Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass,and its badly preserved silage

In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.     

Effects of AI protocol and time interval from onset of estrus to AI on conception rate in Japanese Black cows

Two different artificial insemination (AI) protocols were investigated to determine the effect of the time interval from the onset of estrus (as detected by a pedometer) to the AI on the conception rate in Japanese Black cows. Seventy‐three cows were divided into two groups that received AI either after the induction of synchronized ovulation (synchronized AI group; n = 26) or at the time of observed estrus (control AI group; n = 47). Each group was further divided into two subgroups, which were artificially inseminated either 0–12 h (early AI group; n = 21) or 12–24 h (late AI group; n = 52) after the onset of estrus. There was no significant difference in the conception rate between the synchronized AI and control AI groups. The AI protocol and the detection of estrus using a threshold of counted steps (as measured by a pedometer) in this study could be effective for planned reproduction management without the observation of standing estrus in Japanese Black cows.     

Mean retention time of digesta in the different segments of the equine hindgut

Six Thoroughbred horses were used to determine the mean retention time (MRT) of digesta in the different segments of the hindgut. The horses were fed timothy hay or silage in equal amounts (1.6% bodyweight [BW]/day) every 3 h/day. Hay or silage labeled with the rare earth elements cerium, dysprosium, neodymium, ytterbium, lanthanum, samarium and praseodymium, were fed to the horses 36, 30, 24, 18, 12, 6 and 3 h before slaughter, respectively. Just after slaughter, digesta samples in the different segments were collected and determined for the contents of the rare earth elements. Using these data, a method to calculate the MRT in the different segments was proposed. There were no significant differences between diets in the MRT of each segment. The averaged MRT of hay and silage in the cecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon and small colon was 2.9, 3.1, 5.9, 1.0, 4.0, and 4.0 h, respectively. The dry matter weight of the digesta was related to the MRT in the right ventral colon (r = 0.94, P = 0.005), left ventral colon (r = 0.87, P = 0.03), left dorsal colon (r = 0.98, P < 0.001) and right dorsal colon (r = 0.95, P = 0.004), but it was not related to the MRT in the cecum and small colon.     

The functional effect of Gly209 and Ile213 substitutions on lysozyme activity of family 19 chitinase encoded by cyanophage Ma-LMM01

ORF69 in the cyanophage infecting Microcystis aeruginosa, Ma-LMM01, shows homology to the family 19 chitinases where the catalytic domain has structural similarity to lysozyme. Chitinases hydrolyze chitin, a β-1, 4-linked monopolymer of N-acetylglucosamine (GlcNAc); whereas lysozymes hydrolyzes peptidoglycan, alternating β-1, 4-linked copolymers of N-acetylmuramic acid (MurNAc) and GlcNAc. Using amino acid sequence comparison to ORF69, the putative sugar binding residues, Gln162 and Lys165, from the barley chitinase (the model enzyme for the family 19 chitinases) corresponding to subsites −4 and −3 were found to be replaced with Gly209 and Ile213, respectively, in ORF69. To analyze their contribution to substrate binding affinity, ORF69 was cloned into Escherichia coli; and two mutant proteins G209Q and I213K were prepared using site-directed mutagenesis. The wild-type gene product (gp69) showed both lysozyme and chitinase activities. In contrast, the I213K mutant showed a decrease (70%) in lysozyme activity and a significant increase (50%) in chitinase activity; whereas, the G209Q mutant almost completely abolished both enzyme activities. The data suggest the Ile213 residue is involved in recognizing the substrate MurNAc; and Gly209 has significant contribution in chitinase and lysozyme activities for the wild-type gp69.     

Allelopathic potential of itchgrass (Rottboellia exaltata L. f.) powder incorporated into soil

Itchgrass ( Rottboellia exaltata L. f.) is a widespread weed in northern Thailand. The farmers in this area have been using itchgrass as a mulching material in order to control other weeds in vegetable fields. Laboratory experiments were undertaken to investigate the phytotoxic activity of itchgrass powder incorporated into soil in order to evaluate the allelopathic activity in the field. The phytotoxic activity on the growth of radish seedlings ( Raphanus sativa L. var. radicula ), used as a test plant, was more pronounced in the root than in the shoot growth. The phytotoxic activity was found to be similar for the soils incorporated with the shoot or the root powder of itchgrass. The growth of the radish seedlings grown in sea sand and watered with soil water obtained from the soil previously incorporated with itchgrass powder showed a similar inhibition to those planted in the treated soil. The phytotoxic activity on the growth of the radish seedlings in the soil incorporated with the powder decreased over time. It is suggested that itchgrass releases phytotoxic compound(s) into soil water and the concentration of the active compound(s) in the soil water decreases over time.     

CLINICAL MAGNETIC RESONANCE VOLUMETRY OF THE HIPPOCAMPUS IN 58 EPILEPTIC DOGS*

Hippocampal atrophy, which is a component of hippocampal sclerosis and recognized commonly in human intractable epilepsy, is controversial in canine epilepsy. We examined the hippocampal volume in 58 epileptic dogs and 35 control dogs using magnetic resonance (MR) images, and calculated the relative hippocampal volume asymmetry of the right and left hippocampus. Subjectively, there were visible MR imaging abnormalities in seven of the 58 epileptic dogs (12%). The hippocampel volume asymmetric ratio of epileptic dogs (5.84±4.47%) was significantly greater than that of control dogs (1.62±0.88%). Using a cutoff threshold asymmetric ratio of 6% that is indicated in human epilepsy, 28 epileptic dogs (48%) were characterized as having unilateral hippocampus atrophy. The hippocampal volume asymmetry ratio cannot be used to detect bilateral atrophy. In conclusion, although less frequent than that observed in human epilepsy patients, hippocampal atrophy may occur in canine epilepsy.     

Influences of Northern Leaf Blight on corn silage fermentation quality,nutritive value and feed intake by sheep

This study examined the differences between non‐inoculated (control) corn and Northern Leaf Blight (NLB)‐damaged corn (inoculated corn); dry matter (DM) yield, silage fermentation quality, nutritive value and feed intake by sheep were compared. Leaf, stem and grain dry weights and gross yield of inoculated corn were significantly (P < 0.05) decreased compared with control corn. The contents of water‐soluble carbohydrate and nitrogen‐free extract (NFE) were decreased in inoculated corn compared with control corn. Silage made from both inoculated and control corn showed good fermentation quality. The digestibility of DM, organic matter, ether extract, NFE, and energy of silage made from inoculated corn were significantly (P < 0.05) lower, and contents of total digestible nutrients (TDN) and digestible energy (DE) were also significantly (P < 0.05) lower compared with silage made from the control corn. DM intake showed no significant discrepancy between the two types of silage. TDN and DE intakes from inoculated silage were significantly (P < 0.05) lower compared with control silage. From the above results it was shown that NLB caused a decrease in DM yield and NFE content in corn and a decrease in the nutritive value and feed intake of silage.     

日本奶农生产经营的改善及其对我国奶业发展的启示

日本奶牛的饲养主要依赖于国外饲料的进口,国际饲料价格的上涨使得日本奶业陷入了危机,奶农的生产经营状况直接关系到奶业产业的发展。本研究基于日本社团法人中央畜产会实施的全国农户生产经营指导项目中所调查的294个奶农农户的生产经营相关数据,采用因子分析、多元线性回归分析、聚类分析方法,对影响奶农经营收入的因素进行了分析。结果表明,奶牛饲养的收益、奶牛饲养的劳动效率、饲养规模这三个因素对奶农的生产影响最大;阐明了奶农生产中存在的问题;提出了奶农生产经营的改善应该着重于适度扩大生产规模;合理进行投资,并且均衡大型机器设备和小农器具投资的比例以改善奶农饲养效率以及饲料生产效率,提高奶农的收入。最后,借鉴日本奶业生产经营中存在的问题,总结了对我国奶业发展的启示。     

Squamous cell carcinoma in a digit of the hind limb with systemic metastasis in a 17-year-old female koala

We encountered a case of cutaneous squamous cell carcinoma (SCC) in a 17-year-old female koala at a zoo. A fragile, papillary, elevated mass was found on the third digit of the right hind limb. SCC was identified histopathologically: squamous cell-like polygonal tumor cells showed a nest-like growth pattern with epidermal down growth, central keratinization and necrotic foci, and invaded dermal connective tissues. Metastatic lesions were observed in various organs, including the lung and axillary lymph node: in the lung, multiple metastatic foci similar to the primary lesion, and in the axillary lymph node, individual polygonal tumor cells infiltrated the sinusoids. Immunohistochemistry revealed that the tumor cells were positive for proliferating cell nuclear antigen, which exhibited 32–33% of labeling indices in the tumor cells. To our knowledge, this is the first report of a case of SCC in a digit of a koala.