Detection of chromosomal bla(CTX-M-2) in diverse Escherichia coli isolates from healthy broiler chickens

The rise of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in food-producing animals is a growing concern for public health. We investigated ESBL producers isolated from broiler chickens in Brazil and characterized 19 CTX-M-2-producing E.coli. The ISCR1 was detected upstream of the chromosome-located gene bla(CTX-M-2), associated with sul-1 type integron structure. CTX-M-2-producing E.coli exhibited different PFGE-types and phylogenetic groups, showing a non-clonal dissemination. The sequence types found (ST93, ST155 and ST2309) have been associated with humans and animals worldwide. Herein, we report the chromosomal location of bla(CTX-M-2) on E.coli, highlighting the risks of multidrug-resistant bacteria in food-producing animals.

Digestive enzymatic responses of chickens feed-restricted and refed as affected by age

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Seroprevalence of salmonella and mycoplasma in commercial broilers, backyard chickens, and spent hens in the region of Triangulo Mineiro, state of Minas Gerais, Brazil

Avian salmonellosis and mycoplasmosis are infectious diseases that, in addition of causing lack of flock uniformity, represent a hazard to human health. The objective of the present study was to evaluate the seroprevalence of mycoplasmosis and salmonellosis in commercial broilers, backyard chickens, and spent hens slaughtered at a processing plant with local health inspection in Uberlandia, MG, Brazil. A total of 210 samples were randomly collected at the time of bleeding. Samples were submitted to rapid plate serum agglutination test (RSA) for the classification of Salmonella pullorum, Salmonella gallinarum, Mycoplasma gallisepticum and Mycoplasma synoviae. In order to increase result specificity, mycoplasmosis-positive samples were submitted to hemagglutination inhibition test (HI). No samples presented detectable antibodies against Salmonella pullorum or Salmonella gallinarum in the RSA test. Only Mycoplasma synoviae was detected in 14% of the backyard chickens and 0.74% in commercial broilers, whereas no antibodies were detected in spent hens. The seroprevalence rates found in the present study emphasize the need of keeping chicken flocks free from disease using effective biosafety systems.

Intestinal integrity and performance of broiler chickens fed a probiotic, a prebiotic, or an organic acid

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

Genetic and phenotypic parameters of carcass and organ traits of broiler chickens

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic

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

Strategic use of dietary electrolyte balance and ratio for broiler chickens

The literature reports dealing with the dietary electrolyte ratio (K+Cl)/ Na are rare, although the concept has been proposed by Mongin in 1981. Thus, its application appears to be limited as a nutritional strategy in feed formulation, which usually meets only the minimum nutritional recommendations in Na, K and Cl. The objective of this study was to evaluate the performance of broilers submitted to different dietary electrolyte balances (DEB) Na+K-Cl and dietary electrolyte ratio (DER) from 1 to 21 d of age. A total of 1575 male 1-d old broiler chicks were randomly assigned to 5 treatments with 9 replicates of 35 chicks each. The treatments consisted of diets with 5 relation of electrolytes, arising from combinations DEB/DER 150/3, 250/2, 250/3, 250/4 and 350/3, with use of NaCl, NaHCO3, KCl, K2SO4 and CaCl2. All diets were corn-soybean meal based and formulated to meet or exceed the NRC (1994) requirements. Chicks had ad libitum access to feed and water in floor pens with wood shavings as litter. Body weight, feed intake and feed conversion ratio were measured at 21 d of age. It was found that only the feed conversion was significantly affected (P = 0.0142) by the combinations of relation of electrolytes (DEB and DER). The supplementation levels in DEB and DER were made to fit the data into a surface analysis to allow increasing levels of DEB (150–350 mEq/kg) and narrow and broad ratio of DER (2–4). For the canonical analysis of response surface was obtained the stationary point for body weight (DEB = 255.77 mEq/kg, and DER = 2.73:1) the value was 942.02 g; for feed intake (DEB = 251.69 mEq / kg and DER = 3.51:1), the value was 1200.02 g; and for feed conversion (DEB = 254.62 mEq/kg and DER = 3.06:1) the value was 1.35. The results of this experiment indicated that the best performance was obtained with combinations of relation of electrolytes for DEB between 251 and 255 mEq/kg and DER between 2.73:1 and 3.5:1.

Stunning method in broiler chickens meat processing

Quality is a variable concept, which involves many factors, depending on the consumer market. In meat production, the concern with environmental aspects, animal welfare and the health and safety of workers is increasing. This work studied the effect of controlled atmosphere stunning of broilers on meat features and biochemical parameters for stress. Cobb broilers were stunned by electrical stunning and by controlled atmosphere with 70% CO2 and 70% CO2 + 30% Argon. After stunning, serum levels of glucose, lactate and corticosterone were compared with those of broilers at rest, immediately before transportation and slaughter and after 12 h of feed withdrawal (control group). At slaughter, blood volume drained during bleeding was not different for the stunning methods tested, ranging from 3.3 to 3.4% birds weight. This finding was important to demonstrate that gas stunning was not responsible for the animals′ death. Final pH in breast (6.1 to 6.2) and thigh (6.3 to 6.5) also did not vary among the different stunning methods (P > 0.05). Lightness (L = 60.55) and redness (a = +8.94) values found for breasts from electrical stunning showed that they were darker and redder (P < 0.05), probably due to changes in blood pressure. Glucose and corticosterone levels were not different between gas stunned birds (302.45 to 315.7 mg/dl and 55.71 to 72.49 ng/ml respectively) and birds at rest (305.95 mg/dl and 50.65 ng/ml) (P > 0.05). These stress indicators were higher (337.65 mg/dl for glucose and 104.13 ng/ml for costicosterone) when electrical stunning was used (P < 0.05). Lactate concentrations were lower (5.4 mmol/l) for birds at rest (P < 0.05) but not different for all stunning methods tested (7.3 to 8.1 mmol/l; P > 0.05). These results show that serum glucose may be used as a stress indicator in birds, with the advantage of being a quick and cheap biochemical test. Gas stunning favored birds′ management during slaughter and so reduced workers′ effort and injury hazard and the amount of feces and dust in the room. To make this method available for a large scale process, adjustments in equipments will be necessary to avoid delays in the processing line.

Accuracy of nonlinear feed formulation for broiler chickens: maximising broiler profit

The modeling technique is simple, useful and practical to calculate optimum nutrient density to maximize profit margins, using nonlinear programming by predictive broiler performance. To demonstrate the influence of the broiler price could interact with nutrient density, the experiment aimed to define the quadratic equations for consumption and weight gain, based on modeling, to be applied to nonlinear programming, according to sex (male and female) in the starter (1 to 21 days), grower (22 to 42 days) and finisher phases (43 to 56 days). The experimental design was a randomized, totaling 6 treatments [energy levels of 2800, 2900, 3000, 3100, 3200 and 3300kcal AME/kg with constant nutrient : AME (Apparent Metabolizable Energy)] with 4 replicates and 10 birds per plot, using the program free download PPFR Excel workbook for feed formulation (http://www.foa.unesp.br/downloads/file_detalhes.asp?CatCod=4&SubCatCod=138&FileCod=1677). Data from this trial confirmed that there was a significant relationship between feed intake and total energy consumption of the diet, in which feed intake was increased or decreased simply to keep the amount of energy, with a constant rate of nutrient : AME. Therefore, the data support that if the essential dietary nutrients are kept in proportion to the energy density of the diet, according to the appropriate requirements (male / female) of broilers, the weight and feed conversion are significantly (P<0.05) favored by increasing the energy density of the diet. Thus, it enables the application of models for maximum profit (nonlinear formulation), to estimate the proportion of weight gain most appropriate according to the price paid by the market.

Bacterial oxycarotenoids added into broiler chickens ration modify breast meat color

Some markets around the world demand for well pigmented poultry products, what justifies the intentional use of synthetic additives in rations during breeding. The inconvenient for that practice most times is the elevated price of these products, beyond the argument of being chemicals, which may not be viable for some production systems. So, the aim of this experiment was to check on the ability of natural red oxycarotenoids produced by R. gelatinosus to modify broilers breast meat color. Bacterial biomass containing oxycarotenoids were added into Cobb broilers finisher diets at 0, 1.0, 2.0 and 3.0 g/kg and fed during 10 days (35-45 d). Five replicates (10 birds each) were performed. At days 35, 37, 39, 41, 43 and 45, 100 birds were slaughtered scalded, defeathered and eviscerated. Data on live and carcass weights were recorded. After cooling, color parameters of meat were determined on breast surface using a HunterLab XE Plus colorimeter calibrated with black and white standard tiles. Regression analysis was used for the investigation of relationships between days of consumption and biomass concentration on color values, live weight and carcass yield. Results showed that redness of breast meat was significantly influenced by the biomass concentration in a linear effect (P=0.0056) and also by the time of consumption, in a quadratic effect (P=0.0232). Days of consumption also affected yellowness and lightness with significant quadratic responses (P=0.0225 and P<0.0001, respectively). Birds live weight increased significantly during the application of the experimental diets (P<0.0001), showing no negative influence of the biomass administration. Also no deleterious effects derived from the application of the product were observed on carcass yield. So we concluded that R. gelatinosus oxycarotenoids present in the bacterial biomass can modify broilers breast meat color, performing as an alternative pigmenting additive in poultry production.

Accurate adjustment of feed formulation for broiler chickens on performance and lipid composition of meat

Most consumers consider the fat of chicken meat undesirable for a healthy diet, due to the high levels of saturated fatty acids and cholesterol. The purpose of this experiment was to investigate the influence of changes in dietary metabolizable energy level, associated with a proportional nutrient density variation, on broiler chickens performance and on the lipid composition of meat. Males and females Cobb 500 broilers were evaluated separately. Performance evaluation followed a completely randomized design with factorial 6x3 arrangement - six energy levels (2,800, 2,900, 3,000, 3,100, 3,200 and 3,300 kcal/kg) and three slaughter ages (42, 49 and 56 days). Response surface methodology was used to establish a mathematical model to explain live weight, feed intake and feed conversion behavior. Total lipids and cholesterol were determined in skinned breast meat and in thigh meat, with and without skin. For lipid composition analysis, a 3x3x2 factorial arrangement in a completely randomized design - three ration’s metabolizable energy levels (2,800, 3,000 and 3,300 kcal/kg), three slaughter ages (42, 49 and 56 days) and two sexes - was used. The reduction in the diet metabolizable energy up to close to 3,000 kcal/kg did not affect live weight but, below this value, the live weight decreased. Feed intake was lower when the dietary energy level was higher. Feed conversion was favored in a direct proportion to the increase of the energy level of the diet. The performance of all birds was within the range considered appropriate for the lineage. Breast meat had less total lipids and cholesterol than thigh meat. Thigh with skin had more than the double of total lipids of skinned thigh, but the cholesterol content did not differ with the removal of the skin, suggesting that cholesterol content is not associated with the subcutaneous fat. Intramuscular fat content was lower in the meat from birds fed diets with lower energy level. These results may help to define the most appropriate nutritional management. Despite the decrease in bird’s productive performance, the restriction of energy in broiler chickens feed may be a viable alternative, if the consumers are willing to pay more for meat with less fat.

Strategies to minimize heat stress in broilers chickens

The objective of this study was to evaluate the application of early age thermal conditioning (TC) and dietary electrolyte balances (DEB) to minimize the effect of heat stress. A total of 240 Cobb 500 1-d-old male broiler chicks was randomly allocated to 24 floor pens with six replicates per treatment in a 2x2 factorial arrangement (with and without TC and with and without DEB). Dietary treatments consisted: (T1) a traditional diet without TC; (T2) traditional diet with TC; (T3) with the application of dietary electrolyte and without TC and (T4) application of dietary electrolyte with TC. The thermal conditioning was conducted at 5 d of age (36°C for 24 h), only half of the batch (120 birds). After this period, all birds were transferred to boxes of 1.5 x 3m (10 birds / box), with wood shavings reused as litter. Chicks were exposed to chronic heat stress (32°C) for 6 h from 35 to 39 d of age, in all treatments, being electronically monitored the temperature and humidity of the microclimate of the birds. Feed and water were provided ad libitum, even during periods of stress. Were measured performance data (weight gain, feed intake and feed conversion) and mortality rate. Data were subjected to ANOVA using the GLM procedures of SAS. The results of this study demonstrated no interaction effects of all evaluated parameters (performance and mortality). Therefore, no synergism occurred when both strategies (TC and DEB) were applied. Only the first week there was prejudice on broiler performance with the application of the TC and apparently the technique of early age thermal conditioning no improve the resistance of broilers to chronic heat stress. On the other hand, DEB had significantly favorable effects (P<0.05) on performance and minimized mortality immediately after application of the TC.

Serologic and molecular diagnostic and bioassay in mice for detection of Toxoplasma gondii in free ranges chickens from Pantanal of Mato Grosso do Sul

Holsback L., Pena H.F.J., Ragozo A., Lopes E. G., Gennari S. M. & Soares R. M. 2012. Serologic and molecular diagnostic and bioassay in mice for detection of Toxoplasma gondii in free range chickens from Pantanal of Mato Grosso do Sul. Pesquisa Veterinaria Brasileira 32(8): 721-726. Setor de Veterinaria e Producao Animal, Universidade Estadual do Norte do Parana, Campus Luiz Meneghel, Rodovia BR 369 Km 54, Bandeirantes, PR 86360-000, Brazil. E-mail: lhsfertonani@uenp.edu.br The aim of this study was to investigate the occurrence of Toxoplasma gondii and compare the results obtained in the Modified Agglutination Test (MAT), Polimerase Chain Reaction (PCR) and bioassay in mice. In order to accomplish this, 40 free-range chickens from eight farms in neighboring areas to the Pantanal in Nhecolandia, Mato Grosso do Sul, were euthanized and blood samples, brain and heart were collected. The occurrence of anti-T. gondii antibodies found in chickens was 67.5% (27 samples), considering as a cutoff point the dilution 1:5. Among the samples analyzed, 7 (25.9%) were positive in the dilution 1: 5, 3 (11.1%) in 1: 10, 2 (7.4%) in 1: 20, 3 (11.1%) in 1: 320, 1 (3.7%) in 1: 640, 3 (11.1%) in 1: 1280, 2 (7.4%) in 1: 2560, 4 (14.8%) in 1: 5120 and 2 (7.4%) in 1: 10.240. From the mixture of tissue samples (brain and heart) from the chickens analyzed, 16 (40%) presented electrophoretic bands compatible with T. gondii by PCR (gene B1). In the comparison of techniques, 59.26% positivity in PCR was revealed among animals that were seropositive in MAT (cutoff 1: 5). From 141 inoculated mice, six (4.44%) died of acute toxoplasmosis between 15 and 23 days after inoculation. Surviving mice were sacrificed at 74 days after inoculation, and a total of 28 cysts were found in the brains of 10 distinct groups. From the seropositive hens, 27 bioassays were performed and 11 (40.7%) isolates were obtained. A greater number of isolations happened in mice that were inoculated with tissues from chickens that had high titers for anti-T. gondii antibodies. Chronic infection in mice was observed in nine groups (33.3%) from five different properties. Among the surviving mice, 25.6% were positive for T. gondii in MAT (1: 25). From mice positive in PCR, 87.5% were also positive in MAT. Among the PCR-negative mice, 5.2% were positive for T. gondii in MAT. It can be concluded through this study that the occurrence of infecton by T. gondii in the rural properties studied was high, that PCR directed to gene B1 does not confirm the viability of the parasite, but it can be used as a screening method for the selection of chickens infected by T. gondii, that the animals with titer greater than 10 must be prioritized for the selection of animals for bioassay, since for them, the chances of isolating the parasite are greater and that seroconversion in experimentally infected mice is not a good indicator for isolating the agent.

Acute heat stress impairs performance parameters and induces mild intestinal enteritis in broiler chickens: Role of acute hypothalamic-pituitary-adrenal axis activation

Studies on the environmental consequences of stress are relevant for economic and animal welfare reasons. We recently reported that long-term heat stressors (31 +/- 1 degrees C and 36 +/- 1 degrees C for 10 h/d) applied to broiler chickens (Gallus gallus domesticus) from d 35 to 42 of life increased serum corticosterone concentrations, decreased performance variables and the macrophage oxidative burst, and produced mild, multifocal acute enteritis. Being cognizant of the relevance of acute heat stress on tropical and subtropical poultry production, we designed the current experiment to analyze, from a neuroimmune perspective, the effects of an acute heat stress (31 +/- 1 degrees C for 10 h on d 35 of life) on serum corticosterone, performance variables, intestinal histology, and peritoneal macrophage activity in chickens. We demonstrated that the acute heat stress increased serum corticosterone concentrations and mortality and decreased food intake, BW gain, and feed conversion (P < 0.05). We did not find changes in the relative weights of the spleen, thymus, and bursa of Fabricius (P > 0.05). Increases in the basal and the Staphylococcus aureus-induced macrophage oxidative bursts and a decrease in the percentage of macrophages performing phagocytosis were also observed. Finally, mild, multifocal acute enteritis, characterized by the increased presence of lymphocytes and plasmocytes within the lamina propria of the jejunum, was also observed. We found that the stress-induced hypothalamic-pituitary-adrenal axis activation was responsible for the negative effects observed on chicken performance and immune function as well as for the changes in the intestinal mucosa. The data presented here corroborate with those presented in other studies in the field of neuroimmunomodulation and open new avenues for the improvement of broiler chicken welfare and production performance.

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espirito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espirito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo (R), Wama Diagnostica, Sao Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of >= 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer >= 1:16) and 38.8% (198/510) by MAT (titer >= 1:25); concordance between IHAT and MAT was 81.6% (kappa index = 0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64(75%) seropositive (IHAT titers >= 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies. (C) 2012 Elsevier B.V. All rights reserved.