Differentiation of Three Phenotypically Similar \u3ci\u3eBlattella\u3c/i\u3e spp.: Analysis with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism of Mitochondrial DNA

Differentiation of Blattella asahinai Mizukubo, Blattella vaga Hebard, and Blattella germanica (L.) was investigated using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). DNA fragments corresponding to mitochondrial CO1,CO1/TL2(small and large fragments), and ITS2 regions were amplified using PCR. The universal primers used for amplifying mitochondrial CO1 and CO1/TL2 (small and large fragments) were CO1-J-1718/CO1- N-2191, CO1-J-2441/TL2-N-3014, and CO1-J-1751/TL2-N-3014. Amplification was observed with all primer combinations, but diagnostic patterns were found only with AluI digested amplicons of CO1 and CO1/TL2 (large fragments) regions. Both inter/intraspecific polymorphism was observed for the 517 bp CO1 region between German (A, B) and Asian (A, B, C) cockroach populations. Polymorphic form B of the Asian cockroach (374 and 179 bp fragments) was nearly identical to the field cockroach (370 and 178 bp fragments). The CO1 amplicon (517 bp) differentiated German and field cockroaches but not Asian and field cockroaches. The CO1/TL2 amplicons (1275 bp) digested with AluI clearly differentiated all three species. The Asian cockroach had a distinct 540 bp fragment, whereas the field cockroach had a 403 bp fragment and the German cockroach lacked both fragments. Intraspecific polymorphs for CO1/TL2 amplicons (1275 bp) were observed with the German cockroach only. Portions of the CO1 and TL2 genes were sequenced, revealing 76% identity among German, Asian, and field cockroaches. Sequence data also revealed that German and Asian cockroaches were more closely related to each other than to the field cockroach

In Vivo Study on Combined Toxicity of \u3ci\u3eMetarhizium anisopliae\u3c/i\u3e (Deuteromycotina: Hyphomycetes) Strain ESC-1 with Sublethal Doses of Chlorpyrifos, Propetamphos, and Cyfluthrin Against German Cockroach (Dictyoptera: Blattellidae)

The effect of Metarhizium anisopliae (Metschnikoff) Sorokin strain ESC-1 alone and in combination with sublethal doses of commercial formulations of chlorpyrifos, propetamphos and cyfluthrin on mortality of CSMA strain of German cockroach, Blattella germanica (L.), was determined by conducting in vivo studies that included 3 bioassays. Spores of M. anisopliae cultured on SDAY media had germination of \u3e90%. Based on bioassay 1, doses ranging from 0.5 to 300 ppm of chlorpyrifos and propetamphos, and 0.05 to 40 ppm of cyfluthrin were selected for bioassays 2 and 3. Cockroach mortality ranged from 5 to 20% for insecticides alone and 48 to 70% for insecticides + M. anisopliae in bioassay 2. In bioassay 3, mortality ranged from 15 to 60% for insecticides and 57.5 to 92.5% for insecticides + M. anisopliae. Percentage of cockroach mortality resulting from insecticide +M. anisopliae combinations was significantly higher than insecticide alone. Mortality was also significantly higher in certain insecticide + M. anisopliae combinations than M. anisopliae alone. There was no significant interaction between M. anisopliae and insecticides with their concentrations in bioassay 2, indicating an additive effect. But in bioassay 3, a significant interaction was observed when M. anisopliae was combined with multiple insecticide concentrations. The interaction indicated an additive effect for chlorpyrifos and cyfluthrin, and a synergistic effect for propetamphos. There were significant differences in LT50 among various treatment combinations. M. anisopliae alone or insecticide + M. anisopliae combinations did not affect body weight in treated German cockroaches

Toxicity of Metarhizium anisopliae (Metchnikoff) Sorokin strain ESC-1 with and without selected insecticides to control German cockroach, Blattella germanica (L.)

Metarhizium anisopliae (Metchnikoff) Sorokin strain ESC-1 was virulent to German cockroach, Blattella germanica (L.). Topical application of M. anisopliae strain ESC-1 resulted in mortality ranging from 31.3 to 86.3% in 21 d. The lethal dose (LD 50) of M. anisopliae strain ESC-1 estimated for German cockroaches was 4.18 × 108 spores per milliliter. In vitro compatibility studies between insecticides plus M. anisopliae indicated compatibility with most of the insecticide concentrations used. There was no significant difference in the germination of conidia cultured on insecticide-amended SDAY media. But colony growth and sporulation was significantly affected by incorporation of insecticide to SDAY media. Colony growth was partially inhibited at 50 and 500 ppm of chlorpyrifos, and 500 ppm of propetamphos. There was no sporulation at 500 ppm of chlorpyrifos and propetamphos, and sporulation was significantly affected at 50 ppm of chlorpyrifos. Both additive and synergistic effects were observed in the in vivo compatibility study illustrating compatibility between the two agents. In general, joint application of insecticide plus M. anisopliae resulted in an additive effect. The only exception was 300 ppm of propetamphos where a synergistic effect was observed. Lethal time was significantly reduced in many of the insecticides plus M. anisopliae combinations by 50% or greater than half fold differences compared with M. anisopliae alone. Metarhizium anisopliae, insecticides, and insecticide plus M. anisopliae combinations did not affect the body weight of German cockroaches. There was no detectable level of destruxins identified from the culture filtrate of M. anisopliae strain ESC-1. Bioassay with destruxins by itself or in combination with sublethal doses of insecticides did not significantly increase mortality in German cockroaches

Virulence of \u3ci\u3eMetarhizium anisopliae\u3c/i\u3e (Deuteromycotina: Hyphomycetes) Strain ESC-1 to the German Cockroach (Dictyoptera: Blattellidae) and Its Compatibility with Insecticides

Virulence of Metarhizium anisopliae (Metschnikoff) Sorokin strain ESC-1 against the German cockroach, Blattella germanica (L.), was determined using 5 concentrations ranging from 83107 to 23109 spores per milliliter. The calculated LD50 value was 4.18 by 108 spores per milliliter (4.18 3 105 spores per cockroach). In vitro study was conducted to determine the compatibility of M. anisopliae strain ESC-1 with chlorpyrifos, propetamphos, and cyfluthrin. Insecticides did not affect conidial germination but did adversely affect the growth and sporulation of M. anisopliae strain ESC-1. The growth of M. anisopliae colonies on media amended only with 50 and 500 ppm of chlorpyrifos and 500 ppm of propetamphos treatments at 3, 6, and 9 d was significantly inhibited compared with the control. Similarly, sporulation was significantly reduced in treated colonies exhibiting partial colony growth. The colonies cultured on SDAY media amended with 50 ppm of chlorpyrifos had significantly reduced sporulation compared with the control and no sporulation was observed in colonies cultured on media amended with 500 ppm of chlorpyrifos and propetamphos