Pine processionary moth (Thaumetopoea pityocampa,Lepidoptera: Thaumetopoeidae) larvae are highly susceptible to the entomopathogenic fungi Metarhizium brunneum and Beauveria bassiana

The larvae of the pine processionary moth (PPM), Thaumetopoea pityocampa, feed on the needles of pine and cedar. The urticating hairs of older instars pose a threat to human and animal health. Strains of the entomopathogenic fungi, Metarhizium brunneum (V275, ARSEF 4556) and Beauveria bassiana (KTU-24), were assayed against first to fourth instar T. pityocampa using doses ranging from 1?×?10⁵ to 1?×?10⁸ conidia mL^?1. The three strains differed slightly in their virulence but caused 100% mortality of all instars at the highest dose. The newly emerged or first instar larvae were extremely susceptible with 100% mortality being achieved 2–4 days post inoculation with V275 at all but the lowest dose. The fourth instar larvae appeared to be less susceptible than earlier instars. There was good horizontal transmission of conidia from treated to un-inoculated larvae. However, mortality was higher in third and fourth instars and where the ratio of inoculated versus untreated larvae was high. This we presume is due to spores being more readily trapped by the urticating hairs found on third and older instar larvae. Injection of the nests offers a simple and environmentally friendly way of controlling the pest with reduced risk to operators.     

Virulence of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against red palm weevil,Rhynchophorus ferrugineus (Olivier)

Nineteen different isolates of the entomopathogenic fungi Beauveria bassiana sensu lato (s.l.) and Metarhizium anisopliae s.l. (Ascomycota: Hypocreales), recovered from different soil samples (field crops, fruit orchards, vegetable fields and forests) and insect cadavers were tested against Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) at two different spore concentrations (1 × 10⁷ and 1 × 10⁸ conidia mL^?1). Three isolates of B. bassiana and two of M. anisopliae gave >88 % larval and >75 % adult mortality of R. ferrugineus on their highest dose rate respectively. More sporulating cadavers (mycosis) resulted from a high dose rate compared to low dose on both life stages of R. ferrugineus. The current study confirmed the lethal action of B. bassiana and M. anisopliae isolates with mortality levels usually directly proportional to the conidial concentration. This study further confirmed that the isolates recovered from R. ferrugineus dead cadavers gave more mortality compared to the other sources. In the virulence assay two isolates of B. bassiana caused the highest percentage of both larval and adult mortality at all exposure intervals which suggest that they may be the most promising for use in sustainable management programs aimed at microbial control in date palm orchards.     

球孢白僵菌和金龟子绿僵菌不同菌株对黑足角胸叶甲成虫的致病力评价

本研究在测定不同球孢白僵菌Beauveria bassiana和金龟子绿僵菌Metarhizium anisopliae菌株的生长速率与产孢量的基础上, 采用孢悬液浸渍法进行了对油茶新害虫--黑足角胸叶甲Basilepta melanopus成虫的生物测定, 旨在筛选出感染该成虫的高致病力菌株, 为防治该虫提供新的生物资源。结果表明： 不同菌株生长速率和产孢量间存在显著差异, MaYTTR-04, BbFZ-17, MaZPTR-01和BbTK-01生长速率和产孢量均显著高于其他菌株。接种后, 叶甲成虫累积死亡率随时间的增加而逐渐增高, 接种白僵菌7 d后, 成虫校正死亡率全部达到100%; 接种MaZPTR-01和MaYTTR-04两个绿僵菌菌株14 d后, 成虫死亡率分别为80.3%和78.8%。而且接种白僵菌后, 叶甲成虫的僵虫率显著较绿僵菌高, 尤其以BbTK-01和BbFZ-17两个菌株较好, 分别达到85.7%和75.8%。BbXJ-01, BbFZ-17和BbTK-01 3个白僵菌菌株的LT₅₀最小, 分别为3.0, 3.3和3.4 d; MaYTTR-04和MaZPTR-01两个绿僵菌的LT₅₀分别为6.0和6.2 d。结果说明, 白僵菌对叶甲成虫的致病力较强, 尤其是BbTK-01和BbFZ-17两个菌株, 不仅致死率高, 且致死速度快, 僵虫率高, 同时这2个菌株生长速度快、 产孢量大, 具有优良的生产特性, 在黑足角胸叶甲的生物防治中将有重要的应用价值。     

Susceptibility of apple clearwing moth larvae,Synanthedon myopaeformis (Lepidoptera: Sesiidae) to Beauveria bassiana and Metarhizium brunneum

Apple clearwing moth larvae, Synanthedon myopaeformis (Lepidoptera: Sesiidae) were found to be susceptible to infection by two entomopathogenic fungi: an indigenous fungus isolated from S. myopaeformis cadavers and identified as Metarhizium brunneum (Petch); and Beauveria bassiana isolate GHA. In laboratory bioassays, larvae exhibited dose related mortality after exposure to both the M. brunneum and Beauveria bassiana with 7 day LC₅₀'s of 2.9×10⁵ and 3.4×10⁵ spores/mL, respectively. Larval mortalities caused by the two isolates at 1×10⁶ spores/mL were not significantly different and 73% of the M. brunneum-treated, and 76% of the B. bassiana-treated larvae were dead 7 days post treatment, with LT₅₀'s of 5.5 and 5.1 days, respectively.     

Persistence of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato in Vermont (USA) forest soil

Persistence of granular formulations of Beauveria bassiana sensu lato, strains GHA726 and ERL1170, and Metarhizium anisopliae sensu lato, strains ERL824 and ERL701, applied to forest soil was documented. One formulation was prepared by producing fungi on millet and at application adding cooked, dried millet to provide additional nutrients for fungal growth. The other was made by producing fungi on corn starch ‘packing peanuts’, a biodegradable packaging material, adding additional packing peanuts as a nutrient base. The trial was conducted in three sites in a forest predominating in sugar maple (Acer saccharum) in Underhill, VT, USA. In each site, 33 dominant sugar maples were randomly selected. A 1-m² plot located 1 m from the bole of the tree was established. Soil samples were collected before application, and 1 h, 2, 4, 8, 12 and 16 wk post-application. Fungal levels were estimated based on number of colony-forming units (CFUs). A significant increase in CFUs was observed 2 wk post-application in plots treated with isolates GHA726, ERL1170 and ERL824 as a result of fungal reproduction on nutrients in the formulations. Evidence of greater persistence for the B. bassiana than the M. anisopliae isolates was observed, based on CFU levels over time. Isolates formulated with millet proliferated more than those on packing peanuts. Use of entomopathogenic fungi in forest soil has potential for managing soil-dwelling insect pests such as pear thrips, Taeniothrips inconsequens, and the black vine weevil, Otiorhynchus sulcatus, though the type of formulation and the specific isolate used must be selected carefully.     

Nucleotide excision repair and photoreactivation in the entomopathogenic fungi Beauveria bassiana, Beauveria brongniartii, Beauveria nivea, Metarhizium anisopliae, Paecilomyces farinosus and Verticillium lecanii

AIMS: To compare the DNA repair capabilities of the entomopathogenic fungus (EPF) bassiana to the EPF Beauveria brongniartii, Beauveria nivea, Metarhizium anisopliae, Paecilomyces farinosus, Verticillium lecanii, and the fungi Aspergillus niger and Neurospora crassa. METHODS AND RESULTS: Germination of B. bassiana conidiospores following ultraviolet (UV) irradiation was used to show that nucleotide excision repair and photoreactivation decrease the post-UV germination delay. These two modes of repair were characterized and compared between the aforementioned EPF, A. niger and N. crassa using a physiological assay where per cent survival post-UV irradiation was scored as colony forming units. CONCLUSIONS: The results showed B. bassiana and M. anisopliae are the most UV-tolerant EPF. The DNA repair capabilities indicated that EPF do not have all DNA repair options available to fungi, such as A. niger and N. crassa. SIGNIFICANCE AND IMPACT OF THE STUDY: A key factor detrimental to the survival of EPF in agro-ecosystems is UV light from solar radiation. The EPF literature pertaining to UV irradiation is varied with respect to methodology, UV source, and dose, which prevented comparisons. Here we have characterized the fungi by a standard method and established the repair capabilities of EPF under optimal conditions.     

Effects of Beauveria bassiana and Metarhizium anisopliae on mortality, fecundity and egg fertility of Tetranychus evansi

Abstract:  The susceptibility of various developmental stages of Tetranychus evansi Baker & Pritchard (eggs, larvae, protonymphs, deutonymphs and adults) to the entomopathogenic fungi Metarhizium anisopliae (Metschnikoff) Sorokin and Beauveria bassiana (Balsamo) Vuillemin was evaluated under laboratory conditions. Three concentrations (3.0 × 10⁶, 1.0 × 10⁷ and 1.0 × 10⁸ conidia/ml) of both fungi were used for each stage. The effect of fungal infection on fecundity and egg fertility was also investigated using both fungal species. Deutonymphs that survived the infection and developed into adult females were allowed to oviposit. Adults and deutonymphs were more susceptible to fungal infection than larval and protonymphal stages at all the concentrations. Nevertheless, the concentration level influenced the mortality of the different mite stages. Eggs were also susceptible to fungal infection and mortality was dose-dependent. Fungus-treated female mites laid fewer eggs than the controls but there was no significant difference in egg hatchability between the treatments.     

Pathogenicity of the entomogenous fungi Metarhizium anisopliae and Beauveria bassiana against crucifer pests and the honey bee

The susceptibility of the cabbage stem flea beetle (Psylliodes chrysocephala; Coleoptera: Chrysomelidae) to different isolates of the entomogenous, hyphomycete fungi Beauveria bassiana and Metarhizium anisopliae, all from heterologous hosts, was investigated. The most pathogenic isolates were of M. anisopliae (V208, V233, V234, V242, V245, and V248). Studies were conducted to determine the virulence of V208 and V245, and their specificity for three other crucifer pests, Phaedon cochleariae, Myzus persicae and Lipaphis erysimi and a beneficial insect, Apis mellifera. Flea beetle mortality rates increased with dose; the estimated LC₅₀ ^S of V208 and V245 at 14 days post‐inoculation were c. 10⁶ conidia ml^‐1. The respective estimated LT₅₀ values of V208 and V245 for P. chrysocephala at 1 × 10⁷ conidia ml^‐1 were 10.0 and 9.3 days, and at 1 × 10¹⁰ conidia ml LT₅₀s were 3.8 and 4.2 days. Both V208 and V245 were pathogenic for P. cochleariaeM. persicae and L. erysimi. Myzus persicae and L. erysimi died within 4 days of inoculation. Bees were less susceptible to infection at low doses, but at 1 × 10¹⁰ conidia ml^‐1almost all bees had died by day 5 when the estimated LT₅₀s for V208 and V245 were 4.4 and 8.5 days respectively.     

航天诱变对昆虫病原真菌的生物学效应

8个昆虫病原真菌菌株经过航天搭载后,有5个菌株的孢子在航天搭载后全部死亡,菌株M2029和MR8的存活率仅约10-7,菌株M189的存活率可达到20%以上。分离得到航天诱变的单菌落菌株,检测表明航天诱变使昆虫病原真菌的形态、生长速度、产孢量、致病力等多项生物学特性发生了不同程度的变异,变异率高,变异趋向有正负双向性。测定结果表明航天诱变菌株HM189-68s、HM189-32c、HM189-127和HM189-17的生长特性和致病力优于原始菌株。航天诱变为选育生物防治优良菌株提供了新途径。     

Use of entomopathogenic fungi combined with biorational insecticides to control Dichroplus maculipennis (Orthoptera: Acrididae: Melanoplinae) under semi-field conditions

Grasshoppers are among the invertebrate herbivores that cause most economic losses in grasslands throughout Argentina’s Pampas and parts of Patagonia. Chemical insecticides remain the sole option for grasshopper control in this area, despite being of significant environmental concern. Our aim was to evaluate the efficacy of combinations between three entomopathogenic fungi strains (Beauveria bassiana LPSc 1067 and LPSc1082), and Metarhizium anisopliae (LPSc 907), two biorational insecticides (luphenuron and methoxyfenozide), and a new synthetic chemical pesticide (rynaxypyr) in the control of the pest grasshopper Dichroplus maculipennis under field cage conditions. Fungal strains used were adjusted to 1 × 10⁸, 1 × 10⁶ and 1 × 10⁴ conidia/ml. Insecticides were tested at three concentrations: the average concentration recommended for application in the field (100%), 50% of that level and finally 25%. Combinations of the insecticides with B. bassiana (LPSc 1067, LPSc 1082) and M. anisopliae (LPSc 907) caused higher mortality to D. maculipennis nymphs than any of the individual agents used alone. The three insecticides tested did not negatively affect the isolates of the two species of entomopathogenic fungi employed.     

Influence of whey permeate and millet as substrates on thermotolerance of Beauveria bassiana and Metarhizium anisopliae conidia during storage

Conidia from Beauveria bassiana and Metarhizium anisopliae, produced in quarter-strength Sabouraud dextrose agar amended with yeast extract, whey permeate, and millet agar, were exposed to 45°C for 90 min prior to and at 30 d post-storage at 25°C. B. bassiana produced in whey permeate or millet had better thermotolerance than the other treatments after the storage.     

Pathogenicity of Beauveria bassiana,Metarhizium anisopliae and Serratia marcescens to the banana weevil Cosmopolites sordidus

Two exotic fungal isolates, one of Beauveria bassiana (268–86) and another of Metarhizium anisopliae (100–82), three local isolates of B. bassiana (isolates I, II, III) and one of the entomogenous bacteria Serratia marcescens, were tested for pathogenicity against the banana weevil Cosmopolites sordidus. All four isolates of B. bassiana and the one of M. anisopliae were found to be pathogenic to third—instar larvae of C. sordidus, causing mortalities of 98–100% by 9 days post—exposure to dry fungal spores. M. anisopliae was the least pathogenic to larvae with LT₅₀ of 4.2 days, compared to 3.5, 3.3, 3.6 and 4.0 respectively for isolates I, II, III and 268–86. B. bassiana was also pathogenic to adult C. sordidus, causing mortalities varying from 63–97% by 35 days post—exposure depending on isolate. As for larvae M. anisopliae exhibited low pathogenicity for the adult C. sordidus. In general, all the fungi tested were less pathogenic to adult weevils (LT₅₀ = 17.5; 12.5; 8.0 and 22.0 days) for isolates I, II, III and 268–86 respectively, while isolate 100–82 failed to kill 50% of adults even by 35 days post—exposure. Incubation of dead weevils in a moist environment led to development of surface mycelia starting from intersegmental junctions. Histopathology revealed extensive destruction of internal organs by hyphae which invaded most of the organs. The LT₅₀ for S. marcescens against C. sordidus larvae was 2.8 days. However, the bacterium did not kill adult C. sordidus even at 10 times the concentration applied on larvae.     

Persistence and virulence of promising entomopathogenic fungal isolates for use in citrus orchards in South Africa

Thaumatotibia leucotreta (Meyrick) (1912) (Lepidoptera: Tortricidae) is a key economic pest of citrus in South Africa. Although a variety of control options are available, none currently registered target the soil-dwelling life stages of the pest. Three fungal isolates, two Metarhizium anisopliae sensu lato Metschnikoff (Sorokin) isolates (G 11 3 L6 and FCM Ar 23 B3) and one Beauveria bassiana sensu lato Balsamo (Vuillemin) isolate (G Ar 17 B3) have been identified as showing the greatest potential against T. leucotreta soil-dwelling life stages. Since environmental persistence is an important factor in the success of entomopathogenic fungi against soil pests, the isolates, along with two commercial isolates (B. bassiana Eco-Bb® and M. anisopliae ICIPE 69), were subjected to a six-month field persistence trial. Every month after burial of the fungal isolates, inside net bags, in orchard soil, the number of colony forming units (CFUs) per gram of soil was measured and the viability of the conidia was assessed using bioassays. All isolates investigated were capable of persisting in the soil for the duration of the trial period and in addition, were capable of initiating infection in T. leucotreta late fifth instar larvae and subsequent pupae. However, with the exception of the commercial isolates which showed some correlation, no correlation was found between the number of CFUs recorded and percentage T. leucotreta mycosis. Persistence of entomopathogenic fungi in the field is affected by a series of factors, and although the effect of these factors was not measured, the data have provided preliminary evidence towards the persistence capability of these candidate biopesticides against T. leucotreta.     

白僵菌、绿僵菌对雷公藤丽长角巢蛾的侵染力测定

经室内侵染力测定,球孢白僵菌Beauveria bassiana和金龟子绿僵菌Metarhizium anisopliae皆能侵染雷公藤丽长角巢蛾Xyrosaria lichneuta幼虫,不同菌株的侵染力存在显著差异。3株白僵菌中,Bb27菌株毒力最强,菌液为1×10^5～1×10^9个·mL^-1五个不同浓度梯度的孢子悬浮液时,害虫15d校正死亡率在58．3％-100％之间;LT50分别为9．36d,6．26d,3．89d、2．54d．2．10d;LC50为4．87×10^4个·ML^-1。3株绿僵菌中,MaZPTP01菌株毒力最强,5种孢子浓度15d校正死亡率分别为52．4％、69．0％、76．2、92．9％、95．2％;LT50分别为11．47d、6．89d、4．36d、2．68d、2．13d;LC50为7．39×10^4个·mL^-1。Bb27和MaZFIT01菌株可应用于林间作进一步防治试验。     

Effect of combination treatment with entomopathogenic fungi Beauveria bassiana and Nomuraea rileyi (Hypocreales) on Spodoptera litura (Lepidoptera: Noctuidaeae)

In biocontrol of insect pests, efficacy of treatment with multiple pathogens has not been frequently investigated but may have potential for effective management. The possible advantage of a combination treatment with two entomopathogenic fungi - Beauveria bassiana and Nomuraea rileyi - was assessed in laboratory bioassays on second instar Spodoptera litura. From among the fungal isolates of an epizootic population, two isolates of each fungus differing in virulence to S. litura were chosen, one highly virulent and the other with low virulence. The bioassays were carried out at either a continuous temperature of 25±1°C or at a temperature cycle of 32±2°C 8 h/21±2°C 16 h to mimic the field temperatures during the epizootic. Treatments with the two fungi were done both simultaneously and sequentially. In combination treatments at 25±1°C, in all isolate combinations, a majority of the larvae showed N. rileyi induced mycosis; the percentage mortality and speed of kill of insects in these treatments was similar to the N. rileyi isolate used in the combination treatments. At the temperature cycle of 32±2°C 8 h/21±2°C 16 h, in all combination treatments, all the dead insects exhibited B. bassiana mycosis; the mortality pattern was similar to the B. bassiana isolate used in the combination treatments. The adverse effect of high temperature on virulence of N. rileyi was however, not evident in in vitro growth assays. Combination treatment with both fungi did not have a synergistic effect on insect mortality.     

Effect of entomopathogenic fungi upon adults of Stomoxys calcitrans and Musca domestica (Diptera: Muscidae)

The pathogenicity and virulence of 10 isolates of entomopathogenic fungi from the soil of lodging pens of dairy production units in Aguascalientes, Mexico, on adults of Stomoxys calcitrans and Musca domestica were determined. All isolates were pathogenic when exposed by aspersion to a concentration of 1×10⁸ conidia/ml, causing between 20.3 and 91.7% mortality in S. calcitrans and between 31 and 91.7% in M. domestica at 7 days post-exposure; in S. calcitrans isolates of Beauveria bassiana (Bb114) and Metarhizium anisopliae (Ma135), sensu lato, were the most noteworthy as mortality reached above 90% with an LC₅₀ of 3.5×10⁵ conidia/ml for Bb114, while for Ma135 reached 1.6×10⁴ conidia/ml. In M. domestica Ma134 and Ma135 showed mortality above 90% with an LC₅₀ of 4.3×10⁴ and 1.4×10⁵ conidia/ml, respectively.     

Bioassay method for assessing the virulence of Beauveria bassiana against tarnished plant bug, Lygus lineolaris (Hem., Miridae)

A standard bioassay method for assessing the pathogenicity of Beauveria bassiana (Balsamo) Vuillemin (GHA strain) against second instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hem., Miridae) was developed. Several types of inoculation methods, assay containers and incubation times were tested. Our goal was to minimize control mortality and maximize treatment mortality. Five inoculation methods (immersing broccoli florets or bean pods, spraying broccoli florets or bean pods, and immersing insects) and four types of plastic containers (114‐, 171‐, 228‐ and 455‐ml) were tested. Immersing insects directly in a fungal suspension was the most effective inoculation method, which resulted in a treatment mortality of 70–81.3% at a concentration of 1 × 10⁷ conidia/ml. The 114‐ml plastic container was the most suitable assay container when 10 tarnished plant bug nymphs were treated together, resulting in a control mortality of only 6% 12 days after treatment. Within the first 6 days after treatment, 71.1% of the insects were killed, compared with a total mortality of 81.3% after 12 days. Nymphs infected with the test fungus changed colour from green to black. Mycelial outgrowth and sporulation on the cadavers demonstrated that most nymphs died of fungal infection. A total of 61.1 and 80.5% of the cadavers showed signs of mycelial outgrowth 9 days after death among those that were surface sterilized and those that were not, respectively.     

Efficacy and molecular studies of a Lebanese isolate of Beauveria for control of Thaumetopoea wilkinsoni (Lepidoptera: Thaumetopoeidae)

A Lebanese isolate of a Beauveria species originally isolated from a hymenopterous insect pest was found to be very effective against the pine processionary moth Thaumetopoea wilkinsoni. In laboratory bioassays, high mortality rates of the five larval stages were attained with spore concentrations of about 5000–50,000 spores/larva. The efficacy of kill on the first three larval stages was similar to the growth regulator diflubenzuron, but it was significantly higher on the fourth and fifth larval stages. Sequences of the DNA lyase gene and the EF-1α gene were used for molecular characterisation of this Beauveria isolate. The DNA lyase gene showed more polymorphism than the previously reported ITS region and EF-1α gene. This constitutes the first report on the possibility of using the DNA lyase gene as a molecular tool in fungal taxonomy.     

Effect of nutrition on growth and virulence of the entomopathogenic fungus Beauveria bassiana

Three isolates of the entomopathogen Beauveria bassiana along with one strain of Metarhizium anisopliae were cultured on seven media with different carbon/nitrogen (C/N) ratios. The effect of nutrition on virulence of the isolates was evaluated via measurement of colony growth, spore yield, germination speed, conidial C/N ratio and Pr1 (a serine protease) activity. 'Osmotic stress' medium produced the lowest colony growth with low numbers of conidia in all isolates. However, these conidia showed a high germination rate and virulence. However, conidial Pr1 activity was low in some isolates. In most but not in all cases conidia from 1% yeast extract, 2% peptone and low (10 : 1) C/N medium had higher Pr1 activity compared with conidia from other media. However, in some instances we could not conclude that there was a relationship among germination rate, conidial Pr1 activity and virulence. C/N ratio of conidia was statistically different among various media and fungal isolates. Conidia with lower C/N ratio generally produced lower LT(50) (lowest median lethal time) values (more virulent). Insect-passaged conidia from different media had lower C/N ratio compared with similar conidia from artificial cultures. Therefore, they should be more virulent than in vitro produced conidia. As germination rate, conidial Pr1 activity and C/N ratio are independent of host, it seems that host-related determinants such as insect cuticle and physiology and environmental conditions may influence host susceptibility and therefore fungal isolate virulence towards host insects.