Pig Feed Samples Research Articles

Validation of an Enzyme-Linked Immunosorbent Assay for Detecting Sulfonamides in Feed Resources

An enzyme-linked immunosorbent assay (ELISA) to screen sulfadiazine and sulfamethazine residues in feeds has been developed and validated according to Commission Decision 2002/657/EC criteria. Sulfonamides are easily extracted with a 95:5 acetonitrile/water mixture, obtaining recoveries between 80 and 100%. Accuracy, precision, selectivity, robustness, limit of detection (LOD), and detection capability (CCbeta) of the assay have been assessed during the validation process. LOD values in pig feed samples were 0.2 microg/g for sulfadiazine and 0.04 microg/g for sulfamethazine without any sample treatment other than extraction, dilution with the assay buffer, and filtering of the resulting solution. Furthermore, a new strategy for the determination of CCbeta in an ELISA screening method is proposed; this gave CCbeta values of 0.8 microg/g for sulfadiazine and 0.1 microg/g for sulfamethazine. Besides sulfadiazine and sulfamethazine, other sulfonamides can be detected with this immunoassay; this has been verified calculating their LOD values and cross-reactivities. Finally, real feed samples were analyzed with the ELISA methodology and a previously developed liquid chromatography (LC) method, and results confirmed the utility of this new immunoassay for screening purposes.

Lateral-flow colloidal gold-based immunoassay for the rapid detection of deoxynivalenol with two indicator ranges

A lateral-flow immunoassay using a colloidal gold-labelled monoclonal antibody was developed for the rapid detection of deoxynivalenol (DON). Different parameters, such as the amount of immunoreagents, type of the materials, composition of the blocking solution and of the detector reagent mixture, were investigated to provide the optimum assay performance. The experimental results demonstrated that such a visual test had an indicator range rather than a cut-off value. Thus, tests for DON determination with two different indicator ranges of 250–500 and 1000–2000 μg kg −1 were designed. The method allowed detection of DON at low and high concentration levels, which could be useful for research and practical purposes. The assay applied to spiked wheat and pig feed samples demonstrated accurate and reproducible results. The applicability of the developed lateral-flow test was also confirmed under real field conditions. The test strips prepared in Belgium were sent to Mexico, where they were used for the screening of DON contamination in different bread wheat entries from Fusarium Head Blight inoculated plots. The results were compared with those obtained by ELISA and LC–MS/MS. A poor correlation between ELISA and LC–MS/MS was observed. Visual results of the dipstick tests were in a good agreement with the results of the LC–MS/MS method. Coupled with a simple and fast sample preparation, this qualitative one-step test based on the visual evaluation of results did not require any equipment. Results could be obtained within 10 min. The described assay format can be used as a simple, rapid, cost-effective and robust on-site screening tool for mycotoxin contamination in different agricultural commodities.

Liquid chromatographic method development for anabolic androgenic steroids using a monolithic column: Application to animal feed samples

An isocratic HPLC method for the determination with screening purposes of anabolic androgenic steroids (AASs: fluoxymesterone, boldenone, nortestosterone, metandrostenolone, norethindrone, methyltestosterone and bolasterone), used as growth promoting agents, in finishing pig feed samples has been developed and validated. The separation was achieved by using a reversed-phase Chromolith RP-18e column at controlled temperature, UV-detection at 245 nm and epitestosterone as internal standard. The method development involved optimization of different aqueous-organic mobile phases using methanol or acetonitrile as organic modifiers, flow-rate and temperature. The optimum separation for these compounds was achieved at 40 °C using ultrapure water:acetonitrile (71:29, v/v) as mobile phase and 3 mL min −1 flow-rate, allowing the separation of AASs with baseline resolution in about 15 min. The optimized method was applied to the analysis of AASs in finishing pig feed samples. Prior to HPLC, sample preparation procedure was used by leaching using acetonitrile, saponification in a basic medium and solid-phase extraction using polymeric Abselut Nexus cartridges. Method validation has been carried out according to the European Commission Decision 2002/657/EC. The extraction efficiencies, decision limits (CCα) and detection capabilities (CCβ) for these compounds were in the range 83–96%, 27–37 and 32–47 μg kg −1 range, respectively. The within-laboratory reproducibility at 1, 1.5 and 2 CCβ concentration levels were smaller than 13, 10 and 8%, respectively. Finally, the proposed method was successfully applied to nine different kinds of animal feed.

Performance of pigs reared under traditional tribal low input production system and chemical composition of non-conventional tropical plants used as pig feed

Abstract The present study was carried out to document the pig rearing practices followed by the tribal farmers, to analyze the proximate composition of commonly used non-conventional feed stuffs and to study the growth performance of pigs under low input traditional pig production system. The study was conducted over a period of two and half years (2003–2005) in Mizoram, a hilly tribal state of North Eastern India. A total number of 320 farmers were surveyed all over the state (40 farmers per district in eight districts). The non-conventional pig feed materials/plants were collected, identified and evaluated for proximate composition. Besides this, a total number of 200 home made cooked ready to serve pig feed samples were collected from all over the state and analyzed for proximate composition. The average dry matter, crude protein, crude fiber, ether extract, total ash and nitrogen free extract content of ready to serve pig feed were 14.80 ± 2.79, 6.66 ± 2.14, 31.65 ± 8.09, 4.71 ± 1.99, 6.89 ± 2.27 and 50.17 ± 5.86, respectively. Eight male and seven female piglets of each Hampshire, Large White Yorkshire (LWY) and Nondescript local pigs, reared by 15 different farmers were selected randomly to assess the growth performance under low input production system. There was significant (p

Surveillance of Toxigenic Fungi and Ochratoxin A in Feedstuffs from Córdoba Province, Argentina

The aim of this work was to evaluate the incidence of potential ochratoxigenic mycoflora and ochratoxin A (OA) in poultry, pig and rabbit feeds. Eighty poultry, pig and rabbit feed samples were taken at random from factories located from Córdoba province, Argentina, over a period of 8 months. Isolation and quantitative enumeration of fungal propagules were done on DRBC and DG18 media. The predominant species were A. candidus, A. flavus, A. terreus, A. parasiticus, P. implicatum, P. minioluteum, P. crustosum and P. citrionigrum. The distribution of section Nigri species varied according to the feedstuffs analysed. The frequency of A. niger var. niger was noticeably high in poultry feed samples on DRBC medium. The Nigri section species was present at moderate mean colony counts (CFU/g) from three feeds. Mycotoxin analysis of these samples showed that OA was detected in 15%, 10% and 12% of pig, poultry and rabbit feed samples, respectively. The mean levels detected ranged between 15 and 25 ng/g from three feeds. The presence of ochratoxigenic species of Nigri section and OA in feeds indicates the risk of potential exposure of poultry, pigs and rabbits through the ingestion of feeds.

Concentrations of additive arsenic in Beijing pig feeds and the residues in pig manure

The swine industry in China has grown rapidly over last two decades. Great amount of pig manure is generated in China, which can be used as organic fertilizers on agricultural lands. Meanwhile, the organic arsenic compounds have been used as feed additives for swine disease control and weight improvement. Once the excessive additives are released in the environment, arsenic may compromise food safety and environmental quality. There is a growing public concern about the arsenic residues accumulation in pig manure, however, little work has been done to investigate the exact arsenic content in pig feed and the residues in manure in China This study investigates the concentrations of arsenic in 29 pig feed samples and 29 manure samples collected from eight pig farms in the Chaoyang district, Beijing city. The detected rate of arsenic in 29 couples of samples was 100%. The concentrations of arsenic in pig feeds and manures ranged from 0.15 to 37.8 mg/kg and 0.42 to 119.0 mg/kg, respectively. The result showed that arsenic concentration in pig manure will be greatly elevated when the arsenic in pig feed was largely increased. The loading rates of pig manure in fourteen Beijing counties and districts were in the range of 2.7–57.2 t/ha yr. Accordingly, the potential soil arsenic increase rates resulting from land application of pig manure might range between 11.8 and 78.9 μg/kg yr. Despite these findings, it is too early to draw the conclusion that arsenic pollution from pig manure is serious in Beijing farmland; therefore, longitudinal studies about the chemical form transformation and the environmental behaviors of pig manure arsenic are required in order to come up with more definitive conclusions.

Evaluation of Dry Ashing in Conjunction with Ion Chromatographic Determination of Transition Metal Ions in Pig Feed Samples

The contents of transition metal ions iron, copper, zinc, and manganese were simultaneously determined in pig feed using an ion chromatographic technique (IC) preceded by dry ashing. Employing ion exchange, the ions were separated on an IonPac CS5A column used in combination with a pyridine-2,6-dicarboxylic acid based eluent. The separation was followed by spectrophotometric detection after postcolumn reaction with 4-(2-pyridylazo)resorcinol. Dry ashing parameters were varied to assess their role in potential analyte loss. Quantitative recoveries (>95%) were obtained for all analytes with a dry ashing method that included a moderate temperature-time regime and ash leaching support in the form of sonication and heat treatment. The use of HCl as leaching acid and the presence of alkaline earths in the matrix solution did not interfere with the chromatographic separation.

Detection of ochratoxin A in animal feeds and capacity to produce this mycotoxin by Aspergillus section Nigri in Argentina

Ochratoxin A (OA) is a mycotoxin detected in a variety of food and feeds mostly from countries with a temperate climate because of the fungi that produce it, mainly Aspergillus ochraceus and Penicillium verrucosum. In Argentina, there is no available information about the natural occurrence of OA and ochratoxigenic fungi from feedstuffs. The aim was to evaluate the natural occurrence of OA in poultry, pig and rabbit feeds over 8 months. Likewise, the capacity to produce OA by Aspergillus section Nigri was investigated. Mycotoxin analysis showed that in some months of sampling, OA was detected in three feeds. OA was found in 38% of the poultry feed samples tested with levels ranging from 25 to 30 ng g−1. From rabbit feed samples, 25% contained OA and the levels ranged from 18.5 to 25 ng g−1. Only 13% of the pig feed samples were contaminated with similar levels of toxins. Ninety-four black Aspergillus strains from feedstuffs were tested for OA production. Among these, the tested species were A. niger var. niger, A. niger var. awamori, A. japonicus var. japonicus, A. japonicus var. aculeatus and A. foetidus. For the detection of OA, three methodologies were applied: the two TLC methods used for the fast screening of the filamentous fungi for the production of OA were not sensitive enough to detect OA in any of the black Aspergillus strains. When an HPLC methodology was used, the results showed that 46% of the black Aspergillus strains were producers of OA, with levels ranging from 13 to 25 ng ml−1 culture medium. The highest percentage of ochratoxicogenic strains was isolated from rabbit feeds with 100 and 78% of A. niger var. niger and A. niger var. awamori, with mean levels of 15.5 and 14.6 ng ml−1, respectively. From pig feeds, 61% of the A. niger var. awamori were producers of this toxin with mean levels of 16 ng ml−1. In poultry feeds, the lowest percentage of OA producer strains was detected. The results for the occurrence of OA in feeds from different sampling months depended on storage and humidity-temperature conditions. Therefore, a good storage practice becomes very important to prevent OA production

Determination of inosine 5′‐monophosphate and guanosine 5′‐monophosphate in pig feed by capillary zone electrophoresis

AbstractA capillary zone clectrophoresis method was developed for the determination of IMP and GIMP, commonly used as flavor enhancers in poultry feed, in a real sample of complex composition. A baseline separation of inosine 5′‐monophosphate and guanosine 5′‐monophosphate was achieved within 10 min and the other components in the sample did not interfere with the separation. Quantitative results obtained from pig feed samples are presented. The separation conditions and experimental reproducibility are also discussed.

Indirect enzyme-linked immunosorbent assay for the mycotoxin zearalenone.

A competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of zearalenone, an estrogenic mycotoxin. Zearalenone was converted to zearalenone-6'-carboxymethyloxime and conjugated to bovine serum albumin and poly-L-lysine for use as immunogen and solid-phase marker, respectively. Immunization of rabbits with the bovine serum albumin conjugate resulted in zearalenone antibody titers of 20,480 in 11 weeks. A competitive indirect ELISA was conducted by simultaneously incubating zearalenone with zearalenone antiserum over zearalenone-6'-carboxymethyloxime poly-L-lysine solid phase and then determining the bound rabbit immunoglobulin with goat anti-rabbit peroxidase conjugate. Response range for zearalenone in the resulting competition curve was between 1 and 50 ng/ml. Reactivities of this antiserum for alpha-zearalenol, beta-zearalenol, alpha-zearalanol, and beta-zearalanol were, respectively, 50, 12, 6, and 3% of that found for zearalenone. By using the competitive indirect ELISA, zearalenone was detectable in methanol-water extracts of corn, wheat, and pig feed samples.