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.     

Effects of Metarhizium anisopliae on searching,feeding and predation by Orius albidipennis (Hem., Anthocoridae) on Thrips tabaci (Thy., Thripidae) larvae

The generalist predator Orius albidipennis Reut. (Hem., Anthocoridae) and entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Ascomycota: Clavicipitaceae) are important biocontrol agents of Thrips tabaci Lindeman (Thys., Thripidae) in most greenhouses. In this study, we estimated searching time, feeding time and predation rate in O. albidipennis feeding on untreated and treated thrips larvae which had been inoculated with three concentrations of M. anisopliae V275 at four time intervals; 0, 24, 48 and 72 h after infestation. Applied concentrations were 1×10³, 2×10⁴ and 2×10⁵ conidia/mL which roughly correspond to the LC₂₅, LC₅₀ and LC₇₅ for second instar-larvae. O. albidipennis responded to the presence of the M. anisopliae on the treated hosts by increasing the searching time and decreasing feeding time and predation rate levels. It was also able to detect and avoid treated patches.     

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.     

In vitro production of the biological control agent Heterorhabditis indica SL0708 in different agar media

ABSTRACT

Heterorhabditis indica SL0708 is an entomopathogenic nematode isolated from Valle del Cauca-Colombia, whose bacterial symbiont, Photorhabdus luminescens subsp. akhurstii SL0708, has potential to control pests of economic importance in Colombia. Since in vivo production does not supply its demand, this investigation evaluated H. indica SL0708 production on different agar media. Five culture media (I, II, III, IV and V) were evaluated for productivity and pathogenicity of infective juveniles (IJs). IJs emerged between 11 and 16 days after inoculation in all media, with a total of 2.7?×?10⁴ and 4.7?×?10⁶ IJs produced during 15 days after IJs emergence, with maximum productivity at day five and high variability. Pathogenicity to Galleria mellonella larvae was not significantly different between in vitro and in vivo produced IJs on all media tested. Media IV and V were selected for their higher productivity. Subsequently, nematode inoculum size was evaluated in selected media at 2000, 4000 and 6000?IJs ml^?1, but significant differences were not observed in productivity and pathogenicity. Lastly, lipid source influence was evaluated in medium IV comparing canola, olive and soy oils. None of the plant-based oils had a significant effect on IJs production and pathogenicity. A medium was selected for H. indica SL0708 IJs production which was suitable in terms of productivity, culture time and pathogenicity of IJs produced. The medium and parameters selected in this study could be applied as an alternative for mass production of this entomopathogenic nematodes.     

Potency of Bacillus thuringiensis and Bacillus subtilis against the cotton leafworm,Spodoptera littoralis (Bosid.) Larvae

The biological activities of two species of bacteria isolated from soil of cotton fields identified as Bacillus subtilis strain NRC313 (BS NRC313) and Bacillus thuringiensis strain NRC335 (BT NRC335) were evaluated against the third larval instar of the cotton leafworm, Spodoptera littoralis (Boisd.). The different entomopathogenic bacteria of BS NRC313and BT NRC335 contained 10 × 10⁸ cell/ml, and caused mortality of 100 and 97.3% for the above mentioned strains, respectively. Concentrations of 2.5 × 10⁸ to 10 × 10⁸ cell/ml of strains BS NRC313 and BT NRC335 were applied to the larvae: LC₅₀ were 3.3 × 10⁸ and 3.9 × 10⁸ cell/ml respectively. The influence of exposure to toxin concentrations manifested in terms of decreasing the adult emergence and prolongation of the generation period. The percentage of larvae that survived and succeeded to pupate increased by decreasing the concentration. The longevity of adult emergence that resulted from larvae treated with Bacillus subtilis were 6.0 ± 0.51 and 9.0 ± 0.63 days at 5 × 10⁸ and 2.5 × 10⁸ cell/ml, respectively compared with 9.8 ± 0.47 in control. The results indicated that Bacillus subtilis was more potent than Bacillus thuringiensis. Field applications of B. thuringiensis, B. subtilis and Reldan achieved 55.6, 67.4 and 89.4% reduction of the cotton leafworm larvae Spodoptera littoralis in clover plants under field conditions.     

Influence of cell density and phase variants of bacterial symbionts (Xenorhabdus spp.) on dauer juvenile recovery and development of biocontrol nematodes Steinernema carpocapsae and S. feltiae (Nematoda: Rhabditida)

The rhabditid nematodes Steinernema carpocapsae and Steinernema feltiae are used in biological control of insect pests. Mass production is done in liquid culture media pre-incubated with their bacterial symbionts Xenorhabdus nematophila and Xenorhabdus bovienii, respectively, before nematode dauer juveniles (DJs) are inoculated. As a response to food signals produced by the bacterial symbionts, the DJs exit from the developmentally arrested dauer stage (they recover development) and grow to adults, which produce DJ offspring. Variable DJ recovery after inoculation often causes process failure due to non-synchronous population development and low numbers of adult nematodes. This contribution investigated the influence of the bacterial cell density on DJ recovery and development to adults. At higher density of 10¹⁰ bacterial cells ml⁻¹, a higher percentage of DJ recovery was induced, and adults occurred earlier in both Steinernema spp. than at lower density of 10⁹ and 10⁸ cells ml⁻¹. Xenorhabdus symbionts produce phase variants. Recovery in bacteria-free supernatants was lower than in supernatants containing bacterial cells for both primary and secondary phase Xenorhabdus spp. and lower in secondary than in primary phase supernatants or cell suspensions. In general, recovery was lower for Steinernema feltiae and the time at which 50% of the population had recovered after exposure to the food signal was longer (RT₅₀ = 17.1 h) than for Steinernema carpocapsae (RT₅₀ = 6.6 h). Whereas >90% S. carpocapsae DJs recovered in hemolymph serum of the lepidopteran insect Galleria mellonella, recovery of S. feltiae only reached 31%. Penetration into a host insect prior to exposure to the insect’s food signal did not enhance DJ recovery. Consequences for liquid culture mass production of the nematodes and differences between species of the genera Steinernema and Heterorhabditis are discussed.     

Selection of Antonospora locustae (Protozoa: Microsporidae) with higher virulence against Locusta migratoria manilensis (Orthoptera:Acrididae)

Antonospora locustae is a microsporidian parasite of grasshopper insects that is used as a biological control agent. We report on laboratory selection of isolates from different regions with increased virulence. Bioassays were conducted against third instar nymphs of Locusta migratoria manilensis. AL2008L01 was originally imported from the USA in 1986, AL2008M01 was isolated from Melanoplus differentialis in USA and AL2008F01 was isolated from infected Fruhstorferiola tonkinensi collected in Guangdong, China. The results showed that all three isolates can infect the locust and that pathogenicity increased gradually with increased dose. The LD₅₀ values of the original isolates at the highest dose (5×10⁶ spores/nymph) were 19, 23 and 22 days and LD₅₀ values were 3.2×10⁵, 3.4×10⁶ and 0.7×10⁶ spores/g, respectively. After selecting for three generations, the virulence of all isolates increased significantly. LT_50s were reduced to 17, 20 and 21 days at the highest dose (5×10⁶ spores/nymph) and LD_50s were reduced to 1.4×10⁵, 2.5×10⁵ and 1.7×10⁵ spores/g.     

<Emphasis Type="Italic">Beauveria bassiana</Emphasis> pathogenicity to the cherry slugworm, <Emphasis Type="Italic">Caliroa cerasi</Emphasis> (Hymenoptera: Tenthredinidae) larvae

The cherry slugworm Caliroa cerasi is a significant destructive pest of sour cherries (Prunus cerasus) in Turkey. The potential of entomopathogenic fungi for controlling C. cerasi was evaluated. The effects of exposure methods and conidial concentrations (1 × 10⁶, 1.5 × 10⁶, 1 × 10⁷ and 1.5 × 10⁷ conidia/ml) on mature larvae of C. cerasi infected by Beauveria bassiana were investigated under laboratory conditions. Larvae sprayed directly with B. bassiana conidial suspensions and larvae exposed to B. bassiana-treated leaves resulted in 100% mortality within 2.90 and 2.77 days, respectively. The median lethal time (LT₅₀) and days to mortality were highest in the 1.0 × 10⁷ concentrations of conidia for both direct spray and leaf exposure. The present study suggests that B. bassiana has good potential for control of the cherry slugworm, C. cerasi.     

Quantitative and qualitative study of the bacterial flora of farmed freshwater prawn (Macrobrachium rosenbergii) larvae

Quantitative and qualitative studies of the bacterial flora of farmed freshwater prawn (Macrobrachium rosenbergii) larvae in Saudi Arabia were performed, and isolates identified where possible. Physico‐chemical characteristics, bacterial counts, and the nature of the bacterial flora of larvae rearing tank water, sediment, tank wall surfaces, larval surface, supplied water, and feed were investigated. Bacterial counts ranged from 2.1 ± 1.3 × 10⁵ to 2.2 ± 0.8 × 10⁷ colony forming units (CFU) ml^?1 in tank water; 4.4 ± 0.9 × 10⁷ to 8.3 ± 1.7 ×10⁹ CFU g^?1 in tank sediment; 8.6 ± 1.0 × 10² to 9.8 ±0.7 × 10⁴ CFU cm^?2 on the tank wall surface; 1.3 ± 1.1 × 10⁴ to 7.7 ± 1.6 × 10⁶ CFU per larva surface, 7.9 ± 1.2 × 10⁵ to 5.0 ± 1.5 × 10⁷ CFU g^?1 in washed larval tissue slurries, 9.1 ± 0.7 × 10³ CFU ml^?1 in supplied water, and 2.4 ± 1.9 ×10¹⁰ CFU g^?1 in mixed feed. Fourteen bacterial genera were identified, including Chryseomonas sp., Vibrio spp., Cellulomonas sp., Aeromonas hydrophila, and Pasteurella sp. The tank water and sediment had similar bacteria to those on the prawn larvae. Chryseomonas sp., Cellulomonas sp. and Vibrio sp. were the most dominant species (prevalence >10%) in tank water; Chryseomonas sp., Pseudomonas alcaligenes and Shewanella putrefaciens in the sediment; Ps. alcaligenes and Cellulomonas sp. on the tank wall surface; Chryseomonas sp., and Cellulomonas sp. on the larval surface; and Chryseomonas sp., Vibrio vulnificus, Sh. putrefaciens and V. alginolyticus in the washed larval tissue slurries (prevalence 10%). Pseudomonas alcaligenes, Moraxella sp., Serratia liquefaciens, Gordona sp. and Burkholderia glumae were absent in larvae but identified in the culture water, tank sediment, and tank wall surface. Pseudomonas sp., Chryseomonas sp., Pasteurella sp. and V. alginolyticus were the prevalent bacteria (>12%) in supplied water. The feed contained V. alginolyticus, A. hydrophila and Cellulomonas sp. as the dominant bacteria (>13%). In the culture water and larvae samples, 83% of the feed and supplied water bacteria were identified.     

Laboratory evaluation of Turkish entomopathogenic nematodes for suppression of the chestnut pests,Curculio elephas (Coleoptera: Curculionidae) and Cydia splendana (Lepidoptera: Tortricidae)

The lepidopteran, Cydia splendana, and the coleopteran, Curculio elephas, are the most serious pests of chestnut fruit in Turkey. We evaluated the biological control potential of three Turkish entomopathogenic nematode species, Steinernema feltiae, S. weiseri and Heterorhabditis bacteriophora, against the last instar larvae of C. splendana and C. elephas, both of which occur in the soil from fall (October–November) until mid-summer (August). The optimal temperature for infection, time to death of the hosts, and reproductive potential of the nematodes were determined at 10, 15, 20 and 25°C for both pest species. Cydia splendana was more susceptible to nematode infection than C. elephas. Temperature had a significant effect on the infectivity and development of entomopathogenic nematodes. The cold-adapted S. weiseri and S. feltiae were the most virulent species at 10 and 15°C, whereas the warm-adapted H. bacteriophora was the most effective at 20 and 25°C. In soil pot experiments conducted at 15°C, S. weiseri was the most virulent species against C. elephas and C. splendana. However, our data show that C. elephas larvae had a lower and C. splendana larvae had a higher susceptibility to the nematode species tested. Accordingly, we recommend that future efforts of using entomopathogenic nematodes, especially S. weiseri, be directed against C. splendana and that there be a continued effort to find more virulent nematode isolates against larvae of C. elephas.     

Infectivity of Steinernema mushtaqi (Rhabditida: Steinernematidae) against insect pests and their mass production

The infectivity of entomopathogenic nematode (EPN), Steinernema mushtaqi was tested against legume pod borer, Maruca vitrata, tobacco caterpillar, Spodoptera litura, blue butterfly, Lampides boeticus, red hairy caterpillar, Amsacta moorei, brown bug, Clavigralla gibbosa, mealy bug, Centrococcus somatics, fruit borer, Earias vittella and green bug, Nezara viridula larvae and in vivo mass production of the above-tested species of EPN have been carried out during 2008. S. mushtaqi was found to be more pathogenic to A. moorei, as it brought about 100% mortality within 48 h, than to S. litura, L. boeticus, N. viriduala and E. vittella, as mortality occurred within 72 h; whereas this level of mortality was recorded in C. somatis, C. gibbosa and M. vitrata within 144 h. No mortality was observed in the control treatment. Multiplication of S. mushtaqi and the yield of infective juveniles (IJs) on these insects was the highest (0.94 × 10⁵ IJs/cadaver) from N. viriduala, followed by S. litura (0.76 × 10⁵ IJs/cadaver), L. boeticus as also C. gibbosa (0.31 × 10⁵ IJs/cadaver) and M. vitrata (0.20 × 10⁵ IJs/cadaver). Very poor populations of IJs were found from A. moorei (0.15 × 10⁵ IJs/cadaver) and C. somatics (0.01 × 10⁵ IJs/cadaver). No multiplication of IJs was found from the cadaver of E. vittella. This opens a new hope of utilising S. mushtaqi in the insect pests management programme.     

Susceptibility of Phthorimaea operculella Zeller (Lepidoptera; Gelechiidae) to its granulovirus PoGV with larval age

1 Granulovirus PoGV is a strong candidate to substitute for chemical insecticides in integrated pest management (IPM) of the potato tuber moth Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae). Generally, susceptibility to baculoviruses decreases with increasing larval age. For effective field applications, knowledge of the age‐related change in susceptibility is crucial. 2 The relative susceptibility of various instars of P. operculella larvae to PoGV was studied by leaf‐disc bioassays in the laboratory at 26 °C incubation temperature. The LC₅₀ increased from 4 × 10⁶ granules/mL for neonate larvae to 1.2 × 10⁷, 1.1 × 10⁸, 4.4 × 10⁸, and 4.2 × 10⁹ granules/mL, signifying resistance ratios of 3, 29, 110, and 1051, for 5, 6, 7, and 9‐day‐old larvae, respectively. 3 The relationship between log‐LC₅₀ values and log‐larval weight was significantly linear. A logistic function described significantly the relation between larval weights and physiological age measured in degree‐days (DD > 13.4 °C). Both functions allowed prediction of the activity ratio of PoGV for different larval ages by using temperature summation to express physiological age. 4 PoGV was found to be highly active against P. operculella larvae up to a physiological age of approximately 50 DD (>13.4 °C) (i.e. the first third of the total larval development time). Thereafter, the virus rapidly lost its activity against older larvae. Prospects for applying this knowledge in the field are discussed.     

Insecticidal efficacy of two isolates of Beauveria bassiana (Bals.) (Vuill.), on the second larval stage of Leptinotarsa decemlineata (Say) (Col.: Chrysomelidae)

Leptinotarsa decemlineata is a serious potato pest throughout the world that has developed resistance to many insecticides. This study investigated the virulence of two indigenous isolates of Beauveria bassiana, consisting of DEBI007 and IR1217C. Five different concentrations of each isolate, 1 × 10⁷, 3.1 × 10⁷, 1 × 10⁸, 3.1 × 10⁸ and 1 × 10⁹ conidia ml^?1, were applied in bioassay using dipping method on second instar larvae. Control insects were treated with distilled water containing 0.01% Tween-80 (Sigma, Germany) as surfactant. The results showed that mortality percentages due to DEBI007 and IR1217C isolates in the highest concentration were 60% and 57.77%, respectively. There was a significant difference between efficacies of different conidial concentrations; therefore, with rise in conidial concentration, the mortality percentage was increased. The DEBI007 isolate that was previously isolated from the soil was more effective than IR1217C isolate. Calculated LC₅₀ for this effective isolate was 1.4 × 10⁷ conidia ml^?1, which, respectively, is the highest concentration.     

Characterization of the <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Isolates Virulent Against Rice Leaf Folder, <Emphasis Type="Italic">Cnaphalocrocis medinalis</Emphasis> (Guenee)

Soil samples were collected from different rice fields of Singur, Hooghly, West Bengal, India. Spore forming bacteria were isolated from the soil samples and among them, two isolates (BUSNC25 and BUSNC26) were larvicidal against third, fourth and fifth instar larvae of rice leaf folder, Cnaphalocrocis medinalis. The phenotypic, biochemical characterization and 16S rDNA analysis of the two isolates were done. On the basis of phenotypic, biochemical and phylogenetic analysis, the selected bacterial isolates (BUSNC25 and BUSNC26) were identified as Bacillus thuringiensis. The antibiotic sensitivity tests of these two isolates against selected doses of some standard antibiotics were done. Against the 3rd, 4th and 5th instar larvae of C. medinalis, the LC₅₀ values of BUSNC25 were 2.45 × 10⁴, 1.325 × 10⁴ and 2.35 × 10⁴ cfu/ml and of BUSNC26 were 3.375 × 10⁴, 1.9 × 10⁴ and 3.325 × 10⁴ cfu/ml, respectively.     

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.     

Influence of inoculum density on population dynamics and dauer juvenile yields in liquid culture of biocontrol nematodes Steinernema carpocapsae and S. feltiae (Nematoda: Rhabditida)

For improvement of mass production of the rhabditid biocontrol nematodes Steinernema carpocapsae and Steinernema feltiae in monoxenic liquid culture with their bacterial symbionts Xenorhabdus nematophila and Xenorhabdus bovienii, respectively, the effect of the initial nematode inoculum density on population development and final concentration of dauer juveniles (DJs) was investigated. Symbiotic bacterial cultures are pre-incubated for 1 day prior to inoculation of DJs. DJs are developmentally arrested and recover development as a reaction to food signals provided by their symbionts. After development to adults, the nematodes produce DJ offspring. Inoculum density ranged from 1 to 10 × 10³ DJ per milliliter for S. carpocapsae and 1 to 8 × 10³ DJs per milliliter for S. feltiae. No significant influence of the inoculum density on the final DJ yields in both nematode species was recorded, except for S. carpocapsae cultures with a parental female density <2 × 10³ DJs per milliliter, in which the yields increased with increasing inoculation density. A strong negative response of the parental female fecundity to increasing DJ inoculum densities was recorded for both species with a maximum offspring number per female of >300 for S. carpocapsae and almost 200 for S. feltiae. The compensative adaptation of fecundity to nematode population density is responsible for the lack of an inoculum (or parental female) density effect on DJ yields. At optimal inoculation density of S. carpocapsae, offspring were produced by the parental female population, whereas S. feltiae always developed a F1 female population, which contributed to the DJ yields and was the reason for a more scattered distribution of the yields. The F1 female generation was accompanied by a second peak in X. bovienii density. The optimal DJ inoculum density for S. carpocapsae is 3–6 × 10³ DJs per milliliter in order to obtain >10³ parental females per milliliter. Density-dependent effects were neither observed on the DJ recovery nor on the sex ratio in the parental adult generation. As recovery varied between different batches, assessment of the recovery of inoculum DJ batches is recommended. S. feltiae was less variable in DJ recovery usually reaching >90%. The recommended DJ inoculum density is >5 × 10³ DJs per milliliter to reach >2 × 10³ parental females per milliliter. The mean yield recorded for S. carpocapsae was 135 × 10³ and 105 × 10³ per mililiter for S. feltiae.     

Isolation and characterization of a Serratia marcescens with insecticidal activity from Polyphylla olivieri (Col.: Scarabaeidae)

Members of the genus Serratia are known for their abilities to infect insects. In this study, a red‐pigmented S. marcescens was isolated and characterized from the infected larvae of Polyphylla olivieri using bacterial cultivation, phylogenetic analysis as well as bioassays against larvae of the two insect pests, Plodia interpunctella and Ephestia kuehniella. Comparative 16S rRNA and groEL gene sequence BLAST analyses strongly suggested that the isolated strain should be placed in the genus Serratia, sharing high sequence similarities with several strain of S. marcescens associated with insects. Phylogenetic analysis placed the isolated bacterium with other S. marcescens bacteria in a clade with high bootstrapping values. To assess pathogenicity of the S. marcescens isolate, the bacterial cells were either injected into the haemolymph of the fifth‐instar larvae or added to the diets of insects. Survival curves of the control insects and those challenged with six different concentrations of S. marcescens showed that the S. marcescens isolate significantly reduced survival rates of the larvae. The LC₅₀s of the bacterium on P. interpunctella and E. kuehniella were 1992.26 and 1.09 × 10⁴ (CFU/μl) for injection bioassays at 6 h post‐injection, and 4.48 × 10⁴ and 1.96 × 10⁵ (CFU/10 μl) for feeding bioassays at 24 h post‐feeding, respectively. Injection of the bacterial culture supernatant into the larvae led to continuous bleeding from the site of injection, while injection of heat‐treated culture supernatant of the bacterium did not cause continuous bleeding. Together, our results showed the possibility of using this S. marcescens isolate in microbial control of the insect pests after addressing the safety concerns. Moreover, it might be considered as a source of useful bioactive molecules and genes with application in insect control and biotechnology via developing insect‐resistant plants.     

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.     

First record of Aspergillus oryzae (Eurotiales: Trichocomaceae) as an entomopathogenic fungus of the locust,Locusta migratoria (Orthoptera: Acrididae)

A novel entomopathogenic fungus of Locusta migratoria was identified as Aspergillus oryzae using a comparative sequence analysis of the internal transcribed spacer regions, aflatoxin B₁ detection and morphological analysis. The fungus isolated from a dead locust collected in northwestern China was found to be pathogenic to the insect. Phylogenetic experiments revealed a 99% similarity between the fungus and those of three species, A. oryzae, Aspergillus flavus and Aspergillus parvisclerotigenus which are in the same branch of the Flavi section of the genus Aspergillus. Tests to detect aflatoxin B₁ demonstrated that this fungus is a non-aflatoxin B₁ producer, unlike A. parvisclerotigenus. Furthermore, morphological comparison with A. oryzae and A. flavus revealed that Aspergillus sp. XJ-1 belongs to A. oryzae, and named as A. oryzae XJ-1. The results of bioassays against third-instar locusts showed that mortality was dose-dependent and its median lethal concentrations were 3.3 × 10⁸, 1.7 × 10⁷ and 7.2 × 10⁶ conidia/ml on the 10th-, 13th- and 15th-day post-inoculation. Therefore, the A. oryzae XJ-1 may have biocontrol potential against locusts.     

Entomopathogenicity of beauveria bassiana and metarhizium anisopliae to the cowpea aphid,aphis craccivora koch (homoptera: Aphididae)

The pathogenicity of four isolates each of the entomopathogenic fungi, Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) Sorok. to apterous adult Aphis craccivora Koch was evaluated in the laboratory at 4 concentrations of conidia. All fungi isolates tested were found to be pathogenic to the insect but their virulence varied among species and isolates within species. Three isolates, B. bassiana CPD 11 and M. anisopliae CPD 4 and 5 caused significantly higher mortality than the other isolates at the various concentrations tested causing mortality of between 58–91%, 64 to 93% and 66–100%, respectively, at 7 days post treatment. At the highest concentration of 1 × 10⁸conidiaml^‐1, these isolates produced the shortest LT₅₀s of 3.5, 3.6 and 3.4 days, respectively. Their LC₅₀s were 6.8 × 10⁵, 3.1 × 10⁵ and 2.7 × 10⁵ conidia ml^‐1, respectively. The results indicate that these isolates are promising candidates for the control of the cowpea aphid but their pathogenicity to various aphid non‐target beneficial organisms within the cowpea agroecosystem warrant further investigation before initiating field control.