Mastoparan effects in skeletal muscle damage: An ultrastructural view until now concealed

Animal venoms have been valuable sources for development of new drugs and important tools to understand cellular functioning in health and disease. The venom of Polybia paulista, a neotropical social wasp belonging to the subfamily Polistinae, has been sampled by headspace solid phase microextraction and analyzed by gas chromatography-mass spectrometry. Recent study has shown that mastoparan, a major basic peptide isolated from the venom, reproduces the myotoxic effect of the whole venom. In this study, Polybia-MPII mastoparan was synthesized and studies using transmission electron microscopy were carried out in mice tibial anterior muscle to identify the subcellular targets of its myotoxic action. The effects were followed at 3 and 24 h, 3, 7, and 21 days after mastoparan (0.25 mu g/mu L) intramuscular injection. The peptide caused disruption of the sarcolemma and collapse of myofibril arrangement in myofibers. As a consequence, fibers presented heteromorphic amorphous masses of agglutinated myofilaments very often intermingled with denuded sarcoplasmic areas sometimes only surrounded by a persistent basal lamina. To a lesser extent, a number of fibers apparently did not present sarcolemma rupture but instead appeared with multiple small vacuoles. The results showed that sarcolemma, sarcoplasmic reticulum (SR), and mitochondria were the main targets for mastoparan. In addition, a number of fibers showed apoptotic-like nuclei suggesting that the peptide causes death both by necrosis and apoptosis. This study presents a hitherto unexplored view of the effects of mastoparan in skeletal muscle and contributes to discuss how the known pharmacology of the peptide is reflected in the sarcolemma, SR, mitochondria, and nucleus of muscle fibers, apparently its subcellular targets.

Animal welfare concepts and strategy for poultry production: a review

Well being of animals had been historically a public concern, since the beginning of human kind history. As world's population grows there is a need for food including meat. In the last decades there has been a great improvement in poultry production based on the careful control of several aspects, among which nutrition and management (environment, health and rearing systems). Nowadays, the search for good welfare conditions is a global tendency in animal production; however issues surrounding farm animal welfare or well-being, such as definitions, measurements, interpretation, and perception, continue to be controversial. It is known that the result of a broiler not adequately housed is a direct loss in production which leads towards a thought that health, welfare and productivity are intimately connected. In the other hand hints are found in the observation of behavioral responses as well as vocalization, which may provide more precise assessment to welfare. This has been possible due to the use of information technology applied to the field of ethology as well as the multidisciplinary view of the problem. This text provides a review on broiler's welfare issues since its definition to several way of trying to assess it adequately.

Importância do modelo animal para testar hipóteses sobre a fisiopatologia do binômio diabetes e incontinência urinária feminina

Aims: To discuss the importance of studying animal models to test hypotheses about the mechanisms of urinary continence and pathophysiology of diabetes and urinary incontinence. Source of Data: A literature review was conducted in PubMed and SciELO. The key words used were diabetes, urinary incontinence, urethra, human and rats. Summary of Findings: There is a strong relation between the genesis of urinary incontinence and diabetes mellitus. Due to the similarity of normal distribution of skeletal muscle and urethra anatomy between humans and rats, these animal models have been used in current research about these disorders. Conclusions: The use of rats as an animal model is suitable for experimental studies that test hypotheses about the mechanisms of continence and pathophysiology of the binomial diabetes mellitus and urinary incontinence, thus enabling solutions of great value in clinical practice.

Antioxidant activity and determination of total phenol contents, carotenoids, beta-carotene, lycopene and zinc in white, yellow and pinkish varieties of manihot esculenta crantz

There are cassava varieties that present compounds as carotenoids, beta-carotene, lycopene and minerals important for human and animal health. The present study evaluated the antioxidant activity of the white, yellow and pinkish varieties of Manihot esculenta, by mean of the DPPH test and by the ferrous ion-chelating activity. Furthermore, the total phenols, carotenoids, beta-carotene, lycopene and zinc contents were also determined. Utilizing the DPPH test it was possible to find that extracts of boiled samples presented higher antioxidant activity (89.53% -pinkish) in comparison to the fresh samples (1.97% -white). For the ferrous ion-chelating test, the highest activity was found for the boiled pinkish variety extract (63.43%) and the lowest was for fresh yellow extract (17.34%) the white sample did not present activity. The highest concentration of total phenols and zinc content was obtained for the boiled pinkish variety extract 136.12 mg EAG/g of extract and 0,811ppm, respectively, in the concentration of 1000 mu g/mL. The pinkish variety presented also higher quantity of pigments, including carotenoid (29.40 mu g/g), beta-carotene (9.14 mu g/100g) and lycopene (68.92%). According to the results obtained in this study it was possible to conclude that the yellow and pinkish varieties of M. esculenta present quantity of phenolic compounds and minerals sufficient to attribute the antioxidant activity and may thus contribute to reduce oxidative damage and be used as nutraceuticals or directly ingested in the diet to maintain good health.

Atividade antioxidante e determinação de fenóis totais, carotenoides, betacarotenos, licopeno e zinco em variedades branca, amarela e rosada de Manihot Esculenta Crantz

There are cassava varieties that present compounds as carotenoids, beta-carotene, lycopene and minerals important for human and animal health. The present study evaluated the antioxidant activity of the white, yellow and pinkish varieties of Manihot esculenta, by mean of the DPPH test and by the ferrous ion-chelating activity. Furthermore, the total phenols, carotenoids, beta-carotene, lycopene and zinc contents were also determined. Utilizing the DPPH test it was possible to find that extracts of boiled samples presented higher antioxidant activity (89.53% - pinkish) in comparison to the fresh samples (1.97% - white). For the ferrous ion-chelating test, the highest activity was found for the boiled pinkish variety extract (63.43%) and the lowest was for fresh yellow extract (17.34%) the white sample did not present activity. The highest concentration of total phenols and zinc content was obtained for the boiled pinkish variety extract 136.12 mg EAG/g of extract and 0,811ppm, respectively, in the concentration of 1000 µg/mL. The pinkish variety presented also higher quantity of pigments, including carotenoid (29.40 µg/g), beta-carotene (9.14 µg/100g) and lycopene (68.92%). According to the results obtained in this study it was possible to conclude that the yellow and pinkish varieties of M. esculenta present quantity of phenolic compounds and minerals sufficient to attribute the antioxidant activity and may thus contribute to reduce oxidative damage and be used as nutraceuticals or directly ingested in the diet to maintain good health.

Conscientizar para o bem-estar animal: posse responsável

The community awareness on responsible ownership associated with public policies is key to the promotion of animal welfare. Most cities have a large population of dogs and cats wandering causing serious health problems and abuse. This project aimed to guide and inform the population aimed at animal welfare with the simple and direct approach to issues related to responsible ownership, management, nature conservation and zoonoses. The methodology developed to approach the project in two modules: the I and II, along public and private Elementary and High Schools in Araçatuba. Module I (Mod.I) is basedon the use of puppet theater and musicals, considering the target audience children 3-10 years and the second module (Mod 2) consists of presentations of lectures by undergraduate and graduate course of Veterinary Medicine FMVA UNESP campus Araçatuba, being the target audience of High School teenagers. Data collection proceeded by questionnaires, with questions regarding the care and practices to pets. The results revealed that there is a lack of information about animal welfare practice.

De um subproduto industrial para um produto destinado à alimentação animal: produção de biomassa bacteriana

A grande diversidade das atividades industriais ocasiona durante o processo produtivo a geração de resíduos sólidos, líquidos e gasosos, que podem poluir e/ou contaminar o solo, a água e o ar. Embora existam regulamentos para o descarte desses resíduos, a inobservância às regras, a ineficiência dos tratamentos despoluentes e a inoperância de órgãos fiscalizadores permitem que, ainda hoje, sejam lançados ao ambiente grandes cargas de poluentes. Isso se aplica às indústrias de alimentos de origem animal. Felizmente, a visão dos empresários do setor vem mudando, na medida em que a imagem de uma empresa que conta com produtos e processos ambientalmente responsáveis representa parte das estratégias competitivas atuais. O objetivo deste trabalho foi demonstrar como um efluente industrial pode ser transformado em matéria prima para a obtenção de um produto com valor comercial, utilizando tecnologias reconhecidas pela indústria e a atividade microbiana. O efluente foi obtido em indústria de abate e processamento de tilápias, apresentado (valores médios) pH = 9,4, DQO = 1.127 mg/L, óleos e graxas = 1.166 mg/L, nitrogênio total = 813 mg/L, coliformes a 30–35o C = 1,0x105 NMP/mL, coliformes a 45o C = 0,41 NMP/mL, bolores e leveduras = 4,6x103 UFC/mL e, ocasionalmente, contendo Salmonella sp e Aeromonas sp. Os tratamentos físicos aplicados ao efluente incluíram gradeamento, filtração (50µm) e pasteurização (65o C/30 min). O cultivo de Rubrivivax gelatinosus foi realizado sob anaerobiose em reatores de vidro durante 7 dias, em temperatura ambiente (30±5o C) e 2.000±500 lux. A recuperação da biomassa foi feita por filtração tangencial (0,2 µm; 1,5 bar), centrifugação (3.400 g/30 min; 5o C) e liofilização (-40o C) e a pulverização foi realizada manualmente. A produção de massa celular atingiu 0,18 g/L, com produtividade de 0,0634 g/L.dia. O processo promoveu redução de 52% na DQO, 48% em óleos e graxas e 22% no nitrogênio total, gerando um resíduo com pH 7,9, livre de bactérias patogênicas e, portanto, apto ao descarte. O produto obtido apresentou cor vermelho escuro (L = 22,42; C = 14,22; h = 25,48), 4,55% de umidade, 57,39% de proteína, 11,08% de extrato etéreo, 4,05% de matéria mineral, 3,03 mg/g de oxicarotenóides, 20,27 NMP/g de coliformes a 30–35o C, <1,0 NMP/g de coliformes a 45o C, 1,2x103 UFC/g de bolores e leveduras e ausência de microrganismos patogênicos. Essas características apontam para um potencial pigmentante e nutricional positivo no produto, que pode encontrar aplicação na produção animal, com segurança microbiológica. Dessa forma, fecha-se um ciclo autosustentável que pode ser adotado na própria fonte geradora do resíduo industrial, permitindo a reversão de gastos com tratamento despoluente em receitas advindas da comercialização de um novo produto.

Resolving conflicts with Canada geese: an animal welfare perspective

The scope and significance of human conflicts with urban and suburban Canada goose populations has been growing rapidly since the mid 1980s. A lack of basic understanding about the biology and ecology of locally abundant goose populations has led, in part, to argument between opposing camps over the appropriate approaches and methodologies to resolve human-goose conflicts. Animal welfare interests have focused on the humaneness of roundup and slaughter programs, and advocated non-lethal approaches coupled with what they view as the more benign population control activity of egg addling. Some traditional wildlife managers have argued that non-lethal approaches have been tried and have failed, and that procedures such as addling do not work quickly or effectively. Differences have led to legal confrontations that absorb considerable energy and effort and may make cooperative involvement more difficult. This paper articulates some of the arguments that comprises the basis for the perspective of animal welfarists. It ends with a call for greater cooperation and involvement between all interests concerned with Canada geese.

ANIMAL POPULATION ECOLOGY AND CONTROL FUNDAMENTALS

Expensive, extensive and apparently lethal control measures have been applied against many species of pest vertebrates and invertebrates for decades. In spite of this, few pests have been annihilated, and in many cases the stated goals have become progressively more modest, so that now we speak of saving foliage or a crop, rather than extermination. It is of interest to examine the reasons why animals are so difficult to exterminate, because this matter, of course, has implications for the type of control policy we pursue in the future. Also, it has implications for the problem of evaluating comparatively various resource management strategies. There are many biological mechanisms which could, in principle, enhance the performance of an animal population after control measures have been applied against it. These are of four main types: genetic, physiological, populationa1, and environmental. We are all familiar with the fact that in applying a control measure, we are, from the pest's point of view, applying intense selection pressure in favor of those individuals that may be preadapted to withstand the type of control being used. The well-known book by Brown (1958) documents, for invertebrates, a tremendous number of such cases. Presumably, vertebrates can show the same responses. Not quite so familiar is the evidence that sub-lethal doses of a lethal chemical may have a physiologically stimulating effect on population performance of the few individuals that happen to survive (Kuenen, 1958). With further research, we may find that this phenomenon occurs throughout the animal kingdom. Still less widely recognized is the fact that pest control elicits a populational homeostatic mechanism, as well as genetic and physiological homeostatic mechanisms. Many ecologists, such as Odum and Allee (1950, Slobodkin (1955), Klomp (1962) and the present author (1961, 1963) have pointed out that the curve for generation survival, or the curve for trend index as a function of last generations density is of great importance in population dynamics.

Application of chromatographic and spectroscopic techniques in the evaluation of the lipid fraction of animal products

Lipolysis and oxidation of lipids in foods are the major biochemical and chemical processes that cause food quality deterioration, leading to the characteristic, unpalatable odour and flavour called rancidity. In addition to unpalatability, rancidity may give rise to toxic levels of certain compounds like aldehydes, hydroperoxides, epoxides and cholesterol oxidation products. In this PhD study chromatographic and spectroscopic techniques were employed to determine the degree of rancidity in different animal products and its relationship with technological parameters like feeding fat sources, packaging, processing and storage conditions. To achieve this goal capillary gas chromatography (CGC) was employed not only to determine the fatty acids profile but also, after solid phase extraction, the amount of free fatty acids (FFA), diglycerides (DG), sterols (cholesterol and phytosterols) and cholesterol oxidation products (COPs). To determine hydroperoxides, primary products of oxidation and quantify secondary products UV/VIS absorbance spectroscopy was applied. Most of the foods analysed in this study were meat products. In actual fact, lipid oxidation is a major deterioration reaction in meat and meat products and results in adverse changes in the colour, flavour and texture of meat. The development of rancidity has long recognized as a serious problem during meat handling, storage and processing. On a dairy product, a vegetal cream, a study of lipid fraction and development of rancidity during storage was carried out to evaluate its shelf-life and some nutritional features life saturated/unsaturated fatty acids ratio and phytosterols content. Then, according to the interest that has been growing around functional food in the last years, a new electrophoretic method was optimized and compared with HPLC to check the quality of a beehive product like royal jelly. This manuscript reports the main results obtained in the five activities briefly summarized as follows: 1) comparison between HPLC and a new electrophoretic method in the evaluation of authenticity of royal jelly; 2) study of the lipid fraction of a vegetal cream under different storage conditions; 3) study of lipid oxidation in minced beef during storage under a modified atmosphere packaging, before and after cooking; 4) evaluation of the influence of dietary fat and processing on the lipid fraction of chicken patties; 5) study of the lipid fraction of typical Italian and Spanish pork dry sausages and cured hams.

Stem Cells as a therapy for myocardial infarction in animal models

Advances in stem cell biology have challenged the notion that infarcted myocardium is irreparable. The pluripotent ability of stem cells to differentiate into specialized cell lines began to garner intense interest within cardiology when it was shown in animal models that intramyocardial injection of bone marrow stem cells (MSCs), or the mobilization of bone marrow stem cells with spontaneous homing to myocardium, could improve cardiac function and survival after induced myocardial infarction (MI) [1, 2]. Furthermore, the existence of stem cells in myocardium has been identified in animal heart [3, 4], and intense research is under way in an attempt to clarify their potential clinical application for patients with myocardial infarction. To date, in order to identify the best one, different kinds of stem cells have been studied; these have been derived from embryo or adult tissues (i.e. bone marrow, heart, peripheral blood etc.). Currently, three different biologic therapies for cardiovascular diseases are under investigation: cell therapy, gene therapy and the more recent “tissue-engineering” therapy . During my Ph.D. course, first I focalised my study on the isolation and characterization of Cardiac Stem Cells (CSCs) in wild-type and transgenic mice and for this purpose I attended, for more than one year, the Cardiovascular Research Institute of the New York Medical College, in Valhalla (NY, USA) under the direction of Doctor Piero Anversa. During this period I learnt different Immunohistochemical and Biomolecular techniques, useful for investigating the regenerative potential of stem cells. Then, during the next two years, I studied the new approach of cardiac regenerative medicine based on “tissue-engineering” in order to investigate a new strategy to regenerate the infracted myocardium. Tissue-engineering is a promising approach that makes possible the creation of new functional tissue to replace lost or failing tissue. This new discipline combines isolated functioning cells and biodegradable 3-dimensional (3D) polymeric scaffolds. The scaffold temporarily provides the biomechanical support for the cells until they produce their own extracellular matrix. Because tissue-engineering constructs contain living cells, they may have the potential for growth and cellular self-repair and remodeling. In the present study, I examined whether the tissue-engineering strategy within hyaluron-based scaffolds would result in the formation of alternative cardiac tissue that could replace the scar and improve cardiac function after MI in syngeneic heterotopic rat hearts. Rat hearts were explanted, subjected to left coronary descending artery occlusion, and then grafted into the abdomen (aorta-aorta anastomosis) of receiving syngeneic rat. After 2 weeks, a pouch of 3 mm2 was made in the thickness of the ventricular wall at the level of the post-infarction scar. The hyaluronic scaffold, previously engineered for 3 weeks with rat MSCs, was introduced into the pouch and the myocardial edges sutured with few stitches. Two weeks later we evaluated the cardiac function by M-Mode echocardiography and the myocardial morphology by microscope analysis. We chose bone marrow-derived mensenchymal stem cells (MSCs) because they have shown great signaling and regenerative properties when delivered to heart tissue following a myocardial infarction (MI). However, while the object of cell transplantation is to improve ventricular function, cardiac cell transplantation has had limited success because of poor graft viability and low cell retention, that’s why we decided to combine MSCs with a biopolimeric scaffold. At the end of the experiments we observed that the hyaluronan fibres had not been substantially degraded 2 weeks after heart-transplantation. Most MSCs had migrated to the surrounding infarcted area where they were especially found close to small-sized vessels. Scar tissue was moderated in the engrafted region and the thickness of the corresponding ventricular wall was comparable to that of the non-infarcted remote area. Also, the left ventricular shortening fraction, evaluated by M-Mode echocardiography, was found a little bit increased when compared to that measured just before construct transplantation. Therefore, this study suggests that post-infarction myocardial remodelling can be favourably affected by the grafting of MSCs delivered through a hyaluron-based scaffold

Quality and safety of oils and fats obtained as co-products or by-products of the food chain and destined to animal production

The aim of the first part of this thesis was to evaluate the effect of trans fatty acid- (TFA), contaminant, polycyclic aromatic hydrocarbon (PAH)- and oxidation productenriched diets on the content of TFA and conjugated linoleic acid (CLA) isomers in meat and liver of both poultry and rabbit. The enriched feedings were prepared with preselected fatty co-and by-products that contained low and high levels of TFA (low, palm fatty acid distillate; high, hydrogenated palm fatty acid distillate), environmental contaminants (dioxins and PCBs) (two different fish oils), PAH (olive oil acid oils and pomace olive oil from chemical refining, for low and high levels) and oxidation products (sunflower-olive oil blend before and after frying), so as to obtain single feedings with three enrichment degrees (high, medium and low) of the compound of interest. This experimental set-up is a part of a large, collaborative European project (http://www.ub.edu/feedfat/), where other chemical and health parameters are assessed. Lipids were extracted, methylated with diazomethane, then transmethylated with 2N KOH/methanol and analyzed by GC and silver-ion TLC-GC. TFA and CLA were determined in the fats, the feedings, meat and liver of both poultry and rabbit. In general, the level of TFA and CLA in meat and liver mainly varied according to those originally found in the feeding fats. It must be pointed out, though, that TFA and CLA accumulation was different for the two animal species, as well as for the two types of tissues. The TFA composition of meat and liver changes according to the composition of the oils added to the feeds with some differences between species. Chicken meat with skin shows higher TFA content (2.6–5.4 fold) than rabbit meat, except for the “PAH” trial. Chicken liver shows higher TFA content (1.2–2.1 fold) than rabbit liver, except for the “TRANS” and “PAH” trials. In both chicken and rabbit meats, the TFA content was higher for the “TRANS” trial, followed by the “DIOXIN” trial. Slight differences were found on the “OXIDATION” and “PAH” trends in both types of meats. In both chicken and rabbit livers, the TFA content was higher for the “TRANS” trial, followed by those of the “PAH”, “DIOXIN” and “OXIDATION” trials. This trend, however, was not identical to that of feeds, where the TFA content varied as follows: “TRANS” > “DIOXIN” >“PAH” > “OXIDATION”. In chicken and rabbit meat samples, C18:1 TFA were the most abundant, followed by C18:2 TFA and C18:3 TFA, except for the “DIOXIN” trial where C18:3 TFA > C18:2 TFA. In chicken and rabbit liver samples of the “TRANS” and “OXIDATION” trials, C18:1 TFA were the most abundant, followed by C18:2 TFA and C18:3 TFA, whereas C18:3 TFA > C18:2 in the “DIOXIN” trial. Slight differences were found on the “PAH” trend in livers from both species. The second part of the thesis dealt with the study of lipid oxidation in washed turkey muscle added with different antioxidants. The evaluation on the oxidative stability of muscle foods found that oxidation could be measured by headspace solid phase microestraction (SPME) of hexanal and propanal. To make this method effective, an antioxidant system was added to stored muscle to stop the oxidative processes. An increase in ionic strength of the sample was also implemented to increase the concentration of aldehydes in the headspace. This method was found to be more sensitive than the commonly used thiobarbituric acid reactive substances (TBARs) method. However, after antioxidants were added and oxidation was stopped, the concentration of aldehydes decreased. It was found that the decrease in aldehyde concentration was due to the binding of the aldehydes to muscle proteins, thus decreasing the volatility and making them less detectable.