Extraction and characterization of volatile compounds in Maytenus ilicifolia, using high-pressure CO2

This work is focused on the chemical characterization of Maytenus ilicifolia extracts obtained from high-pressure CO2 extraction. The experiments were performed in a laboratory-scale unit, where the effect of particle size (35-200 mesh), CO2 mass flow rate (1-3 g/min), temperature (293-323 K), pressure (100-250 bar), and extraction time was investigated in terms of liquid yield and chemical composition of the extracts. Results show that the particle size and CO2 mass flow rate did not affect the liquid yield, whereas the extraction temperature and solvent density exerted a pronounced effect on both liquid yield and chemical distribution of volatile compounds. The extracts were chemically analyzed with regard to phytol, squalene, vitamin E, stigmasterol, friedelan-3-ol, friedelin, dodecanoic acid and geranyl acetate in a GC/MSD.     

Coffee shade with Mimosa scabrella Benth. for frost protection in southern Brazil

Damage by radiative frosts is a major limiting factor for coffee cultivation in southern Brazil (south of 20° S latitude). The use of Mimosa scabrella (bracatinga) as a shade tree, to modify the local energy balance and thus prevent damage to the coffee plants, has been evaluated from 1986 to 1994. The study was carried out near Londrina, Parana State (23°23' S, 51°11′ W). During the experimental period, several radiative frosts with intensity ranging from moderate to very severe occurred at the site. Minimum coffee leaf temperatures during these events were 2 to 4 °C higher in the shaded plots. Due to frost protection, coffee bean yields on the average of 7 harvests were higher on the shaded plots. The potential of this system for frost protection in southern Brazil is discussed.     

Organic carbon pools in a Luvisol under agroforestry and conventional farming systems in the semi-arid region of Ceará, Brazil

Many environmental benefits have been attributed to agroforestry systems in various ecosystems around the world. However, there is a limited amount of information to evaluate this agricultural system in the semi-arid region, specifically in the region of Ceará, Brazil. The objective of this work was to evaluate five agricultural (four agroforestry and one conventional) systems in order to test the hypothesis that the agroforestry systems promote an increase in the soil organic carbon stocks and organic carbon pools, thus improving soil quality. The following treatments were tested: agrosilvopasture (AGP), silvopasture (SILV), traditional agroforestry (TRAG), intensive cropping (IC), and native forest (NF). The soil samples were collected at four depths: 0–6, 6–12, 12–20 and 20–40 cm. Total soil organic carbon stocks and the organic carbon pools (microbial biomass-C, mineralizable-C, oxidizable-C, free, occluded light fraction organic matter, and C in the humic substances) were analyzed. After 5 years of experimental cultivation, the soil under the SILV system presented the best results for the attributes studied, preserving, and in some cases, improving these attributes, when compared to the other conditions. The traditional agroforestry system (TRAG) reduced total organic carbon stocks and, consequently, C in some organic matter compartments, indicating that the fallow period was not sufficient to maintain soil quality. The AGP and IC systems presented significant losses in some of the soil organic matter (SOM) pools, suggesting that the soil environment had been degraded. The most labile SOM components were considered sensitive indicators of change in the soil quality. The silvopasture system can, therefore, be recommended as an alternative soil management strategy for food production and for the maintenance of soil quality and agricultural sustainability in the semiarid region of Ceará state.     

Inhibition of photophosphorylation and electron transport chain in thylakoids by lasiodiplodin, a natural product from Botryosphaeria rhodina

Four natural products were isolated from the fungus Botryosphaeria rhodina, and their effects on photosynthesis were tested. Only lasiodiplodin (1) inhibited ATP synthesis and electron flow from water to methylviologen; therefore, it acts as a Hill reaction inhibitor in freshly lysed spinach thylakoids. Photosystem I and II and partial reactions as well as ATPase were measured in the presence of 1. Three new different sites of 1 interaction and inhibition were found: one at CF1, the second in the water-splitting enzyme, and the third at the electron-transfer path between P680 and QA; these targets are different from that of the synthetic herbicides present. Electron transport chain inhibition by 1 was corroborated by fluorescence induction kinetics studies.     

Pedigree analysis in the Brazilian Zebu breeds

Zebu breeds play an important role in cattle production systems in Brazil. To assess the genetic variability from animals in the Herd Books of Nelore, Gir and Guzerat breeds, generation intervals, inbreeding, effective population size and parameters of gene origin (effective number of founders, ancestors and founder genomes) were calculated using pedigree records from 1938 to 1998. Breed subdivision was quantified by Wright's F -statistics. Calculations were separately carried out for consecutive 4-year intervals in the period 1979–98. Generation interval was around 8 years for the three breeds. Total inbreeding increased in all the breeds reaching values of 2.13%, 2.28% and 1.75%. Effective population size decreased from 85 to 68 in Nelore, from 70 to 45 in Gir and remained nearly constant around 104 in Guzerat. The quantities assessing the number of contributing ancestors decreased with time in all the breeds, and in the last analysed period the most important ancestor accounted for 14%, 3.1% and 4.1% in Nelore, Gir and Guzerat, respectively. Results indicate that the studied breeds are suffering from a loss of genetic variability which can result in negative effects on breeding and conservation purposes.     

Use of a Low Dose of Equine Purified FSH to Induce Multiple Ovulations in Mares

The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19°45′45′S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post‐ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35‐mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35‐ to 40‐mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively – one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low‐dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.     

Influence of different levels of concentrate and ruminally undegraded protein on digestive variables in beef heifers

This experiment evaluated the effect of 2 levels of diet concentrate (20 and 40% of DM) and 2 levels of ruminally undegraded protein (RUP: 25 and 40% of CP) on nutrient intake, total and partial apparent nutrient digestibility, microbial protein synthesis, and ruminal and physiological variables. Eight Nellore heifers (233 +/- 14 kg of BW) fitted with ruminal, abomasal, and ileal cannulas were used. The animals were held in individual sheltered pens of approximately 15 m(2) and fed twice daily at 0800 and 1600 h for ad libitum intake. Heifers were allocated in two 4 x 4 Latin square designs, containing 8 heifers, 4 experimental periods, and 4 treatments in a 2 x 2 factorial arrangement. All statistical analyses were performed using PROC MIXED of SAS. Titanium dioxide (TiO(2)) and chromic oxide (Cr(2)O(3)) were used to estimate digesta fluxes and fecal excretion. Purine derivative (PD) excretion and abomasal purine bases were used to estimate the microbial N (MN) synthesis. No significant interaction (P > 0.10) between dietary levels of RUP and concentrate was observed. There was no effect of treatment (P = 0.24) on DMI. Both markers led to the same estimates of fecal, abomasal, and ileal DM fluxes, and digestibilities of DM and individual nutrients. Ruminal pH was affected by sampling time (P < 0.001), but no interaction between treatment and sampling time was observed (P = 0.71). There was an interaction between treatment and sampling time (P < 0.001) for ruminal NH(3)-N concentration. A linear decrease (P = 0.04) over sampling time was observed for the higher level of RUP, whereas a quadratic effect (P < 0.001) of sampling time was observed for the lower level of RUP. The higher level of dietary concentrate led to greater MN yield regardless of the level of RUP. The MN yield and the efficiency of microbial yield estimated from urinary PD excretion produced greater (P < 0.01) values than those estimated by either TiO(2) or Cr(2)O(3), which did not differ (P = 0.63) from each other. However, all methods yielded values that were within the range reported in the literature. In conclusion, no interactions between dietary levels of RUP and concentrate were observed for ruminal and digestive parameters. Neither RUP nor concentrate level affected DMI. Titanium dioxide showed to be similar to Cr(2)O(3) as an external marker to measure digestibility and nutrient fluxes in cattle.     

Immunogenicity in dogs of three recombinant antigens (TSA, LeIF and LmSTI1) potential vaccine candidates for canine visceral leishmaniasis

Control of canine visceral leishmaniasis (VL) remains a difficult and serious problem mostly because there is no reliable and effective vaccine available to prevent this disease. A mixture of three recombinant leishmanial antigens (TSA, LeIF and LmSTI1) encoded by three genes highly conserved in the Leishmania genus have been shown to induce excellent protection against infection in both murine and simian models of cutaneous leishmaniasis. A human clinical trial with these antigens is currently underway. Because of the high degree of conservation, these antigens might be useful vaccine candidates for VL as well. In the present study, using the dog model of the visceral disease, we evaluated the immunogenicity of these three antigens formulated with two different adjuvants, MPL-SE and AdjuPrime. The results were compared with a whole parasite vaccine formulated with BCG as the adjuvant. In order to investigate if sensitization with the recombinant antigens would result in recognition of the corresponding native parasite antigens upon infection, the animals were exposed for four weeks after the termination of the immunization protocol with the recombinant antigens to a low number of L. chagasi promastigotes, an etiological agent of VL. Immune response was evaluated by quantitative ELISA in the animal sera before and after exposure to the viable parasites. Both antigen specific IgG1 and IgG2 antibody levels were measured. Immunization of dogs with the recombinant antigens formulated in either MPL-SE or AdjuPrime resulted in high antibody levels particularly to LmSTI1. In addition, this immunization although to low levels, resulted in the development of antibody response to the whole parasite lysate. Importantly, experimental exposure with low numbers of culture forms of L. chagasi promastigotes caused a clear boost in the immune response to both the recombinant antigens and the corresponding native molecules. The boost response was predominantly of the IgG2 isotype in animals primed with the recombinant antigens plus MPL-SE. In contrast, animals primed with the recombinant antigens formulated in AdjuPrime as well as animals vaccinated with crude antigen preparation responded with mixed IgG1/IgG2 isotypes. These results point to the possible use of this antigen cocktail formulated with the adjuvant MPL-SE in efficacy field trials against canine VL.     

Effects of bean line pattern mosaic virus on the monocyclic components of rust and angular leaf spot of Phaseolus bean at different temperatures

Monocyclic components (development rate during the incubation period or latent period, lesion density, lesion size and disease severity) of rust ( Uromyces appendiculatus ) and of angular leaf spot ( Phaeoisariopsis griseola ) in two bean ( Phaseolus vulgaris ) cultivars (Rosinha G-2 and Carioca), pre-infected or not with bean line pattern mosaic virus (BLPMV), were determined. Trials were conducted at temperatures in the range from 9 to 27°C for rust and from 12 to 30°C for angular leaf spot. Regardless of viral pre-infection, the effect of temperature on the four monocyclic components followed an optimum curve and could be described by a generalized beta function. Generally, angular leaf spot was favoured by higher temperatures with an optimum for disease severity between 24.2 and 28.3°C compared with 15.9–18.5°C for rust. Pre-infection with BLPMV did not change the shape of the optimum curves for all components, but significantly reduced lesion density and disease severity on both cultivars. The development rates during incubation and latent periods for both fungal diseases were not affected by BLPMV. Pre-infection with virus did not alter the ranking of cultivars with respect to resistance to both fungal diseases.