A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays / Uma nova abordagem de marcador tópico em cupins para bioensaios de laboratório

Abstract Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Resumo Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays / Uma nova abordagem de marcador tópico em cupins para bioensaios de laboratório

Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

A new approach to mark termites (Cornitermes cumulans (Kollar) Blattodea: Isoptera) for laboratory bioassays

Abstract Behavioral lab bioassays involving termites must be promptly performed to allow intended observations prior to death from dissecation, typical of these soft-bodied insects. To this end, topic markers have been proposed as an alternative to histological stains which, while not always toxic are inevitably lengthy to apply. Among recommended topic markers, gouache is easy to apply, dries out quickly, but it is known affect termites in the long run, being suitable only to short-term bioassays. Its alternative, colored glue, is also easy to apply, but it takes long to dry and it is too dense and heavy, being thus prone to affect termite walking patterns. Here we tested a mix of gouache and colored glue aiming to combine the qualities of both into a suitable topical marker for Cornitermes cumulans termites. Similar patterns of survival presented by marked and unmarked termites ruled out concerns about toxicity of this mixture. Such results were consistent across distinct group densities evidencing that the mixture does not interfere with, nor it is affected by, crowding effects. Because crowding regulates interindividual interactions and these underlie most behaviors, the mixture can be thought to be suitable to behavioral studies. We argue that this 1:2 glue:gouache mixture is an excellent alternative to mark termites for lab bioassays. Being atoxic, cheap, easy to apply, and non-invasive, this mixture may happen to be useful not only for termites but also in bioassaying other similarly soft-bodied insects.

Resumo Bioensaios comportamentais em laboratório com cupins devem ser realizados rapidamente a fim de garantir observações antes da morte por dissecação, típico desses insetos de corpo mole. Para este fim, marcadores tópicos têm sido propostos como uma alternativa para marcadores histológicos que, embora nem sempre tóxico, possuem uma aplicação demorada. Entre os marcadores tópicos recomendados, tinta guache é de fácil aplicação, rápida secagem, porém afeta os cupins em bioensaios longos, sendo adequado apenas para bioensaios curtos. Sua alternativa, cola colorida, também é de fácil aplicação mas leva muito tempo para secar e é muito denso e pesado, afetando os padrões de caminhamento dos cupins. No presente estudo, nós testamos uma mistura de tinta guache e cola colorida objetivando combinar as qualidades de ambos os marcadores tópicos em um marcador tópico adequado para Cornitermes cumulans. Padrões similares de sobrevivência entre cupins marcados e controle indicam a ausência de toxicidade na mistura de tinta guache e cola colorida. Tais resultados são consistentes em grupos de densidades distintas, o que comprova que a mistura não interfere, nem sofre efeitos de aglomeração. Uma vez que a aglomeração regula as interações inter-individuais e afetam a maioria dos comportamentos, a mistura pode ser adequada para estudos comportamentais. Nós argumentamos que a mistura de tinta guache e cola (1:2) é uma excelente alternativa como marcador tópico em cupins para bioensaios em laboratório. Sendo atóxico, barato, fácil de aplicar e não invasivo, esta mistura pode ser útil não só para os cupins, mas também em bioensaios com outros insetos de corpo mole.

The neglected tropical grasslands: first record of campo com murundus and its plant communities in the state of São Paulo, Brazil / Campos tropicais negligenciados: primeiro registro de campo com murundus e suas comunidades vegetais no estado de São Paulo, Brasil

Abstract Tropical grasslands have been systematically neglected worldwide in maps, conservation policies, and ecological studies. After eradicating invasive pine trees from a Cerrado reserve in southeastern Brazil, an unprecedented grassy ecosystem arose in recent satellite images. In the field, we confirmed the first record of wet grasslands with termite mounds - locally named campo com murundus ˗ beyond 21°S in the São Paulo state. Besides reporting this occurrence, we sampled the two plant communities forming this peculiar vegetation type (the mounds and the waterlogged matrix around them) to investigate if they are floristically and functionally distinct. We also explored how these two communities relate to those of the surrounding open vegetation types (savanna, dry and wet grassland). Woody plants were recorded on the mounds but not in the matrix, although the two communities share some ground layer species. Compared to the adjacent vegetation types, the mounds were floristically distinct and functionally more balanced in growth forms, dispersal syndrome, and tolerance to waterlogging. We hope this borderline record of campo com murundus can stimulate the search for other unnoticed remnants out of their known occurrence region, triggering efforts for their conservation and studies to improve comprehension of these iconic ecosystems.

Resumo Os campos naturais tropicais têm sido sistematicamente negligenciados em todo o planeta, seja nos mapas, nas políticas de conservação ou em estudos ecológicos. Após controle da invasão por Pinus em unidade de conservação do Cerrado no sudeste do Brasil, um ecossistema graminoso sem registro regional surgiu nas imagens de satélite recentes. Observação em campo resultou no primeiro registro de campo com murundus em latitude superior a 21°S, no estado de São Paulo. Além de relatar essa ocorrência, amostramos as duas comunidades vegetais que formam esse tipo de vegetação peculiar (ocorrendo sobre os murundus e na matriz alagadiça ao redor deles) para investigar se são florística e funcionalmente distintas. Também exploramos como essas duas comunidades estão relacionadas com os tipos de vegetação aberta circundantes (savana, campo seco e campo úmido). Plantas lenhosas foram registradas nos murundus, mas não na matriz, embora as duas comunidades compartilhem algumas espécies do estrato rasteiro. Em comparação com os outros tipos de vegetação adjacentes, os murundus se mostraram floristicamente distintos e funcionalmente mais equilibrados em relação às formas de crescimento, síndrome de dispersão e tolerância ao encharcamento. Esperamos que esse registro limítrofe de campo com murundus possa estimular a busca por outros remanescentes despercebidos fora da região de ocorrência já conhecida, desencadeando esforços para sua conservação e estudos que possam melhorar a compreensão desses ecossistemas icônicos.

Opportunistic predation of the Roadside Hawk (Rupornis magnirostris) on alates subterranean termites (Coptotermes testaceus) in southwestern Brazilian Amazon / Depredación oportunista del halcón caminero (Rupornis magnirostris) sobre termitas subterráneas aladas (Coptotermes testaceus) en el suroeste de la Amazonía brasileña

Abstract We report the opportunistic predation of Roadside Hawk (Rupornis magnirostris - Accipitridae) on subterranean termite (Coptotermes testaceus - Rhinotermitidae). This predatory behaviour was observed during an avifauna survey in the municipality of Candeias do Jamari, Rondônia, Brazil, in the southwestern Brazilian Amazon. There was a great abundance of C. testaceus alates because of heavy rainfall. This is the season when C. testaceus colonies produce alates in large numbers to found new colonies. The Roadside Hawk stayed on its perch and captured winged termites with little energy expenditure. The bird in question appeared to take advantage of the termite mating flight to feed opportunistically on the abundant alates. Although feeding on insects by R. magnirostris is well known, this is the first documentation of feeding on termite alates.

Resumen Informamos de la depredación oportunista de Coptotermes testaceus (Rhinotermitidae) por Rupornis magnirostris (Accipitridae). Este comportamiento depredador fue observado durante un estudio de la avifauna en el municipio de Candeias do Jamari, Rondônia, Brasil, en el suroeste de la Amazonia brasileña. Había una gran abundancia de formas aladas de C. testaceus debido a las fuertes lluvias. Es el momento en que las colonias de C. testaceus producen formas aladas en gran número para fundar nuevas colonias. El aguilucho permaneció en su percha y capturó termitas aladas con poco gasto de energía. El ave en cuestión parecía aprovechar el vuelo de apareamiento de las termitas para alimentarse de forma oportunista de las abundantes termitas aladas durante los vuelos de apareamiento. Aunque la alimentación de insectos por parte de R. magnirostris es bien conocida, ésta es la primera documentación de alimentación de termitas aladas.

Bioethanol production from urban cellulosic waste employing Alcaligenes faecalis HI-1 isolated from gut of termite Heterotermes indicola

This study assessed the potential of termite gut inhabiting bacteria towards bioconversion of cellulosic waste into biofuel. Total seven bacterial isolates from the gut of Heterotermes indicola were isolated. Among all the isolates, HI-1 produced the largest zone upon primary screening. Untreated paper had more cellulose content (73.03%) than acid (0.5%) treated paper that was used as a lignocellulosic substrate for saccharification. Among all the isolates tested, glucose yield (1.08mg/mL) was high for HI-1 isolate. Several factors were considered for optimizing augmented glucose yield (8.57mg/mL) and growth (8.07×108cfu/mL), such as temperature 37°C, pH 4.5, 5% (w/v) substrate concentration, 6 % bacterial inoculum size, agitation 150 rpm with PEG 0.25 % and Ca2+ ions 0.002 g/L. Overall 8-fold increase in glucose yield was achieved. Enzyme activity of HI-1 showed higher endoglucanase 0.29 ± 0.01 (U/mL/min) and exoglucanase 0.15±0.01 (U/mL/min) activity under optimum conditions, mentioned above. temperature 37°C, pH 4.5, substrate concentration 5%, inoculum size 6%, surfactants PEG 0.01%, ions Ca2+(0.002g/L) and agitation (120 rpm). Simultaneous saccharification and fermentation (SSF) of hydrolyzed office paper yielded 5.43mg/mL bioethanol. According to 16S rRNA sequence homology, the bacterial isolate H1 was identified as Alcaligenes faecalis. Bioethanol production from office paper untreated waste proved an effective strategy. Bacteria having natural tendency towards cellulosic waste consumption are promising for bioconversion of cellulosic waste to valuable products.

Influence of the amount of food resources in the foraging behavior of Nasutitermes corniger (Motschulsky) / Influência da quantidade de madeira no comportamento de forrageamento de Nasutitermes corniger (Motschulsky)

Food scarcity or abundance are factors regulating termites' foraging behavior in general. The aim of the present study is to evaluate the influence of four amounts of Eucalyptus grandis on foraging behavior events shown by worker and soldier of Nasutitermes corniger during laboratory tests. The tests were carried out with adult and active N. corniger colonies found in nests collected in the field, which were stored in glass cubes connected to the test arenas. Four different amounts of wood blocks were used in the tests and each amount concerned a treatment and defined a different experimental group: 1, 2, 3 and 4 blocks/arenas, with 5 repetitions. Each test lasted 60 minutes and consisted in observing, or not, the occurrence of behavioral events shown by foragers when they had contact with the treatment. The duration of each event was recorded, whenever it was observed. The number of recruited foragers and the number of workers consuming the blocks were recorded at the end of each test applied to each treatment. Nasutitermes corniger presented the three behavioral events in all treatments; however, there was not significant difference between treatments in the occurrence of the two first events, in the time taken from test start to the occurrence of a new event, in the number of recruited termites and in the number of gnawing workers. Only workers' mass recruiting was influenced by the amount of wood available. The occurrence of this event was significantly higher in treatments with greater amounts of wood. Thus, N. corniger adjusts its mass recruitment behavior in response to available food amount, which should be considered when developing baiting system for its control. (AU)

A escassez ou abundância de alimento são fatores que regulam o forrageamento do térmitas em geral. No presente estudo, avaliou-se a influência de quatro quantidades de madeira de Eucalyptus grandis em eventos comportamentais do forrageamento exibidos por operárias e soldados de Nasutitermes corniger em testes de laboratório. Os testes foram realizados com colônias maduras e ativas de N. cornigerpresentes em ninhos coletados a campo, os quais foram acondicionados em cubas de vidro conectadas a arenas testes. Blocos de madeira de mesma dimensão foram ofertados aos térmitas em quatro quantidades: 1, 2, 3 e 4 blocos/arena, com 5 repetições. Cada teste durou 60 minutos e consistiu na observação ou não da ocorrência de eventos comportamentais exibidos pelos forrageadores quando foi dado acesso a cada tratamento. Quando cada evento foi observado, o tempo de sua duração foi registrado. Ao final de cada teste registrou-se, em cada tratamento, o número de térmitas forrageadores recrutados e o número de operários consumindo os blocos. Observou-se que N. corniger exibiu os três eventos comportamentais em todos os tratamentos. Todavia, não houve diferença significativa entre os tratamentos com relação à ocorrência dos dois primeiros eventos, aos tempos transcorridos desde o início do teste até a ocorrência de cada evento, aos números de térmitas recrutados e ao número de operários em roedura. Apenas a ocorrência de recrutamento em massa dos operários foi influenciada pela quantidade de madeira ofertada, sendo que foi significativamente superior quando se ofertou os blocos de madeira na maior quantidade. Dessa forma, N. corniger ajusta seu comportamento de recrutamento em massa em resposta a quantidade de alimento disponível, o que deve ser considerado ao se desenvolver um sistema de isca para seu controle. (AU)

Termiticide activities of wood extractives of Ziziphus mauritiana (Rhamnaceae) against subterranean termites under field conditions

Abstract Synthetic wood preservatives are the causes of large-scale environmental pollution and few have been withdrawn from the commercial markets in the past years. The present studies focused on determination of naturally present extractives of Ziziphus mauritiana as less toxic wood protectant against subterranean termites. Firstly, natural resistance of Z. mauritiana heartwood against termite was determined by exposing stakes in the field. For the preparation of extractives, air-dried Z. mauritiana heartwood and bark shavings were soaked in 1 liter each of ethyl acetate, hexane, petroleum ether and water in a bottle separately. Different dipping treatment times (36 and 72 hours) at 10, 20 and 30% concentrations of extractives on Populus deltoides wooden stakes were used and stakes were exposed to termites in submerged manner. Combination of extractives in different solvents were included as separate experiment and finally, seasoning prior to extractives application on P. deltoides wooden stakes was also done and stakes were arranged in three replications for each treatment. Maximum mean percent weight loss (81.1%) was observed in case of P. deltoides followed by boiled Z. mauritiana (15.24%) in termite resistance test. Stakes treated with petroleum ether extracts had minimum weight loss alone or in combination with other extract's solvent in all experiments. Extractives in other solvents followed petroleum ether non-significantly but were significantly different from their respective control treatment, which had the highest weight loss (>60%). Transferring durability using extracts of Z. mauritiana increased resistance of non-durable P. deltoides against termites and extractives could be used as wood preservatives.

Gut anatomy of the worker caste of Neotropical genera Cylindrotermes Holmgren and Hoplotermes Light (Infraorder Isoptera, Termitidae)

ABSTRACT Studies over the last six decades have shown that the gut anatomy of the termite worker caste provides a valuable set of taxonomic characters. However, the gut anatomy of most American termitine taxa is still little known. This study investigated the anatomy of the worker gut of the Neotropical termitine genera Cylindrotermes Holmgren and Hoplotermes Light. We provide descriptions and illustrations of the gut in situ and the cuticular ornamentation of the gizzard, enteric valve, and first proctodeal segment for both genera.

A new termite species of the genus Dihoplotermes Araújo (Blattaria, Isoptera, Termitidae) from the Brazilian Amazonian rainforest / Uma nova espécie de térmita do gênero Dihoplotermes Araujo (Blattaria, Isoptera, Termitidae) da floresta Amazônica Brasileira

Dihoplotermes, a previously monotypic genus, is now composed of two species. Dihoplotermes taurus sp. nov., a new Termitinae termite species, is illustrated and described based on morphological characters from soldiers, workers, and imago. The new species is distinguished from Dihoplotermes inusitatus Araujo by having a spine-like protuberance with blunt apex on the postmentum, the presence of a spine on the gizzard and the absence of dimorphic soldiers. Furthermore, the internal morphological characteristics of a Dihoplotermes worker are described for the first time. Both internal and external morphological characters were used to improve the characterization of the species.(AU)

Dihoplotermes, que era um gênero monotípico, agora é composto por duas espécies. Dihoplotermes taurus sp. nov., uma nova espécie de cupim Termitinae, é descrita e ilustrada com base nos caracteres morfológicos dos soldados, operários e imago. A nova espécie se diferencia de Dihoplotermes inusitatus Araujo por apresentar uma protuberância em forma de espinho não pontiagudo no posmento, presença de um espinho na moela e ausência de soldados dimórficos. Além disso, são descritas pela primeira vez as características morfológicas internas de um operário de Dihoplotermes e utilizamos ambos caracteres morfológicos internos e externos para melhor caracterizar as espécies.(AU)

Effect of temperature and substrate moisture on group survival of Constrictotermes sp. (Isoptera: Termitidae) under laboratory conditions

ABSTRACT Laboratory bioassays should be performed under appropriate conditions that minimize stress and promote greater longevity to tested animals. For social insects, for instance, the stress triggered by removal individuals from nests may result in changes in their survival and behavior. Here we analyzed the effects of variations of different combinations of temperature and substrate moisture (mL of water/g of nest substrate) on group survival of Constrictotermes sp. (Termitidae: Nasutitermitinae). In general, lower substrate moisture resulted in faster mortality. On the other hand, groups survival longer at a temperature of 26 ºC and under higher substrate moisture (2-3 mL/7 g of nest substrate). The result of this study may contribute to the establishment of bioassay protocols performed with Constrictotermes sp. in the laboratory.

Evidencia molecular de la ocurrencia de bacterias celulolíticas del género Cohnella en especies neotropicales de Nasutitermitinae colectadas en el NEA / Molecular evidence that cellulolytic bacterial genus Cohnella is widespread among Neotropical Nasutitermitinae from NE Argentina

Cohnella is a highly cellulolytic bacterial genus, which can be found in a variety of habitats. The aim of this study was to assess its presence in the digestive tract of termite species collected in North-eastern Argentina: Nasutitermes aquilinus, N. corniger and Cortaritermes fulviceps. Gut homogenates were incubated with cellulosic substrate for bacterial growth. Bacterial 16S rDNA was partially amplified using new primers for Cohnella spp. and cloned. Sequences obtained showed highest similarity (97.2-99.9%) with those of Cohnella spp. previously reported from diverse environments. Phylogenetic analysis tended to group the clones according to their host species and sampling sites. These results indicate the association of Cohnella-related intestinal symbionts with three common Neotropical termites. Their potential industrial application encourages further research.

Cohnella es un género de bacterias celulolíticas que puede ser encontrado en una variedad de hábitats. El propósito de este estudio fue registrar su presencia en el tracto digestivo de termitas (Nasutitermes aquilinus, N. corniger y Cortaritermes fulviceps) colectadas en el noreste argentino (NEA). Se incubaron homogenados de intestinos en sustrato celulósico para multiplicar las bacterias. Utilizando nuevos cebadores para Cohnella spp., se amplificó una porción del ADN ribosomal 16S bacteriano, el cual fue posteriormente clonado. Las secuencias obtenidas mostraron su mayor porcentaje de similitud (97,2-99,9%) con Cohnella spp., previamente reportadas en diversos ambientes. El análisis filogenético tendió a agrupar a los clones de acuerdo a la especie hospedante y al sitio de muestreo. Estos resultados indican que especies de termitas frecuentes en el NEA albergan simbiontes intestinales relacionados con el género Cohnella. Las potenciales aplicaciones industriales de estos microorganismos animan a profundizar los estudios.

Co-expression of lignocellulase from termite and their endosymbionts / 生物工程学报

Natural lignocellulosic materials contain cellulose, hemicellulose, and lignin. Cellulose hydrolysis to glucose requires a series of lignocellulases. Recently, the research on the synergistic effect of lignocellulases has become a new research focus. Here, four lignocellulase genes encoding β-glucosidase, endo-1,4-β-glucanase, xylanase and laccase from termite and their endosymbionts were cloned into pETDuet-1 and pRSFDuet-1 and expressed in Escherichia coli. After SDS-PAGE analysis, the corresponding protein bands consistent with the theoretical values were observed and all the proteins showed enzyme activities. We used phosphoric acid swollen cellulose (PASC) as substrate to measure the synergistic effect of crude extracts of co-expressing enzymes and the mixture of single enzyme. The co-expressed enzymes increased the degradation efficiency of PASC by 44% compared with the single enzyme mixture; while the degradation rate increased by 34% and 20%, respectively when using filter paper and corn cob pretreated with phosphoric acid as substrates. The degradation efficiency of the co-expressed enzymes was higher than the total efficiency of the single enzyme mixture.

Inter and intra termites colonies comparisons of gut microbial diversity from worker and soldier caste of Globitermes sulphureus (Blattodea: Termitidae) using 16S rRNA gene

Aims: The present study aims at isolation and identifying termite gut bacteria from different Globitermes sulphureus mounds using a molecular approach based on16S rRNA genes. Methodology and results: The bacteria from the whole gut of soldier and worker castes of the termite G. sulphureus was isolated and identified. The diversity of the bacteria was compared between five different colonies in Universiti Sains Malaysia, Penang, Malaysia. The isolated bacteria were identified by using 16S rRNA gene sequencing method and subsequently used for phylogenetic analysis. Sequences analyses of identified bacteria were found to be affiliated with the phyla Firmicutes, Proteobacteria, and Actinobacteria. Soldiers and workers seemed to have little differences in bacterial species from the same colonies. We noted some bacteria which were detected in soldiers were not detected in workers. Conclusion, significance and impact of study: Differences in the culturable bacteria composition were not significant between termite colonies. However, the phylogenetic analysis revealed that most of the bacteria from one colony were slightly but distinctly different with bacteria from other colonies.

PRIMER REGISTRO DE TERMITEROS FÓSILES PARA EL PLEISTOCENO TARDÍO DE LA REGIÓN MESOPOTÁMICA (ARGENTINA): IMPLICANCIAS PALEOAMBIENTALES / First Fossil Termite Nests from the Late Pleistocene of the Mesopotamian Region (Argentina): Paleoenvironmental Implications

En esta contribución se da a conocer el primer registro de termiteros fósiles para la región Mesopotámica de Argentina, y su disposición espacial, al tiempo que se discuten sus afinidades taxonómicas e implicancias paleoambientales. Las estructuras fósiles pertenecen a nidificaciones de insectos sociales atribuibles a miembros de la Familia Termitidae, y se ubican en la zona de contacto entre la base de la Fm. Toropí/Yupoí (Pleistoceno tardío) y el tope de la Fm. Ituzaingó (Mioceno tardío-Plioceno). Se las identifica en los alrededores del Arroyo Toropí (28°36' S y 59°02' W), Departamento Bella Vista, provincia de Corrientes. El estudio de las nidificaciones y su disposición espacial se realizó en cinco campos de termiteros mediante su relevamiento y descripción morfológica. La morfología de las estructuras estudiadas coincide claramente con la de los nidos de termitas, en donde resulta característica una fuerte muralla externa. La caracterización de campo, yacencia, desarrollo, frecuencia, tamaños e interrelaciones de los termiteros fósiles y actuales sugieren una estrecha similitud con montículos del género Cornitermes. La morfología presente muestra también cierta semejanza con Tacuruichnus farinai, descripta para el Piso/Edad Marplatense (Plioceno) de la región Pampeana. Por otro lado, el análisis fitolítico indicó que las cantidades y las morfologías fitolíticas son semejantes tanto en los termiteros actuales como en los fósiles. Desde un punto de vista paleoambiental, la abundancia de estos termiteros en el contacto entre las mencionadas unidades indicaría la existencia de suelos bien desarrollados, con abundante vegetación graminosa y condiciones húmedas y cálidas con estacionalidad marcada.

In this contribution we show the first termite nests fossil record presence from the Mesopotamian region of Argentina and its spatial distribution. At the same time taxonomic affinities and paleoenvironmental implications are discussed. The discovery takes place in the contact area between the base of the Toropí / Yupoí formations (late Pleistocene) and the top of the Ituzaingó Fm. (late Miocene-Pliocene), around the Toropí stream (28°36' S y 59°02' W), Bella Vista Department, Corrientes province. These fossil structures belong to nestings of social insects, such as the termite nests type. They could be attributable to representatives of the Termitidae Family. The study of nestings and their spatial arrangement was carried out on five termite nest fields, by surveying and detailed morphological description thereof. The morphology of the studied structures clearly matches with termite nests, with a typical strong outer wall. Characterization of field development, frequency, size and relationships of extant and fossil termite nests suggest a close similarity with mounds of the Cornitermes genus. Furthermore the morphology of these fossil termite nests also shows some resemblance to Tacuruichnus farinai, described for the Marplatanse Stage (Pliocene) of the Pampean region. From a paleoenvironmental point of view, the abundance of these termite nests in the contact between these units indicates the existence of well developed soils with abundant grassy vegetation and humid warm conditions with marked seasonality.

Isolation and Characterization of Cellulase Producing Bacteria from the Gut of Termites (Odontotermes and Heterotermes Species).

Aim: To identify, characterize and compare the cellulolytic potentials of strains isolated from the gut of Odontotermes and Heterotermes species. Methodology: Termites were collected, identified, surface sterilized and used as source of cellulase producers. Enrichment of cellulose utilizers were done using liquid media containing carboxy methyl cellulose (CMC) as the sole source of carbon and their cellulolytic potentials were confirmed using congo red plate screening method. The isolates showing considerable zone of clearance were biochemically characterized. Three effective isolates were further identified by 16S rRNA gene sequence analysis. Growth curves of the strains were constructed under six different conditions (static and shaking at 25°C, 37°C and 45°C). Their cellulase activities- endoglucanase, FPase and β-glucosidase, assayed at 18 hours of incubation were compared under the above mentioned conditions. Results: Five isolates showing significant zone of clearance were selected, out of which three belonged to Bacillus and one each to Staphylococcus and Enterobacter sp. The three Bacillus sp. which were dominant cellulase producers were found to belong to Bacillus cereus (strain HT from Heterotermes sp. and ODO1, ODO2 from Odontotermes sp.). All the isolates showed high growth at 37°C under shaker condition. Bacillus cereus ODO2 displayed a higher cellulolytic potential compared to strain HT and ODO1. The endoglucanase, FPase and β-glucosidase activities of ODO2 were 5.06 U/mg, 2.52 U/mg and 6.01 U/mg respectively. ODO2 showed optimum specific activity at 37°C in shaker condition, whereas ODO1 and HT preferred static at same temperature. Conclusion: The strains obtained in the present study are potent cellulase producers and thus can find application in food, animal feed, textile, fuel and chemical industries. Optimization of media and genetic modification of the strains can further improve their efficiency. All the three isolates are promising in view of use in future.

The study progress of chemistry and pharmacology of termite nests / 国际中医中药杂志

Termite nests is rarely seen in the clinical applications of traditional Chinese medicine, generally used for the treatment of the bone injuries and internal medical diseases. According to the recent research on termite nests, this article makes a summarize of its distribution, types, pharmacognosy characteristics, chemical composition, pharmacological effects and its application research aspects, so as to provide reference for the further development and utilization of termite nests.

Insect gut microbiome - An unexploited reserve for biotechnological application

Metagenomics research has been developed over the past decade to elucidate the genomes of the uncultured microorganisms with an aim of understanding microbial ecology. On the other hand, it has also been provoked by the increasing biotechnological demands for novel enzymes, antibiotic and signal mimics. The gut microbiota of insects plays crucial roles in the growth, development and environmental adaptation to the host insects. Very recently, the insect microbiota and their genomes (microbiome), isolated from insects were recognized as a major genetic resources for bio-processing industry. Consequently, the exploitation of insect gut microbiome using metagenomic approaches will enable us to find novel biocatalysts and to develop innovative strategies for identifying smart molecules for biotechnological applications. In this review, we discuss the critical footstep in extraction and purification of metagenomic DNA from insect gut, construction of metagenomic libraries and screening procedure for novel gene identification. Recent innovations and potential applications in bioprocess industries are highlighted.

Insect gut microbiome - An unexploited reserve for biotechnological application

Metagenomics research has been developed over the past decade to elucidate the genomes of the uncultured microorganisms with an aim of understanding microbial ecology. On the other hand, it has also been provoked by the increasing biotechnological demands for novel enzymes, antibiotic and signal mimics. The gut microbiota of insects plays crucial roles in the growth, development and environmental adaptation to the host insects. Very recently, the insect microbiota and their genomes (microbiome), isolated from insects were recognized as a major genetic resources for bio-processing industry. Consequently, the exploitation of insect gut microbiome using metagenomic approaches will enable us to find novel biocatalysts and to develop innovative strategies for identifying smart molecules for biotechnological applications. In this review, we discuss the critical footstep in extraction and purification of metagenomic DNA from insect gut, construction of metagenomic libraries and screening procedure for novel gene identification. Recent innovations and potential applications in bioprocess industries are highlighted.

Caste polymorphism of apterous line of the Neotropical termite Nasutitermes corniger (Motschulsky) (Isoptera, Termitidae)

Termite societies are structured by individuals that can be grouped into castes and instars. The development of these instars in most species occurs in irregular patterns and sometimes is distinguished subcastes in physical systems that originate polymorphic soldiers and workers. In this study, we characterized the morphological diversity of castes of apterous in Nasutitermes corniger. We collected four colonies of N. corniger, one every three months between May 2011 and February 2012. Individuals of the nest were separated into groups: larval stages, workers and presoldiers and soldiers. A morphometric analysis was performed on individuals from each group based on head width, metatibia, antenna, and thorax length. The data were submitted to discriminant analysis to confirm different morphological types inside these groups. The apterous line of N. corniger is composed of one first larval instar and two second larval instar. The workers caste has two lines of development with four instars in a larger line and three instars in a lower line. Two morphological types were identified in presoldiers and soldiers. The pattern of castes was similar to other species of the genus, in which bifurcation into two lines of workers, one smaller and one larger occurs after the first molt.