Characterization and virulence of entomopathogenic fungi from sunn pests in Turkey

The sunn pests (Eurygaster spp.) which are important pests of wheat and barley, can lead to 100% yield loss in the case of an epidemic. This study was conducted to characterize the entomopathogenic fungi, obtained from the sunn pests collected from different overwintering areas of Turkey and determine their pathogenicity. Seventeen Beauveria pseudobassiana, 12 Beauveria bassiana, 16 Cordyceps farinosa, 1 Purpureocillium lilacinum, and 2 Clonostachys rosea species were identified. Pathogenicity tests were performed with three B. pseudobassiana, two B. bassiana, four C. farinosa, and one P. lilacinum isolates. B. pseudobassiana isolates BB34 and AF4 caused 100% death on the 9th day, and another B. pseudobassiana isolate E512 caused 100% death on the 12th day. C. farinosa isolate KOKA2 caused 95.8% death on the 15th day. Ultimately, B. pseudobassiana was determined to be the most prevalent and virulent species on sunn pests.     

Entomopathogenic fungi recorded from the harlequin ladybird, Harmonia axyridis

Entomopathogenic fungi were recorded from field samples of the harlequin ladybird Harmonia axyridis, an invasive coccinellid that has recently arrived in Denmark. Larvae, pupae and adults were found to be infected by Isaria farinosa, Beauveria bassiana and species of Lecanicillium. This is the first record of entomopathogenic fungi infecting larvae and pupae. Winter mortality due to fungal infection reached 17.9% in adults collected at one location. The larval stage was most susceptible to fungal infection, as confirmed through bioassay with I. farinosa.     

The entomopathogenic fungi Isaria farinosa (formerly Paecilomyces farinosus) and the Isaria fumosorosea species complex (formerly Paecilomyces fumosoroseus): biology,ecology and use in biological control

The entomopathogenic fungi, Isaria farinosa and Isaria fumosorosea were known as Paecilomyces farinosus and Paecilomyces fumosoroseus, for more than 30 years. Both fungi have a worldwide distribution and a relatively wide host range. While I. farinosa currently is of minor importance in research and as biocontrol agent, I. fumosorosea is regarded as a species complex, and various strains are successfully used for biocontrol of several pest insects, mainly whiteflies. During the past 40 years, numerous scientific papers on both fungal species covering various research areas were published. This is the first monographic review on the biology, ecology and use of I. farinosa and I. fumosorosea as biocontrol agents. In the following compilation, data on the identity, natural occurrence and geographical distribution, host range, production of metabolites, effect of abiotic and biotic factors (temperature, relative humidity, solar radiation, fungus, host plant) and on the use of these species in biocontrol experiments against pest insects, plant pathogens and nematodes in the laboratory and the field are summarised. Sections on their effects on non-target organisms and safety issues are also included. The aim of this review is to increase our knowledge on these entomopathogenic fungi.     

Hypocrealean fungi associated with populations of <Emphasis Type="Italic">Ips typographus</Emphasis> in West Carpathians and selection of local <Emphasis Type="Italic">Beauveria</Emphasis> strains for effective bark beetle control

In Slovakia, a diversity of entomopathogenic fungi (Ascomycota, Hypocreales) associated with outbreaks of Ips typographus was studied in 81 localities and as many as 113 in vitro cultures of five entomopathogenic species were isolated from infected individuals: Beauveria bassiana (87 isolates), B. pseudobassiana (14 isolates), B. caledonica (6 isolates), Lecanicillium lecanii (4 isolates) and Isaria farinosa (2 isolates). B. pseudobassiana is recorded in natural populations of I. typographus for the first time. Biological properties of selected Beauveria isolates, including colony growth, biomass production, conidia yield and pathogenicity to I. typographus adults, were studied in a series of laboratory bioassays and much intra- and interspecific variability was detected. B. bassiana isolates produced biomass or conidia at significantly higher rate than B. pseudobassiana and B. caledonica isolates. Two B. bassiana isolates were selected as the most virulent to bark beetle adults, demonstrating a mean LC₅₀ ranging from 0.72 to 2.05?×?10⁶ conidia ml^?1, and were qualified as promising candidates for biocontrol of I. typographus. Their virulence was significantly higher than that of the mycoinsecticides Boverol®, which was used as a reference strain in the virulence bioassays.     

Isolation and characterisation of Isaria farinosa and Purpureocillium lilacinum associated with emerald ash borer,Agrilus planipennis in Canada

Entomopathogenic fungi of the genera Isaria and Purpureocillium were recovered from infestation sites of emerald ash borer (EAB) in Southern Ontario, Canada. Isolates were identified using morphological characters and by sequencing the ITS1-5.8S-ITS2 ribosomal DNA gene and partial β-tubulin gene. Phylogenetic analysis and constructed trees based on the ITS and β-tubulin gene explicitly confirm isolates L66B, SY17-a and LHY46-a as Isaria farinosa and B3A, B59A and SY45B-a as Purpureocillium lilacinum. Pathogenicity was tested in the laboratory against adult EAB using a single concentration (2.0×10⁷ conidia/ml) applied topically to adults. Controls included three commercial isolates: Isaria fumosorosea LRC176, Metarhizium brunneum LRC187 and Beauveria bassiana strain GHA. The native isolates I. farinosa L66B and P. lilacinum SY45B-a killed 75 and 50% of the beetles 14 days post-inoculation. Although these indigenous entomogenous fungi were less virulent compared with the commercial isolates, yearly isolation from EAB populations suggests they are one of the natural mortality factors of EAB in Canada.     

Toxicity of non-ionic surfactants and interactions with fungal entomopathogens toward Bemisia tabaci biotype B

Several non-ionic surfactants can be used to enhance insecticidal performance of various bio-based products and agrochemicals. In this study, we describe the toxicity of four commercial non-ionic surfactants against immature Bemisia tabaci (Gennadius) biotype B whiteflies along with their spreading ability. Results revealed that trisiloxane-based surfactants (Break-thru^® and Silwet^® L-77) exhibited the highest toxicity to 1st–2nd (early) and 3rd–4th (late) instar nymphs as well as the greatest wetting performance. We also investigated whether sub-lethal concentrations of Break-thru^® and Silwet^® L-77 improved the insecticidal efficacy of the fungal entomopathogens Beauveria bassiana (Balsamo) Vuillemin (strain CG1229) and Isaria fumosorosea Wise (strain CG1228) conidial suspensions against whiteflies. Germination of hydrophobic conidia of both isolates were unaffected by these surfactants over the range 100–1000 ppm. The combinations of both fungi with trisiloxane carriers significantly increased nymphal mortality with mostly additive and synergistic effects on early and late nymphs, respectively. In screenhouse trials, both fungi (10⁷ conidia ml^?1) mixed with 200 ppm of Silwet^® L-77 significantly improved effectiveness against early nymphs (72–74 % mortality) compared with controls treated with water and Silwet^® L-77 alone. In addition, reduced volume rates of I. fumosorosea in Silwet^® L-77 (equivalent to 100 l ha^?1) were at least as effective against late nymphs as higher volume rates (200 l ha^?1) at equivalent conidial doses. Our findings underline the compatibility and enhanced activity of silicon-based surfactants with B. bassiana and I. fumosorosea for use in integrated whitefly management programs.     

An Isaria fumosorosea SCAR marker for evaluation of soil,insect, and airborne samples

Molecular detection markers are needed for ecological studies of entomopathogenic fungi. In this study, a sequence-characterized amplified region (SCAR) marker of Isaria fumosorosea was used to detect the fungus in soil, insects, and airborne samples. These were artificially added with different fungal conidial concentrations. Specificity and sensitivity were tested with semi-nested PCR using oligonucleotides E-AA/M-CTA₁₂₄ F and E-AA/M-CTA₁₂₄ R for the first amplification and E-AA/M-CTA₁₂₄ F and E-AA/M-CTA₁₀₃ R for the second amplification. Specificity assays showed a specific band of 103 bp for DNA samples from 10 I. fumosorosea strains used. Negative results were observed for DNA samples from other species of Isaria, including I. amoene-rosea, I. farinosa, and Paecilomyces carneus as well as with other entomopathogens such as Metarhizium acridum, M. anisopliae, M. majus, M. flavoviride Type E, and Lecanicillium lecanii. Sensitivity assays showed that the specific SCAR marker detected 10⁴ conidia from I. fumosorosea EH-511/3 that were artificially mixed with soil, from 1 to 10⁴ conidia artificially mixed with Galleria mellonella (Lepidoptera: Pyralidae), and from 10 to 10⁵ conidia in Melinex tape for airborne samples. The marker was also able to detect conidia in airborne samples from cotton wicks in a mini wind tunnel. These SCAR markers for I. fumosorosea had excellent specificity and sensitivity and are relevant tools for ecological studies of this fungus.     

Geographic variation in cranial morphology of narwhals (Monodon monoceros) from Greenland and the eastern Canadian Arctic

We analyzed variation in nine non-metric and eight metric variables in the skulls of 132 narwhals (Monodon monoceros) from five localities in Greenland (Inglefield Bredning, Melville Bay, Uummannaq, Disko Bay, and Scoresby Sound) and one in the eastern Canadian Arctic (Eclipse Sound). Metric variables were used to compare the combined Disko Bay and Uummannaq samples with the samples from Inglefield Bredning and Scoresby Sound using three different multivariate techniques for each sex. None of the results were significant. Seven of the non-metric variables were independent of age and sex and were used in comparing samples from the six localities. No differences were found among the four localities in West Greenland, but differences were found in two of the non-metric variables between the combined West Greenland sample and the one from Scoresby Sound. A major shortcoming of the analysis based on metric data was the small sample size from several of the areas, which resulted in low statistical power. Genetic as well as environmental factors could explain the differences detected here between narwhals living along the west and the east coasts of Greenland.     

Effects of three endophytic entomopathogens on sweet sorghum and on the larvae of the stalk borer Sesamia nonagrioides

Various endophytic fungi of native plants and crops are important entomopathogens. The objective of this study was to investigate the entomopathogenic action of Beauveria bassiana (Balsamo) Vuillemin, Metarhizium robertsii (Metchnikoff) Sorokin, and Isaria fumosorosea (Wize) Brown & Smith (all Ascomycota: Hypocreales) against larvae of Sesamia nonagrioides (Lefebvre) (Lepidoptera: Noctuidae) artificially introduced into Sorghum bicolor L. (Moench) (Poaceae) plants under natural environmental conditions. Sorghum bicolor is an economically important crop cultivated for grain, fiber, forage, and lately for biofuel, and S. nonagrioides is its main pest in Mediterranean areas. Young sorghum plants were inoculated with the entomopathogens by spraying in the field. Plant water status, chlorophyll concentration, photosynthesis, and transpiration were not affected. Thirty days after endophyte establishment, plants were infested with fourth instars of S. nonagrioides. The endophytes prevented 50–70% of larvae from entering stalks. Larval mortality was 70–100% and tunnel lengths were reduced by 60–87%. Larval infestation resulted in reduced electron transport capacity and net photosynthetic rate, which was ameliorated in the presence of I. fumosorosea and reversed by B. bassiana and M. robertsii. The growth of sorghum was unaffected in all treatments during the experimental period. Beauveria bassiana and M. robertsii can protect sweet sorghum from damage induced by S. nonagrioides under natural environmental conditions without affecting plant physiology and growth.     

Behavioral and electrophysiological responses of Coptotermes formosanus Shiraki towards entomopathogenic fungal volatiles

Termites adjust their response to entomopathogenic fungi according to the profile of fungal volatile organic compounds (VOCs). This study demonstrates the pathogenicity of Metarhizium anisopliae, Beauveria bassiana and Isaria fumosorosea (=Paecilomyces fumosoroseus) towards the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Using no-choice assays, M. anisopliae was found to be highly virulent (LT₅₀ 3.10 d) when compared to B. bassiana (LT₅₀ 6.62 d) and I. fumosorosea (LT₅₀ 12.39 d). Also using choice assays, the foraging behavior of C. formosanus was determined in the presence of pathogenic fungi. The highly pathogenic fungi (M. anisopliae) elicited a repellent response, causing most of the termites to forage in a safe zone farthest from the fungal source. This repellency resulted in relatively low mortality similar to the controls. The repellency of M. anisopliae conidia can be used to protect human belongings and timber from termites. While I. fumosorosea cultures were not repellent to C. formosanus workers, the termites were highly susceptible to infection. Electroantennographic responses of workers showed approximately 47% and 78% lower level of response to conidia of B. bassiana and I. fumosorosea, respectively, as compared to M. anisopliae. The VOC profile of repellent cultures of M. anisopliae mainly consisted of paraffins (60.97%), while the major proportion of the I. fumosorosea profile consisted of branched and cyclic alkanes (84.41%). From the above findings, we conclude that the incorporation of I. fumosorosea may increase the control potential of bait.     

Efficacy of entomopathogenic hypocrealean fungi against Periplaneta americana

The American cockroach Periplaneta americana, one of the worlds' most important urban insect pests was tested with entomopathogenic fungi. Most promising Metarhizium anisopliae, Metarhizium robertsii and Beauveria bassiana killed nymphs (≥ 81.7% mortality, 25 days after treatment), and these fungi developed on all dead insects. Other fungi tested were less virulent (Metarhizium frigidum and Purpureocillium lilacinum) or avirulent (Isaria cateniobliqua, Isaria farinosa, Simplicillium lanosoniveum, Sporothrix insectorum and Tolypocladium cylindrosporum). Intrageneric and intraspecific variability of fungal activity was detected. Adults were highly susceptible, and oothecae proved to be more resistant than nymphs and adults to infection with M. anisopliae IP 46. Findings of the study underscore the potential of fungi as biocontrol agents against this pest.     

Pathogenicity of fungal isolates (Ascomycota: Hypocreales) against Peregrinus maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae), vectors of corn diseases

Preliminary screening assays were carried out on 17 isolates from five fungal species Beauveria bassiana, Lecanicillium muscarium, Metarhizium anisopliae, Isaria farinosa, and I. fumosorosea. The three most effective isolates against Peregrinus maidis (Hemiptera: Delphacidae) were B. bassiana CEP 147, CEP 150, and CEP 189. There were no consistent differences found in males and females regarding fungal susceptibility. However, more females than males were proportionally infected. There was not a correlation between the percentage of conidial germination and the percentage of mortality caused by fungal infection in any of the treatments. Only B. bassiana CEP 147, which caused a cumulative mortality of 69.8 ± 6.4% after 7 days post-inoculation, was selected to be assayed against adults of P. maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae). In pathogenicity tests significant differences were observed among treatments. After 2 weeks post-inoculation, both D. kuscheli (cumulative mortality of 73.3 ± 9.0%) and P. maidis (cumulative mortality of 68.6 ± 6.7%) were significantly more susceptible than D. maidis (cumulative mortality of 49.9 ± 9.7%) to the selected isolate.     

Compatibility of OMRI-certified surfactants with three entomopathogenic fungi

The Organic Materials Review Institute (OMRI) is a non-profit organisation providing an independent review of products intended for use in organic production systems to certify compliance with US national organic standards. Since all adjuvants to be used in organic agriculture production are required to meet these standards, OMRI's certified list of products is a convenient starting point when developing organic pest control formulations. In the current study, six OMRI-certified surfactants are tested for their compatibility with three common entomopathogens: Beauveria bassiana, Metarhizium brunneum and Isaria fumosorosea. The fungi were evaluated in two common propagule forms, solid-state produced conidia and liquid-media produced blastospores. The results show that most of the surfactants are compatible with the fungi at a high surfactant concentration (2% w/v). In general, the conidia showed a higher susceptibility (greater reduction in spore germination) to the surfactants than the blastospores under these conditions. In addition, the surface tension and foaming properties of the surfactants were determined.     

Entomopathogenic fungi as biological control agents for the vector of the laurel wilt disease,the redbay ambrosia beetle,Xyleborus glabratus (Coleoptera: Curculionidae)

The redbay ambrosia beetle (RAB), Xyleborus glabratus, is a wood-boring insect that vectors the fungal pathogen, Raffaelea lauricola, which causes laurel wilt, a lethal disease of avocado. The objective of this study was to determine the susceptibility of RAB to infection and subsequent death by exposure to three commercial strains of entomopathogenic fungi [two strains of Isaria fumosorosea (Ifr 3581 and PFR), and strain GHA of Beauveria bassiana]. RAB females were dipped in fungal spore solutions and their median survivorship times (MST) determined. Contact with any of the biopesticides resulted in death of all RAB females. MSTs of RAB females ranged from 3 days (B. bassiana) to 5 days (I. fumosorosea PFR). B. bassiana killed RAB females faster, followed by Ifr 3581 and PFR. RAB females dipped in B. bassiana suspensions had the highest number of viable spores attached to their bodies, followed by Ifr 3581. Beetles dipped in PFR suspension had significantly less viable spores attached to their bodies. No significant differences were observed in the mortality of beetles exposed to entomopathogenic fungi by dipping in a fungal suspension or walking on treated avocado bolts. Beetles bored into the logs and constructed galleries, but they were found dead inside the galleries a few days after exposure to the entomopathogens. Entomopathogenic fungal infection in dead beetles was confirmed through molecular techniques. This is the first study to demonstrate that entomopathogenic fungi are potential biological control agents against RAB.     

Effect of Norwegian entomopathogenic fungal isolates against Otiorhynchus sulcatus larvae at low temperatures and persistence in strawberry rhizospheres

Biological control of belowground stages of the black vine weevil Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) in strawberries in cool temperate regions using entomopathogens is challenged by low temperatures during the periods when larvae are vulnerable to infections. In a laboratory study we tested six indigenous Norwegian isolates of entomopathogenic fungi (one Beauveria bassiana, three Beauveria pseudobassiana, and two Metarhizium brunneum; Ascomycota: Hypocreales) for their efficacy against O. sulcatus larvae at 6, 12, and 18 °C. At the lowest temperature only Beauveria spp. affected survival of O. sulcatus while all three fungal species reduced larval survival compared to the control treatment at 12 and 18 °C. Two of the Norwegian isolates, one B. pseudobassiana and one M. brunneum, were then evaluated for long-term persistence (>1 year) in the bulk soil and the rhizosphere soil of strawberries in a semi-field experiment. An exotic isolate of M. brunneum sharing origin with a widespread commercial biocontrol agent (F52/Met52 (Novozymes)) was included for comparison. All three isolates showed significantly higher abundances in the rhizosphere soil compared to bulk soil at 153, 366, and 471 days after inoculation, thus indicating rhizosphere competence for B. pseudobassiana. Notably, CFU levels for both Norwegian isolates were much higher than for the exotic M. brunneum isolate. Selection of locally adapted isolates may therefore be of importance when considering biocontrol strategies of belowground pests in strawberry production.     

Enhanced summer warming reduces fungal decomposer diversity and litter mass loss more strongly in dry than in wet tundra

Many Arctic regions are currently experiencing substantial summer and winter climate changes. Litter decomposition is a fundamental component of ecosystem carbon and nutrient cycles, with fungi being among the primary decomposers. To assess the impacts of seasonal climatic changes on litter fungal communities and their functioning, Betula glandulosa leaf litter was surface‐incubated in two adjacent low Arctic sites with contrasting soil moisture regimes: dry shrub heath and wet sedge tundra at Disko Island, Greenland. At both sites, we investigated the impacts of factorial combinations of enhanced summer warming (using open‐top chambers; OTCs) and deepened snow (using snow fences) on surface litter mass loss, chemistry and fungal decomposer communities after approximately 1 year. Enhanced summer warming significantly restricted litter mass loss by 32% in the dry and 17% in the wet site. Litter moisture content was significantly reduced by summer warming in the dry, but not in the wet site. Likewise, fungal total abundance and diversity were reduced by OTC warming at the dry site, while comparatively modest warming effects were observed in the wet site. These results suggest that increased evapotranspiration in the OTC plots lowered litter moisture content to the point where fungal decomposition activities became inhibited. In contrast, snow addition enhanced fungal abundance in both sites but did not significantly affect litter mass loss rates. Across sites, control plots only shared 15% of their fungal phylotypes, suggesting strong local controls on fungal decomposer community composition. Nevertheless, fungal community functioning (litter decomposition) was negatively affected by warming in both sites. We conclude that although buried soil organic matter decomposition is widely expected to increase with future summer warming, surface litter decay and nutrient turnover rates in both xeric and relatively moist tundra are likely to be significantly restricted by the evaporative drying associated with warmer air temperatures.     

Silica-scaled chrysophytes from West Greenland: Disko Island and the Søndre Strømfjord region

The flora of silica-scaled chrysophytes (Chrysophyceae and Synurophyceae) is known from two previous investigations in southern Greenland, from Narssaq (61°N) and Nuuk (64°N).
In connection with a course in Arctic Biology 25.7.-20.8 1990 chrysophyte samples were obtained from altogether 53 localities, 41 from the Godhavn region in Disko Island (69°N) and 12 from S0ndre Str0mfjord (67°N). The localities were lakes, ponds and puddles, on basalt or an acid bedrock. Fixed samples were examined by EM, and altogether twenty species were identified by their silica scales. Eight of these were new to Greenland, viz. Spiniferomonas silverensis, Mallomonas acaroides, M. alpina, M. crassisquama (previous records proved to belong to M. duerrschmidliae), M. oviformis, M. paludosa, M. tonsurata , and Synura mammillosa. The composition of the flora is discussed from ecological and biogeographic viewpoints.     

Conidial water affinity is an important characteristic for thermotolerance in entomopathogenic fungi

The interspecific thermotolerance of several species of entomopathogenic fungi was evaluated based on the conidial water affinity. The species were divided between hydrophilic and hydrophobic conidia. The species with hydrophobic conidia were Beauveria bassiana (ARSEF 252), Metarhizium brunneum (ARSEF 1187), Metarhizium robertsii (ARSEF 2575), Isaria fumosorosea (ARSEF 3889) and Metarhizium anisopliae s.l. (ARSEF 5749). The species with hydrophilic conidia were Tolypocladium cylindrosporum (ARSEF 3392), Tolypocladium inflatum (ARSEF 4877), Simplicillium lanosoniveum (ARSEF 6430), Lecanicillium aphanocladii (ARSEF 6433), S. lanosoniveum (ARSEF 6651), Aschersonia placenta (ARSEF 7637) and Aschersonia aleyrodis (ARSEF 10276). The conidial surface tension of each isolate was also studied. Conidial suspensions were exposed to 38, 41 or 45 °C. After exposure, the suspensions were inoculated on media and conidial germination was evaluated. Considerable differences in thermotolerance were found among the 12 entomopathogenic fungal species. Species with hydrophobic conidia were generally more thermotolerant than species with hydrophilic conidia. All isolates with hydrophobic conidia showed higher conidial surface tension than the isolates with hydrophilic conidia.     

Soil management effects on entomopathogenic fungi during the transition to organic agriculture in a feed grain rotation

The growing demand for organic products creates opportunities for farmers. Information on the consequences of management practices can help farmers transition to organic and take advantage of these prospects. We examined the interaction between soil disturbance and initial cover crop on naturally occurring entomopathogenic fungi (EPF) during the 3-year transition to organic production in a feed grain rotation in central Pennsylvania. Our experiment included four systems comprised of a factorial combination of two levels of primary tillage (full vs. reduced) and two types of initial cover crop (timothy/clover vs. rye/vetch). The cropping sequence consisted of an initial cover crop, followed by soybean, and finally, maize. The entire experiment was replicated in time, with the initiation lagged by 1 year. We detected four species of EPF (Metarhizium anisopliae, Beauveria bassiana, Isaria fumosorosea, and Isaria farinosa) by bioassay of soil samples collected four times during each field season. The latter three species were detected infrequently; therefore, we focused statistical analysis on M. anisopliae. Detection of M. anisopliae varied across sampling date, year in crop sequence, and experimental start, with no consistent trend across the 3-year transition period. M. anisopliae was isolated more frequently in the systems initiated with timothy/clover cover crops and utilizing full tillage; however, we only observed a tillage effect in one temporal replicate. M. anisopliae detection was negatively associated with soil moisture, organic matter, and zinc, sulfur, and copper concentrations in the soil. This study helps to inform farmers about management effects on soil function, specifically conservation biological control.