Marine Pseudomonas putida: a potential source of antimicrobial substances against antibiotic-resistant bacteria

Bacteria isolated from marine sponges found off the coast of Rio de Janeiro, Brazil, were screened for the production of antimicrobial substances. We report a new Pseudomonas putida strain (designated P. putida Mm3) isolated from the sponge Mycale microsigmatosa that produces a powerful antimicrobial substance active against multidrug-resistant bacteria. P. putida Mm3 was identified on the basis of 16S rRNA gene sequencing and phenotypic tests. Molecular typing for Mm3 was performed by RAPD-PCR and comparison of the results to other Pseudomonas strains. Our results contribute to the search for new antimicrobial agents, an important strategy for developing alternative therapies to treat infections caused by multidrug-resistant bacteria.

Conservation and developmental expression of ubiquitin isopeptidases in Schistosoma mansoni

Several genes related to the ubiquitin (Ub)-proteasome pathway, including those coding for proteasome subunits and conjugation enzymes, are differentially expressed during the Schistosoma mansoni life cycle. Although deubiquitinating enzymes have been reported to be negative regulators of protein ubiquitination and shown to play an important role in Ub-dependent processes, little is known about their role in S. mansoni . In this study, we analysed the Ub carboxyl-terminal hydrolase (UCHs) proteins found in the database of the parasite’s genome. An in silicoana- lysis (GeneDB and MEROPS) identified three different UCH family members in the genome, Sm UCH-L3, Sm UCH-L5 and SmBAP-1 and a phylogenetic analysis confirmed the evolutionary conservation of the proteins. We performed quantitative reverse transcription-polymerase chain reaction and observed a differential expression profile for all of the investigated transcripts between the cercariae and adult worm stages. These results were corroborated by low rates of Z-Arg-Leu-Arg-Gly-Gly-AMC hydrolysis in a crude extract obtained from cercariae in parallel with high Ub conjugate levels in the same extracts. We suggest that the accumulation of ubiquitinated proteins in the cercaria and early schistosomulum stages is related to a decrease in 26S proteasome activity. Taken together, our data suggest that UCH family members contribute to regulating the activity of the Ub-proteasome system during the life cycle of this parasite.

Rediscovery of the putatively extinct ant species Simopelta minima (Brandão) (Hymenoptera, Formicidae), with a discussion on rarity and conservation status of ant species

Simopelta minima (Brandão, 1989) was originally described based on four workers collected in soil samples from a small cocoa plantation in Ilhéus, state of Bahia, northeastern Brazil. In the subsequent years after the description, this cocoa plantation was eliminated and the species was then considered extinct by the Brazilian environmental institutions. The recent rediscovery of S. minima workers in subterranean pitfall trap samples from Viçosa, state of Minas Gerais, southeastern Brazil, over 1.000 km distant from type locality, suggests that the rarity and vulnerability status of some ant species may be explained by insufficient sampling of adequate microhabitats, in time and space.

Conservation of mayflies (Insecta, Ephemeroptera) in Espírito Santo, southeastern Brazil

Conservation of mayflies (Insecta, Ephemeroptera) in Espírito Santo, southeastern Brazil. Ephemeroptera exhibits great diversity among bodies of freshwater in the Atlantic Forest, a biome that is suffering from massive human impact. Within this context, the creation of conservation units using biological information is more recommended than economic, cultural, or political criteria. The distribution pattern of 76 Ephemeroptera species was analyzed using the biogeographical methods Parsimony Analysis of Endemicity and Network Analysis Method in order to infer relevant areas for conservation of the mayfly community in Espírito Santo. The results obtained from both analyses were largely congruent, and pointed out four relevant areas for conservation: two in the south of the state, where conservation units or priority areas for conservation are well established; and two in the north, a region in the state where little conservation efforts have been historically done. Therefore, based on our analyses on mayflies, we recommend the expansion of the existing APCs or the creation of new APCs on the north of Espírito Santo.

Stichelia pelotensis (Lepidoptera, Riodinidae): conservation, notes, and rediscovery of an endangered butterfly from southern Brazil

ABSTRACT Stichelia pelotensis (Lepidoptera, Riodinidae) is an endemic and threatened butterfly from the Pampa biome in southern Brazil, and has not been recorded in its type locality in the last 56 years. Recently, a population was found in two sites from extreme south Brazil, Pelotas, Rio Grande do Sul state. These records are an important find given the conservation status of S. pelotensis, since all the information gathered is new and involve the natural history of this species. The information obtained is useful for the management, monitoring and conservation priorities of this species and its associated habitats, since its known distribution is restricted to a narrow area in the Rio Grande do Sul Coastal Plain inside this threatened biome in southern Brazil.

Basil conservation affected by cropping season, harvest time and storage period

Fresh basil (Ocimum basilicum L.) is used in food, phytotherapic industry, and in traditional therapeutic, due to its essential oil content and composition. Nevertheless basil can not be kept for long periods after harvest and its quality can be reduced. This work aimed to assess the influence of the season and harvest time in the postharvest conservation of basil stored for different periods. Basil was harvested at 8 am and 4 pm both in August/1999 and January/2000. Cuttings were conditioned in PVC packages and stored for 3, 6, and 9 days. During storage, chlorophyll content, essential oil content and composition were determined as well as microbiological analyses were carried out. Harvest season and the days of storage influenced the final content of essential oil. There was a linear decrease in the content of essential oil, in the chlorophyll content and in the number of mold and yeast colonies during storage. There was no effect of cropping season or harvest hour on essential oil composition, but the eugenol and linalool content increased during storage. Coliforms were under 0.3 MPN g-1 and the number of Staphylococcus aureus was under 1.0x10² UFC g-1.

Conservation of 'Pedro Sato' guavas under treatment with 1-methylcyclopropene

The search for techniques that extend shelf life of guava (Psidium guajava) fruits, and reduce its postharvest losses is desirable. The objective of this work was to evaluate the effects of concentrations of competitive ethylene antagonist 1-methylcyclopropene (1-MCP) on conservation of 'Pedro Sato' guava fruits. Treatments consisted of 0, 100, 300 or 900 nL L-1 of 1-MCP for 3 hours followed by storage at 25ºC, and 10ºC with 90±5% RH. The application of 900 nL L-1 1-MCP for 3 hours was efficient in delaying loss of skin color, and in keeping fruit firmness at both storage temperatures. The 300 nL L-1 of 1-MCP concentration was efficient in delaying skin color loss only when fruits were stored at 10ºC. The effect of 1-MCP was quite significant on the reduction of rot incidence at both storage temperatures. The respiratory rate was lower in fruits treated with 300 and 900 nL L-1 of 1-MCP during storage at 25ºC. The product was efficient in delaying the ripening of fruits, and the concentration of 900 nL L-1 showed the best effect.

Potassium permanganate effects in postharvest conservation of the papaya cultivar Sunrise Golden

The objective of this work was to evaluate the effects of KMnO4 on the extension of postharvest life of 'Sunrise Golden' papaya, stored under modified atmosphere and refrigeration. Fruit with up to 10% yellow peel were harvested in a commercial orchard in Linhares, state of Espírito Santo, Brazil. Sets of three fruit (unit mass of 289.9±18.5 g) were wrapped in low-density polyethylene films (28 ¼m thick) containing sachets of KMnO4 at 0, 0.5, 1, 1.5, and 2 g per bag. The bags were sealed and stored at 10.4±0.9°C and 90±5% relative humidity for 25 days. After this period, the fruit were removed from the bags and maintained at 21±0.8°C and 90±5% relative humidity until complete ripening. Four days after bag sealing, CO2 concentration stabilized in all treatments, and was higher in bags without KMnO4. In all treatments, fruit reached the climacteric respiratory peak on the third day after bag removal, coinciding with peel color index of 3.5. Increasing the KMnO4 dose reduced the losses in fruit fresh matter, consistency and pulp electrolyte leakage. Potassium permanganate was effective in maintaining the fruit at the pre-climacteric stage during the 25-day storage, and did not interfere with normal ripening after bag removal.

In vitro conservation of Piper aduncum and Piper hispidinervum under slow-growth conditions

The objective of this work was to evaluate in vitro storage of Piper aduncum and P. hispidinervum under slow-growth conditions. Shoots were stored at low temperatures (10, 20 and 25°C), and the culture medium was supplemented with osmotic agents (sucrose and mannitol - at 1, 2 and 3%) and abiscisic acid - ABA (0, 0.5, 1.0, 2.0 and 3.0 mg L-1). After six-months of storage, shoots were evaluated for survival and regrowth. Low temperature at 20ºC was effective for the in vitro conservation of P. aduncum and P. hispidinervum shoots. In vitro cultures maintained at 20ºC on MS medium showed 100% survival with slow-growth shoots. The presence of mannitol or ABA, in the culture medium, negatively affected shoot growth, which is evidenced by the low rate of recovered shoots.

Genetic variability of marine shrimp in the Brazilian industry

The objective of this work was to estimate the genetic variability level and distribution in Brazilian broodstocks of marine shrimp (Litopenaeus vannamei). Nine of the country's largest hatcheries were evaluated using codominant and highly polymorphic microsatellite markers. The results obtained from genotyping of ten microsatellite loci are indicative of genetic variability that is compatible with that found in wild populations of L. vannamei in Mexico and Central America. A possible explanation is the highly diversified and relatively recent origin of the available broodstocks. Bayesian analysis detected a signal for five founding populations. The distribution of genetic distances partially reflects geographical location, and this information will be useful for the creation of new broodstocks. Therefore, L. vannamei genetic variability among nine of the largest national hatcheries can be considered high.

Growth of Crassostrea gasar cultured in marine and estuary environments in Brazilian waters

The objective of this work was to evaluate the growth of the mangrove oyster Crassostrea gasar cultured in marine and estuarine environments. Oysters were cultured for 11 months in a longline system in two study sites - São Francisco do Sul and Florianópolis -, in the state of Santa Catarina, Southern Brazil. Water chlorophyll-α concentration, temperature, and salinity were measured weekly. The oysters were measured monthly (shell size and weight gain) to assess growth. At the end of the culture period, the average wet flesh weight, dry flesh weight, and shell weight were determined, as well as the distribution of oysters per size class. Six nonlinear models (logistic, exponential, Gompertz, Brody, Richards, and Von Bertalanffy) were adjusted to the oyster growth data set. Final mean shell sizes were higher in São Francisco do Sul than in Florianópolis. In addition, oysters cultured in São Francisco do Sul were more uniformly distributed in the four size classes than those cultured in Florianópolis. The highest average values of wet flesh weight and shell weight were observed in São Francisco do Sul, whereas dry flesh weight did not differ between the sites. The estuary environment is more promising for the cultivation of oysters.

In vitro selection of bacteria with potential for use as probiotics in marine shrimp culture

The objective of this work was to isolate strains of lactic acid bacteria with probiotic potential from the digestive tract of marine shrimp (Litopenaeus vannamei), and to carry out in vitro selection based on multiple characters. The ideotype (ideal proposed strain) was defined by the highest averages for the traits maximum growth velocity, final count of viable cells, and inhibition halo against nine freshwater and marine pathogens, and by the lowest averages for the traits duplication time and resistance of strains to NaCl (1.5 and 3%), pH (6, 8, and 9), and biliary salts (5%). Mahalanobis distance (D²) was estimated among the evaluated strains, and the best ones were those with the shortest distances to the ideotype. Ten bacterial strains were isolated and biochemically identified as Lactobacillus plantarum (3), L. brevis (3), Weissella confusa (2), Lactococcus lactis (1), and L. delbrueckii (1). Lactobacillus plantarum strains showed a wide spectrum of action and the largest inhibition halos against pathogens, both Gram-positive and negative, high growth rate, and tolerance to all evaluated parameters. In relation to ideotype, L. plantarum showed the lowest Mahalanobis (D²) distance, followed by the strains of W. confusa, L. brevis, L. lactis, and L. delbrueckii. Among the analyzed bacterial strains, those of Lactobacillus plantarum have the greatest potential for use as a probiotic for marine shrimp.

Effects of biofilm and refrigeration on acerola postharvest conservation

The objective of this work was to compare the effects of four different concentrations of cassava starch film and storage temperature on shelf life and the quality of a genotype of acerola from the Germplasm Active Bank (GAB) of Federal Rural University of Pernambuco, Brazil. Sound orange-reddish acerola fruits were washed with a chlorine solution (100 mg.L-1 active chlorine) and randomly distributed into different lots. The fruits were dipped for 3 min in a cassava starch suspension with concentrations of 1, 2, 3 and 4% (w/v) and the control without coating, stored at 10ºC (85% RH) and 22ºC (85% RH). The total soluble solids (TSS), pH, titratable acidity (TA) and ascorbic acid (AA) were determined at harvest and regular interval during storage. The use of cassava biofilm at 1% on acerola fruits maintained the highest ascorbic acid content and the temperature of 10ºC extended storage life. The fruits coated with 1 and 2% biofilm could be stored for a period up to 15 days at 10ºC, with acceptable quality characteristics.

Conservation of Campomanesia adamantium (CAMB.) O. berg seeds in different packaging and at varied temperatures

This article aims at evaluating the effects of different packaging and varied storage temperatures on the germination potential of seeds of Campomanesia adamantium Camb. O. Berg. The seeds were packaged in glass, aluminum foil and plastic containers, or maintained inside intact fruits at 5, 10 and 15 ºC during 0, 7, 14 and 21 days. After these periods the seeds were sown in Germitest® germination paper and maintained in incubation chambers at 25 ºC under constant white light for 42 days. Seed moisture contents were evaluated both before and after storage, as well as germination percentages, germination speed index, root and aerial portion of seedlings lengths, and total dry weights. All possible combinations of packing materials, temperatures and storage times were tested, with four repetitions of 25 seeds for each treatment. C. adamantium seeds showed initial water contents of 31.5%. Glass and aluminum packaging were efficient at maintaining the water content of the seeds, and provided greater germination speed index than the other packaging materials. Germination percentages, seedlings lengths and dry weights did not vary among the different temperatures tested. C. adamantium seeds can be stored for up to 21 days at temperatures between 5 and 15 ºC without altering their physiological quality. In terms of cost-benefit efficiencies, these seeds can be stored without significant damage for 21 days while still inside the fruits at temperatures of 5, 10 or 15 ºC.