Detection of DNA restriction fragment polymorphisms in Streptococcus equi

Large-restriction-fragment (LRF) polymorphisms in Streptococcus equi (S equi subspecies equi) were studied by pulsed-field gel electrophoresis. Five or six chromosomal fragments of between 194 and 915 kb were separated by digestion with the restriction endonuclease Notl. All 20 isolates of S equi, including 12 from independent Japanese outbreaks, four from independent American outbreaks, two from a single Irish outbreak, us vaccine strain F43, and type strain NCTC 9682 were successfully typed. Seven distinctive, reproducible and stable types were identified. The 12 Japanese isolates collected between 1992 and 1998 were of LRF type II suggesting that they were derived from the same source. The remaining eight isolates were of six types. The results indicate that LRF typing should be a useful technique for investigating the source and transmission of S equi.     

Relationship between the first ovulation within three weeks postpartum and subsequent ovarian cycles and fertility in high producing dairy cows

The aim of this study was to investigate the relationship between the first ovulation within 3 weeks postpartum and subsequent ovarian cycles and fertility in high producing dairy cattle in Hokkaido, Japan. In Experiment 1, 110 cows (44 primiparous and 66 multiparous) were used to determine the effects of the first ovulation within 3 weeks postpartum on subsequent ovarian cycles. Milk samples were collected twice weekly from 7 to 100 days postpartum. The first ovulation was identified by an increase in milk progesterone (P4) to more than 1 ng/ml within 3 weeks postpartum. The numbers of cows showing ovulation and anovulation within 3 weeks postpartum were 31 (70.5%) and 13 (29.5%) in the primiparous cows and 35 (53.0%) and 31 (47.0%) in the multiparous cows, respectively. The patterns of ovarian resumption after calving were classified into two types (normal ovarian cycles and abnormal ovarian cycles) on the basis of milk P4 concentrations. Initiation of normal ovarian function in cows ovulated within 3 weeks postpartum occurred earlier than in anovulated cows regardless of the number of calvings (primiparous, 27.8 days vs. 44.4 days; multiparous, 30.6 days vs. 55.7 days; P<0.01). Out of the multiparous cows that ovulated within 3 weeks postpartum, initiation of normal ovarian function followed by a normal luteal phase was earlier than when it was followed by an abnormal luteal phase (25.5 days vs. 40.4 days; P<0.05). Milk P4 concentrations after the first ovulation were lower than those after the second ovulation in both the primiparous and multiparous cows (P<0.05). In Experiment 2, 22 multiparous cows were used to determine the effects of the first ovulation within 3 weeks postpartum on subsequent fertility. Blood samples were collected once a week from 0 to 3 weeks postpartum. The interval from parturition to first service in ovulated cows was shorter than in anovulated cows (68.4 days vs. 94.8 days; P<0.05). The conception rate by 100 days after calving tended to be higher in ovulated cows than in anovulated cows (50.0% vs. 16.7%, P=0.09). In conclusion, our data strongly suggests that ovulation within 3 weeks postpartum is a crucial phenomenon for subsequent resumption of ovarian function and conception, and thus it can be used as an index of subsequent reproductive performance.     

Movements of dams milked for fermented horse milk production in Mongolia

Airag, (Fermented horse milk) is a traditional milk product in Mongolia. Herders separate foals from their dams and tie them at a milking site during the daytime to produce airag. To evaluate the effects of horse management on the movement of dams, we tracked three dams in a herd in camp 1 during summer and camp 2 during autumn of 2013 and analyzed their movements during the milking (daytime) and non‐milking (nighttime) periods in an area famous for its high‐quality airag. Dams were gathered every 1.7 ± 0.0 h between 07.46 and 15.47 hours at the milking sites and milked 4.6 ± 0.2 times/day during the study period (86 days). Daily cumulative and maximum linear distances from the milking sites were longer (P < 0.01) during the non‐milking period than during the milking period. Daily home ranges were 91 and 26 times greater during the non‐milking period (P < 0.001) in camps 1 and 2, respectively. The greater range during the non‐milking period would reflect the spatial distributions of water, salt and forage. The dams initially used similar areas and gradually shifted their daily home ranges after several days. This shift suggests that the dams grazed farther afield as forage availability declined around the milking site. For better airag production and sustainable pasture use, our results provide insights useful for evaluating the effects of milking management on vegetation and soil in those pastures, for selecting the appropriate milking times and frequency, and for choosing the right timing to shift milking sites.     

Effect of dietary sorbitol on the performance of broilers

1. The effect of dietary sorbitol on the performance of broilers was investigated. Male and female broilers were given a control diet or a sorbitol diet (100 g/kg diet) ad libitum from 29 to 57 d of age. 2. Body weight gain, food intake, food efficiency and the ME values of the diets were not significantly different. Absolute and relative weights of abdominal fat were significantly reduced in birds given the sorbitol diet. 3. Compared to the control diet, the diet containing sorbitol lowered the serum glucose, total cholesterol and very low density lipoprotein concentrations.     

Epigenetic assessment of environmental chemicals detected in maternal peripheral and cord blood samples

Epigenetic alteration is an emerging paradigm underlying the long-term effects of chemicals on gene functions. Various chemicals, including organophosphate insecticides and heavy metals, have been detected in the human fetal environment. Epigenetics by DNA methylation and histone modifications, through dynamic chromatin remodeling, is a mechanism for genome stability and gene functions. To investigate whether such environmental chemicals may cause epigenetic alterations, we studied the effects of selected chemicals on morphological changes in heterochromatin and DNA methylation status in mouse ES cells (ESCs). Twenty-five chemicals, including organophosphate insecticides, heavy metals and their metabolites, were assessed for their effect on the epigenetic status of mouse ESCs by monitoring heterochromatin stained with 4￠,6-diamino-2-phenylindole (DAPI). The cells were surveyed after 48 or 96 h of exposure to the chemicals at the serum concentrations of cord blood. The candidates for epigenetic mutagens were examined for the effect on DNA methylation at genic regions. Of the 25 chemicals, five chemicals (diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se) and octachlorodipropyl ether (S-421)) caused alterations in nuclear staining, suggesting that they affected heterochromatin conditions. Hg and Se caused aberrant DNA methylation at gene loci. Furthermore, DEP at 0.1 ppb caused irreversible heterochromatin changes in ESCs, and DEP-, Hg- and S-421-exposed cells also exhibited impaired formation of the embryoid body (EB), which is an in vitro model for early embryos. We established a system for assessment of epigenetic mutagens. We identified environmental chemicals that could have effects on the human fetus epigenetic status.     

Development of a new selective medium for isolation of methicillin-resistant Staphylococcus aureus

A new selective medium containing cephem antibiotics was developed for isolation of methicillin-resistant Staphylococcus aureus (MRSA). MRSA colonies on a medium containing ceftazidime (CAZ) were most easily identifiable and a medium containing cefoperazone (CPZ) was superior in suppressing the growth of other bacteria. With the medium containing a couple of CAZ and CPZ, MRSA and methicillin-resistant coagulase-negative staphylococci (MRCNS) were detected from 2 and 1 of 15 chicken meat samples respectively. The MRSA and MRCNS recovery test showed that the medium was effective for MRSA isolation, suppressing the growth of other bacteria efficiently. These results suggested that the medium containing a couple of CAZ and CPZ was useful for MRSA detection from foods and animals.     

Peripartum back fat thickness of multiparous Holstein-Friesian cows with displacement of the abomasum or ketosis

To establish a method to predict postpartum diseases using prepartum back fat thickness (BFT), the peripartum BFTs of 54 healthy multiparous cows before calving, which were diagnosed with postpartum displacement of the abomasum (DA), clinical ketosis or subclinical ketosis were compared with those of healthy cows from 8 weeks before the expected calving date to 8 weeks after calving. The peripartum BFTs of the cows with DA or clinical ketosis were significantly higher than those of healthy cows. The peripartum BFTs of the cows with subclinical ketosis were not significantly higher than those of the healthy cows.     

Characterization and classification of Actinobacillus (Haemophilus) pleuropneumoniae plasmids.

Actinobacillus (Haemophilus) pleuropneumoniae plasmids were characterized and classified. They were isolated from A pleuropneumoniae strains different in serotype, year isolated, or location from which isolated. Six of 8 plasmids encoded streptomycin (Sm) and sulfonamide (Su) resistance (SmSu). One of the other plasmids, pVM105, encoded ampicillin (Ap) resistance and another, pHM0, encoded no drug resistance. All SmSu plasmids were transferred to Escherichia coli strains by transformation. Among them, pABO and pMS260 were 8.1 kb and incompatible with each other; they were stable in E coli. The other SmSu plasmids, pHM1, pVM104, pVM106, and pKD25, were 4.3 kb and did not replicate stably in E coli. The former SmSu plasmids were mobilized in E coli strains by a plasmid RP4, which belonged to incompatibility (Inc) group P, but the latter plasmids were not. Further, each 8.1-kb SmSu plasmid and each 4.3-kb plasmid had the same respective restriction pattern. These results indicated that there were at least 2 types of SmSu plasmids in A pleuropneumoniae. The 2 types were classified in 2 groups: H1(pMS260 and pABO) and H2(pHM1, pVM104, pVM106, and pKD25). The H1 and H2 plasmids belonged to different Inc groups, and H2 plasmids belonged to a different Inc group from that of pHMO and pVM105.     

Localization of cytokines in tendinocytes of the superficial digital flexor tendon in the horse

Although inflammatory activation of cytokines have been analyzed in various tissues, there have only been a few and as-yet-inconclusive studies on cytokines in equine tendons. In this study, the localizations of 4 cytokines (IL-1alpha, IL-1beta, TNFalpha and IFNgamma) in tendinocytes of the equine superficial digital flexor tendon (SDFT) were analyzed by the use of an immunohistochemical method. In inflamed tendons positive staining for all 4 cytokines antibodies were detected in endotedinieum cells and vascular epithelial cells. In contrast, negative or trace immunoreactions were obtained in many tendinocytes in the normal tendon. The variation in cellular immune responses depending on the kind of cytokine may reflect the physiological/pathological condition of the SDFT.