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In vitro assay and morphological characterization of a new product based on diatomaceous earth for lesser mealworm control in poultry houses

Avaliação in vitro e caracterização morfológica de um novo produto à base de terra de diatomáceas visando ao controle do cascudinho dos aviários

ABSTRACT:

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%).

KEYWORDS:
lesser mealworm; alternative control; inert powder

RESUMO:

Terra de diatomáceas (TD) é um pó inerte proveniente da moagem de rochas de depósitos de algas diatomáceas fossilizadas rico em dióxido de sílica. Apresenta-se como um pó fino e altamente higroscópico e é recomendado para uso em aviários de frango de corte como condicionador ambiental, com ação secundária na redução populacional do cascudinho (Alphitobius diaperinus). Este estudo visou avaliar, em laboratório, a atividade inseticida de um novo produto comercializado no Brasil como condicionador ambiental de aviários (Poultry Sec®, Vet Science Bio Solution, Maringá, Paraná, Brasil), bem como caracterizá-lo. O experimento foi conduzido adicionando-se o produto na ração de frango (1, 2 and 3 g TD/kg ração), e também com a aplicação dele na cama de aviário (86 e 172 g TD/m2). Os insetos adultos foram mantidos nos recipientes com as misturas em 26°C no escuro por 10 dias. Amostras do produto foram também analisadas em microscópio óptico visando à contagem e medição das frústulas e identificação das espécies presentes no produto. Quando aplicado sobre a cama, o produto causou entre 83 e 85% de mortalidade. Foram estimados 1,2 × 106 indivíduos/g do produto, distribuídos em nove gêneros diferentes, com predominância de Brachysira Kützing (40%).

PALAVRAS-CHAVE:
cascudinho dos aviários; controle alternativo; pó inerte

INTRODUCTION

The lesser mealworm (Alphitobius diaperinus) is a worldwide pest of poultry facilities. During its life cycle, larvae live in poultry litter and migrate to the soil for pupation. Adults live in the litter (CHERNAKI; ALMEIDA, 2001CHERNAKI, A.M.; ALMEIDA, L.M. Exigências térmicas, período de desenvolvimento e sobrevivência de imaturos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.30, p.365-368, 2001.). These insects are ingested by the poultry instead of feed, which affects the growth and health of the birds because these insects are also a potential vector of pathogenic bacteria such as Salmonella sp., Campylobacter sp. and Clostridium sp. (SKOV et al., 2004SKOV, M.N.; SPENCER, A.G.; HALD, B.; NAUERBY, B.; CARSTENSEN, B.; MADSEN, M. The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler floks. Avian Diseases, Jacksonville, v.48, p.9-18, 2004.; VITTORI et al., 2007VITTORI, J.; SCHOCKEN-ITURRINO, R.P.; TROVÓ, K.P.; RIBEIRO, C.A.M.; BARBOSA, G.G.; SOUZA, L.M.; PIGATTO, C.P. Alphitobius diaperinus como veiculador de Clostridium perfingens em granjas avícolas do interior paulista - Brasil. Ciência Rural, Santa Maria, v.37, p.894-896, 2007.; HAZELEGER et al., 2008HAZELEGER, W.C.; BOLDER, N.M.; BEUMER, R.R.; JACOBS-REITSMA, W.F. Darkling Beetles (Alphitobius diaperinus) and Their Larvae as Potential Vectors for the Transfer of Campylobacter jejuni and Salmonella enterica Serovar Paratyphi B Variant Java between Successive Broiler Flocks. Applied and Environmental Microbiology, Washington, v.74, p.6887-6891, 2008.; JAPP et al., 2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.; CHERNAKI-LEFFER et al., 2010CHERNAKI-LEFFER, A.M.; KUTTEL, J.; MARTINS, L.M.; PEDROSO, A.C.; ASTOLFI-FERREIRA, C.S.; FERREIRA, F.; PIANTINO-FERREIRA, A.J. Vectorial competence of larvae and adults of Alphitobius diaperinus in the transmission of Salmonella Enteritidis in poultry. Vector-Borne and Zoonotic Diseases, New Rochelle, v.10, p.481-487, 2010.).

This lesser mealworm is controlled by pyrethroid insecticides applied superficially on the poultry litter. In addition to their inefficiency in insect control, environmental and bird health risks are observed with these insecticide applications. Additionally, the emergence of resistant insect populations can occur (GARG et al., 2004GARG, U.K.; PAL A.K.; JHA, G.J.; JADHAO, S.B. Pathophysiological effects of chronic toxicity with synthetic pyrethroid, organophosphate and chlorinated pesticides on bone health of broiler chicks. Toxicologic Pathology, Research Triangle Park, v.32, p.364-369, 2004.; UEMURA et al., 2008UEMURA, D.H.; ALVES, L.F.A.; OPAZO, M.A.U.; ALEXANDRE, T.M.; OLIVEIRA, D.G.P.; URSI, M.V. Distribuição e dinâmica populacional do cascudinho Alphitobius diaperinus (Coleoptera: Tenebrionidae) em aviários de frango de corte. Arquivos do Instituto Biológico, São Paulo, v.75, p.429-435, 2008.; CHERNAKI et al., 2011CHERNAKI, A.M.; SOSA-GOMEZ, D.R.; ALMEIDA, L.M.; LOPES, I.O.N. Suscetibility of Alphitobius diaperinus to cypermethrin, dichlorvos and triflumuron in Southern Brazil. Revista Brasileira de Entomologia, Curitiba, v.55, p.125-128, 2011.). Studies with entomopathogenic fungi (ROHDE et al., 2006ROHDE, C.; ALVES, L.F.A.; NEVES, P.M.O.J.; ALVES, S.B.; SILVA, E.R.L.; ALMEIDA, J.E.M. Seleção de isolados de Beauveria bassiana (Bals.) Vuill. e Metarhizium anisopliae (Metsch.) Sorok. contra o cascudinho Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.35, p.231-240, 2006.; ALVES et al., 2015ALVES, L.F.A.; OLIVEIRA, D.G.P.; LAMBKIN, T.; BONINI, A.K.I.; ALVES, V.M.; PINTO, F.G.S.; SCUR, M.C. Beauveria bassiana applied to broiler chicken houses as biocontrol of Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae), an avian pathogens vector. Brazilian Journal of Poultry Science, Campinas, v.17, p.459-466, 2015.), insecticidal plants (MARCOMINI et al., 2009MARCOMINI, A.M.; ALVES, L.F.A.; BONINI, A.K.; MERTZ, N.R.; SANTOS, J.C. Atividade inseticida de extratos vegetais e do óleo de neem sobre Adultos de Alphitobius diaperinus Panzer (Coleoptera, Tenebrionidae). Arquivos do Instituto Biológico, São Paulo, v.76, p.413-420, 2009.; ALVES et al., 2012ALVES, V.M.; ALVES, L.F.A.; UEMURA-LIMA, D.H. Atividade da torta de nim sobre adultos do cascudinho dos aviários em condições de laboratório. Ciência Rural, Santa Maria, v.42, p.888-893, 2012.) and inert powders (SANTORO et al., 2010SANTORO, P.H.; NEVES, P.M.O.J.; AMARO, J.T.; CONSTANSKI, K.; LÓPEZ, E.A.L.; ALVES, L.F.A. Associação de pós inertes com fungo entomopatogênico para o controle do cascudinho (Alphitobius diaperinus). Ciência Rural, Santa Maria, v.40, p.1354-1359, 2010.; ALVES et al., 2012ALVES, V.M.; ALVES, L.F.A.; UEMURA-LIMA, D.H. Atividade da torta de nim sobre adultos do cascudinho dos aviários em condições de laboratório. Ciência Rural, Santa Maria, v.42, p.888-893, 2012.) have shown them to be potential alternatives for insect control. Among the latter, highlight is given to diatomaceous earth (DE). It is non-toxic to birds and is used for the control of stored grain pests. The insecticidal action of DE against the lesser mealworm has been proven in the laboratory (ALVES et al., 2006ALVES, L.F.A.; BUSARELLO, G.D.; OLIVEIRA, D.G.P.; ALVES, S.B. Ação da terra de diatomácea contra adultos do cascudinho Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae). Arquivos do Instituto Biológico, São Paulo, v.73, p.115-118, 2006.; JAPP et al., 2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.; JAPP et al., 2010JAPP, A.K.; BICHO, C.L.; SILVA, A.V.F. Importância e medidas de controle para Alphitobius diaperinus em aviários. Ciência Rural, Santa Maria, v.40, p.1668-1673, 2010.).

DE is highly hygroscopic and it has been recommended as a way to lower the water activity and humidity of poultry litter and to reduce the formation of ammonia, as already demonstrated with other powders that are mixed into the poultry litter (WATSON et al., 2003WATSON, D.W.; DENNING, S.S.; ZUREK, L.; STRINGHAM, S.M.; ELLIOTT, J. Effects of lime hydrate on the growth and development of darkling beetle, Alphitobius diaperinus. International Journal of Poultry Science, Faisalabad, v.2, p.91-96, 2003.). At the same time, DE acts on the lesser mealworm by causing abrasions on the protective wax coating on its cuticle, leading to the loss of water from the insect’s body, resulting in death (EBLING, 1971; KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.). Moreover, our previous studies in the laboratory and in a commercial poultry house corroborated by results obtained by LAMBKIN et al. (2012LAMBKIN, T.A.; SWAIN, A.J.; RICE, S.J.; BARTLETT, J.S.; ZALUCKI, M.P. Field Assessments of Control Agents for Lesser Mealworm (Coleoptera: Tenebrionidae) Using Litter Sampling. Journal of Economic Entomology, Riverside, v.105, p.979-987, 2012.), point to the benefits of using DE in poultry houses.

DE is made of fossilized diatomaceous algae, composed of amorphous silicon dioxide (YANG et al., 1998YANG, H.S.; LEE, W.J.; CHEN, S.J.; LAI, S.O. PAH emission from various industrial stacks. Journal of Hazardous Materials, Buffalo, v.60, p.159-174, 1998.; WU et al., 2005WU, J.; YANG, Y.S.; LIN, J. Advanced tertiary treatment of municipal wastewater using raw and modified diatomite. Journal of Hazardous Materials, Buffalo, v. 127, p.196-203, 2005.) with a wide variety of species, morphologies (centrics, penates etc.) and chemical compositions (ROUND et al., 1992ROUND, F.E.; CRAWFORD, R.M.; MANN, D.G. The diatoms. Biology and morphology of the genera. New York: Cambridge University Press, 1992. 747p.; KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.).

During its processing, DE is submitted to a calcination process that reduces its moisture, organic material, frustule surface area and density (DOLLEY, 2000DOLLEY, T.P. Diatomite. U.S. Geological Survey Minerals Yearbook, v.25, p.1-6, 2000.). The latter is very important in DE efficiency, being inversely proportional to the hygroscopicity (WU et al., 2005WU, J.; YANG, Y.S.; LIN, J. Advanced tertiary treatment of municipal wastewater using raw and modified diatomite. Journal of Hazardous Materials, Buffalo, v. 127, p.196-203, 2005.). The hygroscopicity of DE is also due to the very small inner pores of the frustules (EBLING, 1971).

In addition to the hygroscopicity, the DE efficacy is subject to the abrasivity given to the presence of silicon dioxide (KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.; KHRAISHEH et al., 2004KHRAISHEH, M.A.M.; AL-DEGS, Y.S.; MCMINN, W.A.M. Remediation of wastewater containing heavy metals using raw and modified diatomite. Chemical Engineering Journal, Cambridge, v.99, p.177-184, 2004.; WU et al., 2005WU, J.; YANG, Y.S.; LIN, J. Advanced tertiary treatment of municipal wastewater using raw and modified diatomite. Journal of Hazardous Materials, Buffalo, v. 127, p.196-203, 2005.), the morphology of the predominant frustules and by particle sizes (SOUZA, 1973SOUZA, J.F. Perfil analítico da diatomita. Rio de Janeiro: Departamento Nacional da Produção Mineral, 1973. 27p. (Boletim 11).).

A previous study of different natural samples of DE proved that the physical characteristics of DE, including mineral origin, mineral composition and particles size affect its efficacy in the control of lesser mealworm (JAPP, 2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.). In addition, KORUNIC (1997KORUNIC, Z. Rapid assessment of the insecticidal value of diatomaceous earths without conducting bioassays. Journal of Stored Products Research, Manhattan, v.33, p.219-229, 1997.) noted the importance of knowing about the physical and chemical properties of DE as a tool to help experts evaluate the quality of a DE product. However, nothing is known about characterization of insecticides based on DE in Brazil (most of which are registered for lesser mealworm control). So, it is clear that there is a need for more information about the activity and characterization of DE products and this pioneer study was carried out to meet this objective.

Experiments were conducted in the laboratory to assess the insecticidal action of the product Poultry Sec® (Vet Science Bio Solution, Maringá, Paraná, Brazil, www.vetscience.com.br) on the lesser mealworm adults to determine the optimal concentration for insecticidal activity and check its performance as a component in the poultry litter.

Additionally, a microscopic analysis was performed in order to check the size, morphology and density of the skeleton of diatom density.

MATERIAL AND METHODS

In all bioassays, insects were collected in a commercial poultry house and maintained in the laboratory (26 ± 1ºC, 12 h photoperiod) in sealed plastic containers with perforated lids, with either poultry litter or poultry feed.

Initially, we performed tests to determine the optimal concentration of Poultry Sec®. The product was evaluated by two different strategies: mixed into poultry feed and mixed into poultry litter. Firstly, Poultry Sec® was mixed into poultry feed (dried previously at 40°C, 24 h) in polyethylene bags under constant stirring for two minutes at doses equivalent to 1, 2 and 3 g of the product/kg of poultry feed. After that, the mixture was placed in 10 cm diameter plastic containers (4 cm depth) and then 30 insects were released in each container (ALVES et al., 2006ALVES, L.F.A.; BUSARELLO, G.D.; OLIVEIRA, D.G.P.; ALVES, S.B. Ação da terra de diatomácea contra adultos do cascudinho Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae). Arquivos do Instituto Biológico, São Paulo, v.73, p.115-118, 2006.). We prepared four repetitions for each concentration. Insects were kept on pure poultry feed in control. For all treatments, insects were kept at 26 ± 1ºC, with a 12 h photoperiod for 10 days. After that, the dead insects were counted. This experiment was repeated twice.

For the poultry litter experiment, Poultry Sec® was mixed into the poultry litter at concentrations equivalent to 86 and 172 g/m2 and distributed in containers as previously described. After that, insects were transferred to the containers and 5 g of poultry feed was added. This experiment was repeated twice.

Normal distribution of the mortality data was checked by Shapiro-Wilk test. Data were analyzed by one-way ANOVA and Tukey’s test (p < 0.05) (FERREIRA, 2011FERREIRA, D.F. Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia, Lavras, v.35, p.1039-1042, 2011.).

For genus identification and size determination of the diatomaceous frustules (whole or fragmented), permanent slides were prepared and analyzed in a photomicroscope. Digital images were obtained. Identification was made based on KRAMMER; LANGE-BERTALOT (1986KRAMMER, K.; LANGE-BERTALOT, H. (Eds.). Naviculaceae: neue und wenig bekannte Taxa neue Kombinationen und Synonyme sowie Bemerkungen zu einigen Gattugen. Bibliotheca Diatomologica. Berlin: Koeltz Scientific Books, 1986. 229p.; 1991KRAMMER, K.; LANGE-BERTALOT, H. (Eds.). Bacillariophyceae: Centrales, Fragilariaceae, Eunotiaceae. Süsswasser flora von Mittleleuropa. Berlin: Koeltz Scientific Books , 1991. 576p.; 1992KRAMMER, K.; LANGE-BERTALOT, H. (Eds.). Pinnularia: eine Monographie der europäischen. Bibliotheca Diatomologica Taxa I. Berlin: Koeltz Scientific Books , 1992. 353p.), KRAMMER (1997aKRAMMER, K. (Ed.). Die cymbelloiden Diatomeen: eine monographie der weltweit bekannten. Bibliotheca Diatomologica Taxa I. Berlin: Koeltz Scientific Books , 1997a. 382p.; 1997bKRAMMER, K. (Ed.). Die cymbelloiden Diatomeen: eine monographie der weltweit bekannten. Bibliotheca Diatomologica Taxa II. Berlin: Koeltz Scientific Books , 1997b. 469p.), METZELTIN; LANGE-BERTALOT (1998METZELTIN, D.; LANGE-BERTALOT, H. (Eds.). Tropical Diatoms of South America I. Iconographia Diatomologica. Germany: Koeltz Scientific Books, 1998. 220p.; 2007METZELTIN, D.; LANGE-BERTALOT, H. (Ed.). Tropical diatoms of South America II. Iconographia Diatomologica. Germany: Koeltz Scientific Books , 2007. 320p.); METZELTIN et al. (2005METZELTIN, D.; LANGE-BERTALOT, H.; GARCÍA-RODRIGUES, F. (Eds.). Diatoms of Uruguay. Iconographia Diatomologica. Germany: Koeltz Scientific Books , 2005. 736p.); and ROUND et al. (1992ROUND, F.E.; CRAWFORD, R.M.; MANN, D.G. The diatoms. Biology and morphology of the genera. New York: Cambridge University Press, 1992. 747p.). Images were taken and analyzed with AxioVision 4.8 software.

Density was determined using the Uthermöl sedimentation method. Consequently, 0.0025 g DE samples were added to 10 mL of distilled water in a sedimentation chamber, as described by UTERMÖHL (1958UTERMÖHL, H. Zur Vervollkommung der Quantitativen Phytoplankton Methodik. Internationale Vereinigung für Theoretische und Angewandte Limnologie, Mitteilungen, v.9, p.1-38, 1958.); MARGALEF (1983MARGALEF, R. Limnologia. Barcelona: Omega; 1983. 1010p.); and BICUDO (1990BICUDO, D.C. Considerações sobre metodologias de contagem de algas do perifíton. Acta Limnologica Brasiliensia, São Paulo, v.3, p.459-475, 1990.). Density (individuals/mL) was calculated as APHA (1995APHA. AMERICAN PUBLIC ASSOCIATION. Standart methods for the examination of water and wasterwater. 19. ed. Washington, D.C.: APHA, 1995.): D = C (AT/AF×F×V), in which D = total density (organisms/mL); C = number of organisms; AT = total area of the sedimentation chamber bottom (mm2); AF = surface area of the field of counting (mm2); F = field counted number; V = sample volume (mL).

RESULTS AND DISCUSSION

The efficiency of the Poultry Sec® was observed, and mortality ranged from 50% (with the lowest concentration) to 80-90% (with the two highest concentrations) (Table 1).

Table 1:
Mortality (%) of lesser mealworm adults that had contact with different dosages of DE in the two application strategies (poultry feed and poultry litter), 10 days after treatment (26ºC, 12 h photophase).

Similar results indicating the high insecticidal activity of a commercial product based on DE against the lesser mealworm have already been obtained (mortality above 80%) (ALVES et al., 2006ALVES, L.F.A.; BUSARELLO, G.D.; OLIVEIRA, D.G.P.; ALVES, S.B. Ação da terra de diatomácea contra adultos do cascudinho Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae). Arquivos do Instituto Biológico, São Paulo, v.73, p.115-118, 2006.; 2008ALVES, L.F.A.; OLIVEIRA, D.G.P.; NEVES, P.M.O.J. Fatores que afetam a eficiência da Terra de Diatomácea no controle de adultos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.37, p.716-722, 2008.). Also, samples of DE from different Argentine regions were applied directly to lesser mealworm (63 g/m2) causing from 7 to 98% of mortality (JAPP, 2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.).

Although the variations on insecticidal activity may be explained by the morphological or chemical characteristics of the poultry litter (ALVES et al., 2008ALVES, L.F.A.; OLIVEIRA, D.G.P.; NEVES, P.M.O.J. Fatores que afetam a eficiência da Terra de Diatomácea no controle de adultos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.37, p.716-722, 2008.; OLIVEIRA et al., 2009OLIVEIRA, D.G.P.; ALVES, L.F.A.; MARCHESE, L.P.C.; THOMAZONI, D. Persistência da ação inseticida e repelência da Terra de Diatomácea para cascudinho de aviário Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae). Semina, Londrina, v.30, p.201-210, 2009.), the differences between the techniques used here and by the JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.) experiments are notable.

While the products were applied directly to insects by JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.), in this study, Poultry Sec® was mixed with poultry feed or poultry litter. This approach simulated field conditions but strongly reduced the contact between the product and the insects. However, the Poultry Sec® concentration was 20 to 40% greater than that used by JAPP (2008). Additionally, ALVES et al. (2008ALVES, L.F.A.; OLIVEIRA, D.G.P.; NEVES, P.M.O.J. Fatores que afetam a eficiência da Terra de Diatomácea no controle de adultos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.37, p.716-722, 2008.) proved by microscope electronic analysis that the wood shavings from poultry litter reduce the transference of DE to insects in comparison to a poultry feed substrate.

DE is a natural product that is formed from fossilized diatomaceous algae of various shapes and sizes. This is reflected in the different physical characteristics between DE samples from different natural deposits, and it also explains the variations of insecticidal activity of different DE products (KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.).

Poultry Sec® was observed to contain particles with frustules of 3 to approximately 50 µm, and a few of 120 µm. These sizes are close to the medium particle size found in different commercial DE products, reported by KORUNIC (1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.). On the other hand, JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.) analyzed DE samples from different mines in Argentina, and observed larger particles (500 to 3,000 µm) than we observed in Poultry Sec®. As noted by KORUNIC (1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.), particles up to 10 µm and higher amounts of amorphous silica are essential for high insecticidal activity, what explains the results obtained with Poultry Sec®.

The natural origin of DE also explains the differences in its chemical composition, because the amorphous silica content in DE varies from 60 to 90% (KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.). This difference is observed in the tests of DE samples containing 60 to 75% silicon and the Poultry Sec® used in this study, which, according to the manufacturer, contains 86% silicon. In this sense, microscopic analysis proved that Poultry Sec® has a higher silicon concentration based on the counting of 1.2 × 106 diatom frustules/g from very rich silicon diatom species (KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.). However, KORUNIC (1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.) affirms that it is not possible to bind the insecticidal action of DE to silicon concentrations. In this way, JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.) found that DE samples that were high in silicon were not the most efficient ones. The opposite effect has also been observed.

Thus, it is concluded that the most important factor to determine the activity of DE is the morphology of the frustules, proving the importance of this study.

Regarding the diatom species that make up Poultry Sec®, 322 frustules from nine different genera were found, with eight belonging to the Bacillariophyceae class and one to the Fragilariophyceae class (pennates frustules). Additionally, the genus Brachysira Kützing was the most prevalent one (40%) (Table 2, Fig. 1). Contrary to the results reported by JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.), Coscinodiscophyceae species (centrics frustules) were not found.

Table 2:
Diatom genera present in Poultry Sec® (× 40 magnification).

Figure 1:
Diatom genera observed in the sample of Poultry Sec®: 1-5) Brachysira Kützing; 6-10) Eunotia Ehrenberg; 11-12) Encyonema Kützing; 13-15) Nitzschia Hassall; 16-18) Staurosirella Williams & Round; 19-22) Frustulia Agardh; 23-24) Pinnularia Ehrenberg; 25-27) Navicula Bory; 28-30) Actinella Lewis (× 1000).

As previously discussed, the technique used to mix the Poultry Sec® into the poultry litter leads to dispersion, and scanning electron microscopy proved that wood shaving poultry litter is more efficient than poultry feed in removing DE from the lesser mealworm body surface (ALVES et al., 2008ALVES, L.F.A.; OLIVEIRA, D.G.P.; NEVES, P.M.O.J. Fatores que afetam a eficiência da Terra de Diatomácea no controle de adultos de Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Neotropical Entomology, Londrina, v.37, p.716-722, 2008.). Although DE was more dispersed in the litter substrate, the insecticide efficiency was not affected.

Thus, the efficiency of Poultry Sec® is probably due to the uniformity and small size of the particles (3-50 µm), which is essential to give Poultry Sec® mobility in poultry litter wood shavings. In this way, DE is physically stable and will affect insects as long as it is dry and in sufficient concentration to ensure that the insects come in contact with enough diatom particles (KORUNIC, 1998KORUNIC, Z. Diatomaceous earths, a group of natural insecticides. Journal of Stored Products Research, Manhattan, v.34, p.87-97, 1998.).

Although there are several products based on DE recommended for insect control, there are very few products for lesser mealworm control. JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.) and the results from this study indicate that the diatomaceous composition changes between products from different parts of the word. In Brazil, the total lack of technical information available on commercialized products based on DE makes comparisons difficult. The data obtained here and corroborated by JAPP (2008JAPP, A.K.; HUBER, R.M.; SANTOS, S.; MEURER, R.; BICHO C.L.; SILVA, A.V.F. Interferência no ganho de peso de frangos de corte alimentados com cascudinhos Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae). Revista Brasileira de Ciência Avícola, Campinas, v.10, p.170, 2008.) highlight the importance of the physical characterization as one of the main reasons for the variations in tests results, regarding the efficiency of DE-based products.

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Publication Dates

  • Publication in this collection
    2017

History

  • Received
    24 Aug 2015
  • Accepted
    16 Mar 2017
Instituto Biológico Av. Conselheiro Rodrigues Alves, 1252 - Vila Mariana - São Paulo - SP, 04014-002 - São Paulo - SP - Brazil
E-mail: arquivos@biologico.sp.gov.br