Field observations of the effects of fenitrothion and Metarhizium anisopliae var. acridum on non-target ground dwelling arthropods in the Sahel

The effect of the chemical insecticide, fenitrothion, and a mycoinsecticide based on Metarhizium anisopliae var. acridum on the activity of non-target epigeal arthropod scavengers was investigated in areas of open savannah in southeast Niger Republic, West Africa. Both insecticides were applied as full cover sprays to unreplicated 800 ha plots to assess their season-long control of Sahelian grasshoppers. Compared with control plots, fenitrothion caused an immediate but temporary reduction in grasshopper numbers, whereas M. anisopliae var. acridum provided delayed but prolonged control. Scavenging rates of pyrethroid-killed grasshoppers placed along transects in unsprayed plots and those treated with fenitrothion and M. anisopliae var. acridum at various intervals after spraying were assessed. In the fenitrothion plot, an immediate reduction in scavenging activity occurred that was still apparent after 40 days at the plot center, although recovery at the plot edges was more rapid. By contrast scavenging rates remained high over equivalent areas in the M. anisopliae var. acridum and two untreated plots. Concurrent to the scavenging study, counts of grasshopper cadavers resulting from the spray treatments were conducted. These counts revealed that the density of grasshopper cadavers remained low throughout the M. anisopliae var. acridum plot and explained <1% of the reduction in live grasshoppers resulting from treatment, compared with >20% in the fenitrothion plot. This shortfall in grasshopper cadavers resulting from the spray treatment in the M. anisopliae var. acridum plot was unexpected because in a monitoring study, fungus-killed (unlike pyrethroid-killed) grasshoppers were unattractive to scavengers and readily persisted in this plot, and thus should have become apparent. Given we did not observe significant grasshopper dispersal, the scarcity of cadavers generated in the M. anisopliae var. acridum plot, together with unquantified visual observations, suggests that predation of infected but living grasshoppers was high. Our data provide circumstantial evidence that the different effects of chemical and biological grasshopper control on grasshopper natural enemies may influence the efficacy of large-scale treatments.     

The mechanism of the mycoinsecticide diluent on the efficacy of the oil formulation of insecticidal fungus

Adverse conditions, including low humidity, UV irradiation, and high temperature, appreciably affect the efficacy of mycoinsecticides. Oil formulation increased the virulence of Metarhizium anisopliae var. acridum (Ascomycota: Hypocreales) against locusts and grasshoppers by reducing the dependence on saturated water. A mycoinsecticide diluent (a water-in-oil emulsion) has been widely used to dilute the oil formulation of M. anisopliae in China. The aim of our study was to elucidate the mechanism by which the mycoinsecticide diluent improves the virulence of M. anisopliae. We investigated the effects of the mycoinsecticide diluent on the virulence, invasion speed, and viability of the conidia under various adverse conditions. The results demonstrated that the mycoinsecticide diluent significantly improved the virulence of conidia at low humidity (68, 75, and 84%). In particular, at an RH of 68%, the LT₅₀ for locusts treated with the emulsion was 5.4 days and was 31.6% lower than the value for locusts treated with an oil formulation. In addition, the concentration of the hyphal bodies found in the haemolymph of locusts treated with emulsion was about 27-fold higher than that in locusts treated with oil formulation four days after inoculation. This result was further confirmed by determining the concentration of M. anisopliae var. acridum DNA in locust haemolymph using quantitative PCR. The percentage germination of conidia in the emulsion was also significantly higher than that in oil at 68% RH. There was no significant difference in percentage germination between conidia treated with the emulsion and oil when exposed to irradiation with ultraviolet-B (UV-B) or high temperature. These results demonstrate that the mycoinsecticide diluent enhances the virulence of M. anisopliae formulated in oil at low humidity by providing adequate water for germination without interfering with the UV tolerance and thermotolerance of the conidia.     

Behavioral thermoregulation in the migratory locust: a therapy to overcome fungal infection

We examined under laboratory conditions the thermopreference of the migratory locust, Locusta migratoria migratorioides, following infection by the entomopathogenic fungus Metarhizium anisopliae var. acridum and its influence on mycosis. Infected locusts raised their body temperature more frequently than healthy conspecifics through selection of high temperatures in a heat gradient. Thermoregulation did not, however, alter the frequency of feeding events nor the amount of food eaten by infected L. migratoria. A thermoregulation regime of a minimum of 4 h/day substantially increased survival of inoculated insects (by 85%). However, the therapeutic effect decreased when thermoregulation was delayed following inoculation of the pathogen. Thermoregulation reduced locust mortality but did not completely eliminate the fungus from infected hosts; the fungus grew and killed the insects when thermoregulation was interrupted. We suggest that periodic, short bouts of thermoregulation, when performed from the onset of infection and for an extended period of time, are sufficient to provide a therapeutic effect to infected hosts. Such thermoregulatory capacity of locusts may limit the potential of fungal pathogens as biological control agents under certain ecological conditions.     

Discovery of a North American genetic variant of the entomopathogenic fungus <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> var. <Emphasis Type="Italic">anisopliae</Emphasis> pathogenic to grasshoppers

A genetic variant of the entomopathogenic fungus Metarhizium anisopliae var. anisopliae, isolated from a soil in Alberta, Canada, from a location with a history of severe grasshopper infestations, was evaluated for pathogenicity in bioassays of living grasshoppers. Mortality in treated individuals drawn from a laboratory colony was 99% (LT₅₀ = 6.7 days, LT₉₀ = 9.6 days) at 12 days post-inoculation compared to 100% (LT₅₀ = 4.1 days, LT₉₀ = 5.8 days) mortality at 8 days in insects exposed to a commercial isolate of M. anisopliae var. acridum (IMI 330189). Experimental infection of field-collected grasshoppers under laboratory conditions with the native isolate of M. anisopliae var. anisopliae resulted in 100% (LT₅₀ = 4.4 days, LT₉₀ = 5.4 days) mortality attained within 7 days compared to 100% (LT₅₀ = 4.7 days, LT₉₀ = 6.3 days) mortality in 9 days in insects treated with M. anisopliae var. acridum. Amplification of fungal genomic DNA from the indigenous isolate with primers for the specific detection of M. anisopliae var. anisopliae produced a product almost 300 bp larger than expected based on previously known isolates. This is the first demonstration of a highly virulent, indigenous non-chemical control agent of grasshoppers in North America. GenBank Accession Nos. DQ342236, DQ342237.     

Thermal ecology of Zonocerus variegatus and its effects on biocontrol using pathogens

1 Thermal behaviour of the variegated grasshopper, Zonocerus variegatus, was investigated in the humid tropical zone of southern Benin, west Africa, in the dry seasons of 1996 and 1998. In 1998, investigations included studies of a population of grasshoppers sprayed with an oil‐based formulation of the entomopathogenic fungus Metarhizium anisopliae var acridum. 2 Body temperature measurements and observations of thermal behaviour both in the field and on thermal gradients in the laboratory, suggest that Z. variegatus was not an active behavioural thermoregulator. Although it did show shade‐seeking behaviour at high temperatures, no overt behavioural postures or microhabitat selection associated with heat gain and elevation of body temperatures was observed. Moreover, no alterations to thermal behaviour were found in response to infection by Metarhizium. 3 Body temperatures exhibited by Z. variegatus in the field will lengthen disease incubation of M. anisopliae var acridum compared with laboratory maintained, constant temperature conditions and may have a significant impact on pathogens with a lower thermal tolerance. 4 Habitat structure appeared to be an important factor determining the extent of body temperature elevation. The effect of habitat differences on infection and growth of M. anisopliae var acridum and other entomopathogenic fungi is discussed.     

Fever and phenotype: transgenerational effect of disease on desert locust phase state

Natural enemy attack can cause transgenerational shifts in phenotype such that offspring are less vulnerable to future attack. Desert locusts (Schistocerca gregaria) show density‐dependent variation in their resistance to pathogens, such that they are less vulnerable to pathogens when in the high‐density gregarious phase state (when they would probably be more exposed to pathogens) than when in the solitarious phase state. We therefore hypothesized that infected gregarious parents would maintain this phenotype in their offspring. We infected gregarious desert locust nymphs with the fungal pathogen Metarhizium anisopliae var. acridum, and allowed them to survive to reproduction by means of behavioural fever. The phase state of the locust offspring was assessed by their colouration and behavioural assays. Contrary to our hypothesis, we found an increase in solitarization in the infected population (14.6% solitarious offspring from infected parents, vs. <2% from uninfected counterparts at equivalent density). In a second experiment, we simulated behavioural fever temperatures and obtained a similar result (13.6% solitarious offspring vs. 4.4% from controls), implying that the phenomenon is probably a side‐effect of the hosts’ fever response. Identification of this novel environmental factor affecting locust phase state could have important implications for the biological control of these major pests.     

The inhibitory effect of the fungal toxin, destruxin A, on behavioural fever in the desert locust

During an infection locusts behaviourally fever by seeking out higher environmental temperatures. This behaviour places the pathogen at sub-optimal growth temperatures while improving the efficiency of the immune system, thereby prolonging the lifespan of the host. It is therefore in the interest of the pathogen to either adapt to fever-like temperatures or to evolve mechanisms to interfere with, or inhibit fever. We investigated the behavioural fever response of desert locusts to two fungal pathogens. A prolonged fever was observed in locusts infected with Metarhizium acridum. However, fever was comparatively short-lived during infection with Metarhizium robertsii. In both cases restriction of thermoregulation reduced lifespan. Destruxin A (dtx A) produced by M. robertsii, but not M. acridum has previously been associated with the inhibition of the insect immune system. Injection of dtx A during infection with the fever-causing M. acridum inhibited fever and was particularly effective when administered early on in infection. Furthermore, locusts injected with dtx A were more susceptible to M. acridum infection. Therefore engineering M. acridum isolates currently used for locust biocontrol, to express dtx A may improve efficiency of control by interfering with fever.     

Seasonal patterns of persistence and infectivity of Metarhizium anisopliae var. acridum in grasshopper cadavers in the Sahel

Field-based experiments were conducted to evaluate the fate and infectivity of the entomopathogenic fungus Metarhizium anisopliae var. acridum (Deuteromycotina: Hyphomycetes) in grasshopper cadavers in the Sahel. Unlike uninfected cadavers, which were rapidly scavenged, those infected with the fungus persisted in the environment for a number of weeks. The environmental factor most associated with cadaver disappearance was rainfall. The high environmental humidity associated with rainfall was also required for sporulation of the fungus on host cadavers, although the likelihood of sporulation differed between microsites. Characteristics of the infection profile from infective cadavers were investigated by the sequential exposure of uninfected hosts to sporulating cadavers in field cages. This experiment revealed that cadavers remained infective for > 30 days, with the net infectivity changing through time. The most likely explanation for these changes is climatic influences on both the fungus and host. High humidity was not required for infection. A measurement of the transmission coefficient between healthy hosts and sporulating cadavers in the field was obtained at a realistic density of infectious cadavers. This revealed a figure of 0.45 m² day^–1. Overall, these experiments show that following host death, M. anisopliae var. acridum can be persistent in the environment, sporulate on host cadavers and reinfect new hosts at a realistically low field density, although at least in arid or semi-arid areas, rainfall may be critical to the horizontal transmission of this pathogen.     

Field observations of the effect of Metarhizium anisopliae var. acridum on the saxaul locust,Dericorys albidula Serville (Orthoptera: Dericorythidae)

Dericorys albidula Serville (Orthoptera: Dericorythidae) is a major pest of Haloxylon ammodendron and other saxaul plant species in the Qom province, Iran. Using fungal insecticides can be an alternative method for chemical insecticides. Effect of insecticide fungi, Metarhizium anisopliae var. acridum, on the various nymphs of D. albidula was studied in the field through 2005 and 2006. Fixed concentrations of fungi (10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰ and 10¹³ spore mL^?1) were prepared as gasoline formulation and were sprayed on the locusts on H. ammodendron trees, and mortality percentage was recorded 15 days after treatment. The results showed that various concentrations significantly affected on the second, third, fourth and fifth nymphal instars of D. albidula compared to control in 2006, although this effect was lower in 2005 on nymphs. Mortality of the highest concentration (10¹³ spore mL^?1) was higher (17.6–24%) than other concentrations in all tests, but these values were not notable. The results of this study showed that using M. anisopliae var. acridum diluted in gasoline can be effective on nymphal instars of locust, D. albidula, in two continuous years.     

Development of a model for evaluating the effects of environmental temperature and thermal behaviour on biological control of locusts and grasshoppers using pathogens

1 Recent years have seen an upsurge in locust and grasshopper populations in many parts of the world. Environmentally sustainable approaches to locust and grasshopper control may be possible through the use of biopesticides based on entomopathogenic fungi. Unfortunately, the performance of these biopesticides is highly variable with environmental temperature and host thermoregulatory behaviour critically determining the pattern and extent of mortality after applications. Here, we present a temperature‐dependent model that enables us to predict the field performance of Metarhizium anisopliae var. acridum, the key fungal pathogen used in locust biopesticides. 2 The model was constructed using mortality rate data generated across a range of temperatures in the laboratory and is driven by environmental temperature data linked through host body temperature models. 3 Model predictions were validated against empirical field data obtained for five species, Locustana pardalina, Oedaleus senegalensis, Zonocerus variegatus, Nomadacris septemfasciata and Chortoicetes terminifera. Mortality predictions were accurate to a 2‐day error in every 10 days. This level of resolution is satisfactory to guide operational use of the biopesticide. 4 The model was subsequently used for a prospective evaluation of the performance of M. anisopliae var. acridum against two additional pest species, Dociostaurus maroccanus and Calliptamus italicus in Spain. Results suggest that this pathogen would work reasonably well against these species as long as early instars are targeted. 5 The model could provide a useful tool to assist in interpreting effectiveness of control operations, develop improved application strategies to optimize the performance of the biopesticide and identify appropriate target species and environments.