Actividad insecticida de aceites esenciales sobre Blatella germanica (Linneaus, 1767) (Dictyoptera: Blattellidae) / nsecticidal activity of essential oils against Blatella germanica (Linneaus, 1767) (Dictyoptera: Blattellidae)

Blattella germanica (Linneaus, 1767) es una especie de cucaracha considerada plaga de la salud pública por estar asociada a gran número de microorganismos causantes de enfermedades al hombre. Para su control se utilizan diferentes tipos de formulaciones a base de insecticidas sintéticos a los cuales en su gran mayoría es resistente. En este contexto existe un interés creciente por los insecticidas botánicos. En el siguiente trabajo se evaluaron los aceites de Citrus aurantium (L.,1753), Ocimum basilicum (L.,1753), Piper aduncum subsp ossanum (C.DC. Saralegui) y Eucalyptus globulus (Labill, 1800) mediante aplicación tópica de un microlitro en el primer esternito abdominal de los individuos. Los cuatro aceites mostraron actividad insecticida sobre adultos de B. germanica con CL50 que oscilaron entre 58 µg/µL para O. basilicum y 250 µg/µL para P. aduncum(AU)´

Blattella germanica (Linneaus, 1767) is a cockroach species considered a public health pest, since it is associated with a great number of disease-causing microorganisms in humans. For its control, different types of synthetic-based insecticidal formulations are used, to which it is mostly resistant. In this context, there is a growing interest in botanical insecticides. In this research, oils from Citrus aurantium (L., 1753), Ocimum basilicum (L., 1753), Piper aduncum subsp. ossanum (C.DC. Saralegui), and Eucalyptus globulus (Labill, 1800) were evaluated by topical application of 1 µL to the first abdominal sternum of the individuals. The four essential oils evaluated showed insecticidal activity against adult B. germanica with LC50 ranging from 58µg/µL for O. basilicum to 250µg/µL for P. aduncum(AU)´

Discovery of cockroach specimens of the genus Squamoptera (Ectobiidae: Pseudophyllodromiinae) from Okinawa, Japan, showing wing polymorphism

@# Cockroach specimens of the genus, Squamoptera were collected from the Iriomote island of Okinawa prefecture, Japan. The morphological features of the specimens were characterized as having a white band on the dorsal surface of its thorax, its tegmen reduced into a tiny scale-like structure and the hindwing was absent. Ocelli was also absent and the small compound eyes not extending to apex of the head nor to the frontal face but extend further lower than the base of the antennae. When the specimens were reared in the laboratory, besides the short wing form, the long wing form began to appear in the rearing colony. In our reproductive biological study, we observed that hatching of the ootheca from the short wing female takes about 30 days, with an average of 6.6 nymphs being hatched from one ootheca. The male to female ratio of the offspring was 36:30. However, the frequency appearance of the offspring from the ootheca of the short wing female was 98.5% short wing and 1.5% long wing form. Our specimens occasionally show body polymorphism in the form of individuals having long wings instead of the usual short one. The long wing form does not show the white band on the dorsal surface of its thorax.

A new species of cockroach, Periplaneta gajajimana sp. nov., collected in Gajajima, Kagoshima Prefecture, Japan

@#We described a new species of cockroach, Periplaneta gajajimana sp. nov., which was collected in Gajajima, Kagoshima-gun Toshimamura, Kagoshima Prefecture, Japan, on November 2012. The new species is characterized by its reddish brown to blackish brown body, smooth surface pronotum, well developed compound eyes, dark brown head apex, dark reddish brown front face and small white ocelli connected to the antennal sockets. In male, the tegmen tip reach the abdomen end or are slightly shorter, while in the female, it does not reach the abdominal end and exposes the abdomen beyond the 7th abdominal plate. We confirmed the validity of this new species by breeding the specimens in our laboratory to demonstrate that the features of the progeny were maintained for several generations. For comparison and easy identification of this new species, the key to species identification of the genus Periplaneta that had been reported in Japan to date are also presented.

Do dietary stresses affect the immune system of Periplaneta americana (Blattaria: Blattidae)? / Estresses alimentares afetam o sistema immune de Periplaneta americana (Blattaria: Blattidae)?

Abstract Stresses can be caused by multiple biotic and abiotic factors and their effects can affect both the biology and the immune system of insects. American cockroach - Periplaneta americana (Linnaeus, 1758) (Blattaria: Blattidae) -besides being an excellent model species, has great medical importance because it can act as a mechanical vector of several pathogens. This study aimed to evaluate the influence of starvation, dehydration and both stresses on weight, and total and differential haemocyte count in P. americana adults. Each specimen was isolated in glass flasks containing or not food and/or water. They were weighed periodically. Another group received water for 24 h after the end of stress period. In the immunologic bioassay, we counted their haemocytes after the final weighing. All stresses reduced the insect weight, especially when the stresses were combined. Females of the control group gained weight and males had it unaltered. Different stress conditions and time did not influence on total haemocyte count. Insects without food and water had the proportion of prohaemocytes increased and plasmatocytes decreased. This study can serve as a basis of further studies of bioecology, behaviour and the ability of resisting insecticides, besides serving as a model to studies in other insect species.

Resumo Os estresses podem ser causados por múltiplos fatores bióticos e abióticos e seus efeitos podem afetar tanto a biologia como o sistema imune dos insetos. A barata-americana - Periplaneta americana (Linnaeus, 1758) (Blattaria: Blattidae) - além de ser uma excelente espécie modelo, tem grande importância médica, pois pode atuar como vetor mecânico de diversos patógenos. O objetivo desse estudo foi avaliar a influência da inanição, desidratação e ambos os estresses sobre o peso e o número total e diferencial de hemócitos em adultos de P. americana. Cada espécime foi isolado em frascos de vidro contendo ou não alimento e/ou água. Eles foram pesados periodicamente. Outro grupo recebeu água por 24 h após o término do período de estresse. Nos ensaios imunológicos, foram contados os seus hemócitos após a última pesagem. Todos os estresses reduziram o peso dos insetos, especialmente quando os estresses foram combinados. As fêmeas do grupo controle ganharam peso e os machos tiveram seu peso inalterado. As diferentes condições de estresse e tempo não influenciaram no número total de hemócitos. Os insetos sem alimento e água tiveram a proporção de pró-hemócitos aumentada e a de plasmatócitos reduzida. Esse estudo pode servir como base para estudos posteriores de bioecologia, comportamento e da habilidade de resistir aos inseticidas químicos, além de servir como modelo para estudos em outras espécies de insetos.

Surveillance of cockroach population and evaluation of control effects in households from 2017 to 2019 in Jiading District of Shanghai / 上海预防医学

Objective To learn the population and infestation rates of cockroaches from 2017 to 2019 in Jiading District of Shanghai, to evaluate the effect of cockroach termination in household, and to provide information for cockroach control. Methods Cockroaches were controlled by dinotefuran baits and clean-up in households.Sticky trap and visual method were employed for density monitoring in farmers markets, supermarkets, hotels, restaurants, hospitals, and residential areas.Visual method was used in households before and after using the insecticide. Results Sticky trap result showed the room infestation rate was 3.24%, mean adhesion rate was 3.29%, the density was 0.06 per board, and the density peak appeared in May.Rate of invasion and density decreased year by year.Blattella germanica was the dominant species, counting for 71.88%.The density, and rate of infestation, as determined by sticky trap method, were the highest in the farmers markets, followed by hospitals and residential areas.Determined by visual method, room infestation rate was 1.16%, and the infestation rate was 4.44%.The peak appeared in January.Infestation rate of the farmers markets was the highest, followed by hospitals and residential areas.By visual method, the room infestation rate was 59.01%, and 48.45% for nymphs.The room infestation and ootheca rates were 54.04% and 17.39%.The rate decreased more than 80% in 30 days after use of the insecticide. Conclusion Infestation rate of cockroach remains at low level in Jiading District.The effect of bait combined with environmental cleaning is remarkable.Future work should strengthen monitoring and control in farmers markets, hospitals and residential areas.

Laboratory study on chain-killing efficacy of four gel baits / 上海预防医学

Objective To study the first-time killing efficacy and the chain-killing efficacy of four gel baits against Blattella germanica: 1% chlorpyrifos, 0.05% fipronil, 2.15% imidacloprid, and 0.5% dinotefuran and provide a basis for drug selection in controlling Blattella germanica. Methods Laboratory killing efficacy test was conducted according to the national standard GB/T 13917.7-2009 and the chain-killing efficacy test was conducted for three rounds.The first round of chain efficacy test was conducted by feeding the cockroaches killed in the laboratory efficacy test, and each next round by feeding the cockroaches killed in the last round.Median lethal time (LT50), 95% confidence limit, and toxicological regression equation of each test were calculated by software DPS V9.01. Results The LT50 of the efficacy test with 1% chlorpyrifos gel bait was 0.745 5 (0.603 4-0.890 3) d.The LT50 of the first, second and third chain experiments increased by 3.30, 2.18 and 2.76 times, respectively.The LT50 of the efficacy test with 0.05% fipronil gel bait was 0.846 5(0.464 7-1.228 0)d, and increased by 5.42, 2.09 and 1.48 times, respectively, in the first, second and third chain experiments.The LT50 of the efficacy test with 2.15% imidacloprid gel bait was 3.192 1(2.865 0-3.506 0)d, and increased by 1.13, 1.65 and 1.15 times, respectively in the first, second and third chain experiments.The LT50 of the efficacy test with 0.5% dinotefuran gel bait was 0.997 1(0.805 8-1.191 6) d, and increased by 3.85, 1.37 and 1.78 times, respectively in the first, second and third chain experiments. Conclusion In the laboratory killing efficacy test, 1% chlorpyrifos, 0.05% fipronil, and 0.5% dinotefuran gel baits are better than 2.15% imidacloprid gel bait.In the chain-killing efficacy test, 2.15% imidacloprid and 0.5% dinotefuran gel baits are better than 1% chlorpyrifos and 0.05% fipronil gel baits.Based on our results, we recommend the use of 0.5% dinotefuran gel bait for comprehensive and sustained killing effect.

German Cockroach Extract Induces Matrix Metalloproteinase-1 Expression, Leading to Tight Junction Disruption in Human Airway Epithelial Cells

PURPOSE: Cockroach exposure is a pivotal cause of asthma. Tight junctions are intercellular structures required for maintenance of the barrier function of the airway epithelium, which is impaired in this disease. Matrix metalloproteinases (MMPs) digest extracellular matrix components and are involved in asthma pathogenesis: MMP1 is a collagenase with a direct influence on airway obstruction in asthmatics. This study aimed to investigate the mechanism by which German cockroach extract (GCE) induces MMP1 expression and whether MMP1 release alters cellular tight junctions in human airway epithelial cells (NCI-H292). MATERIALS AND METHODS: mRNA and protein levels were determined using real-time PCR and ELISA. Tight junction proteins were detected using immunofluorescence staining. Epithelial barrier function was measured by transepithelial electrical resistance (TEER). The binding of a transcription factor to DNA molecules was determined by electrophoretic mobility shift assay, while the levels of tight junction proteins and phosphorylation were determined using Western blotting. RESULTS: GCE was shown to increase MMP1 expression, TEER, and tight junction degradation. Both an inhibitor and small interfering RNA (siRNA) of MMP1 significantly decreased GCE-induced tight junction disruption. Furthermore, transient transfection with ETS1 and SP1 siRNA, and anti-TLR2 antibody pretreatment prevented MMP1 expression and tight junction degradation. An extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) inhibitor also blocked MMP1 release, ETS1/SP1 DNA binding, and tight junction alteration. CONCLUSION: GCE treatment increases MMP1 expression, leading to tight junction disruption, which is transcriptionally regulated and influenced by the ERK/MAPK pathway in airway epithelial cells. These findings may contribute to developing novel therapeutic strategies for airway diseases.

Guia prático sobre controle ambiental para pacientes com rinite alérgica / Practical guide to environmental control for patients with allergic rhinitis

A asma e a rinite alérgica são doenças frequentes e acometem parcela significativa da população, sobretudo crianças. Frequentemente a asma e a rinite coexistem e tem sido documentado que a presença de rinite potencialmente aumenta a gravidade da asma e impacta negativamente na qualidade de vida. Entre os agentes desencadeantes/agravantes dessas doenças são apontados: aeroalérgenos (ácaros do pó domiciliar, fungos, alérgenos de baratas, epitélio de animais, polens e ocupacionais), poluentes intradomiciliares e extradomiciliares (fumaça de tabaco, material particulado liberado pela cocção/aquecimento ­ gás de cozinha, fogão a lenha) e irritantes (odores fortes, arcondicionado). O objetivo desse estudo foi identificar as medidas recomendadas para reduzir a exposição de pacientes sensíveis a esses agentes. Realizou-se busca em base de dados MEDLINE, SciELO e LILACS empregando-se os descritores: environmental control, mite, cockroach, fungi, furry pets, pollen, irritants, smoking, indoor pollution, cooking. Foram revisados os principais estudos e elaborou-se um documento em que são discutidas as relações entre exposição e aparecimento de sintomas, assim como as medidas apontadas como tendo potencial para evitar a exacerbação/ agravamento das doenças alérgicas respiratórias.

Asthma and allergic rhinitis are highly prevalent diseases and they affect a significant share of the population, especially children. Very often, asthma and rhinitis coexist, and the presence of rhinitis has been shown to potentially increase the severity of asthma, with a negative impact on quality of life. Among the triggering or aggravating agents of these conditions it is possible to list: aeroallergens (house dust mites, fungi, cockroach allergens, animal epithelium, pollens and occupational allergens), indoor and outdoor pollutants (tobacco smoke, particulate matter released by cooking/heating ­ cooking gas, wood stoves), and irritants (strong odors, air conditioning). The aim of this study was to identify measures recommended to reduce the exposure of patients sensitive to these agents. A search was conducted on the MEDLINE, SciELO, and LILACS databases, using the following keywords: environmental control, mite, cockroach, fungi, furry pets, pollen, irritants, smoking, indoor pollution, cooking. The main studies were reviewed, and a report was prepared in which the relationships between exposure and the onset of symptoms are discussed, and measures with a potential to prevent exacerbation/ aggravation of allergic respiratory diseases are presented.

Do dietary stresses affect the immune system of Periplaneta americana (Blattaria: Blattidae)?

Abstract Stresses can be caused by multiple biotic and abiotic factors and their effects can affect both the biology and the immune system of insects. American cockroach Periplaneta americana (Linnaeus, 1758) (Blattaria: Blattidae) besides being an excellent model species, has great medical importance because it can act as a mechanical vector of several pathogens. This study aimed to evaluate the influence of starvation, dehydration and both stresses on weight, and total and differential haemocyte count in P. americana adults. Each specimen was isolated in glass flasks containing or not food and/or water. They were weighed periodically. Another group received water for 24 h after the end of stress period. In the immunologic bioassay, we counted their haemocytes after the final weighing. All stresses reduced the insect weight, especially when the stresses were combined. Females of the control group gained weight and males had it unaltered. Different stress conditions and time did not influence on total haemocyte count. Insects without food and water had the proportion of prohaemocytes increased and plasmatocytes decreased. This study can serve as a basis of further studies of bioecology, behaviour and the ability of resisting insecticides, besides serving as a model to studies in other insect species.

Resumo Os estresses podem ser causados por múltiplos fatores bióticos e abióticos e seus efeitos podem afetar tanto a biologia como o sistema imune dos insetos. A barata-americana Periplaneta americana (Linnaeus, 1758) (Blattaria: Blattidae) além de ser uma excelente espécie modelo, tem grande importância médica, pois pode atuar como vetor mecânico de diversos patógenos. O objetivo desse estudo foi avaliar a influência da inanição, desidratação e ambos os estresses sobre o peso e o número total e diferencial de hemócitos em adultos de P. americana. Cada espécime foi isolado em frascos de vidro contendo ou não alimento e/ou água. Eles foram pesados periodicamente. Outro grupo recebeu água por 24 h após o término do período de estresse. Nos ensaios imunológicos, foram contados os seus hemócitos após a última pesagem. Todos os estresses reduziram o peso dos insetos, especialmente quando os estresses foram combinados. As fêmeas do grupo controle ganharam peso e os machos tiveram seu peso inalterado. As diferentes condições de estresse e tempo não influenciaram no número total de hemócitos. Os insetos sem alimento e água tiveram a proporção de pró-hemócitos aumentada e a de plasmatócitos reduzida. Esse estudo pode servir como base para estudos posteriores de bioecologia, comportamento e da habilidade de resistir aos inseticidas químicos, além de servir como modelo para estudos em outras espécies de insetos.

Pathogenicity of Aspergillus westerdijkiae to females and oothecae of Periplaneta Americana / Patogenicidade de isolado de Aspergillus westerdijkiae a fêmeas e ootecas de Periplaneta americana

ABSTRACT: Cockroach control is performed by the application of chemical insecticides which exert high selective pressure on populations and introduces synthetic substances in the environment, motivating the search for other methods of control such as entomopathogenic fungi. The objectives of this study were to investigate the pathogenicity of the JAB 42 Aspergillus westerdijkiae to females and oothecae of Periplaneta americana and to demonstrate its mechanism of action on oothecae. Suspensions containing 106 to 108 conidia/ml were used to infect females and oothecae. Mortality and other variables such as scanning electron microscopy were used to demonstrate the mechanism of action of the fungus. The isolated JAB 42 A. westerdijkiae is pathogenic to oothecae of P. americana, with low capacity to kill females. Adhesion, germination, penetration and extrusion of the fungus on the cockroach oothecae were observed.

RESUMO: O controle de baratas realizado através da aplicação de inseticidas químicos exerce alta pressão seletiva sobre as populações e introduz substâncias sintéticas no ambiente, motivando a procura por outros métodos de controle, como os fungos entomopatogênicos. Os objetivos deste trabalho foram investigar a patogenicidade do isolado JAB 42 de Aspergillus westerdijkiae a fêmeas e ootecas de Periplaneta americana e demonstrar o mecanismo de ação sobre ootecas. Suspensões contendo 106 a 108 conídios/mL do isolado foram usadas para infectar fêmeas e ootecas. A ação do fungo foi analisada pela mortalidade e outras variáveis, e por microscopia eletrônica de varredura. O isolado JAB 42 de A. westerdijkiae é patogênico a ootecas de P. americana, tendo baixa capacidade de matar fêmeas. Foi observada a adesão, germinação, penetração e extrusão do fungo sobre ootecas da barata.

New aspects about Supella longipalpa (Blattaria: Blattellidae)

The brown-banded cockroach, Supella longipalpa (Blattaria: Blattellidae) (S. longipalpa), recently has infested the buildings and hospitals in wide areas of Iran, and this review was prepared to identify current knowledge and knowledge gaps about the brown-banded cockroach. Scientific reports and peer-reviewed papers concerning S. longipalpa and relevant topics were collected and synthesized with the objective of learning more about health-related impacts and possible management of S. longipalpa in Iran. Like the German cockroach, the brown-banded cockroach is a known vector for food-borne diseases and drug resistant bacteria, contaminated by infectious disease agents, involved in human intestinal parasites and is the intermediate host of Trichospirura leptostoma and Moniliformis moniliformis. Because its habitat is widespread, distributed throughout different areas of homes and buildings, it is difficult to control. Considering its possible resistance to insecticides, the control situation may be far more complex. For improved control of S. longipalpa an integrated pest management program is needed. Sanitation, indoor insecticide spraying in the initial cockroach control phase and insecticide formulation baits are recommended simultaneously.

Cloning and purification of the first termicin-like peptide from the cockroach Eupolyphaga sinensis

Termicin is an antimicrobial peptide with six cysteines forming three disulfide bridges that was firstly isolated from the salivary glands and hemocytes of the termite Pseudacanthotermes spiniger. In contrast to many broad-spectrum antimicrobial peptides, termicin is most active against filamentous fungi. Although more than one hundred complementary DNAs (cDNAs) encoding termicin-like peptides have been reported to date, all these termicin-like peptides were obtained from Isoptera insects. Methods The cDNA was cloned by combination of cDNA library construction kit and DNA sequencing. The polypeptide was purified by gel filtration and reversed-phase high performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined by Edman degradation and mass spectrometry. Antimicrobial activity was tested against several bacterial and fungal strains. The minimum inhibitory concentration (MIC) was determined by microdilution tests. Results A novel termicin-like peptide with primary structure ACDFQQCWVTCQRQYSINFISARCNGDSCVCTFRT was purified from extracts of the cockroach Eupolyphaga sinensis (Insecta: Blattodea). The cDNA encoding Es-termicin was cloned by cDNA library screening. This cDNA encoded a 60 amino acid precursor which included a 25 amino acid signal peptide. Amino acid sequence deduced from the cDNA matched well with the result of protein Edman degradation. Susceptibility test indicated that Es-termicin showed strong ability to kill fungi with a MIC of 25 g/mL against Candida albicans ATCC 90028. It only showed limited potency to affect the growth of Gram-positive bacteria with a MIC of 200 g/mL against Enterococcus faecalis ATCC 29212. It was inactive against gram-negative bacteria at the highest concentration tested (400 g/mL). Es-termicin showed high sequence similarity with termicins from many species of termites (Insecta: Isoptera). Conclusions This is the first report of a termicin-like peptide isolated from E. sinensis that belongs to the insect order Blattodea. Our results demonstrate the diversity of termicin-like peptides, as well as antimicrobial peptides in insects.

Cloning and purification of the first termicin-like peptide from the cockroach Eupolyphaga sinensis

Abstract Background Termicin is an antimicrobial peptide with six cysteines forming three disulfide bridges that was firstly isolated from the salivary glands and hemocytes of the termite Pseudacanthotermes spiniger. In contrast to many broad-spectrum antimicrobial peptides, termicin is most active against filamentous fungi. Although more than one hundred complementary DNAs (cDNAs) encoding termicin-like peptides have been reported to date, all these termicin-like peptides were obtained from Isoptera insects. Methods The cDNA was cloned by combination of cDNA library construction kit and DNA sequencing. The polypeptide was purified by gel filtration and reversed-phase high performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined by Edman degradation and mass spectrometry. Antimicrobial activity was tested against several bacterial and fungal strains. The minimum inhibitory concentration (MIC) was determined by microdilution tests. Results A novel termicin-like peptide with primary structure ACDFQQCWVTCQRQYSINFISARCNGDSCVCTFRT was purified from extracts of the cockroach Eupolyphaga sinensis (Insecta: Blattodea). The cDNA encoding Es-termicin was cloned by cDNA library screening. This cDNA encoded a 60 amino acid precursor which included a 25 amino acid signal peptide. Amino acid sequence deduced from the cDNA matched well with the result of protein Edman degradation. Susceptibility test indicated that Es-termicin showed strong ability to kill fungi with a MIC of 25 μg/mL against Candida albicans ATCC 90028. It only showed limited potency to affect the growth of Gram-positive bacteria with a MIC of 200 μg/mL against Enterococcus faecalis ATCC 29212. It was inactive against gram-negative bacteria at the highest concentration tested (400 μg/mL). Es-termicin showed high sequence similarity with termicins from many species of termites (Insecta: Isoptera). Conclusions This is the first report of a termicin-like peptide isolated from E. sinensis that belongs to the insect order Blattodea. Our results demonstrate the diversity of termicin-like peptides, as well as antimicrobial peptides in insects.(AU)

CpG Oligodeoxynucleotide Inhibits Cockroach-Induced Asthma via Induction of IFN-gamma+ Th1 Cells or Foxp3+ Regulatory T Cells in the Lung

PURPOSE: CpG oligodeoxynucleotide (CpG-ODN), a TLR9 agonist, activates innate immunity and induces Th1 response. Although the immune modulatory effect of CpG-ODN has been extensively studied, its function in cockroach extract-induced allergic asthma has not been studied. Here, we investigated the inhibitory function of CpG-ODN in cockroach extract-induced asthma in mice with different treatment schemes. METHODS: Scheme 1: BALB/C mice were intra-nasally co-administered by cockroach extract and CpG-ODN twice a week for 3 weeks; Scheme 2: The mice were intra-nasally pre-treated with CpG-ODN at day 0 and cockroach allergen challenge was performed from day 3 as in scheme 1. Scheme 3: Cockroach allergen challenge was performed as in scheme 1 and CpG-ODN was post-treated at day 21. Then, BAL cell count, flow cytometric analysis of alveolar macrophages, regulatory T cells, and lung tissue histology, Th1 and Th2 cytokines, serum IgE, cockroach specific IgE, IgG1/IgG2a ratio, and airway hyper-responsiveness were evaluated. RESULTS: Mice with repeated intra-nasal exposure to CpG-ODN showed a dramatic decrease in eosinophilic inflammation, goblet cell hyperplasia, and airway hyper-responsiveness with reduction of IL-13, IL-5, and serum IgE, cockroach specific IgE and IgG1/IgG2a ratio. This inhibitory function might be related to the up-regulation of IL-10 and CD4+Foxp3+ regulatory T cells in the lung. Interestingly, one-time challenge of CpG-ODN either prior or posterior to cockroach extract exposure could modulate airway inflammation and hyper-responsiveness via increase of Th1 response. CONCLUSIONS: Collectively, our data suggest that CpG-ODN treatment modulates Th2 inflammation in the lung by induction of regulatory T cells or Th1 response in a cockroach-induced asthma model.

Prospecção quantitativa e qualitativa de uma nova fonte renovável de quitosana / Quantitative and qualitative exploration of a new renewable source of chitosan

A quitosana é um biopolímero funcional com grande potencial de desenvolvimento, podendo gerar diferentes tipos de materiais com variadas funções. Conforme modificações na sua estrutura, a quitosana tem encontrado aplicações nas mais diversas áreas, possuindo um grande leque de aplicações. Apesar do crescente uso da quitosana e do aumento das pesquisas por novas aplicações, a prospecção de outras opções de fontes (que não crustáceos) de quitosana não têm sido consistentemente apresentadas. O objetivo do presente projeto é realizar a prospecção quantitativa e qualitativa de uma nova fonte renovável de quitosana. Temos como uma fonte alternativa para a produção de quitosana, os blatódeos que são comumente conhecidos como baratas. Eles são organismos terrestres que apresentam uma reprodução consideravelmente rápida, se adaptam aos mais variados ambientes e tem o custo de criação baixíssimo devido à sua fácil adaptação ao ambiente e alimentação. Além disso, os blatódeos não possuem sazonalidade, e ainda realizam ecdises, podendo-se utilizar as exúvias para a produção de quitosana. Foram determinados o processo e o rendimento do processo de obtenção de quitosana a partir de blatódeos (Phoetalia pallida). Os blatódeos foram submetidos a tratamento com solução de hidróxido de sódio 50% (p/v) em temperatura de 120 ºC por sete tempos diferentes (1, 2, 3, 6, 10 e 20 horas). As quitosanas obtidas foram caracterizadas mediante técnicas de espectroscopia no Infravermelho (FTIR), comportamento térmico (TG/DTG e DSC), difração de raios-x, viscosimetria e teste de solubilidade. A obtenção de quitosana a partir de blatódeos apresentou vantagens em relação à produção a partir de crustáceos: reduzido número de etapas do processo e dispensa o tratamento com HCl, que é um poluente. O processo de obtenção de quitosana teve rendimento de aproximadamente 15%, variando de acordo com o tempo de reação. De uma maneira geral, as quitosanas de barata apresentaram características semelhantes à quitosana de camarão

Chitosan is a functional biopolymer with great development potential, which can generate different types of materials with several purposes. Depending on changes in its structure, chitosan has found applications in several areas, having a wide range of applications. Despite the increasing use of chitosan and the increase in research for new applications, the exploration of other options as sources of chitosan (other than shellfish) have not been consistently shown. The goal of this project is to conduct a quantitative and qualitative exploration of a new renewable source of chitosan. Blattaria, commonly known as cockroaches, are an alternative source for the production of chitosan. They are terrestrial organisms that present a considerably fast reproduction, adapt to many different environments and have a very low cost of growing, due to its easy adaptation to the environment and food. Moreover, the cockroaches don´t present seasonality and still perform ecdysis, where the exuvia can be used to produce chitosan. The process and the efficiency of the process of obtaining chitosan from the cockroaches, Phoetalia pallida, were determined: they were treated with a solution of sodium hydroxide 50% (w / v) at a temperature of 120 °C for seven different time periods (1, 2, 3, 6, 10 and 20 hours). Chitosans obtained therefrom were characterized by Infrared spectroscopy (FTIR), thermal behavior (TG / DTG and DSC), x-ray diffraction, viscosimetry and solubility test. Obtaining chitosan from cockroaches showed advantages over the production from shellfish: reduced number of process steps and not requiring treatment with HCl, which is a pollutant. The process of obtaining chitosan showed an efficiency of approximately 15%, depending upon the reaction time. In general, the cockroach chitosan showed characteristics similar to shrimp chitosan

Exposure to 50 Hz electromagnetic field changes the efficiency of the scorpion alpha toxin

Background Extremely low-frequency (50 Hz) electromagnetic field (ELF-EMF) is produced by electric power transmission lines and electronic devices of everyday use. Some phenomena are proposed as "first effects" of ELF-EMF: the discrete changes in the membrane potential and the increase of the calcium channel activity as well as the intracellular concentration of Ca 2+ . Interaction of the scorpion alpha toxin with the sodium channel depends on the orientation of the charges and may be perturbed by changes in the membrane polarization. The toxin induces overexcitability in the nervous system and an increase in the neurotransmitters released with different consequences, mainly the paralysis of muscles. We assumed that the exposure to ELF-EMF 0.7 mT will change the effects of the insect selective scorpion alpha toxin (recombinant LqhαIT from Leiurus quinquestriatus hebraeus) at the level of the cercal nerve function, the synaptic transmission and on the level of entire insect organism. Taking into account the compensatory mechanisms in organisms, we tested in addition ten times higher ELF-EMF on whole insects.Methods Experiments were performed in vivo on cockroaches (Periplaneta americana) and in vitro - on isolated cockroach abdominal nerve cord with cerci. In biotests, the effects of LqhαIT (10 −8 M) were estimated on the basis of the insect ability to turn back from dorsal to ventral side. Three groups were compared: the control one and the two exposed to ELF-EMF - 0.7 and 7 mT. Bioelectrical activity of the cercal nerve and of the connective nerve that leaves the terminal abdominal ganglion was recorded using extracellular electrodes. LqhαIT (5 × 10 −8 M) induced modifications of neuronal activity that were observed in the control cockroach preparations and in the ones exposed to ELF-EMF (0.7 mT). The exposure to ELF-EMF was carried out using coils with a size appropriate to the examined objects.Results The exposure to ELF-EMF (0.7 mT) modified the effects of LqhαIT (5 × 10−8 M) on activity of the cercal nerve and of the connective nerve. We observed a decrease of the toxin effect on the cercal nerve activity, but the toxic effect of LqhαIT on the connective nerve was increased. Biotests showed that toxicity of LqhαIT (10 −8 M) on cockroaches was reduced by the exposure to ELF-EMF (0.7 and 7 mT).Conclusions The exposure to 50 Hz ELF-EMF modified the mode of action of the anti-insect scorpion alpha toxin LqhαIT at cellular level of the cockroach nervous system and in biotests. Toxin appeared as a usefull tool in distinguishing between the primary and the secondary effects of ELF-EMF.(AU)

Exposure to 50 Hz electromagnetic field changes the efficiency of the scorpion alpha toxin

Background Extremely low-frequency (50 Hz) electromagnetic field (ELF-EMF) is produced by electric power transmission lines and electronic devices of everyday use. Some phenomena are proposed as first effects of ELF-EMF: the discrete changes in the membrane potential and the increase of the calcium channel activity as well as the intracellular concentration of Ca 2+ . Interaction of the scorpion alpha toxin with the sodium channel depends on the orientation of the charges and may be perturbed by changes in the membrane polarization. The toxin induces overexcitability in the nervous system and an increase in the neurotransmitters released with different consequences, mainly the paralysis of muscles. We assumed that the exposure to ELF-EMF 0.7 mT will change the effects of the insect selective scorpion alpha toxin (recombinant LqhIT from Leiurus quinquestriatus hebraeus) at the level of the cercal nerve function, the synaptic transmission and on the level of entire insect organism. Taking into account the compensatory mechanisms in organisms, we tested in addition ten times higher ELF-EMF on whole insects.Methods Experiments were performed in vivo on cockroaches (Periplaneta americana) and in vitro on isolated cockroach abdominal nerve cord with cerci. In biotests, the effects of LqhIT (10 8 M) were estimated on the basis of the insect ability to turn back from dorsal to ventral side. Three groups were compared: the control one and the two exposed to ELF-EMF 0.7 and 7 mT. Bioelectrical activity of the cercal nerve and of the connective nerve that leaves the terminal abdominal ganglion was recorded using extracellular electrodes. LqhIT (5 × 10 8 M) induced modifications of neuronal activity that were observed in the control cockroach preparations and in the ones exposed to ELF-EMF (0.7 mT). The exposure to ELF-EMF was carried out using coils with a size appropriate to the examined objects.Results The exposure to ELF-EMF (0.7 mT) modified the effects of LqhIT (5 × 108 M) on activity of the cercal nerve and of the connective nerve. We observed a decrease of the toxin effect on the cercal nerve activity, but the toxic effect of LqhIT on the connective nerve was increased. Biotests showed that toxicity of LqhIT (10 8 M) on cockroaches was reduced by the exposure to ELF-EMF (0.7 and 7 mT).Conclusions The exposure to 50 Hz ELF-EMF modified the mode of action of the anti-insect scorpion alpha toxin LqhIT at cellular level of the cockroach nervous system and in biotests. Toxin appeared as a usefull tool in distinguishing between the primary and the secondary effects of ELF-EMF.

Allergenic Characterization of a Novel Allergen, Homologous to Chymotrypsin, from German Cockroach

PURPOSE: Cockroach feces are known to be rich in IgE-reactive components. Various protease allergens were identified by proteomic analysis of German cockroach fecal extract in a previous study. In this study, we characterized a novel allergen, a chymotrypsin-like serine protease. METHODS: A cDNA sequence homologous to chymotrypsin was obtained by analysis of German cockroach expressed sequence tag (EST) clones. The recombinant chymotrypsins from the German cockroach and house dust mite (Der f 6) were expressed in Escherichia coli using the pEXP5NT/TOPO vector system, and their allergenicity was investigated by ELISA. RESULTS: The deduced amino acid sequence of German cockroach chymotrypsin showed 32.7 to 43.1% identity with mite group 3 (trypsin) and group 6 (chymotrypsin) allergens. Sera from 8 of 28 German cockroach allergy subjects (28.6%) showed IgE binding to the recombinant protein. IgE binding to the recombinant cockroach chymotrypsin was inhibited by house dust mite chymotrypsin Der f 6, while it minimally inhibited the German cockroach whole body extract. CONCLUSIONS: A novel allergen homologous to chymotrypsin was identified from the German cockroach and was cross-reactive with Der f 6.

IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2

PURPOSE: Cockroaches are the second leading allergen in Taiwan. Sensitization to Per a 2, the major American cockroach allergen, correlates with clinical severity among patients with airway allergy, but there is limited information on IgE epitopes and tissue localization of Per a 2. This study aimed to identify Per a 2 linear IgE-binding epitopes and its distribution in the body of a cockroach. METHODS: The cDNA of Per a 2 was used as a template and combined with oligonucleotide primers specific to the target areas with appropriate restriction enzyme sites. Eleven overlapping fragments of Per a 2 covering the whole allergen molecule, except 20 residues of signal peptide, were generated by PCR. Mature Per a 2 and overlapping deletion mutants were affinity-purified and assayed for IgE reactivity by immunoblotting. Three synthetic peptides comprising the B cell epitopes were evaluated by direct binding ELISA. Rabbit anti-Per a 2 antibody was used for immunohistochemistry. RESULTS: Human linear IgE-binding epitopes of Per a 2 were located at the amino acid sequences 57-86, 200-211, and 299-309. There was positive IgE binding to 10 tested Per a 2-allergic sera in 3 synthetic peptides, but none in the controls. Immunostaining revealed that Per a 2 was localized partly in the mouth and midgut of the cockroach, with the most intense staining observed in the hindgut, suggesting that the Per a 2 allergen might be excreted through the feces. CONCLUSIONS: Information on the IgE-binding epitope of Per a 2 may be used for designing more specific diagnostic and therapeutic approaches to cockroach allergy.

In-Vitro Evaluation of the Entomopathogenic Fungal Isolates of Metarhizium Anisopliae for Compatibility with Pesticides, Fungicides and Botanicals.

Isolates of the entomopathogenic fungus Metarhizium anisopliae were tested for their compatibility with insecticides, fungicides and botanical pesticides, which are being used in the field, as a prerequisite for developing as mycopesticides and their use in IPM programmes. Three concentrations (0.1X, 0.5X and 1X) of each chemical were evaluated in the laboratory based on the recommended dose for field application by food poison technique. Variation in vegetative growth and sporulation of M. anisopliae appeared to be related to the chemical nature of the formulations, its concentration and the fungal isolates in study. M19 and M48 isolates showed compatibility with imidacloprid at 0.5X and 0.1X and with fungicide sulphur at all the concentrations tested. All the four botanicals tested were found to be compatible to all the four fungal isolates and neem gold displayed maximum tolerance, at all the concentrations. M19 displayed an enhancement in the vegetative growth with imidacloprid (2%) and HIT (2-18%). 2% increase in the spore output was also recorded by M19 with chloropyrifos and sulphur. M19 and M48 isolates demonstrated compatibility with pesticides, fungicides and botanicals as well as with a cockroach management pesticide, HIT. The two isolates of M. anisopliae tested emerged as prospecting candidates for use as mycopesticide component in the combined application with pesticides like imidachloprid and fungicide, sulphur as well as botanicals in the IPM programmes.