Biocontrol potential of six Heterorhabditis bacteriophora strains isolated in the Azores Archipelago.

Entomopathogenic nematodes (EPNs) are closely associated with Popillia japonica and potentially used as their biological control agents, although field results proved inconsistent and evoked a continual pursuit of native EPNs more adapted to the environment. Therefore, we surveyed the Azorean Archipelago to isolate new strains of Heterorhabditis bacteriophora and to evaluate their virulence against the model organism Galleria mellonella under laboratory conditions. Six strains were obtained from pasture and coastal environments and both nematode and symbiont bacteria were molecularly identified. The bioassays revealed that Az172, Az186, and Az171 presented high virulence across the determination of a lethal dose (LD50) and short exposure time experiments with a comparable performance to Az29. After 72 hours, these virulent strains presented a mean determination of a lethal dose of 11 infective juveniles cm-2, a lethal time (LT50) of 34 hours, and achieved 40% mortality after an initial exposure time of only 60 minutes. Az170 exhibited an intermediate performance, whereas Az179 and Az180 were classified as low virulent strains. However, both strains presented the highest reproductive potential with means of 1700 infective juveniles/mg of larvae. The bioassays of the native EPNs obtained revealed that these strains hold the potential to be used in biological control initiatives targeting P. japonica because of their high virulence and locally adapted to environmental conditions.

The modulation effect of the Steinernema carpocapsae - Xenorhabdus nematophila complex on immune-related genes in Drosophila suzukii larvae.

Larvae of the invasive pest Drosophila suzukii are susceptible to the Steinernema carpocapsae - Xenorhabdus nematophila complex and an assessment of the immune-regulatory system activation in this insect was performed to understand the response to the nematode infection. The expressions of 14 immune-related genes of different pathways (Imd, Toll, Jak-STAT, ProPO, JNK, TGF-ß) were analyzed using qRT-PCR to determine variations after nematode penetration (90 min and 4 h) and after bacterial release (14 h). Before the bacteria were present, the nematodes were not recognized by the immune system of the larvae and practically none of the analyzed pathways presented variations when compared with the non-infected larvae. However, after the X. nematophila were released, PGRP-LC was activated leading to the gene upregulation of antimicrobial peptides of both the Toll and Imd pathways. Interestingly, the cellular response was inactive during the infection course as Jak/STAT and pro-phenoloxidase genes remained unresponsive to the presence of both pathogens. These results illustrate how D. suzukii immune pathways responded differently to the nematode and bacteria along the infection course.

How does the beach ecosystem change without tourists during COVID-19 lockdown?

Urban tourist beach ecosystems provide the essential service of recreation. These ecosystems also support critical ecological functions where biodiversity conservation is not usually a priority. The sudden lockdown due to the COVID-19 pandemic created a unique opportunity to evaluate the effects of human absence in these urban-coastal ecosystems. This study examined bioindicators from 29 urban tourist beaches in seven Latin-American countries and assesses their response to lockdown about some relevant anthropogenic stressors such as pollution, noise, human activities, and user density. The presence of animals and plants, as well as the intensity of stressors, were assessed through a standardized protocol during lockdown conditions. Additionally, the environmental conditions of the beaches before and during lockdown were qualitatively compared using multivariate non-parametric statistics. We found notable positive changes in biological components and a clear decrease in human stressors on almost all the beaches. Dune vegetation increased on most sites. Similarly, high burrow densities of ghost crabs were observed on beaches, except those where cleaning activity persisted. Because of the lockdown, there was an exceptionally low frequency of beach users, which in turn reduced litter, noise and unnatural odors. The observed patterns suggest that tourist beaches can be restored to natural settings relatively quickly. We propose several indicators to measure changes in beaches once lockdown is relaxed. Adequate conservation strategies will render the recreational service of tourist beaches more environmental-friendly.

Distribution patterns, carbon sources and niche partitioning in cave shrimps (Atyidae: Typhlatya).

Cave shrimps of the Typhlatya genus are common and widespread in fresh, brackish and marine groundwater throughout the Yucatan Peninsula (Mexico). These species are ideal models to test niche partitioning within sympatric species in oligotrophic systems. Nevertheless, their food sources remain unidentified, and despite their frequency and functional importance, distribution and abundance patterns of these species within caves have not been fully recognized. Here, we describe the abundance of three Typhlatya species in different temporal and spatial scales, investigate changes in water conditions, and potential sources of carbon as an indication of food origin. Species composition and abundance varied markedly in space and time revealing patterns that differed from one system to another and in relation to environmental parameters. Isotope analysis showed that each species reflects a particular δ13C and Δ14C fingerprint, suggesting they feed in different proportions from the available carbon sources. Overall, our findings suggest a niche partitioning of habitat and feeding sources amongst the three Typhlatya species investigated, where environmental characteristics and physiological differences could play an important role governing their distribution patterns.

Effect of a gradually increasing temperature on the behavioural and physiological response of juvenile Hippocampus erectus: Thermal preference, tolerance, energy balance and growth.

The physiological and behavioural responses of ectotherms to temperature is strongly dependent on the individuals' previous thermal history. Laboratory based studies investigating the mechanisms of thermoregulation in marine ectotherms, however, rarely consider key temporal elements of thermal exposure, such as the rate at which temperature changes. We tested the hypothesis that juvenile seahorses, Hippocampus erectus, from a tropical coastal lagoon in Yucatan, Mexico, would exhibit variations in physiological and behavioural descriptors of thermoregulation when submitted to contrasting regimes during 30 days: temperature constant at 25 °C (C 25); gradually increasing 1 °C every 5 days from 25 to 30 °C (GI 25-30); and constant at 30 °C (C 30). Immediately after exposure, critical maximum temperature, thermal preference, oxygen consumption, partial energy balance, growth rate and survival of seahorses were measured. Seahorses exposed to GI 25-30 showed a significantly higher critical thermal maxima (37.8 ±â€¯0.9 °C), preference (28.7 ±â€¯0.4 °C), growth (1.10 ±â€¯0.49%) and survival (97.6%) than those exposed to C 30 (36.5 ±â€¯1, 29.4 ±â€¯0.3 °C, 0.48 ±â€¯0.32%, 73.8%, respectively). Both high temperature regimes induced metabolic depression, but ramping resulted in a greater amount of energy assimilated (278.9 ±â€¯175.4 J g-1 day-1) and higher energy efficiency for growth (89.8%) than constant exposure to 30 °C (115.4 ±â€¯63.4 J g-1 day-1, 65.3%, respectively). Gradually increasing temperature allowed physiological mechanisms of thermal adjustment to take place, reflecting the capacity of juvenile H. erectus to respond to environmental change. Despite its advantage, this capacity is limited in time, since the cumulative effect of thermal exposure affected metabolic performance, eventually compromising survival. The study of seahorse response to thermal variations in the context of ocean warming needs to consider the temporal elements of thermal exposure to foresee its vulnerability under future scenarios.

Identification of chickpea seed proteins resistant to simulated in vitro human digestion.

Proteins and peptides able to resist gastrointestinal digestion and reach the intestinal mucosa have the potential to influence human health. Chickpea (Cicer arietinum L.) seed proteins are able to resist cooking (86.9% total protein) and/or in vitro simulated human digestion (15.9% total protein resists soaking, cooking and digestion with pepsin and pancreatin). To identify and characterize proteins resisting digestion we made use of different MS methodologies. The efficiency of several proteases (trypsin, AspN, chymotrypsin and LysC) was tested, and two technologies were employed (MALDI-MS/MS and LC-nESI-MS/MS). Digestion with trypsin and AspN were most successful for the identification of seed proteins. When analyzed by MALDI- MS/MS, trypsin allowed the identification of at least one protein in 60% of the polypeptide bands, while AspN allows the identification in 48%. The use of LC-nESI-MS/MS, allowed the identification of much more proteins/polypeptides from digested seeds (232 vs 17 using trypsin). The majority of the proteins found to be able to resist simulated digestion were members of the 7S vicilin and 11S legumin seed storage protein classes, which are reported to contain bio-active functions. In addition, we have found proteins that had not yet been described as potentially able to cause an impact on human health. SIGNIFICANCE: This is the first proteomic study to analyze the effect of processing and simulated human gastrointestinal digestion on the proteome of chickpea seed. Chickpea is reported to have anti-nutritional effects as well as nutraceutical properties, so the identification and characterization of the proteins able to resist digestion is crucial to understand the targets underlying such properties.

Structure of the testate amoebae community in different habitats in a neotropical floodplain.

This study evaluated the differences in composition, abundance and morphology of testate amoebae among different habitats of the same aquatic environment (plankton, aquatic macrophyte and sediment) in the Upper Paraná River floodplain. Triplicate samplings were undertaken monthly at each habitat from April 2007 to March 2008. The structure of the community of testate amoebae was different among the habitats. The species typical for each habitat, according to Indval, were classified by their shell morphology. Arcella species together with Difflugia gramen and Difflugia pseudogramem were more abundant for plankton. Trinema and Phryganella stood out by their abundance and frequency in aquatic macrophytes. Centropyxis was an indicator of sediment. The results indicated a higher frequency of hemispherical and spherical shells in plankton and spherical and elongated shells in aquatic macrophytes. In the sediment, there was a high frequency of elongated species. Our results support the hypothesis that the community of testate amoebae has different structures among the habitats, refuting the idea that the organization of this community in plankton is guided by random events like the resuspension of organisms from the sediment and their displacement from marginal vegetation.

Effects of recurrent disturbances on the periphyton community downstream of a dammed watercourse.

We evaluate experimentally the effect of a sequential disturbance-desiccation-on the structure and dynamics of a periphytic algal community in a semilotic environment of the Upper Paraná River floodplain. We tested the hypothesis that the presence of recurrent disturbances have a direct negative effect on the attributes of the periphyton community. The sequential effect of desiccation on the periphytic community promoted its significant decrease in density, while the same was not observed in species richness. When desiccation was induced in a mature community, there was no difference in the community compared to control. The sequential disturbances on the community of periphytic algae in a mature stage, was characterised by greater stability. It is believed that the effects of variation in water levels caused by upstream reservoirs can likewise also modify the structure and stability of periphytic algae in the Upper Paraná River floodplain.

Zooplankton community resilience and aquatic environmental stability on aquaculture practices: a study using net cages.

Fish farming in net cages causes changes in environmental conditions. We evaluated the resilience of zooplankton concerning this activity in Rosana Reservoir (Paranapanema River, PR-SP). Samples were taken near the net cages installed at distances upstream and downstream, before and after net cage installation. The resilience was estimated by the decrease in the groups' abundance after installing the net cages. The zooplankton community was represented by 106 species. The most abundant species were Synchaeta pectinata, S. oblonga, Conochilus coenobasis, Polyarthra dolichoptera and C. unicornis (Rotifera), Ceriodaphnia cornuta, Moina minuta, Bosmina hagmanni and C. silvestrii (Cladocera) and Notodiaptomus amazonicus (Copepoda). The resilience of microcrustaceans was affected in the growing points as this activity left the production environment for longer, delaying the natural ability of community responses. Microcrustaceans groups, mainly calanoid and cyclopoid copepods, had a different return rate. The net cage installation acted as a stress factor on the zooplankton community. Management strategies that cause fewer risks to the organisms and maximize energy flow may help in maintaining system stability.

Real-time forecasting urban drainage models: full or simplified networks?

Lead time between rainfall prediction results and flood prediction results obtained by hydraulic simulations is one of the crucial factors in the implementation of real-time flood forecasting systems. Therefore, hydraulic simulation times must be as short as possible, with sufficient spatial and temporal flood distribution modelling accuracy. One of the ways to reduce the time required to run hydraulic model simulations is increasing computational speed by simplifying the model networks. This simplification can be conducted by removing and changing some secondary elements using network simplification techniques. The emphasis of this paper is to assess how the level of urban drainage network simplification influences the computational time and overall simulation results' accuracy. The models used in this paper comprise a sewer network and an overland flow drainage system in both 1D/1D and 1D/2D approaches. The 1D/1D model is used as the reference model to generate several models with different levels of simplifications. The results presented in this paper suggest that the 1D/2D models are not yet suitable to be used in real-time flood prediction applications due to long simulation time, while on the other hand, the simplified 1D/1D models show that considerable reductions in simulation time can be achieved without compromising simulation results (flow and water depth) accuracy.

Sensitivity analysis of surface runoff generation in urban flood forecasting.

Reliable flood forecasting requires hydraulic models capable to estimate pluvial flooding fast enough in order to enable successful operational responses. Increased computational speed can be achieved by using a 1D/1D model, since 2D models are too computationally demanding. Further changes can be made by simplifying 1D network models, removing and by changing some secondary elements. The Urban Water Research Group (UWRG) of Imperial College London developed a tool that automatically analyses, quantifies and generates 1D overland flow network. The overland flow network features (ponds and flow pathways) generated by this methodology are dependent on the number of sewer network manholes and sewer inlets, as some of the overland flow pathways start at manholes (or sewer inlets) locations. Thus, if a simplified version of the sewer network has less manholes (or sewer inlets) than the original one, the overland flow network will be consequently different. This paper compares different overland flow networks generated with different levels of sewer network skeletonisation. Sensitivity analysis is carried out in one catchment area in Coimbra, Portugal, in order to evaluate overland flow network characteristics.

Biochemical study and in vitro insect immune suppression by a trypsin-like secreted protease from the nematode Steinernema carpocapsae.

A trypsin-like serine protease was purified by gel filtration and anion-exchange chromatography from the excretory-secretory products of parasitic phase Steinernema carpocapsae. The purified protease exhibited a molecular mass of about 29 kDa by SDS-PAGE and displayed a pI of 6.3. This protease exhibited high activity with trypsin-specific substrate N-Ben-Phe-Val-Arg-p-nitroanilide and was highly sensitive to aprotinin and benzamidine. The purified trypsin protease digested the chromogenic substrate N-Ben-Phe-Val-Arg-p-nitroanilide with K(m), V(max) and k(cat) values of 594.2 mum, 0.496 mum/min and 22.8/s, respectively. The optimal pH and temperature for protease activity were 9 and 30 degrees C, respectively. Internal amino acid sequencing yielded 150 amino acids and these were homologous to other trypsin sequences. In vitro investigation was carried out to monitor prophenoloxidase suppression in Galleria mellonella by the purified protease; about 38.9-52.6% suppression of prophenoloxidase was observed. The purified protease affected insect haemocyte spreading, causing cells to become spherical or round. Protease-treated actin filaments were highly disorganized in haemocytes. In vitro, G. mellonella haemocytes recognized infective juveniles of Heterorhabditis bacteriophora; however, S. carpocapsae and Steinernema glaseri were not recognized. We provide experimental evidence that the purified trypsin has the potential to alter host haemocytes, actin filaments and to inhibit host haemolymph melanization.

Biodiversity of zooplankton communities in the Upper Paraná River floodplain: interannual variation from long-term studies.

We investigated the effect of interannual variation of hydrosedimentological regime and connectivity on the zooplankton biodiversity in the Upper Paraná River floodplain. Zooplankton samplings were undertaken between 2000 and 2007, in different environments of the floodplain, including connected and isolated floodplain lakes, backwaters, rivers and channels. The zooplankton included 541 species. Rotifers showed the highest species richness and abundance. Among the zooplankton species, 71 represent new occurrence records for the floodplain. The species accumulation curve showed a continuous increase in gamma diversity, demonstrating the importance of long-term research for accurate knowledge of biodiversity in heterogeneous and dynamic ecosystems, such as the floodplains. Interannual beta diversity among studied years indicated a lesser alteration in community composition in 2001, when a long limnophase period was observed. In most of the environments, the highest species richness values were related to the greatest flooding amplitudes. Flooding amplitude, which is associated with connectivity, favors faunal exchange amongst the environments and between the pelagic and littoral zones. This explains the occurrence of both planktonic and non-planktonic species within the community. On the other hand, mean zooplankton abundance values were higher when a long isolation period occurred. Differences between the potamophase and limnophase amplitude associated with connectivity among the environments were the most important factors for the structure and dynamics of the zooplankton community in the Upper Paraná River floodplain.

Exotic species of zooplankton in the Upper Paraná River floodplain, Daphnia lumholtzi Sars, 1885 (Crustacea: Branchiopoda).

The integrity of aquatic ecosystems is being challenged worldwide by invading species, which has been one of the frequent causes of biodiversity loss. The invader may cause extinctions of vulnerable native species through predation, grazing, competition and habitat alteration. Daphnia lumholtzi G. O. Sars, 1885, a native cladoceran from Australia, Southwestern Asia and North Africa, has recently been found in the Neotropical region. The D. lumholtzi records from the Upper Paraná River floodplain were restricted to the Pombas floodplain lake (22 degrees 47' 55.92" S and 53 degrees 21' 32.58" W) and Pau Véio Backwater (22 degrees 44' 50.76' S and 53 degrees 15' 11.16' W), in 2003 and 2008, respectively. This species can be distinguished from the other Daphnia species registered in Brazil by the conspicuous pointed fornix, the sizes of the tail spine and helmet, and a carapace ventral margin with strong spines. The high temperatures in the tropical region, as well as the increase in water transparency and the decrease in nutrient concentration observed in the environments of the Upper Paraná River floodplain due to the upstream retention by dams, may favor the development of D. lumholtzi populations. The development of populations of D. lumholtzi in natural environments of the Upper Paraná River floodplain may suggest that this species is establishing in the Neotropical region.

Biodiversity of zooplankton communities in the Upper Paraná River floodplain: interannual variation from long-term studies / Biodiversidade de comunidades de zooplâncton na planície de inundação do Alto Rio Paraná: variações interanuais em estudos de longa duração

We investigated the effect of interannual variation of hydrosedimentological regime and connectivity on the zooplankton biodiversity in the Upper Paraná River floodplain. Zooplankton samplings were undertaken between 2000 and 2007, in different environments of the floodplain, including connected and isolated floodplain lakes, backwaters, rivers and channels. The zooplankton included 541 species. Rotifers showed the highest species richness and abundance. Among the zooplankton species, 71 represent new occurrence records for the floodplain. The species accumulation curve showed a continuous increase in gamma diversity, demonstrating the importance of long-term research for accurate knowledge of biodiversity in heterogeneous and dynamic ecosystems, such as the floodplains. Interannual beta diversity among studied years indicated a lesser alteration in community composition in 2001, when a long limnophase period was observed. In most of the environments, the highest species richness values were related to the greatest flooding amplitudes. Flooding amplitude, which is associated with connectivity, favors faunal exchange amongst the environments and between the pelagic and littoral zones. This explains the occurrence of both planktonic and non-planktonic species within the community. On the other hand, mean zooplankton abundance values were higher when a long isolation period occurred. Differences between the potamophase and limnophase amplitude associated with connectivity among the environments were the most important factors for the structure and dynamics of the zooplankton community in the Upper Paraná River floodplain.

O objetivo deste trabalho foi investigar o efeito da variação plurianual do regime hidrosedimentológico e da conectividade sobre a biodiversidade zooplanctônica da planície de inundação do Alto Rio Paraná. As amostragens do zooplâncton foram realizadas entre os anos de 2000 a 2007, em distintos ambientes dessa planície de inundação, incluindo lagoas abertas, lagoas fechadas, ressacos, rios e canais. A comunidade zooplanctônica foi constituída por 541 espécies, sendo os rotíferos o grupo mais especioso e abundante. Dentre essas espécies, 71 representam novas ocorrências para a planície de inundação. A curva de acumulação de espécies mostrou um incremento continuo da diversidade gama evidenciando a importância de estudos de longa duração para o conhecimento da real biodiversidade em ecossistemas heterogêneos e dinâmicos, como são as planícies de inundação. Os resultados da diversidade beta entre os anos estudados mostraram uma menor alteração da composição da comunidade em 2001, quando foi observado um longo período de limnofase. A maior riqueza média de espécies foi associada à grande amplitude de alagamento na maioria dos ambientes. A duração da inundação, associada à conectividade, favorece o intercâmbio de fauna entre os ambientes, bem como entre as regiões pelágica e litorânea, propiciando a ocorrência de espécies planctônicas e não planctônicas na comunidade. Por outro lado, a abundância da comunidade zooplanctônica foi maior quando ocorreu um extenso período de isolamento. Diferenças entre a amplitude de potamofase e a limnofase associada à conectividade entre os ambientes foram fatores preponderantes para estruturação e dinâmica da comunidade zooplanctônica na planície de Inundação do Alto Rio Paraná.

Exotic species of zooplankton in the Upper Paraná River floodplain, Daphnia lumholtzi Sars, 1885 (Crustacea: Branchiopoda) / Espécie exótica de zooplâncton na planície de inundação do Alto Rio Paraná, Daphnia lumholtzi Sars, 1885 (Crustacea: Branchiopoda)

The integrity of aquatic ecosystems is being challenged worldwide by invading species, which has been one of the frequent causes of biodiversity loss. The invader may cause extinctions of vulnerable native species through predation, grazing, competition and habitat alteration. Daphnia lumholtzi G. O. Sars, 1885, a native cladoceran from Australia, Southwestern Asia and North Africa, has recently been found in the Neotropical region. The D. lumholtzi records from the Upper Paraná River floodplain were restricted to the Pombas floodplain lake (22º 47' 55.92" S and 53º 21' 32.58" W) and Pau Véio Backwater (22º 44' 50.76" S and 53º 15' 11.16" W), in 2003 and 2008, respectively. This species can be distinguished from the other Daphnia species registered in Brazil by the conspicuous pointed fornix, the sizes of the tail spine and helmet, and a carapace ventral margin with strong spines. The high temperatures in the tropical region, as well as the increase in water transparency and the decrease in nutrient concentration observed in the environments of the Upper Paraná River floodplain due to the upstream retention by dams, may favor the development of D. lumholtzi populations. The development of populations of D. lumholtzi in natural environments of the Upper Paraná River floodplain may suggest that this species is establishing in the Neotropical region.

A integridade dos ecossistemas aquáticos está sendo desafiada no mundo inteiro por espécies invasoras, as quais tem sido uma das causas freqüentes de perda de biodiversidade. Um invasor pode causar extinções de espécies nativas vulneráveis através de predação, herbivoría, competição e alteração de habitat. Daphnia lumholtzi G. O. Sars, 1885, cladócero nativo da Austrália, sudeste da Ásia e norte da África, recentemente tem sido registrado na região neotropoical. Os registros de D. lumholtzi na planície de inundação do Alto Rio Paraná foram restritos a lagoa das Pombas (22º 47' 55.92" S e 53º 21' 32.58" O) e Ressaco do Pau Véio (22º 44' 50.76" S e 53º 15' 11.16" O), em 2003 e 2008, respectivamente. Esta espécie pode ser diferenciada das demais espécies de Daphnia encontradas no Brasil pelos conspícuos fórnices pontiagudos, tamanho dos espinhos caudal e elmo, e margem ventral com espinhos fortes. As elevadas temperaturas na região tropical, o aumento da transparência da água e a redução das concentrações de nutrientes nos ambientes da planície de inundação do alto rio Paraná, devido aos represamentos a montante, podem estar favorecendo o desenvolvimento de populações de D. lumholtzi. O desenvolvimento de populações de D. lumholtzi em ambientes naturais da planície de inundação do alto rio Paraná pode representar o estabelecimento desta espécie na região neotropical.

Bovine oocyte quality in relation to ultrastructural characteristics of zona pellucida, polyspermic penetration and developmental competence.

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus-oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP's pores were evaluated in squares of 6.4-microm width. In group B, oocytes were fertilized in vitro. After 48 h, non-cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 +/- 0.07 microm, which was smaller than the observed on oocytes of quality 2 (0.83 +/- 0.10 microm), quality 3 (1.02 +/- 0.22 microm) and quality 4 (1.38 +/- 0.59 microm) (p < or = 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p < or = 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore's diameter.

Occurrence and characterization of a nucleopolyhedrovirus from Spodoptera littoralis (Lepidoptera: Noctuidae) isolated in the azores.

A nucleopolyhedrovirus (SpliMNPV-Az) was isolated from diseased larvae of Spodoptera littoralis, collected at the Island of S. Miguel in Azores. The virulence of this isolate was tested against S. littoralis larvae in laboratory. LD50 against 2nd and 3rd instars were not significantly different, 1.44x10(4), 3.89x10(4) OBs per larvae, respectively, but both were significantly different from that against 4th instar, which was 61.3x10(4) OBs per larvae. The complete codons sequence of SpliMNPV-Az Polh gene obtained was 750 bp (NCBI GenBank Accession No. AY600451). This sequence was compared to other 38 polyhedrin genes from NPVs and to 6 granulin genes from GVs and resulted to be identical to the sequence of a SpliMNPV previously published, thus indicating that the natural host of SpliMNPV-Az must be S. littoralis. Genetic distances estimated from restriction enzymes profiles showed SpliMNPV-Az is close to the Egyptian SpliMNPV type B, despite some degree of genetic divergence suggested by slight differences observed on PstI profile.

Lesões desportivas em praticantes de atividade fisica: uma revisão bibliografica / Sport related injuries in practioners of all types of physical activity - a bibliographic report: a review

Esta revisao da literatura aborda a controvertida tematica da relacao entre atividade fisica, lesoes desportivas e saude a partir de uma perpectiva teorica que permite compreender n'ao so a natureza de cada uma desas tres categorias como tambem os tipos de associacoes estabelecidas entre elas. Conclusao: a introducao desse assunto e util a fisioterapeutas, professores de educacao fisica, dirigentes tecnicos e atletas por trazer informacoes pertinentes para ajudar a resolver problemas proprios da area de atuacao, com destaque para os males advindos dos danos desportivos ocorridos durante a pratica de atividade fisica, mesmo que o objetivo desta seja obter a saude

Purification and characterization of two distinct metalloproteases secreted by the entomopathogenic bacterium Photorhabdus sp. strain Az29.

Photorhabdus sp. strain Az29 is symbiotic with an Azorean nematode of the genus Heterorhabditis in a complex that is highly virulent to insects even at low temperatures. The virulence of the bacteria is mainly attributed to toxins and bacterial enzymes secreted during parasitism. The bacteria secrete proteases during growth, with a peak at the end of the exponential growth phase. Protease secretion was higher in cultures growing at lower temperatures. At 10 degrees C the activity was highest and remained constant for over 7 days, whereas at 23 and 28 degrees C it showed a steady decrease. Two proteases, PrtA and PrtS, that are produced in the growth medium were purified by liquid chromatography. PrtA was inhibited by 1,10-phenantroline and by EDTA and had a molecular mass of 56 kDa and an optimal activity at pH 9 and 50 degrees C. Sequences of three peptides of PrtA showed strong homologies with alkaline metalloproteases from Photorhabdus temperata K122 and Photorhabdus luminescens W14. Peptide PrtA-36 contained the residues characteristic of metzincins, known to be involved in bacterial virulence. In vitro, PrtA inhibited antibacterial factors of inoculated Lepidoptera and of cecropins A and B. PrtS had a molecular mass of 38 kDa and was inhibited by 1,10-phenanthroline but not by EDTA. Its activity ranged between 10 and 80 degrees C and was optimal at pH 7 and 50 degrees C. PrtS also destroyed insect antibacterial factors. Three fragments of PrtS showed homology with a putative metalloprotease of P. luminescens TTO1. Polyclonal antibody raised against PrtA did not recognize PrtS, showing they are distinct molecules.