Detection of papillomavirus-DNA in mesenchymal tumour cells and not in the hyperplastic epithelium of feline sarcoids

Abstract We examined 12 formalin-fixed paraffin-embedded feline skin tumours which had the histopathological features of fibropapillomas for the presence of papillomavirus (PV) DNA using touchdown polymerase chain recation (PCR), DNA sequencing and nonradioactive in situ hybridization. Nine of the tumours contained a 102-bp PCR product demonstrated using consensus PV primers that amplify a portion of the L1 gene. The nucleotide sequences are closely related, but not identical to that of ovine PV type 2, rabbit oral PV and reindeer PV. The deduced amino acid sequences had strong homologies with the major capsid protein L1 of deer PV, bovine papillomavirus (BPV) 1 and BPV 2, and European elk PV. Although PV antigens were not detected in any of the tumours by immunohistochemistry, PV DNA was demonstrated in individual mesenchymal cells or cell nests of 4/12 tumours by in situ hybridization. A nonproductive infection of mesenchymal fibroblast-like tumour cells with a papillomavirus would explain the lack of PV antigen expression and the absence of PV DNA in the hyperplastic epithelium. Because these tumours and their pathogenesis are similar to equine sarcoids, we suggest that they should be reclassified as 'feline sarcoids' instead of fibropapillomas.     

山羊卵巢无腔卵泡的体外发育

在三维琼脂凝胶培养体系中 ,山羊无腔卵泡的生长模式类似于体内。在一定培养时间内能善为保持其完整的立体结构和形态。原始卵泡能在体外启动 ,并得到一定程度的生长。初级卵泡能发育为次级卵泡 ,次级卵泡发育为有腔卵泡。卵泡在体外发育过程中 ,表现出了明显的优势化。本试验首次初步揭示了山羊无腔卵泡的体外发育基本过程和规律。     

In vitro and in vivo efficacy of lasalocid for treatment of experimental cryptosporidiosis

In vitro viability of purified Cryptosporidium parvum oocysts, exposed for 30, 60, 90 and 120 min to 0.27 mg/ml lasalocid suspension was evaluated by inclusion or exclusion of two fluorogenic vital dyes and an excystation technique. Continuously, preventive and curative efficacies at different doses (9, 6.75, 5.625 and 4.5 mg/kg body weight) and regimens of lasalocid against cryptosporidial infection were evaluated on an experimental neonatal mice model. In vitro assays demonstrated a decrease in the oocyst viability related to an increase in exposure time for exposure to the lasalocid suspension. The infection was eradicated when the suspension was administered with a dose of ≥6.75 mg/kg body weight. No apparent toxic effects were observed.     

一个山羊Ⅰ型毛角蛋白基因的序列及其在皮肤中的表达

角蛋白家族包含表皮软角蛋白和毛发硬角蛋白，可以分为酸性Ⅰ型和碱性或中性的Ⅱ型角蛋白亚家族。从构建的成年山羊皮肤cDNA文库和ESTs分析中发现了和与绵羊羊毛角蛋白8C1和人Ⅰ型毛发角蛋白相应cDNA序列高度同源的序列，经序列分析证明是一个山羊毛发特异性角蛋白，命名为山羊Ⅰ型毛角蛋白1（gHa1），Gen Bank序列号为AY510110．1。推导的读码框ORF由413氨基酸组成，与绵羊8C1角蛋白的序列一致性为97．8％。原位杂交显示它在皮肤初级和次级毛囊的皮质层有强烈的表达。     

白牦牛卵母细胞的采集方法及卵巢贮存条件对其体外成熟的影响

探讨了白牦牛卵母细胞的采集方法以及卵巢保存温度和时间对其体外成熟的影响。结果表明:先抽吸卵泡,每卵巢平均可获可用卵母细胞4.7枚,再切割卵巢,可显著提高可用卵母细胞数9.8枚/卵巢。将屠宰白牦牛卵巢分别置于20~29℃和30~38℃的生理盐水中,6 h内收集卵母细胞,其成熟率、卵裂率分别为65.7%、41.3%和73.3%、45.7%,两者之间的差异均不显著,但均与<20℃组(16.0%、0)差异显著。在30~38℃下,白牦牛卵巢保存时间超过6 h,卵母细胞的成熟率和卵裂率显著下降,卵巢保存时间在2 h以内最好,最晚不超过6 h。     

Voluntary intake of silages in dairy cows depending on chemical composition and in vitro gas production characteristics

A study was conducted to investigate the possibilities to develop models for predicting the relative silage dry matter intake (SDMI) in dairy cows utilising information on chemical composition and in vitro gas production (GP) kinetics of silages. In five experiments, each with an average of 38 lactating dairy cows, SDMI was recorded for 15 grass silages made from primary growth and regrowth swards of timothy (Phleum pratense L.). The silages were characterised by chemical analysis and by utilising an automated in vitro GP recording technique with end point measurements of substrate residues. The silage samples were analysed both as dried and wet samples to evaluate the effects of sample preparation techniques on GP kinetics and their relations to SDMI. Relationships between feed variables and SDMI were investigated utilising simple linear and multiple regression. The wet silage samples had higher cumulative GP and different GP curves compared to the dried samples. The linear relationships between, GP variables, harvest number (first or second cut) of the grass, chemical characteristics of the silages and the relative SDMI show that the GP technique is a powerful tool to detect silage quality. By using the parameters from the dried samples the multiple regression analysis resulted in a relationship, relative SDMI (kg per 100 kg live weight (LW)) per day = 0.071 + 0.0029 × NDFD − 0.266 × C (R² = 0.82, S.D. = 0.06). NDFD is the degradability of the neutral detergent fibre (NDF) (g/kg NDF) and C is the variable that regulates the switching characteristics of the GP profiles. By using the wet silage samples the multiple relationship did not include any GP variable; relative SDMI (kg per 100 kg LW per day) = 1.86 − 0.008 × acetic acid (g/kg DM) + 0.024 × ethanol (g/kg DM) (R² = 0.62, S.D. = 0.08). The results from the regression analysis and the experience of the laborious sample preparation technique for wet samples, give the conclusion that dried silage samples are recommended for determining feed characteristics using the GP technique in intake studies.     

成年发情期奶山羊下丘脑催产素及其mRNA的表达

应用免疫组织化学SP法和原位杂交法研究了催产素（oxytocin，OT）及OT mRNA在成年发情期奶山羊下丘脑中的分布和表达。结果，OT免疫反应阳性细胞主要分布在视上核和室旁核，在视上弥散核、弓状核、室周核和乳头体各核团也存在免疫阳性神经元；在室旁核、视上弥散核、正中隆起和第三脑室附近有较多数量的强阳性神经纤维，在交叉上核有少量阳性神经纤维。在下丘脑23个核团（区）中均能检测出OT mRNA的阳性细胞。结果表明，OT和OT mRNA在下丘脑中分布广泛，且OT可能通过轴突传递和血液运输，将OT mRNA合成的OT运送到别的核团；OT在奶山羊发情过程中发挥了重要作用。     

中链脂肪酸对体外发酵甲烷产量的影响

利用人工瘤胃体外产气法研究不同精粗比(0:1和1:1)日粮类型时中链脂肪酸对甲烷产量的影响。结果显示,体外培养24h,添加脂肪酸对甲烷产量的抑制作用不受日粮类型的影响(P>0.05);与不添加酸的对照相比,添加50mg未酯化月桂酸(C12:0)及月桂酸和肉豆蔻酸(C14:0)以3:2的比例组成的混合物(50mg)可分别抑制甲烷产量的22.2%(P<0.01)和11.8%(P<0.05);添加50 mg未酯化C14:0使甲烷产量降低3.4%(P>0.05);不同日粮类型上饲料产气量、瘤胃液pH值和氨氮浓度差异显著(P<0.01)。     

Low temperature storage and in vitro germination of cherimoya (Annona cherimola Mill.) pollen

Due to the protogynous dichogamy of cherimoya and to the absence of proper pollinating vectors, hand-pollination with fresh pollen is a common practice for cherimoya commercial production. In order to optimize the process of hand-pollination, in this work we have studied the conservation of cherimoya pollen at −20, −80 and −196 °C for up to 3 months. In vitro pollen germination of fresh pollen was 57.1% and it was progressively reduced with conservation time at the three temperatures studied reaching a minimum after 3 months of storage of 10.4%, 14.2% and 13.6% at −20, −80 and −196 °C, respectively. Differences in germination among temperatures were only significant during the first 2 weeks of storage. Field pollinations with pollen stored for up to 3 months at the three temperatures show no yield differences compared to pollinations performed with fresh pollen. The results indicate that pollen collected and stored at sub-zero temperatures at the beginning of the cherimoya blooming season can be used along the whole blooming season avoiding the need of collecting fresh pollen daily.     

梨组织中苹果褪绿叶斑病毒的原位RT-PCR检测

 为了建立梨树苹果褪绿叶斑病毒原位RT-PCR检测技术, 用已知带有苹果褪绿叶斑病毒的库尔勒香梨和无病毒实生苗叶片为材料, 利用D IG标记, 研究了香梨病毒的叶片石蜡切片IS-RT-PCR检测技术。包括AMV逆转录酶、dNTPs、RNasin及互补引物浓度对cDNA合成的影响, 退火温度、Taq DNA聚合酶、Mg²⁺ 、引物浓度及循环次数对原位PCR效果的影响。结果表明: RNasin的用量大于0.2 U·μL^-1时,信号强度随着RNasin量的加大而增强; 只有当dNTPs浓度达1.0 mmol·L^-1时才能生成一定量的cDNA;AMV浓度在0.3～0.5 U·μL^-1均可进行正常的逆转录, 而且在该范围内产物的量随AMV浓度提高而增多; 引物浓度达到0.9μmol·L^-1以上时才能进行有效的逆转录, 并且生成的cDNA的量随引物浓度增大而增加。原位扩增ACLSV的cDNA适合退火温度为56℃。循环20～30次可出现较强的蓝色信号, 引物浓度在0.8～1.2μmol·L^-1时显色较好; Taq酶浓度为20 U·mL^-1以上, 均显示较深的蓝色; Mg^{2 +}浓度为1.5mmol·L^-1就可满足原位PCR所需。获得了苹果褪绿叶斑病毒的原位PCR优化检测体系, 利用建立的优化程序对香梨样品进行了检测验证, 取得了很好效果。