving and days 7 and 20 after calving. In insulin resistance reaction in vivo and in vitro, a low insulin-stimulated glucose disposal rate and lymphocyte proliferation with insulin were observed in the Holstein group compared with the JB group during the experimental period. An analysis of the lymphocytes cultured with insulin showed that the percentage of CD4+CD45R- T cells in the Holstein group was significantly lower than that of the JB group before day 20. These findings indicate that T cells reaction to insulin in healthy periparturient Holstein cows is lower than that in Japanese Black.     

Effects of serotonin injected into the third ventricle on prolactin and growth hormone secretion in Holstein steers

In order to clarify the role of serotonin (5-HT) in the regulation of pituitary hormones, the effects of 5-HT injected into the third ventricle (3V) on prolactin (PRL) and growth hormone (GH) release were investigated in Holstein steers. A chronic cannula was implanted in 3V by stereotaxic surgery under general anesthesia. After sufficient recovery from surgery, 5-HT (0, 0.1, 1.0, 2.0 mg) was injected into via the cannula and blood samples were collected over 4 h. Plasma PRL and GH concentrations were determined by radioimmunoassay. PRL release was significantly stimulated by the injection of 5-HT. The increase in PRL was observed at 20 min after the injection at three doses and the highest dose (2.0 mg) was the most effective in stimulating PRL release. The injection of 5-HT into 3V, at all doses tested, did not alter GH release significantly. Our results suggest that 5-HT is involved in the regulation of PRL release partly through the hypothalamus in cattle.     

Effects of active egg white product/ Clostridium butyricum Miyairi 588 additive on peripheral leukocyte populations in periparturient dairy cows

The leukocyte populations of periparturient dairy cows were analyzed after administration of active egg white/Clostridium butyricum Miyairi additive. Sixty-eight Holstein milking cows were divided into 3 groups. Group A was administered active egg white product (AEWP)/Clostridium butyricum Miyairi 588 (Miyairi 588) additive (n=23). Group B was administered Miyairi 588 only (n=23), and Group C was the control group (n=22). The challenged groups were administered 100 g of AEWP + Miyairi 588, or Miyairi 588 alone, daily for 60 days from 1 month before until 1 month after paturition. Blood samples were collected from all groups three times (1 month before, 1 week after and 1 month after parturition) for analysis of the peripheral leukocyte population. The results showed significantly higher numbers of CD4+ cells in Group A compared with Group C 1 week after paturition. AEWP/Miyairi 588 additive may enhance the number of CD4+ T cells in periparturient dairy cows.     

Intracerebroventricularly administered oxytocin attenuated cortisol secretion, but not behavioral responses, during isolation in Holstein steers

In rodents, intracerebroventricular oxytocin administration attenuated hypothalamo-pituitary-adrenal (HPA) responses and anxiety behavior during stress. We examined the effects of intracerebroventricular injection of oxytocin on isolation-induced stress responses in cattle. In a methodological test, we determined the dosage of oxytocin applied in a main test which did not induce an increase in plasma cortisol concentration or stereotyped behaviors. In a main test, 5 steers aged from 199 to 250 days were assigned to the following three treatments randomly: T1, no isolation after injection of 200 microl of artificial cerebrospinal fluid (aCSF); T2, isolation after aCSF injection; and T3, isolation after 0.5 microg of oxytocin in 200 microl aCSF injection. The isolation was conducted by leaving the experimental steer alone in its stall for one hour while its peers were taken outside. In T2, the isolation induced a rapid increase in plasma cortisol concentration. The maximum %-changes from the pre-isolation value were significantly attenuated by oxytocin injection (T2 vs. T3, p<0.05). The isolation also induced an increase in the frequency (number of occurrences/1 hr isolation) of vocalizations and body orientation changes, and a decrease in the percentage of time spent lying and ruminating. The effect of oxytocin on these behavioral responses to isolation was not apparent. These results indicate that intracerebroventricularly injected oxytocin at low dose attenuated the cortisol response to isolation in steers while the effect on behavior was very small in this experimental condition.     

Decrease in bovine CD14 positive cells in colostrum is associated with the incidence of mastitis after calving

During the postpartum period there is a high incidence of mastitis in dairy cows. The reason for this increased risk of mastitis still remains unclear. Since leukocytes in colostrum have an important role in preventing the onset of mastitis, we investigated the leukocyte populations, which express CD4, CD8, CD14, CD21 or WC1, in colostrum as well as in blood obtained from 14 Holstein cows. Eight cows developed mastitis within a week after calving and the other 6 remained healthy. The percentage of CD14+ cells in colostrum was significantly lower in mastitic cows than in healthy cows. There were no significant differences in other marker positive cells either in the colostrum or in the blood. The CD14+ cells in colostrum play an important role of defense against invading microorganisms in the mammary glands. Our results suggested that the lower percentage of CD14+ cells in colostrum might predict the incidence of mastitis in the following period.     

Generation of monoclonal antibodies specific for ORF68 of koi herpesvirus

Outbreaks of koi herpesvirus (KHV) infection in carp are still a serious problem worldwide. KHV is closely related to other two cyprinid herpesviruses, pox herpesvirus (CHV) and haematopoietic necrosis herpesvirus (CyHV-2) in goldfish. In this study, two major KHV antigenic proteins (ORF62 and ORF68) were identified by immunoscreening using a KHV-specific polyclonal antibody, and then monoclonal antibodies were generated for immunodiagnostic studies. After screening hybridoma cells, one mAb against ORF68 (mAb-7C6) was obtained but no mAbs against ORF62. mAb-7C6 specifically reacted with a lysate of KHV-infected koi fin cells (KF-1 cells) but not with lysates of CHV- or CyHV-2-infected KF-1 cells in an immuno-blotting analysis. Similar results were shown in the following tests: (1) a indirect fluorescent antibody test using infected KF-1 cells and (2) an immunohistochemical investigation by fast red stain (infected liver) or FITC detection (infected spleen). These results suggested that mAb-7C6 specifically reacts with KHV ORF68 protein.     

Occurrence of two distinct Theileria lineages in sika deer (Cervus nippon) of Iwate Prefecture, Japan

In the present study, we tried to detect protozoan blood parasites from the liver or blood of 141 Japanese sika deer (Cervus nippon) in Iwate Prefecture of Japan by polymerase chain reaction. Approximately 500-bp amplicons were obtained in 76 (53.9%) of 141 samples by amplification for V4 hyper-variable regions of the 18S rRNA gene, and the amplicons were considered to be DNA of Theileria species. The complete nucleotide sequences (1701-bp) of the 18S rRNA gene were determined in 25 samples and were divided into 8 genotypes that were phylogenetically separated into two distinct lineages showing a monophyletic relation. The two lineages of Theileria were detected in different rates (12 and 88%) from sika deer in Iwate Prefecture.     

Effects of intravenous tryptophan infusion on thermoregulation in steers exposed to acute heat stress

This study was conducted to investigate the effect of tryptophan (TRP) supply on the thermoregulatory responses via brain serotonin (5‐HT) in cattle. In period 1, 12 Holstein steers were kept under a constant room temperature (22°C) and were administered the intravenous (i.v.) infusion of saline or TRP (38.5 mg/kg/2 h). Changes in rectal temperature (RT), 5‐HT concentration in the cerebrospinal fluid (CSF), and other factors involved in thermoregulation were measured. In period 2, the steers received the same treatments as in period 1; however, the room temperature was elevated from 22°C to 33°C during i.v. infusion and maintained at 33°C for 3 h. 5‐HT concentration in CSF increased following TRP infusion in both periods, and RT significantly decreased following TRP infusion only in period 2. The effect of TRP on respiration rate and plasma prolactin and total triiodothyronine concentrations was not significant. These results suggest that increase in TRP supply can attenuate increase in RT in response to acute heat stress through the increase in brain 5‐HT, followed by presumable increase in evaporative heat loss from the skin surface in cattle. It is possible that the increase in peripheral blood TRP metabolites could also participate in the hypothermic effect of TRP.