Michael G. Villani

Soil Moisture Effects on Entomopathogenic Nematodes

Abstract The effect of soil moisture on entomopathogenic nematode virulence was examined in the laboratory. Objectives were to determine the virulence of several species and isolates of entomopathogenic nematodes at various soil moisture contents and temperatures, and after fluctuations in soil moisture. Studies included up to five isolates of entomopathogenic nematodes: Heterorhabditis bacteriophora Poinar (Oswego and Tuscarora strains), Steinernema glaseri (Steiner) (NC1 strain), S. feltiae (Filipjev) (Biosys 369 strain), and S. carpocapsae (Weiser) (NY001 strain). Nematodes were applied to sandy loam soils ranging in soil moisture content from below the permanent wilting point of plants to near saturation. In all experiments, a rainfall or irrigation event was simulated by adding water to rehydrate soils to high moisture levels (near saturation). Nematode virulence was evaluated periodically by measuring insect mortality in Galleria mellonella (L.) larval bioassays, before and after rehydration. Nematode virulence increased with soil moisture content for all species and isolates tested. Our studies demonstrated that the virulence of entomopathogenic nematodes in low moisture conditions could be restored by rehydrating the soil. Insect mortality was generally low in low-moisture, nematode-infested soils before rehydration, but increased to high levels posthydration. Moisture effects were evident from the onset of each experiment, whereas the effect of soil temperature on nematode-induced insect mortality was delayed and nonsignificant until 14 wk after the initiation of the third experiment.

Identification and synthesis of female sex pheromone of Oriental beetle, Anomala orientalis (Coleoptera: Scarabaeidae)

Females of the Oriental beetle,Anomala orientalis (Waterhouse), release a sex pheromone composed of a 9:1 blend of (Z)- and (E)-7-tetradecen-2-one. The double-bond position of the pheromone was determined by DMDS derivatization and interpretation of the fragmentation patterns produced by monounsaturated ketones. In a sustained-flight tunnel, males responded by flying toward female beetles and attempting to copulate with them. Both effluvium and whole-body extracts of OB females were analyzed, and the activity was found only in the airborne extracts. Flight-tunnel bioassays also showed that a synthetic 90:10Z/E blend on a rubber septum was attractive and that the responses of males to this blend were equivalent toZ isomer alone, but much better than to the singleE isomer.

Essential amino acid methyl esters : Major sex pheromone components of the cranberry white grub, Phyllophaga anxia (Coleoptera : Scarabaeidae)

Chiral capillary gas chromatographic–electroantennographic detection (GC-EAD) analysis indicates that L-valine and L-isoleucine methyl esters are the major sex pheromone components released by females of the cranberry white grub, Phyllophaga anxia (LeConte). The GC retention times and GC-mass spectrometry of the two natural compounds were identical to those of authentic standards. Of five reproducible GC-EAD active components revealed with female volatiles, the L-valine and L-isoleucine methyl esters elicited the strongest male antennal responses. The ratio of L-valine and L-isoleucine methyl esters was determined to be 3:1 by analysis of pheromone gland extracts. Chirality was shown to be critical by GC-EAD, since only the L-form of these amino acid methyl esters elicited an EAD response. In field experiments conducted in Massachusetts, a synthetic 3:1 blend of L-valine and L-isoleucine methyl esters on a rubber septum was attractive to P. anxia males.

Trapping Phyllophaga spp. (Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada using sex attractants.

Abstract The sex pheromone of the scarab beetle, Phyllophaga anxia, is a blend of the methyl esters of two amino acids, L-valine and L-isoleucine. A field trapping study was conducted, deploying different blends of the two compounds at 59 locations in the United States and Canada. More than 57,000 males of 61 Phyllophaga species (Coleoptera: Scarabaeidae: Melolonthinae) were captured and identified. Three major findings included: (1) widespread use of the two compounds [of the 147Phyllophaga (sensu stricto) species found in the United States and Canada, males of nearly 40% were captured]; (2) in most species intraspecific male response to the pheromone blends was stable between years and over geography; and (3) an unusual pheromone polymorphism was described from P. anxia. Populations at some locations were captured with L-valine methyl ester alone, whereas populations at other locations were captured with L-isoleucine methyl ester alone. At additional locations, the L-valine methyl ester-responding populations and the L-isoleucine methyl ester-responding populations were both present, producing a bimodal capture curve. In southeastern Massachusetts and in Rhode Island, in the United States, P. anxia males were captured with blends of L-valine methyl ester and L-isoleucine methyl ester. Resumen La feromona sexual del escarabajo, Phyllophaga anxia, es una mezcla de los ésteres metílicos de dos aminoácidos, L-valina y L-isoleucina. Se condujo un estudio de campo usando diferentes mezclas de los dos componentes en 59 sitios de Estados Unidos y Canada. Más de 57,000 machos de 61 especies dePhyllophaga fueron capturados e identificados. Tres de los resultados más importantes incluyen: (1) el extenso uso de los dos componentes [de las 147 especies de Phyllophaga (sensu stricto), en Estados Unidos y Canada, fueron capturados machos de cerca del 40% de ellas.]; (2) para la mayoría de las especies, la respuesta intraespecífica de los machos a las combinaciones de los dos aminoácidos fue consistente entre años diferentes, y en todos los sitios geográficos; y (3) un inusual polymorfismo de la feromona fue descrito para P. anxia. Poblaciones de algunos sitios fueron atrapados sólo con valina, mientras que poblaciones de otros sitios fueron atrapados sólo con isoleucina. También se encontraron sitios donde las poblaciones responden a ambos componentes, valina e isoleucina, produciendo una curva de captura bimodal. En el sureste del estado de Massachusetts y en Rhode Island, en Estados Unidos, machos de P. anxia fueron atrapados en trampas con mezclas de valina e isoleucina.

Does host range influence susceptibility of herbivorous insects to non‐host plant proteinase inhibitors?

We examined the influence of proteinase inhibitors on digestive enzymes and development of oriental beetle, Exomala orientalis Waterhouse, European chafer, Rhizotrogus majalis (Razoumowsky), Phyllophaga white grub, Phyllophaga anxia (LeConte), cranberry root grub, Lichnanthe vulpina (Hentz), Japanese beetle, Popillia japonica Newman, Asiatic garden beetle, Maladera castanea (Arrow) (Coleoptera: Scarabaeidae), and the black cutworm, Agrotis ipsilon (Rottemburg) (Lepidoptera: Noctuidae). We demonstrated that all species within our test group had alkaline midguts that contained proteinase activity that could be inhibited, in vitro with serine proteinase inhibitors. Our data suggests that host range may influence the susceptibility to non‐host inhibitors. Chronic ingestion of the serine proteinase inhibitor, Kunitz‐soybean trypsin inhibitor (STI), significantly reduced proteolytic activity in vivo in those species with relatively specialized feeding habits (i.e., cranberry root grub, Japanese beetle, Asiatic garden beetle, and black cutworm). Chronic ingestion of STI also resulted in reduced larval growth and delayed pupation for black cutworm, and elevated larval mortality for Japanese beetle. However, chronic ingestion of STI did not influence larval survival for those species with relatively generalized feeding habits (i.e., oriental beetle, European chafer). Based on these results, we propose mechanistically‐based criteria for selecting proteinase inhibitors for phytochemical defense against herbivorous insects.

Spatial ecology of the Japanese beetle, Popillia japonica.

A cornerstone of spatial ecology is the quantification of the patchy nature of animal and plant populations in space. By using spatial covariance, total covariance, and quantile variance, we found that patchiness of Japanese beetle grub populations varied more between years than between fairways at a central New York golf course. We also observed that populations tended to shrink and swell around patches with perennially low density, that locations with perennially low grub density were associated with high soil organic matter content, and that locations with frequently high grub density were associated with intense adult beetle activity in the vicinity.

Methyl 2-(methylthio)benzoate: the unique sulfur-containing sex pheromone of Phyllophaga crinita

The female-produced sex pheromone of Phyllophaga crinita (Burmeister) (Coleoptera: Scarabaeidae: Melolonthinae; the adult has no common name) is identified as methyl 2-(methylthio)benzoate. This is the first identification of a sulfur-containing, long-distance, female-produced sex attractant from any insect taxa. The root-feeding larvae of this species are serious pests in many crops in Texas and Mexico. In field tests, many P. crinita males were captured in traps baited with the authentic compound. Interestingly, a heteroatom analog, methyl 2-methoxybenzoate, also captured P. crinita males, but only at a dose 10,000 times higher than the lowest tested dose of the authentic pheromone.

L-Leucine methyl ester: the female-produced sex pheromone of the scarab beetle, Phyllophaga lanceolata.

The female-produced sex pheromone of the scarab beetle Phyllophagalanceolata was identified as the methyl ester of an essential amino acid, l-leucine. During field testing, 239 male P. lanceolata were caught in traps baited with l-leucine methyl ester. l-Isoleucine and l-valine methyl esters, similar in structure to l-leucine methyl ester and previously identified as female-produced sex pheromone compounds employed by other Phyllophaga species, were also tested. Addition of l-valine or l-isoleucine methyl esters to the l-leucine methyl ester in 1:1 ratios completely inhibited attraction of P. lanceolata males. Males of P. squamipilosa were also captured using l-leucine methyl ester. This is the first record of P. squamipilosa from Kansas.

Identification of the Female-Produced Sex Pheromone of the Scarab Beetle, Hoplia equina

Hoplia equina LeConte (Coleoptera: Scarabaeidae: Melolonthinae) is a beetle pest of cranberry beds in Massachusetts. Larvae feed on the roots of the cranberry plant, reducing yield as well as vine density. The female sex pheromone was identified as 2-tetradecanone. There were eight compounds found in the airborne volatiles collected from females that elicited antennal responses from males. Of the eight compounds tested (nonanal, decanal, dodecanal, 2-dodecanone, 2-tridecanone, 2-tetradecanone, 2-pentadecanone, and 2-hexadecanone), 2-tetradecanone was the only one that attracted male beetles in the field. Combining any of the other seven antennally active compounds with 2-tetradecanone did not increase male capture.

Effect of soil environment on the efficacy of fungal pathogens against scarab grubs in laboratory bioassays

Abstract The effect of soil temperature and water on fungus-induced mortality of scarab grubs was investigated in the laboratory. Soil applications of dry mycelial particles of Metarhizium anisopliae (Metsch.) Sorokin were tested against the Japanese beetle, Popillia japonica Newman, and particles of Beauveria brongniartii (Saccardo) Petch were tested against the European chafer, Rhizotrogus majalis (Razoumowsky). Japanese beetle mortality occurred fastest in soil at 27°C and 11% water content and slowest in soil at 21°C and 17.5% water content. Japanese beetle mortality at Week 5 of the bioassay was affected by both soil temperature and water when a high dosage of fungus was used. The proportion of Japanese beetle cadavers with sporulating fungus was influenced by soil water, dose, and a soil temperature and dose interaction. In contrast, European chafer mortality was affected only by soil temperature. Mortality rates for B. brongniartii -killed European chafers and the proportion of cadavers supporting sporulating fungus were both significantly higher in soil at 21°C than at 27°C. Survival of M. anisopliae and B. brongniartii was affected by soil water; concentrations of both fungi were consistently higher in soil at low water content.