Myrciaria dubia, an Amazonian fruit: population structure and its implications for germplasm conservation and genetic improvement.

Myrciaria dubia (camu-camu) is an Amazon tree that produces a tart fruit with high vitamin C content. It is probably the fruit with the highest vitamin C content among all Brazilian fruit crops and it can be used to supplement daily vitamin C dose. This property has attracted the attention of consumers and, consequently, encouraged fruit farmers to produce it. In order to identify and select potential accessions for commercial exploitation and breeding programs, M. dubia has received considerable research attention. The identification and characterization of genetic diversity, as well as identification of the population structure of accessions preserved in germplasm banks are fundamental for the success of any breeding program. The objective of this study was to evaluate the genetic variability of 10 M. dubia populations obtained from the shores of Reis Lake, located in the municipality of Caracaraí, Roraima, Brazil. Fourteen polymorphic inter simple sequence repeat (ISSR) markers were used to study the population genetic diversity, which resulted in 108 identified alleles. Among the 14 primers, GCV, UBC810, and UBC827 produced the highest number of alleles. The study illustrated the suitability and efficiency of ISSR markers to study the genetic diversity of M. dubia accessions. We also revealed the existence of high genetic variability among both accessions and populations that can be exploited in future breeding programs and conservation activities of this species.

Evaluation of antioxidant capacity of 13 plant extracts by three different methods: cluster analyses applied for selection of the natural extracts with higher antioxidant capacity to replace synthetic antioxidant in lamb burgers.

The aims of this study were: to evaluate the total equivalent antioxidant capacities (TEAC) and phenolic contents of 13 plants extracts; to select the most promising extracts regarding reducing activity using cluster analysis multivariate statistical technique; and to analyse evaluate sensory acceptance of lamb burgers produced with the most promising natural antioxidants replacing sodium erythorbate. Plant extracts were evaluated regarding TEAC by DPPH(•) and FRAP methods, and total phenolics contents by Folin-Ciocalteau assay. The TEAC values ranged from 0.50 to 9.06 g trolox/100 g dry weight (dw) and from 43.6 to 472.32 µmol trolox/g dw for DPPH(•) and FRAP methods, respectively, and the total phenolic contents from 5.98 to 74.01 mg GAE/g dw. Extracts from Origanum vulgare, Melissa officinalis, Origanum majorana L. and Rosmarinus officinalis were grouped as the ones with higher antioxidant capacities by cluster analysis. All burgers produced with each one of these four plant extracts or with sodium erythorbate showed no differences (P > 0.05) regarding consumers' sensory acceptance. In conclusion, it is possible to replace sodium erythorbate in lamb burgers by any of the four natural extracts selected without compromising sensory acceptance of this meat product.

Effects of addition of different sources and doses of sugars on in vitro digestibilities of dry matter, fibre and cell wall monosaccharides of corn silage in ruminants.

In ruminant diets, soluble sugar is an important factor in the digestive process. The objective of this study was to evaluate the effects of the source and dose of soluble sugars, under controlled pH conditions, on the in vitro digestibility of DM, fibre fractions (NDF and ADF) and cell wall neutral monosaccharides of corn silage. Silage was collected from several points in a silage mass from a bunker silo, oven-dried at 55°C and ground through a 1-mm screen. Sub-samples were combined with sugars to compose the treatments, in a 5 × 5 factorial arrangement, as a combination of five soluble sugar sources (glucose, fructose, arabinose, xylose and sucrose) and five sugar doses (0, 100, 200, 300 and 400 g/kg sugar in DM corn silage), respecting the following proportions of sugar : corn silage, 0 : 100, 10 : 90, 20 : 80, 30 : 70, 40 : 60 represented by the sugar doses, respectively. An in vitro test was performed to determine the true digestibility (D) of the chemical entities (DM, NDF and ADF) and cell wall monosaccharides (glucose = gluc, arabinose = arab and xylose = xyl). During the first 12 h of incubation, the pH was maintained above 6.0 by the addition of 2.5 N NaOH. The concentrations of neutral monosaccharides (arabinose, xylose and glucose) were determined by GLC. The soluble sugars decreased the digestibility of corn silage followed by pH reduction, especially at doses higher than 200 g/kg sugar. Overall, xylose, followed by sucrose, fructose and arabinose, had greater impacts on DM digestibility, whereas fibre digestibility was impaired by sucrose at all doses. Xylose and fructose had greater impacts on NDF digestibility at 300 and 400 g/kg sugar. Although xylose impaired the Dgluc in the cell wall in all doses. All doses of glucose improved the Dgluc and Dxyl in the cell wall.

Performance, carcass characteristics and meat quality of Nellore cattle supplemented with supranutritional doses of sodium selenite or selenium-enriched yeast.

The enrichment of meat with selenium is important to improve the intake of selenium by humans. The effects of supranutritional doses of sodium selenite or selenium-enriched yeast on performance, carcass characteristics and meat quality were evaluated using 63 Nellore cattle in a completely randomized design with two sources (sodium selenite and selenium-enriched yeast), three levels (0.3, 0.9 and 2.7 mg Se/kg DM) and control treatment (without addition of selenium). Final body weight (BW), average daily gain, dry matter intake and gain to feed ratio (G : F) at the end of 84 days of supplementation were not influenced by treatments (P>0.05). Values of pH, ribeye area, back fat thickness and marbling score were also not influenced by treatments ( P>0.05). Dressing percentage was greater (P=0.02) in Nellore cattle supplemented with organic Se (58.70%) compared to animals supplemented with inorganic Se (57.94%). Hot carcass weight increased ( P=0.05) with the increasing of Se levels in the diet. Colour, shear force (SF), cooking and drip loss remained unchanged ( P>0.05); however thiobarbituric acid reactive substances was 15.51% higher with inorganic Se compared with organic Se. The selenium concentration in the meat of animals receiving organic selenium was higher ( P<0.001) than that of animals receiving sodium selenite, at all levels (0.3; 0.9 and 2.7 mg/kg DM). The meat of animals receiving 2.7 mg of organic Se/kg of DM presented concentration of 372.7 µg Se/kg in the L.dorsi muscle, and the intake of 150 g of this meat by humans provides approximately 100% of the recommended Se intake (55 µg Se/day for adults). Therefore, the use of supranutritional doses of 2.7 mg Se/kg of DM, regardless of source, is a way of naturally producing selenium-enriched meat without compromising performance, carcass characteristics and quality of Nellore bovine meat.

Visceral leishmaniasis after renal transplantation: report of 4 cases in northeastern Brazil.

Visceral leishmaniasis (VL) is a well recognized opportunistic infection in immunosuppressed patients, which may cause febrile illness. We describe 4 renal transplant patients with VL in an endemic area in Brazil and their response to therapy. In 3 cases the diagnosis was confirmed by bone marrow aspirate that revealed the presence of Leishmania. In 1 case the bone marrow aspirate was inconclusive and the diagnosis was made through spleen biopsy that showed the presence of the parasite. VL needs to be considered as a cause of febrile illness in transplanted patients living in endemic areas.

Maize plants produce direct resistance elicited by Tetranychus urticae Koch (Acari: Tetranychidae).

Plants can be attacked by a wide variety of herbivores. Thus, developing protective mechanisms for resistance against these agents is an advantage for survival and reproduction. Over the course of evolution, many resistance mechanisms against herbivory have been developed by the plants. Induced direct and indirect resistance mechanisms can manifest in plants after herbivore attack. The two-spotted spider mite Tetranychus urticae is not a pest of maize crops (Zea mays), despite being reported infesting plants that may have resistances against this herbivore. We tested the hypothesis that maize plants would be able to induce direct resistance against T. urticae after, evaluating the effect of T. urticae infestation in maize plants on the development and reproduction of conspecifics. We tested induced direct resistance performing infestation and measuring biological parameters upon a second infestation. Maize plants, 40 days after sowing, were divided into two groups: 30 not infested by T. urticae (clean plants clean) and, 30 infested by the spider mite. Infestation of maize plants by T. urticae reduced the conspecific female adult survival. However, no change in the survival of immature or reproduction was observed. These results suggest the induction of induced direct resistances in maize by T. urticae. This is first report of direct resistance induction in Z. mays by the two-spotted spider mite T. urticae.

Evaluation of oxidative stability of lamb burger with Origanum vulgare extract.

The objective was to evaluate replacement of sodium erythorbate with a natural antioxidant (oregano extract) on physicochemical and sensory stability of lamb burgers, and determine the appropriate amount. Five treatments were prepared, including control (without antioxidant), sodium erythorbate, and three concentrations of oregano extract (13.32, 17.79 and 24.01mL/kg), based on antioxidant capacity determined using the Folin-Ciocalteu, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) methods, respectively. Burgers containing oregano extract, at the concentration determined by FRAP method, had higher oxidative stability, evidenced by an 80% reduction (P<0.001) in thiobarbituric acid reactive substances, effective inhibition of protein oxidation (P<0.01) and less colour loss during frozen storage. Oregano extract did not impair (P>0.05) consumers' sensory acceptance of the lamb burgers. Under the conditions tested, addition of 24mL/kg of oregano extract could be recommended as a natural antioxidant in lamb burgers.

Carryover of aflatoxins from feed to lambari fish (Astyanax altiparanae) tissues.

The aim of this study was to verify the carryover of aflatoxin B1 from feed to lambari fish. Aflatoxins (AF) were incorporated into feed, checking the levels by HPLC. Treatments were: Control, feed without toxin; A, feed + 10 µg AFB1 kg-1; B, feed + 20 µg AFB1 kg-1; and C, feed + 50 µg AFB1 kg-1. Juveniles of lambari fish were placed in 12 aquariums at a density of 50 fish/m2. Fish were fed twice a day with extruded feed, at 5% of animal biomass. The unit sample was constituted by a pool of 10 fish. AFs B1, B2, G1, G2 and M1 were quantified by HPLC in fish muscle and liver after 30, 60, 90 and 120 days of experiment. There was accumulation of AFs is fish liver and muscle, mainly after 90 days. Fish from treatment C had higher levels of AFB1 in muscle when compared with the others, and AFB1 in muscle at 120 days was similar to the levels in feed. Therefore, when lambari fish is exposed on a daily and long-term basis to AFs in feed, the regulation limits for AFs in animal feed do not guarantee safety for consumers.

Phenolic acid protects of renal damage induced by ochratoxin A in a 28-days-oral treatment in rats.

The present study aimed to characterize the chlorogenic acid (ChlA) capacity to reverse the toxic effects induced by ochratoxin A (OTA) in a subacute toxicity test in rats. Male Wistar rats were fed orally by gavage for 28 days with OTA (0.4mg/kg bw/day), ChlA (5mg/kg bw/day) or the combination OTA (0.4mg/kg bw/day)+ChlA (5mg/kg bw/day). No deaths, no decrease in feed intake or body weight in any experimental group were recorded. The negative control group and the animals treated with ChlA alone showed no changes in any parameters evaluated. In OTA-treated group significant changes such as decrease in urine volume, proteinuria, occult blood, increase in serum creatinine values; decrease in absolute and relative kidney weight and characteristics histopathological lesions that indicated kidney damage were observed. However, limited effect on oxidative stress parameters were detected in kidneys of OTA-treated group. Animals treated with the combination OTA+ChlA were showed as negative control group in the evaluation of several parameters of toxicity. In conclusion, ChlA, at given concentration, improved biochemical parameters altered in urine and serum and pathological damages in kidneys induced by OTA exposure, showing a good protective activity, but not by an apparent antioxidant mechanism.

Turbidimetric determination of some sulphur compounds in ethanol from fermentation.

A simple method is described which can be used for the determination of certain sulphur compounds found in industrial ethanol obtained from fermentation of molasses. The method is based on the turbidimetric determination of sulphate after the sample has been treated with dilute hydrogen peroxide solution, by precipitation of the sulphate with barium chloride under appropriate conditions. Several samples of fermentation ethanol have been analysed by this method and the sulphur contents found compared with the total acidity.

Utilization of O-phthalaldehyde-sulphuric acid as a spray reagent in thin-layer chromatographic detection of some indolealkylamines and application to cutaneous secretion extracts of toad species.

An expansion of the utilisation of o-phthalaldehyde in sulphuric acid medium as spray reagent was carried out when tryptophan and some tryptophan-derived indole alkylamines such as tryptamine, serotonin, bufotenine, dehydrobufotenine and bufotenidine were examined by thin-layer chromatography. Rf-values and limits of detection ranging from 20 (serotonin) to 100 (dehydrobufotenine) ng per spot were found. Application of this reagent for the detection of some of these compounds was carried out, using either methanolic extracts or column chromatographic fractions of the skin secretion of the toads Bufo ictericus and Odontophrynus cultripes.

Cissampelos sympodialis Eichl (Menispermaceae) leaf extract induces interleukin-10-dependent inhibition of Trypanosoma cruzi killing by macrophages.

The aqueous fraction of the ethanolic extract (AFL) of Cissampelos sympodialis Eichl (Menispermaceae), popularly known as milona, has been shown to have both immunosuppressive and anti-inflammatory effects. In the present study we investigated the modulation of macrophage antimicrobicidal activity by in vitro treatment with the extract from C. sympodialis. Normal and thioglycolate-elicited mouse peritoneal macrophages were infected in vitro with the protozoan Trypanosoma cruzi DM28c clone. We observed that the AFL (used at doses ranging from 13 to 100 microg/ml) increased T. cruzi growth and induced a 75% reduction in nitric oxide production. This inhibition could be mediated by the stimulation of macrophage interleukin-10 (IL-10) secretion since the in vitro treatment with the AFL stimulated IL-10 production by T. cruzi-infected macrophages. These results suggest that the anti-inflammatory effect of the AFL from C. sympodialis could be, at least in part, mediated by the inhibition of macrophage functions and that the inhibition of macrophage microbicidal activity induced by the C. sympodialis extract may be mediated by the decrease in macrophage function mediated by interleukin-10 production.

Cervical spondylomyelopathy in Great Danes: a magnetic resonance imaging morphometric study.

Morphometric investigations comparing normal and affected animals increase our understanding of spinal diseases in dogs. The aim of this study was to generate morphometric data for osseous-associated cervical spondylomyelopathy (CSM) in Great Danes (GDs). Magnetic resonance imaging (MRI) morphometric features of the cervical vertebral column of GDs with and without clinical signs of CSM were characterized and compared. Thirty client-owned GDs were prospectively enrolled, including 15 clinically normal and 15 CSM-affected GDs. All dogs underwent MRI of the cervical to thoracic vertebral column (C2-C3 through T1-T2). Areas of the cranial and caudal articular processes, and the height, width and areas of the vertebral canal and spinal cord were determined. Middle foraminal heights were measured. Intervertebral disc width was measured before and after traction. Intraobserver and interobserver agreement were calculated. CSM-affected GDs had larger areas of the caudal articular processes from C2-C3 through T1-T2. In CSM-affected GDs, the vertebral canal and spinal cord areas were significantly smaller at C5-C6 and C6-C7, the vertebral canal width was significantly narrower at C6-C7 and C7-T1, and the spinal cord width was significantly narrower at C5-C6 and C6-C7. Middle foraminal height was smaller in CSM-affected GDs from C3-C4 through C7-T1. Neutral intervertebral disc widths were smaller in CSM-affected GDs. It was concluded that the cervical vertebral canal dimensions are significantly different between normal and CSM-affected GDs. Absolute vertebral canal stenosis and severe foraminal stenosis involving the cervical vertebrae distinguish CSM-affected from clinically normal GDs. These findings are relevant to the pathogenesis of osseous-associated CSM and should be taken into consideration when performing imaging studies and planning surgery.

Irradiated vacuum-packed lamb meat stored under refrigeration: microbiology, physicochemical stability and sensory acceptance.

Reducing spoilage and indicator bacteria is important for microbiological stability in meat and meat products. The objective was to evaluate the effect of different doses of gamma radiation on the shelf-life of lamb meat, vacuum-packed and stored under refrigeration, by assessing the microbiological safety, physicochemical stability and sensory quality. Lamb loin cuts (Longissimus dorsi) were irradiated with 1.5kGy and 3.0kGy. The samples, including control, were stored at 1±1°C during 56days. Samples were analyzed on zero, 14, 28, 42 and 56days by their microbiological and physicochemical characteristics. Sensory quality was carried out on day zero. The results showed a reduction (p<0.05) in the microbial load of the irradiated samples. The acceptance of lamb loins was not affected (p>0.05) by the radiation doses. Thus gamma irradiation at 3.0kGy was effective in reducing the content of microorganisms, without harming the physicochemical characteristics evaluated.

Maize plants produce direct resistance elicited by Tetranychus urticae Koch (Acari: Tetranychidae) / Plantas de milho produz resistência elicitada por Tetranychus urticae Koch (Acari: Tetranychidae)

Abstract Plants can be attacked by a wide variety of herbivores. Thus, developing protective mechanisms for resistance against these agents is an advantage for survival and reproduction. Over the course of evolution, many resistance mechanisms against herbivory have been developed by the plants. Induced direct and indirect resistance mechanisms can manifest in plants after herbivore attack. The two-spotted spider mite Tetranychus urticae is not a pest of maize crops (Zea mays), despite being reported infesting plants that may have resistances against this herbivore. We tested the hypothesis that maize plants would be able to induce direct resistance against T. urticae after, evaluating the effect of T. urticae infestation in maize plants on the development and reproduction of conspecifics. We tested induced direct resistance performing infestation and measuring biological parameters upon a second infestation. Maize plants, 40 days after sowing, were divided into two groups: 30 not infested by T. urticae (clean plants clean) and, 30 infested by the spider mite. Infestation of maize plants by T. urticae reduced the conspecific female adult survival. However, no change in the survival of immature or reproduction was observed. These results suggest the induction of induced direct resistances in maize by T. urticae. This is first report of direct resistance induction in Z. mays by the two-spotted spider mite T. urticae.

Resumo As plantas podem ser atacadas por uma grande diversidade de herbívoros. Sendo assim, o desenvolvimento de mecanismos de proteção para se defenderem destes agentes é um diferencial para a sobrevivência e reprodução. Ao longo da evolução, diversos mecanismos de resistência contra herbívoria foram desenvolvidos pelas plantas. Mecanismos de resistência induzida direta e indireta podem manifestar-se em plantas após o ataque de herbívoros. O ácaro-rajado Tetranychus urticae não é uma praga na cultura da cultura do milho (Zea mays), apesar de ser relatado infestando plantas, as quais podem apresentar resistências contra tal herbívoro. Testamos a hipótese de que plantas de milho seriam capazes de induzir resistências diretas à T. urticae após o ataque, avaliando o efeito da infestação de T. urticae de plantas em milho sobre seu desenvolvimento e reprodução de coespecíficos. Testamos resistência induzida direta realizando uma infestação e medindo os parâmetros biológicos em uma segunda infestação. Plantas de milho foram divididas em dois grupos: 30 plantas não por T. urticae (plantas limpas) e 30 plantas infestadas pelo ácaro-rajado. A infestação de plantas de milho por T. urticae reduziu a sobrevivência de fêmeas adultas do coespecífico. Entretanto, não foi observada alteração na sobrevivência das formas imaturas ou reprodução. Tais resultados sugerem a indução de resistências induzida diretas em milho por T. urticae. Este é primeiro relato de indução de resistência direta em Z. mays pelo ácaro-rajado T. urticae.

Breast skin-line detection using dynamic programming.

In this paper, we present a novel method to extract the breast skin-line based on dynamic programming. Skin-line extraction is an important preprocessing step in CAD systems; however, it is a challenging problem due to the presence of noise, underexposed regions, which results in a low contrast area near the skin-air interface, and artifacts such as labels. Our proposal utilizes the stroma edge to constrain searching for the border. In order to cope with noise, we consider several candidate points for the border interface which are obtained by the Laplace operator applied in pre-defined directions in the mammogram. The breast contour is obtained from the candidate points using a dynamic programming algorithm. This utilizes a criterion of optimality to obtain the optimum contour by minimization of a cost function. The method was evaluated using 82 mammograms whose contour were manually extracted by a radiologist from the mini-MIAS database. The Polyline Distance Measure was evaluated for each contour selected with the proposed method, obtaining a mean error of 2.05 pixels and a standard deviation of 0.80.

Momento de aplicação do nitrogênio e algumas variáveis estruturais e bromatológicas do capim-massai / Time of nitrogen application and some structural and bromatologic variables of massaigrass

As plantas forrageiras tropicais apresentam alto potencial de produção de massa de forragem, condicionada às condições edafoclimáticas, principalmente à adubação nitrogenada. A magnitude da resposta depende do sincronismo entre a disponibilidade de nutrientes e as necessidades da planta forrageira na rebrotação, e os resultados encontrados na literatura são contrastantes. Assim, este trabalho foi realizado com o objetivo de avaliar características estruturais e bromatológicas de Panicum maximum Jacq cv. Massai submetido a quatro doses de nitrogênio (0, 40, 80 e 120mg/dm3) e três momentos de aplicação após o corte (um, três e sete dias), divididos em três cortes, em um esquema fatorial 4 x 3 e delineado inteiramente ao acaso, com quatro repetições. O experimento foi conduzido em casa de vegetação. O incremento na taxa de aparecimento de perfilhos, na densidade populacional de perfilhos, na produção de massa seca de folha total e nos teores de proteína bruta foi de 42%, 65%, 770% e 35%, respectivamente, para a dose de 120mg/dm³ em relação à ausência de adubação nitrogenada. As fibras em detergente neutro e em detergente ácido decresceram cerca de 6% e 9%, respectivamente, com a adição de nitrogênio. Apenas a massa seca de folha total respondeu às épocas de aplicação de nitrogênio após o corte, com valores pouco expressivos. A adubação nitrogenada exerce efeito positivo sobre as variáveis estudadas, porém novos estudos devem ser conduzidos para a definição da época de adubação de nitrogênio após o corte, para a melhor utilização do nutriente pelas plantas.(AU)

Tropical forage plants have a high potential for mass production of fodder, subject to soil and climatic conditions, especially with nitrogen fertilization. The magnitude of the response depends on the timing of availability of nutrients and forage plant needs in the regrowth. Thus, this work was carried out to evaluate structural and bromatologic characteristics of the Panicum maximum Jacq cv. Massai in response to four nitrogen doses (0, 40, 80 and 120mg/dm3) and with three application times after cutting (one, three and seven days) in a 4 x 3 factorial treatment combination, in a completely randomized design, with four replicates. The experiment was conducted in a greenhouse. The increase in the rate of tillering, tiller population density, dry mass total leaf production and crude protein levels was 42%, 65%, 770% and 35%, respectively for the dose of 120mg / dm³ regarding the absence of fertilization. The neutral detergent and acid detergent fibers showed a decrease of 6% and 9% with nitrogen addition. Dry mass total leaf production responded to times of nitrogen application after cutting, with little significant values. Nitrogen fertilization has a positive effect on the variables studied, but new studies should be conducted to define the timing of nitrogen fertilization after cutting, for better utilization of this nutrient by plants.(AU)

In vitro evaluation of Clostridium septicum alpha toxoid / Avaliação in vitro de toxoide alfa de Clostridium septicum

Aiming to investigate in vitro alternatives, a test for neutralizing antibody detection using cell culture was developed. This test was more sensitive than previous animal models, allowing for detection of substantially lower alpha toxin and anti-alpha toxin titers. Titers observed during in vivo and in vitro seroneutralization had a correlation of 99.12 percent, indicating that cell culture is a viable alternative in the evaluation of vaccine potency, screening of vaccinal seeds, and Clostridium septicum alpha toxin titration.

Padronizou-se um teste para detecção de anticorpos neutralizantes in vitro, em cultura de células. O modelo in vitro mostrou-se mais sensível que os testes com animais, permitindo a detecção de títulos de toxina e antitoxina alfa mais baixos. Os títulos observados na soroneutralização in vivo e in vitro, apresentaram correlação de 99,12 por cento, demonstrando ser a cultura de células uma alternativa viável na avaliação da potência de vacinas, triagem de sementes vacinais e titulação de toxina alfa de Clostridium septicum.