Biochemical and catalytic properties of three recombinant alcohol acyltransferases of melon. sulfur-containing ester formation, regulatory role of CoA-SH in activity, and sequence elements conferring substrate preference

Alcohol acyltransferases (AAT) play a key role in the biosynthesis of ester aroma volatiles in fruit. Three ripening-specific recombinant AATs of cantaloupe Charentais melon fruit (Cm-AAT1, Cm-AAT3, and Cm-AAT4) are capable of synthesizing thioether esters with Cm-AAT1 being by far the most active. All proteins, as well as AAT(s) extracted from melon fruit, are active as tetramers of around 200 kDa. Kinetic analysis demonstrated that CoA-SH, a product of the reaction, is an activator at low concentrations and an inhibitor at higher concentrations. This was confirmed by the addition of phosphotransacetylase at various concentrations, capable of modulating the level of CoA-SH in the reaction medium. Site-directed mutagenesis of some amino acids that were specific to the Cm-AAT sequences into amino acids that were consensus to other characterized AATs greatly affected the selectivity of the original protein and the number of esters produced.     

Blood supply and arteriography of the pelvic limb of the Southern caracara (Caracara plancus) and great egret (Ardea alba)

The aim of this study was to compare the arterial vascularization of the pelvic limb between southern caracara (Caracara plancus) and great egret (Ardea alba) by dissection and radiographic examinations. Five specimens of caracaras (three males and two females), and seven great egrets (five males and two females) were used. Barium sulphate and latex suspension were injected into the left ventricle of the birds. The radiographs were taken with the pelvic limbs in the ventrodorsal, dorsoplantar, mediolateral and lateromedial recumbency. Thereafter, the material was fixed in a 10% solution of formaldehyde and dissected. The pelvic limb received its arterial supply from two main vessels, the ischiatic and external iliac arteries. The ischiatic artery presented to be the principal artery of pelvic limb in the caracara and great egret. Several branches arised from the ischiatic and external iliac arteries were described. No gender differences were observed in both species. The caracara and great egret showed arteries similar to those reported for the ostrich and domestic fowl. According to the results of this study, it is suggested that the caracara has a pelvic limb with more arterial branches and larger arterial diameter than the great egret, which is probably related to the specific behaviour of these birds, since the caracara is a bird that exercise more their pelvic limbs to capture its prey when compared with the great egret.     

Antioxidant and biocidal activities of Carum nigrum (seed) essential oil, oleoresin, and their selected components

In the present study, chemical constituents of the essential oil and oleoresin of the seed from Carum nigrum obtained by hydrodistillation and Soxhlet extraction using acetone, respectively, have been studied by GC and GC-MS techniques. The major component was dillapiole (29.9%) followed by germacrene B (21.4%), beta-caryophyllene (7.8%), beta-selinene (7.1%), and nothoapiole (5.8%) along with many other components in minor amounts. Seventeen components were identified in the oleoresin (Table 2) with dillapiole as a major component (30.7%). It also contains thymol (19.1%), nothoapiole (15.2.3%), and gamma-elemene (8.0%). The antioxidant activity of both the essential oil and oleoresin was evaluated in mustard oil by monitoring peroxide, thiobarbituric acid, and total carbonyl and p-anisidine values of the oil substrate. The results showed that both the essential oil and oleoresin were able to reduce the oxidation rate of the mustard oil in the accelerated condition at 60 degrees C in comparison with synthetic antioxidants such as butylated hydroxyanisole and butylated hydroxytoluene at 0.02%. In addition, individual antioxidant assays such as linoleic acid assay, DPPH scavenging activity, reducing power, hydroxyl radical scavenging, and chelating effects have been used. The C. nigrum seed essential oil exhibited complete inhibition against Bacillus cereus and Pseudomonas aeruginosa at 2000 and 3000 ppm, respectively, by agar well diffusion method. Antifungal activity was determined against a panel of foodborne fungi such as Aspergillus niger, Penicillium purpurogenum, Penicillium madriti, Acrophialophora fusispora, Penicillium viridicatum, and Aspergillus flavus. The fruit essential oil showed 100% mycelial zone inhibition against P. purpurogenum and A. fusispora at 3000 ppm in the poison food method. Hence, both oil and oleoresin could be used as an additive in food and pharmaceutical preparations after screening.     

Predictive equations of maintenance energy requirement for healthy and chronically ill adult dogs

Maintenance energy requirement (MER) is the energy amount necessary for dogs to maintain their weight and body condition. Some factors can influence the MER, such as gender, age, neutering status and also diseases. The present retrospective study aimed to evaluate MER of adult dogs with several diseases and compare with the MER of healthy adult dogs, observing the influence of parameters such as body condition score (BCS), neutering status, gender, age, diagnosis and type of food on MER of these dogs. A total of 165 adult dogs with weight changes of ≤5% were included and divided in groups according to diagnosis. Mean MER for healthy dogs was 86.09 kcal/BW^0.75, which differed from NRC and FEDIAF recommendations for inactive adult dogs (p = .047). Lowest MERs were of the endocrinopathies (78.52 ± 19.32 kcal/BW^0.75), orthopaedic diseases (59.71 ± 19.30 kcal/BW^0.75) and neurologic diseases (78.83 ± 32.66 kcal/BW^0.75) groups. Gastrointestinal diseases (99.59 ± 20.36 kcal/BW^0.75), orthopaedic diseases (59.71 ± 19.30 kcal/BW^0.75) and neoplasia (95.61 ± 21.02 kcal/BW^0.75) groups were the only groups that differed from the mean MER of healthy adult dogs. Regarding BCS, for each increasing point in a 9-point scale, there was a decrease of 9.8 kcal/BW^0.75 on MER, independent of diagnosis (p < .0001; r² = .55). There was no difference regarding breed size gender and age, but neutered dogs presented lower MER (p = .031). Based on data obtained from the present study, it can be concluded that it is necessary to consider BCS, age, neutering status and diagnosis when calculating MER, both in healthy dogs and chronically ill dogs.     

Cellular and humoral immune responses during intrathoracic paracoccidioidomycosis in BALB/c mice

Paracoccidioidomycosis is a chronic infection that primarily affects the lungs. Here we investigated cellular and humoral immune responses after intrathoracic Paracoccidioides brasiliensis infection in BALB/c mice. P. brasiliensis-colony-forming units (CFUs), fungal DNA and granulomas in lungs increased progressively, peaking at day 90 postinfection (p.i.). IFN-γ production was highest on day 15 p.i., declining thereafter. The kinetics of the NO production was similar to that described for IFN-γ. In contrast, IL-10 increased from day 45 p.i. reaching a peak at day 90. Levels of serum IgG1 were higher than IgG2a between days 30 and 90 p.i. 30% of mice died by day 90 p.i. These data indicate that infection with P. brasiliensis by the intrathoracic route shows high IFN-γ and NO production at day 15 p.i., unable to control multiplication of fungi, which appears to be associated with a progressive increase in IL-10 and in the number and complexity of granulomas.     

Effects of restricted vs. step up dietary adaptation for 6 or 9 days on feedlot performance,feeding behaviour,ruminal and blood variables of Nellore cattle

This study was designed to determine effects of quantitatively restricted intake of the final finishing diet as a means of dietary adaptation compared with diets increasing in concentrate (step up) over periods of 6 and 9 days on feedlot performance, carcass characteristics, feeding behaviour, blood gas profile and rumen morphometrics of Nellore cattle. One hundred twenty 22‐month‐old Nellore bulls (352 ± 20 kg) were randomly allocated in 24 pens and fed for 84 days. The experimental design was a completely randomized block with 2 × 2 factorial arrangement of treatments: adaptation using either step up protocol for 6 days or 9 days or restriction protocol for 6 days or 9 days, where the pen (five bullocks/pen; six pens/treatment) was considered the experimental unit. Cattle adapted by step up protocol had significantly greater average daily gain throughout the study when compared to those adapted by restriction protocol, regardless of adaptation length. During the adaptation period, a greater dry matter intake fluctuation was observed for animals maintained on restriction programme when compared to those adapted to the finishing diets using step up protocol. In addition, a protocol × length interaction for rumenitis scores was observed, where the level of injuries to the ruminal wall was significantly higher for the cattle adapted by restriction protocol for 9 days and lower for animals submitted for 9 days to the step up programme. Likewise, a protocol × length interaction for mitotic index was observed, in which cattle adapted by step up protocol for 9 days had lower numbers of cells in mitosis. Thus, the step up protocol led to better feedlot performance, and based on this fact, the length of 9 days promoted better rumen epithelium adaptation of Nellore cattle to high‐concentrate diets.     

Seasonal dynamics of microbial community in an aquaculture system for Nile tilapia (Oreochromis niloticus)

Studies on bacterial abundance and diversity will improve the understanding of the microbial ecology to optimize aquaculture production, water quality, disease control and environmental impact of effluents. We comparatively evaluated an aquaculture system in dry and rainy seasons by Fluorescent in situ Hybridization (FISH) and culture‐dependent methodology. Overall, a negative correlation between seasonality in bacterial and flagellates abundance was observed. Alpha‐, beta‐, gamma‐proteobacteria and Cytophaga‐Flavobacter were observed with seasonal variation. Putative pathogenic bacteria such as coagulase‐negative Staphylococcus, non‐fermenter Gram‐negative rods (Pseudomonas sp. and Burkholderia sp.), and Enterobacteriaceae were also detected in this study with significant seasonal variation. Focusing on medically important bacteria, our data show that microbial diversity in the environment associated with aquaculture, as it is practiced, may be altered in growth ponds used for fish cultivation. As an ecological consequence, potentially pathogenic bacteria might be released in high concentrations to the downstream environments posing potential threats to human and animal health.     

NH3 Volatilization from Urea-NBPT in Eucalyptus

High losses of nitrogen (N) by volatilization of ammonia from urea applied in Eucalyptus are expected due to the influence of plant residues on the soil surface. The study evaluated the N losses by volatilization of ammonia from urea coated with Thiophosphate N-(n-butil) triamide (NBPT) applied in soil with eucalyptus residues in surface under moisture treatments: fertilization in dry soil without irrigation; fertilization in dry soil with posterior irrigation depth (3 mm); fertilization in moist soil without irrigation and fertilization in moist soil with irrigation depth (3 mm). NBPT is a potential inhibitor of urease. Urea with NBPT shows lower losses by volatilization of ammonia when it is applied in dry soil; however in soil conditions of high moisture the losses as well as inhibitor effect of the NBPT are lower. The inhibitor effect of NBPT is reduced over time when it is subjected to moisture conditions.     

Influence of sexually inactive bucks subjected to long photoperiod or testosterone on the induction of estrus in anovulatory goats

The objective of this study was to evaluate the efficacy of treating sexually inactive bucks with artificial long photoperiod or testosterone on the induction of estrus in anovulatory grazing goats. A total of 91 multiparous mixed-breed anestrous goats were randomly assigned to one of three treatment groups: (1) joining with bucks subjected to 2.5 month of artificial long days (16 h of light/day; n = 31), (2) joining with testosterone-treated bucks (n = 30), and (3) joining with untreated bucks (control; n = 30). There were no differences between the light-treated (100%) and testosterone-treated (93%) bucks in their ability to induce estrus in anovulatory does. On the other hand, none of the goats in contact with control bucks exhibited estrus. The interval from start of mating to estrus was shorter in goats with the light-treated bucks (37.9 ± 4.8 h) compared with does in contact with testosterone-treated bucks (58.3 ± 8.7 h). The overall pregnancy rate in goats joined with light-treated, testosterone-treated and control bucks was 84%, 77% and 0%, respectively, with no difference (P > 0.05) between the first two groups. Anogenital sniffing, approaches, mounting attempts, and mounts were highest (P < 0.01) in light-treated bucks and lowest in control bucks. It was concluded that testosterone-treated bucks and long-day-treated bucks were equally effective in synchronizing estrus in anovulatory goats and resulted in similar levels of fertility. Given that light-treated bucks are unviable in communal production systems of goats raised by resource-poor farmers, the sexual arousal of bucks with testosterone is a practical and reliable method to induce ovulation in anovulatory goats in pastoral goat systems in hot environments.     

Effect of dietary high-oleic-acid oils that are rich in antioxidants on microsomal lipid peroxidation in rats

In contrast to other metabolic functions, the role of dietary antioxidants and oil on microsomal lipid oxidation has been less extensively studied. This study examines ascorbate-Fe(2+) and NADPH-induced lipid peroxidation of hepatic microsomes of rats that were fed for three weeks high-oleic-acid oils (high-oleic sunflower oil, HOSO; olive oil, OO; or olive pomace oil, OPO) containing different concentrations of the antioxidants alpha-tocopherol, erythrodiol, and oleanolic acid. The fatty acid composition of hepatic microsomes of Wistar rats that were fed for three weeks with the above-mentioned oils had lower proportions of C16:0, C18:2n6, and C22:6n3 and higher proportions of C18:0 and C18:1n9 than rats fed the control diet. Light emission by hepatic microsomes increased, in the first 180 min, 2-fold after ascorbate-Fe(2+) addition compared with NADPH addition. Both increases were less pronounced in microsomes of OPO-fed rats and to a smaller extent in microsomes of OO-fed rats. Smaller increases in light emission were associated positively with higher concentrations of dietary alpha-tocopherol, erythrodiol, and oleanolic acid but were not associated with changes in fatty acid composition of hepatic microsomes. Addition of alpha-tocopherol, erythrodiol, or oleanolic acid decreased light emission of hepatic microsomes with a greater inhibition in microsomes of rats fed the control diet. Our data suggest that erythrodiol and oleanolic acids partly explain the protective effect of dietary OPO on microsomal lipid peroxidation in rats.