Insect diversity in organic rice fields under two management systems of levees vegetation / Diversidade de insetos em arroz irrigado orgânico sob dois manejos da vegetação das taipas

Abstract Simplified environments characterize agroecosystems, reducing the diversity of associated plants, which are not cultivated for economic purposes, causing unbalances that can promote the emergence of cultivated plants pests, as well as the reduction of their natural enemies. Management systems that increase diversity in agroecosystems can extend the action of natural enemies of pests. Studies to understand the diversity of insects associated with rice cultivation and determine their ecological guilds can provide information about the composition and structure of such ecosystems, which can be applied to integrated pest management. Therefore, the study aimed to describe and compare groups of insects in irrigated rice fields, with organic management using two different systems of levees vegetation management, and relate them to the phenological states of rice cultivation (seedling, vegetative, and reproductive). Samples were taken in a plantation located in Águas Claras district of Viamão, RS. The total area of 18 ha was divided into two. A subarea called not cut (NC), where wild vegetation of levees was maintained, and the subarea named cut (C), where monthly cuts were made to levees vegetation, from the beginning of soil preparation until the harvest. From October 2012 to March 2013 were held weekly collections in quadrats randomly located in both the rice fields and the levees. A total of 800 insects were collected, 429 in the C subarea and 371 in the NC. There were identified 97 morphospecies in the C and 108 in NC, being 54 shared between the subareas. The captured insects were grouped into guilds: saprophages (C = 38.2%; NC = 27.5%), phytophagous (C = 28.5%; NC = 33.2%), entomophagous (grouping parasitoids and predators) (C = 29.4%; NC = 35%) and finally other insects (C = 4 %; NC = 4.3%). The peak abundance of phytophagous and entomophagous was registered in the vegetative stage of rice. At the same stage the UPGMA analysis showed that similarity in species composition was greater than 90% in the groups obtained in the paddy fields of C and NC subareas. The vegetation of levees can positively influence the presence of entomophagous in the field. Although the abundance did not change clearly, the greatest diversity in the NC areas of all the groups, may contribute to the maintenance of ecological services expanding the system resilience.

Resumo Os agroecossistemas se caracterizam por ambientes simplificados, com redução da diversidade de plantas associadas, que não são as cultivadas para fins econômicos, causando desequilíbrios que podem levar ao surgimento de insetos nocivos, assim como a diminuição de seus inimigos naturais. Sistemas de manejo que priorizem o aumento da diversidade no agroecossistema podem ampliar a ação de inimigos naturais de pragas. Estudos que busquem entender a diversidade de insetos associados ao cultivo de arroz irrigado, bem como determinar as guildas ou grupos ecológicos aos quais pertencem, podem trazer informações sobre a composição e estrutura dos ecossistemas que possam ser aplicadas no manejo integrado de pragas. Neste sentido, o estudo objetivou conhecer e comparar a diversidade de insetos entre áreas de cultivo orgânico de arroz irrigado, diferenciadas pelo manejo da vegetação das taipas e relacionar com os estádios fenológicos da cultura. As amostragens foram realizadas no distrito de Águas Claras, município de Viamão, RS. A área total de 18 ha foi subdividida em duas. Numa subárea, denominada não roçada (NR) a vegetação espontânea das taipas foi mantida, na outra, roçada (R), foram feitas roçadas mensais das taipas, desde o início do preparo do solo, até a colheita. Entre outubro de 2012 a março de 2013 realizaram-se coletas semanais, em quadrats, situados aleatoriamente tanto nas quadras de arroz quanto nas taipas. Foi coletado um total de 800 insetos, 429 na R e 371 na NR. Foram identificadas 97 morfoespécies na R e 108 na NR, das quais 54 foram compartilhadas entre as subáreas. As guildas registradas foram: saprófagos (R = 38,2%; NR = 27,5%), fitófagos (R = 28,5%; NR = 33,2%), entomófagos (reunindo parasitoides e predadores) (R = 29,4%; NR = 35%) e outros (R = 4%; NR = 4,3%). O pico de abundância de fitófagos e entomófagos foi registrado na fase vegetativa do arroz. Nesta mesma fase, a análise de UPGMA apontou que a similaridade na composição de espécies foi superior a 90% nos grupos obtidos nas lavouras das subáreas R e NR. A vegetação das taipas pode influenciar positivamente a presença de insetos entomófagos no campo. Embora a abundância não tenha variado significativamente entre as áreas, a maior diversidade na área não roçada em todos os grupos, pode contribuir na manutenção de serviços ecológicos aumentando a resiliência dos sistemas.

Insect diversity in organic rice fields under two management systems of levees vegetation.

Simplified environments characterize agroecosystems, reducing the diversity of associated plants, which are not cultivated for economic purposes, causing unbalances that can promote the emergence of cultivated plants pests, as well as the reduction of their natural enemies. Management systems that increase diversity in agroecosystems can extend the action of natural enemies of pests. Studies to understand the diversity of insects associated with rice cultivation and determine their ecological guilds can provide information about the composition and structure of such ecosystems, which can be applied to integrated pest management. Therefore, the study aimed to describe and compare groups of insects in irrigated rice fields, with organic management using two different systems of levees vegetation management, and relate them to the phenological states of rice cultivation (seedling, vegetative, and reproductive). Samples were taken in a plantation located in Águas Claras district of Viamão, RS. The total area of 18 ha was divided into two. A subarea called not cut (NC), where wild vegetation of levees was maintained, and the subarea named cut (C), where monthly cuts were made to levees vegetation, from the beginning of soil preparation until the harvest. From October 2012 to March 2013 were held weekly collections in quadrats randomly located in both the rice fields and the levees. A total of 800 insects were collected, 429 in the C subarea and 371 in the NC. There were identified 97 morphospecies in the C and 108 in NC, being 54 shared between the subareas. The captured insects were grouped into guilds: saprophages (C = 38.2%; NC = 27.5%), phytophagous (C = 28.5%; NC = 33.2%), entomophagous (grouping parasitoids and predators) (C = 29.4%; NC = 35%) and finally other insects (C = 4 %; NC = 4.3%). The peak abundance of phytophagous and entomophagous was registered in the vegetative stage of rice. At the same stage the UPGMA analysis showed that similarity in species composition was greater than 90% in the groups obtained in the paddy fields of C and NC subareas. The vegetation of levees can positively influence the presence of entomophagous in the field. Although the abundance did not change clearly, the greatest diversity in the NC areas of all the groups, may contribute to the maintenance of ecological services expanding the system resilience.

Systemic treatment with resveratrol and/or curcumin reduces the progression of experimental periodontitis in rats.

BACKGROUND AND OBJECTIVE: Periodontitis is a chronic inflammatory disease of periodontal tissues that leads to the destruction of bone and other connective tissues. Resveratrol and curcumin are plant-derived substances with biological properties that may have immunomodulatory properties. This study investigated the effect of continuous administration of resveratrol and curcumin and the association of resveratrol and curcumin on the progression of experimental periodontitis in rats. MATERIAL AND METHODS: Forty Wistar rats were assigned randomly to the following groups: group 1, experimental periodontitis + placebo (PL) (n = 10); group 2, experimental periodontitis + resveratrol (RSV) (n = 10); group 3, experimental periodontitis + curcumin (C) (n = 10); and group 4, experimental periodontitis + resveratrol + curcumin (COMBI) (n = 10). Periodontitis was induced in rats by tying a silk suture, as a ligature, around one of the first molars. Daily administration of the placebo solution, 10 mg/kg of resveratrol, 100 mg/kg of curcumin or 10 mg/kg of resveratrol plus 100 mg/kg of curcumin was carried out from day 0 to day 30. At the end of the relevant experimental periods, rats were killed and the specimens obtained were processed for morphometric analysis of bone loss. Gingival tissues surrounding the first molar were collected for quantification of interleukin (IL)-1ß, IL-4, interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) using a Luminex/MAGPIX assay. RESULTS: Intergroup comparisons of the morphometric outcomes revealed higher bone-loss values in the PL group (p < 0.05) when compared with RSV, C and COMBI groups. There was no difference in bone-loss values among RSV, C and COMBI groups (p > 0.05). The immunoenzymatic assay of the gingival tissue showed a lower concentration of IL-1ß in the COMBI group in comparison with the PL group (p < 0.05). Higher values of IL-4 were demonstrated in groups RSV, C and COMBI in comparison with the PL group (p < 0.05). Only RSV caused a reduction in the levels of IFN-γ (p < 0.05). There was no difference in the concentration of TNF-α amongst the four groups (p > 0.05). CONCLUSION: Resveratrol and curcumin are capable of reducing alveolar bone loss in an animal model of periodontitis. This occurred when these agents were added singly or in combination with one another, but there did not appear to be either synergistic or additive effects.

Influence of the vegetation management of the leeves in irrigated rice organic in diversity of Hymenoptera parasitoids / Influência do manejo da vegetação das taipas em arroz orgânico irrigado na diversidade de himenópteros parasitoides

Abstract Among the natural enemies of insect pests in rice fields, parasitoids are especially notable. To better understand the space-time dynamics of these insects, the objectives of this study were to describe and compare groups of parasitoids in organic irrigated rice fields using two management approaches for levee vegetation, and to relate them to the phenological stages of rice cultivation (the seedling, vegetative, and reproductive stages). The samples were taken in a plantation located in Viamão, RS, Brazil. The total area of 18 ha was divided into two parts: a no-cut (NC) subarea in which the wild vegetation of the levees was maintained, and a cut (C) subarea in which the levee vegetation was cut monthly. In each subarea, four Malaise traps considered as pseudo-replicas were installed and remained in the field for 24 hours at each sampling location. Collections occurred twice a month from the beginning of cultivation (October 2012) until harvest (March 2013). A total of 3,184 Hymenoptera parasitoids were collected: 2,038 individuals in the NC subarea and 1,146 in the C subarea. We identified 458 morphospecies distributed in 24 families. Mymaridae was the most abundant and Eulophidae was the richest in both subareas. A total of 198 morphospecies was shared between the subareas, including Platygastridae, Eulophidae, and Mymaridae, which were the families with the highest number of shared species. The richness and abundance of parasitoids varied according to their phenological developmental stages, with peak abundance registering during the vegetative period. The Morisita index identified three groupings, indicating a similarity that was related to the three phases of rice growth and development: seedling, vegetative and post-harvest.

Resumo Entre os inimigos naturais de insetos pragas em campos de arroz, parasitoides são especialmente notáveis. Para melhor entender a dinâmica espaço-temporal destes insetos, os objetivos desse estudo foram descrever e comparar os grupos de parasitoides em campos de arroz orgânico irrigado utilizando duas abordagens de manejo da vegetação das taipas, e relacioná-los com os estádios fenológicos da cultura (plântula, vegetativo e reprodutivo). As amostragens foram realizadas em uma plantação localizada em Viamão, RS. A área total de 18 ha foi dividida em duas partes: uma subárea não roçada (NR) na qual a vegetação espontânea das taipas foi mantida, e uma roçada (R) subárea em que a vegetação das taipas foi roçada mensalmente. Em cada subárea, quatro armadilhas Malaise consideradas pseudo-réplicas foram instaladas e mantiveram-se no campo durante 24 horas, em cada local de amostragem. As coletas ocorreram duas vezes por mês, desde o início do cultivo (outubro de 2012) até a colheita (março de 2013). Um total de 3.184 himenópteros parasitoides foram coletados: 2.038 indivíduos na subárea NR e 1.146 na R. Foram identificadas 458 morfoespécies, distribuídas em 24 famílias. Mymaridae foi a mais abundante e Eulophidae a mais rica para ambas as subáreas. Um total de 198 morfoespécies foi compartilhado entre as subáreas, incluindo Platygastridae, Eulophidae e Mymaridae, que foram as famílias com o maior número de espécies compartilhadas. A riqueza e a abundância de parasitoides variou de acordo com as fases de desenvolvimento fenológico da cultura, com pico de abundância registrado no período vegetativo. O Índice de Morisita identificou três grupamentos indicando uma similaridade relacionada às fases da cultura, plântula, vegetativo e, na pós-colheita.

Influence of the vegetation management of the leeves in irrigated rice organic in diversity of Hymenoptera parasitoids.

Among the natural enemies of insect pests in rice fields, parasitoids are especially notable. To better understand the space-time dynamics of these insects, the objectives of this study were to describe and compare groups of parasitoids in organic irrigated rice fields using two management approaches for levee vegetation, and to relate them to the phenological stages of rice cultivation (the seedling, vegetative, and reproductive stages). The samples were taken in a plantation located in Viamão, RS, Brazil. The total area of 18 ha was divided into two parts: a no-cut (NC) subarea in which the wild vegetation of the levees was maintained, and a cut (C) subarea in which the levee vegetation was cut monthly. In each subarea, four Malaise traps considered as pseudo-replicas were installed and remained in the field for 24 hours at each sampling location. Collections occurred twice a month from the beginning of cultivation (October 2012) until harvest (March 2013). A total of 3,184 Hymenoptera parasitoids were collected: 2,038 individuals in the NC subarea and 1,146 in the C subarea. We identified 458 morphospecies distributed in 24 families. Mymaridae was the most abundant and Eulophidae was the richest in both subareas. A total of 198 morphospecies was shared between the subareas, including Platygastridae, Eulophidae, and Mymaridae, which were the families with the highest number of shared species. The richness and abundance of parasitoids varied according to their phenological developmental stages, with peak abundance registering during the vegetative period. The Morisita index identified three groupings, indicating a similarity that was related to the three phases of rice growth and development: seedling, vegetative and post-harvest.

Mitochondrial DNA dynamics during in vitro culture and pluripotency induction of a bovine Rho0 cell line.

Large number of cellular changes and diseases are related to mutations in the mitochondrial DNA copy number. Cell culture in the presence of ethidium bromide is a known way of depleting mitochondrial DNA and is a useful model for studying such conditions. Interestingly, the morphology of these depleted cells resembles that of pluripotent cells, as they present larger and fragmented mitochondria with poorly developed cristae. Herein, we aimed to study the mechanisms responsible for the control of mitochondrial DNA replication during mitochondrial DNA depletion mediated by ethidium bromide and during the in vitro induction of cellular pluripotency with exogenous transcription factor expression in a bovine model. This article reports the generation of a bovine Rho0 mesenchymal cell line and describes the analysis of mitochondrial DNA copy number in a time-dependent manner. The expression of apoptosis and mitochondrial-related genes in the cells during mitochondrial DNA repletion were also analyzed. The dynamics of mitochondrial DNA during both the depletion process and in vitro reprogramming are discussed. It was possible to obtain bovine mesenchymal cells almost completely depleted of their mitochondrial DNA content (over 90%). However, the production of induced pluripotent stem cells from the transduction of both control and Rho0 bovine mesenchymal cells with human reprograming factors was not successful.

Resveratrol improves bone repair by modulation of bone morphogenetic proteins and osteopontin gene expression in rats.

This study investigated the effect of resveratrol on bone healing and its influence on the gene expression of osteogenic markers. Two calvarial defects were created and one screw-shaped titanium implant was inserted in the tibia of rats that were assigned to daily administration of placebo (control group, n=15) or 10mg/kg of resveratrol (RESV group, n=15) for 30 days. The animals were then sacrificed. One of the calvarial defects was processed for histomorphometric analysis and the tissue relative to the other was collected for mRNA quantification of bone morphogenetic protein (BMP)-2, BMP-7, osteopontin (OPN), bone sialoprotein (BSP), osteoprotegrin (OPG), and receptor activator of NF-κB ligand (RANKL). Implants were removed by applying a counter-torque force. Histomorphometric analysis revealed higher remaining defect in the calvarial defects of the control group than the RESV group (P=0.026). Resveratrol increased the counter-torque values of implant removal when compared to control therapy (P=0.031). Gene expression analysis showed a higher expression of BMP-2 (P=0.011), BMP-7 (P=0.049), and OPN (P=0.002) genes in the RESV group than in the control group. In conclusion, resveratrol improved the repair of critical-sized bone defects and the biomechanical retention of implants. Indeed, this natural agent may up-regulate the gene expression of important osteogenic markers.

Consequences of nitric oxide synthase inhibition during bovine oocyte maturation on meiosis and embryo development.

The importance of nitric oxide synthase (NOS) in bovine oocyte maturation was investigated. Oocytes were in vitro matured with the NOS inhibitor N(w)-L-nitro-arginine methyl-ester (10(-7), 10(-5) and 10(-3) m L-NAME) and metaphase II (MII) rates and embryo development and quality were assessed. The effect of L-NAME (10(-7) m) during pre-maturation and/or maturation on embryo development and quality was also assessed. L-NAME decreased MII rates (78-82%, p < 0.05) when compared with controls without L-NAME (96%). Cleavage (77-88%, p > 0.05), Day 7 blastocyst rates (34-42%, p > 0.05) and total cell numbers in blastocysts were similar for all groups (146-171 cells, p > 0.05). Day 8 blastocyst TUNEL positive cells (3-4 cells) increased with L-NAME treatment (p < 0.05). For oocytes cultured with L-NAME during pre-maturation and/or maturation, Day 8 blastocyst development (26-34%) and Day 9 hatching rates (15-22%) were similar (p > 0.05) to controls pre-matured and matured without NOS inhibition (33 and 18%, respectively), while total cell numbers (Day 9 hatched blastocysts) increased (264-324 cells, p < 0.05) when compared with the controls (191 cells). TUNEL positive cells increased when NOS was inhibited only during the maturation period (8 cells, p < 0.05) when compared with the other groups (3-4 cells). NO may be involved in meiosis progression to MII and its deficiency during maturation increases apoptosis in embryos produced in vitro. Nitric oxide synthase inhibition during pre-maturation and/or maturation affects embryo quality.

Effect of cyclin-dependent kinase (CDK) inhibition on expression, localization and activity of maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK) in bovine oocytes.

This study aimed to assess the effects of cyclin-dependent kinase (CDK) inhibition on factors involved in the control of meiosis in bovine oocytes: maturation promoting factor (MPF) (p34(cdc2) and cyclin B1) and mitogen activated protein kinase (MAPK). Oocytes were maintained at germinal vesicle (GV) stage in vitro with 10 µM of the CDK inhibitor butyrolactone I (BLI) for 24 h (inhibited). After this period, some of the oocytes were transferred to in vitro maturation (IVM) culture for 24 h (inhibited and matured). Control oocytes were assessed immediately after follicle aspiration (immature) or after in vitro maturation for 24 h (matured). Real-time PCR analyses showed that transcripts for p34(cdc2) and MAPK were detected in immature and inhibited oocytes and decreased after maturation, irrespective of CDK inhibition with BLI. Cyclin B1 was detected at similar levels in all oocyte groups. The p34(cdc2) and MAPK proteins were detected by Western blotting at similar levels in all oocyte groups, and cyclin B1 protein was detected only after maturation. Immunofluorescence detection showed that p34(cdc2) was localized in the cytoplasm and GV of immature oocytes, and then throughout the cytoplasm after maturation. Cyclin B1 and MAPK were detected in the cytoplasm in all oocyte groups. Maturation promoting factor and MAPK activities were similar throughout most of maturation for oocytes treated with or without BLI. In conclusion, CDK inhibition did not affect the expression (mRNA and protein levels) and localization of MPF and MAPK, and had nearly no effect on kinase activities during maturation.

Endothelial and inducible nitric oxide synthases in oocytes of cattle.

Nitric oxide (NO) is a chemical messenger generated by the activity of the nitric oxide synthases (NOS). The NOS/NO system appears to be involved in oocyte maturation, but there are few studies on gene expression and protein activity in oocytes of cattle. The present study aimed to investigate gene expression and protein activity of NOS in immature and in vitro matured oocytes of cattle. The influence of pre-maturation culture with butyrolactone I in NOS gene expression was also assessed. The following experiments were performed: (1) detection of the endothelial (eNOS) and inducible (iNOS) isoforms in the ovary by immunohistochemistry; (2) detection of eNOS and iNOS in the oocytes before and after in vitro maturation (IVM) by immunofluorescence; (3) eNOS and iNOS mRNA and protein in immature and in vitro matured oocytes, with or without pre-maturation, by real time PCR and Western blotting, respectively; and (4) NOS activity in immature and in vitro matured oocytes by NADPH-diaphorase. eNOS and iNOS were detected in oocytes within all follicle categories (primary, secondary and tertiary), and other compartments of the ovary and in the cytoplasm of immature and in vitro matured oocytes. Amount of mRNA for both isoforms decreased after IVM, but was maintained after pre-maturation culture. The NOS protein was detected in immature (pre-mature or not) and was still detected in similar amount after pre-maturation and maturation for both isoforms. NOS activity was detected only in part of the immature oocytes. In conclusion, isoforms of NOS (eNOS and iNOS) are present in oocytes of cattle from early folliculogenesis up to maturation; in vitro maturation influences amount of mRNA and NOS activity.

Nuclear maturation kinetics and in vitro embryo development of cattle oocytes prematured with butyrolactone I combined or not combined with roscovitine.

Cyclin dependent kinase inhibitors (CDKIs) may be used for pre-maturation culture, but can accelerate nuclear maturation. The aim of the present research was to compare the effect of butyrolactone I (BLI) alone or combined with roscovitine (ROS) at lesser than typically used concentrations on nuclear maturation kinetics and embryo development. To assess maturation kinetics (Experiment 1), oocytes were cultured in 100 microM BLI (B) or 6.25 microM BLI+12.5 microM ROS (BR) in TCM-199 for 24 h. Oocytes were subsequently submitted to in vitro maturation (IVM) in TCM-199+0.5 microg/ml FSH, 50 microg/ml LH and 10% FCS for another 24 h, during which oocytes were fixed every 3 h. In Experiment 2, oocytes were submitted to 24h pre-maturation treatments, with the inhibitors being diluted in TCM-199 or DMEM. IVM lasted 21 h in the culture media DMEM+0.5 microg/ml FSH, 50 microg/ml LH, 5% FCS and 50 ng/ml EGF. After IVM, oocytes from all groups were fertilized in vitro. Oocytes and sperm (2x10(6) sperm cells/ml) were co-cultured for 18 h. Embryos were co-cultured with granulosa cells in CR2aa for 8 days. All cultures were in droplets under oil, at 38.5 degrees C and 5% CO2 in air. In both experiments, control oocytes (C) were submitted only to IVM. In Experiment 1, at 0 h, C and B oocytes were all (100%) at the germinal vesicle stage (GV) of development. BR had fewer GV oocytes (89%, P<0.05). After 3 h IVM, B and BR had fewer oocytes in GV (84.7 and 79.6%, P>0.05) than C (100%, P<0.05). At 12 h, most oocytes were at intermediate stages (metaphase to telophase I) in all groups (approximately 80%, P>0.05). After 21 (77-89%) and 24 h (85-95%), all groups had similar metaphase II (MII) rates of development (P>0.05). In Experiment 2, cleavage (79-84%, P>0.05) and Day 7 blastocyst rates (26-36%, P>0.05) were similar. After 8 days, the group pre-matured with BR in DMEM had lesser blastocyst rates of development (32.3%) lower than C (40.1%, P<0.05). The other groups were similar to C (35-38%, P>0.05). Hatching rates were similar (10-15%, P>0.05) as were total cell numbers (141-170). In conclusion, BR is less effective in maintaining meiosis block; B and BR accelerate meiosis resumption; and use of pre-maturation medium may affect developmental rates.

Constrained total elbow arthroplasty.

In 1972, the senior author designed a condylar-sparing constrained hinge elbow prosthesis with a high-density polyethylene bushing. The condylar-sparing design allows both intercondylar and intramedullary fixation of the humeral component with methylmethacrylate cement. Reattachment of the muscles and collateral ligaments to the preserved condyles provides further stability. The clinical experience spans more than 16 years in 42 elbows with a relatively low loosening rate of 7%. The implant was removed and not replaced in three elbows: one for late infection, one for posttraumatic comminuted fracture of the distal humerus, and one for loosening of a humeral component. A series of 27 patients (31 elbows) with 24-204 months of follow-up study (average, 77 months) had excellent pain relief and an average range of motion of 129 degrees flexion, -44 degrees extension, 69 degrees pronation, and 61 degrees supination.