Foodstuffs Of Animal Origin Research Articles

Antibiotics in food: Legislation and validation of analytical methodologies

In this paper, European food safety legislation is presented, and special attention is devoted to monitoring residues of veterinary drugs in foodstuffs of animal origin. After a short review of the state of the art of analytical methodology for antibiotic residue analysis, the paper focuses on validation of analytical methods, with Decision 657/2002/EC as reference document. Finally, the main issues of the quality control of the analytical data, i.e. analysis of reference materials and participation in proficiency tests, are briefly addressed.

Evaluation of 10-years French NRL proficiency tests for lead, cadmium and mercury analysis in foodstuff of animal origin

The French National Reference Laboratory (NRL) for heavy metals in foodstuffs of animal origin at the French Food Safety Agency (AFSSA) regularly organises proficiency tests schemes (PTS) focused on the quantification of the three elements lead (Pb), cadmium (Cd) and mercury (Hg), for the 23 departmental laboratories of the network of laboratories, under its responsibility. The NRL prepares test materials to promote comparability of data and support harmonisation of official analytical procedures in the case of routine analysis. When target values for the analytes are set, the PTS also provide an objective assessment and improvement of the analytical trueness in the daily work of the laboratories. The long-term performance of the network was examined using results of 10 years of PTS in various matrices from different foodstuffs of animal origin (fish, meat and milk products), with 3 different official methods, using Flame- and Electro-Thermal-AAS techniques. In all cases, participants were requested to carry out 3 independent measurements of Pb, Cd and Hg. Moreover, the assessment of the PTS was based on the Z-score approach. From 1998 to 2008, this evaluation applied to 690 results obtained in 41 PTS indicated a frequency distribution of the Z-score value for the three elements Pb, Cd and Hg of 89.2%, 94.5% and 97.7% of acceptable results, respectively. The results of this balance sheet indicated the homogeneity and ability of the laboratories network to provide comparable results of similar quality. Comparison with results obtained by a primary method were presented as complementary information to determine the interest to organize, in the future, PTS based on a certified value.

Main PaperInnovative swine nutrition: some present and potential applications of latest scientific findings for safe pork production

One of the biggest challenges facing animal agriculture in the 21st century is to produce safe and traceable foodstuffs of animal origin in sufficient volumes and quality besides the lowest possible load on the environment. A point to consider is how animal feeding and nutrition can contribute to the solution of this challenge. Another question is, that the results of which new scientific fields should be used in the area of innovation to achieve the desired product quality. The article answers these questions based on the latest data in the scientific literature and on the authors’ own research results. Having reviewed and processed the data, the authors drew the following conclusions:1. The impact of climate changes on crop production and consequently on animal nutrition needs to be studied in comprehensive, systematic research programs, and based on their results animal feeding systems should be modified if necessary.2. The role of molecular nutrition and the immunological role of nutrition for enhancing the efficiency of production will gain in importance.3. The cooperation between nutritionists and geneticists needs to be strengthened in order to satisfy the nutrient requirements more accurately, and thus to improve the profitability of production.4. New mathematical growth models, also incorporating the quality of animal products, need to be developed for better production estimates.5. The integrated “from farm to fork” programs will be essential in the production of safe and high quality animal food products. Animal nutrition will play a key role in these product development and monitoring programs.6. The production of environmentally friendly animal products will become a social imperative. Animal nutrition still has huge unexploited reserves in this field, for instance in the development of novel animal feeding systems and technologies.7. The practical implementation of precision nutrition can be of great help in achieving these goals and in improving the efficiency innovation activities.

Hiérarchiser les risques de zoonoses alimentaires : une aproche quantitative.

Foodborne zoonoses are a major public health concern. Risk analysis, which underpins international policies on food safety and trade in foodstuffs of animal origin, requires that an assessment be made of the occurrence and severity of human cases for each type of foodstuff. However, the tools currently available for quantifying risks are only capable of estimating the consequences of certain diseases. This article proposes an alternative quantitative approach for prioritising the risk of foodborne zoonoses, based on the creation of a typology of hazards and calculating a risk score. A combination of average hospitalisation and mortality rates is used to quantify the severity of human cases. By calculating the percentage of food-associated cases it is possible to estimate the incidence of cases linked specifically with the foodstuff being assessed. This method is illustrated by applying it to bacterial zoonotic hazards in pork and beef and provides a support tool for veterinary public health decision-makers.

High-throughput multiclass method for antibiotic residue analysis by liquid chromatography–tandem mass spectrometry

A simple and rapid method has been developed for the residue analysis of 39 antibiotics (tetracyclines, quinolones, penicillins, sulfonamides and macrolides) in foodstuffs of animal origin. The method combines an effective extraction technique, which uses water–methanol as extracting solvent, with ultra-high-pressure liquid chromatography–tandem mass spectrometry, allowing both confirmation and quantification in a single chromatographic run. The multiresidue method has been validated in chicken muscle matrix according to European Union Decision 2002/657/EC. It has been implemented as a routine method in a Public Health Laboratory, instead of the five plates test and LC methods previously used.

Chromatographic Residue Analysis of Sulfonamides in Foodstuffs of Animal Origin

The widespread use of antibiotics in veterinary medicine for prophylactic, therapeutic and growth promoting purposes may result in the presence of residues in foodstuff of animal origin. Sulfonamides, N‐derivatives of 4‐amino‐benzenesulfonamide, belong to a diverse class of synthetic antibiotics consisting of chemically related compounds. A number of them e.g., sulfamethazine, sulfadiazine, sulfaquinoxaline and sulfamethoxypyridazine, are widely used in animal husbandry. Their residues in food are of great concern because of their potential carcinogenic character and because they may contribute to the development of antibiotic resistance in humans. These facts have induced most of the regulatory activities on sulfonamides. Regulatory bodies in the European Union as well as in other countries have set different maximum residue limits in various tissues. These regulations have stimulated the development of fast, accurate and sensitive analytical methods for monitoring sulfonamide residues in food samples of animal origin. Gas chromatography methods tend to be time‐consuming due to the required previous cleanup and derivatization procedures, thus, HPLC methods with UV or MS detection have the leading position. These and other aspects of current analytical methodology, including sample preparation and cleanup, are reviewed and discussed. Emphasis is given to confirmatory methods since these comprise useful tools to regulatory agencies, and identifications based on these methods can be used in support of regulatory action.

Optimisation and critical evaluation of a collision cell technology ICP-MS system for the determination of arsenic in foodstuffs of animal origin

The determination of arsenic ( 75As) was studied using an ICP-MS equipped with collision cell technology (CCT). Different mixtures of gases (He and H 2) were tested using HCl conditions and a He flow rate of 4 mL min −1 was found to be suitable for the removal of the poly-atomic spectral interference [ 40Ar 35Cl] +. Trueness of the optimised method has been evaluated in both standard and CCT modes on six certified reference materials in foodstuffs of animal origin and on three external proficiency testing schemes (FAPAS). The results obtained generally coincided with the certified values, except for CCT mode in some categories of samples (meat, mussels and milk powder), for which a positive bias on results was observed due to the formation of poly-atomic interferences within the collision cell. The main interferences were studied and their contributions estimated. [ 58Fe 16O 1H] + and [ 74Ge 1H] + were the most significant interferences formed in the cell. Finally, different parameters (e.g. hexapole and quadrupole bias voltage, nebuliser gas flow) were optimised to try to attenuate these interferences.

Validation of two enzyme immunoassays for aminoglycoside residues according to European Decision 657/2002

Aminoglycoside antibiotics are commonly used to treat bacterial infections in human and veterinary practice. Owing to their toxicity, the European Community has established maximum residue limits (MRL) in foodstuffs of animal origin (EEC No 2377/90). In the present study, the performance of two new enzyme immunoassays (EIA), I'screen Gentamicin and I'screen Neomycin, for the quantitative detection of the aminoglycosides, gentamicin and neomycin, in milk and tissue are described. Validation of these EIAs has been performed in accordance to the criteria of European Decision 657/2002. Assays sensitivity at the MRLs was 95% for milk samples and 100% for tissue samples, while specificity was 100% at 33 and 25% of the MRLs for milk and tissues, respectively. The performance of these EIAs indicates that they can be used as easy screening methods for the analysis of aminoglycosides in milk and tissue samples.

Analytical methods for multiresidue determination of sulfonamides and trimethoprim in meat and ground water samples by CE‐MS and CE‐MS/MS

This paper presents two methods based on CZE-MS detection and CZE-MS/MS detection developed for the multiresidue determination of ten sulfonamides (sulfapyridine, sulfadoxin, sulfamethazine, sulfadimethoxine, sulfameter, sulfamerazine, sulfachlorpyridazine, sulfadiazine, sulfamethoxazole, and sulfamethizole) and a potentiator, trimethoprim (TMP), whose contents are regulated by the EU Council Regulation no. 2377/90 in animal edible tissues. Experimental designs were employed to optimize the electrospray conditions. MS/MS experiments using an IT as analyzer operating in multiple reaction monitoring (MRM) mode were carried out to achieve the minimum number of points according to the 2002/657/EC European Decision for unambiguous identification. The proposed procedures have been compared in terms of the performance characteristics and trueness. The limits of detection and quantification were in all cases lower than the maximum residue limits legislated for these compounds and the recoveries were satisfactory, being possible the application for their monitoring in foodstuff of animal origin and in environmental samples, allowing the determination of sulfonamides and TMP residues in meat and in superficial water in the low microg/L range.

Investigation of palladium and platinum levels in food by sector field inductively coupled plasma mass spectrometry

Over the last two decades, there has been increased concern regarding the impact of some noble metals, such as Pd and Pt, on human health. These elements pollute the environment due to their widespread use as catalytic converters and in medical applications. The risk they pose to human health and the environment is still controversial; however, literature data point to diet as an important source of uptake by the human body. Within this context, the total Pd and Pt content of several Italian food commodities has been investigated. A total of 90 samples, including flour products, vegetables and foodstuffs of animal origin (meat, milk and eggs), were collected and freeze-dried. Samples were analyzed by sector field inductively coupled plasma mass spectrometry (SF-ICP-MS) after chopping or crushing followed by freeze-drying and microwave (MW)-assisted acid digestion in a Class-100 clean-room. A mathematical approach was adopted to correct the mass signals for still unresolved interference (mΔm = 300, 10 000). The lowest and highest concentrations of Pt, i.e. 17 and 93 ng kg−1 (dry weight, dw), were found in vegetables and flour products, respectively. The lowest Pd level (2830 ng kg−1 dw) was found in eggs and the highest (47 800 ng kg−1 dw) in vegetables.

Advances in Chromatographic Analysis of Tetracyclines in Foodstuffs of Animal Origin—A Review

Tetracyclines (TCs) are broad‐spectrum antibiotics in human and veterinary medicine, characterized by their exceptional chemotherapeutic efficacy against a wide range of Gram‐positive and Gram‐negative bacteria, rickettsiae, spirochetes, large viruses, chlamydia, mycoplasmas and protozoan parasites. The use of tetracyclines is increasing, as they are used not only in treatment but also prevention of illnesses. Moreover TCs are given to animals destined for human consumption to promote growth and may potentially result in the presence of residues in edible animal tissues, which can be toxic and dangerous for human health and potentially cause allergic reactions. An extended and comprehensive review on the analytical methodologies concerning tetracyclines in foodstuffs of animal origin reported in literature is provided in the present article. Emphasis is given on sample preparation regarding isolation and purification, chromatographic conditions and method validation according to the legislation. Results of published assays are comparatively presented and criticized.

Residues of certain veterinary drugs in raw milk in Slovenia in the 2000 2002 period

In order to ensure the hygienic suitability of foodstuffs of animal origin, and from the aspect of human health protection and improvement, we studied the contamination of raw milk with microbial inhibitors, chloramphenicol, sulphonamides and ivermectin in one part of Slovenia - the central Stajerska region. Our own sampling in 2002 included industrial areas along with areas of intensive and extensive milk production. Thus, in the summer, autumn and winter periods of that year, 27 samples of raw milk were taken and tested for residues of microbial inhibitors, chloramphenicol, sulphonamides and ivermectin. Microbial inhibitors, sulphonamides and ivermectin were, in fact, not found in any of the tested samples. Yet in two samples of raw milk taken in the autumn of 2002, the presence of chloramphenicol was confirmed. These results of testing milk from the central Stajerska region were compared with the results of systematic veterinary-sanitary control in Slovenia during 2000 and 2001 (the residual monitoring programme). No residues of certain veterinary drugs tested for were found in any of the samples that were examined.

Food-Borne Zoonoses and the Consumer

The aim was to analyze the role of fear and panic caused by a zoonosis in the development of eating habits and rituals. The news media, being the source of information for the consumers about infectious agents having a high morbidity and mortality, can cause fear and panic about foodstuffs of animal origin. In this way, they play an important role during changes in eating habits, many of which become rituals. Rituals affect consumers' behavior and perceived needs regardless of socio-economic class, thus they increase consumption. Hence, the mass news media for selling profit-making information or promoting one kind of food against another, or the products of one country against those of another could use fear and panic caused by zoonoses.

Determination of chromium, iron and selenium in foodstuffs of animal origin by collision cell technology, inductively coupled plasma mass spectrometry (ICP-MS), after closed vessel microwave digestion

The determination of chromium ( 52Cr), iron ( 56Fe) and selenium ( 80Se) isotopes in foodstuffs of animal origin has been performed by collision cell technology (CCT) mode using an inductively coupled plasma mass spectrometry (ICP-MS) as detector after closed vessel microwave digestion. To significantly decrease the argon-based interferences at mass to charge ratios ( m/ z): 52 ( 40Ar 12C), 56 ( 40Ar 16O) and 80 ( 40Ar 40Ar), the gas-flow rates of a helium and hydrogen mixture used in the hexapole collision cell were optimised to 1.5 ml min −1 H 2 and 0.5 ml min −1 He and the quadrupole bias was adjusted daily between −2 and −15 mV. Limits of quantification (LOQ) of 0.025, 0.086 and 0.041 mg kg −1 for Cr, Fe and Se, respectively, in 6% HNO 3 were estimated under optimized CCT conditions. These LOQ were improved by a factor of approximately 10 for each element compared to standard mode. Precision under repeatability, intermediate precision reproducibility and trueness have been tested on nine different certified reference materials in foodstuffs of animal origin and on an external proficiency testing scheme. The results obtained for chromium, iron and selenium were in all cases in good agreement with the certified values and trueness was improved, compared to those obtained in standard mode.

Ochratoxin A in human blood in Abidjan, Côte d'Ivoire

Ochratoxin A (OTA) produced by Aspergillus and Penicillium genera contaminates a diversity of foods including cereals; cereals-derived foods; dry fruits; beans; cocoa; coffee; beer; wine; and foodstuffs of animal origin mainly poultry, eggs, pork and milk, including human breast milk. OTA is nephrotoxic to all animal species studied so far and most likely to humans, who show the longest half-life for elimination of this toxin among all species examined. Among other toxic effects, OTA is teratogenic, immunotoxic, genotoxic, mutagenic and carcinogenic, all of which lead to life-threatening pathologies through several molecular pathways. In Côte d'Ivoire, preliminary surveys conducted by us have proven from 1998 to 2004 the reality of ochratoxin A-contamination of foodstuffs. To assess OTA in human blood, the immunoaffinity columns were used along with HPLC for separation and fluorimetric quantification of blood samples collected in Abidjan from two categories of people: apparently healthy donors ( n=63) and nephropathy patients undergoing dialysis ( n=39). Among healthy donors, 34.9% show OTA concentrations ranging from 0.01 – 5.81 μg/l with a mean value of 0.83 μg/l, whereas, among nephropathy patients undergoing dialysis 20.5% are OTA positive in a range of 0.167–2.42 μg/l and a mean value of 1.05. Although the sex ratio is 0.82 (46 females for 56 males) ochratoxin A contamination is equally distributed in both sexes. Nephropathy patients undergoing dialysis appear, however, less frequently contaminated than healthy donors (20.5 versus 34.9%) and show higher OTA concentrations (higher mean value, p=0.01). Ochratoxin A concentrations found in human blood reflect concentrations previously detected in cereals and peanuts according to the eating habits and diets of people in Côte d'Ivoire. But, the prevalence of ochratoxin A in blood of nephropathy people undergoing dialysis appears lower than expected from the frequency of OTA contamination in cereals and peanuts. Pearson χ 2-test indicates that among OTA-positive individuals renal dialysis and age are important modalities for consideration.

Determination of β-lactam residues in foodstuffs of animal origin using supported liquid membrane extraction and liquid chromatography–mass spectrometry

β-Lactams have been used extensively both in human and veterinary medicine practices. In veterinary medicine, they are used mainly as growth promoters as well as chemotherapeutic and prophylactic agents. The occurrence of β-lactam residues in foodstuffs is a serious health hazard. In this work, a sample purification and enrichment technique involving the use of supported liquid membrane have been developed for β-lactams in milk, kidney and liver tissues. The liquid membrane made of n-undecane:di- n-hexyl ether (1:1) was used to enrich a mixture of four β-lactams namely, ampicillin, cloxacillin, penicillin V and penicillin G. Separation and detection of the enriched β-lactam extracts were performed using a high performance liquid chromatography coupled to a mass spectrometer operating under positive ion electrospray mode (LC–PI–ESI–MS). The detection limits (DLs) obtained were found to be 1 ng/kg for penicillin G and penicillin V in kidney and liver tissues and 0.7 μg/L in milk. For ampicillin, the DLs were found to be 1.4 μg/kg in kidney and liver tissues and 1.7 μg/L in milk. Limits of quantification based on a minimal value of the signal-to-noise ratio of 10, were estimated to be 1.1 and 0.01 for penicillin G in kidney and liver tissues, respectively, while for penicillin V it was 0.4 and 0.01 for kidney and liver tissues, respectively. The DL values obtained using this approach were found to be 2–3 order of magnitudes lower than the stipulated tolerance levels as set by the EU and FDA.

Simultaneous Analysis of Cadmium, Lead, Mercury, and Arsenic Content in Foodstuffs of Animal Origin by Inductively Coupled Plasma/Mass Spectrometry after Closed Vessel Microwave Digestion: Method Validation

A method validation of the total analysis of cadmium (Cd), lead (Pb), arsenic (As), and mercury (Hg) in foodstuffs by inductively coupled plasma/mass spectrometry (ICP/MS) after closed vessel microwave digestion is presented. Due to the lack of reference method for ICP/MS techniques in food and, based on the project of the European Committee of Normalization (CEN/TC 275/WG 10), the Agence Française de Normalisation (AFNOR) guidelines NF V03-110 were used for the evaluation of this method based on 2 steps, sample preparation and multielement detection. Several criteria considered as compulsory (linearity, specificity, precision under repeatability conditions, and trueness) have been estimated and discussed, in addition to intermediate precision reproducibility, the limit of detection, and the limit of quantification. Furthermore, a comparison with in-house methods using electrothermal atomic absorption was performed using an external proficiency testing scheme and food samples. The results indicated that this method could be used in the laboratory for the routine determination of these 4 cumulative toxic metals in foodstuffs with acceptable analytical performance.

Prevalence of Listeria monocytogenes in milk, meat and foodstuff of animal origin and the phenotype of antibiotic resistance of isolated strains

In 2000–2002 samples of raw materials (milk and beef, 518 samples), ready-to-cook foods and final products from milk (30 and 200 samples) and from meat (105 and 110 samples) and swabs from surfaces in two meat-processing plants (41 samples) were examined for the presence of Listeria monocytogenes (L. monocytogenes). 70 isolates were tested using the standard microdilution method for the susceptibility to 12 antimicrobial drugs, minimum inhibitory concentration (MIC) characteristics (MIC<sub>m</sub>, MIC<sub>50</sub>, MIC<sub>90</sub>) were determined. L. monocytogenes was isolated from raw milk samples (15 samples). It was not isolated from any of the semi-finished and final milk products except for one sample of pasteurized milk. Furthermore L. monocytogenes was isolated from samples of raw beef, meat emulsion, fermented dry meat products and from swabs from production equipment. In vitro testing of susceptibility showed the considerable effectiveness of examined antimicrobial substances with streptomycin and norfloxacin being least effective (MIC<sub>m</sub> = 4 μg/ml). Apart from the intermediate resistance (MIC = 1–2 μg/ml) and resistance (MIC = 4 μg/ml) to clindamycin (37 and 5 strains, respectively), all strains were susceptible to the tested substances. While the presence of L. monocytogenes in foodstuffs, in particular, are serious, current tests of susceptibility of L. monocytogenes isolates indicate low probability untreatable infections as a result of resistant strains from foods or transfer of resistance to other microorganisms in the CzechRepublic.

Prevention and control of mycotoxins in the poultry production chain: a European view

Prevention and control of mycotoxins in the poultry production chain requires the knowledge and consideration of all factors influencing mycotoxin formation in the field and during the storage of feedstuffs. Mycotoxin effects on health and performance of poultry and the “carry over” of mycotoxins or their metabolites into foodstuffs of animal origin are also essential information elements.Among a number of known mycotoxins, the Fusarium-mycotoxins deoxynivalenol (DON) and zearalenone (ZON) are of special importance under European production conditions because of their high frequency and concentrations. Several plant production strategies might help to minimize the risk of formation of these mycotoxins. However, higher mean values are unavoidable under unfavourable weather conditions. Therefore, special care must be taken in so-called “Fusarium-years”, because poultry is regarded as less sensitive to these mycotoxins than pigs and suspect cereal batches will be diverted to poultry feeding.Higher dietary DON-concentrations adversely affect the performance and immune response of poultry at dietary concentrations of greater than 5 mg per kg diet. In contrast, zearalenone, which often co-occurs with DON, does not cause deleterious effects in poultry even in “Fusarium-years” when higher concentrations are measured. “Carry over” of both toxins into poultry products is very low and is negligible when the DON-concentration of the diet is restricted to this 5 mg per kg.The levels of further Fusarium mycotoxins such as T-2 toxin, fumonisins, moniliformin or beauvericin to be expected in Europe are too low to cause toxic effects in poultry. However, several combinations of co-occurring mycotoxins might influence the total toxic potential of a contaminated feedstuff.Several feed-processing procedures are suitable for reducing the DON-content of contaminated feedstuffs, but they are expensive or have been only lab-tested. Adsorbing agents were shown to either only weakly bind DON, or not to bind it at all in vitro, and to be ineffective in vivo. More recent efforts have been directed at employing the capability of several micro-organisms (bacteria and yeasts) to degrade DON for detoxification in situ.

Determination of ciprofloxacin, enrofloxacin and flumequine in pig plasma samples by capillary isotachophoresis–capillary zone electrophoresis

Quinolones are a group of synthetic antibiotics that are widely used in veterinary medicine. Their residues may remain in tissues, milk, etc. intended for human consumption. The European Union fixes the maximum residue limits (MRLs) of veterinary medicinal products in foodstuffs of animal origin. Analytical methods are therefore needed to determine them in biological samples. In this study, we describe capillary isotachophoresis–capillary zone electrophoresis (ITP–CZE) to analyze three quinolones, enrofloxacin (ENR), ciprofloxacin (CPR) and flumequine (FLU), in pig plasma samples. We used solid-phase extraction with Oasis HLB cartridges as a sample pretreatment clean-up step. Capillary zone electrophoresis (CZE) requires low amounts of sample and is not as sensitive as one would wish. ITP–CZE is an easy way to increase the sample loadability and sensitivity. With this system sensitivity increases 40-fold. The detection limits for CPR, ENR and FLU were 70, 85 and 50 μg l −1, respectively, which were lower than their MRLs in different kinds of samples. This method is simple and sensitive, and is therefore an alternative tool to the existing HPLC methods for analyzing the residuals of these quinolones in biological samples.