Detecção de resíduos de enrofloxacina por ensaio imunoenzimático e cromatografia líquida acoplada à espectrometria de massas em ovos comerciais de galinha após tratamento / Investigation of enrofloxacin residues through enzyme-linked immunosorbent assay and liquid cromatography-tandem mass spectrometry in eggs from laying hens after treatment

A enrofloxacina é um dos antimicrobianos mais utilizados na avicultura industrial, e a deposição de resíduos em produtos avícolas, como os ovos, são de grande importância para a saúde pública. Na legislação brasileira não existe padronização do período de carência para o seu uso na produção avícola e não há Limite Máximo de Resíduo (LMR) fixado para enrofloxacina em ovos. Neste estudo, foi utilizado o kit de ELISA comercial (Bioo Scientific(r)) e a LC-MS/MS na pesquisa de enrofloxacina em ovos de 30 galinhas tratadas previamente via água de bebida, com 10mg/kg de enrofloxacina, durante cinco dias. Seis ovos foram coletados diariamente e analisados durante o tratamento e após a sua suspensão, durante 15 dias. A deposição de resíduos obteve níveis máximos no quinto dia de tratamento das aves, declinando gradativamente até não ser detectada a partir do nono dia de suspensão do tratamento. Considerando como base o LMR de 100µg/kg fixado pelo Brasil para tecidos comestíveis de aves e pela União Europeia para músculo, gordura e pele, após seis dias de suspensão do tratamento, os níveis de resíduos foram inferiores a esse limite, tendo como médias 37,43µg/kg na LC-MS/MS e 14,731µg/kg no ELISA. Dentro das condições deste estudo, um período de carência de seis dias seria mais adequado para utilização dos ovos para consumo humano. Foram detectados valores de resíduos nos ovos menores no ELISA em relação à LC-MS/MS para a mesma amostra, mas os dois métodos apresentaram concordância estatística entre si. A LC-MS/MS é o teste recomendado pela legislação brasileira para a análise de resíduos em alimentos; entretanto, pelos resultados obtidos, o kit de ELISA utilizado também pode ser aplicado na detecção de resíduos de enrofloxacina em ovos, com as vantagens de rapidez e simplicidade...

Enrofloxacin is one of the most used antibiotics in the poultry industry and the deposition of residues in poultry products, such as eggs, are of great concern to public health. In Brazilian law there is no standard withdrawal period for enrofloxacin in eggs and there is no Maximum Residue Limit (MRL) established for this antimicrobial in eggs. In this study, (Bioo Scientific(r)) commercial ELISA kit and LC-MS/MS were used to investigate enrofloxacin in eggs of 30 hens pretreated via drinking water at 10mg/kg of enrofloxacin for five days. Six eggs were collected daily and analyzed during treatment and after the end of treatment, for 15 days. Residues obtained maximum levels on the fifth day of treatment, declined gradually and were no longer detected from the ninth day to the end of treatment. Based on the MRL of 100mg/kg established for edible tissues of poultry by Brazillian law and for muscle, fat and skin, by the European Union, after six days of treatment withdrawal, the residue levels were below that limit, with the average of 37.43mg/kg in LC-MS/MS and 14.731mg/kg in ELISA. Within the conditions of this study, a withdrawal period of six days would be more appropriate to use the eggs for human consumption. The values obtained by ELISA for residues in eggs were lower than those obtained in LC-MS/MS for the same sample, however both methods showed statistical agreement. LC-MS/MS is the recommended method by Brazilian legislation for analysis of residues in food, however, according to the results the ELISA kit used can also be applied to the detection of enrofloxacin residues in eggs, with the advantages of speed and simplicity...

A method for the determination of total and reduced methimazole in various biological samples.

A simple, rapid, reproducible and sensitive method based on HPLC with ultraviolet detection was developed for the determination of methimazole (MMI) in animal tissues and plasma samples. Under the optimum experimental conditions, the calibration curves for MMI were linear in the tested range 0.5-20 mg kg(-1) tissue sample (mg l(-1) plasma) with correlation coefficients better than 0.99. The performance of the proposed method was tested for the determination of MMI levels in brain, liver, thyroid gland and plasma of MMI-treated hens, as well as in their eggs and embryos. The proposed method reduces time and simplifies the sample preparation procedure.

Surveillance of dioxins and polychlorinated biphenyls (PCBs) in food commercialized in Barcelona, Spain.

This study explored the potential use of seven congeners of polychlorinated biphenyls (PCBs-7) as indicator compounds for the presence of dioxins and PCBs in food samples, as part of the routine surveillance programme of a public health agency. Samples of 24 foodstuffs with high fat content were collected (ten fresh fish, six dairy products, five meat and three eggs). Duplicate analyses were performed. A research laboratory tested samples for seven polychlorinated dibenzo-p-dioxin (PCDDs), ten dibenzofurans (PCDFs) and twelve dioxin-like PCBs, with limits of detection in the range of ng kg(-1) (ppt). The public health services official control laboratory tested samples for PCBs-7, with a limit of quantification of 5 µg kg(-1) (ppb). The research laboratory detected the presence of dioxins and dioxin-like PCBs in all samples; fish samples had the highest levels (0.04-10.3 pg WHO-TEQ g(-1)). The public health service official control laboratory detected PCBs-7 only in five samples, which were all fish. Comparing the results in the two laboratories there seems to be an association between the detection of PCB-7 and the presence of higher levels of PCDD/Fs and dioxin-like PCBs. The use of PCB-7 as an indicator compound may be a practical surveillance strategy for those foodstuffs with higher concentrations of dioxin-like congeners.

Polybrominated diphenylethers (PBDEs) and brominated dioxins (PBDD/Fs) in Irish food of animal origin.

Brominated dioxins (PBDD/Fs) and 17 polybrominated diphenylether (PBDE) congeners including BDE-209 were analysed in 100 composite samples of commonly consumed foods of animal origin, prepared from ten to 40 individual subsamples of each type. These included 30 samples of milk from farms; eggs from chickens; fat from cows, pigs, sheep, and poultry; and liver from cows, pigs, sheep, horses, and poultry. The ISO 17025-accredited analytical methodology used, follows similar guidelines as given for chlorinated dioxins and polychlorinated biphenyls (PCBs) in European Commission Directive 2002/69/EC. Apart from some samples of milk, PBDD/Fs and PBDEs were detected in all food types. Concentrations ranged from 0.31 µg kg(-1) Σ17 PBDEs for a sample of milk to 37.5 µg kg(-1) Σ17 PBDEs for a sample of liver. The corresponding range for the PBDD/F toxicity (computed using analogous PCDD/F toxic equivalency factors (TEFs)) was 0.09 ng TEQ kg(-1) to approximately 3.5 ng TEQ kg(-1). The detection of these compounds, in particular PBDD/Fs, is consistent with the increased use and disposal of brominated organic compounds.

Organochlorine contaminants in cormorant, darter, egret, and ibis eggs from South Africa.

During the last 15 years, no research has been published on the levels of pesticides in bird eggs from South Africa, despite the high levels found previously. We analysed eggs from African darter, cattle egret, reed cormorant, African sacred ibis, as well as single eggs from some other species, and found HCB, DDTs, HCHs, chlordanes and PCBs at detectable levels. The presence of mirex in all species was unexpected, since this compound was never registered in South Africa. It also seemed as if terrestrial feeding birds had higher DDE:PCB ratios when compared with aquatic feeding birds. Except for chlordane, the African darter eggs had the highest levels of all other compounds (mean 370 and 300 ng g(-1) ww Sigma pesticides and Sigma PCBs, respectively). Multivariate analysis clearly distinguished the aquatic and terrestrially feeding birds on pollution profile. The African darter (aquatic feeding) and the cattle egret (terrestrial feeding) would be good indicator candidates. Eggshell thinning was detected in the African darter, and was associated with most of the compounds, including DDE and PCBs. We raise a concern that generally longer living birds in warmer climates, laying fewer eggs per clutch, might be at increased risk when compared with trophically similar birds exposed to equivalent levels of pollution in colder climates. Given the scarcity of water and the high biodiversity in Southern Africa, climate change will exert strong pressure, and any additional anthropogenic contamination at levels that can cause subtle behavioural, developmental and reproductive changes, can have serious effects.

Aquatic invertebrate resting egg sensitivity to glutaraldehyde and sodium hypochlorite.

Ballast tank treatment technologies are currently in development to reduce the risk of acquiring or transporting viable aquatic organisms that could be introduced to ecosystems and become invasive. Aquatic invertebrate resting eggs represent a challenge to such technologies because of morphological and biochemical adaptations to stress that also protect eggs from artificial stressors. To evaluate the potential efficacy of chemical biocides for ballast tank treatment, the present study examined the acute toxicity of glutaraldehyde and sodium hypochlorite on resting eggs of the freshwater cladoceran Daphnia mendotae and marine brine shrimp (Artemia sp.). Glutaraldehyde was toxic to resting eggs of Artemia sp., as indicated by a lethal concentration to 90% of organisms (LC90) of 95% confidence interval (226 +/- 10 mg/L). Daphnia mendotae, in contrast, displayed erratic responses to glutaraldehyde. Sodium hypochlorite was similarly toxic to resting eggs of Artemia sp. and D. mendotae, which displayed LC90s of 86.5 +/- 3.0 and 78.3 +/- 1.6 mg/L, respectively. Burial in sediment protected resting eggs from toxicants. The present results corroborate those from previous investigations of resting egg sensitivity to artificial stressors, supporting the conclusions that resting eggs are less sensitive than other life stages to artificial stressors and that chemical biocide concentrations effective against other life stages may be ineffective against resting stages.

Accumulation and transport behaviour of 241americium, 60cobalt and 134cesium by eggs of the spotted dogfish Scyliorhinus canicula.

An experimental study examined the 96-h net influx from seawater of the anthropogenic radionuclides (241)Am, (60)Co and (134)Cs through the egg-case of the spotted dogfish Scyliorhinus canicula. Net influx directly through the wall of the egg-case was greatest for (134)Cs, then (241)Am and lastly (60)Co. Within the egg-case wall itself the measured concentration factors (CFs) and their gradients in the external, median and internal layers showed that for both (241)Am and (60)Co they were >10(3) in the external layer and declined by an order of magnitude in the interior layer. In contrast (134)Cs had a CF of only about three in the external layer which declined by a factor of 2 towards the two more internal layers of the egg-case. The egg-case apertures, that open within the prehatching stage of embryological development, significantly (P<0.05) increased the net influx of (241)Am and (60)Co to the interior of the egg-case, although their water concentrations were still lower than those in the labelled seawater bath. In contrast, the aperture did not increase the net influx of (134)Cs whose water concentrations equilibrated with those in seawater. Together these results indicate that the egg-case wall is very permeable to (134)Cs, representing little barrier to its movement, and hence consistent with the lack of importance of the aperture in determining its internal water concentrations in the egg-case. In contrast, (241)Am and (60)Co show much higher rates of accumulation by the egg-case, consistent with the measured reduced permeability of its wall, and therefore giving greater prominence to its aperture in the net transfer of these two radionuclides to the egg case's interior. The presence of the embryo within its egg-case did not significantly (P>0.05) affect the rates of influx of radioisotopes, with the exception of an interactive effect for (60)Co with the egg-case aperture (P<0.05). The CF of only (241)Am in the embryo itself relative to the external seawater concentration was significantly (P<0.05) enhanced by the presence of the aperture.

Proliferative responses towards native, heat-denatured and pepsin-treated ovalbumin by peripheral blood mononuclear cells from patients with hen's egg-sensitive atopic dermatitis.

In order to clarify the mechanism of food-antigen recognition, the proliferative responses of peripheral blood mononuclear cells (PBMCs) to native, heat-denatured or pepsin-treated ovalbumin (OA) were investigated in 16 hen's egg-sensitive patients with atopic dermatitis (AD). Seven of them had hypersensitivity to boiled hen's egg and others had not. The responses of PBMCs to heat-denatured OA were lower than those to native OA in the patients without hypersensitivity to boiled hen's egg. However, there were no differences of the responses of PBMCs between heat-denatured OA and native OA in the patients with hypersensitivity to boiled hen's egg. Moreover, the reduction of the responses of PBMCs to pepsin-treated OA was recognized in six out of seven patients. The primary structure of OA did not change by heating or pepsin treatment according to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). These results suggested that the secondary structure of OA changed in connection with the reduction of the responses of PBMCs to denatured OA. In addition, we demonstrated the suppressive effect of anti-HLA-DR and anti-HLA-DQ monoclonal antibodies on the proliferative response of PBMCs to OA. The results suggested that the proliferative responses of PBMCs to OA were restricted by HLA-DR or HLA-DQ in hen's egg-sensitive patients with AD.

Epidemiology & Disease Control Program. Update: Salmonella enteritidis associated with shell eggs. February, 1992.

In previous articles in the Maryland Medical Journal we have reviewed the problem of Salmonella enteritidis associated with shell eggs. Cases and outbreaks continue to occur. Health care professionals need to be aware of the association between the consumption of raw or undercooked eggs or egg-containing foods and gastroenteritis, especially in high risk individuals.