Occurrence and bioaccumulation study of PCDD and PCDF from mineral feed additives

Occurrence of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) was evaluated in sepiolite as a widely employed binder and anti-caking agent for animal feed. Also, naturally contaminated kaolinitic clay was used for comparative purposes. Since sepiolite shows remarkable adsorption properties, particular interest was paid to the extraction steps as they become critical for the final determination of these pollutants in such matrixes. Furthermore, classical Soxhlet extraction using different extracting strategies as well as acid treatment were carried out with simultaneous liquid-liquid extraction. Results obtained depended on the extraction procedure applied. Acid treatment or Soxhlet extraction using a mixture of toluene:ethanol as solvent allowed to reach the minimum requirements of recovery rates. However, Soxhlet extraction using a mixture cyclohexane:toluene as extracting solvent did not allow to comply with minimum specifications for recovery. Significant differences were obtained in TEQ units when acid treatment was applied in comparison to Soxhlet extraction. This fact can be explained because the use of drastic acid conditions allows removing strongly adsorbed analytes which can be uniquely extracted after a total destruction of the crystalline structure of sepiolite. On the contrary, Soxhlet extraction was not able to destroy the structure of sepiolite and as a consequence the PCDDs/Fs were strongly adsorbed in the internal structure of the mineral. From biological point of view the availability of these toxicants constitutes a critical aspect playing an important role in the final decision choosing particular analytical procedures. Then, acid conditions in the digestive tract should be taken into account. In this scenario, a bioaccumulation study was conducted to evaluate the transference of PCDDs/PCDFs from the sepiolite into the animal tissues when fed with feed containing sepiolite. To this end, chickens were used as a model to examine the bioavailability of PCDDs/PCDFs. Four groups of chickens were exposed through their diet to a control feed, feed with 3% w/w sepiolite as additive, feed contaminated with PCDDs/PCDFs at concentration around 2.8 pg WHO-TEQ/g and feed with 2% of a contaminated kaolinitic clay (460 pg TEQ/g mineral). Livers of the four studied groups were analyzed throughout the exposure period. Results of this trial showed that the performance of broilers was not affected by the presence of dioxins at levels tested, and chickens did not show any abnormal behaviour. Dioxins intentionally added to the diet were absorbed and accumulated in the liver in a significant manner, whereas the PCDDs/Fs from sepiolite were not available for chickens since livers from broilers fed 3% sepiolite presented similar WHO-TEQ values than those from broilers fed control diet.

Overfishing of small pelagic fishes increases trophic overlap between immature and mature striped dolphins in the Mediterranean Sea

The interactions among diet, ecology, physiology, and biochemistry affect N and C stable isotope signatures in animal tissues. Here, we examined if ecological segregation among animals in relation to sex and age existed by analyzing the signatures of delta15N and delta13C in the muscle of Western Mediterranean striped dolphins. Moreover, we used a Bayesian mixing model to study diet composition and investigated potential dietary changes over the last two decades in this population. For this, we compared isotope signatures in samples of stranded dolphins obtained during two epizootic events occurring in 1990 and 2007-2008. Mean delta13C values for females and males were not significantly different, but age-related variation indicated delta13C enrichment in both sexes, suggesting that females and males most likely fed in the same general areas, increasing their consumption of benthic prey with age. Enrichment of delta15N was only observed in females, suggesting a preference for larger or higher trophic level prey than males, which could reflect different nutritional requirements. delta13C values showed no temporal variation, although the mean delta15N signature decreased from 1990 to 2007-2008, which could indicate a dietary shift in the striped dolphin over the last two decades. The results of SIAR indicated that in 1990, hake and sardine together contributed to 60% on the diet of immature striped dolphins, and close to 90% for mature striped dolphins. Conversely, the diet of both groups in 2007-2008 was more diverse, as hake and sardine contributed to less than 40% of the entire diet. These results suggest a dietary change that was possibly related to changes in food availability, which is consistent with the depletion of sardine stocks by fishing.

A radiochemical method for carbamoyl-phosphate synthetase I: application to rats fed a hyperproteic diet

A method for the measurement of carbamoyl-phosphate synthetase I activity in animal tissues has been developed using the livers of rats under normal and hyperproteic diets. The method is based on the incorporation of 14C-ammonium bicarbonate to carbamoyl-phosphate in the presence of ATP-Mg and N-acetyl-glutamate. The reaction is stopped by chilling, lowering the pH and adding ethanol. Excess bicarbonate is flushed out under a gentle stream of cold CO2. The only label remaining in the medium was that incorporated into carbamoyl-phosphate, since all 14C-CO2 from bicarbonate was eliminated. The method is rapid and requires only a low pressure supply of CO2 to remove the excess substrate. The reaction is linear up to 10 min using homogenate dilutions of 1:20 to 1:200 (w/v). Rat liver activity was in the range of 89±8 nkat/g. Hyperproteic diet resulted in a significant 1.4-fold increase. The design of the method allows for the processing of multiple samples at the same time, and incubation medium manipulation is unnecessary, since the plastic incubation vial and its contents are finally counted together.

A radiochemical method for carbamoyl-phosphate synthetase I: application to rats fed a hyperproteic diet

A method for the measurement of carbamoyl-phosphate synthetase I activity in animal tissues has been developed using the livers of rats under normal and hyperproteic diets. The method is based on the incorporation of 14C-ammonium bicarbonate to carbamoyl-phosphate in the presence of ATP-Mg and N-acetyl-glutamate. The reaction is stopped by chilling, lowering the pH and adding ethanol. Excess bicarbonate is flushed out under a gentle stream of cold CO2. The only label remaining in the medium was that incorporated into carbamoyl-phosphate, since all 14C-CO2 from bicarbonate was eliminated. The method is rapid and requires only a low pressure supply of CO2 to remove the excess substrate. The reaction is linear up to 10 min using homogenate dilutions of 1:20 to 1:200 (w/v). Rat liver activity was in the range of 89±8 nkat/g. Hyperproteic diet resulted in a significant 1.4-fold increase. The design of the method allows for the processing of multiple samples at the same time, and incubation medium manipulation is unnecessary, since the plastic incubation vial and its contents are finally counted together.

Stable C and N isotope concentration in several tissues of the loggerhead sea turtle Caretta caretta from the western Mediterranean and dietary implications

The isotopic concentrations of carapace scutes, skin, muscle and blood of loggerhead sea turtles (Caretta caretta) from the Balearic Archipelago were analysed to investigate the pattern of variation between tissues and to assess the position of this species in the trophic webs of the Algerian Basin. Skin showed higher δ13C values than muscle or carapace scutes and these showed higher values than blood. Conversely, muscle showed higher δ15N values than skin, skin showed higher values than blood and blood showed higher values than carapace scutes. Dead and live sea turtles from the same habitat did not differ in the concentration of stable isotopes. However, some of the tissues of the turtles caught in drifting longlines in the oceanic realm showed higher δ13C values than those from the turtles caught by hand or in trammel nets over the continental shelf, although they did not differ in the δ15N. Comparison of the concentration of stable isotopes in the turtles with that of other species from several areas of the Algerian Basin revealed that they consumed planktonic prey and that the trophic level of the sea turtles was higher than that of carnivorous cnidarians but lower than that of zooplanktophagous fish and crustaceans.

Comparison of fatty acid profiles of edible meat, adipose tissues and muscles between cocks and capons

The effect of caponisation on fat composition by parts (wing, breast, thigh, and drumstick) and tissues (skin, subcutaneous adipose tissue, intermuscular adipose tissue and muscle) was examined in the present study and fatty acid profiles of abdominal fat and edible meat by parts and tissue components were determined. The sample was made up of twenty-eight castrated and twenty male Penedesenca Negra chicks reared under free-range conditions and slaughtered at 28 wk of age; the birds were castrated at four or eight weeks. Caponisation significantly increased (P < 0.01) the chemical fat content in all parts (16.31% to 37.98% in breast; 21.98% to 34.13% in wing; 21.09% to 49.57% in thigh; 14.33% to 24.82% in drumstick) and led to minor modifications in fat haracteristics, particularly in the thigh and the drumstick, where the unsaturated vs. saturated fatty acid ratio increased from 1.31 to 1.76 ( P < 0.01) and from 1.48 to 2.07 (P < 0.01), respectively. Delaying the age of castration from 4 to 8 weeks increased this ratio by 0.35 in the edible meat. Even though the profile of the abdominal fat is less saturated in capons, all changes occurring on fat quality after caponisation indicate that increased fatness after castration does not imply worse fat nutritional properties.

Comparison of carcass composition by parts and tissues between cocks and capons

The effect of caponisation on carcass composition by parts and tissues was examined. Twenty-eight castrated and twenty male Penedesenca Negra chicks reared under free-range conditions were slaughtered at 28 weeks of age. The birds were castrated at 4 or 8 weeks. The left sides of the carcasses were quartered (wing, breast, thigh and drumstick), and the parts dissected into the tissue components (skin, subcutaneous fat, intermuscular fat, muscle, bone and tendons). Capons showed more abdominal, intermuscular and subcutaneous fat than the cocks, both at the same slaughter age and at the same weight. The breast and thigh were heavier in the capons than in the cocks. However, the whole muscle mass in the breast was increased by caponisation. This favourable effect was achieved at the expense of decreasing the carcass yield. The age of castration up to 8 weeks did not affect the carcass composition of the parts and tissues.

Mitochondrial DNA damage and animal longevity: insights from comparative studies

Chemical reactions in living cells are under strict enzyme control and conform to a tightly regulated metabolic program. However, uncontrolled and potentially deleterious endogenous reactions occur, even under physiological conditions. Aging, in this chemical context, could be viewed as an entropic process, the result of chemical side reactions that chronically and cumulatively degrade the function of biological systems. Mitochondria are a main source of reactive oxygen species (ROS) and chemical sidereactions in healthy aerobic tissues and are the only known extranuclear cellular organelles in animal cells that contain their own DNA (mtDNA). ROS can modify mtDNA directly at the sugar-phosphate backbone or at the bases, producing many different oxidatively modified purines and pyrimidines, as well as single and double strand breaks and DNA mutations. In this scenario, natural selection tends to decrease the mitochondrial ROS generation, the oxidative damage to mtDNA, and the mitochondrial mutation rate in long-lived species, in agreement with the mitochondrial oxidative stress theory of aging.

Residues of b-lactams and quinolones in tissues and milk samples. Confirmatory analysis by liquid chromatography-mass spectrometry

The aim of this work is to optimize and validate methods for the multiresidue determination of series of families of antibiotics as quinolones, penicillins and cephalosporins included in European regulation in food samples using LC-MS/MS. Different extraction techniques and clean-up applied to antibiotics in meat were compared. The quality parameters were established according with EU guideline. The developed method was applied to 49 positive raw milk samples from animal medicated with different antibiotics; the 63% of the analyzed samples were found to be compliant. ___________________________________________________________________________________________

Determination and characterization of quinolones in foodstuffs of animal origin by CE-UV, LC-UV, LC-Fl, LC-MS and LC-MS/MS

In this work methods for the multiresidue determination of the series of quinolones include in the European regulation in food of animal origin are de veloped and validated in line with Commission Decision 2002/657/EC in terms of linearity, decision limit, capability detection, precision and stability. Mult iresidue methods were established to allow the determination of quinolones covered by EU legislation in 2377/90/EC in muscle of chicken, turkey, pig and cow, plasma of cow and pig, liver of pig and milk of cow. First an extraction step was optimized and a SPE step was applied to clean!up and preconcentrate quinolones prior to their separation by CE or LC and determination by CE!UV, LC!UV, LC!Fl, LC!MS with different ion sources (ESI ,ApCI) and different mass analyser (Q, ToF) and LC!E SI!QqQ tandem mass spectrometry. The limits of quantification obtained are always lower than Maxim um Residue Limit (MRL) established by EU for quinolones in animal products and they can be applied to the control of quinolones in foodstuffs of animal origin . Finally the proposed methods were applied to determine quinolones in samples of turkey and pig muscle, pig plasma and milk of cow. Excellent quality parameters and reduced time of analysis were obtained when LC!ESI!MS/MS is used, although the others techniques presented too satisfactory results.

Efectos del gen halotano sobre el bienestar animal, la calidad de carne y la productividad en porcinos

El Centro de Tecnología de la Carne fue responsable de tres de los objetivos fundamentales de este proyecto. Los experimentos consistieron en el sacrificio de dos lotes de cerdos que se diferenciaban en su genotipo halotano y en la línea paterna, que fueron sometidos a distintos tratamientos ante-mortem (transporte y espera largos versus transporte y espera cortos). En el primer lote, se compararon animales libres del gen con heterocigotos que procedían de una línea paterna Pietrain o una línea híbrida Large White x Pietrain. En el segundo lote, se comparó la descendencia heterocigota (Nn) de un macho Pietrain positivo al halotano (nn) con la descendencia libre del gen (NN) de dos machos Pietrain también libres del gen. Las hembras utilizadas en ambos lotes fueron las mismas, todas ellas libres del gen halotano

Suplementación de tranquilizantes naturales en la dieta de cerdos con diferente genotipo al estrés: sobre el bienestar animal y la calidad tecnológica y sensorial de la carne

El objetivo principal del proyecto era estudiar el efecto de tranquilizantes naturales sobre el bienestar de los animales y la calidad de la carne desde un punto de vista tecnológico y sensorial. Este objetivo se desglosaba en dos: OBJETIVO 1 – Evaluar la eficacia de tranquilizantes naturales (Magnesio, triptófano) en la disminución del nivel de estrés en cerdos de tres genotipos diferentes respecto del gen Hal, homocigotos dominantes NN, o portadores (Nn y nn) en el período anterior al sacrificio. OBJETIVO 2 – Estudiar el efecto de los tranquilizantes en la calidad tecnológica y sensorial de la carne (pH, capacidad de retención de agua, color, textura, olor sexual, …). Para ello se planteaban dos experimentos, el primero utilizando animales extremos, libres del gen, NN, y Hal +, nn, y el segundo con animales libres, NN, y portadores, Nn.

Valoración del bienestar animal del cerdo. Parámetros evaluados en el matadero

La preocupación por el bienestar animal es creciente en toda Europa, tanto por parte del consumidor como por parte del poder legislativo. La información que se facilite a los consumidores sobre los sistemas de producción y un etiquetado adecuado de las condiciones de producción de los productos de origen animal pueden constituir dos elementos esenciales de la cadena alimentaria europea en los próximos años. Es en este contexto que la Comisión Europea financia desde el año 2004 un proyecto europeo integrado, conocido con el acrónimo de “Welfare Quality®”. Uno de los objetivos principales del proyecto es obtener un sistema de valoración del bienestar de los animales de abasto que sea aplicable en granjas y mataderos, y que pueda convertirse en un sistema estandarizado para toda Europa. Además, el sistema de valoración, debe proporcionar información sobre el bienestar de los animales de una forma sencilla y entendible por el público y, a su vez, identificar de forma inequívoca los productos procedentes de estos estándares de bienestar animal. El sistema de valoración utilizado priorizará las medidas basadas en los propios animales sobre aquellas basadas en el ambiente o el manejo de estos. La inclusión de parámetros en el protocolo final de valoración del bienestar del porcino en el matadero depende de factores tales como su validez como indicador del bienestar animal, la facilidad con la que pueda ser valorado por distintos observadores en distintos entornos y condiciones con un error mínimo, que precise de poco tiempo para ser valorada y que combinada con otras medidas dé un resultado final óptimo. A modo de ejemplo, se presentan algunos de los parámetros que serán incluidos en el sistema de valoración del bienestar del porcino en matadero, tales como la valoración del miedo, resbalones, caídas, cojeras, presencia de heridas o índice de mortalidad y eficiencia del aturdimiento previo al sacrificio. Para más información se puede consultar la dirección www.welfarequality.net.

Novetats que afecten a la nutrició animal a Europa. Nou model europeu de producció animal

La nutrició animal a la Unió Europea (UE) ha estat afectada per vàries crisis, com la de les vaques boges (encefalopatia bovina espongiforme) (BSE), les hormones a la carn, la contaminació per les dioxines, els Organismes Genèticament Modificats (GMO) i l’ús d’antibiòtics com a promotors de creixement (AMGP). Des dels anys 70, la producció animal a la UE ha estat encaminada a buscar regulacions legals per a millorar la seva seguretat i eficàcia. Si més no, durant els últims cinc anys, casos com la BSE i d’altres han col·locat al sector en primera línia de les notícies, la qual cosa ha trencat la confiança dels consumidors en el consum de carn. Aquesta presentació intentarà explicar per què i com la UE està tractant aquests temes i què és el que el sector està fent per recobrar la confiança dels consumidors. La intenció és respondre a algunes qüestions que preocupen al sector, com per exemple: La producció animal europea, és menys segura que les altres? Per què les regulacions legals són més exigents per a nutrició animal que per als humans? El sector s’està preguntant si aquesta crisi pot produir un nou model europeu de producció animal. Finalment, l’actual sistema de ramaderia s’haurà de discutir: quantitat o qualitat.