牛皮蝇、纹皮蝇的EST和LDH同工酶的比较研究

本文首次对我国西北地区的牛皮蝇Hypoderma bovis L. 2、3龄幼虫和蛹期与纹皮蝇Hypoderma lineatum(De Villers)的相应阶段的乳酸脱氢酶(LDH)、酯酶(EST)两种同工酶作了聚丙烯酰胺凝胶电泳分析。结果表明两种皮蝇的EST同工酶在酶带数目和相对迁移率(Rf)上均有较大差别;而LDH同工酶仅在2龄幼虫期纹皮蝇比牛皮蝇多显带2条。本文认为EST同工酶对皮蝇属的种和种下分化鉴定有十分重要的意义,而LDH同工酶也可作为鉴定皮蝇属虫种的参考依据。     

纹皮蝇和牛皮蝇的生物学观察

纹皮蝇(Hypoderma lineatum de Villiers)和牛皮蝇(H.bovis de Geer)在我国分布相当广泛,据我们近年来的调查,在东北、华北、内蒙古、西北等地区都普遍发生,其中尤以牧区为严重。这两种皮蝇常在同一地区出现,牛体也同时受感染。在西北甘肃、宁夏等地一般以牛皮蝇数量较多,在内蒙古和华北以纹皮蝇较     

高原鼢鼠寄生皮蝇的形态和分子鉴定

皮蝇蛆病是由双翅目(Diptera)狂蝇科(Oestridae)皮蝇亚科(Hypodermatinae)幼虫感染所导致的一种寄生虫病,会严重影响农牧业发展与公共卫生安全。本研究采用形态学和分子系统学方法对甘南藏族自治州碌曲境内高原鼢鼠(Eospalax baileyi)感染的皮蝇进行鉴定,旨在确定皮蝇的种类,为皮蝇蛆病预防、诊断和控制提供参考。结果发现,高原鼢鼠感染的皮蝇,其三龄幼虫的形态与兔裸皮蝇(Oestromyia leporina)三龄幼虫较为相似;皮蝇三龄幼虫COI基因序列与兔裸皮蝇的差异最小,平均差异为0.63%。系统进化树显示,感染高原鼢鼠的皮蝇三龄幼虫与兔裸皮蝇处于同一进化枝,其亲缘关系最近。因此,本研究确定甘南藏族自治州碌曲境内高原鼢鼠所感染的皮蝇为兔裸皮蝇。     

人体纹皮蝇二龄幼虫的鉴别特征

纹皮蝇Hypoderma lineatum(De Viller,1789)幼虫是人体皮肤蝇蛆症的重要病原,在我国已有不少病例报道。在人体发现的该种蝇蛆均为一、二龄幼虫,文献中对二龄幼虫则描述不详。为了便于人体皮下蝇蛆症的正确诊断,现将我校长海医院皮肤科提供的一例标本与我国常见的牛皮蝇Hypoderma bovis(De Geer,1776)幼虫作比较,供有关工作者参考。 标本来源 自患者皮肤破溃处挤出蝇蛆一条,经鉴定系纹皮蝇二龄幼虫,详细病史及临床资料见另文(陈明、叶琳,1980,皮肤病防治研究通讯(2):26—27)。     

四种麻蝇早期幼虫形态的扫描电镜观察

对常见麻蝇早期幼虫的形态观察研究有助于蝇类早期幼虫的分类及其所致蝇蛆病的诊断。有关蝇类早期幼虫形态的扫描电镜研究报道很少。菊池滋(1972,1974)及作者先后对肠胃蝇(Gasterophilus intestinalis)、牛皮蝇(Hypoderma bovis)、纹皮蝇(H.1ineatum)、羊狂蝇(Oestrus ovis)及7种丽蝇与绿蝇(景涛1985)的早期幼虫与三龄幼虫用扫描电镜作过观察研究。但对麻蝇幼虫的观察至今尚未见详细报道。为此,1981年至1983年对常见4种麻蝇早期幼虫进行了扫描电镜观察。材料与方法标本来源从野外采到怀孕的雌性麻蝇,或用诱蝇笼在旁边开一12×12cm的口,缝…     

青藏高原藏羚羊朊蛋白基因及氨基酸序列分析

羊瘙痒病是一种自然发生的传染性海绵状脑病,它可以在绵羊和山羊羊群之中传播,其疾病的易感性、潜伏期和跨物种传播的能力主要受宿主朊蛋白基因(prion protein gene,PRNP)的影响。研究藏羚羊PRNP有助于明确藏羚羊和羊瘙痒病之间的关系,从而更好地保护藏羚羊。参照GenBank上已经发表的绵羊PRNP序列设计引物,然后对藏羚羊的PRNP序列进行扩增、测序和分析。测序结果显示藏羚羊PRNP序列由771个核苷酸构成,编码256个氨基酸。序列分析结果显示21只藏羚羊PrP(prion protein,PrP)氨基酸序列完全同源,并且与野生型绵羊PrP氨基酸序列完全一致。此外,藏羚羊PrP氨基酸序列与山羊(99.2%)、汤氏瞪羚(99.2%)、印度羚(99.2%)、驯鹿(98%)、马鹿(98%)、家牛(97.7%)和水牛(96.1%)也具有较高的同源性。在对PrP氨基酸序列136、154、171位多态性分析后发现本次收集的21只藏羚羊样本可能和野生型绵羊一样对羊瘙痒病易感。本研究为羊瘙痒病在藏羚羊羊群中可能出现的风险提供了理论依据,提示要加强藏羚羊羊群中羊瘙痒病的监测。     

纹皮蝇(Hypoderma lineatum De Vill.)的形态研究

本文是1957年我们在陕北地区对牛体寄生纹皮蝇进行初步调查观察的结果。在形态学方面,无论是成虫、卵和第一、第三龄幼虫,与以往许多重要文献论著基本上相同;但也有一些是没有记述的,如卵在牛毛上排列的形式,除了有一侧平行整齐排列型和间断分段排列型外,尚有对侧排列型和不整齐排列型。另外,第三龄幼虫的体棘有分叉的现象,即从一个体棘的基部上面分出2—4个小棘;在未成熟第三龄幼虫的节前缘棘的顶端部分较钝。而以上这些特点,尤其是第三龄幼虫体棘的分叉现象,可能为我国纹皮蝇的形态特征之一。     

新疆龙胆科种皮微形态学研究

中国新疆龙胆科5属18种种皮的扫描电镜观察展示了种皮微形态的高度多样性,这5个属可以分为二大类型,第一类包括龙胆属、扁蕾属和獐牙菜属,它们的种皮均具雕纹,此类型又可以分为3小类,每小类对应一个属,这3个属的雕纹类型互不相同;第二类包括假龙胆属和花锚属,它们的种皮均光滑。     

木通科、大血藤科种子的研究，(1)种皮的扫描电镜观察

利用种皮扫描特征进行分类学和系统演化研究,这在木通科,大血藤科研究中尚属 空白。本文就5属15种木通科植物及大血藤科植物的种皮进行了扫描电镜观察。 从种皮雕纹类型看,大血藤科与木通科存在很大差异。Akebia,Holboellia,Stauntonia属植物通常具条纹状种皮雕纹,Sinofranchetia属植物也具条纹状种皮雕纹,但其条纹上具众多疣状凸起, 在 形态上较特殊。Decaisnea属植物的种皮具嵌合型雕纹,这在木通科十分罕见。以上,进一步印证了Stapf等人关于大血藤科分类学处理的正确性,说明了Decaisnea属和Sinofranchetia属在木通科的孤立地位;以及Akebia,Holboellia,Stauntonia属之间较密切的亲缘关系。     

新疆龙胆科肿皮微形态学研究

中国新疆龙胆科５属１８种种皮的扫描电镜观察展示了种皮微形态的高度多样性,这５个属可以分为二大类型,第五类包括龙胆属,扁蕾属和獐牙菜属,它们的种皮均具雕纹,此类型又可以分为３小类,每小类对应一个属,这３个属的雕纹类型互不相同;第二类包括假龙胆 和花锚属,它们的种皮均光滑。     

A third species of Hypoderma (Diptera: Oestridae) affecting cattle and yaks in China: molecular and morphological evidence

Cattle and yak hypodermosis in China is caused by Hypoderma bovis and H. lineatum, with a prevalence reaching up to 98-100% of the animals and maximum intensities exceeding 400 warbles for each animal. A third species, H. sinense, is also considered by Chinese researchers to affect livestock. The molecular characterization of the most variable region of the mitochondrial cytochrome oxidase I gene and of the ribosomal 28S gene has been performed for the third-stage larvae collected from cattle and yaks in China and identified (on the basis of the spinulation on the ventral side of the 10th segment) as H. bovis, H. lineatum, and H. sinense. Amplicons were digested with the HinfI and BfaI restriction enzymes, which provided diagnostic profiles to simultaneously differentiate the 3 Hypoderma species. Third-stage larvae of H. sinense were also examined by scanning electron microscopy, which revealed proper morphological characteristics different from those of H. bovis and H. lineatum. The molecular and morphological evidence herein reported support the existence of a third species of Hypoderma affecting cattle and yaks in China, and the results provide new tools for unequivocal identification of this species and present key components for the evaluation of its endogenous cycle and pathogenicity in animals and humans.     

Shared epitopes between the soluble proteins of Hypoderma lineatum and Hypoderma bovis first instars.

Cattle are known to acquire immunological resistance to hypodermyiasis by repeated exposure to both species of cattle grubs, Hypoderma lineatum (Villers) and Hypoderma bovis (L.). Vaccination of cattle with purified proteins of H. lineatum, particularly hypodermin A, is known to protect cattle against hypodermyiasis by this species. The development of a protective recombinant vaccine against both species using hypodermin A isolated from H. lineatum would require that immunological epitopes be shared by complementary proteins in H. bovis. The purpose of this study was to investigate the soluble proteins of H. bovis first-instars for shared epitopes with H. lineatum. Soluble H. lineatum and H. bovis first-instar larval proteins were resolved by nondenaturing polyacrylamide electrophoresis, blotted onto nitrocellulose paper, and probed with selected polyclonal cow, polyclonal rabbit, and mouse monoclonal antisera. Considerable cross-reactivity was demonstrated by antibodies in the serum of an H. lineatum-infested cow as 6 of 10 resolved H. bovis proteins were bound by the antibodies. The most common shared epitope(s) was associated with hypodermin C, a collagenolytic protease. Hypodermin A shared epitope(s) were noted on 1 prominent H. bovis band (HB1-2). Hypodermin B, a prominent protein in H. lineatum, did not appear to share epitopes with H. bovis proteins. Shared epitopes between H. bovis proteins and hypodermins A and C of H. lineatum would suggest that cross-protection of cattle against H. bovis can be expected by vaccination with recombinant proteins of H. lineatum.     

Scanning electron microscopy of the posterior spiracles of cattle grubs Hypoderma bovis and Hypoderma lineatum

Posterior spiracles of newly hatched first instar larvae of Hypoderma bovis (L.) and H. lineatum (DeVill.) consist of two pairs of spiracular openings. Each pair is surrounded by a rima bearing three spines. Posterior spiracles of second instar larvae are composed of a pair of medial ecdysial scars bounded laterally by spiracular plates. H. bovis spiracular plates have twenty-nine to forty openings, each surrounded by a slightly raised rima. H. lineatum spiracular plates have eighteen to twenty-five openings. Spiracular openings lead to posterior felt chambers which are connected to a common anterior felt chamber filled with a meshlike network. In third instar H. bovis each medial ecdysial scar is surrounded by a strongly concave spiracular plate. Spiracular openings are surrounded by slightly raised rima. Most rimae bear a spine. Spiracular plates of H. lineatum are flat and rimae are without spines. Each spiracular opening leads to a posterior felt chamber, several of which are confluent with a larger anterior felt chamber. Anterior felt chambers open into the dorsal longitudinal tracheal trunk. Felt chambers in third instar larvae are also filled with a complex mesh.     

Early detection of cattle grub (Hypoderma lineatum and H.bovis) (Diptera, Oestridae) using ELISA

An enzyme-linked immunosorbent assay (ELISA) to detect cattle grub-infested animals in the autumn in Alberta has been developed. Antibody to Hypoderma lineatum de Vill. was detectable as early as 6 weeks post-infestation in artificially infested steers. Peak antibody concentrations preceded the peak in maximum 'apparent' grub numbers which occurred between 37 and 43 weeks after infestation. Natural infestations with H. lineatum and H. bovis L., ranging from one to ninety-two grubs per calf, were readily detected by November and the incidence of false positives in uninfested calves was only 5%. Low level (1-4 grubs) infestations of H. bovis only became detectable in February with peak antibody concentrations occurring at the end of April. Prevalence of cattle grub infestations in southern Alberta was shown to be 37% during this study.     

Myiasis caused by Oestridae: serological and molecular diagnosis

Myiasis-causing Oestridae (bot flies) infect several animal species world-wide, from palaearctic to subtropical/tropical areas. Oestrids affect livestock production causing abortion, reduced milk production, losses in weight and fertility, poor hide quality and an impairment of the host's immune system. In the last few years much research has been carried out on the immunology of these infestations, in order to acquire efficient and reliable diagnostic serological tools; the genome of the different species of Oestridae has been studied to further their molecular identification, taxonomy and phylogenesis. The immunodiagnostic methods for many myiasis causing Oestrids have proven to be a viable alternative to the clinical parasitological examination or the post-mortem examination. Numerous serological tests have been developed for the diagnosis of bovine hypodermosis caused by Hypoderma bovis and Hypoderma lineatum, and ELISAs using larval hypodermin C as the antigen are currently used on serum, individual and pooled milk samples to detect the presence of circulating anti-Hypoderma antibodies. In Italy the best period to sample the animals is November-January, since it is in this period that the antibody kinetics of the animals reaches a peak. Recently the efficacy of the ELISA test on pasteurized milk samples has been demonstrated, allowing the diagnosis of bovine hypodermosis also in areas where there is no information on the presence of the disease and the sampling of the animals is laborious. The cross-reactivity between Przhevalskiana silenus antigens and anti-Hypoderma antibodies led to assessing the usefulness of a simple and cost-effective ELISA for the diagnosis of goat warble fly infection. In particular, it has been demonstrated that infected goats display an antibody peak in November-December in blood and milk, thus making this period suitable for sampling. Although no extensive data is available on the immunology of sheep and goat oestrosis caused by Oestrus ovis, the efficacy of ELISA has been demonstrated by correlating serological results with clinical post-mortem examinations. No immunological techniques are currently used to diagnose gasterophilosis of equids and only one study reports the efficacy of ELISA for detecting anti-Gasterophilus antibodies in infected equids. Several studies have been conducted into the molecular characterization of the mitochondrial DNA (mtDNA)--in particular of the gene encoding for the cytochrome oxidase I (COI)--for many free-living and parasitic arthropods for diagnostic, taxonomic and phylogenetic purposes. As regards Oestridae causing myiasis, the first study features a PCR-RFLP assay of the most common Italian species (i.e. H. bovis, H. lineatum, Gasterophilus intestinalis, P. silenus, O. ovis), which showed clear genetic differences among the genera examined, but no inter-specific variation between the two species of Hypoderma considered. The molecular characterization of the most variable region of the COI gene (encoding for the region from E4 to the terminal COOH) was able to clearly differentiate H. bovis and H. lineatum. The E4-COOH region of the COI gene has been characterized for 18 oestrid species and from a taxonomical point of view, molecular data confirm the morphological classification, with the examined species divided into four subfamilies. New insights have also been gained on the molecular differentiation of the most common species of Hypoderma (i.e. H. bovis, H. lineatum, Hypoderma actaeon, Hypoderma diana and Hypoderma tarandi) and, in particular, the restriction enzyme BfaI, provides a diagnostic profile that can be used to simultaneously differentiate all the species examined. The characterization of the E4-COOH COI gene and the hypervariable region of the gene encoding for the ribosomal Isu revealed the identity of Hypoderma sinense as a new species, infecting cattle and yaks in China. Finally, the molecular analysis of the same mitochondrial and ribosomal regions showed that P. silenus, Przhevalskiana aegagri and Przhevalskiana crossii are morphotypes of the same species.     

Species identification of Hypoderma affecting domestic and wild ruminants by morphological and molecular characterization

Cuticular structures and the sequence of the cytochrome oxidase I gene were compared for Hypoderma bovis (Linnaeus), Hypoderma lineatum (De Villers), Hypoderma actaeon Brauer, Hypoderma diana Brauer and Hypoderma tarandi (Linnaeus) (Diptera, Oestridae). Third-stage larvae of each species were examined by scanning electron microscopy revealing differences among species in the pattern and morphology of spines on the cephalic and thoracic segments, by spine patterns on the tenth abdominal segment, and by morphology of the spiracular plates. The morphological approach was supported by the molecular characterization of the most variable region of the cytochrome oxidase I (COI) gene of these species, which was amplified by polymerase chain reaction and analysed. Amplicons were digested with the unique restriction enzyme, BfaI, providing diagnostic profiles able to simultaneously differentiate all Hypoderma species examined. These findings confirm the utility of morphological characters for differentiating the most common Hypoderma larvae and reconfirm the power of the COI gene for studying insect identification and systematics.     

Comparative scanning electron microscopy of third-instar Hypoderma spp. (Diptera: Oestridae)

Scanning electron microscope study of third-instar larvae of four species of Hypoderma revealed differences among species in the pattern of spination, spine morphology and morphology of the spiracular plates. These observations identify characters that enable the differentiation of Hypoderma actaeon and H. diana, parasitizing red deer ( Cervus elaphus ) in Europe, and provide additional characters for differentiating H. bovis and H. lineatum parasitizing cattle.     

皮下盘菌属的两个新分类单元

报道中国黄山产皮下盘菌属二新种:生于满山红Rhododendron mariesii叶片上的满山红皮下盘菌Hypoderma rhododendri-mariesii sp. nov.和生于野珠兰Stephanandra chenensis枝干上的野珠兰皮下盘菌H. stephanandrae sp. nov.。对它们作了拉丁文特征简介、汉文描述和图解。模式标本存放在安徽农业大学森林保护教研室(AAUFP),合肥。