Evaluation of entomopathogenic fungus Metarhizium anisopliae against soil-dwelling stages of cabbage maggot (Diptera: Anthomyiidae) in glasshouse and field experiments and effect of fungicides on fungal activity

The effect of two isolates of the entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (389.93 and 392.93) on root-feeding stages of cabbage root fly, Delia radicum (L.), was studied under glasshouse and field conditions. In glasshouse studies, the effect of drenching a suspension of conidia (concentration 1 x 10(8) m(-1), 40 ml per plant, applied on four occasions) onto the base of cabbage plants infested with D. radicum eggs was compared with mixing conidial suspension into compost modules (concentration 1 x 10(8) ml(-1), 25 ml per plant) used to raise seedlings. Drench application reduced the mean number of larvae and pupae recovered per plant by up to 90%, but the compost module treatment had no statistically significant effect. Both application methods reduced the emergence of adult flies from pupae by up to 92%. Most conidia applied as a drench application remained in the top 10-cm layer of compost. Applications of the fungicides iprodione and tebuconazole, which are used routinely on brassica crops, were compatible with using M. anisopliae 389.93 against D. radicum under glasshouse conditions, even though these fungicides were inhibitory to fungal growth on SDA medium. In a field experiment, drench applications of M. anisopliae 389.93 to the base of cauliflower plants at concentrations of 1 x 10(6) to 1 x 10(8) conidia ml(-1) did not control D. radicum populations, although up to 30% of larval cadavers recovered supported sporulating mycelium. Drench applications often exhibited considerable lateral movement on the soil surface before penetrating the ground, which may have reduced the amount of inoculum in contact with D. radicum larvae.     

Mortality of Delia floralis, Galleria mellonella and Mamestra brassicae treated with insect pathogenic hyphomycetous fungi

Abstract:  The susceptibility of Delia floralis eggs, neonates and larvae and the susceptibility of Galleria mellonella and Mamestra brassicae larvae to seven different Norwegian isolates of the insect pathogenic, hyphomycetous fungi Tolypocladium cylindrosporum , Metarhizium anisopliae and Beauveria bassiana , were investigated. Metarhizium anisopliae isolate ARSEF 5520 was highly virulent to G. mellonella larvae and caused 100% mortality when tested at a concentration of 3.6 × 10⁶ conidia/ml. The same M. anisopliae isolate was not virulent to D. floralis larvae. Isolates of T.cylindrosporum , were equally virulent to G. mellonella and D. floralis causing up to 36.0% mortality of larvae. It is suspected, however, that the use of grated rutabaga as a food source in the D. floralis bioassay reduced the fungal virulence of both M. anisopliae and T. cylindrosporum to D. floralis . Among three T. cylindrosporum isolates tested at a concentration of 1.0 × 10⁷ conidia/ml against eggs of D. floralis none of them reduced the hatching percentage. One isolate, ARSEF 5525 did, however, significantly reduce the longevity of neonates. Beauveria bassiana isolates ARSEF 5510 and ARSEF 5370 tested at a concentration of 1.0 × 10⁷ conidia/ml resulted in M. brassicae mortality levels of 70.0 and 55.0%, respectively. The B. bassiana isolate ARSEF 5557, however, was not virulent to M. brassicae . Among the three isolates tested against M. brassicae the two virulent isolates produced a red pigment, probably oosporein, when cultured in Sabouraud dextrose agar.     

Potential of some Metarhizium anisopliae isolates for control of Culex quinquefasciatus (Dipt., Culicidae)

Abstract: The potential of some isolates of Metarhizium anisopliae and Beauveria bassiana for use in the integrated management of Culex quinquefasciatus was evaluated. Metarhizium anisopliae isolate 1037 was selected in initial bioassays with a 50% lethal concentration (LC₅₀) of 1.97 × 10⁴ conidia/ml. This fungus caused higher mosquito larva mortality when applied as a conidial suspension to the surface of the water than as dry conidia, with a time to 50% lethal (LT₅₀) of 1 day compared with 3.6 days for the dry conidial application. However, results with UV- and heat-inactivated conidia did not confirm a possible role of fungal toxins in causing mortality when ingested by C. quinquefasciatus larvae. Metarhizium anisopliae did not remain active for as long as the bacterium Bacillus sphaericus isolate 2362. At 24 h after application, the fungus-induced mortality on mosquito larvae was significantly lower than the mortality caused by the bacterium. By the second day, almost no activity by the fungus was observed. Results suggest that M. anisopliae isolate 1037 has potential for use in mosquito control programmes.     

Selection of a highly virulent fungal isolate, Metarhizium anisopliae CLO 53, for controlling Hoplia philanthus

The June beetle, Hoplia philanthus Füessly (Coleoptera: Scarabaeidae), has become a widespread and destructive insect pest of lawns, sport turf, pastures, and horticultural crops in Belgium. The virulence of 34 entomopathogenic fungal isolates from the genera Metarhizium, Beauveria, and Paecilomyces to third-instar H. philanthus was tested in bioassays by dipping larvae in 10(7)conidia/ml suspensions. Two isolates of Metarhizium anisopliae (CLO 53 and CLO 54) caused maximally 90% mortality 10 weeks post-inoculation while other isolates only caused mortalities between 10 and 62%. The virulence of M. anisopliae CLO 53 was further tested by exposing H. philanthus larvae to conidial serial concentrations of 10(4)-10(9)conidia/g sandy soil for up to 11 weeks at 15, 20 or 25 degrees C. Mortality was dependant on the fungal concentration, exposure time, and temperature. Eleven weeks after inoculation, the LC50 values for this isolate ranged from 1.3 to 4.0 x 10(6), 1.0 to 3.2 x 10(5), and 2.5 x 10(4) to 10(5)conidia/g soil at 15, 20, and 25 degrees C, respectively. The LT50 values for this isolate ranged from 3.5 to 21.7, 2.4 to 18.7, and 2.9 to 16.1 weeks at concentrations of 10(9) and 10(4)conidia/g soil at 15, 20, and 25 degrees C, respectively. In glasshouse pot experiment with perennial ryegrass (Lolium perenne L.), the isolate CLO 53 caused mortalities of 50 and 88% of H. philanthus larvae 10 weeks after application of 10(4) and 10(6)conidia/cm(2) soil surface, respectively. The present results suggest that the Belgian isolate CLO 53 has excellent potential for biological control of H. philanthus.     

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.     

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.     

Effect of temperature and poultry litter in Beauveria bassiana (bals.) Vuill. and Metarhizium anisopliae (Metsch) virulence against the lesser mealworm Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

This study was carried out to evaluate the influence of temperature and poultry litter on germination vegetative growth virulence and conidial production of Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) isolates on larvae and adults of the lesser mealworm (Alphitobius diaperinus) (Panzer). The vegetative growth and conidial production were evaluated on culture media. Virulence was studied submerging larvae and adults in a conidial suspension (1 x 10(8) conidia/ml). All the experiments were carried out in growth chamber (26 degrees C and 32 degrees C and 14h photophase). Fungus-killed insects were daily collected and used for microscopic conidial counts. The poultry litter effect was evaluated by submerging the insects in a fungal suspension (10(8) conidia/ml) and then transferring them to cups containing poultry litter (new and used). B. bassiana isolates were more sensitive than M. anisopliae to high temperature because conidia viability, vegetative growth and virulence were negatively affected (P < 0.05). The conidial production was higher to B. bassiana in 26 degrees C (7 to 11 x 10(8) conidia/larval cadaver and 8 x 10(8) conidia/adult cadaver) (P < 0,05). Larval stage was about 10 times more sensitive to M. anisopliae at 26 degrees C than adults stage. Regarding B. bassiana, differences on sensitivity between larval stages and adults were not observed at this temperature. However, at 32 degrees C, larval stage was more sensitive for CB116 and UEL50 isolates. Mortality was higher when larvae and adults (15.7 and 66.7% respectively) were treated by B. bassiana and maintained on new poultry litter at 26 degrees C) (P < 0.05).     

Evaluation of Metarhizium anisopliae (Deuteromycota: Hyphomycetes) for control of broad mite Polyphagotarsonemus latus (Acari: Tarsonemidae) in mulberry

A study on 12 entomopathogenic fungi for controlling broad mite (Polyphagotarsonemus latus (Banks)) in mulberry found that Metarhizium anisopliae CKM-048 was the most virulent strain in controlling both larvae and adult broad mites at the concentration of 2 x 10(8) conidia/ml. There was no ovicidal effect when tested with broad mite eggs. Median lethal concentrations (LC(50)) of M. anisopliae in killing larvae and adults were 8.7 x 10(6) and 1.3 x 10(7 )conidia/ml, respectively. Median lethal times (LT(50)) of larvae and adults were 2.4 and 3.8 days, respectively, at the concentration of 2 x 10(8) conidia/ml. The fungus was found to produce protease and chitinase. Scanning electron microscope (SEM) studies were done to monitor the infection steps of the fungus on broad mites. A greenhouse test on mulberry trees revealed that M. anisopliae could reduce the broad mite population within 4 days after treatment. However, after 7 days, its efficacy was decreased significantly.     

Susceptibility of Agrilus planipennis (Coleoptera: Buprestidae) to Beauveria bassiana and Metarhizium anisopliae

The susceptibility of Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) to selected strains of the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin was evaluated through bioassays with direct immersion or foliar exposure under laboratory conditions. Results showed that A. planipennis adults were susceptible to B. bassiana and M. anisoplae. Significant time-mortality response was found for each isolates. Isolate B. bassiana GHA killed A. planipennis adults at a faster rate compared with other isolates tested, with the lowest average time-to-death values. The LC50 values estimated under direct immersion method ranged from 1.7 x 10(5) to 1.9 x 10(7), 3.5 x 10(4) to 5.3 x 10(5), and 4.1 x 10(3) to 2.9 x 10(5) conidia/ml for B. basissiana and from 3.2 x 10(6) to 1.1 x 10(7), 4.5 x 10(3) to 4.5 x 10(5), and 1.4 x 10(2) to 1.2 x 10(5) conidia/ml for M. anisopliae at 4, 5, and 6 d after treatment, respectively. By days 5 and 6, B. bassiana GHA outperformed all other isolates tested except ARSEF 7234, followed by ARSEF 7152, 6393, and 7180. Significant concentration-mortality response was also observed for two B. bassiana GHA formulations, BotaniGard ES and Mycotrol O, and M. anisopliae F52 when insects were treated through foliar exposure. The LC50 values ranged from 114.5 to 309.6, 18.4 to 797.3, and 345.3 to 362.0 conidia/cm2 for BotaniGard, Mycotrol, and M. anisopliae F52, respectively. Based on the results of these bioassays, the efficacy of both B. bassiana GHA formulations and M. anisopliae F52 were similar against adult A. planipennis. The potential use of entomopathogenic fungi for management of A. planipennis in North America is discussed.     

Susceptibility of different developmental stages of large pine weevil Hylobius abietis (Coleoptera: Curculionidae) to entomopathogenic fungi and effect of fungal infection to adult weevils by formulation and application methods

The large pine weevil, Hylobius abietis, is a major pest in European conifer forests causing millions of Euros of damage annually. Larvae develop in the stumps of recently felled trees; the emerging adults feed on the bark of seedlings and may kill them. This study investigated the susceptibility of different developmental stages of H. abietis to commercial and commercially viable isolates of entomopathogenic fungi, Metarhizium and Beauveria. All the developmental stages of H. abietis can be killed by Metarhizium robertsii, Metarhizium brunneum, and Beauveria bassiana. The most virulent isolate of M. robertsii ARSEF4556 caused 100% mortality of pupae, larvae and adults on day 4, 6 and 12, respectively. This strain was further tested against adult weevils in different concentrations (10(5)-10(8) conidia cm(-2) or ml(-1)) using two types of fungal formulation: 'dry' conidia and 'wet' conidia (suspended in 0.03% aq. Tween 80) applied on different substrates (tissue paper, peat and Sitka spruce seedlings). 'Dry' conidia were more effective than 'wet' conidia on tissue paper and on spruce or 'dry' conidia premixed in peat. The LC(50) value for 'dry' conidia of isolate ARSEF4556 was three folds lower than 'wet' conidia on tissue paper. This study showed that 'dry' conidia are more effective than 'wet' conidia, causing 100% adult mortality within 12days. Possible strategies for fungal applications are discussed in light of the high susceptibility of larvae and pupae to fungal pathogen.     

Pathogenicity of Beauveria bassiana,Metarhizium anisopliae (Deuteromycotina: Hyphomycetes), and other entomopathogenic fungi against Lygus lineolaris (Hemiptera: Miridae)

The pathogenicity of 32 fungal isolates from the genera of Beauveria, Verticillium, Paecilomyces, Metarhizium, Mariannaea, and Hirsutella to second-instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), was tested under laboratory conditions. These isolates originated from various insect hosts and substrates from France, Denmark, Austria, Italy, Turkey, Syria, and the United States. A single exposure concentration (1 x 10(7) conidia/ ml) assay for each isolate was first conducted by immersing the insects in 10 ml of a fungal suspension for 5s. These were followed by concentration-mortality assays on five of the most pathogenic isolates using four test concentrations ranging from 2 x 10(4) to 2 x 10(7) conidia/ml. B. bassiana 726 (Bemisia-passaged GHA strain) was used as a standard for comparison in all of the assays. Among the test isolates, three produced mortality not significantly different from the water control. Mortality ranged from 35 to 98% among the other 29 isolates. The LC50 values of the five most pathogenic isolates ranged from 0.8 to 5.0 x 10(5) conidia/ml. The LT50 values for these isolates ranged from 6.0 to 6.9, 3.1 to 5.1, and 2.5 to 4.0 d for concentrations of 2 x 10(5), 2 x 10(6), and 2 x 10(7) conidia/ml, respectively. Two strains of B. bassiana (ARSEF 1394,5665) and one M anisopliae (ARSEF 3540) were more pathogenic to the nymphs than the standard, having significantly lower LC50 and LT50, values. Our results demonstrated that several genera of entomopathogenic fungi have promise as microbial control agents against L. lineolaris.     

Interactions of Metarhizium anisoplae and tree-based mulches in repellence and mycoses against Coptotermes formosanus (Isoptera: Rhinotermitidae)

The use of mulch in urban landscapes has increased in the United States for the past decade. Tree-based organic mulches can supply Coptotermes formosanus Shiraki with food, moisture, and shelter. The current research contributes to mulch management technology in termite control. A choice test arena was designed to determine the repellence and mortality caused by commercial mulches treated with different concentrations of the fungus Metarhizium anisopliae (Metschnikoff) against C. formosanus. Each of six tree-based mulches (pine bark, pine straw, bald cypress, eucalyptus, water oak, and melaleuca) was coated with six conidial concentrations ranging from 1 x 10(3) to 1 x 10(8) conidia/ml. The foragers of C. formosanus were repelled significantly by the fungal-treated mulch substrates; the proportion of termites on fungal-treated mulch was usually <20% during the 28-d test. By day 28, >99% of the termites were killed in test arenas containing a chamber with mulch treated with 10(7) or 10(8) conidia/ml. M. anisopliae significantly reduced mulch consumption by 34-71%. Mulch consumption by the termites was negatively correlated with fungal concentration, and the type of mulch also affected consumption. The differences in termite foraging activities, mortality, and food consumption among mulches were usually confounded by differences in fungal concentrations of M. anisopliae. The results indicate that repellence and virulence of M. anisopliae conidia should significantly reduce the suitability of these six mulches as a habitat for C. formosanus.     

Laboratory evaluation of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against puparia and adults of Ceratitis capitata (Diptera: Tephritidae)

Laboratory experiments were done to measure the pathogenicity of 10 autochthonous isolates of Beauveria bassiana (Balsamo) Vuill. and of five Metarhizium anisopliae (Metsch.) Sorok. toward puparia and adults of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Although all isolates applied via inoculation of the fungal suspensions on the ventral surface of the abdomen were pathogenic to adults, with mortality rates ranging from 30 to 100% and average survival times (ASTs) from 6.5 to 8.6 d, when C. capitata puparia were immersed in the conidial suspensions, only B. bassiana Bb-1333 and EABb 01/103-Su and M. anisopliae EAMa 01/58-Su isolates caused >50% mortality of puparia. In a second series of bioassays conducted on five selected isolates, adults were sprayed with four 10-fold concentrations ranging from 1.0 x 10(5) to 1.0 x 10(8) colony-forming units (cfu)/ml. The median lethal concentrations (LC50) of the four most virulent isolates ranged from 4.9 x 10(5) to 2.0 x 10(6) cfu/ml with estimated time to kill 50% of the insects ranging from 4.6 to 5.3 d. The effect of a sublethal dose (ca. LD50) of either B. bassiana EABb 01/103-Su or M. anisopliae EAMa 01/58-Su isolate was studied by reciprocal crossing. Treatment with B. bassiana reduced fecundity and fertility at 6, 8, and 10 d after treatment, with fecundity and fertility reductions ranging from 20.0 to 71.2% and from 33.6 to 60.0%, respectively. These reductions occurred in pairing combinations of treated females with either treated or nontreated males. M. anisopliae was more effective in reducing fecundity and fertility at 6 d after treatment, with the reduction varying from 58.4 to 72.1% and from 28.6 to 45.9%, respectively. In addition, the first oviposition was significantly delayed for 1 d in females treated by either fungal species. The above-mentioned five selected isolates were assayed against C. capitata puparia treated as late third instars in sterilized soil at 25'C under three moisture conditions (-0.1, -0.01, and -0.0055 MPa). At -0.01 MPa, all isolates were low pathogenic to C. capitata puparia, whereas significant differences in the puparia mortality occurred between isolates at -0.1 and -0.0055 MPa. The highest pupal mortalities ranged from 52.5 to 70.0%, as a function of soil moisture and were caused by EAMa 01/58-Su and Bb-1333 isolates.     

Synergistic effect of entomogenous fungi on some insecticides against Bihar hairy caterpillar Spilarctia obliqua (Lepidoptera: Arctiidae)

A number of fungal parasites infect a wide range of insects and cause epizootics from time to time. Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin are two of the major disease-causing fungi in insects. Investigations were carried out to study the effect of these fungi on the toxicity of endosulfan, imidacloprid, lufenuron, diflubenzuron, dimethoate and oxydemeton methyl against 10-11 days old larvae of Spilarctia obliqua (Walker). For some products the combination treatments showed higher dose mortality response than the sole treatment of fungal conidia or the insecticide. The combination of insecticides with B. bassiana showed 1.26-35.8 fold increase in toxicity of insecticides over sole treatment, while the increase was 1.05-72.0 fold in case of M. anisopliae. Imidacloprid 17.8 SL and oxydemeton methyl 25EC may be used in combination with these fungi for management of S. obliqua.     

A new bioassay method reveals pathogenicity of Metarhizium anisopliae and Beauveria bassiana against early stages of Capnodis tenebrionis (Coleoptera; Buprestidae)

We used a newly developed bioassay method to demonstrate for the first time the potential of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to be used for the control of neonate larvae of Capnodis tenebrionis, a major threat to stone-fruit orchards in several countries. Four B. bassiana and four M. anisopliae isolates were all pathogenic for neonate larvae of C. tenebrionis; mortality rates 10 days after inoculation by dipping in a suspension with 10(8)conidia/ml varied from 23.5% to 100%. Three of the four M. anisopliae isolates caused 100% mortality. In most cases, postmortem hyphal growth and sporulation of M. anisopliae or B. bassiana was observed covering the larvae in their galleries. The eight isolates were also evaluated for pathogenicity to C. tenebrionis eggs at the same dosage. Only two B. bassiana isolates caused significant egg hatching reduction of 84.5% and 94.5%. Our results indicate that M. anisopliae and B. bassiana may be considered as promising for a new approach to prevent larval infestations by C. tenebrionis.     

Pathogenicity of the Hyphomycete Fungi Verticillium lecanii and Metarhizium anisopliae to the Western Flower Thrips,Frankliniella occidentalis

The pathogenicity of several isolates of the hyphomycete fungi Verticillium lecanii and Metarhizium anisopliae to Frankliniella occidentalis was investigated. Treatment of adult thrips with M. anisopliae resulted in at least 94% mortality at 7 days post-inoculation. In contrast, V. lecanii isolates only gave mortalities of between 20 and 70%. Detailed studies were made on the most virulent isolate of M. anisopliae (275) to determine its efficacy at different doses and temperatures. At 23 C the LC was ca. 3 105 conidia ml-1 after 5 days and the LTs were 50 50 3 and 4.5 days at 10 7 and 106 conidia ml-1 respectively. Temperature influenced fungal virulence to adult thrips; the LT at 18 and 20 C was ca. 4 days and at 23 or 26 C it was ca. 3 days. 50 Larvae were less susceptible to infection than adult thrips (27% versus 100% mortality), presumably due to the inoculum being shed with the exuvium during ecdysis. Conidia of M. anisopliae isolate 275 germinated rapidly on the surfaces of larvae, pupae and adults, with most germlings producing appressoria within 24 h post-inoculation. Fungal elements were present in significant amounts in the body 3 days after treatment.     

Selection of entomopathogenic fungi for the control of the western corn rootworm Diabrotica virgifera virgifera

Abstract:  The western corn rootworm Diabrotica virgifera virgifera Le Conte (Col., Chrysomelidae), a serious pest of maize, has been recently introduced into Europe. Several approaches for its control are presently under investigation including microbial agents. During a field survey in Hungary in 2005, naturally occurring entomopathogenic fungi were found to attack this pest. These novel isolates together with standard isolates were tested for virulence against D. v. virgifera larvae and adults. Twenty strains of Metarhizium anisopliae , Beauveria bassiana and Beauveria brongniartii were used in bioassays in the laboratory. Larvae and adults were dipped into a spore suspension with a concentration of 1 × 10⁷ conidia (con.)/ml. They were kept for 14 days at 22°C (±2°C) and 70% relative humidity. The number of infected larvae and adults were counted and infection rates were calculated. Adults were significantly more susceptible to entomopathogenic fungi than larvae. The most virulent isolate infected about 47% of larvae ( M. anisopliae Ma2277), whereas the infection rate in adults was up to 97% ( M. anisopliae Ma2275). Isolates of M. anisopliae caused significantly higher mortalities than isolates of B. brongniartii and B. bassiana . Most of the adult beetles were killed within 12 days. Isolates from D. v. virgifera were more virulent than those from other hosts.     

Pathogenicity of entomopathogenic fungi to different developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae).

The pathogenicity of 4 species of entomopathogenic fungi (Hyphomycetes species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, and Paecilomyces fumosoroseus) to various developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae) ticks was compared under laboratory conditions. The most virulent isolate, M. anisopliae-108, caused 92-96% mortality to unfed larvae and nymphs on day 7 postinfection (PI) and 100% mortality to unfed adults and engorged females on day 21 PI. The pathogenicity of M. anisopliae-108 to engorged larvae and nymphs was lower--82.6 and 60%, respectively. All tested B. bassiana, M. flavoviride, and P. fumosoroseus isolates were significantly less virulent (P < 0.05) or avirulent toward most life stages of R. sanguineus. The M. anisopliae and M. flavoviride isolates also prevented or reduced the ability of the ticks to lay eggs several days before their deaths. Female ticks infected by the fungi achieved only 11.3-60.8% of their egg-laying capacity compared with the controls.     

Selection of a Highly Virulent Fungal Isolate,Metarhizium anisopliae Ma TB 101, for Control of Taro Beetle,Papuana uninodis (Coleoptera: Scarabaeidae)

Fungal isolates (31 Metarhizium anisopliae var. anisopliae , five M. anisopliae var majus , three Beauveria bassiana and four B. brongniartii ) originating from a wide range of geographical locations, insect species and soil types were tested against Papuana uninodis (Coleoptera: Scarabaeidae). In a first test series, spores were applied topically to third-instar larvae and adults. The most effective strain against P. uninodis larvae and adults was Ma TB 101, a M. anisopliae isolate from adult P. woodlarkiana . For adults, strain Ma FI 384, a M. anisopliae from Popillia japonica , was almost equally effective. The 11 most effective isolates (nine M. anisopliae var. anisopliae , one B. brongniartii and one B. bassiana ) with LT values of less than 6 weeks in 50 adults and/or less than 4 weeks in larvae were tested for their efficacy against adults and larvae of P. uninodis by application of spores to soil (107 spores/g). Ma TB 101 was significantly more effective against both adults and larvae (LT ca. 10 days) than all other isolates (LT > 50 50 3 weeks). Two other M. anisopliae strains, Ma F 248 from soil and Ma FI 384 from P.japonica , were more effective than most isolates in adults. The latter three M. anisopliae isolates were tested in a concentration series against third-instar larvae and adults. Mortality was concentration related. Isolates Ma F 248 and Ma FI 384 did not achieve 50% mortality within the test period at concentrations lower than 107 spores/g of soil or feed. For concentrations of Ma TB 101 from 1 107 to 2 105 spores/g the LT ranged from 13 to 30 days in adults and 12 to 24 50 days in third-instar larvae. Accordingly, concentrations causing 50% mortality (LC ) for Ma 50 TB 101 were significantly lower than for the two other M. anisopliae isolates tested.     

Susceptibility of Sitophilus zeamais (Motsch.) (Coleoptera: Curculionidae) and Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae) to Entomopathogenic Fungi from Ethiopia

The efficacy of 13 isolates of entomopathogenic fungi belonging to Beauveria , Metarhizium or Paecilomyces spp. was assessed against Sitophilus zeamais (Coleoptera: Curculionidae) and Prostephanus truncatus (Coleoptera: Bostrichidae) using a total immersion bioassay technique in the laboratory. Fungi were applied at concentrations of 1 ×10 7 and 1 ×10 8 conidia mL -1 for P. truncatus and S. zeamais , respectively. All isolates tested were virulent to P. truncatus (98-100% mortality, and median survival time (MST) ranged from 2.85-4.05 days). Metarhizium anisopliae and B. bassiana were also virulent to S. zeamais (92-100% mortality, MST ranged from 3.58-6.28 days). The isolate of Paecilomyces sp. was found to be the least virulent against S. zeamais , causing only 26.32 ±4.29% mortality with MST of 10.38 ±0.29 days. P. truncatus proved more susceptible to the entomopathogenic fungi tested than S. zeamais . One M. anisopliae (PPRC-EE) and three B. bassiana isolates (PPRC-HH, PPRC-9609 and PPRC-9614) were selected for further study and dose-mortality relationships were assessed on S. zeamais . The tested concentrations ranged from 1 ×10 4 -1 ×10 7 conidia mL -1 . M. anisopliae (PPRC-EE) showed the lowest LC 50 (3.39 ×10 5 conidia mL -1 ) followed by B. bassiana PPRC-HH (2.04 ×10 6 conidia mL -1 ). PPRC-9609 and PPRC-9614 showed slight differences in LC 50 but not at LC 90 . The results revealed the higher potency of M. anisopliae as compared with the B. bassiana isolates tested. The study suggests that the use of entomopathogenic fungi may hold promise as an alternative method to control pests of stored-products in Ethiopia.