Species-level Microbiota of Biting Midges and Ticks from Poyang Lake

Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control.Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake.Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors.Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.

Application of the New Gene Gm6 Against Rice GallMidge in Resistance Breeding Through PCR-BasedMarker Aided Selection

The research results of marker aided selection(MAS)for resistant varieties and lines against rice gall midge Orseolia oryzae Wood-Mason successfully in 1999 - 2002 were reported in the present paper. The molecular markers linked to the gene Gm6 against rice gall midge were used to select and breed the resistant varieties and lines. The RAPD marker OPM06 was used to verify the existence actually of gene Gm6 in ten developed varieties resistant to gall midge such as Duokang1, Duokang2, Kangwen2, Kangwen3, Kang-wen5, Duokangzaozhan, Kangwenqinzhan, which were derived from Daqiuqi. For resistance breeding through PCRbased marker aided selection(MAS), the polymorphisms in the resistant and susceptible parents were i-dentified by RG476/Alu I and RG476/Sca I respectively. The RAPD marker OPM06(1.4 kb)was used to i-dentify 15 new resistance lines from F3 lines of Fengyinzhan1/Daqiuqi in 1999. 21 and 7 resistance lines were selected from F4 and F6 lines of KWQZ/Gui99(restored line of hybrid rice)using RG476/Alu I in 2000-2001 respectively. The Gm6 gene was transferred into the restored line of hybrid rice. In 2001 - 2002, RG214/ Hha I and G214/Sca I were used for selecting 11 and 5 resistance lines from F3 lines of KWQZ/IR56 and AXZ/KWQZ successfully. The application of the resistance gene through PCR-based marker aided selection is a new and effective approach in resistance breeding.

Identification and Genetic Analysis of Gall Midge Resistance in Rice Germplasm 91-1A2

Resistance to rice gall midge in rice germplasm 91-1A2 was identified and genetically analyzed F1s of rice population were derived from 91-1A2 which crossed with rice materials Jinggui, TN1, W1263 （Gm1）, IET2911 （Gm2）, BG404-1 （gm3）, OB677 （Gm4）, ARC5984 （Gm5） and Duokang 1 （Gm6） as a male parent. The resistance of all parental lines and F1, BC1F1 and F2 populations to rice gall midge was identified. The results showed that 91-1A2 and all F1s were resistant to Chinese rice gall midge biotype IV. The segregation ratio of resistant plants to susceptible ones in BC1F1 and F2 were accorded with 1：3 and 9：7 rules by X2 test, suggesting that the resistance of 91-1A2 to Chinese rice gall midge biotype IV was controlled by two dominant genes which were new resistance genes, non-allelic to the known rice gall midge resistance genes.

Bluestem Gall Midge Annual Production Cycle and Effects on Grass Seed Production

The bluestem gall midge (Stenodiplosis wattsii Gagné) is native to the grasslands of North America. It feeds on the developing seeds of warm-season grasses during a portion of its lifecycle, but little is known of the biology and extent of gall midge infestations in native warm-season grasses in the USA. We investigated the infestations of the bluestem gall midge in big bluestem (Andropogon gerardii Vitman), sand bluestem (Andropogon gerardii var paucipilus), little bluestem [Schizachyrium scoparium (Michx.) Nash], and indiangrass [Sorghastrum nutans (L.) Nash] commercial seed production fields during 2001 through 2004. Our objectives were to determine the annual production cycle of the bluestem gall midge, the range of the infestation in Nebraska, and estimate its impact on seed production. In eastern Nebraska, the midge goes through four generations with the fourth entering a larval diapause, overwintering in seeds and emerging in mid-June of the following year. The bluestem gall midge damaged from 31% of the little bluestem seed to 67% of the big bluestem seed, and, on average, reduced the production of viable seed by 54% across the four grass species in eastern Nebraska. Additionally, the warm-season prairie grasses were surveyed in nine other counties in Nebraska and the bluestem gall midge was found in each species surveyed from all nine counties. This study is the first to document the negative effects of the bluestem gall midge on the production of viable seed in sand bluestem and indiangrass.

Integrated management of African rice gall midge (AfRGM) (<i>Orseolia oryzivora,</i>Harris and gagné), weeds and diseases of lowland rice

Studies were conducted during the 2001 and 2002 cropping years at Edozhigi and Sachi in the Southern Guinea Savannah of Nigeria to study the effect of following rice with another crop on the next rice crop. There were twelve treatments consisting of poor, good and excellent weed management and pesticidal applications against (AfRGM and diseases set up in a randomized complete block design, in twelve plots in 3 replicates. Results showed that rice treated with gramular isazofos at 0.75 kg a.i. ha–1 and benomyl at 1.0 kg/ha at 20 and 40 days after transplanting (DAT), with good weed management and a follow up crop after rice reduced AfRGM occurrence and disease incidence with a consequent increase in yield of the next rice crop. In 2001 infestation of rice by ARGM was more at Sachi than Edozhigi, while in 2002 there was low ARGM occurrence at the two locations. Brown leaf spot was high at the two locations in the two years of the trial while leaf blast was checked by the integrated control methods used in the study. Higher yields were (1.7 - 11.9 t·ha) from treated-well weeded plots that were cropped to cowpea after the 2001 rice harvest Ludwigia decurens, Sphanoclea geylanica Cyperus difformis and Lepllocidia chinensis were the widely occurring weed species at Edozhigi site while Cyperus difformis, Ipomea mvolucrata, F. awuata and Firnbristylis litoralis were weed species prevalent at Sachi site. The rice nematode, hirschmanniella sp. was recorded at the Sachi site along with other nematodes but was absent at the Edozhigi site. The net benefit farmers would derive for following this rice-dry-season crop sequence vary from US$ 172 - 427 and US$ 175 - 265 ha–1 at Sachi and Edozhigi respectively.

Gas chromatography mass spectrometry based metabolic profiling reveals biomarkers involved in rice-gall midge interactions

The Asian rice gall midge（Orseolia oryzae WoodMason） is a serious pest of rice that causes huge loss in yield.While feeding inside the susceptible host,maggots secrete substances that facilitate the formation of a hollow tube-like structure called gall and prevent panicle formation.The present investigation was carried out to get an account of biochemical changes occurring in the rice plant upon gall midge feeding.Metabolic profiling of host tissues from three rice varieties,namely,TN1,Kavya,and RP2068,exposed to gall midge biotype 1（GMB1）,was carried out using gas chromatography mass spectrometry（GC-MS）.TN1 and GMB1 represented compatible interaction,while Kavya and GMB1 as well as RP2068 and GMB1 represented incompatible interactions.The current study identified several metabolites that could be grouped as resistance,susceptibility,infestation,and host features based on their relative abundance.These may be regarded as biomarkers for insect-plant interaction in general and rice-gall midge interaction in particular.

A Simple DNA Preparation Method for PCR Amplifications in Marker-Assisted Selection of Wheat

An important, but often limiting step in marker-assisted breeding is the efficient isolation of plant DNA for polymerasechain reaction (PCR) amplification. A simple method using an alkali treatment to extract wheat DNA for marker-assistedselection (MAS) in wheat breeding programs was compared to a commercial kit and cetyltrimethylammonium bromide(CTAB) extraction. DNA concentration from the alkali extraction was higher than the other two methods but purity waslower than CTAB extraction. The alkali extraction method was used on breeding lines to determine its usefulness. Thealkali-extracted DNA samples were suitable for several PCR-based procedures, including random amplified polymorphicDNA (RAPD), microsatellite (simple sequence repeat, i.e., SSR) and sequence characterized amplified region (SCAR)analyses.

Evaluation of wheat ear insects in large scale field in central Germany

Wheat ear insects in large scale winter wheat field in Salzmünde (SaxsonyAnhalt) central Germany were evaluated. The present study aimed at studying the abundance of wheat blossom midges WBM, Sitodoplosis mosellana (Géhin), Contarinia tritici (Kirby) and thrips, Li mothrips cerealium (Haliday) and Haplothrips tritici (Kurdjumov). Infestation in winter wheat during the growing seasons 2007, 2008 and 2009 was evaluated. Three methods were used to determine population densities and damage of wheat midges and thrips;pheromone traps, inspection of ear insects and water traps. A strong correlation between midge’s catches and weather conditions was obtained in field ob servations. A positive correlation between phe romone catches and ear infestation levels was recorded;it was higher in 2008 than in 2009. On the other hand, in 2007 there was no synchro nization;S. mosellana hibernated emerged too late to coincide with the susceptible wheat growth stages. The chemical treatment applied at 2008 for highly infestation;there were sig nificant differences in thrips and midge num bers between treated and untreated. Thrips and midge numbers were lower in the treated than in control. The high midge populations in water traps were recorded at growth stages 7779 and 83 and the low populations were recorded at GS 75 and 7577. This gives a reliable base for de cision making to midges control.

Detection of arboviruses in Culicoides(Diptera:Ceratopogonidae)collected from animal farms in the border areas of Yunnan Province,China

Biting midges of the genus Culicoides(order Diptera,family Ceratopogonidae)are potential biological vectors for the transmission of certain arboviruses among humans,livestock,and wild animals.This study collected a total of 405 Culicoides individuals from seven animal farms located in five counties in the border areas of Yunnan Province,China,and examined the Culicoides species composition and the major arboviruses carried by the Culicoides species.The collected Culicoides were classified into seven species with variable abundances:Culicoides arakawae(5.43%,22/405),Culicoides homotomus(1.23%,5/405),Culicoides obsoletus(19.75%,80/405),Culicoides orientalis(17.28%,70/405),Culicoides oxystoma(29.38%,119/405),Culicoides peregrinus(5.68%,23/405),and Culicoides nipponensis(21.23%,86/405).Among the seven species,C.oxystoma and C.nipponensis were distributed in all the five counties with abundances of 13.33–44.87%and 10.00–46.83%,respectively,suggesting that these were the dominant species of Culicoides widespread on animal farms in the border areas.PCR was used to detect major arboviruses in the collected Culicoides specimens,including bluetongue virus(BTV),Japanese encephalitis virus,Dengue virus,Zika virus,African swine fever virus,and African horse sickness virus.Among the tested viruses,only BTV serotype 1 was tested positive in C.oxystoma specimens collected from a buffalo farm.Culicoides oxystoma was the dominant species on animal farms in the sampled areas,but it has not previously been documented as positive for BTV in China.The current results thus suggest that C.oxystoma could be an important vector for BTV transmission in these border areas,which,however,needs to be confirmed by further comprehensive experiments.Overall,the present study provides the first profile of Culicoides species on animal farms in the China,Vietnam,and Myanmar border areas,establishes the prevalence of arboviruses carried by these Culicoides species,and suggests the vector potential of C.oxystoma species for the transmission of BTV.

Differential effects of environmental climatic variables on parasite abundances in blue tit nests during a decade

Models on climate change have predicted an increase of temperature over the earth’s surface with potential drastic effects on living organisms.We analyzed the relationships between climatic conditions(temperature,rainfall,and wind speed)and the abundance of blood-sucking flying insects(biting midges and blackflies)and nest-dwelling ectoparasites(mites,fleas,and blowflies)collected from blue tit nests during bird breeding seasons for a period of 10 years.Average temperature,rainfall,and wind speed showed significant differences among years.Temperature and wind speed increased during the period of study while rainfall decreased.Biting midge,blackfly,and blowfly abundances increased across years but not flea and mite abundances.Hatching date decreased and brood size increased across years.Independently of year variation,parasites were related to climatic variables.For example,biting midge,blowfly,mite,and flea abundances were positive and significantly related to average temperature.We also found a positive and significant relationship between abundances of Haemoproteus infections and biting midge abundances during the first year of life of birds out of nests.However,abundance and prevalence of Lankesterella infections in yearlings were positive and significantly related to mite abundances during the year of birth of birds.Leucocytozoon and Lankesterella infections were also significantly related to climatic variables and Haemoproteus and Lankesterella infections increased across years.In addition,body condition of adult females and males were negatively related to flea larvae and blowfly abundance respectively.Nestling body condition was also negatively related to biting midge abundance.Changes in climatic conditions across years could therefore affect several parasites of birds but also to birds themselves.

Fungi associated with Asphondylia(Diptera:Cecidomyiidae)galls on Sarcocornia quinqueflora and Tecticornia arbuscula(Chenopodiaceae)

Galls induced by the gall-forming midges Asphondylia floriformis and A.sarcocorniae on Sarcocornia quinqueflora,and A.tecticorniae and A.peelei on Tecticornia arbuscula,were collected from two sites near Melbourne,Victoria.Microfungi belonging to a broad range of families were found to be associated with external surfaces of galls and articles of Sarcocornia quinqueflora.However,only Botryosphaeria dothidea was isolated from the fungal mycelium lining gall-midge larval chambers of all four Asphondylia species,on both host plants.