Nicarbazin Residue in Tissues from Broilers Reared on Reused Litter Conditions.

Nicarbazin (NCZ) is a worldwide used anticoccidial in poultry farming to avoid coccidiosis disease when chickens are reared on conventional poultry litter. If proper dosage and withdrawal time are not followed, the component dinitrocarbanilide (DNC) of NCZ may be present in chicken tissues, therefore posing a risk to consumers if the residues are above 200 µg/kg. Litter reuse is a common and important practice in commercial chicken production. Literature is lacking about the influence of litter reuse on DNC deposition in chicken tissues and its impact on food safety. We aimed to evaluate DNC residues in breast and liver by LC-MS/MS from broilers from an experiment with 10 consecutive flocks during 2 years. The experiment included three treatments containing NCZ in the diet (T1 = 125 mg/kg, 1−21 d; T2 = 125 mg/kg, 1−32 d; T3 = 40 mg/kg, 1−32 d). DNC residues in chicken breast at 21 d in T1 ranged from 648.8−926 µg/kg, at 32 d in T2 and T3 varied, respectively, from 232−667 µg/kg and 52−189 µg/kg. Regarding liver, DNC residues at 21 days in T1 ranged from 11,754−15,281 µg/kg, at 32 days in T2 and T3 varied, respectively, from 10,168−15,021 µg/kg and 2899−4573 µg/kg. When NCZ was withdrawn from feed, DNC residues dropped to

Effect of gaseous ozone application during chilling on microbial and quality attributes of pig carcasses.

Ozone application has been suggested as an additional measure to the slaughter animals under hygiene programs. In this study, we determined the efficacy of gaseous ozone applied to pig carcasses during chilling (16 h at 2-5°C). Forty carcasses were allocated to each treatment: control, without ozone application (T1) and 5 ppm gaseous ozone application (T2), divided in two 4-h periods. The carcasses were sampled before and after chilling. The average counts of total aerobic mesophilic (TAM) bacteria before chilling were not different (p = 0.55) between T1 and T2. In turn, after chilling, the ozone-treated carcasses had significantly reduced about 0.4 colony-forming units (CFU)/cm2 of TAM counts (p < 0.001) than the control carcasses. No significant reduction was observed in the number of carcasses positive for Listeria sp. and Escherichia coli after gaseous ozone treatment; while a tendency (p = 0.08) of lower number of Salmonella positive carcasses in T2 was observed. Common macrorestriction (pulsed-field gel electrophoresis) patterns of S. enterica were observed in the carcasses before and after chilling. Pork samples from treated and untreated carcasses with ozone showed no lipid oxidation or altered color and pH. The results indicate that the gaseous ozone in the tested protocol is effective in reducing TAM populations, but not effective in decreasing the number of carcasses positive for E. coli and Listeria sp. Regarding Salmonella, the tendency of positive carcasses reduction may encourage further studies by testing other protocols of gaseous ozone application inside the chilling chamber.

Performance, carcass and meat quality of pigs submitted to immunocastration and different feeding programs.

The feed restriction of pigs aims to improve the nutrient utilization of the diet, reduce carcass fat and increase lean meat. Immunocastration is a painless, reversible technique that improves animal welfare, decreases boar taint in meat, leading to lower fat deposition and increases lean meat. This study aimed at evaluating either feeding (restricted) programs (FP) and castration categories (surgically castrated × immunocastration) in terms of pig performance, carcass traits and meat quality. Castration category and FP did not influence marbling, initial pH and drip loss in loin and ham. However, immunocastrated pigs presented lighter loins and hams with lower ultimate pH, compared to the surgically castrated pigs. The FP with lower feed volume resulted in higher ultimate pH. Immunocastrated males had a higher live weight than those surgically castrated. Also, the more the feed supply decreased, the better the feed conversion. Immunocastrates had lower carcass yields, however these pigs had their backfat thickness decreased and lean meat increased in the carcass, without affecting the loin depth.

Detection of p-Nitroaniline Released from Degradation of 4,4'-Dinitrocarbanilide in Chicken Breast during Thermal Processing.

The diphenylurea 4,4'-dinitrocarbanilide (DNC) is the residue of concern left in edible tissues of broilers fed diets containing the anticoccidial nicarbazin. When chicken meat is submitted to thermal processing, p-nitroaniline (p-NA) is expected from DNC degradation. This work aimed at evaluating whether thermal processing of DNC-containing chicken meat induces p-NA appearance. First, a hydrolysis assay was performed in aqueous solutions at 100 °C in different pH, confirming that DNC cleavage yields p-NA. Then a novel LC-MS/MS method was used to detect traces of this aromatic amine in DNC-containing chicken breast fillets subjected to cooking methods. Our evidence showed p-NA occurrence in such chicken meat samples, which corroborated results from hydrolysis assay. The p-NA appearance in fillets was rather discrete during boiling treatment, but its concentration became pronounced over time for grilling, frying, and roasting, achieving respectively 326.3, 640.0, and 456.9 µg/kg. As far as we are concerned, no other research identified degradation products from DNC residue in heat-processed chicken fillets. Therefore, this study leads to additional approaches to assess impacts on food safety.

Risk factors associated with pig pre-slaughtering losses.

The incidence of non-ambulatory non-injured (NANI), non-ambulatory injured (NAI) and dead pigs on-arrival at three Brazilian slaughterhouses were evaluated in 37,962 pigs to identify risk factors linked to them, besides carcass bruises and limb fractures. Total pre-slaughtering losses were 1.18%, in which NAI (0.39%) and NANI (0.37%) incidences contributed the most. A positive relation between on farm steeper ramp slope >20° and the incidence of NAI, NANI and dead pigs at unloading was found. Farm size, pigs/pen, enthalpy at loading, transportation time, truck loading order, muscle thickness and carcass weight, were identified as risk factors for pre-slaughtering losses. Loading procedures influenced the occurrence of limb fractures and bruises (which are a welfare issue and should be reduced). Therefore, personal training on pre-slaughter handling is essential to reduce the risk factors to improve animal welfare and avoid losses during the pre-slaughter process.

Determination of ractopamine residue in tissues and urine from pig fed meat and bone meal.

In many countries, ractopamine hydrochloride (RAC) is allowed to be used in animal production as a ß-agonist, which is an energy repartitioning agent able to offer economic benefits such as increased muscle and decreased fat deposition, feed conversion improvement and an increase in average daily weight gain. However, some countries have banned its use and established strict traceability programmes because of pharmacological implications of ß-agonist residues in meat products. In Brazil, commercial RAC is controlled (5-20 mg kg-1) and only added to pig diet during the last 28 days before slaughter. However, the control is more difficult when co-products, like meat and bone meal (MBM), which can be produced from RAC treated animals, are part of the feed composition. Therefore, a study was undertaken to evaluate the presence of RAC residue concentrations in urine and tissues of gilts (n = 40) in four dietary groups: 0%, 7%, 14% and 21% (w/w) of MBM-containing RAC (53.5 µg kg-1). The concentration of RAC residues in MBM, pig tissues and urine was determined by LC-MS. Low RAC concentrations were detected in muscle, kidney, liver and lungs (limit of detection = 0.15, 0.5, 0.5 and 1.0 µg kg-1, respectively); however, no RAC residues were quantified above the limit of quantification (0.5, 2.5, 2.5 and 2.5 µg kg-1, respectively). In urine, the RAC concentration remained below 1.35 µg L-1. These data suggest that MBM (containing 53.5 µg kg-1 RAC) added to diet up to 21% (w/w) could hamper the trade where RAC is restricted or has zero-tolerance policy.

Application of Enzyme Digestion and Deconjugation Followed by Quick, Easy, Cheap, Effective, Rugged, Safe Extraction and Liquid Chromatography-Tandem Mass Spectrometry Methodology To Determine Ractopamine Residue in Pork.

A new methodology is proposed for ractopamine residue analysis in pork. It consists of enzyme-mediated digestion and deconjugation steps; modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction; and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In brief, the samples were digested with protease and then deconjugated with ß-glucuronidase enzyme; they were then subjected to extraction and cleanup by QuEChERS and underwent sequential analysis by LC-MS/MS. The method performance was evaluated in accordance to the validation guidelines regulated by the Brazilian Ministry of Agriculture and Food Supply. The limit of detection was 0.15 µg/kg and limit of quantification was 0.5 µg/kg. When the method was applied to real samples, ractopamine residue was found in concentrations (up to 7.86 µg/kg) below international recommendation limits up to 10 µg/kg. The method is sensitive, accurate, quick, simple, and suitable for routine analysis; therefore, it is a monitoring tool that may be adopted by laboratories to achieve compliance levels.

Degradation of 4,4'-Dinitrocarbanilide in Chicken Breast by Thermal Processing.

Nicarbazin is one of the major anticoccidials used in broiler feeds. The compound 4,4'-dinitrocarbanilide (DNC) is the marker residue of concern left from nicarbazin in chicken meat. The effect of thermal processing on DNC content accumulated in chicken breast was assessed, and samples were analyzed by liquid chromatography-tandem mass spectrometry. Five conventional cooking methods were evaluated: boiling, grilling, microwaving, frying, and roasting. To ensure DNC in meat, broilers were fed nicarbazin without withdrawal period. All heating methods surpassed the 70 °C end point core temperature in chicken breast. Maximum DNC degradation was reached at 10 min for boiling, at 30 min for grilling, and at 2 min for microwaving, and no further reduction was observed for longer thermal processing time. Boiling was more efficient in reducing DNC (69%). Grilling, microwaving, and frying achieved on average 55% of degradation. The outcomes reported herein may be considered in decision-making regarding further review of maximum residue limits.

Ractopamine analysis in pig kidney, liver and lungs: A validation of the method scope extension using QuEChERS as a sample preparation step.

Ractopamine has been allowed by some countries as a repartitioning additive in pig diet, since it promotes protein synthesis and fat lipolysis. Most regulatory agencies only propose the ractopamine assessment in meat, kidney, liver and fat. Aiming at contributing to the scarcity data regarding this analyte in pig lungs, we extended the scope of a LC-MS method to evaluate pig offals. Homogenized tissue samples were extracted by a QuEChERS procedure; following by clean up steps and further tandem mass spectrometry determination. Method performance was evaluated through specificity, recovery, linearity, reproducibility, repeatability, decision limit (CCα), and detection capability (CCß), in accordance to the Commission Decision 2002/657/EC. Regression coefficients (R2) between 0.994 and 0.999 were achieved for kidney, liver and lungs. Recoveries ranged from 92.0 to 127%. CCα and CCß values ranged from 3.65 to 4.86 µg kg-1, and from 6.27 to 7.21 µg kg-1, respectively. These values were under the maximum residue limits suggested by Codex Alimentarius, which are 90 and 40 µg kg-1 for kidney and liver, respectively. When applied to real samples up to 22.5, 92 and 1003 µg kg-1 of ractopamine residues were detected in pig liver, kidney and lungs, respectively. The results allowed concluding that the proposed analytical method is capable to detect ractopamine residues in all evaluated matrices. Therefore, it can be successfully applied and used as a routine method in laboratories of residue analysis.

Current research, regulation, risk, analytical methods and monitoring results for nicarbazin in chicken meat: A perspective review.

This review presents up-to-date information about current research on nicarbazin, one of the most used anticoccidials in poultry production. The focus is to elucidate regulation concerning nicarbazin, limits for its residues in food, how maximum residue limits in different countries are calculated regarding edible chicken tissues and the possible implications in human health. Analytical methods to extract and quantify this residue, expressed as dinitrocarbanilide (DNC) are presented and discussed, including qualitative screening and quantitative/confirmatory analytical methods. Monitoring results and occurrence of DNC residues in chicken meat are discussed. Additionally, the causes of eventual chicken meat contamination and possible solutions to reduce or eliminate DNC residue in tissues are also presented. The paper concludes with perspectives, the current state of DNC residue analysis and suggestions for future research, especially considering the gap in the study of residue recycling effect due to continuous chicken litter use.

Egg quality assessment at different storage conditions, seasons and laying hen strains / Avaliação da qualidade do ovo em diferentes condições de armazenamento, estações do ano e linhagens de poedeiras

ABSTRACT Eggs are one of nearly perfect protein foods, offering nutrients of great biological value. However, during storage, egg albumen and yolk components may alter and deteriorate egg quality. Therefore, the aim of this work was to assess egg quality during 9-week storage. Parameters such as Haugh unit, weight loss, egg width and length, specific gravity, yolk and albumen dimensions and their pH were evaluated weekly. A total of 270 eggs (n=5) collected from two different hen strains were evaluated under room (20 to 35 °C in summer; 11.2 to 29.7 °C in autumn) and refrigerated (0 to 5 °C in summer; -3.1 to 6.5 °C in autumn) temperatures. For storage time, an unfolding analysis was accomplished by regression analysis using orthogonal polynomials. As a second approach, principal component analysis (PCA) was performed in order to assess correlations among quality parameters on storage conditions and laying hen strains. By the end of the 9-week storage period, eggs kept under refrigeration presented similar quality parameters to eggs stored at room temperature for only 3 weeks. In contrast, eggs kept at room temperature presented faster degradation from week 1 to 5. No differences on egg quality parameters were noticed between white and brown shells eggs. PCA suggests that better egg quality (first week) was associated mainly with higher egg weight and its specific gravity, Haugh unit and albumen height. Eggs stored at room temperature should be consumed in 2 weeks or refrigerated until 8 weeks, preserving internal quality from farm to retail.

RESUMO Os ovos são um dos alimentos que oferecem nutrientes de grande valor biológico. Entretanto, no armazenamento, alguns componentes da clara e gema podem se alterar, deteriorando a qualidade dos ovos. Assim, este trabalho objetivou avaliar semanalmente parâmetros de qualidade como unidade Haugh, perda de peso, largura/comprimento do ovo, densidade específica, dimensões da clara/gema e pH durante 9 semanas de armazenamento. Foram coletados 270 ovos de duas linhagens de poedeiras e avaliados a temperatura ambiente (20 a 35 °C no verão; 11,2 a 29,7 °C no outono) e refrigerada (0 a 5 °C no verão; -3,1 a 6,5 ºC no outono). Para as condições de armazenamento, foi realizada uma análise de desdobramento por regressão por polinômios ortogonais. Ainda, análise de componentes principais (PCA), objetivou avaliar correlações entre parâmetros de qualidade em diferentes condições de armazenamento e linhagens de poedeira. Após 9 semanas de armazenamento, os ovos refrigerados apresentaram qualidade similar àqueles armazenados a temperatura ambiente por 3 semanas. Porém, sem refrigeração, os ovos apresentaram uma degradação mais rápida da semana 1 a 5. Não foram observadas diferenças nos parâmetros de qualidade dos ovos entre as diferentes colorações de casca (linhagens de poedeiras). PCA sugere que a melhor qualidade dos ovos (primeira semana) foi associada principalmente com valores elevados de peso e densidade específica do ovo, unidade Haugh e altura do albúmen. Ovos armazenados a temperatura ambiente devem ser consumidos até duas semanas ou mantidos sob refrigeração até 8 semanas, preservando a qualidade interna desde a granja até o armazenamento no varejo.

Effects of biodiesel made from swine and chicken fat residues on carbon monoxide, carbon dioxide, and nitrogen oxide emissions.

The effects of two alternative sources of animal fat-derived biodiesel feedstock on CO2, CO, NOx tailpipe emissions as well as fuel consumption were investigated. Biodiesel blends were produced from chicken and swine fat waste (FW-1) or floating fat (FW-2) collected from slaughterhouse wastewater treatment processes. Tests were conducted in an unmodified stationary diesel engine operating under idling conditions in attempt to simulate slow traffic in urban areas. Significant reductions in CO (up to 47% for B100; FW-2) and NOx (up to 20% for B5; FW-2 or B100; FW-1) were attained when using biodiesel fuels at the expense of 5% increase in fuel consumption. Principal component analysis (PCA) was performed to elucidate possible associations among gas (CO2, CO, and NOx) emissions, cetane number and iodine index with different sources of feedstock typically employed in the biodiesel industry. NOx, cetane number and iodine index were inversely proportional to CO2 and biodiesel concentration. High NOx emissions were reported from high iodine index biodiesel derived especially from forestry, fishery and some agriculture feedstocks, while the biodiesel derived from animal sources consistently presented lower iodine index mitigating NOx emissions. The obtained results point out the applicability of biodiesel fuels derived from fat-rich residues originated from animal production on mitigation of greenhouse gas emissions. The information may encourage practitioners from biodiesel industry whilst contributing towards development of sustainable animal production. IMPLICATIONS: Emissions from motor vehicles can contribute considerably to the levels of greenhouse gases in the atmosphere. The use of biodiesel to replace or augment diesel can not only decrease our dependency on fossil fuels but also help decrease air pollution. Thus, different sources of feedstocks are constantly being explored for affordable biodiesel production. However, the amount of carbon monoxide (CO), carbon dioxide (CO2), and/or nitrogen oxide (NOx) emissions can vary largely depending on type of feedstock used to produce biodiesel. In this work, the authors demonstrated animal fat feasibility in replacing petrodiesel with less impact regarding greenhouse gas emissions than other sources.

Development of a readily applied method to quantify ractopamine residue in meat and bone meal by QuEChERS-LC-MS/MS.

A QuEChERS method of ractopamine (RCT) residue detection in swine meat and bone meal (MBM) samples was demonstrated. Samples were hydrolyzed with protease and ß-glucuronidase prior to QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) extraction and clean-up. Samples were analyzed in a Liquid Chromatography (equipped with ACE 5 C18 column under gradient elution) coupled with a triple quadrupole mass spectrometer operating in positive electrospray ionization mode (using multiple reaction monitoring, MRM). The method was validated for its specificity, decision limit (CCα), detection capability (CCß), recovery, repeatability, reproducibility, linearity, limits of detection (LODs), quantification (LOQs), and stability according to international guidelines (European Commission Decision 2002/657/EC). Recoveries ranged from 96.3 to 107.0%. Repeatability and reproducibility showed both RSD<5.7% and 3.1%, respectively. LODs and LOQs were 1.91 and 6.36 ppb, respectively. CCα and CCß values were 1.91 and 2.37 ppb, respectively. RCT showed good stability for spiked samples and real samples when the concentration was higher, otherwise at lower concentration stability was lower. The proposed method can be successfully applied on a regular basis for the determination of RCT in MBM, demonstrating the usefulness of the method as a tool for compliance monitoring in regulatory laboratories.

Changes in lipid, fatty acids and phospholipids composition of whole rice bran after solid-state fungal fermentation.

The aim of this study was to evaluate fermented rice bran phospholipids, lipids and fatty acid content in a fermentation solid system with Rhizopus oryzae fungus. For this, aliquots were withdrawn every 24h over 120 h. The content of phospholipids was determined by colorimetric method. Esterified fatty acids were separated by gas chromatography, then identified and quantified. The total lipids from fermented rice bran (FB) decreased from 20.4% to 11.2% in the range between 0 h and 120 h of fermentation while phospholipid contents were increased up to 2.4 mg P g(lipid)(-1). In fermented bran, oleic, palmitic and linoleic acids prevailed, with a decrease in saturated fatty acids (20%) and increase in the unsaturated ones (5%). This study showed that rice bran fermentation with R. oryzae can be applied to the production of phospholipids altering the saturated to unsaturated fatty acid ratio.

Improved enzymatic production of phenolated acylglycerols through alkyl phenolate intermediates.

A novel approach is reported for the synthesis of dihydrocaffoylated glycerols that consists of two steps: enzymatic synthesis of octyl dihydrocaffeate (as a synthetic intermediate) from octanol and dihydrocaffeic acid, and enzymatic interesterification of triacylglycerols with octyl dihydrocaffeate. Due to the good compatibility of the intermediate with triacylglycerols, an improved volumetric productivity [147 mol h(-1)(kg Novozym 435)(-1)] and high enzyme specific activity [up to 9.6 µmol(-1) min(-1)(g Novozym 435)(-1)] have been obtained.