Chinese phlebotomine sandflies of the subgenus Adlerius Nitzulescu, 1931 (Diptera: Psychodidae) and the identity of P. sichuanensis Leng & Yin, 1983. II--genotyping and numerical analysis

The authors carried out isoenzyme characterization and genotype comparison studies for Phlebotomus chinensis and Phlebotomus sichuanensis images collected from their holotype/paratype original localities: West Mountain, West Suburb, Beijing and Lixian County, Sichuan Province. Numerical analysis was carried out and the dendrogram of four species of Adlerius was drawn up according to morphological characteristics of these four species. The results showed that P. chinensis and P. sichuanensis can be identified enzymatically in PGM and the MDH-2 allele also differs in P. chinensis from Beijing and. P. sichuanensis. The dendrogram of four Adlerius species showed that P. fengi and P. longiductus are very similar and they clustered first, then P. sichuanensis combine with them to form a combined group. P. chinensis is a separate group from other three. These two groups come to combine at last. Above results have proved that P. sichuanensis is an isolate species instead of a so-called large type P. chinensis.     

Phlebotomus (Adlerius) halepensis vector competence for Leishmania major and Le. tropica

In Eurasia, phlebotomine sandflies of the subgenus Adlerius (Diptera: Psychodidae) comprise about 20 known species. Some are suspected vectors of visceral leishmaniasis (VL) and at least one species has been implicated as a vector of cutaneous leishmaniasis (CL). We tested Phlebotomus (Adlerius) halepensis Theodor (Jordan strain) for CL vector competence, compared with three standard vectors: Phlebotomus (Phlebotomus) duboscqi N-L. from Senegal, Phlebotomus (Paraphlebotomus) sergenti Parrot from Turkey and the Neotropical Lutzomyia longipalpis (L. & N) (Jacobina strain). Sandfly females were membrane-fed on amastigote suspensions of Leishmania major Y. & S. and Le. tropica (Wright) (Kinetoplastida: Trypanosomatidae) and examined for parasite development 3, 6 and 10 days post-infection. Phlebotomus halepensis showed high susceptibility to both leishmanias, supporting typical suprapylarian parasite development similar to the other vectors. Phlebotomus halepensis infection rates were approximately 90% for Le. major and approximately 80% for Le. tropica, with high parasite densities. Development of infections was relatively fast, colonizing the thoracic midgut by 6 days post-bloodmeal in every case and reaching the stomodeal valve in >80% of flies. In late-stage infections, 10 days post-bloodmeal, nearly all P. halepensis females had cardia and stomodeal valve filled with very high numbers of parasites and some Le. tropica-infected females had promastigotes in the pharynx and proboscis. Host choice experiments in the laboratory showed that P. halepensis females fed readily on rat or rabbit and preferred the human forearm. In view of its vector competence and partial anthropophily, we infer that P. halepensis is a potential vector of cutaneous as well as visceral leishmaniases.     

Incrimination of Phlebotomus kandelakii and Phlebotomus balcanicus as vectors of Leishmania infantum in Tbilisi, Georgia

A survey of potential vector sand flies was conducted in the neighboring suburban communities of Vake and Mtatsminda districts in an active focus of visceral Leishmaniasis (VL) in Tbilisi, Georgia. Using light and sticky-paper traps, 1,266 male and 1,179 female sand flies were collected during 2006-2008. Five Phlebotomus species of three subgenera were collected: Phlebotomus balcanicus Theodor and Phlebotomus halepensis Theodor of the subgenus Adlerius; Phlebotomus kandelakii Shchurenkova and Phlebotomus wenyoni Adler and Theodor of the subgenus Larroussius; Phlebotomus sergenti Perfil'ev of the subgenus Paraphlebotomus. Phlebotomus sergenti (35.1%) predominated in Vake, followed by P. kandelakii (33.5%), P. balcanicus (18.9%), P. halepensis (12.2%), and P. wenyoni (0.3%). In Mtatsminda, P. kandelakii (76.8%) comprised over three fourths of collected sand flies, followed by P. sergenti (12.6%), P. balcanicus (5.8%), P. halepensis (3.7%), and P. wenyoni (1.1%). The sand fly season in Georgia is exceptionally short beginning in early June, peaking in July and August, then declining to zero in early September. Of 659 female sand flies examined for Leishmania, 12 (1.8%) specimens without traces of blood were infected including 10 of 535 P. kandelakii (1.9%) and two of 40 P. balcanicus (5.0%). Six isolates were successfully cultured and characterized as Leishmania by PCR. Three isolates from P. kandelakii (2) and P. balcanicus (1) were further identified as L. infantum using sequence alignment of the 70 kDa heat-shock protein gene. Importantly, the sand fly isolates showed a high percent identity (99.8%-99.9%) to human and dog isolates from the same focus, incriminating the two sand fly species as vectors. Blood meal analysis showed that P. kandelakii preferentially feeds on dogs (76%) but also feeds on humans. The abundance, infection rate and feeding behavior of P. kandelakii and the infection rate in P. balcanicus establish these species as vectors in the Tbilisi VL focus.     

Polymerase chain reaction‐based assay for the detection and identification of sand fly gregarines in Lutzomyia longipalpis,a vector of visceral leishmaniasis

Gregarines that parasitise phlebotomine sand flies belong to the genus Psychodiella and, even though they are highly host‐specific, only five species have been described to date. Their most outstanding features include the unique localisation of the oocysts in the accessory glands of the female host, which ensures contamination of the egg surface during oviposition, and the fact that they naturally parasitise the vectors of Leishmania, causal agent of leishmaniasis. The type species, Ps. chagasi, was first described in Lutzomyia longipalpis, vector of visceral leishmaniasis (VL), from Brazil. We recently reported Ps. chagasi sequences in Lu. longipalpis from Posadas (Misiones, Argentina), an endemic VL location where this gregarine had not been previously recorded. In order to analyse the incidence of Ps. chagasi infections in Lu. longipalpis from this location, the aim of this study was to develop a diagnostic assay for sand fly gregarine parasites in Lu. longipalpis. For this, we designed primers using the Ps. chagasi sequences we previously identified and performed an in vitro validation by PCR amplification of the original sand fly samples. Their specificity and sensitivity as diagnostic primers were subsequently confirmed by PCR reactions using total DNA extracted from naturally infected Lu. longipalpis from the same location (Posadas, Argentina).     

Light and Electron Microscopy Studies on the Infection of a Wild‐type and Metalaxyl‐resistant Isolate of Phytophthora sojae in Soybean Hypocotyls

Resistance of soybean cultivars, depending on single dominant genes to Phytophthora sojae, may easily be overcome by emerging new virulent races. Light microscopy (LM) and electron microscopy (EM) were used to study the infection process of the wild‐type isolate Ps411 and metalaxyl‐resistant mutant Ps411‐M of P. sojae in hypocotyls of soybean seedlings grown from untreated and metalaxyl‐treated seeds. The isolate Ps411‐M of P. sojae exhibited a high degree of resistance to metalaxyl compared to Ps411. The pathogenic fitness of Ps411‐M in hypocotyls of soybean seedlings was lower compared to Ps411. LM observations showed distinct differences in the infection process of both isolates in hypocotyls of treated soybean seedlings. EM studies revealed differences in the prepenetration stage between Ps411 and Ps411‐M on hypocotyls grown from seeds treated with 0.02% metalaxyl until the whole seed surface coated. The number of infection sites was markedly reduced and few hyphae continued to spread. Numerous ultrastructural alterations in hyphae were observed in treated hypocotyls infected with Ps411, including pronounced thickening of hyphal cell walls and encasement of haustorium‐like bodies; electron‐dense material was deposited in host cell walls in contact with hyphal cells. Neither the prepenetration process nor penetration or spread of hyphae in the hypocotyls of the resistant isolate was affected in treated compared to non‐treated tissue. While in treated hypocotyls infected with the wild‐type isolate, host defence reactions were induced, no such reactions were detected in treated hypocotyls infected with the resistant isolate. Hypocotyls from metalaxyl‐treated seeds infected with the wild‐type isolate resembled an incompatible interaction, whereas during infection with the metalaxyl‐resistant mutant, the compatible interaction was not changed.     

Species identification and phylogenetic analysis of Leishmania isolated from patients,vectors and hares in the Xinjiang Autonomous Region,The People’s Republic of China

BackgroundVisceral leishmaniasis (VL) has been declared as one of the six major tropical diseases by the World Health Organization. This disease has been successfully controlled in China, except for some areas in the western region, such as the Xinjiang Autonomous Region, where both anthroponotic VL (AVL) and desert type zoonotic VL (DT-ZVL) remain endemic with sporadic epidemics.Methodology/Principal findingsHere, an eleven-year survey (2004–2014) of Leishmania species, encompassing both VL types isolated from patients, sand-fly vectors and Tarim hares (Lepus yarkandensis) from the Xinjiang Autonomous Region was conducted, with a special emphasis on the hares as a potential reservoir animal for DT-ZVL. Key diagnostic genes, ITS1, hsp70 and nagt (encoding N-acetylglucosamine-1-phosphate transferase) were used for phylogenetic analyses, placing all Xinjiang isolates into one clade of the L. donovani complex. Unexpectedly, AVL isolates were found to be closely related to L. infantum, while DT-ZVL isolates were closer to L. donovani. Unrooted parsimony networks of haplotypes for these isolates also revealed their relationship.Conclusions/SignificanceThe above analyses of the DT-ZVL isolates suggested their geographic isolation and independent evolution. The sequence identity of isolates from patients, vectors and the Tarim hares in a single DT-ZVL site provides strong evidence in support of this species as an animal reservoir.     

中国贵州妙蝇属(双翅目,蝇科)研究及净妙蝇群五新种记述(英文)

系贵州地区妙蝇属Myospila Rondani研究的系列论文之一,记述该属1新种群:净妙蝇群M.species-lauta并描述该新种群5新种:冯氏妙蝇M.fengisp.nov.,肖黄基妙蝇M.flavibasisoides sp.nov.,亚黄基妙蝇M.subflavibasis sp.nov.,亚净妙蝇M.sublauta sp.nov.和黄褐妙蝇M.xuthosa sp.nov.。文中给出了净妙蝇群的定义,编制了新种群所有已知种的检索表,描述了新种详细的形态特征及其与近缘种的鉴别特征。新种的模式标本保存于作者所在单位。     

中国秽蝇属一新种 (双翅目：蝇科)

 记述了采自我国四川省的蝇科秽蝇属一新种,冯氏秽蝇Coenosia fengi sp.nov.。新种 具有触角较短、雄蝇阳基后突较弱、肛尾叶和侧尾叶细长等特征,应隶属白角秽蝇种团(C oenosia albicornis\|group)。新种近似于分布在俄罗斯西伯利亚地区的Coenosia ora lis Schnalb,1915,但新种雄蝇侧颜较窄,仅为触角宽的2/5,中胫前背鬃1,后股后腹鬃 仅分布于基部2/3,后胫具2根长的前腹鬃,肛尾叶侧面观较宽等即可区别。模式标本保存于 沈阳师范学院生物系。     

Geographic distribution patterns and status assessment of threatened plants in China

Large scale heterogeneous distribution of biodiversity has become a hot topic for ecologists and conservationists. A threat status assessment combined with geographic distribution patterns of threatened plants in China has been conducted at a national scale in this study based upon a distribution database that refers to both specimen records and published references. Currently, 302 threatened plant species are cataloged in the “National Protected Key Wild Plants” in China belonging to 92 families and 194 genera. Results of the assessment according to the Categories and Criteria system of The World Conservation Union (IUCN) Red List indicate that three species have been assessed as Extinct in Wild (EW) while a further 79, 99 and 112 species have been assessed as Critically Endangered (CR), Endangered (EN), and Vulnerable (VU), respectively. Distribution patterns of threatened plants were analyzed with GIS to identify areas of high species diversity. It was found that threatened plant species occur unevenly within counties and are concentrated in the following eight hotspots: the central and southern Hengduanshan mountain area; the southeast regions of Yunnan as well as Xishuangbanna and southwestern Guangxi; the southern Hainan island; the border mountainous regions of Guizhou, Hunan and Guangxi provinces; the mountainous regions of southwestern Hubei and northern Hunan; southwestern Zhejiang and western Fujian; central Sichuan and southern Gansu; and the western mountains of Guangdong. Moreover, the 12 counties with the greatest number of threatened plant species represent cumulatively more than 50% of the total listed species and, therefore, are the regions in China that should be prioritized for conservation efforts. By overlapping the map of threatened plant species with the distribution of national nature reserves, a gap was identified in protected areas. This research will ultimately provide insights for prioritizing biodiversity conservation as well as processing the mechanisms of distribution patterns.     

Experimental transmission of Leishmania tropica to hyraxes (Procavia capensis) by the bite of Phlebotomus arabicus

The ability of the sand fly Phlebotomus (Adlerius) arabicus to transmit Leishmania tropica was studied experimentally using hyraxes (Procavia capensis), natural reservoir hosts of the parasite. Sand flies became infected with L. tropica after feeding on a lesion of needle-inoculated hyrax. Moreover, P. arabicus fed with L. tropica promastigotes transmitted the parasite to hyraxes by bite during a second bloodmeal. Although the animals remained asymptomatic after infective sand fly bite, they were PCR positive and infectious for naive sand flies. We have thus demonstrated cyclical transmission of L. tropica by P. arabicus in hyraxes. This confirms experimentally the vectorial competence of P. (Adlerius) arabicus, and demonstrates that asymptomatic reservoir hosts are infectious to appropriate vectors.     

Bionomics of phlebotomine sandflies in the Galilee focus of cutaneous leishmaniasis in northern Israel

The bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied for three years (2001-2003) in the Galilee focus of cutaneous leishmaniasis in northern Israel, where the causative Leishmania tropica (Kinetoplastida: Trypanosomatidae) is transmitted by Phlebotomus (Adlerius) arabicus Theodor and Phlebotomus (Paraphlebotomus) sergenti Parrot, comprising 22% and 8%, respectively, of the local sandfly fauna sampled by light traps. The predominant species overall was Phlebotomus (Larroussius) tobbi Adler & Theodor (51%) with lesser numbers of Phlebotomus (Adlerius) simici Theodor (11%), Phlebotomus (Larroussius) syriacus Adler & Theodor (5%), Phlebotomus (Larroussius) perfiliewi Perfil'ev (3%) and Phlebotomus (Phlebotomus) papatasi Scopoli (0.05%). Sandfly adult populations were prevalent from April to November and peaked between June and August, being more abundant through the summer in irrigated habitats, such as gardens and orchards, than in open grassland. Of the two cutaneous leishmaniasis vectors, P. sergenti preferred boulder mounds located at the outskirts of settlements, whereas P. arabicus was more abundant overall and near houses in particular. Females of all these sandfly species displayed a peak of activity after sunset (20.00-22.00 hours), whereas activity of males persisted longer through the night. Another slight increase in activity was noted before dawn (02.00-04.00 hours). Phlebotomus arabicus appears to be the main vector of L. tropica in the Galilee focus, due to its denser populations, more endophily and preference for peridomestic habitats than shown by P. sergenti in northern Israel.     

PANORPIDAE (MECOPTERA) IN XIZANG, CHINA, WITH DESCRIPTION OF THREE NEW SPECIES OF THE GENUS NEOPANORPA

Abstract  Three new species of the genus Neopanorpa Weele are described and illustrated from Xizang in China. The previously reported three species from Xizang, Panorpa guttata Navas, P. davidi Navas, and P. stigmalis Navas, are verified to distribute in Sichuan Province, instead of in Xizang. The type specimens are deposited in Northwest Agricultural University and China Agricultural University separately. A key to the Tibetan species of Neopanorpa is provided.     

Cloning, expression, and purification of a novel recombinant antigen from Leishmania donovani

Visceral leishmaniasis (VL) is a major health problem in the tropical and subtropical regions of the world. The conventional methods for diagnosis of Old World Visceral leishmaniasis are difficult, insensitive, and hazardous. There is no recombinant antigen from old world Leishmania species which can be commercially used for rapid diagnosis. There is an urgent need for a less invasive and accurate method. Here, we report a recombinant antigen from Indian Leishmania donovani for its diagnosis. The kinesin gene of a L. donovani clinical isolate (KE16) from India was PCR amplified for cloning and the immunodominant domain was expressed in Escherichia coli. This recombinant protein or Ld-rKE16 was evaluated for serodiagnosis of Indian kala-azar by ELISA. The recombinant antigen was found to be 100% sensitive and specific for Old World VL cases from India, Pakistan, China, and Turkey. The antigen showed no cross-reactivity with sera from other endemic diseases or healthy controls. The expressed Ld-rKE16 antigen is highly specific and sensitive for diagnosing visceral and post-kala-azar dermal leishmaniasis and is ready for commercialization.     

Trogia venenata (Agaricales), a novel poisonous species which has caused hundreds of deaths in southwestern China

About 400 unexplained sudden deaths have occurred in Yunnan Province in southwestern China during summers over the last 35?years. Extensive field investigations revealed that a fungus called ??little white mushroom?? or ??nail-like mushroom?? by the local people could be one of the chief causes. Fieldwork showed that this fungus is a common saprophytic species growing on rotten wood in southwestern, central, and northwestern Yunnan in subtropical mountainous areas between 1,700 and 3,000?m altitude. Morphological and molecular phylogenetic analyses revealed that it is a new species of the genus Trogia, closely related to the palaeotropical T. infundibuliformis and its allies. Accordingly, T. venenata is described and illustrated.     

Description of a genetic polymorphism of a human proline-rich salivary protein,Pc, and its relationship to other proteins in the salivary protein complex (SPC)

A new polymorphism, Pc, has been identified in human saliva. Two proteins, Pc 1 and Pc 2, are determined by alleles Pc1 and Pc2, respectively, which show autosomal codominant inheritance. No null phenotype has been encountered in 225 randomly collected salivas. The frequencies of the two alleles differ in the Black and White American populations, with Pc1 and Pc2 being 0.670 and 0.330 in the Black (N = 47) and 0.461 and 0.539 in the White (N = 178) populations, respectively. The alleles are in equilibrium in the two populations and segregation analyses (30 families) do not suggest the existence of a null allele in either population. Of seven polymorphic human salivary proteins determined by genes in the salivary protein complex (SPC), Pc phenotypes show association only with Ps phenotypes. Based on that association, our linkage studies, and the biochemical similarities with other SPC proteins, we tentatively conclude that Pc is a member of the SPC, bringing the total number of genes in that group to 13.     

中国前毛广口蝇属分类研究及三新种记述(双翅目：广口蝇科)

对前毛广口蝇属 Prosthiochaeta Enderlein（双翅目：广口蝇科）的属征进行了讨论。详细记述采自我国西南和华中地区的本属3新种: 峨眉前毛广口蝇 P. emeishana sp. Nov.,褐翅前毛广口蝇 P. fuscipennis sp. Nov. 及花翅前毛广口蝇 P. pictipennis sp. Nov.;并附其形态特征图和已知种检索表。 新种模式标本保藏于中国科学院动物研究所。     

西藏新蝎蛉属昆虫三新种记述(长翅目:蝎岭科)(英文)

本文记述中国西藏自治区的新蝎蛉属 Ｎｅｏｐａｎｏｒｐａ Ｗｅｅｌｅ昆虫３新种：西藏新蝎岭 Ｎｅｏｐａｎｏｒｐａｔｉｂｅｔｅｎｓｉｓ Ｈｕａ ｅｔ Ｃｈｏｕ, ｓｐ． ｎｏｖ．（正模,西藏通麦, ２０５０ ｍ）,胡氏新蝎蛉 Ｎｅｏｐａｎｏｒｐａ ｈｕｓｈｅｎｇｃｈａｎｇｉＨｕａ ｅｔ Ｃｈｏｕ, ｓｐ． ｎｏｖ．（正模,西藏樟木口岸）,以及李氏新蝎蛉 Ｎｅｏｐａｎｏｒｐａ ｌｉｆａｓｈｅｎｇｉ　ｈｕａ　ｅｔ　Ｃｈｏｕ,ｓｐ． ｎｏｖ．（正模：,西藏易贡, ２３００ ｍ）。简述了西藏长翅目的分类历史和现状,证实过去记载所谓的三种西藏蝎蛉（模式产地均为马边）,即Ｐａｎｏｒｐａ　ｇｕｔｔａｔａ　Ｎａｖａｓ,Ｐ．ｄａｖｉｄｉ　Ｎａｖａｓ,和Ｐ．ｓｔｉｇｍａｌｉｓ　Ｎａｖａｓ,实际上均为四川省种类。新种模式标本分别保存在西北农业大学昆虫博物馆和中国农业大学昆虫标本室。     

Competitive inhibition of electron donation to photosystem 1 by metal-substituted plastocyanin

The electron transfer from wild-type spinach plastocyanin (Pc) to photosystem 1 has been studied by flash-induced absorption changes at 830 nm. The decay kinetics of photo-oxidized P700 are drastically slower in the presence of Ag(I)-substituted Pc, while addition of Zn(II)-substituted Pc has a weaker effect. The metal-substituted forms of Pc act as competitive inhibitors of the reaction between normal, Cu-containing, Pc and P700. The inhibition constants obtained from an analysis of the kinetic data were 30 and 410 μM for Ag(I)- and Zn(II)-substituted Pc, respectively. When the Gly8Asp mutant form of Pc was used instead of the wild-type form, the corresponding values were found to be 77 and 442 μM. If the Ag- and Zn-derivatives can be considered as structural mimics of reduced and oxidized CuPc, respectively, our results imply that there is a redox-induced decrease in the affinity between Pc and photosystem 1 that follows the electron donation to P700. Our data also imply that the Gly8Asp mutation can diminish the magnitude of this change. The findings reported here are consistent with a reaction mechanism where the electron transfer in the complex between Pc and photosystem 1 is assumed to be reversible.