青海省藏羊感染皮蝇蛆调查及其虫种鉴定

为了解青海省藏羊感染皮蝇蛆状况及感染种类,本研究在3个地区调查了476只藏羊,对2株感染藏羊皮蝇蛆和4株感染牦牛皮蝇蛆的线粒体CO1基因种属特异性序列进行比较研究。结果表明:14只羊感染了皮蝇蛆,感染率为2.9%,总瘤胞数为27个,平均感染强度为1.9个;并扩增其长度为688 bp的序列,克隆后测序,将其序列与GenBank中的牛皮蝇、纹皮蝇蛆和中华皮蝇蛆序列进行聚类分析,结果显示青海省感染藏羊的皮蝇蛆与感染牦牛的果洛株、民和和黄南株在线粒体CO1基因片段聚类分析进化树和同源性比较中,与牛皮蝇位于同一个进化树分支上,基因片段同源性大于99%,为牛皮蝇;感染牦牛的海晏株和贵南株在线粒体CO1基因片段聚类分析进化树和同源性比较中,与中华皮蝇位于同一个进化分支上,基因片段同源性大于99%,为中华皮蝇。几个皮蝇蛆分离株种内变异率小于1%,种间变异率大于8%。     

牛皮蝇幼虫病的流行及防制

牛皮蝇幼虫病是一种常见的寄生虫，在我国许多地区都有发生，引起牛的生产性能下降，造成经济上的损失。本文主要阐述病原体形态特征、生活史、流行规律、危害及诊断方法，并提出综合防制措施。     

感染高原鼠兔的皮蝇蛆16SrRNA基因序列研究

利用分子生物学技术对高原鼠兔感染皮蝇蛆16SrRNA基因进行了研究,扩增出长度为551 bp的序列,克隆测序,将其序列与GenBank中的皮蝇科、胃蝇科的序列进行了聚类分析,并构建分子进化树.结果显示,感染高原鼠兔的皮蝇蛆与皮蝇科的皮蝇蛆遗传距离更近,在一个大的进化树分支上,基因片段同源性84.6％～88.4％之间,与胃蝇科的红尾胃蝇基因片段同源性60.1％ ～64.4％之间.几株感染高原鼠兔的皮蝇蛆之间的基因片段同源性94.4％～99.6％之间.     

鹿皮蝇蛆对驯鹿抗氧化能力的影响

为了探讨驯鹿鹿皮蝇蛆对驯鹿机体氧化应激的影响,分别对鹿皮蝇蛆感染组和未感染组驯鹿血清中的总超氧化物歧化酶(T-SOD)、一氧化氮(NO)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、羟自由基(.OH)及总抗氧化能力(T-AOC)进行测定。结果表明鹿皮蝇蛆感染组血清中T-SOD、T-AOC活性、抑制.OH能力及NO含量极显著低于未感染组(P<0.01),血清中GSH-Px活性显著低于未感染组(P<0.05);血清中MDA含量极显著高于未感染组(P<0.01)。说明驯鹿感染鹿皮蝇蛆后,机体清除自由基能力降低,自由基在机体内大量蓄积,导致机体产生氧化应激反应。     

埃普利诺菌素浇泼剂对牦牛皮蝇幼虫驱除效果试验

应用埃普利诺菌素浇泼剂驱除牦牛皮蝇幼虫,并设埃普利诺菌素注射剂药物对照组和阳性对照组。结果：剖检埃普利诺菌素浇泼剂300μg/kg,400μg/kg,500μg/kg剂量组牦牛与药物对照埃普利诺菌素注射剂组及伊维菌素浇泼剂组牦牛,均未检出皮蝇1期幼虫,而阳性对照组牦牛皮蝇1期幼虫感染率35.0%,平均感染强度29.4（9～53）条。翌年3、5月份两次触摸检查牛背部皮下瘤疤和皮肤虫孔,4个药物试验组牦牛均未检出,而阳性对照组牦牛两次检查平均感染率45.16%,平均感染强度6.45个。试验证明埃普利诺菌素浇泼剂低、中、高3个剂量对寄生于牦牛的皮蝇幼虫驱净率、驱虫率均达100.0%,高效安全。     

Hypodermosis in cattle slaughtered in Nigde province,Turkey<Superscript>*</Superscript>

This study was carried out to investigate the prevalence of hypodermosis in cattle between January and June 2005 in Nigde province, which is located in the middle of Turkey. A total of 1336 cattle, which were slaughtered in Nigde, were investigated for Hypoderma larvae. The 68 out of 1336 cattle (%5.08) were found positive for Hypoderma larvae. A total of 536 Hypoderma larvae were found in the skin and subcutaneous tissue of the back of infested cattle. The 489 out of 536 larvae (%91.23) were identified as Hypoderma bovis and 47 out of 536 (%8.77) as H. lineatum. Number of Hypoderma larvae counted on single infested cattle varied between 1–45 and the mean number of Hypoderma larvae per cattle was 7.88 (536/68). Hypodermosis was recorded for the first time in cattle from Nigde. ^*This study was supported by the Scientific Research Projects Unit of Nigde University (FEB 2004/07).     

Local and systemic cytokine responses during larval penetration in cattle experimentally infested with Hypoderma lineatum (Diptera: Oestridae)

Local and systemic cytokine responses were studied in 3 groups of cattle, with 4 animals each, experimentally infested with Hypoderma lineatum (De Villers) first instars (L1). The first group was undergoing a primary infestation (G-1), the second group was undergoing a secondary infestation (G-2) and the third group was infested for their third consecutive year (G-3). Cattle were infested with 25 L1 deposited on the skin. Blood and skin samples were taken at 0, 6, 12, 48, 96 and 144 h post-infestation (h.p.i.). Interleukin 10 (IL-10), IL-4 and interferon gamma (IFN-γ) production was studied by immunohistochemistry and sandwich ELISAs. IL-4⁺ cells showed a significant increase at 6 h.p.i. in both reinfested groups (G-2 and G-3) when compared with G-1. In all groups the number of IL-4⁺ cells decreased significantly at 48 h.p.i. IL-10⁺ cells increased in G-1 at 6 and 48 h.p.i., whereas in both reinfested groups increased at 12 h.p.i. with a peak at 48 h.p.i. IFN-γ⁺ cells showed a significant increment at 6 h.p.i. in all groups, followed by a rapid descent at 12 (G-1 and G-2) and 48 h.p.i. (G-3). Penetration of the skin by H. lineatum did not have any significant effect on IFN-γ serum concentrations and, except for IL-10 there were no correlation between local production and serum concentrations of cytokines. The increase of both Th1 (IFN-γ) and Th2-type cytokines (IL-4 and IL-10) indicates that bovine T-cell response during the first phases of the infestation by H. lineatum is apparently a Th0 response.     

皮蝇素A基因的克隆与序列分析

首先提取采自甘肃玛曲的皮蝇一期幼虫的总RNA,合成cDNA,根据设计的特异引物,利用PCR技术扩增出Hypodermin A(HA)基因片段,将此片段与PGEM-T Easy载体连接,构建克隆载体PGEM-HA,转化大肠杆菌JM109,筛选阳性克隆,进行测序与分析。结果表明,扩增的HA基因长678bp,编码229氨基酸。克隆到的基因序列与NCBI GenBank上登陆的皮蝇HA基因序列核甘酸同源性为96.6%,推导氨基酸同源性达97.8%。该基因的成功克隆为其进一步表达打下了坚实的基础。     

纹皮蝇Hypodermin C基因原核表达及间接ELISA检测方法的建立

为建立牛皮蝇蛆病的抗体检测方法,本研究从纹皮蝇I期幼虫中提取总RNA,采用RT-PCR扩增了Hypodermin C(HC)基因.将该基因片段插入到原核表达载体pET-30a(+)中进行原核表达.获得了31ku以包涵体形式表达的重组HC蛋白.Western blot结果表明表达产物能够与阳性牛血清IgG特异性结合,具有良好的抗原活性.以纯化的重组HC作为包被抗原建立了牛皮蝇蛆病间接ELISA诊断方法,实验确定抗原最佳包被浓度为15.63 μg/mL,血清最佳稀释度为1:80,阳性标准判定为待检血清OD值＞0.256.所建立的诊断方法具有较好的特异性、敏感性和可重复性.应用该检测方法对230份临床血清进行检测,阳性率为20.6％.研究结果表明该间接ELISA方法是一种有效的牛皮蝇蛆病血清学检测方法.     

The efficacy of eprinomectin extended-release injection against Hypoderma spp. (Diptera: Oestridae) in cattle

The efficacy of eprinomectin in an extended-release injection (ERI) formulation was determined in cattle harboring naturally acquired infestations of first- or second- and third-stage larvae of Hypoderma spp. in three studies conducted according to the same protocol in the USA (two studies) and Germany (one study). Thirty cattle sourced from herds with a history of Hypoderma infestation were included in each study. Cattle were formed into replicates of three animals each on the basis of pre-treatment anti-Hypoderma antibody titers. Within replicates each animal was randomly allocated to one of the following treatments: ERI vehicle (control) at 1 mL/50 kg bodyweight, administered once on Day 0; Eprinomectin 5% ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg), administered once on Day 0 (when larvae were expected to be first instars); or Eprinomectin 5% ERI at 1 mL/50 kg bodyweight (1.0 mg eprinomectin/kg), administered once when larvae were second or third instars (study dependent, Day 73, 119, or 140). Treatments were administered by subcutaneous injection in front of the shoulder. In all studies, emerging and/or expressed Hypoderma larvae were recovered, speciated, and counted and viability was determined. Eprinomectin LAI treatment was 100% (p < 0.05) efficacious against first- and second- or third-stage larvae of Hypoderma bovis (two studies) and Hypoderma lineatum (one study). All animals accepted the treatment well. No adverse reaction to treatments was observed in any animal in any study.