A mycosis of the blood-sucking snipe fly symphoromyia hirta caused by Erynia ithacensis sp.n. (Entomophthoraceae)

A mycosis was detected in Symphoromyia hirta (Diptera: Rhagionidae) collected near Ithaca, New York. Other dipterous victims of the disease included Rhagio mystaceus and Empis obesa. Afflicted flies, found on the under surfaces of leaves of woody plants, were affixed to the substrate by rhizoids. The pathogen formed both resting spores and conidia on the exterior of the cadavers. It grew rapidly and sporulated abundantly in culture. Attempts to induce infections in Musca domestica and Aedes aegypti were unsuccessful. The pathogen Erynia ithacensis sp. n. forms resting spores that are incised with very irregular ridges. This characteristic serves to separate it from other dipterophilic species of Erynia.This study was supported in part by the UNDP/World Bank/WHO Special Programme for Research in Tropical Diseases.     

The survival ofZoophthora radicans (Zygomycetes: entomophthorales) isolates as hyphal bodies in mummified larvae ofPlutella xylostella (Lep.: Yponomeutidae)

The survival of three isolates ofZoophthora radicans (NW 250, NW 253 & NW 182) as hyphal bodies in dried larvae ofPlutella xylostella stored at 4, 10 and 20°C and 20% R.H was determined. After storage at 20°C, the production of conidia by all isolates was unaffected after 2 weeks but diminished increasingly after 4 and 8 weeks and was entirely lost after 16 weeks. By comparison conidium production at 10°C was unaffected after 16 weeks (isolates NW 250 and NW 182) and, 24 weeks (NW 253) of storage though it declined rapidly in all isolates thereafter. At 4°C many conidia were produced by all isolates even after 34 weeks of storage. These results are consistent with work on other entomophthoralean fungi in dried cadavers suggesting that this may be a common survival strategy in these fungi. NW 250, 253 and 182 were isolated fromP. xylostella in Malaysia and Taiwan, where conditions allow the host to remain active throughout the year. None produced resting sporesin vivo orin vitro but as hosts are always available the ability to survive short dry periods is probably more important than long-term survival for which resting spores are most adapted.      

Susceptibility of different life stages of the European cherry fruit fly,Rhagoletis cerasi,to entomopathogenic fungi

The effects of six fungus isolates on the mortality of different life stages of the European cherry fruit fly, Rhagoletis cerasi (Diptera: Tephritidae), were assessed in a series of laboratory experiments to find an isolate suitable for biological control. In a first step, the effects of fungus treatments on mortality, mycosis and fecundity of adult flies at a concentration of 10⁷ conidia/ml were evaluated. All fungus isolates caused mycosis but virulence varied considerably among the isolates. Beauveria bassiana and Isaria fumosorosea caused 90–100% mortality and had the strongest influence on fecundity. Metarhizium anisopliae also induced high rates of mortality, while the pathogenicity of Isaria farinosa was low. The effects of lower conidia concentrations and the influence of the age of flies were assessed in a second step. Higher conidia concentrations generally resulted in a higher mortality. B. bassiana was most efficient at low concentrations. Young flies showed lower mortality rates than older flies but, sub‐lethal effects on eclosion rate of eggs were greater in younger flies. Finally, the effects on L₃ larvae were tested: none of the fungus isolates induced mortality in more than 25% of larvae. As L₃ larvae and pupae are not susceptible to fungus infection, field control of R. cerasi should be focused on adult flies.     

Isolationin vitro ofStrongwellsea castrans [Fungi: Entomophthorales] a pathogen of adult cabbage root flies,Delia radicum [Dipt.: Anthomyiidae]

The entomogenous fungusStrongwellsea castrans was isolatedin vitro for the first time, by incubating conidia projected from infected cabbage root flies (Delia radicum) in a simple, semi-defined liquid medium comprising dextrose, yeast extract and lactalbumin hydrolysate buffered to pH 7. The fungus grew as long unitunicate hyphae. After transfer to a solid nutrient medium, multinucleate hyphal bodies were formed which developed a thick, laminated wall. Neither conidia nor resting spores developed in liquid or on solid media and the fungus survived successive sub-culturing only in liquid media. Using the API-ZYM system, tests on extracts on hyphae ofS. castrans were positive for 11 enzymes but there were no consistent differences in enzyme profiles betweenS. castrans and fungi of the related genusErynia.      

Development and evaluation of methods to assess populations of black flies (Diptera: Simuliidae) at nests of the endangered whooping crane (Grus americana)

Hematophagous insects can negatively affect the reproductive success of their vertebrate hosts. To determine the influence of hematophagous insects on endangered vertebrates requires specially designed programs that minimize disturbance to the hosts and address problems associated with their small populations. We developed and evaluated a surveillance program for black flies potentially affecting a population of whooping cranes (Grus americana) introduced to central Wisconsin, U.S.A. In one of the few studies to survey host‐seeking female black flies and their immature stages concurrently, we processed nearly 346,000 specimens and documented 26 species, of which only two, Simulium annulus and Simulium johannseni, were attracted to nesting whooping cranes. Attempts to assess black fly populations with artificial nests and real crane eggs were unsuccessful. Carbon‐dioxide traps performed well in describing black fly taxa on the landscape. However, the number of black flies at whooping crane nests was consistently higher than the number captured in carbon‐dioxide traps. The carbon‐dioxide traps poorly described the presence/absence, population fluctuations, and periodicity of black flies at whooping crane nests. The weak performance of the carbon‐dioxide traps might have resulted from microhabitat differences between trap locations and nests or from Simulium annulus and Simulium johannseni using sensory cues in addition to carbon dioxide to find hosts. Choice of trapping techniques, therefore, depends on the information required for the particular study objectives.     

球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响

在室内评价了球孢白僵菌对烟蚜茧蜂生命参数及控害效果的影响。分别在烟蚜茧蜂寄生桃蚜后不同时间进行高剂量（1900孢子/mm^{2）接菌,检测蚜虫感病率和寄生蜂形成的僵蚜率及僵蚜出蜂率。结果表明,球孢白僵菌对僵蚜率和僵蚜出蜂率的影响随接菌时间不同而变化。在烟蚜茧蜂寄生前1d、寄生当天和寄生后3d接菌,蚜虫感病率分别为59.6%、56.2%和34.8%;与对照相比,僵蚜率分别下降94%、59%和47%,僵蚜出蜂率分别减少83%、54%和49%。在寄生后5d或7d接菌,僵蚜率和僵蚜出蜂率不受明显影响,但蚜虫感病率降低到8.2%以下。对蚜尸内白僵菌菌体含量检测表明,随着烟蚜茧蜂寄生后接菌时间的推移,菌体数量迅速下降。寄生蜂寄生后5d或7d接菌,蚜尸内几乎检测不到菌体。直接喷雾接菌烟蚜茧蜂,成蜂寿命缩短4d左右,且81.8%的蜂尸受白僵菌感染。接菌后的寄生蜂对蚜虫寄生率几乎无影响,但寄生蜂在蚜虫体内的存活时间缩短了27.8%。}     

Competitive interaction between strains of Verticillium lecanii on two insect hosts

Differences were observed in the pathogenicities of two strains of Verticillium lecanii (strain numbers 1.72 and 19.79) to the whitefly Trialeurodes uaporariorum and the aphid Macrosiphoniella sanborni. The mean pathogenicity of a mixture of the two strains was intermediate to that of the individual strains, attributable to a competitive interaction during infection. Differences between strains were observed in the production of conidia in vitro and on host insect cadavers. When hosts were infected with the dual-strain suspension, conidia of strain 19.79 only were recovered from T. vaporariorum whereas conidia from both strains were recovered from M. sanborni.     

Effect of Host Plant on the Potential ofPaecilomyces fumosoroseus(Deuteromycotina: Hyphomycetes) for Controlling the Silverleaf Whitefly,Bemisia argentifolii(Homoptera: Aleyrodidae) in Greenhouses

We determined host plant effect on susceptibility of the silverleaf whitefly,Bemisia argentifolii, to the entomopathogenic fungusPaecilomyces fumosoroseus. Whiteflies were reared on three vegetable species (cucumber, cabbage, and tomato) and three cultivars of tomato (Heatwave, Better Boy, and Rutgers). Second instars were sprayed with 5 × 10⁴conidia/cm²ofPfr97, aP. fumosoroseusstrain, used as a microbial control agent of whiteflies. Trials were conducted in an experimental greenhouse, where temperature and relative humidity were adjusted to favor infection (22–33°C, and 68–100% RH). Larval susceptibility to fungal infection was high and not significantly affected by the host plant. Mortality was > 70% 1 week after treatment and increased further during the second week. Percentages of cadavers with subsequent production of conidia observed in the greenhouse did not vary significantly either with the host vegetable species (85–93% 7 days after treatment and 99–100% 14 days after treatment), or with the cultivar of tomato (96–97% 7 days after treatment and 99–100% 14 days after treatment). After incubation under optimal laboratory conditions, the percentages based on the total number of sporulating cadavers (includingin situsporulating individuals and cadavers sporulating afterin vitroincubation) were not significantly influenced either by host vegetable or cultivar of tomato. According to the conditions prevailing in the series of experiments with the three vegetable species or in the series of experiments with the three cultivars of tomatoes, the production of newly formed conidia varied from approximately 10,000 to 18,000 conidia/cadaver. However, in both series, there was no significant influence of the host vegetable species or cultivar. The survival of the newly formed conidia harvested 7 days following treatment reached more than 50% but was not affected by host plant. These results indicate thatP. fumosoroseusshows potential as a microbial control agent for controllingB. argentifoliion greenhouse crops.     

Infectivity of two members of theEntomophthora muscae complex [Zygomycetes: Entomophthorales] forMusca domestica [Dipt.: Muscidae]

Dose-mortality studies were conducted with 2 members of theEntomophthora muscae (Cohn) Fresenius complex from southern California (CA) and Denmark (DA) infecting house flies,Musca domestica L., from southern California. Primary conidia of the DA form were significantly more infective (LC₅₀=34 conidia/mm²) than primary conidia of the CA form (LC₅₀=67 conidia/mm²) for 2–7 days old (‘young’) flies. Infectivity (single experimental trial) for 10–14 day old (‘old’) flies of primary conidia of the CA form (LC₅₀=34 conidia/mm²) was similar to that of the DA form for young flies. Secondary conidia of the CA form were markedly more infective (LC₅₀=0.36 conidia/mm²) than primary conidia of either form. Dose had no significant effect on incubation period for primary conidia of either form, but increased doses resulted in significantly shorter incubation periods for flies exposed to secondary conidia of the CA form.      

Association of melanin content with conidiogenesis in Bipolaris Sorokiniana of barley (Hordeum vulgare L.)

Sixty-seven isolates of Bipolaris sorokiniana of barley, belonging to three groups (black, white and mixed) were studied to find an association of melanin with the spore production of the fungus. Conidiogenesis in black, white and mixed subpopulation of B. sorokiniana was positively correlated with melanin content/g of mycelium. Primary hyphae of black and mixed subpopulation differentiated into secondary hyphal structures which subsequently produced conidiophores and conidia. Primary hyphae could not differentiate into secondary hyphae and subsequently conidiophores and conidia in white subpopulation. A melanin containing mutant developed from white subpopulation regained its ability to differentiate into secondary hyphae, conidiophores and conidia. Results showed that melanization of mycelia B. sorokiniana mycelia is an important factor for conidia production.     

Sporulation by Entomophthora schizophorae (Zygomycetes: Entomophthorales) from housefly cadavers and the persistence of primary conidia at constant temperatures and relative humidities

The duration of discharge of Entomophthora schizophorae (Zygomycetes: Entomophthorales) conidia from house fly (Musca domestica, Diptera) cadavers was measured at 7, 18, and 25 degrees C. The higher the temperature, the shorter the duration of conidia discharge. Significantly more conidia were produced per cadaver at 7 degrees C over a period of 120 h than at 18 and 25 degrees C. At 25 degrees C, the initial discharge over the first 10 h was much larger than at the other temperatures, and at 7 degrees C, no peak in discharge was observed. The persistence of E. schizophorae primary conidia was measured on fabricated non-host surfaces typically found in stables (straw, wood, plaster, and glass) at 7, 18, and 25 degrees C or constant relative humidities of 45, 65, and 85%. Persistence, as measured by the subsequent ability to infect flies, was usually only a few days and depended on the temperature and type of surface. It was greatest on straw, followed by wood, glass, and plaster, and at 7 degrees C, followed by 18 and 25 degrees C. Limited transmission took place between flies exposed to conidia and previously unexposed mates.     

Positive and negative impact to decomposition of pig cadavers by ants (hymenoptera: Formicidae) at the Islet Yeongdo in Busan,South Korea

We observed how ants affected the decomposition process of pig cadavers. Experiments were carried out with six pig cadavers during three seasons without winter. Fifteen ant species belonging to 14 genera 3 subfamilies were recorded from pig cadavers. The species richness and species composition of ants were different significantly at bloated and decay stages from the remaining stages, regardless of season. Ant species Nylanderia flavipes, Aphaenogaster japonica, Pheidole fervida, and Ectomomyrmex javanus kept hunting a considerable amount of eggs and maggots of flies in the two stages. In particular, Lasius japonicus continually interrupted landing and ovipositing of flies and removed eggs and maggots from pig cadavers. Our observation suggests that ants may affect decomposition process of dead animals by participating in positive and negative ways by massive removal of eggs and maggots of flies.     

Effect of exposure method to Beauveria bassiana and conidia concentration on mortality, mycosis, and sporulation in cadavers of Chilo partellus (Lepidoptera: Pyralidae)

The effects of exposure methods, conidial concentrations, and temperature on mortality, mycosis and sporulation in second instar Chilo partellus cadavers infected by Beauveria bassiana was investigated in laboratory studies. Larvae directly sprayed with conidia, exposed to conidia-treated leaves, and dipped into conidial suspension resulted in high mortality (98-100%). The longest LT50 (3.5 days) and days to mortality (2.6 days) were observed in the treated-leaves exposure method. The shortest LT50 (1 day) and days to mortality (1 day) were recorded for the dipping method. With increasing conidial concentrations, there were decreasing LT50 and days to mortality. Larvae exposed to treated leaves and larvae directly sprayed with conidial suspensions produced high mycoses in cadavers. Exposure of larvae to treated-leaves resulted in high sporulation. At lower concentrations of conidia, both mycosis and sporulation in cadavers were high. The optimum temperature for mycosis was 20 and 15 degrees C for sporulation.     

First record of Entomophaga maimaiga (Entomophthorales: Entomophthoraceae) in Slovakia

The entomopathogenic fungus Entomophaga maimaiga was found for the first time in Slovakia in 2013. Late instar larvae of gypsy moth, Lymantria dispar, from two sites with different population densities were dissected to evaluate the presence of pathogens. The presence of conidia and resting spores of E. maimaiga in gypsy moth cadavers was confirmed from both sites.     

Isolation and virulence of entomopathogenic fungi against the great spruce bark beetle,Dendroctonus micans (Kugelann) (Coleoptera: Scolytidae)

Twelve fungal strains including Lecanicillium muscarium (Petch.) Zare and Gams, Isaria farinosa (Holmsk.) Fr., Fusarium sp., Beauveria bassiana Sensu Lato and Beauveria sp. were isolated from larvae and adults of D. micans. In addition, virulence of these isolates against this pest was determined. Conidia suspensions of 1×10⁶ conidia mL^–1 were applied to larvae and adults. The highest mortality and mycosis for larvae were obtained from isolate ARSEF 9271 (Beauveria bassiana) with 90% mortality and mycosis within 10 days. ARSEF 9271 also produced 93% mortality and mycosis in adults. On the other hand, the highest mortality and mycosis for adults were obtained with isolate ARSEF 9272 (Beauveria sp.), with 100% mortality and 80% mycosis within 10 days. These results indicate that isolates ARSEF 9271 and ARSEF 9272 seem to be the most promising potential fungal biocontrol agents against D. micans.     

Bacterial decomposition of insects post-Metarhizium infection: Possible influence on plant growth

Strains of entomopathogenic fungi may have substantial differences in their final stages of mycosis. Insect cadavers are usually overgrown with mycelium after colonization of the insect body, but in many cases, bacterial decomposition of the colonized hosts occurs. We used two Metarhizium robertsii strains in the work: Mak-1 (cadavers become overgrown with mycelium and conidia) and P-72 (cadavers decay after fungal colonization). We conducted a comparative analysis of gut and cadaver microbiota in Colorado potato beetle larvae using 16S rRNA gene sequencing after infection with these strains. In addition, we estimated the content of different forms of nitrogen in cadavers and the influence of cadavers on the growth of Solanum lycopersicum on sand substrates under laboratory conditions. It was shown that infections did not lead to a significant shift in the midgut bacterial communities of infected insects compared to those of untreated insects. Importantly, bacterial communities were similar in both types of cadaver, with predominantly enterobacteria. Decomposing cadavers (P-72) were characterized by increased nitrate and ammonium, and they had a stronger growth-promoting effect on plants compared to cadavers overgrown with mycelium and conidia (Mak-1). We also estimated the colonization and growth of plants after treatment with conidia of both strains cultivated on artificial medium. Both cultures successfully colonized plants, but strain P-72 showed stronger growth promotion than Mak-1. We propose that the use of deviant strains that are unable to sporulate on cadavers leads to a faster (though only passive) flow of nitrogen from killed insects to plants.     

Cues used by the black fly,Simulium annulus,for attraction to the common loon (Gavia immer)

The parasitic relationship between a black fly, Simulium annulus, and the common loon (Gavia immer) has been considered one of the most exclusive relationships between any host species and a black fly species. To test the host specificity of this blood‐feeding insect, we made a series of bird decoy presentations to black flies on loon‐inhabited lakes in northern Wisconsin, U.S.A. To examine the importance of chemical and visual cues for black fly detection of and attraction to hosts, we made decoy presentations with and without chemical cues. Flies attracted to the decoys were collected, identified to species, and quantified. Results showed that S. annulus had a strong preference for common loon visual and chemical cues, although visual cues from Canada geese (Branta canadensis) and mallards (Anas platyrynchos) did attract some flies in significantly smaller numbers.     

Phylogeny of Sibling Species of Simulium venustrum and S. verecundum (Diptera: Simuliidae) Based on Sequences of the Mitochondrial 165 rRNA Gene

The phylogeny of four sibling species of Simulium venustum (CC, CC3, CC4, and AC(gB)) and two sibling species of S. verecundum (AA and ACD = S. rostratum) was reconstructed using nucleotide sequences of the mitochondrial large subunit rRNA gene. Separate phylogenetic analyses were performed by dividing the sequence data into: (1) helices of a computer-generated secondary structure of the rDNA; (2) loops of the computer-generated structure; (3) helices of a consensus secondary structure (deduced by comparing the computer-generated structure of black flies with the corresponding structures proposed for the fruit fly (Drosophila yakuba) and the mosquito (Aedes albopictus)); (4) loops of the consensus structure; (5) both helices and loops of the consensus secondary structure; and (6) the entire sequence regardless of secondary structure, including 11 variable sites in regions where the prediction of secondary structure was not possible. We found that different data sets led to different phylogenetic conclusions. The phylogenies based on data sets 4 and 6 were consistent with nonmolecular evidence, while the phylogenies based on other data sets were not. Our study suggests S. decorum, a morphospecies, might have shared a common ancestor with sibling species of S. venustum.