Enhanced mortality of Bemisia tabaci nymphs by Isaria javanica combined with sublethal doses of chemical insecticides

The aim of this current study was to evaluate the mortality of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) nymphs by the combination between the entomopathogenic fungus Isaria javanica (Friedrichs & Bally) Samson & Hywel‐Jones and synthetic chemical insecticides. The bioefficacy of I. javanica and the insect growth regulators named spiromesifen and buprofezin was tested alone and in combination against B. tabaci nymphs under screenhouse conditions. The in vitro compatibility between these two control agents was previously assessed under laboratory conditions. The sublethal concentration (LC₂₅) of these insecticides towards second‐instar nymphs was determined and then mixed with the fungal treatments to investigate the type of interaction. All I. javanica isolates at 5 × 10⁷ conidia/ml inflicted nymphal mortality by up to 62.4%. The insecticides did not influence the germination and mycelial growth of the selected I. javanica isolate CG1282. In general, the insecticide–fungus combinations increased nymphal mortalities in comparison with their single counterparts. Combinations with the fungus and buprofezin or spiromesifen augmented nymphal mortality by 10% and 24%, respectively, in comparison with the fungus alone. Additive interaction was found with the combination of the I. javanica CG1282 at 1 × 10⁶ conidia/ml and spiromesifen at 1.56 ppm, and additive and synergistic interactions were achieved with the fungus at 5 × 10⁶ conidia/ml and spiromesifen at 3.12, 6.25 and 12.5 ppm. Additive interactions were also observed from mixtures of fungus at 5 × 10⁶ and buprofezin at 3.12 and 6.25 ppm. Only combinations of I. javanica with spiromesifen reduced to some extent the mycosis on dead nymphs. The joint action of I. javanica with sublethal doses of these insecticides may be a promising tool to assist in the integrated management system for B. tabaci.     

Susceptibility of the tobacco whitefly,Bemisia tabaci (Hemiptera: Aleyrodidae) biotype Q to entomopathogenic fungi

Recently, the Q biotype of tobacco whitefly has been recognized as the most hazardous strain of Bemisia tabaci worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the susceptibility of this biotype of B. tabaci to 21 isolates of Beauveria bassiana, three isolates of Isaria fumosorosea, one isolate of I. cateni, three isolates of Lecanicillium lecanii, one isolate of L. attenuatum, and one isolate of Aschersonia aleyrodis. These isolates were evaluated on pruned eggplant seedlings, at a concentration of 10⁸ conidia/mL (deposited at 6000±586 conidia mm^?2). The mortality based on mycosis varied from 18 to 97% after 6 days. Isaria fumosorosea isolate Pf04, B. bassiana isolates Bb06, Bb12, and L. lecanii L14 were found the most effective. Furthermore, five isolates were chosen for concentration–mortality response assays and compared to B. bassiana GHA as a standard. The numbers of nymphs infected by fungi were correlated with the spore concentration. L. lecanii L14 and I. fumosorosea Pf04 had the shortest LT₅₀ at 3.5 and 3.3 days at 6000±586 conidia mm^?2. Mortality declined and LT₅₀ values were longer as the concentration of conidia was reduced. The LD₅₀ values were calculated as 87, 147, 191, 263, and 269 conidia mm^?2 for isolates L14, GHA, AS1260, Bb13, and Pf04, respectively. These results indicated that the Q biotype of sweetpotato whitefly was susceptible to the five isolates of entomopathogenic fungi and these isolates have potential to be developed as microbial pesticides for whitefly control.     

Effect of host plant on susceptibility of whitefly Bemisia tabaci (Homoptera: Aleyrodidae) to the entomopathogenic fungus Beauveria bassiana (Ascomycota: Hypocreales)

We determined host plant effect on susceptibility of whitefly Bemisia tabaci to the entomopathogenic fungus Beauveria bassiana under controlled conditions. Insects were reared on cucumber, eggplant, tomato or cabbage. Fungal suspensions of 1×10⁴, 10⁵, 10⁶, 10⁷ and 10⁸ conidia/mL were applied on second-instar nymphs. Nymphal survival significantly differed among different host plant species on which the nymphs were reared. Ten days after inoculation with 1×10⁸ conidia/mL, percent survival was 4.2±0.7, 9.6±0.4, 13.4±0.8, and 24.3±0.9% on cucumber, eggplant, tomato and cabbage, respectively. Average survival times of nymphs were also significantly influenced by host plant species. After inoculation with 1×10⁸ conidia/mL, survival times were 4.8±0.15, 6.0±0.11, 5.7±0.13, and 6.2±0.08 days for nymphs reared on cucumber, eggplant, tomato, and cabbage, respectively. Virulence also differed depending on host plant species; 10 days after inoculation, LC₅₀ values were 4.6×10⁴, 1.6×10⁵, 4.2×10⁵ and 2.1×10⁶ conidia/mL on cucumber, eggplant, tomato and cabbage, respectively. Nymphs on cucumber showed highest susceptibility.     

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.      

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

LABORATORY SUSCEPTIBILITY OF PLUTELLA XYLOSTELLA TO METARHIZIUM ANISOPLIAE AND NOMURAEA RILEYI*

Abstract Five isolates of Metarhizium anisopliae and one isolate of Nomuraea rileyi were bioassayed against larvae of Plutella xylostella. Larvae were treated by exposing cabbage leaf discs previously immersed in conidial suspension, and were then incubated at 25^oC and observed daily. One of M. anisopliae isolates, F11248, originally from Mastotermes darwiniensis was found being most virulent against P. xylostella. The LC₅₀ was estimated as 2.03 times 10⁴conidia/ml with 9 d mortality data, and the LT₅₀ was 4.97 d at concentration of 10⁷conidia/ml. Isolate F163 (N. rileyi) showed virulence to P. xylostella in both tests, but no cadavers sporulated.     

The potential of Beauveria bassiana inoculum formulated into a polymeric matrix for a microbial control of spruce bark beetle

Two local strains of Beauveria bassiana originally isolated from naturally infected spruce bark beetles in Slovakia were tested for their virulence to Ips typographus (IT) and for their compatibility with a polymeric matrix composed of low-molecular polyethylene. Conidia could be homogenously immobilized in the low-molecular polyethylene matrix with no adverse effect on their viability and infectivity. At constant temperature (25°C), viability of immobilized conidial decreased only by 1–2% after 7 or 14 days when compared with non-formulated conidia. In field conditions, viability of conidia formulated in the matrix was even significantly higher than non-formulated conidia 35 days after their application in traps. Conidia incorporated into the polymeric matrix were infective to IT adults in laboratory bioassays. Mean values of LC₅₀ for native conidia (0.72–2.05?×?10⁶ conidia?ml^?1) and conidia immobilized in the polymeric matrix (0.64–1.03?×?10⁵ conidia?mm^?2) demonstrated high virulence. The efficacy of the local strains was significantly higher than that of B. bassiana strains from mycoinsecticides (Boverol^®, Botanigard^® ES and Naturalis-L^®). Results showed potential of this polymeric material for its use in microbial control of IT when mixed with conidia of B. bassiana.     

Cellular and humoral immune defenses of Oxya japonica (Orthoptera: Acrididae) to entomopathogenic fungi Metarhizium anisopliae

One Indonesian isolate of the fungus Metarhizium anisopliae, named Majalengka strain, was evaluated not only for its virulence but also for the immune response of rice grasshopper Oxya japonica (Orthoptera: Acrididae) as a target organism. Five aqueous suspensions with different conidia concentrations in logarithmic series were prepared. The fungus showed high virulence as it caused 100% mortality at low conidia concentration (1.5 × 10² conidia/mL). Remarkable changes in the cellular and humoral responses were also observed when adult grasshoppers were infected with the fungus. The number of hemocytes decreased significantly within 12 h after infection. In addition, the total number of granulocytes increased rapidly in the first 12 h then gradually decreased 24 and 48 h after infections, while the number of coagulocytes fluctuated over time. The infection influenced the humoral response by increasing the phenoloxidase activity.     

安徽虫瘟霉的黍米培养及其对桃蚜的侵染力

安徽虫瘟霉Zoophthora anhuiensis (Li) Humber是较难人工培养的蚜虫专化性病原真菌。将灭菌并适度熟化的黍米Panicum miliaceum L.作为固体基质与挑碎的安徽虫瘟霉平板菌落混合,在20℃和12L∶12D的温光条件下静止固体培养,获得了产孢潜能大、杀蚜活性强的米粒培养物。培养7天的黍米的产孢量达13.0×10⁴个孢子/粒,产孢持续时间长达6天。用此黍米培养物弹射的孢子对桃蚜Myzus persicae (Sulzer) 若蚜进行7.9～134.9个孢子/mm²共9个剂量的孢子浴接种,所获数据很好拟合时间 剂量 死亡率模型。接种后第5～7天各天的LC₅₀依次为59.8, 39.5和33.5个孢子/mm²,LC₉₀依次为354,234和198个孢子/mm²。在57.7～134.9个孢子/mm²的接种剂量范围内,致死中时LT₅₀从5.1天下降到4.3天。由此表明,安徽虫温霉的黍米培养不仅简单易行,而且菌种的产孢和侵染生物学特性在培养物中被充分体现,每颗米粒如同自然罹病而死的蚜尸,值得进一步研究开发和利用。     

Different host plant species modifies the susceptibility of Bactericera cockerelli to the entomopathogenic fungus Beauveria bassiana

Bactericera cockerelli (Sulc.) is a serious pest of solanaceous crops and a vector of the plant pathogen Candidatus Liberibacter psyllaurous. Entomopathogenic fungi are the most important biological control alternatives for this pest. Host plant species, however, can modify the outcomes of insect–pathogen interactions. We conducted laboratory experiments to quantify the virulence of two isolates of the entomopathogenic fungus Beauveria bassiana (Bals. [Vuill.]), BB40 and BB42, against third instar B. cockerelli nymphs maintained on chilli pepper plants. Owing to the lack of difference in virulence against B. cockerelli nymphs on chilli pepper between the two B. bassiana isolates, only BB42 was used to: compare virulence against nymphs maintained on either chilli pepper, potato or tomato; and in vivo conidia production from nymphs maintained on different host plants. Virulence of the two B. bassiana isolates against B. cockerelli nymphs was similar. Bactericera cockerelli nymphs maintained on tomato were more susceptible to B. bassiana than nymphs maintained on potato or chilli peppers. Infected nymphs maintained on chilli peppers produced the greatest number of conidia followed by infected nymphs maintained on tomato and potato. Host plant affected the susceptibility of B. cockerelli to B. bassiana isolate BB42 and subsequent conidia production. The implications of our results for microbial control of B. cockerelli by B. bassiana are discussed.     

Identification of a bacterium isolated from the diseased brown planthopper and determination of its insecticidal activity

The brown planthopper, Nilaparvata lugens, is one of the most harmful insect pests of rice crops in Asian countries. To find an effective biological control agent against this pest, we investigated the bacterial flora of field N. lugens collected from Jiangsu Province, China, in 2012 and tested its insecticidal activity. A novel bacterium strain, S-JS1, was isolated from N. lugens nymphs and adults and showed a high level of insecticidal activity. Based on its phenotypic, physiological and biochemical properties, and its 16S rRNA gene phylogeny, the isolate was assigned to Serratia marcescens; the name S. marcescens S-JS1 is proposed. The pathogenicity of S-JS1 against the third-instar nymphs, and the macropterous and the brachypterous adults of N. lugens were compared. The median lethal concentration (LC₅₀) values of S-JS1 against the brachypterous adult were the lowest (LC₅₀, 1.53?×?10⁸ colony forming units (cfu)/ml), followed against the macropterous adult (LC₅₀, 1.65?×?10⁹?cfu/ml) and third-instar nymphs (LC₅₀, 1.86?×?10⁹?cfu/ml) at 5 days post-infection. The median lethal time values of 8?×?10⁸?cfu/ml S. marcescens S-JS1 against the brachypterous adult, macropterous adult, and third-instar nymph were 4.5, 5.5, and 5.7 days, respectively. These results indicate that the S-JS1 isolate appears to be a promising S. marcescens strain with strong biocontrol potential against N. lugens.     

Host plant influences pathogenicity of Beauveria bassiana to Bemisia tabaci and its sporulation on cadavers

Laboratory bioassays were conducted to determine host plant effect on pathogenicity of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. (Ascomycota: Hypocreales) to the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Fourth instar B. tabaci reared on cucumber, tomato, melon, green pepper, potato, eggplant, marrow, cabbage, bean or cotton, were treated with 1 × 10⁷ conidia/ml B. bassiana EABb 93/14-Tp isolate. Mortality caused by B. bassiana significantly increased with time and it was significantly affected by the host plant on which the nymphs were reared. Mean mortality of nymphs 8 days after inoculation ranged between 52.3±7.3 for nymphs reared on cotton and 91.8±5.8 for nymphs reared on cucumber. Average survival times of nymphs treated with the fungal suspensions were also significantly influenced by the host plant, with a mean of 4.7±0.1 days for nymphs reared on cucumber, 6.6±0.2 days for cotton and 6.9±0.1 days for green pepper. The production of newly formed conidia was also affected by host plant and varied from 111000±8600 conidia/cadaver for nymphs reared on cotton to 597000±28000 conidia/cadaver for those reared on melon.     

Selection of promising fungal biological control agent of the western flower thrips Frankliniella occidentalis (Pergande)

Aims: Larval stages of Frankliniella occidentalis are known to be refractory to fungal infection compared with the adult stage. The objective of this study was to identify promising fungal isolate(s) for the control of larval stages of F. occidentalis. Methods and Results: Ten isolates of Metarhizium anisopliae and eight of Beauveria bassiana were screened for virulence against second‐instar larvae of F. occidentalis. Conidial production and genetic polymorphism were also investigated. Metarhizium anisopliae isolates ICIPE 7, ICIPE 20, ICIPE 69 and ICIPE 665 had the shortest LT₅₀ values of 8·0–8·9 days. ICIPE 69, ICIPE 7 and ICIPE 20 had the lowest LC₅₀ values of 1·1 × 10⁷, 2·0 × 10⁷ and 3·0 × 10⁷ conidia ml^?1, respectively. Metarhizium anisopliae isolate ICIPE 69 produced significantly more conidia than M. anisopliae isolates ICIPE 7 and ICIPE 20. Internally transcribed spacers sequences alignment showed differences in nucleotides composition, which can partly explain differences in virulence. Conclusion: These results coupled with the previous ones on virulence and field efficacy against other species of thrips make M. anisopliae isolate ICIPE 69 a good candidate. Significance and Impact of the Study: Metarhizium anisopliae isolate ICIPE 69 can be suggested for development as fungus‐based biopesticide for thrips management.     

Insecticidal activity of entomopathogenic fungi (Hypocreales) for potato psyllid,Bactericera cockerelli (Hemiptera: Triozidae): Development of bioassay techniques,effect of fungal species and stage of the psyllid

The potato psyllid, Bactericera cockerelli (?ulc), is a pest of potato, tomato, and some other solanaceous vegetables and has also been incriminated in the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum, resulting in a serious disease known as ‘zebra chip’. Although there are several reports of fungal pathogens in psyllids, there are none from B. cockerelli, nor have any fungi been evaluated against it. Five isolates of fungi, one Beauveria bassiana, two Metarhizium anisopliae and two Isaria fumosorosea, were bioassayed against B. cockerelli on potato leaves under ideal conditions for the fungi. All applications were made with a Potter spray tower. With the exception of concentration-effect studies, all other applications were made using 10⁷ conidia/mL in a 2-mL aqueous suspension. All isolates except B. bassiana, produced 95–99% mortality, corrected for control mortality, in adults 2–3 days after application of conidia and 91–99% in nymphs 4 days after application. The corrected mortalities for adults and nymphs treated with B. bassiana were 53 and 78%, respectively, 4 days after application. I. fumosorosea Pfr 97 produced 95% corrected mortality in both first and late third instar nymphs. M. anisopliae (F 52) produced 96% corrected mortality in first and third instar nymphs. Pfr 97 and F 52 were evaluated for insecticidal activity against third instar B. cockerelli using 10⁵, 10⁶, and 10⁷ conidia per mL. Mortality produced by I. fumosorosea Pfr 97 ranged from 83 to 97% and that of M. anisopliae F 52 was 88 to 95% at these concentrations.     

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.     

Evaluation of the pathogenicity of Aschersonia aleyrodis on Bemisia tabaci in the laboratory and greenhouse

The entomopathogenic fungus Aschersonia aleyrodis (Webber) is a promising fungal species against whiteflies. In this work, the pathogenicity of A. aleyrodis isolate Aa005 against MEAM1 Bemisia tabaci (Gennadius) was evaluated under laboratory and greenhouse conditions. The bioassay results indicated that the percentage of larval mortalities was concentration and age dependent. A. aleyrodis showed high pathogenicity against second and third instars and pupae with LC₅₀ values of 7.93?×?10⁶, 1.08?×?10⁷, and 1.56?×?10^7?conidia?mL^?1, respectively. The median lethal time (LT₅₀) was lower (4.60 days) for second instars and was the highest (6.17 days) for pupae when inoculated with a concentration of 1?×?10^7?conidia?mL^?1. Weekly sampling of immatures showed that the per cent mortality caused by A. aleyrodis at a conidial concentration of 1?×?10^7?conidia?mL^?1 was 71.21% in small nymphs, 69.31% in large nymphs and 53.36% in pupae. The dispersion index (DI) and Lloyd’s Index of Patchiness (LIP) values indicated that the infected immatures had a tendency to aggregate. The study demonstrated that A. aleyrodis isolate A005 is an effective biocontrol agent for B. tabaci control under laboratory and greenhouse conditions.     

Susceptibility of Larvae of Galleria mellonella to Infection by Aspergillus fumigatus is Dependent upon Stage of Conidial Germination

The ability of conidia of the human pathogenic fungus Aspergillus fumigatus to kill larvae of the insect Galleria mellonella was investigated. Conidia at different stages of the germination process displayed variations in their virulence as measured using the Galleria infection model. Non-germinating (‘resting’) conidia were avirulent except when an inoculation density of 1 × 10⁷ conidia per insect was used. Conidia that had been induced to commence the germination process by pre-culturing in growth medium for 3 h were capable of killing larvae at densities of 1 × 10⁶ and 1 × 10⁷ per insect. An inoculation density of 1 × 10⁵ conidia per insect remained avirulent. Conidia in the outgrowth phase of germination (characterised as the formation of a germ tube) were the most virulent and were capable of killing 100% of larvae after 5 or 24 h when 1 × 10⁷ or 1 × 10⁶ conidia, that had been allowed to germinate for 24 h, were used. Examination of the response of insect haemocytes to conidia at different stages of the germination process established that haemocytes could engulf non-germinating conidia and those in the early stages of the germination process but that conidia, which had reached the outgrowth stages of germination were not phagocytosed. The results presented here indicate that haemocytes of G. mellonella are capable of phagocytosing A. fumigatus conidia less than 3.0 μm in diameter but that conidia greater than this are too large to be engulfed. The virulence of A. fumigatus in G. mellonella larvae can be ascertained within 60–90 h if infection densities of 1 × 10⁶ or 1 × 10⁷ activated conidia (pre-incubated for 2–3 h) per insect are employed.     

Susceptibility of Ixodes scapularis (Acari: Ixodidae) to Metarhizium brunneum F52 (Hypocreales: Clavicipitaceae) using three exposure assays in the laboratory

An emulsifiable concentrate (EC) and granular (G) formulation of the entomopathogenic fungus, Metarhizium brunneum strain F52 (formerly Metarhizium anisopliae strain F52) were tested against unfed adults and nymphs of Ixodes scapularis Say in the laboratory. Three exposure methods; dip, surface contact, and direct spray application, and three exposure time intervals (3, 30, and 300 min) were used to evaluate the EC formulation. Application rates ranged from 2.6 x 10(2) to 2.6 x 10(8) conidia/cm2. The surface treatment was used for granular formulation with concentrations ranging from 2.3 x 10(5) to 2.3 x 10(7) conidia/cm2 for same three exposure times. Both the EC and G formulations of this fungus were highly pathogenic against I. scapularis adults and nymphs. Logistic regression analysis found formulation, spore concentration, time of exposure, and observation period were significant or highly significant factors influencing tick mortality. For adult I. scapularis, the spray application with the EC formulation of M. brunneum F52 resulted in a lower LC50 (5.9 x 10(4) conidia/cm2) at 30 min than surface exposure to the EC (LC50 = 1.3 x 10(6) conidia/cm2) or G formulation (LC50 = 8.1 x 10(5) conidia/cm2). At higher concentrations, fungal activity was evident in adult I. scapularis held at 5 degrees C suggesting the fungus may provide control in the cooler fall season. While the observed pathogenicity of a fungus against ticks can be dependent upon the bioassay assessment, we found nymphs and adults of I. scapularis to be highly susceptible to M. brunneum F52, regardless of the exposure method used.     

Time-dose-mortality modelling and virulence indices for six strains of Verticillium lecanii against sweetpotato whitefly, Bemisia tabaci (Gennadius)

Bioassays of six strains of Verticillium lecanii (Zimmermann) Viégas were conducted with the sweet potato whitefly, Bemisia tabaci. For inoculation, batches of third‐instar whitefly nymphs on sweetpotato seedlings were immersed in conidial suspensions of five dosages at concentrations from 10³ to 10⁷ conidia/ml. Each dosage was used to inoculate 150–250 nymphs. The nymphs were maintained at 25°C, under 95% RH, photo phase of 16 : 8 (L : D) and observed daily for mortality. The resulting data were analysed over 8 days by a complementary log‐log (CLL) time‐dose‐mortality model, based on the Hosmer–Lemeshow test, analysing the time‐dose trends for five concentrations of six strains of V. lecanii simultaneously. The parameters from the model were used to estimate the virulence indices (the values of LC₅₀) of six strains against B. tabaci. Based on the time‐dose‐mortality relationships fitted and the virulence indices, the virulence of the six strains of V. lecanii for B. tabai was compared. Results indicated that the strain Vl6063 imported from Canada and the domestic strains V3450 and Vp28 derived from B. tabaci and a scale insect, respectively, were more virulent than the others with LC₅₀ values of 2.57 × 10⁵, 6.03 × 10⁵ and 6.03 × 10⁵, respectively.     

Assessment of oral virulence against Spodoptera litura,acquired by a previously non-pathogenic Metarhizium anisopliae isolate,following integration of a midgut-specific insecticidal toxin

All entomopathogenic fungi infect insects by direct penetration through the cuticle rather than per os through the gut. Genetic transformation can confer fungi with per os virulence. However, unless the recipient isolate is nonpathogenic to the target insect, mortality caused by a transgenic isolate cannot be attributed solely to oral virulence due to the potential for some simultaneous cuticular infection. Here, a Metarhizium anisopliae wild-type isolate (MaWT) nonpathogenic to Spodoptera litura was genetically engineered to provide a transformed isolate (MaVipT31) expressing the insect midgut-specific toxin Vip3Aa1. Toxin expression was confirmed in MaVipT31 hyphae and conidia using Western blotting. Mortality, leaf consumption and body weight of S. litura larvae (instars I–IV) exposed to a range of concentrations of MaWT conidia were not significantly different to controls although the number of conidia ingested by surviving larvae during the bioassay ranged from 2.3 × 10⁵ (instar I) to 8.1 × 10⁶ (instar IV). In contrast, consumption of MaVipT31 conidia caused high mortalities, reduced leaf consumption rates and decreased body weights in all instars evaluated, demonstrating that oral virulence had been acquired by MaVipT31. Larval mortalities were much more dependent on the number of MaVipT31 conidia ingested than the duration of time spent feeding on conidia-treated leaves (r²: 0.83–0.94 for instars I–IV). LC₅₀ and LT₅₀ trends for MaVipT31 estimated by time-concentration-mortality modeling analyses differed greatly amongst the instars. For 50% kill to be achieved, instar I larvae required 3, 4 and 5 days feeding on the leaves bearing 103, 28 and 8 conidia/mm² respectively; instar IV larvae required 6, 7 and 8 days feeding on leaves bearing 1760, 730 and 410 conidia/mm² respectively. Our results provide a deeper insight into the high oral virulence acquired by an engineered isolate and highlight its great potential for biological control.