Identification of phytotoxins from <Emphasis Type="Italic">Botryosphaeria obtusa</Emphasis>, a pathogen of black dead arm disease of grapevine

A bioassay-guided fractionation of a culture filtrate of Botryosphaeria obtusa led to the isolation of four dihydroisocoumarins, named mellein 1, 4-hydroxymellein 2, 7-hydroxymellein 3 and the new 4,7-dihydroxymellein 4. LC-UV-DAD-MS analysis of vine wood infected by B. obtusa revealed the presence of mellein (1). Botryosphaeria obtusa was also able to oxidise wood δ -resveratrol into the dimer delta-viniferin. The structures of isolated phytotoxins were established on the basis of IR, MS, 1D and 2D NMR.     

The Antinociceptive and Anti-Inflammatory Activities of Caulerpin,a Bisindole Alkaloid Isolated from Seaweeds of the Genus Caulerpa

The antinociceptive and anti-inflammatory activity of caulerpin was investigated. This bisindole alkaloid was isolated from the lipoid extract of Caulerpa racemosa and its structure was identified by spectroscopic methods, including IR and NMR techniques. The pharmacological assays used were the writhing and the hot plate tests, the formalin-induced pain, the capsaicin-induced ear edema and the carrageenan-induced peritonitis. Caulerpin was given orally at a concentration of 100 μmol/kg. In the abdominal constriction test caulerpin showed reduction in the acetic acid-induced nociception at 0.0945 μmol (0.0103–1.0984) and for dypirone it was 0.0426 μmol (0.0092–0.1972). In the hot plate test in vivo the inhibition of nociception by caulerpin (100 μmol/kg, p.o.) was also favorable. This result suggests that this compound exhibits a central activity, without changing the motor activity (seen in the rotarod test). Caulerpin (100 μmol/kg, p.o.) reduced the formalin effects in both phases by 35.4% and 45.6%, respectively. The possible anti-inflammatory activity observed in the second phase in the formalin test of caulerpin (100 μmol/kg, p.o.) was confirmed on the capsaicin-induced ear edema model, where an inhibition of 55.8% was presented. Indeed, it was also observed in the carrageenan-induced peritonitis that caulerpin (100 μmol/kg, p.o.) exhibited anti-inflammatory activity, reducing significantly the number of recruit cells by 48.3%. Pharmacological studies are continuing in order to characterize the mechanism(s) responsible for the antinociceptive and anti-inflammatory actions and also to identify other active principles present in Caulerpa racemosa.     

Intake,digestibility, performance,and carcass traits of beef cattle of different gender

The performance, intake, feed efficiency, and carcass traits of beef cattle from different gender profile were assessed. Fifteen animals (five steers, five spayed heifers, and five intact heifers) with ±250 kg of initial body weight were randomly assigned in individual pens and fed the same diet for 106 days. At the end of the trial, all the animals were slaughtered and the pH, temperature, and weight of the carcass were recorded. The right side of each carcass was then separated into chuck, shoulder, flank sirloin, and round for evaluation of commercial cuts yield. The left carcass sides were ribbed between the 12th and 13th ribs where the rib eye area and fat thickness measurements were taken. The 9th–11th rib section was removed from the left half carcass and then dissected into muscle, fat, and bones in order to estimate carcass composition. Gender had no effect (P > 0.05) on performance, intake, digestibility of dry matter and all the nutrients evaluated, feed efficiency, and carcass characteristics. It can be concluded that steers and heifers (spayed or not) have the same potential to produce beef. From a productive and welfare standpoint, there is no reason to spay heifers.     

Economical and financial analysis of lamb finishing fed with diets formulated according to the NRC (1985) and the NRC (2007)

This study compares both versions of the nutritional requirement system determined by the National Research Council (NRC) version 1985 (NRC85) and NRC version 2007 (NRC07), for finishing lambs in feedlots. Nineteen crossbred lambs were divided in four groups representing four experimental treatments: one diet according to NRC85 and three diets according to NRC07. The diets recommended by NRC07 considers crude protein intake relative to ruminal undegradable protein at 20, 40, and 60 % levels (NRC07/20, NRC07/40, and NRC07/60). Diets were composed of Brazilian semi-arid native grass silage, soybean meal, corn, annatto byproduct, and limestone. Purchases and sales of lambs were done according to average market prices in Brazil. The economic indicators considered pointed that all treatments were viable but NRC07/20 and NRC07/60 were more profitable with similar net present values (NPVs) and internal return rates (IRRs). NRC07/20 was the best option showing an IRR of 17.20 % and a payback period (PP) of 5.07 considering a fixed annual interest rate of 6 %. Sensitivity analysis considering a 10 % raise in variable costs showed negative NPVs, IRRs inferior to the opportunity cost rates adopted and PPs that exceeded the planning horizon of 7 years for both NRC85 and NRC07/40.     

Potential synergistic effect of Melia azedarach fruit extract and Beauveria bassiana in the control of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in cattle infestations

The use of a concentrate emulsion of Melia azedarach green fruits and a suspension of the fungus Beauveria bassiana was evaluated in the control of Rhipicephalus microplus on artificially infested cattle. The evaluation was conducted following the protocol established by the Brazilian Agriculture Ministry. Five groups of 4 or 5 animals were allocated to one of the following treatments: emulsion concentrate of M. azedarach at 0.25% (T AZED 0.25%), emulsion concentrate of M. azedarach at 0.5% (T AZED 0.5%), B. bassiana at 2.4 × 10(8) conidia (T BASS), association of the concentrate of M. azedarach at 0.25% with B. bassiana at 2.4 × 10(8) conidia (T AZED 0.25%+BASS), and control (untreated). The association of the two compounds provided better results than any one isolated treatment, indicating compatibility or perhaps a synergy between M. azedarach and B. bassiana. This treatment resulted in fewer engorged females (129 ± 70) than in the control group (233 ± 82), showing high performance against all developmental stages of the tick. Results revealed an apparent synergistic effect of M. azedarach and B. bassiana in the control of R. microplus that should be further investigated.     

Expression of the PEPT1, CAT,SOD2 and GPX1 genes in the zebrafish intestine supplemented with methionine dipeptide under predation risk

This study evaluated the effect of methionine supplementation, predation risk and their interaction on gut histology, whole-body cortisol levels, and intestinal gene expression in zebrafish. A total of 360 one-year-old animals were maintained under two environmental conditions and fed diets containing different methionine sources. Fish were fed either a control diet (CTL, without methionine supplementation), a diet supplemented with dl -methionine (DLM), or a diet supplemented with methionine dipeptide (MM) in the absence (AP) of a predator or in the presence of the predator (PP) for 48 h or 20 days. Predator-induced stress for 20 days resulted in lower body weight. Zebrafish fed methionine-supplemented diets had higher weight gain than control fish. We found no effect of predation stress or methionine supplementation on cortisol level. Predation risk and methionine supplementation showed no interaction effect on dipeptide transporter gene expression. After 48 h of predation pressure, zebrafish had higher mRNA expression of SOD2, CAT and GPX1 in the gut. After 20 days of exposure to the predator, zebrafish fed methionine-supplemented diets had lower expression of GPX1, SOD2 and CAT than those diet CTL. Methionine dipeptide and free methionine supplementation improved growth, intestinal health and survivability of zebrafish both conditions.     

Mineral salt intake effects on faecal-N concentration and the volume and composition of beef cattle urine

The effect of mineral salts on water ingestion and urine volume in cattle has been extensively studied. However, recently, this effect has been investigated as a potential mitigator of environmental aspects related to the nitrogen (N) cycle, such as nitrate (NO₃^?) lixiviation, ammonia (NH₃) volatilisation, and nitrous oxide (N₂O) emissions. The effect of mineral salts, particularly sodium chloride (NaCl), on urine-N concentration, urine volume, the proportion of N compounds in the urine, and faecal-N concentration has not yet been explored in field conditions with respect to environmental aspects of beef cattle production. The present study investigated the effect of dietary mineral salt rates on these parameters. A Latin square (5?×?5) experimental design was utilised with five concentrations of mineral salts in the diet: 0.0, 2.0, 4.0, 6.0, and 8.0 g based on dry matter (DM) ingestion (g/kg DM). The nitrogen concentration in the urine and urine volume increased linearly. The total N excreted (g/day) via urine did not vary with increasing mineral salt concentrations. When evaluated, the N compounds of urine (urea-N, allantoin-N, and hippuric acid-N) also reacted to the increased mineral salt concentrations, while creatinine-N did not. Urea-N, allantoin-N, and hippuric acid-N linearly increased their proportions in total N-urine. The N concentration in faeces was not affected by mineral salt concentrations. The urine volume, concentration of N, and proportion of N compounds in the urine affected N₂O emissions and NH₃ volatilisation. Therefore, mineral salt utilisation may be an option for mitigating N pollution from beef cattle, especially for grasslands in tropical countries.

     

Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.     

Total IgE and allergen-specific IgE and IgG antibody levels in sera of atopic dermatitis affected and non-affected Labrador- and Golden retrievers

Canine atopic dermatitis (CAD) is an allergic skin disease associated with IgE and IgG antibodies (Ab) to environmental allergens. The aim of this study was to determine which other factors influence serum Ab levels in CAD-affected and non-affected dogs as this has only been poorly investigated in dogs so far. Total and allergen-specific IgE levels and Dermatophagoides farinae (DF)-specific IgG1 and IgG4 were measured by ELISA in sera of 145 CAD-affected and 271 non-affected Labrador- and Golden retrievers. A multivariable logistic regression analysis including the factors age, breed, gender, castration, clinical CAD status and allergen-specific immunotherapy (ASIT) was performed. Golden retrievers had more frequently total (OR=1.87, 95% CI=1.26-2.87, p<0.01) and specific IgE levels above the threshold value than Labrador retrievers, suggesting that genetic factors influence IgE levels in dogs. Castration was generally associated with low Ab levels (OR=0.43-0.65, p<0.05). Surprisingly, dogs with CAD did not have increased odds for high IgE against any of the allergens tested. ASIT with DF was associated with high DF-specific IgG1 (OR=4.32, 95% CI 1.46-12.8, p<0.01) but was not associated with DF-specific IgG4 or decreased IgE levels. Further studies are needed to understand the role of allergen-specific IgE in CAD and of IgG1 in ASIT.