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The aim of this study was to investigate the effect of starch granules enriched with carvacrol and mixed with straw pellets (as poultry litter) on the mortality of larvae and adults of the lesser mealworm, Alphitobius diaperinus Panzer, a cosmopolitan pest inhabiting chicken houses in vast numbers worldwide. Additionally, the effect of starch granules on the growth parameters and survival of broiler chickens exposed to treated litter was examined. In this study, granules containing 3, 5, and 10% carvacrol was used. In a simulated chicken house bioassay, this material was mixed with pellets in three different proportions: 30/70%, 40/60%, and 50/50% (granules/pellets, respectively). On this medium, young larvae (approximately 10 days old), older larvae (last stage before pupa), and unsexed 7–10 days old adults of the lesser mealworm, with access to food, were colonized. Experiments were performed at 29°C in the dark. The study shows that poultry litter with the addition of starch granules enriched with 10% of carvacrol in the proportion of 40:60% (granules:pellets) appears to be the optimal medium applicable to broiler houses for A. diaperinus control. In this environment, all larvae and adults died within 3–4 days and the overall development of the experimental chickens was similar to that of the control. However, the feed conversion rate was slightly higher in the treated group (1.72) than in the control group (1.56). The average final body weight in the treated group was 100 g lower than that in the control group (the differences were not statistically significant).

Upon extracting soluble proteins from insects as potential food ingredient, endogenous enzymes, such as phenoloxidases, are expected to negatively affect protein properties. The effect of phenoloxidases on solubility and digestibility of proteins was investigated for larvae of Tenebrio molitor, Alphitobius diaperinus and Hermetia illucens. Phenoloxidase inhibition was done using blanching (50 s, 90 °C) before extraction or extracting in presence of sulfite. Similar soluble protein yields and compositions were found without and with sulfite addition, whereas blanching decreased soluble protein yield. Upon in-vitro hydrolysis by pepsin and trypsin, soluble proteins from H. illucens were more digestible than those of T. molitor and A. diaperinus. Phenoloxidase activity during grinding negatively affected in-vitro pepsin hydrolysis. Besides phenoloxidase activity, also endogenous proteases were shown to remain active at pH 8 in extracts containing sulfite and after blanching of larvae. This stresses that protease activity needs to be carefully controlled in the design of insect based ingredients.

This study focused on the coagulation behavior of protein from larvae of Alphitobius diaperinus. The effect of incremental CaCl2 concentration (10, 15, 20 and 20 mmol/L) and temperature (90, 100 °C) on physical-chemical properties of insect coagula was investigated. A yield between 76 and 83 g of coagulum was obtained from 100 g of fresh larvae, decreasing with higher temperature and CaCl2. Protein-protein interactions and microstructure of coagula were analyzed respectively by means of protein solubility, SDS-PAGE and SEM. When higher temperature was applied, hydrophobic interactions and disulphide bonds increased due to a larger degree of protein denaturation, thereby contributing to the formation of large protein aggregates. Thus, significant increase in hardness of the coagula was observed, with specimens at 20 mmol/L CaCl2 being more than twice harder at 100 °C than at 90 °C. Moreover, proteins homologous to actin and tropomyosin contributed to the coagulum structure by hydrophobic interactions, whereas hemolymph proteins formed disulphide bonds. Increasing concentration of CaCl2 from 10 to 20 mmol/L, at 100 °C, displayed a smoother network that increased coagula hardness from 1200 to 2900 g respectively. Results of this study provide important information for the product development in relation to insect protein-based meat analogues.

This study analysed the effectiveness of using different levels of a product based on 6% cypermethrin and 2% citronella on broiler production and control the lesser mealworm (Alphitobius diaperinus). A total of 648 one-day-old chicks (Cobb®) which were allocated to one of three treatments (no application of the commercial product - control, and two concentrations of the product 2.00 and 3.33g/m2) in eight replicates with 27 birds each. The commercial product, which was based on cypermethrin was applied in the chambers with an atomizer. The lesser mealworm population was sampled weekly (1, 7, 14, 21, 28, 35 and 42 days of the experiment) using traps, at three points (front, middle and back) inside the pen. The performance, carcass and cut yields were evaluated. At 21, 28, 35 and 42 days of the experiment, the number of lesser mealworm adults and larvae was lower in the treatment groups involving 2.00 and 3.33g/m2 of the product than in the control group. On the first and 14th days of the study, the number of lesser mealworms adults was lower in the group treated with 3.33g/m2 of the product compared to the control group, although the results did not differ statistically from those obtained with 2.00g/m2 of the product. In conclusion, the application of the commercial product at both concentrations (2.00 and 3.33g/m2) was effective in combating Alphitobius diaperinus adults and larvae, and the performance was not influenced by treatments. RESUMO: Este estudo analisou a eficácia do uso de diferentes níveis de um produto à base de cipermetrina a 6% e citronela a 2%, na produção de frangos de corte e no controle do cascudinho (Alphitobius diaperinus). Um total de 648 pintos de um dia de idade (Cobb®) foram distribuídos em três câmaras com os tratamentos (sem aplicação do produto comercial - controle, e duas concentrações do produto comercial - 2,00 e 3,33g/m2) em oito repetições com 27 aves cada. O produto comercial a base de cipermetrina foi aplicado nas câmaras com um atomizador. A população de cascudinho foi amostrada semanalmente (1, 7, 14, 21, 28, 35 e 42 dias do experimento) usando armadilhas, em três pontos (frente, meio e fundo) de cada parcela. Foram avaliados o desempenho, rendimentos de carcaça e cortes. Aos 21, 28, 35 e 42 dias do experimento, o número de adultos e larvas de cascudinho foi menor nos grupos de tratamento envolvendo 2,00 e 3,33g/m2 do produto quando comparado ao grupo controle. No primeiro e no 14º dia do estudo, o número de cascudinhos adultos foi menor no grupo tratado com 3,33g/m2 do produto em comparação ao grupo controle, embora os resultados não tenham diferido estatisticamente daqueles obtidos com 2,00g/m2 do produto. A conversão alimentar de 1 a 21 dias de idade foi significativa, com os melhores resultados para essa variável quando aplicado 3,33g do produto por metro quadrado e no tratamento controle. Em conclusão, a aplicação do produto comercial em ambas as concentrações (2,00 e 3,33g/m2) foi eficaz no combate a adultos e larvas de Alphitobius diaperinus e, não influenciou o desempenho.

Lesser mealworm (Alphitobius diaperinus) is an insect species that can be reared on an industrial scale for human consumption. In this study, the hydration behavior of processed lesser mealworm food ingredients (whole powder, protein concentrate, textured protein) was analyzed in comparison to whey protein concentrate and tofu. Moisture adsorption isotherms were determined gravimetrically over 0.11–0.96 water activity range at 5 and 20 °C, and seven sorption models were applied to fit the experimental data. The Guggenheim-Anderson-de Boer model performed best on the isotherms of all studied samples. The determined adsorption isotherms were type III according to Brunauer classification. Mealworm protein concentrate presented the strongest hydration capacity at 20 °C, followed by whey protein concentrate and whole mealworm powder. Textured mealworm protein and tofu exhibited similar hydration behavior at 5 °C. Moreover, browning and agglomeration of mealworm protein concentrate powder were observed at water activities of 0.73 and higher, which can be explained by its composition and porous structure and were not typical for whey protein concentrate. The moisture adsorption results are important for the prediction of lesser mealworm ingredients shelf-life or prevention of undesirable quality changes during storage, but more research on microbial growth and sensory quality is needed.

The lesser mealworm, Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae), is the most important pest in poultry production systems worldwide. Adults and larvae serve as alternative low-quality food for chickens and as a vehicle for animal pathogens resulting in significant economic losses for farmers. Control of this pest is usually performed through insecticide application during the interval between chicken flocks. However, due to the cryptic behaviour of this pest, chemical control is usually not effective. With the aim of developing a sustainable approach to control A. diaperinus, we evaluated integration of the A. diaperinus aggregation pheromone with a highly virulent strain (Unioeste 04) of the fungus Beauveria bassiana (Bals.) Vuill. (Ascomycota: Cordycipitaceae) using an attract-and-infect device. Significant levels of A. diaperinus mortality and lower numbers of offspring (larvae) were observed in laboratory experiments using Unioeste 04. Horizontal transmission of the Unioeste 04 strain through contaminated living A. diaperinus and cadavers was also observed. In field experiments in commercial poultry houses, a significant level of A. diaperinus mortality was observed in Unioeste 04-treated arenas. The results show that the A. diaperinus aggregation pheromone can be used as an attractant in a device impregnated with infective fungal propagules, increasing adult infection and enhancing disease spread. In addition, the use of the microbial control agent inside the device allows the fungus to remain effective for longer periods in the adverse conditions of the poultry house environment.

Reducing Alphitobius diaperinus in poultry production is a difficult task. However, attempts should be made to control the insect pest, as it poses a serious threat to the life and health of the chickens, as well as the workers on a farm. Our research was conducted in two stages to assess the effectiveness of the biocidal paint against A. diaperinus, containing active substances such as permethrin and a mixture of ultramarine and violet 23. In the first stage, under laboratory conditions, after 22 days, 100% mortality of A. diaperinus larvae and adults was achieved. This allowed us to assume that the biocidal paint may also be effective in poultry houses. In the poultry house where biocidal paint was applied, the number of insects decreased continuously alongside the sampling dates. In both research stages, the biocidal paint proved more effective against A. diaperinus than traditional limewash, and also the time to effective interaction of the paint was noted. Additionally, it was observed that the larvae were more susceptible to the active substances than adults. The research was practical, however, further analyses are necessary to fully control A. diaperinus, especially in poultry houses.

Insects are gaining interest as an alternative protein source for feed/food purposes. Although the lesser mealworm (LM) is commercially produced for human consumption, published data on its nutrient composition is scarce. This study reports on LM larvae reared on 18 different diets composed of side-streams to (1) determine the nutritional composition of the larvae and (2) study the effect of dietary changes on the larval nutrient composition. The LM larvae proved to be of good nutritional value with essential amino acids profiles comparable with that of beef and linoleic acid (C18:2) was the most dominant essential fatty acids in the larvae. The side-stream based diets varied on dry matter basis in protein (16&ndash, 34%) and lipid content (2&ndash, 19%). The nutrient content of the larvae reared on diets that supported good growth ranged between 37% and 49% of protein, 22% and 26% of lipid and 4% to 6% of chitin on dry matter basis. No significant correlations were identified between the larval protein or lipid content and that of the diet, but it was found between the diet nutrients and larval growth. Based on larval growth data and economic considerations, diets composed of wheat middlings with a 10&ndash, 15% inclusion of rapeseed meal were identified as suitable feed for LM. Highest larval yields were obtained with diets containing 15&ndash, 22% of proteins and 5&ndash, 10% of lipids.

The use of plastics burgeoned in the last decades to become an essential component of our society. An environment friendly method to dispose of plastic waste is not available yet, to the outcome that these accumulate in landfills or are scattered as microplastics. New researches reported that some coleopteran species are able to destroy plastics thanks to their chewing mouthparts and the metabolic activity of their gut microbiota. This study shows that the lesser mealworm Alphitobius diaperinus is capable of feeding on, and apparently degrading, polystyrene. The gut microbiota of polystyrene-fed larvae was characterized using an NGS metagenomic approach, targeting both bacteria and fungi. Several microbe taxa emerged as differentially abundant between treatment and control groups (Cronobacter, Kocuria and Pseudomonas as bacteria, Aspergillus, Hyphodermella, Trichoderma as fungi). Some of them have been found in association with plastic compounds and/or have been proposed to be capable of plastic degradation. This research supports the notion that, although synthetic molecules, unlike most natural compounds, do not generally enter the natural food chain to be degraded by the environmental microbiota, some microbial communities may be able to decompose plastics. We speculate that, once identified, such communities may open to the possibility of devising bioreactors for plastic degradation.

The lesser mealworm, Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae), is considered the primary insect pest in broiler farms in Brazil. In this study, we characterized the susceptibility of A. diaperinus populations from broiler farms of southern Brazil to cypermethrin and chlorpyrifos. Larvae and adults of A. diaperinus were exposed to these technical insecticides diluted in acetone in residual bioassays. A geographic variation in the susceptibility of larvae and adults of A. diaperinus to both insecticides was detected. The larval LC50 for cypermethrin ranged from 0.43 to 7.33 µg a.i./cm2. Two populations from Santa Catarina state presented higher resistance ratios of 13.6- and 17-fold. When adults were exposed to cypermethrin, the LC50 ranged from 0.46 to 4.93 µg a.i./cm2, with population SC-3 from Santa Catarina having lower susceptibility (resistance ratio of 10.7-fold). When exposed to chlorpyrifos, A. diaperinus larvae present LC50 values ranging from 0.21 to 4.30 µg a.i./cm2. Larvae from Paraná and Santa Catarina (SC-1 population) presented the highest resistance ratios, ranging from 10- to 20-fold. In adults, the LC50 of chlorpyrifos ranged from 0.17 to 5.30 µg a.i./cm2, showing a maximum resistance ratio of 31-fold in a population from Paraná state. Based on LC99 values, candidate diagnostic concentrations of 15 and 12 µg a.i./cm2 of cypermethrin and chlorpyrifos, respectively, were also estimated for the resistance monitoring of A. diaperinus in Brazil. The implications of these results in Insect Resistance Management are discussed.

Insects are considered a nutritionally valuable source of alternative proteins, and their efficient protein extraction is a prerequisite for large-scale use. The protein content is usually calculated from total nitrogen using the nitrogen-to-protein conversion factor (Kp) of 6.25. This factor overestimates the protein content, due to the presence of nonprotein nitrogen in insects. In this paper, a specific Kp of 4.76 ± 0.09 was calculated for larvae from Tenebrio molitor, Alphitobius diaperinus, and Hermetia illucens, using amino acid analysis. After protein extraction and purification, a Kp factor of 5.60 ± 0.39 was found for the larvae of three insect species studied. We propose to adopt these Kp values for determining protein content of insects to avoid overestimation of the protein content.

This paper describes a methodology to evaluate Alphitobius diaperinus (Darkling beetle) population in order to estimate the effectiveness of control methods, consisting of direct sample collections from the poultry litter in 18 points, counting of adults and larvae, and subsequent comparison of the values obtained before and after of control strategies. It allows estimating the total population, population in a known area, as well to analyze the variations in different points all over the aviary. The method has shown to be very versatile, being indicated for population studies and efficacy evaluations, once it has already been validated in different experimental conditions.

Background: Increased amino acid availability stimulates muscle protein synthesis (MPS), which is critical for maintaining or increasing muscle mass when combined with training. Previous research suggests that whey protein is superior to soy protein in regard to stimulating MPS and muscle mass. Nevertheless, with respect to a future lack of dietary protein and an increasing need for using eco-friendly protein sources it is of great interest to investigate the quality of alternative protein sources, like insect protein. Objective: Our aim was to compare the postprandial amino acid (AA) availability and AA profile in the blood after ingestion of protein isolate from the lesser mealworm, whey isolate, and soy isolate. Design: Six healthy young men participated in a randomized cross-over study and received three different protein supplementations (25 g of crude protein from whey, soy, insect or placebo (water)) on four separate days. Blood samples were collected at pre, 0 min, 20 min, 40 min, 60 min, 90 min, and 120 min. Physical activity and dietary intake were standardized before each trial, and participants were instructed to be fasting from the night before. AA concentrations in blood samples were determined using 1H NMR spectroscopy. Results: A significant rise in blood concentration of essential amino acids (EAA), branched-chain amino acids (BCAA) and leucine was detected over the 120 min period for all protein supplements. Nevertheless, the change in AA profile was significantly greater after ingestion of whey than soy and insect protein (p &lt, 0.05). Area under the curve (AUC) analysis and AA profile revealed comparable AA concentrations for soy and insect protein, whereas whey promoted a ~97% and ~140% greater AUC value than soy and insect protein, respectively. A tendency towards higher AA concentrations beyond the 120 min period was observed for insect protein. Conclusion: We report that ingestion of whey, soy, and insect protein isolate increases blood concentrations of EAA, BCAA, and leucine over a 120 min period (whey &gt, insect = soy). Insect protein induced blood AA concentrations similar to soy protein. However, a tendency towards higher blood AA concentrations at the end of the 120 min period post ingestion was observed for insect protein, which indicates that it can be considered a &ldquo, slow&rdquo, digestible protein source.

Insects are increasingly used as a dietary source for food and feed and it is therefore important to understand how rearing conditions affect growth and development of these agricultural animals. Temperature is arguably the most important factor affecting metabolism and growth rate in insects. Here, we investigated how rearing temperature affected growth rate, growth efficiency and macronutrient composition in two species of edible beetle larvae: Alphitobius diaperinus and Tenebrio molitor.Growth rates of both species were quantified at temperatures ranging from 15.2 to 38.0 degrees C after which we measured protein and lipid content of the different treatment groups. Metabolic rate was measured in a similar temperature range by measuring the rate of O-2 consumption (VO2) and CO2 production (VCO2) using repeated measures closed respirometry. Using these measurements, we calculated the growth efficiency of mealworms by relating the energy assimilation rate to the metabolic rate. Maximum daily growth rates were 18.3% and 16.6% at 31 degrees C, for A. diaperinus and T. molitor respectively, and we found that A. diaperinus was better at maintaining growth at high temperatures while T. molitor had superior growth at lower temperatures. Both species had highest efficiencies of energy assimilation in the temperature range of 23.3-31.0 degrees C, with values close to 2 J assimilated/J metabolised in A. diaperinus and around 4J assimilated/J metabolised in T. molitor. Compared to "conventional" terrestrial livestock, both species of insects were characterised by high growth rates and very high energy conversion efficiency at most experimental temperatures. For A. diaperinus, lipid content was approximately 30% of dry mass and protein content approximately 50% of dry mass across most temperatures. Temperature had a greater influence on the body composition of T. molitor. At 31.0 degrees C the lipid and protein content was measured to 47.4% and 37.9%, respectively but lipid contents decreased, and protein contents increased when temperatures were higher or lower than 31.0 degrees C.In summary, rearing temperature had large and independent effects on growth rate, energy assimilation efficiency and protein/lipid content. Accordingly, temperature is a critical parameter to control in commercial insect rearing regardless if the producer wants to optimise production speed, production efficiency or product quality.

Diatomaceous earth (DE) is an inert powder obtained from milling the fossilized deposits of diatoms. Mainly composed of silica dioxide, DE is highly hygroscopic and is recommended for use in poultry houses as an evironmental conditioner, with a secondary action being the reduction of lesser mealworm populations Alphitobius diaperinus (Panzer). This study aimed to characterize and assess the insecticidal activity of a new environmental conditioner for poultry houses (Poultry Sec®, Vet Science Bio Solution, Maringá, Paraná, Brazil) in the laboratory. The study was conducted by adding DE to poultry feed (1, 2 and 3 g DE/kg feed), as well as by applying the DE to the poultry litter (86 and 172 g DE/m2), with A. diaperinus adults (26°C in the dark for 10 days). DE samples were analyzed by optical microscopy to measure and count the frustules and to identify the genera of the components. The insecticidal action of the product was observed. When applied to the litter, the DE caused between 83 and 85% mortality. With respect to the qualitative analysis of the diatoms, we estimated the concentration of 1.2 × 106 individuals/g of the product, distributed in nine distinct genera, with Brachysira Kützing being the predominant genus (40%).

The main aim of the presented paper is to assess the potential repellent effect of selected essential oils (EOs) against the lesser mealworm (Alphitobius diaperinus), which can cause economic losses in storage and in the poultry industry. Due to the development of pesticide resistance in A. diaperinus populations, as well as an attempt to limit extensive use of potentially harmful pesticides in food-related industries, there is a strong need for the development of alternative methods of dealing with A. diaperinus infestations. Because of their cost-effectiveness, availability and low vertebrate toxicity, EOs are promising agents in pest management. In the presented paper four off-the-shelf EOs: mint, vanilla, lemon and citronella (and mixtures of them) were tested as potential repellents. Moreover, a novel preference assay, providing an extended analysis of the preference and the locomotor response, was used. The most effective EOs were: citronella and lemon. EOs mixtures were generally more repellent than individual EOs, with the lemon and vanilla 1:1 mixture acting as the strongest repellent. A few of the tested EOs caused significant alterations to the locomotor activity, although no direct relation was observed. In conclusion, EOs can be potentially used as repellent agents in A. diaperinus management. Additionally, data on the locomotor activity may lead to designing better push-pull strategies in pest management.

Susceptibility of Alphitobius diaperinus (Panzer) (Coleoptera, Tenebrionidae) to cypermethrin, dichlorvos and triflumuron in southern Brazil. The lesser mealworm, Alphitobius diaperinus (Panzer), is an important insect pest in poultry houses in Brazil. Susceptibility of the lesser mealworm collected from eight poultry houses in Paraná state, southern Brazil, was evaluated for cypermethrin, dichlorvos and triflumuron. Adult A. diaperinus were tested in bioassays with cypermethrin and dichlorvos. Larvae were fed rabbit feed wetted with a triflumuron-water solution. Concentration-mortality regressions were estimated using Probit analysis and resistance ratios were calculated based on the susceptible population. Among the field populations evaluated, cypermethrin LC50 values for adults, ranged from 68.1 to 6,263 ng (AI)/cm². LC50 values for adults challenged with dichlorvos ranged from 10.3 to 1,385 ng (AI)/cm². One population from Pato Branco showed reduced susceptibility to triflumuron (LC50 = 272 µg (AI)/ml of solution) when compared to the most susceptible population (LC50 = 109.8 µg (AI)/ml). Application of cypermethrin and dichlorvos analogues should be managed with caution to minimize insecticide resistance problems. Suscetibilidade de Alphitobius diaperinus (Panzer) (Coleoptera, Tenebrionidae) a cipermetrina, diclorvós e triflumurom no sul do Brasil. O cascudinho, Alphitobius diaperinus (Panzer), é uma importante praga em aviários no Brasil. A suscetibilidade do cascudinho à cipermetrina, diclorvós e triflumurom foi avaliada em oito aviários do Estado do Paraná, Brasil. Adultos de A. diaperinus foram testados mediante bioensaios com cipermetrina e diclorvós. As larvas foram alimentadas com ração para coelhos impregnada com suspensões de triflumurom. As regressões de concentração-mortalidade foram estimadas usando análise de Probit e as razões de resistência calculadas com base na população suscetível. Entre as populações de campo avaliadas, os valores da CL50 para adultos tratados com cipermetrina variaram entre 68,1 to 6.263 ng (IA)/cm². Os valores da CL50 para adultos tratados com diclorvós variaram de 10,3 to 1.385 ng (IA)/cm². Uma população de Pato Branco apresentou reduzida suscetibilidade ao triflumurom (CL50 = 272 µg (IA)/ml), quando comparada à população mais suscetível (CL50 = 109,8 µg (IA)/ml de solução). Cipermetrina, diclorvós e análogos devem ser manejados e aplicados com cautela para minimizar os problemas de resistência a inseticidas.

This study aimed to investigate the potential accumulation of mycotoxins in the lesser mealworm (Alphitobius diaperinus, LMW) and black soldier fly (Hermetia illucens, BSF) larvae. Feed was spiked with aflatoxin B1, deoxynivalenol (DON), ochratoxin A or zearalenone, and as a mixture of mycotoxins, to concentrations of 1, 10, and 25 times the maximum limits set by the European Commission for complete feed. This maximum limit is 0.02 mg/kg for aflatoxin B1, 5 mg/kg for DON, 0.5 mg/kg for zearalenone and 0.1 mg/kg for ochratoxin A. The mycotoxins and some of their metabolites were analysed in the larvae and residual material using a validated and accredited LC-MS/MS-based method. Metabolites considered were aflatoxicol, aflatoxin P1, aflatoxin Q1, and aflatoxin M1, 3-acetyl-DON, 15-acetyl-DON and DON-3-glycoside, and α- and β-zearalenol. No differences were observed between larvae reared on mycotoxins individually or as a mixture with regards to both larvae development and mycotoxin accumulation/excretion. None of the mycotoxins accumulated in the larvae and were only detected in BSF larvae several orders of magnitude lower than the concentration in feed. Mass balance calculations showed that BSF and LMW larvae metabolized the four mycotoxins to different extents. Metabolites accounted for minimal amounts of the mass balance, except for zearalenone metabolites in the BSF treatments, which accounted for an average maximum of 86% of the overall mass balance. Both insect species showed to excrete or metabolize the four mycotoxins present in their feed. Hence, safe limits for these mycotoxins in substrates to be used for these two insect species possibly could be higher than for production animals. However, additional analytical and toxicological research to fully understand the safe limits of mycotoxins in insect feed, and thus the safety of the insects, is required.

Two cDNAs encoding different acetylcholinesterase (AChE) genes (AdAce1 and AdAce2) were sequenced and analyzed from the lesser mealworm, Alphitobius diaperinus. Both AdAce1 and AdAce2 were highly similar (95 and 93% amino acid identity, respectively) with the Ace genes of Tribolium castaneum. Both AdAce1 and AdAce2 have the conserved residues characteristic of AChE (catalytic triad, intra‐disulfide bonds, and so on). Partial cDNA sequences of the Alphitobius Ace genes were compared between two tetrachlorvinphos resistant (Kennebec and Waycross) and one susceptible strain of beetles. Several single nucleotide polymorphisms (SNPs) were detected, but only one non‐synonymous mutation was found (A271S in AdAce2). No SNPs were exclusively found in the resistant strains, the A271S mutation does not correspond to any mutations previously reported to alter sensitivity of AChE to organophosphates or carbamates, and the A271S was found only as a heterozygote in one individual from one of the resistant A. diaperinus strains. This suggests that tetrachlorvinphos resistance in the Kennebec and Waycross strains of A. diaperinus is not due to mutations in either AChE gene. The sequences of AdAce1 and AdAce2 provide new information about the evolution of these important genes in insects. Arch Insect Biochem Physiol. © 2007 Wiley‐Liss, Inc.

O objetivo deste trabalho foi avaliar a interferência dos métodos de bioensaios nos resultados de seleção de fungos entomopatogênicos, com a utilização de quatro isolados de Beauveria bassiana e adultos de Alphitobius diaperinus. Quanto à forma de inoculação, foram testados os seguintes métodos: imersão do inseto em suspensão de conídios; pulverização da suspensão sobre insetos; e tratamento de superfície com a suspensão em oito concentrações. Para avaliar a interferência do tempo de imersão sobre a mortalidade, os insetos foram colocados em suspensão de conídios por 1, 10, 20, 30, 40, 50 e 60 segundos. Conídios produzidos em arroz, insetos e meio sintético foram utilizados para avaliar a influência do substrato na virulência dos isolados. Os isolados UNIOESTE 4 e CG 152 foram os mais virulentos pelos métodos imersão e pulverização, respectivamente; em tratamento de superfície, a mortalidade foi baixa para todos os isolados. Observou-se que a mortalidade dos insetos é proporcional ao tempo em que ficam imersos, com tendência a se estabilizar após 40 segundos. Os isolados CG 71 e CG 152, produzidos sobre insetos, foram menos virulentos. Os métodos utilizados podem influenciar os resultados de bioensaios de seleção de fungos para o controle de insetos. The objective of this work was to evaluate the interference of bioassay methods on the entomopathogenic fungi selection, using four Beauveria bassiana isolates and Alphitobius diaperinus adults. Regarding the inoculation form on insect mortality, the following methods were tested: insects immersion in a conidia suspension; spraying of a suspension over the insects; and surface treatment with eight concentrations. In order to evaluate the interference of immersion time on mortality, insects were immersed in a fungal suspension for 1, 10, 20, 30, 40, 50, and 60 seconds. Conidia produced on rice, insects, and synthetic medium were used to evaluate the influence of substrate on the virulence of the isolates. UNIOESTE 4 isolate and CG 152 were the most virulents for the methods immersion and spray, respectively; however, mortality was low in all isolates for the surface treatment. Mortality was proportional to the time during which insects stayed immersed, with a tendency to become stable after 40 seconds. Conidia from isolates CG 71 and CG 152 produced on insects were less virulent. The methods may influence bioassay results in the fungi selection for insects control.

O crescimento da produção avícola provocado pela alta demanda do mercado acarreta um aumento da criação de aves em confinamento, o que resulta em um aumento da densidade das aves dentro do galpão. Em função disso, há um incremento da umidade da cama aviária, proveniente tanto das excretas das aves, como da água dos bebedouros, que favorece o crescimento de populações do coleóptero Alphitobius diaperinus, popularmente conhecido como cascudinho. O contato direto do inseto com a cama das aves, assim como a alimentação a partir de aves moribundas e mortas, faz do A. diaperinus um veiculador de diversos patógenos, sendo destacados bactérias, protozoários e vírus que causam imunossupressão nas aves. Além da relação com os patógenos, os cascudinhos podem provocar danos nas instalações avícolas, pois, na fase larval, escavam túneis no material isolante para empuparem e destroem a proteção de poliuretano usada para isolamento térmico dos galpões em países de clima frio, a qual precisa ser trocada a cada dois ou três anos. Outro fator prejudicial às aves é a substituição da ração balanceada por larvas e adultos do coleóptero, uma vez que afeta o ganho de peso, principalmente, das mais jovens. Frente ao exposto, a presença do cascudinho em instalações avícolas torna-se um problema, tanto sanitário, como financeiro. O controle do A. diaperinus é considerado difícil, e inseticidas químicos são comumente utilizados em aviários, mas apresentam desvantagens, pois deixam resíduos na carcaça, bem como no meio ambiente. A presença de alta quantidade de matéria orgânica nos aviários provoca a diminuição da efetividade dos inseticidas químicos. Vários estudos vêm sendo desenvolvidos na busca de alternativas, como, por exemplo, nematoides e fungos entomapatogênicos, mas até o momento não são produzidos em escala comercial. Outra linha de pesquisa é a utilização de terra diatomácea, um pó-inerte que não deixa resíduos na carne de frango e no ambiente. The growth in poultry production because of the high market's demand causes an increase in the confinement raising, which increases the density of birds in poultry houses. As a result of this function, there is moisture increase in broiler litter, both from the excreta and drinkers, which favors the growth of the coleopterons Alphitobius diaperinus, popularly known as darkling beetle. The direct contact of the insects with the broiler litter as well as their feeding from dead and sick broilers make the A. diaperinus a vehicle for various pathogens, especially bacteria, protozoa and viruses. Besides the relationship with the pathogens, the darkling beetle can cause damage to poultry house, because at the larva stage they tent to burrow in to the insulating material and destroying the protection of polyurethane used for thermal insulation of poultry houses in countries with cold climate, which have to be changed every two or three years. Another harming factor to the broilers is the replacement of theirs balanced diet for larvae and adults of coleopterons, which affect the weight gain, mainly within younger individuals. Due to this factor, the presence of darkling beetle in poultry installations becomes a health as well as financial problem. The control of A. diaperinus is regarded as difficult and the use of chemical insecticides is common in poultry raising, even though it has disadvantages because it leaves residue in the carcass and in the environment. The presence of high amount of organic matter in poultry causes the decrease of the effectiveness of chemical insecticides. Several studies have been developed in research for alternatives, such as nematodes, fungi entomopathogenics, but they are not yet available commercially. Another line of research is the use of diatomaceous earth, an inert and non toxic powder which leaves no residues in chicken meat or in the environment.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:59:51Z No. of bitstreams: 1 S0103-84782007000300048.pdf: 31248 bytes, checksum: 5e209ebcbef777bd63142bb218c39023 (MD5) Made available in DSpace on 2013-08-22T18:59:51Z (GMT). No. of bitstreams: 1 S0103-84782007000300048.pdf: 31248 bytes, checksum: 5e209ebcbef777bd63142bb218c39023 (MD5) Previous issue date: 2007-06-01 Made available in DSpace on 2013-09-30T19:57:17Z (GMT). No. of bitstreams: 2 S0103-84782007000300048.pdf: 31248 bytes, checksum: 5e209ebcbef777bd63142bb218c39023 (MD5) S0103-84782007000300048.pdf.txt: 14731 bytes, checksum: fa5ba51ad4a9c08df65395aabe2dc5c6 (MD5) Previous issue date: 2007-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:15:16Z No. of bitstreams: 2 S0103-84782007000300048.pdf: 31248 bytes, checksum: 5e209ebcbef777bd63142bb218c39023 (MD5) S0103-84782007000300048.pdf.txt: 14731 bytes, checksum: fa5ba51ad4a9c08df65395aabe2dc5c6 (MD5) Made available in DSpace on 2014-05-20T13:15:16Z (GMT). No. of bitstreams: 2 S0103-84782007000300048.pdf: 31248 bytes, checksum: 5e209ebcbef777bd63142bb218c39023 (MD5) S0103-84782007000300048.pdf.txt: 14731 bytes, checksum: fa5ba51ad4a9c08df65395aabe2dc5c6 (MD5) Previous issue date: 2007-06-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) O besouro Alphitobius diaperinus spp (cascudinho) é visto como uma importante praga da avicultura mundial. Por suas características comportamentais e hábitos biológicos que dificultam seu controle, é considerado um vetor de agentes patogênicos. O objetivo desta pesquisa foi investigar o cascudinho como possível vetor de Clostridium perfringens em granjas avícolas industriais, localizadas em diferentes regiões do interior Paulista. Através de métodos bacteriológicos convencionais, em 40 amostras analisadas, foram encontradas contagens significativas de Clostridium perfringens em todas elas. A partir dos resultados obtidos, pôde-se demonstrar o potencial deste inseto como vetor do agente responsável pela enterite necrótica. The Alphitobius diaperinus spp (lesser mealworm) is considered an important world poultry plague. Due to its behavior characteristics and biological habits that make its control difficult it is considered a vector of pathogenic agents. The objective of this research was to investigate the little mealworm as possible vector of Clostridium perfringens in broiler houses, located in different parts of the state of São Paulo. Through conventional bacteriological methods, 40 samples of little mealworm collected were analyzed. Clostridium perfringens was found in all of the samples and the potential of this insect as vector of the necrotic enteritis was demonstrated. Universidade Estadual Paulista Departamento de Microbiologia Universidade Estadual Paulista Departamento Medicina Veterinária Preventiva Universidade Estadual Paulista Departamento de Microbiologia Universidade Estadual Paulista Departamento Medicina Veterinária Preventiva

Knowledge of the population fluctuation and spatial distribution of pests is fundamental for establishing an appropriate control method. The population fluctuation and spatial distribution of the Alphitobius diaperinus in a poultry house in Cascavel, in the state of Parana, Brazil, was studied between October, 2001 and October 2002. Larvae and adults of the lesser mealworm were sampled weekly using Arends tube traps (n = 22) for six consecutive flock grow-outs. The temperature of the litter and of the poultry house was measured at the same locations of the tube traps. Beetle numbers increased continuously throughout all the sampling dates (average 5,137 in the first week and 18,494 insects on the sixth week). Significantly greater numbers of larvae were collected than adults (1 to 20 times in 95% of the sampling points). There was no correlation between temperature and the number of larvae and adults collected, therefore no fluctuation was observed during the sampling period. The population growth was correlated to litter re-use. The highest temperatures were observed in deep litter. The spatial distribution of larvae and adults in the poultry house was heterogeneous during the whole period of evaluation. Results suggest that monitoring in poultry houses is necessary prior to adopting and evaluating control measures due to the great variability of the insect distribution in the poultry house. O conhecimento da flutuação populacional e distribuição espacial de pragas são fundamentais para o estabelecimento de uma metodologia de controle adequada. A flutuação populacional e distribuição espacial de Alphitobius -diaperinus em aviário de corte, localizado em Cascavel, Paraná foi avaliada no período entre outubro/2001 e Outubro/2002. Larvas e adultos do cascudinho foram coletados semanalmente com armadilhas de Arends (n = 22) em seis lotes de frangos, consecutivamente. A temperatura da cama foi medida nos locais onde as armadilhas foram expostas, bem como a temperatura no interior do galpão do aviário. O número de besouros aumentou continuamente em todas as áreas do aviário no decorrer das semanas de coleta (média de 5.137, na primeira semana, e de 18.494 insetos, na sexta semana). O número de larvas coletadas foi significantemente maior que o número de adultos (de 1 a 20 vezes em 95% do total de coletas realizadas). Não houve correlação entre as variações de temperatura do galpão e da cama e o número de insetos coletados, não sendo, portanto, observada flutuação populacional ao longo do período de avaliação. O crescimento da população esteve relacionado ao acúmulo de camas, observando-se temperaturas mais altas em locais onde a cama era mais profunda. A distribuição espacial de larvas e adultos no galpão foi desuniforme com relação aos locais de coleta. Com base nos resultados obtidos, sugere-se que há necessidade de monitoramento dos aviários individualmente antes do emprego de qualquer medida de controle, devido à grande variabilidade na distribuição dos insetos em um único galpão de criação.