Selection ofconidia/ml). Vegetative growth on culture media and sporulation on culture media, cooked rice and lesser mealworm cadavers were also evaluated. Isolates of B. bassiana were more effective than the M. anisopliae isolates and larvae were more susceptible than adults. The isolates UNIOESTE 04 and UNIOESTE 02 were originally obtained from adults of the lesser mealworm and were the most efficient isolates. In addition, UNIOESTE 04 showed high level of sporulation on different culture media and is considered a potential useful tool to lesser mealworm control.KEY WORDS: Biological control, poultry house, lesser mealworm RESUMO -Este trabalho teve por objetivo selecionar isolados de fungos entomopatogênicos, visando sua utilização no controle do cascudinho, Alphitobius diaperinus (Panzer). Larvas e adultos foram inoculados com suspensões de conídios de 99 isolados de Beauveria bassiana (Bals.) Vuill. e Metarhizium anisopliae (Metsch.) Sorok. em concentrações variáveis de 10 5 a 10 9

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.

-Trichogramma pretiosum Riley and the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae are effi cient biological control agents and are thought to be used jointly. In here, we investigated if these entomopathogens could have any side-effects on T. pretiosum. Therefore, 1 x 8 cards containing sterilized eggs of Anagasta kuehniella (Zeller) that were sprayed with 0.2 ml of B. bassiana or M. anisopliae (1.0 × 10 9 conidia/ml) were offered to a T. pretiosum female for 24h (30 cards/fungus = 30 replicates). Afterwards, females were isolated in glass tubes. The control group was sprayed with sterile distillated water + Tween 80 (0.01%). In addition, 60 cards with sterilized eggs of A. kuehniella were submitted to parasitism by females of T. pretiosum for 24h. Of these cards, 30 were sprayed with B. bassiana or M. anisopliae and 30 with distillated water + Tween 80 (0.01%), and observed daily until parasitoid emergence. Metarhizium anisopliae decreased parasitoid emergence and caused confi rmed mortality. Therefore, fi eld and semi-fi eld experiments should be conducted for a fi nal assessment of the side-effects of these entomopathogens on Trichogramma as a ways to develop a control strategy in which both can be used.

Knowledge about interactions among biological control agents, used in sequence or simultaneously, may help to increase their efficiency, or in the case of conservation biological control, to avoid the impairment of efficiency. The aim of this study was to evaluate the interaction between two isolates of Beauveria bassiana (Bals.) Vuill. (Hypocreales) (Unioeste 47 and Unioeste 57) and Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae), varying the timing of sprays of the fungus. Isolates were sprayed at 109 conidia ml−1 upon cards (1 × 5 cm; i.e., 2 × 105 conidia mm−2) with eggs of Anagasta kuehniella Zeller (Lepidoptera: Pyralidae) and the control group was sprayed with sterile distilled water with Tween® 80 (0.01%). First, the percentage of parasitized eggs was assessed in a choice test: females of T. pretiosum were confined with two cards, one of them treated with either of the fungus isolates, the other a water‐treated control. Second, parasitism was compared on cards sprayed at various time intervals pre‐ vs. post‐parasitism: the number of parasitized eggs was assessed, as well as the parasitoid's emergence percentage, egg‐adult period, offspring sex ratio, and the longevity of parasitizing females. Trichogramma pretiosum preferred to parasitize control eggs (65 and 72%) and parasitization of the treated eggs did not differ between fungus isolates: 35% (Unioeste 47) vs. 28% (Unioeste 57). Spraying before or after parasitizing did not affect the number of parasitized eggs, nor offspring sex ratio; however, both treatments affected the percentage of emerged adults, egg‐adult period, and longevity. These findings suggested that isolates did not interfere in the longevity of parasitizing females. When the fungus was sprayed on the various development stages of the parasitoid within the host egg, or when it was offered to the parasitoid at different times after spraying, differences in adults' longevity, parasitism, emergency period from egg to adult, and sex ratio were observed. Beauveria bassiana may be used in conjunction with this parasitoid in a program of pest control.

-Alphitobius diaperinus (Panzer), the lesser mealworm, is the most important pest in large-scale poultry production and is capable of harboring several types of poultry pathogens. Chickens feed readily on the beetles in infested litter and this sometimes causes nutritional problems and the weight gain may be affected. Chemical insecticides are usually applied, however beetle infestation are very difficult to control using these products. Besides, this control method can also offer risks to chicken and man. Microbial control may be a promising strategy to control A. diaperinus in poultry houses and natural occurrence of entomopathogenic fungi is very important to control the lesser mealworm in poultry houses. The objective of this work was to register the occurrence of Metarhizium anisopliae on adults of the lesser mealworm in commercial poultry houses in Brazil.KEY WORDS: Biological control, entomopathogenic fungi, animal production RESUMO -Alphitobius diaperinus (Panzer) é uma das pragas mais importantes que atacam criações comerciais de frango de corte em todo o mundo. Os insetos são hospedeiros de microrganismos patogênicos às aves, provocam ferimentos no trato digestivo das mesmas e afetam a conversão alimentar. Embora seja praticado, o controle dos insetos com inseticidas químicos é insatisfatório, na maioria das vezes, e oferece risco às aves. O controle biológico destaca-se como alternativa viável, uma vez que os entomopatógenos são inócuos a animais homeotérmicos, não oferecendo risco de contaminação às aves e ao produtor. A presença de entomopatógenos no ambiente contribui para o controle natural dos insetos-praga e, neste sentido, o presente trabalho teve como objetivo registrar a ocorrência natural de Metarhizium anisopliae sobre adultos de A. diaperinus, em aviários comerciais no município de Cascavel, PR. PALAVRAS-CHAVE: Controle biológico, fungo entomopatogênico, produção animal A avicultura de corte, no Brasil, representa um importante segmento agroindustrial, sendo uma das atividades mais dinâmicas e avançadas tecnologicamente, e está concentrada principalmente na Região Sul do Brasil, com destaque para as regiões oeste e sudoeste do estado do Paraná.Dentre os problemas associados a essa atividade destacase o cascudinho (Alphitobius diaperinus) (Panzer), visto que é responsável pela queda na conversão alimentar das aves, pois estas ingerem os insetos no lugar da ração balanceada.Os insetos são também transmissores de patógenos causadores de doenças nas aves (salmonela, vírus, fungos, protozoários e platelmintos parasitos), além de ferirem seu trato digestivo (Despins et al. 1994, Despins & Axtell 1995, McAlister et al. 1995.Os insetos permanecem em meio à cama do aviário (maravalha + excrementos) e, também no solo, principalmente ao redor dos comedouros onde há maior disponibilidade de alimento. Nos intervalos dados entre

The purpose was to evaluate the side effects of strains Metarhizium anisopliae (Metsch.) Sorokin sensu lato Unioeste 43 and M. anisopliae sensu stricto ESALQ 1641 on Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) under controlled conditions. A free-choice test for parasitism was performed, with the confinement of T. pretiosum females mated with cards (1 × 5 cm) containing age-standardized Anagasta kuehniella Zeller eggs, either sprayed with a fungal strain (109 conidia/ml) or 0.01% v/v Tween 80 (control). For the no-choice tests, T. pretiosum females mated were confined with cards sprayed with fungal strains before or after parasitism, and cards with fungal applications at different times. The number of parasitized eggs, percentage of emergence, longevity, egg-to-adult period, sex ratio, total and confirmed mortality by the fungus, and longevity of females that parasitized previously sprayed eggs, were assessed. Histological analysis of immature phases was also performed. The fungus was repellent to T. pretiosum in the free-choice test, while in the no-choice test, fungal applications before and after parasitism did not affect the number of eggs parasitized by T. pretiosum or the sex ratio of emerging adults. However, both strains affected adult emergence rates, the egg-to-adult period, and longevity. Overall, both M. anisopliae strains had minor effects on these biological parameters of T. pretiosum under controlled conditions. Hyphae were not detected in histological observations of immature stages of the parasitoid.

Bolsista de Produtividade em Pesquisa/CNPqNeotropical Entomology 35(1): 075-082 (2006) 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)ABSTRACT -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 (26ºC and 32ºC 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 26ºC (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 26ºC than adults stage. Regarding B. bassiana, differences on sensitivity between larval stages and adults were not observed at this temperature. However, at 32ºC, 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 26ºC) (P < 0,05).KEY WORDS: Entomopathogenic fungus, microbial control, animal production RESUMO -O estudo avaliou a influência da temperatura e de cama nova e usada na germinação, crescimento vegetativo, virulência e produção de conídios de isolados de Beauveria bassiana (Bals.) Vuill. e Metarhizium anisopliae (Metsch.) sobre larvas e adultos do cascudinho Alphitobius diaperinus (Panzer). O crescimento vegetativo e a produção de conídios foram avaliados em meio de cultura. Para a avaliação da virulência os insetos foram tratados com suspensões contendo 1 x 10 8 conídios/ ml. Os experimentos foram realizados em câmara B.O.D. (26ºC e 32ºC e 14h de fotofase). Diariamente os insetos mortos foram coletados para quantificação da produção de conídios. Para avaliação das camas, após a inoculação, insetos foram transferidos para recipientes contendo cama de aviário (nova e usada). Os isolados de B. bassiana foram mais sensíveis à temperatura elevada em relação aos isolados de M. anisopliae no que se refere à viabilidade, crescimento vegetativo e virulência (P < 0,05). A conidiogênese também foi maior para B. bassiana a 26ºC (7 a 11 x 10...

Natural Occurrence of Beauveria bassiana (Bals.) Vuilleman (Moniliales: Moniliaceae) on the Lesser Mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in a Poultry House in Cascavel, PR ABSTRACT -One of the main problems in poultry houses in Brazil is the lesser mealworm, Alphitobius diaperinus Panzer. This insect inhabits the litter, manure and soil of poultry houses, feeding on bird manure, cadavers and spilt feed. The lesser mealworm serves as a reservoir of many animal pathogens. Chickens also feed readily on the beetles in infested litter and this sometimes causes nutritional problems and affects weight gain. Current control measures are generally unsatisfactory and microbial control may be a promising strategy to control A. diaperinus. Natural occurrence of entomopathogens is very important for the natural control of insect pests and this paper records the occurrence of Beauveria bassiana (Bals.) Vuill. on the lesser mealworm in commercial poultry houses, in Brazil. The evaluation was carried out weekly for six months (September/2002 to February/2003. The soil around the poultry house was collected and transferred to Zoology Laboratory from Unioeste, at Cascavel, PR. Infected larvae and adults were always present, but infected pupae were registered twice. Larvae were more susceptible than pupae and adults (incidence ranging from 49% to 100%, mean 74%, 33% to 50%, mean 6.9%; 0.9% to 100%, mean 40.7%, respectively). The pathogenicity of the fungus was confirmed under laboratory conditions following procedures outlined in Koch's postulates. KEY WORDS: Microbial control, entomopathogenic fungus, animal production RESUMO -A presença de populações elevadas do cascudinho-dos-aviários, Alphitobius diaperinus Panzer, nos aviários é um dos principais problemas da avicultura brasileira. O inseto infesta os aviários, transmite doenças às aves e afeta seu desenvolvimento quando ingerido no lugar da ração. Seu controle é dificultado pelos hábitos crípticos e pela presença constante das aves nos aviários e o controle microbiano pode ser uma alternativa viável para o problema. Sabe-se que a presença de entomopatógenos no ambiente contribui para o controle natural dos insetos-praga. Este trabalho relata a ocorrência natural de Beauveria bassiana (Bals.) Vuill. sobre larvas e adultos do cascudinho em aviários comerciais no Brasil. O experimento foi realizado de setembro/2002 a fevereiro/2003, com amostragem semanal de insetos no solo periférico de um aviário comercial em Cascavel, PR. O fungo esteve presente em todas as avaliações, infectando principalmente larvas (49% a 100%, média de 74,4%) e adultos (0,9% a 100%, média de 40,7%) encontrados no solo, sendo que pupas infectadas ocorreram apenas em duas avaliações (33% a 50,0%, média de 6,9%). O isolado de fungo obtido nos cadáveres foi identificado como sendo B. bassiana e teve sua patogenicidade ao cascudinho comprovada pelo postulado de Koch, em condições de laboratório. PALAVRAS-CHAVE: Controle microbiano, fungo entomopatogênico, produção animal