Analysis of mycobiota linked to insects isolated from pet food collected in pet shops / Análise da micobiota associada à entomofauna em rações a granel para animais domésticos

Insects invade stored foods such as corn, nuts and dried fruits that are ingredients of pet food. Toxigenic fungi (Aspergillus, Penicillium and Fusarium) are also contaminants of these substrates. The insects are mechanical vectors and the infestation increases the humidity of dried food, thus promoting the fungal growth and the production of mycotoxins. The objective of this study was to analyze the mycobiota associated with insect infestation found in pet foods. Twenty samples were plated on potato agar incubated at 25º C during 7 days, for the isolation of fungi from pet food and from insects isolated from each ingredient. The insects were identified by using a dichotomous key. All samples showed fungal presence, and Sitophilus zeamais was the predominant insect with the highest frequency in the collected samples, indicating it as a vector of many fungal genera. Pet foodscomposed by a mixture of seeds and grains are subject to microbial contamination, and the insects are important mechanical vectors of spoilage and toxigenic fungi, endangering animal health.

Insetos podem infestar sementes e grãos que integram as rações destinadas à alimentação de pequenos roedores e aves domésticas como amendoim, girassol, trigo, milho, aveia e outros. Fungos toxigênicos (Aspergillus, Penicillium e Fusarium) também são contaminantes destes substratos e podem ser carreados por insetos. A deterioração da massa de sementes e grãos pela atividade dos insetos associado à presença de umidade, pode ser um facilitador para o crescimento fúngico e produção de micotoxinas. O objetivo do estudo foi o de analisar a micobiota associada às sementes, grãos e insetos em ração mista para animais domésticos. Vinte amostras foram plaqueadas em ágar-batata-dextrose incubadas a 25º C durante 7 dias para o isolamento fúngico de ração e em insetos isolados de cada componente da ração. Todas as amostras apresentaram presença de fungos e Sitophilus zeamais, inseto predominante e com maior frequencia nas amostras coletadas, demonstrando ser um vetor de vários gêneros fúngicos. Rações animais compostas de misturas de sementes e grãos estão sujeitas à contaminação microbiológica, sendo os insetos importantes vetores mecânicos de fungos deteriorantes e toxigênicos, comprometendo a saúde animal.

Analysis of mycobiota linked to insects isolated from pet food collected in pet shops / Análise da micobiota associada à entomofauna em rações a granel para animais domésticos

Insects invade stored foods such as corn, nuts and dried fruits that are ingredients of pet food. Toxigenic fungi (Aspergillus, Penicillium and Fusarium) are also contaminants of these substrates. The insects are mechanical vectors and the infestation increases the humidity of dried food, thus promoting the fungal growth and the production of mycotoxins. The objective of this study was to analyze the mycobiota associated with insect infestation found in pet foods. Twenty samples were plated on potato agar incubated at 25º C during 7 days, for the isolation of fungi from pet food and from insects isolated from each ingredient. The insects were identified by using a dichotomous key. All samples showed fungal presence, and Sitophilus zeamais was the predominant insect with the highest frequency in the collected samples, indicating it as a vector of many fungal genera. Pet foodscomposed by a mixture of seeds and grains are subject to microbial contamination, and the insects are important mechanical vectors of spoilage and toxigenic fungi, endangering animal health.(AU)

Insetos podem infestar sementes e grãos que integram as rações destinadas à alimentação de pequenos roedores e aves domésticas como amendoim, girassol, trigo, milho, aveia e outros. Fungos toxigênicos (Aspergillus, Penicillium e Fusarium) também são contaminantes destes substratos e podem ser carreados por insetos. A deterioração da massa de sementes e grãos pela atividade dos insetos associado à presença de umidade, pode ser um facilitador para o crescimento fúngico e produção de micotoxinas. O objetivo do estudo foi o de analisar a micobiota associada às sementes, grãos e insetos em ração mista para animais domésticos. Vinte amostras foram plaqueadas em ágar-batata-dextrose incubadas a 25º C durante 7 dias para o isolamento fúngico de ração e em insetos isolados de cada componente da ração. Todas as amostras apresentaram presença de fungos e Sitophilus zeamais, inseto predominante e com maior frequencia nas amostras coletadas, demonstrando ser um vetor de vários gêneros fúngicos. Rações animais compostas de misturas de sementes e grãos estão sujeitas à contaminação microbiológica, sendo os insetos importantes vetores mecânicos de fungos deteriorantes e toxigênicos, comprometendo a saúde animal.(AU)

Anti-Trypanosoma cruzi effects of cyclosporin A derivatives: possible role of a P-glycoprotein and parasite cyclophilins.

Cyclophilins are target molecules for cyclosporin A (CsA), an immunosuppressive antimicrobial drug. We have previously reported the in vitro anti-Trypanosoma cruzi activity of H-7-94 and F-7-62 non-immunosuppressive CsA analogues. In this work, we continue the study of the parasiticidal effect of H-7-94 and F-7-62 CsA analogues in vitro and in vivo and we analyse 3 new CsA derivatives: MeIle-4-CsA (NIM 811), MeVal-4-CsA (MeVal-4) and D-MeAla-3-EtVal-4-CsA, (EtVal-4). The most efficient anti-T. cruzi effect was observed with H-7-94, F-7-62 and MeVal-4 CsA analogues evidenced as inhibition of epimastigote proliferation, trypomastigote penetration, intracellular amastigote development and in vivo T. cruzi infection. This trypanocidal activity could be due to inhibition of the peptidyl prolyl cis-trans isomerase activity on the T. cruzi recombinant cyclophilins tested. Furthermore, CsA and F-7-62 derivative inhibited the efflux of rhodamine 123 from T. cruzi epimastigotes, suggesting an interference with a P-glycoprotein activity. Moreover, H-7-94 and F-7-62 CsA analogues were not toxic as shown by cell viability and by aminopyrine-N-demethylase activity on mammalian cells. Our results show that H-7-94, F-7-62 and MeVal-4 CsA analogues expressed the highest inhibiting effects on T. cruzi, being promissory parasiticidal drugs worthy of further studies.

Analysis of the Trypanosoma cruzi cyclophilin gene family and identification of Cyclosporin A binding proteins.

The Trypanosoma cruzi cyclophilin gene family comprises 15 paralogues whose nominal masses vary from 19 to 110 kDa, namely TcCyP19, TcCyP20, TcCyP21, TcCyP22, TcCyP24, TcCyP25, TcCyP26, TcCyP28, TcCyP29, TcCyP30, TcCyP34, TcCyP35, TcCyP40, TcCyP42 and TcCyP110. Under the conditions used, only some of the T. cruzi cyclophilin paralogue products could be isolated by affinity chromatography. The 15 paralogues were aligned with 495 cyclophilins from diverse organisms. Analyses of clusters formed by the T. cruzi cyclophilins with others encoded in various genomes revealed that 8 of them (TcCyP19, TcCyP21, TcCyP22, TcCyP24, TcCyP35, TcCyP40, TcCyP42 and TcCyP110) have orthologues in many different genomes whereas the other 7 display less-defined patterns of their sequence attributes and their classification to a specific group of cyclophilin's orthologues remains uncertain. Seven epimastigote cDNA clones encoding cyclophilin isoforms were further studied. These genes were found dispersed throughout the genome of the parasite. Amastigote and trypomastigote mRNAs encoding these 7 genes were also detected. We isolated 4 cyclosporin A-binding proteins in T. cruzi epimastigote extracts, which were identified by mass spectrometry as TcCyP19, TcCyP22, TcCyP28 and TcCyP40. Cyclosporin A-binding to these cyclophilins might be of importance to the mechanism of action of Cyclosporin A and its non-immunosuppressive analogues, whose trypanocidal effects were previously reported, and therefore, of potential interest in the chemotherapy of Chagas' disease.

Avaliação de produtos naturais para o controle do ácaro rajado tetranychus urticae (koch, 1836) (acari: tetranychidae) em casa de vegetação

ABSTRACT Tetranychus urticae is considered one of the most important mite species, attacking several agricultural crops. The aim of this work was to verify, in the greenhouse, the acaricide activity of some plant extracts on a population of T. urticae, infesting bean plants. Each plot consisted of 5 bean plants with 7 to 10 leaves previously infested with 5 T. urticae females each. The plant species used were Allamanda cathartica, Dieffenbachia brasiliensis, Pennisetum purpureum, Annona squamosa, Ruta graveolens, Sonchus oleraceus, Spondias purpurea, Codiaeum variegatum, Impatiens walleriana, Stryphnodendron barbatiman, Solanum melongena, Agave angustifolia and Allium cepa. The vegetable material collected was dried in an oven at 40º C and after that triturated, and the resulting dust was submitted to extraction with distilled water for 14 hours and then filtered. The filtered residues were utilized for tests after a period of 30 to 45 days of infestation, when they were applied on infested plants in doses of 100 mL of extract/L of water utilizing a knapsack sprayer of 5 L. Previously and 7 days after treatment an evaluation was made of the number of live mites (active stages) on 1cm2 of 10 leaves in each plot. The aqueous extracts of D. brasiliensis, R. graveolens, A. cepa, A. angustifolia and A. squamosa promoted a significant reduction of the T. urticae population, showing 86.87%, 83.95%, 80.97%, 76.30 and 75.40% efficiency, respectively.

RESUMO O ácaro rajado Tetranychus urticae é considerado um dos ácaros de maior importância por atacar várias culturas. O objetivo desse trabalho foi verificar, em casa de vegetação, a ação acaricida de alguns extratos vegetais sobre população de T. urticae, infestando plantas de feijão. Cada parcela constou de 5 plantas de feijão com 07-10 folhas previamente infestadas com 5 fêmeas de T. urticae cada. As espécies vegetais utilizadas foram Allamanda cathartica, Dieffenbachia brasiliensis, Pennisetum purpureum, Annona squamosa, Ruta graveolens, Sonchus oleraceus, Spondias purpurea, Codiaeum variegatum, Impatiens walleriana, Stryphnodendron barbatiman, Solanum melongena, Agave angustifolia e Allium cepa. O material vegetal coletado foi seco em estufa a 40º C e posteriormente moído e o pó resultante submetido à extração com água destilada por 14h e em seguida, filtrado. Os resíduos filtrados foram utilizados nos testes após um período de 30-45 dias da infestação, sendo aplicados sobre as plantas infestadas, na dose de 100 mL de extrato/L de água, empregando-se um pulverizador costal de 5 L. Previamente e após 7 dias do tratamento, foi avaliado o número de ácaros vivos (formas móveis), em uma área de 1 cm2 em 10 folhas de cada parcela. Os extratos aquosos de D. brasiliensis, R. graveolens, A. cepa, A. angustifolia e A. squamosa promoveram redução significativa da população de T. urticae, apresentando 86,87%, 83,95%, 80,97%, 76,30 e 75,40% de eficiência, respectivamente.

Controle químico do pulgão takecallis taiwanus (takahashi) (homoptera: aphididae) em bambu ornamental (bambusa gracilis horti) / Chemical control of the aphid takecallis taiwanus (takahashi) (homoptera: aphididae) in ornamental bamboo (bambusa gracilis horti

RESUMO As plantas ornamentais compreendem centenas de espécies utilizadas em projetos paisagísticos sendo recentemente constatada a presença do pulgão Takecallis taiwanus infestando bambu Bambusa gracilis em cerca-viva. O presente trabalho teve por objetivo avaliar a eficiência dos inseticidas clorpirifós, malation e fenitrotion para o controle deste pulgão. Os inseticidas clorpirifós a 10 mL/ 10 L água, malation a 30 mL/10 L água e fenitrotion a 200 mL/10 L água, foram eficientes para o controle de T. taiwanus em B. gracilis até 21 dias após a aplicação.

ABSTRACT There are hundreds of species of ornamental plants used in landscape projects and, recently, the presence of the aphid Takecallis taiwanus was identified infesting bamboo in hedges. This work was carried out to evaluate the efficiency of chlorpyrifos, malathion and fenitrothion for the control. The treatments were: chlorpyrifos at 10 mL/10 L water, malathion at 30 mL/10 L water and fenitrothion at 200 mL/10 L water. All tested formulations were efficient for the control of aphid T. taiwanus on B. gracilis until 21 days after application.

Avaliação acaricida de produtos naturais para o controle de ácaro vermelho do cafeeiro oligonychus ilicis (mcgregor) (acari: tetranychidae) / Evaluation of plant extracts for the control of coffee red mite oligonychus ilicis (mcgregor) (acari: tetranychidae)

RESUMO O ácaro Oligonychus ilicis apresenta ampla distribuição no Estado de São Paulo, com maiores incidências nos períodos mais secos do ano. Estes ácaros possuem o hábito de estarem presentes na página superior das folhas do cafeeiro. Foram avaliados no controle deste ácaro, extratos aquosos, etanólicos e hexânicos de Lavandula angustifolia Mill., Dahlia pinnata Cav., Solanum paniculatum L., Agave angustifolia Haw, Ocimum basilicum L., Solanum melongena L., Rhododendron simsii Planch., Ficus elastica Roxb, Codiaeum variegatum (L.) Bl., Spondias purpurea L., Sonchus oleraceus L., Ruta graveolens L., Annona squamosa L., Pennisetum purpureum Schumach., Dieffenbachia brasiliensis (Veiech) e Allamanda cathartica L. Discos foliares de café var. Mundo Novo foram mergulhados na solução de extrato por 5seg e as fêmeas adultas em número de 20 por parcela confinadas na superfície do disco e mantidas em câmara incubadora a 25 ± 2º C e 70 ± 10% UR. Avaliação de mortalidade foi realizada com 48h. Os melhores resultados em eficiência foram obtidos com os extratos etanólico e hexânico de Dieffenbachia brasiliensis (56 e 52%), D. pinnata (54%), Allamanda cathartica (44%) e S. paniculatum (46 e 48%).

ABSTRACT The coffee red mite Oligonychus ilicis, is widespread in the state of São Paulo, with higher incidence on coffee plantations during the dry seasons. These mites live almost exclusively on the top side of the host leaves. Evaluations of aqueous, ethanolic and hexanic extracts were made for Lavandula angustifolia Mill., Dahlia pinnata Cav., Solanum paniculatum L., Agave angustifolia Haw, Ocimum basilicum L., Solanum melongena L., Rhododendron simsii Planch., Ficus elastica Roxb, Codiaeum variegatum (L.) Bl., Spondias purpurea L., Sonchus oleraceus L., Ruta graveolens L., Annona squamosa L., Pennisetum purpureum Schumach., Dieffenbachia brasiliensis (Veiech) and Allamanda cathartica L. Foliar discs of coffee var. Novo Mundo were immersed in the extract solution for 5 seconds and adult females (20 per replication) were confined on the surface of the disc and kept in an incubator chamber at 25 ± 2º C and 70 ± 10% RU. Survey of the mortality was taken at 48h. The best results of efficiency were obtained with the ethanolic and hexanic extracts of: Dieffenbachia brasiliensis (56 and 52%), D. pinnata (54%), Allamanda cathartica (44%) and S. paniculatum (46 and 48%).

Proteinase and gelatinolytic properties of a Triatoma infestans extract.

An extract of Triatoma infestans has previously been demonstrated to produce specific IgG and IgE both in animals and in atopic humans with rhinitis/asthma as well as hypersensitivity pneumonitis in guinea pigs aerosolized with T. infestans. We attempted to determine whether the antigen or antigens responsible belonged to the protease group, as occurs with other allergens such as house dust mites and cockroaches. To do this, T. infestans was studied by SDS-PAGE, Western blots and gelatinolysis with and without the use of specific protease inhibitors such as E-64, TLCK, TPCK, PMSF, leupeptin, o-phenanthrolene and pepstatin-A. These assays revealed serine-like proteolytic and gelatinolytic activities. The presence of 10 to 12 bands of between 14 and 100 kDa was detected. The proteolytic activity pattern of T. infestans was greatest at pH 8.5 and gelatinolytic activity was highly sensitive to PMSF, suggesting that this enzyme could be characterized as a serine protease. Western blots revealed that two bands of 17 and 58 kDA reacted with the sera of atopic humans with respiratory diseases and anti-IgE. However, whether these bands correlated with allergenicity is unclear since the presence of several proteins in each of these bands does not rule out the possibility that this correlation could exist, especially because cross-reactions with antigens from the cockroach Periplaneta americana and its specific antiserum in animals and atopic humans have been demonstrated. The role of proteases in the etiopathogenesis of perennial rhinitis and bronchial asthma in inhabitants of the area of Argentina infested by T. infestans requires further investigation.

Proteinase and gelatinolytic properties of a bat feces extract.

It was previously demonstrated that a bat feces extract (BAT) was able to produce a specific IgG in animals, a specific IgE in respiratory atopic humans and a hypersensitivity pneumonitis in guinea pigs. As numerous allergens (such as house-dust mite, cockroaches and pollens) revealed a enzymatic activity measured by different assays we decided to study the proteinase and the gelatinolytic activities of the BAT. Several protease inhibitors such as E-64, TLCK, TPCK, PMSF, leupeptin, o-phenantroline and pepstatin-A were applied to establish the chemical properties of the enzymatic activity. These assays revealed a serine-trypsin-like proteolytic and gelatinolytic activities specially at pH 8,5. On the other hand, two bands of 21 and 40 kDa reacted with the human atopic sera suggesting a possible correlation between allergenicity and proteinase activity. Their role in the etiology of perennial rhinitis and asthma requires further investigations.

[Hypersensitivity pneumonitis induced in guinea pigs by antigens to bat feces]. / Neumonitis por hipersensibilidad inducida en cobayos por los antígenos de las heces del murciélago.

A bat feces extract was able to induce the synthesis of specific IgG in rabbits and IgE in atopics. These findings were corroborated by skin tests, RAST and RAST-inhibition. Nowadays, data concerning the experimental induction of hypersensitivity pneumonitis using a glycoprotein of bat feces are presented. This antigen was aerosolized for inhalation by adult guinea pigs for 12 weeks to detect specific serum IgM, IgG and IgE antibodies as well as sensitized T-CD4 cells. Histopathological studies of the lungs showed interstitial infiltrates of macrophages and lymphocytes, cellular bronchiolitis and single non-necrotizing granulomas from the seventh to the ninth weeks. From the tenth week to the end of the experiment the lesions of the lungs progressively worsened. The results from this animal model suggest that the chronic contact with the bat feces'antigen can induce other inflammatory lung reactions than those IgE dependant.