成都地区四种食尸性蝇类 mtDNA中 COⅠ基因序列检测

通过检测食尸性苍蝇线粒体DNA(mtDNA)上细胞色素氧化酶亚基Ⅰ(COⅠ)中278bp 基因序列,鉴定食尸性苍蝇的种类,解决依据形态学方法不能鉴定苍蝇卵的种类、很难鉴定幼虫种类的难题 ,作为法医鉴别食尸性苍蝇及其幼虫、卵种类依据。随机采集放置在成都地区室外草地兔尸体 上的4 种15个食尸性苍蝇。利用改进的小型昆虫DNA匀浆方法提取上述苍蝇mtDNA;通过Perkin Elmer 9600扩增仪进行PCR扩增;聚丙烯酰胺非变性凝胶连续缓冲体系垂直电泳和银染显色技 术进行扩增结果检测;PCR胶回收试剂盒纯化;ABI 377测序仪测序;MEGA2.1软件包进行序列 分析和构建系统发育树。在双翅目食尸性苍蝇的种内进化分歧均数小于1%,种间进化分歧均数 大于7%。mtDNA上COⅠ序列分析能有效地对主要的食尸性苍蝇进行种类鉴定。该检测方法快速、 简便和精确,能作为法医鉴别食尸性苍蝇种类的可靠依据。     

嗜尸性昆虫墓地栅蚤蝇和柏林克虼蚤蝇的幼期形态及其虫龄鉴别

嗜尸性蚤蝇是掩埋尸体上常见的法医昆虫。利用嗜尸性蚤蝇发育规律推断埋后间隔时间的前提是对其幼期种属的准确鉴定。本文对墓地栅蚤蝇和柏林克虼蚤蝇幼期主要形态特征进行了描述。结果显示:根据口器、节间突起、呼吸角等特征不仅可以区分虫龄,还可以对2种蚤蝇进行鉴别。     

北京地区7种常见嗜尸性蝇类的COI基因序列分析及DNA条形码的建立

嗜尸性蝇类在命案死亡时间和现场推断方面有着十分重要的应用, 而DNA 条形编码技术能摆脱对虫卵和幼虫的饲养以及后续物种鉴定方面专业知识的依赖, 有助于实现现场采集蝇类样本的快速鉴定。本研究采集了北京地区7个嗜尸性蝇类优势种共77个个体的样本, 测定了所有个体线粒体DNA 上细胞色素C氧化酶亚基Ⅰ（COI）基因1 120 bp的序列。基于序列的系统发生分析显示, 同一物种不同个体的序列均以高达99%的支持值聚集在一起。序列间的分歧统计表明这些蝇类在物种内的个体分歧不超过1%, 而不同物种间的净分歧均超过7.74%, 最高可达14.85%。滑动窗口分析表明, 在整个序列区段种间差异位点存在较平均的分布。通过测定COI基因的序列, 建立了北京地区7个嗜尸性蝇类优势种的DNA条形码, 据此实现了对这些物种准确、快速、简单的区分和鉴定, 同时也为后续应用于物种鉴定的种属特异性位点之筛选提供了基础数据。     

吗啡对丝光绿蝇发育积温的改变与死者死亡时间推断的相关研究

用盐酸吗啡注射家兔,处死后用家兔的肌肉组织饲养丝光绿蝇(Lucilia sericata)初孵幼虫,研究吗啡对丝光绿蝇幼虫和蛹生长发育的影响及其在法医学中推断死者死亡时间方面的应用。结果表明,在自然条件下,取食处理组兔肉的丝光绿蝇幼虫的体长和蛹的重量在不同程度上大于对照组;处理组发育积温均比对照组减少。在实验的剂量范围内(4.7~18.8 mg.kg-1),吗啡可促进丝光绿蝇幼虫和蛹的生长发育。根据幼虫或蛹的发育积温推断死者死亡时间,吗啡的这种影响可使推断值产生的最大偏差达80 h左右。     

沈阳地区室内常见尸食性蚤蝇幼期识别

利用尸食性蝇类发育生物学推断死后间隔时间的前提是对其幼期种属的准确鉴定。本文对沈阳地区室内常见的4种尸食性蚤蝇幼期的主要形态特征进行了描述。结果表明：根据其口器、体壁突起、气门和呼吸角等特征不仅可以区分虫龄,还可以鉴别4种蚤蝇。     

湖南省永州地区夏秋季常见嗜尸性昆虫的研究

为了研究湖南省永州地区夏秋季常见嗜尸性昆虫及螺类的种类及其群落演替,运用观察法研究2007—2009年永州地区夏秋季家兔尸上常见嗜尸性昆虫种类变化及其演替。结果表明,该地区家兔尸体上发现部分昆虫纲和腹足纲的嗜尸性昆虫,昆虫纲主要涉及双翅目和鞘翅目的部分昆虫,其中双翅目6科14种,主要有蝇科的孕幼家蝇、斑蹠黑蝇、厚环黑蝇;丽蝇科的丝光绿蝇、叉叶绿蝇、亮绿蝇、大头金蝇和绯颜裸金蝇、白头裸金蝇;麻蝇科的棕尾别麻蝇、酱亚麻蝇;鼓翅蝇科1种;潜蝇科的Phytomyza thalictrella;果蝇科的Drosophila cuaso;鞘翅目5科8种,主要有大黑葬甲、Silpha carinata、Nicrophorus fossor、双色葬甲;蜣螂科的金龟子;步甲科的毛婪步甲和蠋步甲;拟步甲科;隐翅甲科的大黑隐翅虫、小隐翅虫;膜翅目2科3种,蚁科的路舍蚁;胡蜂科的墨胸胡蜂、黑盾胡蜂;鳞翅目的丝兰蛾;腹足纲的钉螺。上述嗜尸性昆虫及螺类在兔尸上出现的时间和部位表现出较强的规律性。研究结果可望为法医在湖南省永州地区对尸体的死亡时间和地点推测提供依据。     

上海地区最常见嗜尸性蝇类三龄幼虫图解检索表

本文提供上海地区最常见嗜尸性蝇类三龄幼虫图解检索表,共4科11种.该检索表可用于案发现场法医昆虫的分类鉴定,进而推断尸体的死亡时间,也可用于重要有瓣蝇类幼虫的孽生地调查.     

我国嗜尸性蝇类的鉴别、发育及演替研究现状与分析

嗜尸性蝇类是法医昆虫学调查中最重要的目标,作为早期到达尸体并主要促成尸体组织腐解的昆虫,对死亡时间的推断,特别是腐败尸体死亡时间的推断具有重大意义。本文针对我国嗜尸性蝇类的主要种类及形态识别、各种类生长发育、在尸体上的演替进行了综述。     

五种常见嗜尸性蝇类的分子鉴定

为了解决法医学人员对于嗜尸性蝇类的鉴定难题,对中山市、广州市及西安市5种常见嗜尸性蝇类共17个样本,对其线粒体COⅠ基因的348 bp大小片段进行了RFLP和DNA序列分析,采用ABI377测序仪测序,DNASTAR软件预测限制性位点,DdeⅠ, DraⅠ和HinfⅠ3种限制性内切酶消化,非变性聚丙烯酰胺凝胶电泳检测酶切结果,MEGA3.0软件包进行序列分析和构建系统发育树。结果表明： 采用mtDNA扩增结合银染技术和DNA序列分析,均可以方便快捷地进行上述三地5种常见嗜尸性蝇类的种类鉴定。本文结果为法医昆虫学中嗜尸性昆虫DNA鉴定数据库的建立提供了数据资料。     

丝光绿蝇不同虫态附着蜡蚧轮枝菌分生孢子的能力及与其体表结构的关系

采用血球计数板计数法测量丝光绿蝇Lucilia sericata成虫、幼虫和蛹三虫态供试虫体体表附着的蜡蚧轮枝菌Verticillium lecanii分生孢子数,并利用扫描电子显微镜等显微技术观察供试虫体体壁外长物分布情况以及附着分生孢子部位的结构形体,来分析丝光绿蝇三种虫态附着蜡蚧轮枝菌分生孢子的能力大小与各虫态体表结构差异的关系。结果表明:蜡蚧轮枝菌分生孢子在丝光绿蝇三种虫态体表的附着数量有显著差异,大小依次为成虫﹥蛹﹥幼虫。成虫周身长有刚毛、鬃等体壁外长物,体壁上密集着生微毛,按其形态结构可将其区分为a和b两种类型;蛹和幼虫的主要体壁外长物是体节上间隔着生并呈"环带状"分布的棘刺,此外蛹端部以及幼虫尾端气门板等区域也着生有体壁外长物,且蛹和幼虫的这些部位的形态结构也存在显著差异。进一步分析后认为,丝光绿蝇各虫态体表附着孢子的能力和体表相关结构的拓扑构型与分布密度等因素相关。本研究可为阐述虫生真菌对同一昆虫不同虫态间的致病差异性的机理提供一定参考。     

Blowfly succession from possum (Trichosurus vulpecula) carrion in a sheep-farming zone

The significance of brushtail possum, Trichosurus vulpecula Kerr (Diprotodontia: Phalangeridae) carcasses to the succession and production of Diptera species and its relevance to fly strike management in Tasmania, Australia was examined. Calliphora stygia (Fabricius), Lucilia sericata (Meigen) and Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) were found to be the most abundant and Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) always the least abundant (< 1%) of the putative primary fly invading species to emerge. Carcasses that were left for up to 15 days in the field before being exposed to flies for 2 days also acted as breeding sites for large numbers of all primary fly species, with the exception of L. cuprina. Ordination analysis revealed no relationship between possum carcasses according to their length of exposure but did show significant negative associations between the number of putative secondary invaders (Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae), Chrysomya varipes (Macquart) (Diptera: Calliphoridae) and putative tertiary flies (Hydrotaea rostrata Robineau-Desvoidy (Diptera: Muscidae)) to the number of C. vicina or C. stygia to emerge. There was enormous variability in the numbers of secondary/tertiary fly species to emerge from carcasses (0-11 450) that negatively correlated with the proportion of all flies to emerge that were primary, and with the mean size of adult L. sericata. Although carcass temperatures, especially those with a large larval population, were elevated, this did not appear to result in significant pre-adult fly mortality. The most important primary fly strike species L. cuprina was only found in insignificant numbers, whereas three other members of the fly strike fauna C. stygia, L. sericata and Ch. rufifacies did use possum carrion as an important breeding resource, but left implications for fly strike management inconclusive.     

A comparison of Lucitraps and sticky targets for sampling the blowfly Lucilia sericata

The Lucitrap (Miazma Pty Ltd, Queensland, Australia) combined with a synthetic odour bait, Lucilure (Miazma Pty Ltd, Queensland, Australia), is a commercially available trap for sampling and control of Lucilia cuprina (Wiedemann) in Australia. It was tested in Hungary against Lucilia sericata (Meigen) (Diptera: Calliphoridae), a cause of sheep strike throughout temperate Europe. The standard Lucitrap was tested against black or yellow sticky target traps. Both trap types were baited with either Lucilure or liver and 10% w/v sodium sulphide solution. With Lucilure as bait, L. sericata were caught on sticky traps but not in Lucitraps. With liver and sodium sulphide as bait, sticky traps caught 500-1500 times more L. sericata than Lucitraps. An adhesive sheet fitted to the top of a Lucitrap captured 30-300 times more L. sericata then were captured inside an unaltered Lucitrap. Direct observation of metallic green calliphorids (92.1% L. sericata) alighting on Lucitraps indicated that most flies stayed for a short while (modal class 2-4 s) and only a few stayed longer, to an observed maximum of 28 s. Flies explored a mean of 1.5 entry holes (range 0-7) during a visit but only 6% entered the trap. Size of L. sericata was not a physical barrier to Lucitrap entry, because many larger species were captured. However, L. sericata captured inside Lucitraps were significantly smaller than those captured on sticky traps, demonstrating that size was of behavioural importance. The data demonstrate that the Lucitrap is not effective as a trap for L. sericata in Hungary, due mainly to a failure of flies to enter the trap in large numbers. In Australia and South Africa, L. sericata is commonly caught in Lucitraps baited with Lucilure, although L. cuprina is more numerous. Our study highlights the potential for diversity of fly behaviour between different geographical populations of the same species. Such diversity can have a significant effect on the functioning of systems for fly sampling and control, when these systems depend for their success on certain behavioural responses of the target species.     

The larvae of some blowflies of medical and veterinary importance

Diagnostic features are described as a series of couplets that enable separation of the third instar larvae of the following pairs of closely related forms of blowflies of medical and veterinary importance: Chrysomya chloropyga (Wiedemann) and Ch.putoria (Wiedemann), Chrysomya albiceps (Wiedemann) and Ch.rufifacies (Macquart), Cochliomyia hominivorax (Coquerel) and Co.macellaria (Fabricius), Lucilia sericata (Mergen) and L. cuprina (Wiedemann), Calliphora augur (Fabricius) and C. stygia (Fabricius).     

Flies in the ointment: a morphological and molecular comparison of Lucilia cuprina and Lucilia sericata (Diptera: Calliphoridae) in South Africa

Abstract.  Complementary nuclear (28S rRNA) and mitochondrial (COI) genes were sequenced from blowflies that phenotypically resembled Lucilia cuprina (W.), Lucilia sericata (Meigen) or exhibited characters of both species. The aim was to test a long-held hypothesis that these species hybridize under natural conditions in South Africa ( Ullyett, 1945 ). Blowflies were obtained predominantly from the Cape Town metropolitan area, but reference samples were acquired for L. sericata from Pretoria. Several L. cuprina -like flies were shown to possess a conflicting combination of nuclear and mitochondrial genes that has also been seen in Hawaiian specimens. Homoplasy, sampling of pseudogenes, hybridization and incomplete lineage sorting are discussed as possible hypotheses for the pattern and the latter is concluded to represent the most likely explanation.     

The role of some cyclorrhaphan flies as carriers of human helminths in Malaysia

The role of some adult flies (Diptera: Cyclorrhapha) as carriers of helminth parasites of man was studied at four sites in Malaysia: a refuse dump, where no helminth-positive flies were detected, and in three peri-domestic situations where four species of flies carried up to three types of nematodes. The dominant fly species Chrysomya megacephala (Fabricius) carried eggs of the roundworm Ascaris lumbricoides L., the pinworm Trichuris trichiura (L.) and hookworm on the adult external body surface and in the gut lumen, in association with Bukit Lanjan aborigines. Chrysomya rufifacies (Macquart) and Sarcophaga spp. also had Ascaris lumbricoides and Trichurus trichiura eggs in their gut contents. Human helminths were not recovered from Lispe leucospila (Wiedemann), Lucilia cuprina (Wiedemann) or the housefly Musca domestica L. In an urban slum area of Kuala Lumpur city, filariform larvae identified as the hookworm Necator americanus (Stiles) occurred in the intestines of the face-fly Musca sorbens Wiedemann (22 larvae per 100 flies) and of Chrysomya megacephala (4.5 larvae per 100 flies). This concentration of apparently infective N. americanus in M. sorbens, a fly which often breeds in faeces and browses on human skin, could have transmission potential.     

LABORATORY STUDIES ON THE CHEMO-SENSORY REACTIONS OF BLOWFLIES

Attraction of various blowflies to sheep wool has been demonstrated under laboratory conditions using a choice-chamber technique. The attractive factor in wool is intimately associated with the wool fibre for it does not disappear during storage and it is not completely removed by washing.
Only female flies were attracted to wool. The highest degree of attraction was obtained from fertilized females which had daily access to meat. Even in the absence of fertilization and meat, attraction was not completely abolished.
The species and strains tested can be placed in the following order in terms of strength of response to sheep wool: Lucilia sericata (British), L. cuprina (tested on damp floor)>L. sericata (Danish: country)> L. sericata (Danish: city), L. sericata (Australian) > L. illustris, L. caesar. No response was given by Calliphora vomitoria.
The olfactory organs involved in the response to wool are mainly on the antennae but antennaeless L. sericata and L. cuprina with some tarsi removed were still able to show some response to wool provided that they were tested on a damp floor. In the course of testing the olfactory responses it was also shown that the main humidity receptors are present on the antennae.
The existence of a 'wool-factor' to which only certain strains of L. sericata and L. cuprina are specially sensitive offers a possible explanation for the greater efficiency of these species as myiasis-producing forms.     

Suppression of populations of Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae), with a novel blowfly trap

The Australian sheep blowfly, Lucilia cuprina , initiates more than 85% of fly strikes on sheep in Australia with an estimated average annual cost of A$280 million to the Australian sheep industry. LuciTrap^® is a commercially available, selective trap for L. cuprina consisting of a plastic bucket with multiple fly entry cones and a synthetic attractant. The impact of LuciTrap on populations of L. cuprina on sheep properties in five Australian states was evaluated by comparing L. cuprina populations on paired properties with and without LuciTraps over seasons when significant fly populations could be expected. Twenty-four comparisons (trials) were conducted over 4 years. During times of 'higher fly density' (when the 48 h geometric mean of trap catches on the control property was greater than five L. cuprina ), the overall geometric mean trap catches for control and trapped properties differed significantly ( P  < 0.001) with mean trap catches of 19.4 and 7.74 L. cuprina , respectively. The selectivity of the LuciTrap was confirmed with 59% of all trapped flies being L. cuprina . Chrysomya spp. and Calliphora spp. constituted 9.3% and 1.1% of the catches with a variety of other flies (mainly Sarcophagidae and Muscidae ) providing the remainder (31%). Lucilia sericata was only trapped in Tasmania and made up 7.7% of the Lucilia spp. catch in that state. Seventy-two per cent of the trapped L. cuprina were female. The deployment of LuciTrap on sheep properties at one trap per 100 sheep from the beginning of the anticipated fly season suppressed the populations of L. cuprina by 60% compared with matched control properties. The LuciTrap is a selective and easy to use fly trap and constitutes an effective, non-insecticidal tool for use in integrated management programs for L. cuprina .     

The molecular systematics of blowflies and screwworm flies (Diptera: Calliphoridae) using 28S rRNA, COX1 and EF-1α: insights into the evolution of dipteran parasitism

The Calliphoridae include some of the most economically significant myiasis-causing flies in the world - blowflies and screwworm flies - with many being notorious for their parasitism of livestock. However, despite more than 50 years of research, key taxonomic relationships within the family remain unresolved. This study utilizes nucleotide sequence data from the protein-coding genes COX1 (mitochondrial) and EF1α (nuclear), and the 28S rRNA (nuclear) gene, from 57 blowfly taxa to improve resolution of key evolutionary relationships within the family Calliphoridae. Bayesian phylogenetic inference was carried out for each single-gene data set, demonstrating significant topological difference between the three gene trees. Nevertheless, all gene trees supported a Calliphorinae-Luciliinae subfamily sister-lineage, with respect to Chrysomyinae. In addition, this study also elucidates the taxonomic and evolutionary status of several less well-studied groups, including the genus Bengalia (either within Calliphoridae or as a separate sister-family), genus Onesia (as a sister-genera to, or sub-genera within, Calliphora), genus Dyscritomyia and Lucilia bufonivora, a specialised parasite of frogs and toads. The occurrence of cross-species hybridisation within Calliphoridae is also further explored, focusing on the two economically significant species Lucilia cuprina and Lucilia sericata. In summary, this study represents the most comprehensive molecular phylogenetic analysis of family Calliphoridae undertaken to date.     

Identification of forensically important blow fly species (Diptera: Calliphoridae) in China by mitochondrial cytochrome oxidase I gene differentiation

Abstract Unambiguous and rapid sarcosaphagous insect species identification is an essential requirement for forensic investigations. Although some insect species are difficult to classify morphologically, they can be effectively identified using molecular methods based on similarity with abundant authenticated reference DNA sequences in local databases. However, local databases are still relatively incomplete in China because of the large land area with distinct regional conditions. In this study, 75 forensically important blow flies were collected from 23 locations in 16 Chinese provinces, and a 278‐bp segment of the cytochrome oxidase subunit I gene of all specimens was successfully sequenced. Phylogenetic analysis of the sequenced segments showed that all Calliphorid specimens were properly assigned into nine species with relatively strong supporting values, thus indicating that the 278‐bp cytochrome oxidase subunit one region is suitable for identification of Calliphorid species. The clear difference between intraspecific threshold and interspecific divergence confirmed the potential of this region for Calliphorid species identification, especially for distinguishing between morphologically similar species. Intraspecific geographic variations were observed in Lucilia sericata (Meigen, 1826) and Lucilia caesar (Linnaeus, 1758).