Influences of host volatiles on feeding behaviour of the Japanese pine sawyer, Monochamus alternatus

Abstract: Choice and no‐choice feeding assays on the twigs of three host species demonstrated the following feeding preference sequence by Monochamus alternatus: Pinus massoniana > Cedrus deodara > Pinus thunbergii. There were significant differences in the concentrations of α‐pinene, camphene, d ‐limonene, β‐phellandrene, longifolene and β‐caryophyllene in volatiles emitted by twigs among the three species. We tested the effects of six monoterpenes (α‐pinene, β‐pinene, 3‐carene, myrcene, limonene and β‐caryophyllene) added to an artificial diet consisting of bark from P. thunbergii on consumption rates by M. alternatus. The addition of α‐pinene at all four concentrations 0.4, 1.2, 3.6 and 10.8 μl/ml resulted in increases of a twofold greater consumption rate than the control at a concentration of 3.6 μl/ml. Limonene inhibited diet consumption at concentrations >0.4 μl/ml. The concentration of α‐pinene in volatiles emitted by twigs was significantly higher for P. massoniana than for P. thunbergii, whereas the reverse was true for limonene. There were no differences for any of the other host components, suggesting that α‐pinene and limonene may play an important role in the adult's selection and acceptance of suitable and unsuitable feed host. Mixed compounds promoted the consumption of artificial diet at a concentration of 0.4 μl/ml, whereas consumption was inhibited at a concentration of 10.8 μl/ml. There were significant linear correlations (β‐pinene: r² = 0.930, P < 0.05; myrcene: r² = 0.933, P < 0.05) between the amount of diets consumed and diet concentrations of β‐pinene and myrcene. In conclusion, host volatile terpenes may stimulate or repel M. alternatus depending on terpene concentrations they encounter during initial feeding and then possibly inhibit further feeding activity once concentrations increase to threshold levels.     

Cold tolerance and cold hardening strategy of the Japanese pine sawyer Monochamus alternatus (Coleoptera: Cerambycidae)

The Japanese pine sawyer, Monochamus alternatus , is an important pine forest pest and vector transmitting the pine wilt nematode that causes pine wilt disease. Low temperatures in autumn, winter and spring often differentially affect mortality of M. alternatus larvae. In this paper, we mainly compared the differences of mortality and cold hardening of larvae from different seasons, based on supercooling point (SCP) and cumulative probability of individuals freezing (CPIF). The cold hardening of the larvae from autumn, winter and spring seasons were largely different. Correlations between mortality and CPIF of autumn and spring larvae were highest on day 1/4, and gradually decreased with prolonged exposure duration. This beetle's death mainly resulted from freezing in short exposure duration. However, the correlation between mortality and CPIF of winter larvae increased gradually with the prolonged exposure duration. Death did not mainly result from freezing in long exposure duration. Autumn larvae are more susceptible and adaptable than winter and spring larvae. Winter larvae have a slight freeze-tolerance trend. Our research showed that M. alternatus came into complex cold-hardening strategies under natural selection. Freeze avoidance is the primary strategy; with prolonged exposure duration to above SCP or < 0 °C, chill tolerance is more important; this is followed by freeze tolerance during harsh winters.     

Oviposition schedules,survivorship curves,and mortality factors within trees of two cerambycid beetles (Coleoptera: Cerambycidae), the Japanese pine sawyer,Monochamus alternatus hope,and sugi bark borer,Semanotus japonicus lacordaire

Summary Oviposition schedules under laboratory conditions, survivorship curves, and mortality factors within trees of two cerambycid beetles, Japanese pine sawyer (JPS),Monochamus alternatus Hope, and sugi bark borer (SBB),Semanotus japonicus Lacordaire, were investigated. Average longevities of reared adults of JPS were 38.1 days for males and 42.3 days for females; those of SBB were 15.7 days for males and 23.8 days for females. It was confirmed that the JPS must feed on the pine branches for full maturation after emergence, but SBB need not; they are able to lay eggs soon after emergence. The average fecundities of JPS and SBB were 32.9 and 90.5, respectively. Thus, the JPS lay fewer eggs for a long time with continuous maturation feeding, whereas the SBB lay more eggs for short time without maturation feeding. Average survivorship curves of JPS within dead pine trees in 8 pine forests were theDeevey's B type, showing a constant mortality through the pre-imaginal stages in the trees. On the other hand those of SBB in 4 cedar stands approached theDeevey's type, suggesting that the high mortality occurred at an early stage in the trees. Average mortalities of JPS between the appearance of oviposition scars and adult emergence in 8 forests ranged from 62.3% to 95.2%. Intraspecific ompetition of JPS resulting from overcrowding in dead pine trees appeared to be main mortality factor. For SBB, as most larvae were killed by resin flow in living trees, this appeared to be the main mortality factor for this species.     

Oviposition deterrence associated with larval frass of the Japanese pine sawyer,Monochamus alternatus (Coleoptera: Cerambycidae)

To determine the effect of larval frass of Monochamus alternatus on oviposition preference of the female adults, three tests were performed in the laboratory. Individual females were provided with a frass-coated, Pinus densiflora bolt and an untreated bolt simultaneously and were allowed to oviposit for 24 h. They deposited a significantly smaller number of eggs on frass-coated bolts. The females supplied with frass-coated bolts deposited no eggs on them, indicating that the larval frass deterred the females from oviposition. When individual females were provided simultaneously with a pine bolt applied with a methanol extract of larval frass and another bolt applied with methanol alone, they deposited a significantly smaller number of eggs on frass extract-applied bolts for 24 h. These results suggest the presence of a putative oviposition deterrent in larval frass of M. alternatus. Each test was replicated 19-20 times using different females.     

Role of host volatiles in mate location by the Japanese pine sawyer, Monochamus alternatus hope (Coleoptera: Cerambycidae)

We evaluated the responses of male and female Monochamus alternatus Hope (Coleoptera: Cerambycidae) to various terpenes commonly associated with host trees. Electroantennogram (EAG) tests were conducted with 12 plant volatile compounds and ethanol. Antennae of both sexes were highly sensitive to (R)-(+)-alpha-pinene, (+)-3-carene, (-)-beta-pinene, and terpinolene. Both sexes of M. alternatus were attracted by traps baited with (+)-alpha-pinene, (-)-beta-pinene, (+)-3-carene, or terpinolene. Our results support the first of the three-stage hypothesis posed by Ginzel and Hanks that suggests that location of stressed trees by cerambycids involves three stages: (1) both sexes locate larval hosts by using plant volatiles as kairomones; (2) males produces sex pheromones to attract females after both sexes land on the larval hosts; (3) males and female recognize each other by contract pheromones in their epicuticular wax layer. Males and females showed differences in their EAG responses to several compounds, including (R)-(+)-alpha-pinene, (-)-beta-pinene, myrcene, (+)-3-carene, (R)-(+)-limonene, terpinolene, and trans-caryophyllene. In all cases, males exhibited greater sensitivity than females. In laboratory assays, male M. alternatus showed strong preference for 1% (+)-alpha-pinene and 1% (-)-beta-pinene over other compounds. In field assays, traps baited with (+)-alpha-pinene, (-)-beta-pinene, (+)-3-carene, or terpinolene caught more beetles than control traps. We found strong male bias in beetle catches in baited traps and those captured on the stem of stressed trees despite a strong female bias in emerging beetles in 2004. We hypothesize that male M. alternatus are more responsive than females to plant volatiles and that males have more capacity than females in finding mating locations.     

Application of harmonic radar to analyze dispersal behavior of the Japanese pine sawyer beetle,Monochamus alternatus (Coleoptera: Cerambycidae)

Monochamus alternatus (Hope) is a severe wood‐boring pest in coniferous forests and a major vector of pine wilt disease in East Asia. Harmonic radar is a powerful tool for studying the dispersal behavior of insects and it could be applied to control pine wilt disease. In this study, we validated the application of harmonic radar for analyzing the dispersal behavior of M. alternatus beetles in a field environment. We determined the wing capacities of the beetles and the effects of electronic tagging and marking on their movement, flight ability, survivorship, and food consumption in the laboratory to confirm the suitability of this technique. The detection rate and recovery rate for beetles were analyzed separately using radar on caged pine stands and in the field environment. The results showed that the minimum wing capacity of the Japanese pine sawyer was 24.9 mg, which was seven times the weight of the electronic tag (3.5 mg). Marking and tagging the beetles had no significant adverse effects on their movement, flight capacity, food consumption, and survivorship. The detection rate using the radar system and recovery rate based on visual observations of the beetles in caged pines were both 95.6%. However, in the field environment, the detection and recovery rates were only 55.6% and 37.8% after one week, respectively, and 33.3% and 7.8% after two weeks. Harmonic radar is a promising technique for studying the dispersal behavior of the Japanese pine sawyer, but its performance is not satisfactory and major improvements are required for both the radar system and electronic tags.     

Feeding and oviposition preference of the Sakhalin pine sawyer Monochamus saltuarius (Coleoptera: Cerambycidae) for various tree species

Monochamus saltuarius was recently identified as an important vector of Bursaphelenchus xylophilus, the cause of pine wilt disease. In this study, M. saltuarius adults preferred different species of tree based on the longevity, ingested amount and wounded area that were determined for 90 d. Among eight tree species, the longevity of feeding by M. saltuarius was not significantly different except when feeding in Sciadopitys verticillata. However, the highest ingested amounts with the largest wounded areas were associated with Pinus koraiensis. The highest oviposition preference was in P. densiflora for the first 20 d. However, after 40 d, the oviposition highest preference was noted in P. koraiensis. For oviposition activities, M. saltuarius preferred the tree species in the following order: P. densiflora > P. koraiensis > P. rigida > Larix leptolepis. Although many oviposition scars were observed, M. saltuarius laid fewer eggs in P. rigida than other species of pine, which was related to the bark thickness. The oviposition frequencies were highest with bark 2.0–2.5 mm thick and with tree diameters of 110–130 mm. The current study increased understanding of the ecology of M. saltuarius by identifying specific tree preferences.     

Smoke,pheromone and kairomone olfactory receptor neurons in males and females of the pine sawyer Monochamus galloprovincialis (Olivier) (Coleoptera: Cerambycidae)

The response of antennal olfactory receptor neurons (ORNs) of Monochamus galloprovincialis to several odourants was tested using single sensillum electrophysiology. Behaviourally active pheromone, and kairomone (host and sympatric bark beetle pheromone) odours were tested alongside smoke compounds released by burnt wood that are potentially attractive to the insect. The antennae bore several types of sensilla. Two plate areas in the proximal and distal ends of each antennal segment were covered with basiconic sensilla that responded to the odour stimuli. Sensilla basiconica contained one or two cells of different spike amplitude. The 32 male and 38 female ORNs tested responded with excitations or inhibitions to the different plant odours. In general the response of male and female receptors was very similar so they were pooled to perform a cluster analysis on ORN responses. Six ORNs were clearly specialised for pheromone reception. Responses to kairomone and smoke odours were less specific than those of pheromone, but a group of 9 cells was clearly excited by smoke compounds (mainly eugenol and 4-methyl 2-methoxyphenol), a group of 8 cells was very responsive to α-pinene, β-pinene and cis-verbenol, and a group of 14 cells responded to a wider range of compounds. The rest of the cells (47%) were either non-responsive or slightly inhibited by smoke compounds. Dose–response curves were obtained for several compounds. Different compounds induced significantly different latencies and these appeared to be unrelated to their boiling point.     

Pathogenicity of entomopathogenic fungus,Metarhizium anisopliae MET-GRA4 isolate on dengue vectors,Aedes albopictus and Aedes aegypti mosquito larvae (Diptera: Culicidae)

Considering the rapid transmission of the dengue virus, substantial efforts need to be conducted to ward-off the epidemics of dengue viruses. The control effort is depending on chemical insecticides and had aroused undesirable conflicts of insecticide resistance. Here, we study the entomopathogenic fungus, Metarhizium anisopliae as a promising new biological control agent for vector control. The pathogenicity effects of Metarhizium anisopliae against field and laboratory strains of Aedes albopictus and Aedes aegypti larvae were tested using the larvicidal bioassay technique. The results demonstrate that the treatments using M. anisopliae isolate MET-GRA4 were highly effective and able to kill 100% of both Ae. albopictus and Ae. aegypti mosquito larvae at a conidia concentration of 1 × 10?/ml within 7 days of the treatment period. The fungus displayed high larvicidal activity against laboratory and field strain of Ae. aegypti larvae with LC₅₀ values (9.6 × 10³/ml, 1.3 × 10³/ml) and LC₉₅ values (1.2 × 10?/ml, 5.5 × 10⁵/ml) respectively. For Ae. albopictus, LC₅₀ values for laboratory and field strains were (1.7 × 10⁴/ml, 2.7 × 10⁴/ml) and the LC₉₅ values were (2.1 × 10?/ml, 7.0 × 10⁵/ml) respectively. Interestingly, the susceptibility of field strain towards M. anisopliae was higher as compared to the laboratory strain Aedes larvae. In which, the causative agents of all the dead larvae were verified by the virulence of M. anisopliae and caused morphological deformities on larval body. The findings from this study identify this isolate could be an effective potential biocontrol agent for vector mosquitoes in Malaysia.     

A novel,easy method for estimating pheromone trap attraction range: application to the pine sawyer beetle Monochamus galloprovincialis

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Lethal effects of Gliocladium virens,Beauveria bassiana and Metarhizium anisopliae on the malaria vector Anopheles albimanus (Diptera: Culicidae)

Control of Anopheles albimanus, the main vector of malaria on the coast of the State of Chiapas, is based mainly on application of chemical insecticides, which has resulted in resistance to most registered insecticides. Strategies for biological control may provide sustainable alternatives. We report on the lethal effects of a native isolate of Gliocladium virens on An. albimanus larvae and adults, compared to that of strains of Beauveria bassiana and Metarhizium anisopliae. Conidial suspensions of G. virens, B. bassiana and M. anisopliae cultured on Sabouraud agar were tested in bioassays with An. albimanus larvae and adults. Mosquito larvae were more susceptible to all fungi, compared to adults. On early and late instar larvae, M. anisopliae showed the most pathogenic effect (LC₅₀ of 1.4×10⁵ conidia/mL in early instars; 1.1×10⁵ conidia/mL in late instars), followed by G. virens (LC₅₀ of 3.3×10⁵ conidia/mL in early instars and 3.5×10⁶ conidia/mL in late instars). Metarhizium anisopliae sensu lato and the native G. virens could be considered good choices for An. albimanus control in southern Mexico.     

Resporulation of Metarhizium anisopliae granules on soil and mortality of Tenebrio molitor: Implications for wireworm management in sweetpotato

In Australia, sweetpotato (Ipomoea batatas L.) is vulnerable to root feeding insect pests such as wireworms (e.g., Agrypnus spp.). The number of registered insecticides to control these insect pests is limited and often pest pressure, for example by wireworms, is severe close to harvest, further limiting what insecticides can be applied. Incorporating biological control agents such as entomopathogenic fungi (e.g., Metarhizium anisopliae) into integrated pest management programmes may be feasible in sweetpotato. M. anisopliae has been shown to be effective in controlling more than 200 insects and it is able to reside and grow in the rhizosphere and rhizoplane, suggesting that M. anisopliae could be a promising candidate against soil insect pests. In the study presented here, M. anisopliae was formulated into calcium alginate granules fortified with nutrients. The resporulation of the fungal granules was tested on four different soil types in the laboratory. The biocontrol efficacy of the resulting fungal growth was also examined using larval mealworms, Tenebrio molitor as a model insect in the laboratory and the glasshouse. Our results indicated that sterilised soil favoured optimal fungal resporulation, although different soil types did not have a significant effect on fungal resporulation. The resulting fungal resporulation and growth on sterilised soil caused high mortality (up to 76%) of larval mealworms in the glasshouse, whereas the fungal granules applied to non-sterile soil demonstrated poor resporulation that led to low mortality (13%) of larval mealworms. The result of this study indicates that the manipulation of microbial populations in field soil is required to enhance the fungal growth and potential insect control against wireworms in the field.     

Characterization of Beauveria bassiana and Metarhizium anisopliae isolates for management of tarnished plant bug,Lygus lineolaris (Hemiptera: Miridae)

Selected morphological and physiological characteristics of four Beauveria bassiana (Balsamo) Vuillemin isolates and one Metarhizium anisopliae (Metschnikoff) Sorokin isolate, which are highly pathogenic to Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), were determined. There were significant differences in conidial size, viability, spore production, speed of germination, relative hyphal growth, and temperature sensitivity. Spore viability after incubation for 24h at 20 degrees C ranged from 91.4 to 98.6% for the five isolates tested. Spore production on quarter-strength Sabouraud dextrose agar plus 0.25% (w/v) yeast extract after 10 days incubation at 20 degrees C ranged from 1.6x10(6) to 15.5x10(6)conidia/cm(2). One B. bassiana isolate (ARSEF 1394) produced significantly more conidia than the others. Spore germination was temperature-dependant for both B. bassiana and M. anisopliae. The time required for 50% germination (TG(50)) ranged from 25.0 to 30.9, 14.0 to 16.6, and 14.8 to 18.0h at 15, 22, and 28 degrees C, respectively. Only the M. anisopliae isolate (ARSEF 3540) had significant spore germination at 35 degrees C with a TG(50) of 11.8h. A destructive sampling method was used to measure the relative hyphal growth rate among isolates. Exposure to high temperature (40-50 degrees C) for 10min had a negative effect on conidial viability. The importance of these characteristics in selecting fungal isolates for management of L. lineolaris is discussed.     

Virulence, horizontal transmission, and sublethal reproductive effects of Metarhizium anisopliae (Anamorphic fungi) on the German cockroach (Blattodea: Blattellidae)

Virulence of Metarhizium anisopliae (Metschnikoff) Sorokin strain EAMa 01/121-Su against the German Cockroach, Blatella germanica (L.), was determined using four concentrations ranging from 4.2 x 10(6) to 4.2 x 10(9) spores per milliliter. The LD50 value was 1.4 x 10(7) spores per milliliter (56,000 spores per cockroach) and LT50 values were 14.8 days and 5.3 days for 4.2 x 10(8) and 4.2 x 10(9) spores per milliliter, respectively. An experiment was conducted to evaluate whether a fungal transmission could exist among infected and healthy cockroaches. Percentage mortality at a ratio of 1:10 of infected to unexposed cockroaches was 87.5% and LT50 was 12.2 days, which indicated the potential of this strain to be horizontally transmitted and to rapidly spread the infection in the insect population. The effect of a sublethal dose (ca. LD60) of M. anisopliae EAMa 01/121-Su strain, applied topically on German cockroaches, was studied by reciprocal crossing. Othecal production, oothecal hatchability, and nymphal production declined upon exposure to M. anisopliae EAMa 01/121-Su strain. The mean number of oothecae laid by female was progressively and significantly reduced by fungal treatment from second oviposition period onwards. Oothecal hatch of fungally challenged females was reduced by 46-49%, oothecal viability by 48-85%, and nymphal production by 22-35%. Only treated females showed an effect on oothecal production, oothecal hatch, and nymphal production, although oothecal hatch was also governed by treated males at a higher significance level. Our results on virulence and horizontal transmission of fungal conidia of M. anisopliae EAMa 01/121-Su strain and its sublethal reproductive effects on German cockroach females are discussed in terms of its potential to decrease the pest status of B. germanica in the short and long terms.     

室内条件下莱氏猛叩甲对松墨天牛和黄粉虫的捕食能力

【目的】本研究旨在明确莱氏猛叩甲Tetrigus lewisi老熟幼虫分别对黄粉虫Tenebrio molitor和松墨天牛Monochamus alternatus末龄幼虫和蛹的捕食量的差异,评估莱氏猛叩甲的捕食能力,为松墨天牛M.alteratus的有效治理找到潜在的高效生物防治因子。【方法】于2014年11月在浙江富阳昌东村马鞍山砍下松墨天牛虫害木,通过林间野外调查,明确松墨天牛和莱氏猛叩甲的发生情况;将野外采集到的莱氏猛叩甲幼虫带回室内,分别饲喂黄粉虫和松墨天牛的末龄幼虫和蛹,每天记录捕食量,确定莱氏猛叩甲的捕食能力。【结果】野外调查中,我们在松墨天牛幼虫蛀食的坑道中发现了莱氏猛叩甲幼虫和松墨天牛幼虫被取食后的残体;在采集到1 094头松墨天牛幼虫的危害木内,共采得莱氏猛叩甲末龄幼虫36头,约为松墨天牛幼虫数量的3%。室内捕食实验表明,5 d内莱氏猛叩甲捕食黄粉虫幼虫总量为3.2头,平均每天捕食约0.6头,而捕食松墨天牛幼虫总量为8.0头,平均每天捕食约1.6头,莱氏猛叩甲对松墨天牛的总捕食量显著高于对黄粉虫幼虫的总捕食量(P0.0001);4 d内莱氏猛叩甲捕食黄粉虫蛹总量为5.6头,平均每天捕食约1.4头,而捕食松墨天牛蛹总量为7.8头,平均每天捕食约2.0头,莱氏猛叩甲对松墨天牛蛹的捕食量显著高于黄粉虫蛹(P=0.028)。回归分析还表明,莱氏猛叩甲幼虫的捕食量不受叩甲个体大小的影响。【结论】莱氏猛叩甲普遍分布在松墨天牛发生区,同时发现松墨天牛被取食后的残体,表明莱氏猛叩甲是松墨天牛的捕食性天敌。莱氏猛叩甲均能捕食黄粉虫和松墨天牛的幼虫和蛹;相比于捕食黄粉虫,莱氏猛叩甲对松墨天牛的幼虫和蛹具有更好的捕食效率,具有很好的生物防治前景,是未来松墨天牛治理潜在高效生物防治因子。     

Persistence of Metarhizium anisopliae incorporated into soilless potting media for control of the black vine weevil, Otiorhynchus sulcatus in container-grown ornamentals

The objective of this study was to determine the persistence of Metarhizium anisopliae (F52), measured as infectivity against black vine weevil larvae, in a soilless potting medium at six wholesale nursery locations across the Willamette Valley, Oregon. A granule formulation (0.30 and 0.60 kg/m(3)) was incorporated into media at planting and fungal persistence determined over two growing seasons. The fungus persisted in the potting media over the duration of the experiment with 50-60% of the larvae exposed to treated media becoming infected at the end of the experiment. The percentage of infected larvae gradually declined from > or = 90% on week 3 to 40-60% by week 19. Larval infection rebounded over the fall and winter months of 2004 to 75-80% followed again by a slow decline over the course of the second growing season.     

Evaluation of potential entomopathogenic fungus,Beauveria bassiana,for controlling the coffee berry borer Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae) in Taiwan

Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), a coffee berry borer (CBB), is a serious pest of coffee fruits and causes economic losses in the coffee industry. With the concerns of chemical control, an alternative eco-friendly management strategy such as microbial control has emerged as the times require. Herein, a total of thirty-three fungi were isolated from CBB cadavers. Two entomopathogenic fungal isolates, NCHU-271 and NCHU-272, which showed fast insect-killing activity, were subjected to molecular identification and named “Beauveria bassiana-NCHU-271” (Bb-NCHU-271) and “Beauveria bassiana -NCHU-272” (Bb-NCHU-272). Moreover, a virulence test of Bb-NCHU-271 and Bb-NCHU-272 against CBBs was also performed. Both Bb-NCHU-271 and Bb-NCHU-272 caused 100% mortality at 8 days post inoculation (d.p.i.) and a LT₅₀ of Bb-NCHU-271 by spraying 10⁸ conidia/ml was less than Bb-NCHU-272. Therefore, histological evidence of Bb-NCHU-271 infected CBBs was provided to proven the infection process of B. bassiana in CBBs. Furthermore, an infestation test was performed to evaluate the differences in CBB control efficacy between the treatments of “Borers exposed to EPF first” or “Berries exposed to EPF first”. The results revealed that the “Borers exposed to EPF first” of Bb-NCHU-272 caused significantly lower survival rates and higher mycosis rates than those of Bb-NCHU-271. Only slight control effects of Bb-NCHU-271 and Bb-NCHU-272 were also observed in the “Berries exposed to EPF” treatment. In conclusion, Bb-NCHU-271 and −272 showed controlling ability to the CBBs either before or after CBB infestation into coffee berries, while application of EPFs before the CBBs bored into coffee fruits could enhance the control effect.     

The influence of temperature on the susceptibility of vine weevil, Otiorhynchus sulcatus (Fabricius) (Coleoptera: Curculionidae), larvae to Metarhizium anisopliae (Deuteromycotina: Hyphomycetes)

Mortality of Otiorhynchus sulcatus (Fabricius) larvae at 10^oC, 15^oC, 20^oC and 25^oC following treatment with 10⁷conidia ml“¹suspensions of six Metarhizium anisopliae (Metschnikoff) Sorokin isolates was temperature dependent. In all cases, the LT₅₀s were inversely related to temperature, but the nature of this response varied between isolates. Strain 101-82 was the most virulent isolate at 25^oC with an LT₅₀ of 3.7 days, but it was the least virulent isolate at 15^oC and it failed to kill any O. sulcatus larvae at 10^oC. In contrast, strain 159-83 had the lowest virulence at both 20^oC and 25^oC, whereas it was the most virulent isolate at 10^oC with an LT₅₀ of 20.0 days. The mortality rates followed a similar pattern and were positively related to temperature in all cases with the exception of strain 159-83 at 25^oC. Mycosis development was examined on larvae treated with strain 275-86 and significant differences were obtained between all four temperatures. Sporulation commenced after approximately 2.75 days at 25^oC, but took nearly 11 days at 10^oC. The infection rates also varied between temperatures; sporulation occurred on 98% of the treated larvae at 25^oC, but only on 93%, 87% and 49% of the larvae at 20^oC, 15^oC and 10^oC, respectively. The results of these bioassays demonstrate that temperature has a significant effect on the virulence of M. anisopliae. The differences between fungal strains also emphasises the importance of selecting isolates for specific situations on the basis of their temperature profiles.     

Food consumption by Chilo partellus (Lepidoptera: Pyralidae) larvae infected with Beauveria bassiana and Metarhizium anisopliae and effects of feeding natural versus artificial diets on mortality and mycosis

Second and third instar Chilo partellus larvae were infected with Beauveria bassiana and Metarhizium anisopliae (both at 1x10(8)conidia/ml) and daily consumption of maize leaves was measured. Infection by the fungi was associated with reduced mean daily food consumption. Reduction in food consumption became evident 3-4 days after treatment with the fungi for second instar larvae and 4-5 days for third instar larvae. Four conidial concentrations, 1x10(5), 1x10(6), 1x10(7), and 1x10(8)conidia/ml, were tested against second instar larvae. Food consumption dropped by 70-85% when the second instar larvae were treated with the fungi at 1x10(8)conidia/ml. Reduction in food consumption by C. partellus larvae infected with B. bassiana and M. anisopliae may offset the slow speed of kill of the fungi. The effect of artificial versus natural diets on mortality and mycoses of second instar larvae treated with the fungi at 1x10(8)conidia/ml was determined. Larvae provided with artificial diet suffered little mortality and mycoses than larvae provided with maize leaves. The LT(50) was longer for larvae provided with artificial diet.