Evaluation of serum- and animal protein-free media for the production of infectious bronchitis virus (M41) strain in a continuous cell line

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Some aspects of chicken behavior and welfare

Brazil is the world leader in broiler production and export. It achieved this position mainly to its excellent supply chain structure and climate, which favor poultry and grain production throughout its territory. Although Brazilian egg production is not as important as broiler production, this segment presents great potential of increasing its share in the global market. However, as elsewhere in the world, Brazilian poultry production faces the challenge to balance two elements within its supply chain: cruelty and productivity. The consumers of the European Union (EU) are very concerned with animal welfare issues. In order to increase its share in the European market, and eventually in the world market, Brazilian poultry producers must understand the effects of production systems on poultry welfare, and try to develop systems that are suited for its climate and other production conditions. There is a consensus that the natural behaviors performed by poultry in intensive production systems allow better welfare. This objective of this review is to present scientific research studies that relate different behaviors to chicken welfare. Poultry behavior is a reflex of their welfare status at a particular moment, and it is related to internal (physiological) and external (environmental) factors. Several natural behaviors that favor welfare, as well as undesirable behaviors, may be stimulated by environmental enrichment. The correct interpretation of the behaviors expressed by poultry, including their frequency, duration, and sequence, may be used to estimate their welfare. Animal production is an import sector of Brazilian economy. It significantly contributes to the Gross Domestic Product (GDP), in terms of products destined both to domestic consumption and exports. New technologies applied to products and management practices have been developed for field application, aiming at improving producers' productivity and profitability. In order to comply with the European Union's (EU) guidelines for animal protein production, Brazilian poultry production needs to undergo a process of adaptation. In May, 2007, the EU Commission established its new guidelines for animal welfare in poultry production, pressured by consumer demand. In the EU, there is a growing concern among consumers as to how poultry are reared and slaughtered. European consumers are in the fore front of the demand of high quality products produced with under better welfare conditions, and have spread this concern throughout the world. Beaumont et al. (2010) mentioned that European consumers frequently perceive that standard commercial poultry production has poor animal welfare practices. According to Nääs et al. (2008), Brazilian poultry production today needs to find a balance between cruelty and productivity. In fact, ensuring animal welfare may provide better financial results, as it increases the producer's profit margins and allows maintaining Brazilian chicken export quotas to the EU. França (2008) noted that biological studies that define ethical limits and guidelines for poultry production foster the development of new production practices that may ensure good product quality and productivity without putting bird welfare at risk. Gonyou (1994) states that, when animal welfare started to be studied, the only behavioral factors considered were those related to feeding and reproduction. These first studies used as indicators of animal welfare reduced life expectancy, impaired growth, impaired reproduction, body damage, disease, immunosuppression, adrenal activity, behavior anomalies, and self-narcotization (Broom, 1991). However, current studies evaluate additional indicators, such as natural behaviors, behavioral needs, preferences, behavioral problems, emotional state, cognitive abilities, etc. In the field of ethology, the expression of natural behavior is a frequently used tool used to estimate the welfare of poultry destined to human consumption. According to Bracke & Hopster (2006), natural behavior can be defined as the behavior the animal normaly presents when exposed to conditions similar to its natural habitat. Natural behaviors are pleasurable and promote biological functions that are meaningful to the animal's welfare. The definition of natural behavior, though, does not include the bird's behavior when sick, in flight or during aggression, since these are not considered pleasurable situations. Considering layer behavioral needs in the design of housing facilities optimize their welfare. Mishra et al. (2005) verified that ISA Brown layers spent, during 24 hours, around 97% of the time in the nest, feeding, walking, resting, or dust bathing, and that 57% of these behaviors did not depend on environmental enrichment. It was also observed that hens had preferred behavioral sequences, which included foraging and comfort behaviors, such as wing-stretching and preening. The present review aims at discussing, albeit not exhaustively, scientific research studies on the behavior of Gallusgallus domesticus and its relationship to welfare. The following behaviors are reviewed: feather pecking, scratching, dust bathing, nesting, locomotion activities, and aggressive behaviors. These behaviors are the most frequently observed in commercial broiler, broiler breeder, and layer farming, and therefore, monitoring their incidence may contribute to measure poultry welfare.

Poultry offal meal in broiler chicken feed

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparison of lipase production on crambe oil and meal by Fusarium sp (Gibberella fujikuroi complex)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

MINERAL NITROGEN SUBSTITUTION BY CHICKEN LITTER IN SUCCESSION OAT/CORN

The constant search for sustainability of production systems have driven research to find alternatives to the problems arising from the intensified use such systems. In this context the present work aimed study the effects of substitution of mineral nitrogen by chicken litter in oat and corn crop in succession and the chemical characteristics of soil. The study was conducted during the period May 2009 to March 2010 in area of Oxisol. The design was of randomized block with four replications. The six treatments were obtained by a combination of different amounts of chicken litter (0, 1500, 3000, 4500, 6000 and 7500 kg ha(-1)) applied 30 days before the sowing of oats combined with the mineral nitrogen applied in coverage in corn (311.1, 257.8, 202.2, 148.9, 95.6, 42.2 kg ha(-1) of urea), for the total supply of 140 kg ha(-1) of nitrogen (N). The application of poultry litter in oat promotes increased the production of dry matter, and content and accumulation of N. The mineral nitrogen substitution by chicken litter increases the yield of corn crop. The use of poultry litter alters the chemical properties of soil, increasing the levels of organic matter, exchangeable Al and acidity potential. However lowers the pH, K, Ca, Mg, sum of bases and base saturation.

Chicken meat consumer profile in northwestern São Paulo State, Brazil

Brazil is the world’s first chicken meat exporter nowadays. The maintenance of this position requires a constant quality attributes evolution. This work evaluated the chicken meat consumer profile in the northwest region of São Paulo state, the most important Brazilian poultry meat consumer market, in order to provide information to the productive sector. The data were collected using 482 interviews and questionnaires that were answered by e-mail. The questionnaires involved questions related to the consumer identification, habits and preferences and their knowledge about food safety, production system, sustainability and animal welfare. Most of the consumers, 62%, were female, with ages ranging from 20 to 50 years. Beef was preferred by the majority of the answerers and chicken and pork meat were together the second choice. Only 2% of the interviewed consumers mentioned not enjoying poultry meat. The main part of consumers, 67%, prefer to buy breast and leg cuts and only 11% are used to buy the whole poultry carcass. More than 60% of the interviewed have already eaten free range chicken meat, but the majority of them, 89%, are used to consume regular industrialized poultry. About 75% of the consumers believe hormones are used to grow the birds. Over 80% of people observe the expiration date before buying the product, but only 55% check if it has the stamp of the official inspection service. Color and appearance of meat are the most important factors that influence the consumer’s choice. The amount of water that drips on the tray is a rejection factor to 88% of answerers. Most of them, 66%, prefer lighter colored meat. Only 27% of them believe that chicken meat causes an environmental impact and 48% do not know the meaning of animal welfare. More than half of the interviewed do not consider animal welfare aspects before consuming any kind of meat. From these results obtained, it is possible to conclude that any effort to improve the product quality, mainly concerned to animal welfare and sustainability aspects, requires prior educational initiatives.

Evaluation of the use of probiotic acid lactic bacteria in the development of chicken hambúrguer

The objective of this study was to investigate the effect of fermentation with Lactobacillus acidophilus CRL 1014 on the physicochemical, microbiological and sensory characteristics of a hamburger product like processed with chicken meat and okara flour, with reduction of curing salts. A mixture of ingredients containing 90% chicken meat and 10% okara flour was subjected to the following treatments: F1: fermented with Lactobacillus acidophilus; F2:75 mg nitrite/kg and fermented with Lactobacillus acidophilus; F3: 150 mg nitrite/kg and unfermented. The quality of the “hamburgers” was assessed by physical and chemical analysis (pH, cooking yield and shrinkage), chemical composition, microbiological tests (Salmonella spp., count of sulphite-reducing clostridia, staphylococos coagulase-positive, total coliforms and Escherichia coli) and sensory analysis (sensory acceptance and purchase intent). During the first six days of fermentation, there was a decrease in pH from approximately 6.33 to 5.10. All the samples showed the same chemical composition (p < 0.05). The fermentation process was observed to inhibit the multiplication of microorganisms of several groups: coagulasepositive staphylococci, sulphite-reducing clostridia, Salmonella spp. and E. coli. The different “hamburgers” formulations showed high scores for all the sensory attributes evaluated, without differing from each other (p < 0.05). The results showed that the use of L. acidophilus CRL 1014 enabled the production of a safe product, with good physicochemical and sensory characteristics, in the absence of curing salts.

Emergence of enteric viruses in production chickens is a concern for avian health

Several viruses have been identified in recent years in the intestinal contents of chickens and turkeys with enteric problems, which have been observed in commercial farms worldwide, including Brazil. Molecular detection of these viruses in Brazil can transform to a big threat for poultry production due to risk for intestinal integrity. This disease is characterized by severely delayed growth, low uniformity, lethargy, watery diarrhea, delayed feed consumption, and a decreased conversion rate. Chicken astrovirus (CAstV), rotavirus, reovirus, chicken parvovirus (ChPV), fowl adenovirus of subgroup I (FAdV-1), and avian nephritis virus (ANV) were investigated using the conventional polymerase chain reaction (PCR) and the reverse transcription polymerase chain reaction (RT-PCR). In addition, the infectious bronchitis virus (IBV), which may play a role in enteric disease, was included. The viruses most frequently detected, either alone or in concomitance with other viruses, were IBV, ANV, rotavirus, and CAstV followed by parvovirus, reovirus, and adenovirus. This study demonstrates the diversity of viruses in Brazilian chicken flocks presenting enteric problems characterized by diarrhea, growth retard, loss weight, and mortality, which reflects the multicausal etiology of this disease

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.

Comparison of genotypes and antibiotic resistances of Campylobacter jejuni and Campylobacter coli on chicken retail meat and at slaughter

Multilocus sequence typing (MLST) and antibiotic resistance patterns of Campylobacter jejuni and Campylobacter coli from retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.

Production of avian leukosis virus particles in mammalian cells can be mediated by the interaction of the human immunodeficiency virus protein Rev and the Rev-responsive element.

In human immunodeficiency virus type 1-infected cells, the efficient expression of viral proteins from unspliced and singly spliced RNAs is dependent on two factors: the presence in the cell of the viral protein Rev and the presence in the viral RNA of the Rev-responsive element (RRE). We show here that the HIV-1 Rev/RRE system can increase the expression of avian leukosis virus (ALV) structural proteins in mammalian cells (D-17 canine osteosarcoma) and promote the release of mature ALV virions from these cells. In this system, the Rev/RRE interaction appears to facilitate the export of full-length unspliced ALV RNA from the nucleus to the cytoplasm, allowing increased production of the ALV structural proteins. Gag protein is produced in the cytoplasm of the ALV-transfected cells even in the absence of a Rev/RRE interaction. However, a functional Rev/RRE interaction increases the amount of Gag present intracellularly and, more strikingly, results in the release of mature ALV particles into the supernatant. RCAS virus containing an RRE is replication-competent in chicken embryo fibroblasts; however, we have been unable to determine whether the particles produced in D-17 cells are as infectious as the particles produced in chicken embryo fibroblasts.

Palm kernel meal in broiler diets: effect on chicken performance and health

An increase in the production of palm kernel meal (PKM) coupled with the concern for continued availability of conventional feedstuffs in some parts of the world has led to research to establish the maximum inclusion level of palm kernel meal in broiler diets. The results suggested that palm kernel meal has no anti-nutritional properties and thus its inclusion is safe up to at least 40% in the diet, provided the diet is balanced in amino acids and metabolisable energy. Although feed digestibility is decreased due to high dietary fibre when PKM is included in the diet, the feed intake is increased. This makes total digestible nutrient intake relatively high. beta-mannan is the main component of palm kernel meal non-starch polysaccharide (NSP). Both mannose and manno-oligosaccharides have been reported to act as prebiotics. The inclusion of palm kernel meal in the diet improves the immune system of birds and reduces pathogenic bacteria and increases the population of nonpathogenic bacteria in the intestine. These two benefits should be considered as strong recommendations for using palm kernel meal in broiler diets, particularly in palm kernel meal producing countries, not only for increasing bird productivity but also to improve chicken health. Selective enzyme addition increases feed efficiency and digestibility as well as decreasing the moisture content of faeces.

The chicken ovalbumin upstream promoter-transcription factors modulate genes and pathways involved in skeletal muscle cell metabolism

The chicken ovalbumin upstream promoter-transcription factors ( COUP-TFs) are orphan members of the nuclear hormone receptor ( NR) superfamily. COUP-TFs are involved in organogenesis and neurogenesis. However, their role in skeletal muscle ( and other major mass tissues) and metabolism remains obscure. Skeletal muscle accounts for similar to 40% of total body mass and energy expenditure. Moreover, this peripheral tissue is a primary site of glucose and fatty acid utilization. We utilize small interfering RNA ( siRNA)-mediated attenuation of Coup-TfI and II ( mRNA and protein) in a skeletal muscle cell culture model to understand the regulatory role of Coup-Tfs in this energy demanding tissue. This targeted NR repression resulted in the significant attenuation of genes that regulate lipid mobilization and utilization ( including Ppar alpha, Fabp3, and Cpt-1). This was coupled to reduced fatty acid beta-oxidation. Additionally we observed significant attenuation of Ucp1, a gene involved in energy expenditure. Concordantly, we observed a 5-fold increase in ATP levels in cells with siRNA-mediated repression of Coup-TfI and II. Furthermore, the expression of classical liver X receptor ( LXR) target genes involved in reverse cholesterol transport ( Abca1 and Abcg1) were both significantly repressed. Moreover, we observed that repression of the Coup-Tfs ablated the activation of Abca1, and Abcg1 mRNA expression by the selective LXR agonist, T0901317. In concordance, Coup-Tf-siRNA-transfected cells were refractory to Lxr-mediated reduction of total intracellular cholesterol levels in contrast to the negative control cells. In agreement Lxr-mediated activation of the Abca1 promoter in Coup-Tf-siRNA cells was attenuated. Collectively, these data suggest a pivotal role for Coup-Tfs in the regulation of lipid utilization/cholesterol homeostasis in skeletal muscle cells and the modulation of Lxr-dependent gene regulation.