Direct HPLC analysis of quercetin and trans-resveratrol in red wine,grape, and winemaking byproducts

A simple and fast reversed-phase HPLC method using diode array detection was developed and validated for the simultaneous determination of trans-resveratrol and quercetin in Sicilian red wine from the Nero d'Avola red grape variety. Investigation was also extended to the quantitative determination of resveratrol and quercetin in grape skins and winemaking byproducts obtained from the same cultivar. Samples were eluted using a C18 narrow-bore column under isocratic conditions in less than 20 min. Quantification of trans-resveratrol and quercetin in red wine was performed without any sample pretreatment, whereas the determination of these phenolic compounds in grape skins and wine pomage required a solvent extraction procedure. Linearity was demonstrated over the 0.39-12.5 and 0.45-57.6 microg/mL range for trans-resveratrol and quercetin, respectively. Detection limits in real samples were in the low ppm level (0.07 mg/L for trans-resveratrol and 0.12 mg/L for quercetin). The HPLC-UV/DAD method was applied for the routine analyses of red wine and grape skin and winemaking byproduct extracts to evaluate their trans-resveratrol and quercetin content. In particular, a very high content of quercetin was found in wine pomace, suggesting the use of this wine byproduct as a potential source of this health-promoting phenolic compound.     

Aeta Magbukún of Mariveles: Traditional Indigenous Forest Resource Use Practices and the Sustainable Economic Development Challenge in Remote Philippine Regions

The Aeta Magbukún of Mariveles are one of the least known and researched Indigenous peoples remaining on the fringe of the bay, and within the remaining forests in Bataan province on Luzon Island in the Philippines. This work describes the unique cultural systems and language of the Aeta Magbukún tribe in Biaan, Mariveles, and both the traditional forest resource use and the evolving new subsistence practices developed to adapt to the encroachment of non-Indigenous peoples onto ancestral lands. The Aeta's forest resource use practices are discussed from a sustainable Indigenous development context within unique socioeconomic, cultural, and environmental circumstances in Bataan.     

A Case of Bronchioloalveolar Carcinoma in a Mare

A case of pulmonary carcinoma in a 23-year-old Quarterhorse mare is described. On physical examination, depression, poor body condition, tachypnea, bilateral serosanguineous nasal discharge and a wide area of reduced breath sounds in association with the right hemithorax were detected. Laboratory evaluation showed leukocytosis with neutrophilia and lymphopenia, hyperfibrinogenemia, hyperprotidemia, hypoalbuminemia, increased beta 2 and gamma globulin fractions, hypoxemia, and normocapnia. Radiography and thoracic ultrasonography revealed a large rounded mass extending from the 9th to the 14th right intercostal space, where neither bronchial nor vascular structures were detectable. Endoscopy showed a large amount of serosanguineous fluid within the tracheal lumen and a mass of reddish soft tissue completely obliterating the right caudal lobar bronchus. Histopathology of the endobronchial mass and of the transthoracic ultrasound-guided biopsy samples was consistent with a primary pulmonary epithelial tumor. Due to worsening of the clinical condition, the mare was euthanized. Postmortem examination confirmed the presence of a large 30-cm mass located in the right caudal pulmonary lobe, characterized by epithelial cells arranged in papillary projections and alveolar structures, findings consistent with bronchioloalveolar carcinoma.     

Malignant peripheral nerve sheath tumor as a cause of chronic cardiac insufficiency in cattle

Chronic cardiac insufficiency was associated with a malignant peripheral nerve sheath tumor in a cow. An eight-year-old cow developed a progressive condition (over a period of three months) characterized by an enhanced abdominal volume, reluctance to move, a positive jugular pulse, watery diarrhea and death. At necropsy, moderate subcutaneous edema and an enhanced hepatic lobular pattern were observed. A 23x20x11 cm firm, grayish-white mass adhered to and infiltrated the right atrium. Multiple firm, yellowish-white nodules of 0.5 to 12 cm in diameter were diffusely scattered in the epicardium and parietal pericardium. Histologically, the tumor was poorly circumscribed with foci of infiltration of the myocardium. The neoplastic cells had two major histologic patterns, Antoni types A and B. Within occasional foci, pleomorphic cells with an epithelioid appearance were present in addition to multinucleated cells with periodic acid Schiff (PAS)-positive cytoplasmic globules. Foci of cartilaginous and granular differentiations were interspersed among the neoplastic cells. Multiple vessels presented wall hyalinization and tumoral embolus. Large necrotic foci with mineralization and cholesterol clefts were also observed. Immunohistochemically, the tumor was positive for S100 protein, vimentin and neuron-specific enolase labeling.     

Simulation of over-exploitation of ornamental fish and its consequences for the functional structure of assemblages of Amazonian streams

The consequences of over-exploitation may seriously impair the integrity and functioning of ecosystems. When loss of species is accompanied by the loss of ecological processes, the effects are no longer just taxonomic and may affect the stability of the environments. Ornamental fishing is one of the main economic activities of the middle Negro River. Such activity is directed to a small number of species and has the Cardinal Tetra (Paracheirodon axelrodi) as the main target species. Given the potential effects this pressure and the lack of information on its consequences, the present study analysed, through simulations, the possible effects of depletion of ornamental fish populations on the functional structure of fish assemblages, represented by functional diversity (FD) and functional redundancy (FR) indexes. We sampled 13 streams exploited by the ornamental fishery, where we collected 4,286 specimens of 110 species, of which 22 were targeted as ornamental fishing. We found that the ornamental species corresponded, on average, to 27.8% of the species and 33.3% of the FD of the assemblages. In a scenario of complete exclusion of ornamental species, local communities would lose 24.6% of the FD and reduce up to 12% of the FR. With these results, it is possible to infer that the local extinction of exploited species would cause negative impacts on the multifunctionality and the resilience of the streams ecosystems. In this way, we emphasise the need for proper management of the ornamental fisheries aiming to the sustainability of the activity and to the conservation of ecosystem functionality.     

Reproducible infection model for Clostridium perfringens in broiler chickens

Experiments were carried out to establish an infection and disease model for Clostridium perfringens in broiler chickens. Previous experiments had failed to induce disease and only a transient colonization with challenge strains had been obtained. In the present study, two series of experiments were conducted, each involving four groups of chickens with each group kept in separate isolators. A coccidial vaccine given at 10 times the prescribed dosage was used to promote the development of necrotic enteritis. In the first experiment, cultures of C. perfringens were mixed with the feed at day 9, 10, 11, and 12, and the coccidial vaccine was given at day 10, whereas in the second experiment, C. perfringens cultures were mixed with the feed at day 17, 18, 19, and 20, and the coccidial vaccine was given at day 18. Chickens were examined at day 9, 11, 12, and 15 (Experiment 1), and at day 17, 18, 20, and 24 (Experiment 2). There was no mortality in any of the groups; however, chickens in the groups receiving both coccidial vaccine and C. peifringens developed the subclinical form of necrotic enteritis, demonstrated by focal necroses in the small intestine, whereas chickens in control groups or groups receiving only coccidial vaccine or only C. perfringens cultures developed no necroses. The results underline the importance of predisposing factors in the development of necrotic enteritis.     

Supplementation of L‐glycine and L‐glutamate to Japanese quails from 01 to 36 days of age using the ideal protein concept

Experiment I: T_1‐1 = basal diet with 25% crude protein (CP) + limiting amino acids (LA); T_1‐2 = 20% CP + LA; T_1‐3 = 20% CP + LA + L‐glycine; T_1‐4 = 20% CP + LA + L‐glutamate; T_1‐5 = 20% CP + LA + L‐glycine + L‐glutamate. Experiment II: T_2‐1 = basal diet with 22% CP + LA; T_2‐2 = 20% CP + LA; T_2‐3 = 17.6% CP + LA + L‐glycine; T_2‐4 = 17.6% CP + LA + L‐glutamate; T_2‐5 = 17.6% CP + LA + L‐glycine + L‐glutamate. The reduction of dietary protein based on the concept of ideal protein decreases nitrogen excretion in quails when L‐glycine is added to the diets. Quails fed diets supplemented with L‐glutamate as the non‐specific nitrogen source equivalent to the nitrogen level of the control diet had increased nitrogen excretion. However, quails had reduced nitrogen excretion in both experiments when L‐glycine was added to diets with L‐glutamate. Carcass fat was increased by reducing dietary protein, but fat deposition was reduced by adding L‐glutamate and L‐glycine, or both. The dietary addition of L‐glutamate and L‐glycine in quails based on the ideal protein concept is not necessary (Exp. I). Although the total nitrogen, electrolytic balance and glycine level were adjusted in diets, quails had decreased performance. Therefore, other hypotheses besides protein reduction need to be studied (Exp. II). Protein reduction with supplementation of only limiting essential amino acids does not affect quail performance. Dietary addition of L‐glycine reduces nitrogen excretion.     

A novel strategy for resynchronization of ovulation in Nelore cows using injectable progesterone (P4) and P4 releasing devices to perform two timed inseminations within 22 days

We aimed to evaluate the reproductive performance of Nelore lactating cows submitted to a resynchronization 12 days after timed artificial insemination (TAI) with or without a long‐acting progesterone (P4‐LA) treatment. Nelore cows were submitted to a P4/oestradiol‐based TAI protocol (D0 = insemination). On D12, cows in the control group (n = 184) received a new P4 intravaginal device (0.96 g), whereas cows in the P4‐LA group (n = 192) received the P4 device and 75 mg P4‐LA. Cows identified as non‐pregnant (n = 120) by regression of corpus luteum using colour Doppler ultrasonography on D20 had the P4 device removed and received 500ug of sodium cloprostenol, 1 mg of oestradiol cypionate and 300 IU of eCG and were re‐inseminated on D22. There was no difference (p > 0.10) in the pregnancy rate at D20, D30 and D60 after first TAI between the control (69%, 59.7% and 57%, respectively) and P4‐LA (67%, 55.7%, and 55.2%, respectively) groups. Pregnancy losses were similar between both groups (p > 0.1). For cows submitted to the second TAI, the pre‐ovulatory follicle size did not differ (p > 0.1), but the oestrous detection and pregnancy rates were greater (p < 0.05) in the P4‐LA group (92.2% [59/64] and 60.9% [39/64], respectively) than in controls (75% [42/56] and 44.6% [25/56]). The cumulative pregnancy rate after two TAIs did not differ (p > 0.1) between control (73.3% [135/184]) and P4‐LA (76% [146/192]) groups. The use of P4‐LA at 12 days after TAI potentially increases the pregnancy rates for a new early resynchronization strategy associated with the Doppler imaging for pregnancy diagnosis and results in an alternative to perform two TAIs in 22 days in beef cows.     

Sperm cryopreservation of tiete tetra Brycon insignis (Characiformes): effects of cryoprotectants,extenders, thawing temperatures and activating agents on motility features

The aim of this study was to test the effects of cryoprotectants [dimethyl sulphoxide (DMSO) and methylglycol], extenders (0.9% NaCl, 5% glucose, Beltsville Thawing Solution^? and Merck III^?), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) on the cryopreservation process of tiete tetra Brycon insignis sperm. Sperm was loaded in 0.5 mL straws, frozen in nitrogen vapour at ?170 °C and stored in liquid nitrogen. Post‐thaw sperm quality was evaluated in terms of subjective motility rate, quality motility score (0=no movement; 5=rapidly swimming spermatozoa), duration of motility and vitality (eosin–nigrosin staining). Post‐thaw sperm motility rate was greater in methylglycol (76–88%), compared with DMSO (23–59%). In general, the highest quality motility scores were observed when sperm was thawed at 30 °C and triggered in 1% NaHCO₃ (3.5–4.3). Duration of motility was longer when triggered in 1% NaHCO₃ (95–120 s) compared with 0.29% NaCl (69–107 s). Sperm vitality was not affected by any of the parameters tested and varied from 51% to 69% intact sperm. Brycon insignis sperm frozen in methylglycol combined with any of the extenders tested and using the methods described above yields motility above 57% and that should last long enough to fertilize oocytes.     

Effect of live food transition time on survival,growth and metamorphosis of yellowtail clownfish, <Emphasis Type="Italic">Amphiprion clarkii</Emphasis>, larvae

Amphiprion clarkii, a popular marine ornamental fish, despite having an already established reproduction technology, still presents divergences regarding the feeding protocol used for its larviculture in what concerns the ideal day of transition from rotifers to Artemia. This study aimed to determine the optimum time to start supplying Artemia to larvae. Survival, growth, weight gain and metamorphosis of treatments: (T2) start of Artemia supply at the 2nd day after hatching (DAH); (T4) start at the 4th DAH; and (T6) start at the 6th DAH were evaluated. Survival rates ranged from 60 to 66 %. Fish in T2 had a number of metamorphosed fish statistically higher than T6 and began to metamorphose 1 day sooner than other treatments. The positive results obtained for T2 can be related to the premature ability of this species to capture large live food, which can provide many benefits to the larvae. It is concluded that it is possible to offer Artemia nauplii to yellowtail clownfish at 2 DAH.