Purification and characterization of a unique pectin lyase from aspergillus giganteus able to release unsaturated monogalacturonate during pectin degradation

A pectin lyase, named PLIII, was purified to homogeneity from the culture filtrate of Aspergillus giganteus grown in submerged culture containing orange peel waste as carbon source. PLIII was able to digest apple pectin and citrus pectins with different degrees of methyl esterification. Interestingly, the PLIII activity was stimulated in the presence of some divalent cations including Pb(2+) and was not significantly affected by Hg(2+). Like other pectin lyases, PLIII is stimulated by but is not dependent on Ca(2+). The main soluble product released during the degradation of pectic substances promoted by the PLIII is compatible with an unsaturated monogalacturonate. PLIII is a unique enzyme able to release unsaturated monogalacturonate as the only soluble product during the degradation of pectic substances; therefore, PLIII was classified as an exo-pectin lyase. To our knowledge, this is the first characterization of an exo-pectin lyase. The PLIII described in this work is potentially useful for ethanol production from pectin-rich biomass, besides other common applications for alkaline pectinases like preparation of textile fibers, coffee and tea fermentation, vegetable oil extraction, and the treatment of pulp in papermaking.

Anthocyanin incorporated dental copolymer: bacterial growth inhibition, mechanical properties, and compound release rates and stability by (1)h NMR

Objective. To evaluate bacterial growth inhibition, mechanical properties, and compound release rate and stability of copolymers incorporated with anthocyanin (ACY; Vaccinium macrocarpon). Methods. Resin samples were prepared (Bis-GMA/TEGDMA at 70/30 mol%) and incorporated with 2 w/w% of either ACY or chlorhexidine (CHX), except for the control group. Samples were individually immersed in a bacterial culture (Streptococcus mutans) for 24 h. Cell viability (n = 3) was assessed by counting the number of colony forming units on replica agar plates. Flexural strength (FS) and elastic modulus (E) were tested on a universal testing machine (n = 8). Compound release and chemical stability were evaluated by UV spectrophotometry and (1)H NMR (n = 3). Data were analyzed by one-way ANOVA and Tukey's test ( α = 0.05). Results. Both compounds inhibited S. mutans growth, with CHX being most effective (P < 0.05). Control resin had the lowest FS and E values, followed by ACY and CHX, with statistical difference between control and CHX groups for both mechanical properties (P < 0.05). The 24 h compound release rates were ACY: 1.33 μg/mL and CHX: 1.92 μg/mL. (1)H NMR spectra suggests that both compounds remained stable after being released in water. Conclusion. The present findings indicate that anthocyanins might be used as a natural antibacterial agent in resin based materials.

Polymeric and solid lipid nanoparticles for sustained release of carbendazim and tebuconazole in agricultural applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Trypanosoma cruzi and Its soluble antigens induce NET release by stimulating toll-like receptors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Participation of dectin-1 receptor on NETs release against Paracoccidioides brasiliensis: role on extracellular killing

Paracoccidioides brasiliensis is a dimorphic fungus from the Paracoccidioides genus, which is the causative agent of paracoccidioidomycosis, a chronic, subacute or acute mycosis, with visceral and cutaneous involvement. This disease that is acquired through inhalation primarily attacks the lungs but, can spread to other organs. Phagocytic cells as neutrophils play an important role during innate immune response against this fungus, but studies on antifungal activities of these cells are scarce. In addition to their ability to eliminate pathogens by phagocytosis and antimicrobial secretions, neutrophils can trap and kill microorganisms by release of extracellular structures composed by DNA and antimicrobial proteins, called neutrophil extracellular traps (NETs). Here, we provide evidence that P. brasiliensis virulent strain (P. brasiliensis 18) induces NETs release. These structures were well evidenced by scanning electron microscopy, and specific NETs compounds such as histone, elastase and DNA were shown by confocal microscopy. In addition, we have shown that dectin-1 receptor is the main PRR to which fungus binds to induce NETS release. Fungi were ensnared by NETs, denoting the role of these structures in confining the fungus, avoiding dissemination. NETs were also shown to be involved in fungus killing, since fungicidal activity detected before and mainly after neutrophils activation with TNF-α, IFN-γ and GM-CSF was significantly inhibited by cocultures treatment with DNAse.

Raising of horn fly haematobia irritans (L.) (diptera: muscidae) in laboratory by means of egg and larva inoculation

Several studies have required Haematobia irritans (L.) raising in laboratory. The present study assessed two methods of inoculating immature forms of H. irritans to obtain adults. In 2007, 15 Nellore steers (Bos indicus) (L.) were used for the collection of feces free of anthelmintic treatment and flies to produce for eggs and larva. For method I, 30 eggs were incubated in square filter paper (5 × 5 cm) and deposited on bovine feces (500 g) where they were kept until hatching and spontaneous penetration of larvae (L1) into the fecal mass. After 24 h, eggs were analyzed under a stereoscope microscope (40×) for the number of larvae that instinctively penetrated the feces. In method II, larvae were obtained only by natural egg hatching. At birth, 30 larvae were collected and individually inoculated, directly onto the fecal plate by employing a moistened brush. The tests were carried out at controlled temperature (28˚C ± 2˚C) and saturated humidity (80%) until the emergence of flies with both methods. The number of emerged flies was considered in the result. Using method I, 276 (76.7%) flies emerged from 360 inoculated eggs, while using method II, 283 (78.6%) flies emerged from 360 inoculated larvae. There was no significant difference (P = 0.7821) between methods for the number of flies; however, the proportion between males and females by means of larva inoculation was different from 1:1 (P = 0.0146). Results indicated that both methods led to a satisfactory production of flies and egg inoculation provided an easier establishment.

Quality of egg yolks after oxycarotenoids supplementation in hens' diets

Oxycarotenoids concentration in layer hens’ feed may be increased whenever the consumer market demands for more pigmented egg yolks. Oxycarotenoids present in Rubrivivax gelatinosus biomass grown in industrial wastewater have already proved their ability on enhancing yolk color. Moreover, there is a current interest on the antioxidant properties that some carotenoids may provide to food and health. So, in this experiment, we investigated the effects of oxycarotenoids supplementation in hens´ diets on yolk composition and stability to rancidity. Hy line hens aging 19 weeks were individually housed in wire cages equipped with feeders and drinkers. After 15 days receiving a corn basal diet nutritionally balanced, they were assigned to four different treatments, with six replicates, that lasted for 28 days: T1 – basal diet (control), T2 – basal diet + 1.5 mg/kg canthaxanthin, T3 – basal diet + 4.5 g/kg freeze dried R. gelatinosus biomass and T4 - basal diet + 4.5 g/kg spray dried R. gelatinosus biomass. Eggs laid on the last 5 days of rearing were collected; the yolks were separated, analyzed for pH and then freeze dried for the further analyses. Proximate composition was determined after drying and the rancidity (TBRAS method) was investigated at 0, 30 and 60 days of storage at room temperature/dark conditions. Yolks that received the oxycarotenoids had the lowest moisture content and the highest protein contents (P < 0.05). Lipids and pH were the same for all treatments (P > 0.05) and ashes only were higher for T2 (P < 0.05). All treatments that received oxycarotenoids had lower TBARS than control group and, among them, oxycarotenoids from spray dried R. gelatinosus biomass were the most effective to prevent rancidity (P < 0.05). So, we concluded that the use of R. gelatinosus biomass in hens feed brings positive effects to the yolk quality, since protein content is increased and conservation is increased due to decreased water content and lipid oxidation.

Controlled swollen and drug release from urea-cross-linked polyether/siloxane hybrids

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Insights into the swelling process and drug release mechanisms from cross-linked pectin/high amylose starch matrices

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biological control of the bronze bug, Thaumastocoris peregrinus, in eucalyptus plantations in Brazil

The bronze bug Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae) was detected in Brazil in 2008 and infested >180 000 ha of eucalyptus plantations in 2011. The bronze bug can cause a reduction of 10–15% in wood productivity after 2 years of heavy infestation. Although there is not an effective control method known, biological control is the main control strategy studied. An exotic egg parasitoid, Cleruchoides noackae (Hymenoptera: Mymaridae), was imported from Australia in 2012, reared in a laboratory and released in three Brazilian regions. Parasitoids were recovered at release points after 20–30 d. In 2013, preliminary evaluations demonstrated parasitoid establishment in these areas, and the parasitoid was recovered in adjacent areas after 1 year of release. Bioassays confi rmed egg parasitism of 15–20% by C. noackae. Other native natural enemies were studied. We found green lacewing Chrysoperla externa and predatory bugs Supputius cincticeps and Atopozelus opsimus preying on nymphs and/or adults of T. peregrinus. Another promising possibility is entomopathogenic fungi. Commercial formulations of Beauveria bassiana were tested with success in lab and fi eld conditions. Fusarium proliferatum and Paecilomyces cateniannulatus caused mortality of T. peregrinus in natural epizooties. After 5 years of research, it is possible to develop an integrated pest management system (IPM) for eucalyptus plantations based on biocontrol strategy for bronze bug.

Improving mass rearing techniques for Cleruchoides noackae (Hymenoptera: Mymaridae)

The bronze bug, Thaumastocoris peregrinus, is an important pest affecting Eucalyptus plantations. The egg parasitoid Cleruchoides noackae was introduced in Brazil in 2012 for biological control of this pest. A mass rearing of C. noackae was established at EMBRAPA Florestas. This paper summarizes the main techniques developed to date in order to maximize the production of adult C. noackae. The use of eggs laid on towel paper strips increased the number of emerging parasitoids when compared to cutouts of Eucalyptus leaves. Host eggs 2 days old are preferred by C. noackae over 3- or 4-day-old ovipostures. Eggs can be stored at 5 ºC for 30 days after being parasitized without signifi cant effects on parasitoid emergence; such storage is a convenient strategy. The mean parasitoid emergence varies signifi cantly with the density of host eggs; an increase in the number of host eggs offered reduced the number of parasitoids that emerged. These improvements have played a signifi cant role in the production of C. noackae that has made possible mass release of C. noackae in Brazil and the establishment of natural populations of the parasitoid, as recently confi rmed.

Resistência de genótipos de palma de óleo (Elaeis guineensis Jacq.) ao ataque de Opsiphanes invirae Hübner, 1808 (lepidoptera: nymphalidae) no Estado do Pará

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Resistência de genótipos de palma de óleo (Elaeis guineensis Jacq.) ao ataque de Opsiphanes invirae Hübner, 1808 (lepidoptera: nymphalidae) no Estado do Pará

Pós-graduação em Agronomia (Entomologia Agrícola) - FCAV

Spatial analysis of egg distribution and geographic changes in the spawning habitat of the Brazilian sardine Sardinella brasiliensis

This paper establishes the spawning habitat of the Brazilian sardine Sardinella brasiliensis and investigates the spatial variability of egg density and its relation with oceanographic conditions in the shelf of the south-east Brazil Bight (SBB). The spawning habitats of S. brasiliensis have been defined in terms of spatial models of egg density, temperature-salinity plots, quotient (Q) analysis and remote sensing data. Quotient curves (Q(C)) were constructed using the geographic distribution of egg density, temperature and salinity from samples collected during nine survey cruises between 1976 and 1993. The interannual sea surface temperature (SST) variability was determined using principal component analysis on the SST anomalies (SSTA) estimated from remote sensing data over the period between 1985 and 2007. The spatial pattern of egg occurrences in the SBB indicated that the largest concentration occurred between Paranagua and Sao Sebastiao. Spawning habitat expanded and contracted during the years, fluctuating around Paranagua. In January 1978 and January 1993, eggs were found nearly everywhere along the inner shelf of the SBB, while in January 1988 and 1991 spawning had contracted to their southernmost position. The SSTA maps for the spawning periods showed that in the case of habitat expansion (1993 only) anomalies over the SBB were zero or slightly negative, whereas for the contraction period anomalies were all positive. Sardinella brasiliensis is capable of exploring suitable spawning sites provided by the entrainment of the colder and less-saline South Atlantic Central Water onto the shelf by means of both coastal wind-driven (to the north-east of the SBB) and meander-induced (to the south-west of the SBB) upwelling.