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Diante do aumento da população mundial, consumo em excesso e consequente poluição ambiental, a busca por alternativas eficientes que contribuam para a descontaminação das águas é de grande relevância. Os efluentes industriais apresentam composição física, química e biológica complexa, sendo que o tipo de matéria prima usada pela indústria definirá sua aparência, coloração, densidade, acidez e basicidade, temperatura e presença de compostos orgânicos e/ou inorgânicos. O descarte incorreto desse passivo ambiental pode causar danos à saúde e prejuízos ao ecossistema. Nesse contexto, a utilização da tecnologia de Wetlands Construídas (WC) para o tratamento de águas residuais industriais tem se tornado uma perspectiva crescente, visto suas inúmeras vantagens face aos tratamentos convencionais, como baixo custo, simplicidade na implantação e alta eficiência na remoção de poluentes emergentes. WC é um mecanismo, composto por lagoas ou canais artificiais rasos, que abrigam plantas aquáticas que possuem um leito filtrante, onde essas espécies vegetais promovem a atenuação de diversos contaminantes presentes no efluente. Assim, o objetivo deste trabalho é realizar uma revisão bibliográfica, preferencialmente no período de (2011/2021), sobre o uso de WC na fitorremediação de efluentes industriais, que ainda é uma alternativa pouco difundida no contexto nacional. Nesta revisão, diversas modalidades de WC que apresentam alto grau de eficiência para remoção de compostos poluentes dos efluentes industriais foram encontradas. Para além do mais, pode-se concluir que a aplicação dessa tecnologia representa a possibilidade de expandir o tratamento descentralizado de efluentes, alcançando diversas localidades de países em desenvolvimento, e também reduzir as emissões de gases de efeito estufa, gerados pelos tratamentos anaeróbios.

A preocupação das populações de vários países e da comunidade científica global tem sido crescente nos últimos anos com relação às mudanças climáticas. O Aquecimento Global atinge níveis alarmantes e gera apreensão na comunidade internacional que teme danos irreversíveis ao planeta. A intensificação do Efeito Estufa em virtude da emissão excessiva de gases como o dióxido de carbono tem agravado esse cenário. Frente a esse problema, muitos pesquisadores têm dedicado seus esforços para criar tecnologias que visam neutralizar ou compensar os danos gerados pelos gases poluentes. Entre as alternativas disponíveis, o biossequestro de carbono ganhou espaço nos últimos anos, seu objetivo é utilizar organismos vivos para aprisionar o carbono atmosférico de forma a minimizar os impactos do aumento da temperatura do planeta. Esse artigo apresenta uma revisão da utilização de Escherichia coli, Microalgas e Árvores Geneticamente Modificadas, como biossequestradores potenciais de carbono atmosférico.

Planococcus citri (Risso 1813) (cochonilha-branca) é uma praga de plantas do gênero Citrus. Quando ocorre em grande quantidade, causa desfolhas nas plantas e queda de frutos. Esta pesquisa visou desenvolver uma atividade extracurricular com alunos do primeiro ano do ensino médio (escola estadual), para realização de um levantamento qualitativo de informações acerca do controle entomológico alternativo na área urbana da cidade de Rubim (MG), focando os biocidas para Planococcus citri. Trata-se de um estudo qualitativo e descritivo. No embasamento teórico, foram utilizados autores que estudam P. citri e que trazem concepções relacionadas aos processos de ensino e de aprendizagem. As atividades desenvolvidas consistiram em observações in natura, em laboratório, discussões, entrevistas, registros fotográficos e anotações relacionadas ao inseto. Os resultados apontam que a população da cidade de Rubim utiliza estratégias alternativas para o controle do inseto em questão. As análises e discussões realizadas pelos alunos contribuíram para o ensino e a aprendizagem de conceitos relacionados com conteúdos de Biologia e com a realidade cotidiana. Os estudantes sentiram-se estimulados, promoveram envolvimento e interações entre si e com a comunidade. Ressalta-se que os estudos em que são empregados métodos diferenciados, interagindo com recursos tecnológicos, possibilitam um aprendizado mais eficaz.

Sitophilus zeamais Motschulsky, 1855 (Coleoptera: Curculionidae) which develops inside the grain, is an aggressive pest of stored grains. Here, we evaluated the efficiency of diatomaceous earth (DE), Diatomita CI/325 CX, and dolomite (DOL), Dolomita Dr. Kanyo, against S. zeamais reared on corn under laboratory conditions and observed external alterations on treated insects using scanning electron microscopy (SEM). Insects treated with both DE and DOL were impregnated with particles on the body surface, and they showed 100% mortality after 10 days of exposure. Damages caused by inert dusts may have contributed to dehydration and desiccation, consequently, leading insects to death. Thus, diatomaceous and dolomite were effective against S. zeamais under laboratory conditions.

The Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) has a worldwide distribution and is considered of medical and veterinary importance. Its synanthropic behavior combined with the visitation of decaying organic matter allows it to mechanically transmit pathogens to human and animals. Pyrethroids are widely used pesticides, effective in low doses against a broad spectrum of insects. Our objective was to evaluate the action of K-Othrine® on the post-embryonic development of C. megacephala. First instar larvae were exposed to different concentrations of K-Othrine® (3.75 × 10−5 g a.i/mL (low), 7.5 × 10−5 g a.i/mL (intermediate low), 1.125 × 10−4 g a.i/mL (intermediate), 1.5 × 10−4 g a.i/mL (intermediate high), and 2.5 × 10−4 g a.i/mL (high)) mixed into artificial diets and a control with no pesticide. We measured weight, time of development, sex ratio, larval and pupal mortality, and esterase patterns. The mean weight values at intermediate low and intermediate concentrations were significantly lower compared with the control; additionally, the development time was delayed compared with the control. The larval and pupal mortality differed among the treatments. The larvae had six esterase regions, which altered its relative activity at 48 h and 72 h after insecticide ingestion. Further studies must investigate the effects of different insecticides on the post-embryonic development of C. megacephala.

Os insetos apresentam a maior biodiversidade entre a classe Animal e são fontes ricas de produtos biotecnológicos. Espécies da família Calliphoridae (Diptera) apresentam hábitos necrófagos durante a fase larval, assim são expostas a diferentes microrganismos, produzindo peptídeos antimicrobianos (AMPs) como sistema de defesa. O objetivo deste trabalho foi realizar um levantamento bibliográfico dos principais peptídeos antimicrobianos identificados em Calliphoridae (Diptera), fornecendo dados para a bioprospecção de novos compostos. A revisão bibliográfica foi realizada nas plataformas: Google Acadêmico e SciFinder e foram selecionados 56 artigos para o desenvolvimento deste trabalho. Os AMPs possuem atividade contra amplo espectro de bactérias gram-positivas e gram-negativas, por isso apresentam grande interesse para tratamentos de feridas (e.g.: Terapia Larval) e bioprospecção de novos antibióticos. Estas moléculas são obtidas a partir do produto de excreção e secreção das larvas de Calliphoridae e sua ação tem como alvo primário a membrana celular das bactérias, devido interações eletroestáticas e interações de van der Waals com a membrana lipídica. A utilização de técnicas de separação acopladas com espectrometria de massas permitiu a identificação e caracterização de peptídeos da classe Defensina em diferentes espécies de califorídeos. A bioprospecção de novas moléculas antimicrobianas em moscas é uma área promissora dentro da biotecnologia, devido ao aumento do número de bactérias resistentes aos antibióticos atuais.

BACKGROUND Bacillus thuringiensis (Bt) is a Gram-positive bacterium that synthesizes specific protein toxins, which can be exploited for control of various insect pests, including Diatraea saccharalis, a lepidopteran that severely damages sugarcane crops. Although studies have described the effects of Bt in the larval phases of D. saccharalis, few have examined its effect on insect eggs. Herein, we studied the entomopathogenic potential of Bacillus thuringiensis serovar Aizawai GC-91 (Bta) during D. saccharalis embryo development with the aim of understanding the entomopathogenic mechanism and developing new biological control techniques for target insects. RESULTS Bta concentrations of 5, 10 and 20 g L-1 demonstrated the strongest bioactivity, reducing D. saccharalis egg viability by 28.69%, 33.91% and 34.98%, respectively. The lethal concentrations (LCs) were estimated as: LC50 = 28.07 g L-1 (CI 95% = 1.89-2.38) and LC90 = 65.36 g L-1 (CI 95% = 4.19-5.26). Alterations in egg coloration, melanization and granule accumulation were observed at 24 h, persisting until 144 h. The embryo digestive systems were severely damaged, including narrowing of the intestinal lumen, vesiculations and degenerated cells, causing embryonic death. CONCLUSION The toxicity caused by Bta in D. saccharalis embryos demonstrated its potential as a biological control agent and as a sustainable alternative for integrated management of D. saccharalis infestation. © 2020 Society of Chemical Industry.

Endophytic microorganisms live inside the plants without causing any damage to their hosts. In the agricultural field, these endophytes might be a strategy of biological control for phytopathogens. We aimed to isolate endophytic fungi from yellow passion fruit (Passiflora edulis) leaves, evaluating its biocontrol capacity by in vitro antagonism against phytopathogen Colletotrichum sp. CNPU378. We also carried out greenhouse experiments in bean seedlings. A high colonization frequency was obtained (89%), and the molecular identification based on DNA sequencing attested Colletotrichum as the most frequent genus and minor occurrence of Curvularia endophytes. The endophytes tested showed different types of competitive interactions in in vitro antagonism inhibition rate ranging from 28.8 to 48.8%. There were 10 promising antagonists tested for their antagonist activity of crude extracts of secondary metabolites, in which strain PE-36 (20.8%) stood out among the other strains evaluated. In the greenhouse assay, plants inoculated only with endophyte Colletotrichum sp. PE-36 was symptomless and suggest that the endophyte strengthened the growth promotion in common bean plants, especially in the root length and number of leaves when compared to control plants and other treatments. Despite many fungi of Colletotrichum genus being described as causative agents of anthracnose, in this study, the plant sampled was colonized predominantly by Colletotrichum endophytes living in asymptomatic relationship. By the way, we come across a Colletotrichum sp. endophyte able to antagonize a Colletotrichum sp. pathogen.

Neem (Azadirachta indica) is a plant native from India and very adaptable to hot climates. Commonly it has been found as an ornamental plant or used in popular medicine due to its herbal properties. The current work is a short review that aimed to discuss Neem' biotechnological application in agriculture, environment and pharmaceutical fields. The researches were performed using online databases and the papers that discussed the prospection of Neem in the agriculture field (as a fertilizer or playing plant protection against pests and disease), pharmaceutical industry (production of metabolites with biological activity) and environmental biotechnology (eco-friendly alternatives to mitigate chemical pesticide environ damage) have chosen. Based on Neem's properties is possible find it as a fertilizer, supplement for animal feed or playing important role in plant defenses against insects (bioinsecticide) and pathogenic microorganisms (antimicrobial activity) in the agriculture field or the pharmaceutical industry with metabolites which have antitumoral, antioxidant and anti-bactericidal activity, besides Neem has already been reported used in combination with nanoparticles. Overall, Neem has been mentioned as a great tool for biotechnological applications in different areas.

Larval therapy (LT) comprises the application of sterile Calliphoridae larvae for wound debridement, disinfection, and healing in humans and animals. Larval digestion plays a key role in LT, where the salivary glands and gut produce and secrete proteolytic and antimicrobial substances. The objective of this work was to bioprospect the salivary glands of Chrysomya megacephala (Fabricius, 1794) larvae, using ultrastructural, morphological, and histological observations, and the total protein electrophoretic profile. The salivary glands present a deferent duct, originating from the buccal cavity, which bifurcates into efferent ducts that insert through a slight dilatation to a pair of tubular-shaped tissues, united in the region of fat cells. Histologically, the secretion had protein characteristics. Cell cytoplasm presented numerous free ribosomes, autophagic vacuoles, spherical and elongated mitochondria, atypical Golgi complexes, and dilated rough endoplasmic reticulum. In the apical cytoplasm, secretory granules and microvilli secretions demonstrated intense protein synthesis, basal cytoplasm with trachea insertions, and numerous mitochondria. The present work described the ultrastructure and morphology of C. megacephala third instar salivary glands, confirming intense protein synthesis and the molecular weight of soluble proteins.

Made available in DSpace on 2019-10-06T16:47:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-11-30 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The relevance of Bacillus thuringiensis in pest control strategies have been increasing in recent decades. This entomopathogenic microorganism is considered safe, and its action is highly selective to target organisms. Its toxic effect is attributed to the toxins synthesized during sporulation. The nature of the produced toxins depends on the bacterial variety. Some varieties of B. thuringiensis can synthesize from one to eight distinct toxins, which have specific effects on different orders of susceptible insects. The sugarcane borer, Diatraea saccharalis, is a holometabolous lepidopteran that causes severe damage to sugar cane cultures during its larval phase. Therefore, to evaluate B. thuringiensis serovar Aizawai GC-91 efficiency and its effects on the midgut of first instar D. saccharalis larvae, we reared the insects with contaminated artificial diet for 144 h. Larvae mortality during this period revealed that 5, 10, and 20 g/L concentrations were the most efficient, resulting in 100% mortality of treated larvae, with LC50 and LC90 estimated at 0.307 and 1.330 g/L, respectively. Furthermore, histological and ultrastructural analyses revealed damage to midgut cells. Within the first 24 h of treatment, the midgut of infected insects presented peritrophic membrane degeneration, cytoplasmic vacuolization, and cellular hypertrophy; spherites and calcium granules were also observed. At 48 h of treatment, the digestive system had collapsed, with microvilli degeneration, basement membrane and muscular fiber bundles disruption, and cellular lysis. These results confirm the toxicity of the B. thuringiensis serovar Aizawai GC-91 on D. saccharalis larvae and its potential use as a biocontroller against this pest species. Laboratory of Biological Control Morphology and Insect Cytogenetics Department of Biotechnology Genetics and Cell Biology State University of Maringá - UEM Laboratory of Insects Department of Morphology Institute of Biosciences of Botucatu São Paulo State University - UNESP Microscopy Center Complex of Research Support Centers of the State University of Maringá - UEM Laboratory of Animal Genetics Department of Biotechnology Genetics and Cell Biology State University of Maringá - UEM Laboratory of Microbiology Department of Biotechnology Genetics and Cell Biology State University of Maringá - UEM Laboratory of Insects Department of Morphology Institute of Biosciences of Botucatu São Paulo State University - UNESP

Tribolium castaneum Herbst 1797 (Coleoptera: Tenebrionidae), an important pest of stored grains and byproducts, is naturally infected by Gregarina cuneata Stein 1848 (Apicomplexa: Gregarinidae). Changes in the life cycle of insects caused by the parasite development in the midgut were studied. Trophozoites, gamonts (solitary and associated), and gametocysts were present in the midgut of the insects. In young trophozoites, the apical region differentiated into an epimerite that firmly attached the parasite to the host epithelial cells. With maturation, trophozoites developed in gamonts that were associated with the initiation of sexual reproduction in the cell cycle, culminating in the formation of the spherical gametocyst. Morpho-functional analyses indicated that gregarines absorb nutrients from infected cells and can occlude the midgut as they develop. Consequently, nutritional depletion may interfere with the host's physiology, causing decreased growth, delayed development, and high mortality rates of the parasitized insects. These results suggest G. cuneata could be an important biological agent for controlling T. castaneum in integrated pest management programs.

Tribolium castaneum (Coleoptera: Tenebrionidae) is a common pest of stored grains and byproducts and is normally infected by Gregarina cuneata (Apicomplexa: Eugregarinidae). The life cycle of this parasite includes the sporozoite, trophozoite, gamont, gametocyte, and oocyst stages, which occur between the epithelium and lumen of the host's midgut. This study aims to describe the morphofunctional alterations in the midgut and determine the esterase patterns in T. castaneum when parasitized by gregarines. To achieve this purpose, midguts of adult insects were isolated, processed, and analysed using light and electron microscopy. We determined total protein content, amylase activity, and the expression and related activities of the esterases by using polyacrylamide gel electrophoresis (PAGE). The midgut of T. castaneum is formed by digestive, regenerative, and endocrine cells. The effects of parasitism on the digestive cells are severe, because the gregarines remain attached to these cells to absorb all the nutrients they need throughout their development. In these cells, the most common alterations observed include expansion and fragmentation of the rough endoplasmic reticulum, development of the smooth endoplasmic reticulum, changes in mitochondrial cristae, cytoplasmic vacuolization, formation of myelin structures, spherites, large intercellular spaces, autophagic vesicles, expansion of the basal labyrinth, and cytoplasmic protrusions. Deposits of glycogen granules were also observed. Amylase activity was reduced in parasitized insects. Regenerative cells were found in disorganized crypts and did not differentiate into new cells, thus, compromising the restoration of the damaged epithelium. Though few morphological alterations were observed in the endocrine cells, results suggest that the synthesis and/or release of hormones might be impaired. Nine esterases (EST-1 to 9) were identified in the midgut of T. castaneum and were expressed in varying levels in response to parasitism. Two additional isoforms of esterases were exclusively identified in the parasitized insects. The results of this study suggest that gregarines alter the morphology and physiology of the midgut. The changes may result in nutritional depletion and the impairment of other physiological processes, such as reproduction and development of the host. Thus, further studies are needed to uncover the possibility of utilizing gregarines as biological controllers of the insect pest population.

Grande parte dos efluentes produzidos por indústrias e domicílios são lançados no ambiente tornando-se altamente nocivos aos seres vivos e ao ambiente. Muitos destes efluentes apresentam em sua composição metais pesados. Os metais pesados podem favorecer a inativação de algumas enzimas essenciais para o metabolismo celular causando diversos danos à saúde. Em efluentes líquidos há diversas técnicas que podem ser empregadas na remoção desses contaminantes, entretanto, alguns desses métodos apresentam custo elevado, baixa remoção do metal e alta demanda de reagentes. Uma perspectiva crescente, é o uso de materiais de origem biológica para remoção dessas substâncias nocivas. Neste trabalho, buscamos realizar uma revisão de literatura (1997-2019) acerca de métodos alternativos, especialmente os biotecnológicos como biossorção e fitorremediação, para remoção de metais pesados do meio aquoso. Tais técnicas foram escolhidas por apresentarem baixo custo associado e serem ecologicamente favoráveis. Para biossorção, encontramos diferentes subprodutos de origem vegetal capazes de remover diversos íons metálicos, como a casca de laranja, coco verde, folhas de bananeira e sementes. Fungos (Pleurotus ostreatus), bactérias (Bacillus licheniformis) e microalgas (Spirulina platensis) também foram descritos como bons removedores de metais pesados. As plantas, por meio da fitorremediação, também demonstraram-se eficazes, notadamente as espécies: Lemna aequinoctialis, Helianthus annuus, Commelina benghalensis. Zea mays, Amaranthus hybridus, Pistia stratiotes. Nesta perspectiva, podemos concluir que os processos de biossorção e fitorremediação empregando produtos/subprodutos vegetais, microbianos ou animais são viáveis pelo seu baixo custo e boa disponibilidade, além de apresentarem taxas expressivas na remoção de substâncias recalcitrantes, notadamente de metais pesados.

Endófitos são fungos e bactérias que vivem no interior das plantas sem causar nenhum dano aos seus hospedeiros. Nas últimas décadas o potencial biotecnológico desses micro-organismos tem aumentado com o advento da nanotecnologia. Estudos descobriram que fungos e bactérias são capazes de produzirem nanopartículas por meio de rotas intra ou extracelulares, processo este que passou a ser descrito como micosíntese. O interesse cada vez maior para métodos mais sustentáveis e biológicos resultou no desenvolvimento de nanopartículas não-tóxicas e comparativamente mais bioativas. Ao contrário dos métodos físicos e químicos, a micosíntese é rentável e favorável ao meio ambiente. Neste trabalho buscamos realizar uma revisão de literatura acerta dos micro-organismos endofíticos e síntese biológica de nanopartículas. Os gêneros de bactérias endofíticas mais estudados quanto a micosíntese destacam-se Bacillus e Pseudomonas, enquanto para fungos Aspergillus, Penicillium, Fusarium, Colletotrichum, Altrnaria e Trichoderma. Dentre as principais aplicações biotecnológicas destas nanopartículas oriundas da síntese microbiana temos atividades antibacterianas, citotóxicas e antioxidantes. A associação entre endófitos e nanopartículas está em fase exploratória para compreender com clareza os mecanismos responsáveis por essa interação. Entretanto, sabe-se que estas nanopartículas produzidas por meio da micosíntese já são prospectadas em algumas atividades biológicas. Espera-se que todas as espécies de plantas abriguem endófitos, os quais podem ser isolados e bioprospectados para a produção de novas ou já conhecidas nanopartículas.

In Diatraea saccharalis larvae, the Malpighian tubules are found along the digestive tube, extending from the middle mesenteric region to the end of the posterior intestine, where they come in contact with the rectum to form the cryptonephridium. Scanning and transmission electron microscopy of non-parasitized and parasitized larvae by Cotesia flavipes have indicated that the tubules consist of secretory and reabsorption cells. In parasitized larvae, the occurrence of hemocytes and teratocytes around the tubules is indicative of their role in immunological defense; however, they were not observed in non-parasitized larvae. At day 9 of parasitism, the mitochondria-containing vacuoles and myelin-like figures show signs of degeneration. The results of this study have confirmed that C. flavipes manipulates the physiology and biochemistry of D. saccharalis because the Malpighian tubules of the parasitized larvae remain active until the parasitoid completes its pupal stage and is released from the host organism.

Sitophilus zeamais Motschulsky, 1855 (Coleoptera: Curculionidae) is one of the most aggressive pests of stored grains, causing significant decrease in the nutritional quality of the grains and major losses in economic trade. The ability of this pest to damage grains and other products is directly related to the morphology of the alimentary canal. Considering the importance of the foregut in the digestion, this study aimed to describe the morphology of the crop and proventriculus in S. zeamais adults. The tissues were isolated, processed, and analyzed by using light microscopy, scanning electron microscopy, and transmission electron microscopy. The crop functioned as a storage organ, and its cells showed functional characteristics related to protein synthesis. High densities of prokaryotic microorganisms and spicules were found in the lumen. The proventriculus exhibited eight chitinous teeth, which were responsible for grinding and filtering food particles. This organ formed the stomodeal valve, which controlled the unidirectional flow of food through the alimentary canal. The alimentary canal is the primary surface of contact between the external environment and an insect's internal environment, and knowledge of its morphology is required to better understand the physiology of stored-grain insect pests.

The strains MT and E9 of Metarhizium anisopliae were found to be effective in causing the morphological changes in pupae of Diatraea saccharalis of different ages. However, during the treatment with the two strains, it was observed that there was a decrease in the longevity of old pupae and adults that emerged from old and young pupae. The adults that emerged died 24 h later and were totally covered by the pathogen’s mycelium. Regarding the mortality of pupae of different ages, but in relation to the abnormalities, the pre-pupae were found to be more susceptible. The morphological alterations observed in the cuticle and in the internal tissues of the pupae were analyzed using a stereo-microscope, a light microscope and a scanning electron microscope. The adhesion stage and formation of the appressorium happened 18 h after the infection, occasioning black spots and depressions in the cuticle of young and old pupae. In pre-pupae, the sclerotization was not observed and the fungus acted faster and more effectively, thereby, achieving 100% mortality. The colonization of the pathogen was observed in young pupae between 48 and 120 h. The alterations were observed in the fat body, digestive system and musculature; moreover, several vacuoles were noticed inside the pupae. The extrusion happened after 120 h, presenting the mycelial growth upon the pupae cuticle and further sporulation upon the corpses. The morphological alterations that took place during the development of M. anisopliae suggest the possible causes for the mortality of the pupae of D. saccharalis. Key words: Biological control, entomopathogenic fungi, pupal stage, sugarcane borer.

Strongyloides stercoralis is a helminth parasite that can infect millions of people worldwide, particularly in tropical, subtropical and temperate regions with poor sanitation. Several aspects of epidemiology, biology and host-parasite interactions of S. stercoralis have been studied, and substantial knowledge has been acquired; however, very few studies on immunotherapeutic control strategies to prevent infection and disease in humans have been conducted. Therefore, this article reviews the current progress and targets toward vaccine and passive immunization approaches for Strongyloides spp.

Diatraea saccharalis (Fabricius) has cryptonephridial type Malphigian tubules (MT). This type of MT is characterized by the penetration of the distal part of the MT into the external walls of the rectum, which is usually lined with a perinephral membrane. The MT is divided into three differentiated regions: proximal, middle and distal. In this study, our objective was to compare the nuclear activities of each one of the three regions of the D. saccharalis MT by using a nuclear basophilic technique and critical electrolyte concentration with a toluidine blue stain at pH 4.0. This method allows differentiation of DNA/protein complexes in in situ and in vitro chromatin. MT chromatin structure in D. saccharalis is variable. Fifth instars have a more decondensed chromatin than fourth instars. The distal tubule region was the most decondensed region of the MT. Our data show an elevated genetic activity of the MT in the pre-metamorphosis period. The distal region of the MT has the highest observed activity, which may be associated with the re-absorption of useful components and the excretion of waste materials.

A cytopathological methodology was used to analyze infection by Bombyx mori multiple nucleopolyhedrovirus (BmMNPV), a geographic isolate of the family Baculoviridae, in the caterpillar testes of the B. mori. Japanese B. mori strain caterpillar, fifth instar, were inoculated with BmMNPV and their testes were collected and processed for light and transmission electronic microscopy. Epithelial coating cells and interfollicular septa in testes were susceptible to BmMNPV. The first evidence of infection was detected on the 6th day post-inoculation (p.i.) in the external epithelium, and on the 7th day p.i. in the internal epithelium and interfollicular septa. Cytopathological characteristics consisted of hypertrophied nuclei, the formation of virogenic stroma, and the occlusion of virions in polyhedron protein crystals in several stages of development. At the end of the infectious process, cell lysis and release of polyhedra into the extracellular medium occurred. Histopathology revealed early infection foci in the surrounding regions of tracheal insertions, thus underlining the role of the trachea as an infection-spreading organ in insects. This spreading occurs through penetration of the basal lamina, which facilitates entry of the budded virus into the testis. Additionally, an alignment of a partial sequence of the ORF 14 of the BmMNPV geographic isolate with other NPV certified the virus genera.

Diatraea saccharalis, the main pest of sugarcane, has been controlled by Cotesia flavipes. Very little is known about the effect of parasitism on the host organs, including the midgut. The Lepidoptera midgut epithelium is composed of columnar, goblet, regenerative, and endocrine cells. Spherites have been described in columnar and regenerative cells of several Lepidoptera species, and presented a lot of functional meaning. We identified spherites in the midgut epithelial cells of non-parasitized D. saccharalis larvae analyzed the effect of parasitism on spherite morphology and distribution along the length of the midgut. Midgut fragments of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. All the midgut epithelial cells showed spherites, but they were not preferentially located in a particular part of the cells. Parasitized larvae had more spherites, mainly in the columnar cells, than non-parasitized larvae. This observation was associated with an ionic imbalance within the insect host. Spherites were more abundant in the anterior midgut region than in other regions, which suggests that this region is involved in ion transport by intracellular and/or paracellular route. The morphological variability of spherites in the cells of parasitized larvae was related to the developmental stages of these structures.

Embryonic development in Diatraea saccharalis F. (Lepidoptera: Crambidae) has been studied by means of whole mounts, histochemical techniques, and light microscopy. Three embryonic stages were identified up to larval eclosion. In the first stage, 0–10 h, intense multiplication of energids occurred, and significant changes in RNA levels were detected in distinct zones before the start of blastulation. During the second stage, 10–24 h, new protein and RNA sites were detected in the egg’s periphery. This was probably because of migration of energids and formation of the blastula. Gastrulation occurs during this stage and is characterized by intense synthetic and mitotic processes. The third stage, 25–168 h, showed ortochromatic and metachromatic regions at the posterior pole and in the yolk, egg periphery, and body of the embryo. Organogenesis closes approximately on the sixth day when the embryo starts feeding on the remaining yolk. Its development is complete by the end of the seventh day on which the larva ecloses.

The anatomy and histology of the Malpighian tubules of Diatraea saccharalis larvae have been studied by stereoscope, light and scanning electron microscopy. Tubules lie between hindgut and mid-midgut; and may be subdivided into proximal, middle and distal region. Two ampullae inserted laterally before the pylorus show 2 tubules derived from each. One tubule bifurcates into 2 in the third abdominal segment; these loop backward to the hindgut to form the cryptonephridial system. Whereas larvae have 6 complete tubules, 3 on each side, histological analysis reveals 1 layer containing 2 cells, forming the lumen, microvilli at the top of cells, tracheae and tracheoles, and crystals in the tubule lumen, mainly in the middle and distal regions.

Aedes mosquitoes are known to be infected by arboviruses causing disease such as dengue, zika fever, and chikunguya fever, and subsequently transmit them, to humans through the bite of infected females. Chemical control is a measure adopted as part of sustainable management and integrated vector control for public health. There are four principal classes of insecticides used for controlling mosquitoes, all being neurotoxic: organochlorides, organophosphates, carbamates, and pyrethroids. The objective of this work was to review reports on the environmental effects of the insecticides most commonly used for controlling Ae. aegypti . This bibliographic study was conducted using articles and books available in the literature with no time restriction. The databases accessed were: Google Scholar, Pub med, SciELO, and ScienceDirect. These insecticides exhibit toxicity to the environment, and may accumulate in food and water and in the body of vertebrates. Resistance to different insecticides is a problem when the mode of control is chemical, because insects survive insecticide application and higher doses are necessary for controlling the vectors. Considering these results, the ideal method would be the newly proposed means of mosquito control using technology related to modern biotechnology .

Sitophilus zeamais is one of the most aggressive pests of stored grains, causing a significant decrease in the nutritional quality of the grains and major losses in economic trade. The foraging capacity of this pest is assigned to its highly efficient digestive system. Investigations on the morphofunctional features of the midgut, which is the most active region of the alimentary canal, are fundamental to understand the feeding habits of this species. In this study, the midgut of adult insects was isolated, processed, and analyzed on light microscopy, scanning and transmission electron microscopy, protein and enzymatic activities determination, including analyses of the starch hydrolysis products. In S. zeamais, the midgut was differentiated into anterior midgut and posterior midgut, and consisted of digestive, regenerative and endocrine cells. The anterior midgut showed high density of regenerative crypts. Cells containing organelles associated with protein synthesis and presence of amylases and lipases indicated that majority of the digestion process occurred in the anterior midgut. The posterior midgut exhibited numerous gastric caeca and peritrophic membrane. Cells with poorly differentiated cytoplasmic into organelles, elongated microvilli, and low enzymatic activities indicated that the posterior midgut was mainly involved in absorption.

The effects of a commercial formulation of oil from neem, Azadirachta indica A. Juss (Meliaceae), on the eggs of sugarcane borer Diatraea saccharalis F.,1794 (Lepidoptera: Crambidae), at different stages of its embryonicdevelopment were investigated. To evaluate the ovicidal activity of the oil, eggs were sprayed with the product at concentrations of 0.1%, 0.3%, 0.5%, 1.0% and 2.0% at 1, 2, 3, 4, 5 and 6 days after day were laid. Controls were sprayed with distilled water. The neem oil was toxic to eggs, mainly when applied to 2-, 3-, 4- and 5-daysold eggs. The percentage of viability reduction was 31-99%. Higher levels of caterpillar eclosion were obtained in eggs treated with neem oil at older ages. However, the resulting caterpillars had fatal morphological anomalies, except those exposed to 0.1% neem oil. Neem oil is highly toxic when sprayed on D. saccharalis eggs, as demonstrated by severely reduced hatching, increased mortality of hatched caterpillars, impaired embryonic development, and residual activity in the production of new individuals. Therefore, neem oil may be a promising agent against the sugarcane borer, during the stage in which penetrates the stalk, causing heavy damage to the sugar-cane crop.

The continuous growth of world population and the increased use of pesticides became obvious environmental problems, leading to agricultural production to develop alternative practices that promote environmental sustainability. The use of biological control techniques to reduce agricultural pests have been increasing, enabling the substitution of conventional insecticides by diversified natural enemies such as insects, viruses, fungi, bacteria, plant extracts, and others. A quantitative-descriptive analysis was carried using a questionnaire as mean to personal interview of organic growers in the city of Maringa, state of Paraná, Brazil was held. The aim was quantify about the knowledge of biological controllers by this growers, the use, and economic viability of these, aiming to promote the expansion of the practice and development of new controllers to replace or reduce the use of chemicals. The results show that all growers have the knowledge and have used alternative methods of biocontrol, some of them consider the practice of great environmental importance, but is still required further studies of the regional entomofauna to better effectiveness of the biocontrol.

Diatraea saccharalis, the main pest of sugarcane, has been controlled by Cotesia flavipes. Very little is known about the effect of parasitism on the host organs, including the midgut. The Lepidoptera midgut epithelium is composed of columnar, goblet, regenerative, and endocrine cells. Spherites have been described in columnar and regenerative cells of several Lepidoptera species, and presented a lot of functional meaning. We identified spherites in the midgut epithelial cells of non-parasitized D. saccharalis larvae analyzed the effect of parasitism on spherite morphology and distribution along the length of the midgut. Midgut fragments of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. All the midgut epithelial cells showed spherites, but they were not preferentially located in a particular part of the cells. Parasitized larvae had more spherites, mainly in the columnar cells, than non-parasitized larvae. This observation was associated with an ionic imbalance within the insect host. Spherites were more abundant in the anterior midgut region than in other regions, which suggests that this region is involved in ion transport by intracellular and/or paracellular route. The morphological variability of spherites in the cells of parasitized larvae was related to the developmental stages of these structures.

The migration rate of esterases and their substrate specificity for 4-methylumbelliferyl esters (acetate, propionate, and butyrate) and alpha- and beta-naphthyl esters were analyzed in Diatraea saccharalis by starch gel electrophoresis. Substrate preference of esterases was observed with Est-2 and Est-8 isozymes showing substrate specificity for 4-methylumbelliferyl esters and Est-4 isozyme showing specificity for 4-methylumbelliferyl butyrate and alpha-naphthyl butyrate. Allele variation was detected at the Est-3 locus. Two alleles, Est-3F and Est-3S, were identified in pupae with fluorogenic and ester-naphthyl substrates. Chi-square analysis showed no differences between the observed genotypic frequencies and those expected on the basis of Hardy-Weinberg frequencies for the Est-3 locus (chi² = 2.4; p < 0.01). The negative value for the Wright's fixation index (F = -0.2096) calculated for the D. saccharalis population maintained under laboratory conditions indicates an excess of heterozygotes, however, the observed Hardy-Weinberg equilibrium indicates that in the laboratory the population of D. saccharalis behaved as if the moth were randomly mating in nature. The high level of heterozygosity at the Est-3 locus indicates also that this esterase may be a good genetic marker for studies of natural D. saccharalis populations.

A diversidade do meio ambiente esta diretamente relacionada com equilibrio ambiental, o qual e diretamente afetado pela liberacao de residuos industriais e urbanos. Algumas especies de microrganismos podem atuar como decompositores, em processos de biorremediacao, auxiliando na recuperacao de areas contaminadas. O municipio de Maringa, localizado no estado do Parana – Brasil, vem apresentando rapido crescimento populacional e se destacando principalmente nos setores de construcao civil, textil, entre outros. Como consequencia, ha o aumento de residuos gerados e, desta forma, a necessidade de implantacao de novas tecnologias de tratamentos de residuos a fim de reduzi-los, de maneira a diminuir o impacto socio-economico-ambiental. A cidade conta com o Plano de Gerenciamento de Residuos (PGR), que visa o controle dos residuos produzidos e a sua destinacao correta. Com o intuito de melhorar e buscar alternativas para o tratamento dos residuos gerados, o objetivo deste trabalho foi relacionar os dejetos industriais e domesticos produzidos no municipio de Maringa com possiveis formas de tratamento utilizando, de maneira sustentavel, o potencial biorremediador de microrganismos.