One year investigation of the prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido

We investigated the seasonal prevalence and diversity of clostridial spores in raw milk from the Tokachi area of Hokkaido. Samples of raw milk were collected quarterly from May 2013 through February 2014. The mean clostridial spore count for the raw milk from 336 milk tankers was 27.6 CFU/100 ml. The clostridial species isolated most frequently from raw milk samples was Clostridium tyrobutyricum. The dominant species was C. tyrobutyricum regardless of the season. The percentage of samples with low spore counts (<10 CFU/100 ml) was highest (60.9%) during winter (February) and lowest (34.5%) in autumn (November). In comparison, the percentage of samples with high spore counts (>100 CFU/100 ml) was highest (5.7%) in autumn (November) and lowest (1.1%) during spring (May).     

Developmental ability of oocytes retrieved from Meishan neonatal ovarian tissue grafted into nude mice

Ovarian xenografting makes it possible to obtain oocytes with fertilization ability from immature pigs of Western breeds. In this study, we applied these methods to the Meishan, an indigenous Chinese pig breed, and investigated the developmental competence of oocytes grown in their neonatal tissue after grafting into nude mice. First, mice harboring neonatal ovarian tissue were infused with follicle stimulating hormone (FSH) (62.5 U/ml) for 13 days starting at 10, 30, and 60 days after vaginal opening (D10‐, D30‐, and D60‐FSH groups, respectively). Development of antral follicles and their oocytes was most enhanced in the D60‐FSH group. For the next step, we examined the in vitro maturation ability of the oocytes recovered from host mice after infusion with FSH at a dose of 62.5 U/ml or 125 U/ml (FSH‐62.5 or ‐125 group) for 13 days starting at 60 days after vaginal opening. Many more oocytes with maturation ability were obtained from the FSH‐125 group. The FSH‐125 mature oocytes were fertilized in vitro, as shown by formation of male and female pronuclei, but did not reach the blastocyst stage. These results indicate that Meishan neonatal ovaries are able to produce oocytes with fertilization ability after being grafted into nude mice.     

Presence of chlorogenic acid during in vitro maturation protects porcine oocytes from the negative effects of heat stress

Chlorogenic acid (CGA) is known to protect oocytes from oxidative stress. Here we investigated the effects of CGA on porcine oocyte maturation under heat stress and subsequent embryonic development after parthenogenetic activation. For in vitro maturation (IVM) at 41.0°C (hyperthermic condition), supplementation of the maturation medium with 50 μM CGA significantly improved the percentage of matured oocytes and reduced the rate of apoptosis relative to oocytes matured without CGA (p < .05). CGA treatment of oocytes during IVM under hyperthermia tended to increase (p < .1) percentage of blastocyst formation after parthenogenesis and significantly increased (p < .05) the total cell number per blastocyst relative to oocytes matured without CGA. For IVM at 38.5°C (isothermic condition), CGA significantly improved the rate of blastocyst development compared with oocytes matured without CGA (p < .05), but did not affect oocyte maturation, apoptosis rate or the number of cells per embryo. Omission of all antioxidants from the IVM medium significantly reduced the rate of oocyte maturation, but the rate was restored upon addition of CGA. These results demonstrate that CGA is a potent antioxidant that protects porcine oocytes from the negative effects of heat stress, thus reducing the frequency of apoptosis and improving the quality of embryos.     

Provision of beta‐glucan prebiotics (cellooligosaccharides and kraft pulp) to calves from pre‐ to post‐weaning period on pasture

We conducted two feeding experiments to evaluate the effects of supplementation with either cellooligosaccharide or kraft pulp on growth performance in grazing beef calves (Japanese Black) from 4 weeks pre‐weaning to 12 to 16 weeks post‐weaning. In Experiment 1 (20‐week duration), nine calves (2.9‐month‐old females) were assigned to either a control group (CON) or an experimental group (CEL) fed cellooligosaccharide at a rate of 10 g/day mixed with concentrate. Average daily weight gain tended to be greater in CEL than in CON, especially after 1 month of weaning. In Experiment 2 (16‐week duration), 10 calves (2.0‐month‐old females) were assigned to either a control group or an experimental group (KRA) fed kraft pulp at a rate of 10% replacement of total digestible nutrients with concentrate. The proportion of fibrolytic bacteria increased and that of methanogenic Archaea decreased in the rumen microbial community composition of KRA calves in Experiment 2, whereas the decrease in Fibrobacter and Archaea was observed in CEL calves at first 4 weeks in Experiment 1. We conclude that beta‐glucan prebiotic supplementation to grazing calves at pre‐weaning would affect rumen microbial composition and modified rumen fermentation characteristics, leading to a better rumen environment via different means.     

Harpinpsta from Pseudomonas syringae pv. tabaci Is Defective and Deficient in Its Expression and HR-inducing Activity

To elucidate the role of harpins produced by Pseudomonas syringae, the corresponding hrpZ gene was isolated from P. s. pv. tabaci. The sequence information revealed that this gene carries a serious mutation with 326 bp lacking in the central region and potentially encodes only 140 N-terminal amino acids because of a frame shift. The investigation of biological properties using recombinant harpin indicated harpin_psta was incapable of inducing HR in both host and nonhost plants. Based on an immunoblot analysis to detect harpin from P. s. pathovars in hrp-inducing medium, the truncated harpin_psta was neither expressed nor secreted into the culture medium. These results suggest that harpin is not the sole determinant of the host-parasite specificity in P. s. pv. tabaci. Received 10 August 2000/ Accepted in revised form 21 December 2000     

Analysis of factors affecting milking claw vacuum levels using a simulated milking device

Bovine mastitis is typically caused by microbial infection of the udder, but the factors responsible for this condition are varied. One potential cause is the milking system, and although previous studies have investigated various methods for inspecting these devices, most have not assessed methods for evaluating the milking units. With this in mind, we analyzed the factors that affect the vacuum inside the milking claw by using a simulated milking device and by measuring milking claw vacuum when adjusting the flow rate in five stages. The factors analyzed in each milking system were the vacuum pressure settings (high and low line system) , milk tube length (200‐328 cm), aperture diameter (14‐22.2 mm), constricted aperture diameter (12 mm), tubing configurations, lift formation (0‐80 cm), claw type (bottom and top flow) and use or non‐use of a milk sampler. The study findings demonstrated that all of these variables had a significant impact on claw vacuum and suggest that a diagnostic method using a simulated milking device should be considered when inspecting modern milking systems.     

Randomized clinical trial to evaluate the effectiveness of enrofloxacin as a second‐line antibiotic for treatment of acute Escherichia coli mastitis

The objective of the present study was to evaluate the effectiveness of enrofloxacin (ERFX) as a second‐line antibiotic for treatment of acute Escherichia coli (E. coli) mastitis. Forty‐two cows with naturally occurring acute E. coli mastitis were enrolled. On the first day of treatment (day 0), empirically selected antibiotics (oxytetracycline: n = 32, kanamycin: n = 10) were administered. Although systemic signs improved in 10 cows (first‐line group), the signs remained unchanged or worsened in 32 cows on day 1, including two cows that were found dead. The 30 surviving cows were randomly assigned to second‐line groups constituting an ERFX group (n = 19) or a control group (n = 11) that was treated with other antibiotics. Response to each treatment was evaluated by measuring clinical signs from day 0 to day 3, subsequent quarter milk recovery, and the 60‐day survival rate. Appetite on day 3 was significantly better in the ERFX group compared to the control group. No significant differences were observed in the 60‐day survival rate or the subsequent milk recovery between the ERFX group and the control group. Thus, the use of ERFX as a second‐line antibiotic for the treatment of acute E. coli mastitis could induce a rapid appetite recovery.     

Antimicrobial activity of heartwood components of sugi (Cryptomeria japonica) against several fungi and bacteria

Methanol extract of sawdust of sugi (Cryptomeria japonica) heartwood was fractionated with toluene and n-hexane to give solvent-soluble and solvent-insoluble fractions. The n-hexane-soluble fraction showed the most inhibition activity among the fractions against phytopathogenic microorganisms, namely Fusarium oxysporum, Phytophthora capsici, Pythium splendens, and Ralstonia solanacearum. Sandaracopimarinol and ferruginol, isolated from the n-hexane-soluble fraction, showed moderate antifungal activity against the three fungi and strong antibacterial activity against R. solanacearum. The content of sandaracopimarinol (7.07 g/kg based on the dried sawdust) in the heartwood was about twice that of ferruginol. Sandaracopimarinol and ferruginol strongly inhibited the growth of Gram-positive bacteria but did not show inhibitory action against Gram-negative bacteria except for R. solanacearum. The antibacterial effect of sandaracopimarinol was first found in the present study and was stronger than that of ferruginol.     

Forest road design with soil sediment evaluation using a high-resolution DEM

In this study, we combined a program to optimize forest road alignments with a method for prediction of surface erosion and related sediment delivered to streams. Combining the forest road design program with the high-resolution digital elevation model made it possible to estimate soil sediment based on a standard methodology, because a relatively accurate road prism could be generated. The combined program properly places and spaces drainage structures based on the Washington State Forest practice board manual. We applied the program to a part of Capitol State Forest in Washington State and optimized horizontal and vertical alignments while estimating movement of soil sediment from roads to streams. We discussed the effects of road surface materials, near-stream culvert distance to stream, and out-sloped forest road template on total road costs and soil sediment delivered to streams. Using lower quality rock surfacing reduced total costs, but the amount of soil sediment from lower quality rock surfacing was 1.5 times more than that on a higher quality rock traction surface. Therefore, lower quality rock surfacing should not be used near the stream. The placement of near-stream culverts 15 m upstream and out-sloped road template were examined using a traction surface on 15-m sections upstream and lower quality rock surfacing on other sections in order to reduce soil sediments to the stream and total costs. As a result, placing near-stream culverts 15 m upstream and using an out-sloped road template significantly reduced total road cost and soil sediment. Finally, we optimized horizontal and vertical alignments while limiting soil sediment to a specified maximum level. The model successfully optimized forest road alignments, which reduced total road cost as well as soil sediment.