腹腔注射N-甲基-D-天冬氨酸对母鼠发情率、早期胚胎及胚子数的影响

为研究腹腔注射N-甲基-D-天冬氨酸(NMDA)对母鼠发情率和早期胚胎及胚子数的影响,试验选取40只昆明小鼠[(26±1.37)g],随机分为对照组和试验组(每组各20只),对照组腹腔注射0.2mL生理盐水,试验组腹腔注射0.2mLNMDA溶液,采用棉签蘸取母鼠阴道黏液,涂片后用苏木精(HE)染色,判定母鼠发情情况。随后在发情母鼠与公鼠合笼后第3天,每组随机取10只颈椎脱臼法处死小鼠,冲胚后在体视镜下捡胚观察胚胎个数;第9天处死剩余小鼠,取出子宫统计胚子数。试验结果显示,腹腔注射NMDA试验组母鼠发情率、胚胎数和胚子数分别比对照组提高28.57%、18.18%和12.94%。该试验表明腹腔注射NMDA可促进母鼠的发情,提高早期胚胎和胚子数,为NMDA在动物繁殖中的应用提供了理论基础。     

Complement of integrated fertilizer management and integrated pest management concepts to ameliorate faba bean growth and yield

Twenty-one microbial preparations recommended for controlling pathogenic fungal strains causing root rot and wilt diseases of faba bean were investigated for antibiosis against several symbiotic and associative diazotrophs adopting a modified agar-plate-inhibition-zone assay. Rhizobium and Bradyrhizobium exhibited a somewhat similar susceptibility to biocontrol agents while associative diazotrophs showed variable responses. Azotobacter, compared to others, was severely inhibited by such bio-candidates. The members of the biofertilizer formulation ‘Biofertan’ did bear mixed cultivation with the majority of biocontrol agents. Among those, Bacillus subtilis was deemed the pioneer. In pot experiments, almost all the antagonists significantly restricted the severity of root rot and wilt diseases besides modifying faba bean seedling stand and improving plant development. This was very obvious with shoot biomass increases of >?50%. Moreover, the bioagents successfully recovered the legume establishment, seriously injured due to pathogenic fungal infection. Simultaneous inoculation with Rhizobium and biocontrol agents provided more growth stimulation compared to either when introduced individually. Field-grown faba beans were inoculated with the diazotroph and representatives of biocontrol strains by two different methods, seed coating and over-head soil. Growth parameters determined were the highest when the legume plant was seed-coated by Rhizobium simultaneously over-head soil inoculated with the bacterial bioagents; this was reported with Bacillus subtilis and Pseudomonas aerugenosa. The significance of combined application of biofertilizer and bioagent to ensure cheap, clean and safe farm products is discussed.     

Pregnancy-related diabetes mellitus in two dogs

CASE SUMMARIES: Two cases of diabetes mellitus occurring in bitches in association with pregnancy are reported. In the first case, a bitch with suspected acromegaly developed diabetes mellitus within 2 weeks of the due date. Despite insulin therapy, euglycaemia was not achieved. Tw o live, small pups were delivered by elective Caesarean section but died within 2 days. Signs consistent with acromegaly resolved but diabetes mellitus was permanent in the bitch. In the second case, diabetic ketosis with severe gastrointestinal disease was diagnosed 2 days after Caesarean section was performed due to dystocia. The pups delivered all died within 5 days. The bitch recovered fully from diabetes mellitus within 2 weeks and has remained euglycaemic without insulin for a period of at least 18 months.

CLINICAL RELEVANCE: These two cases demonstrate that diabetes mellitus can occur in association with pregnancy in dogs, that diabetic ketosis can occur during transient diabetes mellitus in dogs, and suggest that acromegaly may occur during pregnancy-related dioestrus in dogs. The scarcity of previous reports of this nature, however, suggests that such cases are unusual.

Lack of prompt resolution of hyperglycaemia may result in secondary diabetes mellitus becoming permanent. Management should focus on immediate insulin therapy or ovariohysterectomy to minimise this risk. Even mild hyperglycaemia should not be ignored during pregnancy. The insulin antagonistic effects of pregnancy, stressful illness, surgery and dystocia can be enough to result in diabetic ketosis in the absence of permanent insulin deficiency. Maternal hyperglycaemia may contribute to adverse fetal outcomes in dogs but further study is required regarding the nature of the risk.     

Detection of Water Buffalo Sex Chromosomes in Spermatozoa by Fluorescence in situ Hybridization

In order to identify X‐ and Y‐bearing spermatozoa in water buffalo by fluorescence in situ hybridization (FISH), some available probes of closely related species were examined. An X‐ and Y‐specific probe set, made from flow sorted yak chromosomes, labelled in somatic metaphases of water buffalo the whole X and Y, respectively, except their centromere regions. A cattle Y‐chromosome repeat sequence (BC1.2) showed strong signal on the telomere region of the buffalo Y‐chromosome, demonstrating the evolutionary conservation of this locus in water buffalo. In hybridization experiments with spermatozoa from five buffaloes, the yak X‐Y paint set demonstrated clear signals in more than 92% (46.8% X and 45.8% Y) of the cells. Using the cattle Y‐chromosome specific BC1.2 probe, clear hybridization signal was detected in more than 48% of the cells. Statistical analysis showed that there was no significant difference between bulls or from the expected 50 : 50 ratio of X‐ and Y‐bearing cells. The probes presented here are reliable to assess separation of X‐ and Y‐bearing spermatozoa.     

Successful Vitrification of In vivo Embryos Collected from Superovulated Japanese Black Cattle (Wagyu)

The aim of this study was to determine whether vitrification is an effective method when used for Japanese Black Cattle (Wagyu) in vivo‐derived embryos, collected following a superovulation treatment and embryo transfer (MOET) programme. In vivo‐derived morula and blastocysts collected on day 7 after artificial insemination, were vitrified using a modified droplet vitrification (MDV) procedure and subsequently warmed for transfer (ET) into synchronized recipients. Fresh embryos, and embryos cryopreserved using a standardized slow freezing procedure (direct thaw/direct transfer, DT) served as ET controls. Two different follicle‐stimulating hormone (FSH) sources, Folltropin^® Canada (FSH BAH, 24 donors) and a brand prepared by the Chinese Academy of Science (FSH CAS, 16 donors), were compared in a series of superovulation outcomes following well‐established FSH administration protocols. Following data analysis, the total number of ovulations recorded at the time of embryo flushing (10.5 vs 8.5; p = 0.28) and the total number of transferable embryos (6.2 vs 5.1; p = 0.52) were similar between the two FSH sources. ET for MDV (39.7%, n = 78), DT (35.2%, n = 71) and fresh controls (47.1%, n = 34) resulted in similar pregnancy rates (p > 0.05). When MDV was used, a higher pregnancy rate (42.6%) resulted from the transfer of vitrified morulae, when compared to the DT counterparts (24.3%), (p = 0.05). Transfer of vitrified morulae resulted also in higher pregnancy rate, when compared to the transfer of vitrified blastocysts (42.6% vs. 29.4%; p < 0.05). Transfer of DT blastocysts resulted in higher pregnancy rate than morulae, similarly cryopreserved (47.1% vs. 24.3%, p < 0.05). In conclusion, MDV is an effective alternative methodology for cryopreservation of in vivo‐derived embryos. This study gives also indication that, compared to vitrified blastocysts, MDV of morula stage embryos results in higher pregnancy rates following warming and transfer into synchronized recipients.     

茵山莲水提物对四氯化碳诱导小鼠急性肝损伤的保护作用

为探讨茵山莲水提物对四氯化碳(CCl₄)诱导的小鼠急性肝损伤的保护作用及可能机制,以对后续茵山莲的深入研究及临床转化提供理论指导。将60只ICR小鼠随机分为6组,每组10只,分别为空白对照组、模型对照组、联苯双酯对照组、茵山莲低(1.17 mg·g^-1)、中(2.34 mg·g^-1)、高(4.68 mg·g^-1)剂量组,按不同剂量药物每天灌胃1次,给药7 d。在末次给药1 h后,除空白对照组以外,其余各组小鼠腹腔注射0.2%四氯化碳构建急性肝损伤模型。12 h后小鼠眼球采血并分离血清,收集肝组织。采用生化分析检测血清总蛋白(TP)和谷胱甘肽S转移酶(GST)活性,肝组织中GST、琥珀酸脱氢酶(SDH)、过氧化氢酶(CAT)和总超氧化物歧化酶(T-SOD)活性,利用HE染色和Masson染色观察肝组织病理及肝纤维化情况,采用蛋白免疫印迹法和免疫组织化学法检测肝组织白介素1beta (IL-1β)、肿瘤坏死因子alpha (TNF-α)、血管生成因子A (VEGFA),金属基质蛋白酶9(MMP9)及转录因子NF-κBp65、蛋白激酶IKK、p38MAPK等蛋白表达及定位情况。结果表明,与模型对照组相比,2.34 mg·g^-1茵山莲水提物能显著降低小鼠血清GST水平,显著升高肝内CAT水平,各组间T-SOD无显著差异,但2.34 mg·g^-1茵山莲水提物组小鼠与空白对照组、模型对照组在肝GST、SDH和血清TP水平上无显著差异。2.34 mg·g^-1茵山莲水提物能缓解CCl₄诱导的小鼠肝病理损伤和纤维化病变,抑制肝IL-1β、TNF-α、VEGFA、NF-κBp65和IKK的表达。提示茵山莲水提物可能通过NF-κB和p38MAPK通路抑制肝氧化应激和炎症水平,从而缓解小鼠急性肝损伤。     

Agreement among three different equine venipuncture sites with regard to measurement of packed cell volume and total solids

The primary objective of this study was to determine the agreement between the packed cell volume (PCV) and total solids (TS) values in blood collected from the jugular vein (JV) in comparison with the cephalic vein (CV) and the transverse facial venous sinus (TFVS) in healthy adult horses. A total of 72 samples were taken from 24 horses. We found high agreement that reflects no clinically relevant differences between the PCV and TS values obtained from the CV or the TFVS in comparison with the JV in standing healthy adult horses.