Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Presentation,clinical pathological and post‐mortem findings in three related Scottish terriers with ligneous membranitis

Ligneous conjunctivitis and gingivitis were diagnosed in three related Scottish terrier dogs presented for investigation of severe conjunctivitis and respiratory signs. Hypoplasminogenaemia was confirmed in one of the three affected dogs. Supportive treatment was not effective, and the dogs died or were euthanased because of the disease. Post‐mortem analysis of two of the dogs revealed multiple abnormalities including severe proliferative fibrinous lesions affecting the conjunctiva, gingiva, trachea, larynx and epicardium and multiple fibrous adhesions throughout the thoracic and abdominal cavities. One dog had internal hydrocephalus and lacked a cerebellar vermis. Ligneous membranitis was confirmed on histopathology. This is a rare condition in dogs but an important differential diagnosis for severe conjunctivitis and gingivitis.     

Surveillance of dairy production holdings supplying raw milk to the farmhouse cheese sector for Escherichia coli O157, O26 and O111

Clinically healthy domestic animals can harbour Escherichia coli O157 and other verocytotoxigenic E. coli (VTEC) strains in their faeces. Milk filters can be used to microbiologically monitor direct milk secretion and environmental contamination for these pathogens. The aim of this study was to establish baseline data on the prevalence and characteristics of VTEC organisms in lactating animals (bovine, ovine and caprine) supplying milk to the farmhouse cheese sector, with particular emphasis on serogroups O157, O111 and O26. Fifty-six bovine, 13 caprine and 5 ovine herds/flocks, the majority of which supplying milk for farmhouse cheese production, were surveyed from May 2004 to July 2005. Milk filters were analysed by immunomagnetic separation followed by PCR, on a serogroup-specific basis for E. coli O157, O26 and O111. Positive isolates were examined using a multiplex PCR protocol, for their potential to produce verocytotoxins (vt1/vt2), the haemolysin-encoding gene (hlyA) and the gene encoding attaching and effacement (eae). Five verocytotoxigenic and 22 non-virulent E. coli O157 isolates were detected. Seventeen E. coli O26 isolates were also detected, four of which were verocytotoxigenic, seven isolates contained the eae gene only and six isolates were devoid of any of the virulence factors. The VTEC O157 and O26 isolates contained the hlyA and eae genes along with the verocytotoxin genes. No E. coli O111 isolates were detected. Some of the herds were positive on more than one occasion and multiple E. coli serogroups were isolated from the same milk filter sample. Although all food products tested were VTEC negative, routine surveillance for such pathogens in raw milk/raw milk products is of public health importance. Herd-level surveillance along with subsequent risk management action may be a cost-effective component of risk reduction strategies for food production, drinking water supplies and the protection of public health.     

不同质地土壤玉米秸秆还田配施腐熟剂效应的研究

为研究鉴选不同质地土壤秸秆还田适宜的秸秆腐熟剂,于2017—2018年在西辽河平原灌区中壤土和砂壤土秸秆还田配施人元腐熟剂和中农绿康腐熟剂,以秸秆还田不施腐熟剂为对照,共设置6个处理,测定玉米秸秆腐解率、玉米产量、玉米根系特性和土壤化学性状,研究不同秸秆腐熟剂对玉米产量及土壤特性的影响。结果表明:不同质地土壤2种腐熟剂秸秆腐解率间无显著差异;中壤土2年各处理玉米产量及根系特性、土壤化学性状均表现为人元腐熟剂中农绿康腐熟剂秸秆还田不施腐熟剂,且中农绿康腐熟剂和人元腐熟剂显著高于秸秆还田不施腐熟剂,而2种腐熟剂处理之间无显著差异;砂壤土2年各处理玉米产量及根系特性、土壤化学性状均表现为中农绿康腐熟剂人元腐熟剂秸秆还田不施腐熟剂,且中农绿康腐熟剂显著高于人元腐熟剂和秸秆还田不施腐熟剂。2年不同质地土壤中配施腐熟剂在根系特性、土壤化学性状方面表现为砂壤土配施中农绿康腐熟剂效果优于人元腐熟剂,而中壤土配施2种腐熟剂效果无显著性差异。     

“珍珠”的工艺配方研究

通过正交试验,对"珍珠"的工艺配方进行了研究,结果表明,胶液最佳配方为海藻酸钠∶明胶∶琼脂为3∶1∶2,奶油含量5%,蔗糖含量10%。氯化钙浓度3.0%,固化时间10min。得到的"珍珠"颗粒形状圆润,口感滑嫩,3个月内不发生溶解。     

长爪沙鼠颌下腺免疫球蛋白A的定位分布

采用免疫组织化学方法对不同日龄的长爪沙鼠颌下腺IgA的定位分布进行了研究。结果表明，长爪沙鼠的颌下腺由导管部和分泌部构成，分泌部主要由浆液腺构成，导管部包括闰管、纹管、颗粒曲管和小叶间导管等。DAB显色结果表明，IgA阳性细胞主要分布于浆液性腺泡、闰管、纹管、颗粒曲管和小叶间导管，并可分布于腺泡和腺管间结缔组织，IgA阳性产物的分布具有不均一性，无明显随年龄变化的规律性。阳性产物分布于胞质中，胞核呈阴性，对照组阴性。提示从浆细胞产生或循环而来的IgA先经结缔组织进入颌下腺组织，进而定位分布于浆液腺泡和各级导管，导管部有较多的IgA分布。