Development of Immunoassay for Detection of Meat and Bone Meal in Animal Feed

2005 ◽  
Vol 68 (9) ◽  
pp. 1860-1865 ◽  
Author(s):  
SHIN-HEE KIM ◽  
TUNG-SHI HUANG ◽  
THOMAS A. SEYMOUR ◽  
CHENG-I WEI ◽  
STEPHEN C. KEMPF ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Monoclonal Antibodies ◽  
Animal Feed ◽  
Superior Performance ◽  
Hybridoma Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
Efficient Detection ◽  
Heat Treated

An immunoassay system was developed for efficient detection of prohibited meat and bone meal (MBM) in animal feed. Monoclonal antibodies (MAbs) were raised against bovine smooth muscle autoclaved at 130°C for 20 min. Among the 1,500 supernatants of hybridoma cells screened, MAbs 3E1, 1G3, and 3E10 were selected and characterized in this study. The first set of MAbs produced, 3E1 and 1G3, had stronger reactivity against MBM than against smooth muscle that was heat treated at 90°C for 10 min. However, reactivity gradually increased against smooth muscle that was autoclaved at 130°C for up to 1 h. The enzyme-linked immunosorbent assay for detection of MBM in animal feed was optimized with the MAb 3E10 because of its superior performance. MAb 3E10 diluted to 100-fold was used to differentiate bovine MBM from that of other species in ingredients used for commercial animal feeds and could detect down to 0.05% MBM mixed in animal feed.

scholarly journals Development of Opsonic Mouse Monoclonal Antibodies against Multidrug-Resistant Enterococci

2019 ◽  
Vol 87 (9) ◽  
Author(s):  
Ermioni Kalfopoulou ◽  
Diana Laverde ◽  
Karmela Miklic ◽  
Felipe Romero-Saavedra ◽  
Suzana Malic ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Capsular Polysaccharide ◽  
Selection Process ◽  
Multidrug Resistant ◽  
Expression Vectors ◽  
Hybridoma Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Strains ◽  
Variable Regions ◽  
Content Type

ABSTRACTMultidrug-resistant enterococci are major causes of hospital-acquired infections. Immunotherapy with monoclonal antibodies (MAbs) targeting bacterial antigens would be a valuable treatment option in this setting. Here, we describe the development of two MAbs through hybridoma technology that target antigens from the most clinically relevant enterococcal species. Diheteroglycan (DHG), a well-characterized capsular polysaccharide ofEnterococcus faecalis, and the secreted antigen A (SagA), an immunogenic protein fromEnterococcus faecium, are both immunogens that have been proven to raise opsonic and cross-reactive antibodies against enterococcal strains. For this purpose, a conjugated form of the native DHG with SagA was used to raise the antibodies in mice, while enzyme-linked immunosorbent assay and opsonophagocytic assay were combined in the selection process of hybridoma cells producing immunoreactive and opsonic antibodies targeting the selected antigens. From this process, two highly specific IgG1(κ) MAbs were obtained, one against the polysaccharide (DHG.01) and one against the protein (SagA.01). Both MAbs exhibited good opsonic killing against the target bacterial strains: DHG.01 showed 90% killing againstE. faecalistype 2, and SagA.01 showed 40% killing againstE. faecium11231/6. In addition, both MAbs showed cross-reactivity toward otherE. faecalisandE. faeciumstrains. The sequences from the variable regions of the heavy and light chains were reconstructed in expression vectors, and the activity of the MAbs upon expression in eukaryotic cells was confirmed with the same immunological assays. In summary, we identified two opsonic MAbs against enterococci which could be used for therapeutic or prophylactic approaches against enterococcal infections.

Validation of a PCR-Based Method for the Detection of Various Rendered Materials in Feedstuffs Using a Forensic DNA Extraction Kit

2006 ◽  
Vol 69 (1) ◽  
pp. 205-210 ◽  
Author(s):  
MICHAEL J. MYERS ◽  
HAILE F. YANCY ◽  
MICHAEL ARANETA ◽  
JENNIFER ARMOUR ◽  
JANICE DERR ◽  
...  
Keyword(s):  
False Positive ◽  
Animal Feed ◽  
Limit Of Detection ◽  
False Positive Rate ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
Average Accuracy ◽  
Positive Rate ◽  
Feed Samples ◽  
Pcr Primer

A method trial was initiated to validate the use of a commercial DNA forensic kit to extract DNA from animal feed as part of a PCR-based method. Four different PCR primer pairs (one bovine pair, one porcine pair, one ovine primer pair, and one multispecies pair) were also evaluated. Each laboratory was required to analyze a total of 120 dairy feed samples either not fortified (control, true negative) or fortified with bovine meat and bone meal, porcine meat and bone meal (PMBM), or lamb meal. Feeds were fortified with the animal meals at a concentration of 0.1% (wt/wt). Ten laboratories participated in this trial, and each laboratory was required to evaluate two different primer pairs, i.e., each PCR primer pair was evaluated by five different laboratories. The method was considered to be validated for a given animal source when three or more laboratories achieved at least 97% accuracy (29 correct of 30 samples for 96.7% accuracy, rounded up to 97%) in detecting the fortified samples for that source. Using this criterion, the method was validated for the bovine primer because three laboratories met the criterion, with an average accuracy of 98.9%. The average false-positive rate was 3.0% in these laboratories. A fourth laboratory was 80% accurate in identifying the samples fortified with bovine meat and bone meal. A fifth laboratory was not able to consistently extract the DNA from the feed samples and did not achieve the criterion for accuracy for either the bovine or multispecies PCR primers. For the porcine primers, the method was validated, with four laboratories meeting the criterion for accuracy with an average accuracy of 99.2%. The fifth laboratory had a 93.3% accuracy outcome for the porcine primer. Collectively, these five laboratories had a 1.3% false-positive rate for the porcine primer. No laboratory was able to meet the criterion for accuracy with the ovine primers, most likely because of problems with the synthesis of the primer pair; none of the positive control DNA samples could be detected with the ovine primers. The multispecies primer pair was validated in three laboratories for use with bovine meat and bone meal and lamb meal but not with PMBM. The three laboratories had an average accuracy of 98.9% for bovine meat and bone meal, 97.8% for lamb meal, and 63.3% for PMBM. When examined on an individual laboratory basis, one of these four laboratories could not identify a single feed sample containing PMBM by using the multispecies primer, whereas the other laboratory identified only one PMBM-fortified sample, suggesting that the limit of detection for PMBM with this primer pair is around 0.1% (wt/wt). The results of this study demonstrated that the DNA forensic kit can be used to extract DNA from animal feed, which can then be used for PCR analysis to detect animal-derived protein present in the feed sample.

scholarly journals Meat and bone meal still used in animal feed

Nature ◽  
2001 ◽  
Vol 414 (6860) ◽  
pp. 147-147
Author(s):  
Stephen Rossides
Keyword(s):  
Animal Feed ◽  
Meat And Bone Meal ◽  
Bone Meal

Detection of Rendered Meat and Bone Meals by PCR Is Dependent on Animal Species of Origin and DNA Extraction Method

2010 ◽  
Vol 73 (6) ◽  
pp. 1090-1096 ◽  
Author(s):  
MICHAEL J. MYERS ◽  
DOROTHY E. FARRELL ◽  
CHRISTINE M. DEAVER ◽  
JACQULINE MASON ◽  
HEIDI L. SWAIM ◽  
...  
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Animal Species ◽  
Animal Feed ◽  
Functional Assay ◽  
Threshold Values ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
Pcr Method

The capability of eight commercially available DNA extraction kits to extract bovine DNA originating in meat and bone meal from fortified feed was evaluated. Four different batches of bovine meat and bone meal (BMBM) were used for DNA extraction with the eight commercial DNA extraction kits. Within each kit, there were minimal differences in the batch-to-batch amounts of extracted DNA. There were differences between the kits in the amounts of DNA that could be extracted from the same amount of starting BMBM. These differences did not translate into differences in the amount of amplifiable DNA from BMBM-fortified dairy feed. Using a validated real-time PCR method, the kit yielding the highest amount extractable DNA was completely unable to yield a positive PCR result; one other kit was also unable to produce a positive PCR result from DNA extracted from BMBM-fortified feed. There was a complete lack of a correlation between the amount of bovine DNA isolated from BMBM by a given extraction kit compared with the relative amounts of DNA isolated from fortified animal feed as evidenced by the cycle threshold values generated using the real-time PCR method. These results demonstrate that extraction of DNA from processed animal protein is different for pure ingredients and fortified animal feeds. These results indicate that a method specifically developed using just animal-derived meat and bone meal may not yield a functional assay when used to detect animal tissues in complete animal feed.

Production of species-specific monoclonal antibodies that react with surface components on zoospores and cysts of Phytophthora nicotianae

1998 ◽  
Vol 44 (12) ◽  
pp. 1161-1170 ◽  
Author(s):  
A V Robold ◽  
A R Hardham
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Phytophthora Cinnamomi ◽  
Phytophthora Nicotianae ◽  
The Other ◽  
Hybridoma Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Hybridoma Cell Lines ◽  
Species Specific

Monoclonal antibodies were generated against components on the surface of zoospores and cysts of the Oomycete, Phytophthora nicotianae, with the aim of obtaining antibodies diagnostic for this species of plant pathogen. A dipstick version of an enzyme-linked immunosorbent assay was used to screen hybridoma cell lines produced by following a coimmunization protocol in which a mouse was immunized with Phytophthora nicotianae cysts mixed with murine antisera raised against cysts of Phytophthora cinnamomi and Phytophthora cryptogea. Of the nine hybridoma cells lines which remained positive, five produced antibodies that reacted with species-specific epitopes on the surface of the spores. Immunofluorescence, immunogold, and immunoblot labelling showed that three of the five species-specific antibodies reacted with a polypeptide of relative molecular mass greater than 205 kDa which was distributed over the entire zoospore surface, including that of the two flagella. These antibodies also labelled the surface of cysts to varying degrees. The other two species-specific antibodies bound to the shaft of tubular mastigonemes that form two rows on the anterior flagellum. In immunoblots, one of these antibodies recognised a 40-kDa glycoprotein. Antibodies produced by the other four hybridoma cell lines reacted with all Phytophthora and Pythium species tested. The results (i) showed that the coimmunization technique effectively produced antibodies directed towards components specific for Phytophthora nicotianae in the presence of antigens common to many Phytophthora species, and (ii) revealed for the first time the biochemical nature of molecular constituents of flagellar mastigonemes in the Oomycetes.Key words: cell surface, flagella, immunodiagnostics, mastigonemes, monoclonal antibodies.

Production of Monoclonal Antibody for the Detection of Meat and Bone Meal in Animal Feed

2004 ◽  
Vol 52 (25) ◽  
pp. 7580-7585 ◽  
Author(s):  
Shin-Hee Kim ◽  
Tung-Shi Huang ◽  
Thomas A. Seymour ◽  
Cheng-i Wei ◽  
Stephen C. Kempf ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Animal Feed ◽  
Meat And Bone Meal ◽  
Bone Meal

Development of the ELISAs for detection of hormone-disrupting chemicals

2000 ◽  
Vol 42 (7-8) ◽  
pp. 81-88 ◽  
Author(s):  
Y. Goda ◽  
A. Kobayashi ◽  
K. Fukuda ◽  
S. Fujimoto ◽  
M. Ike ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Phthalate Esters ◽  
Hybridoma Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spleen Cells ◽  
Myeloma Cells ◽  
Structural Resemblance ◽  
Reaction Test ◽  
Mouse Myeloma

Six kinds of enzyme-linked immunosorbent assay (ELISA) systems were developed for the quantitative analysis of hormone-disrupting chemicals (HDCs), such as estrogen (ES: the total amount of estrone (E1), 17 β-estra (E2) and estriol (E3)), E2, bisphenol A (BPA), alkylphenol (AP), phthalate esters (PE) and chlorophenols (CP). To generate specific monoclonal antibodies against BPA, AP, PE, CP, hybridoma cells were produced by the fusion of mouse myeloma cells and spleen cells from mice immunized with carboxylated derivatives, while anti E2 monoclonal antibody was selected from those available on the market, and anti ES monoclonal antibody was purchased from Teikoku Hormone Mfg Co. Ltd. The detection limits of ES, E2, BPA, AP, PE and CP ELISAs were 0.1, 0.1, 5, 10, 200, 10 μg/L, when E2, E2, BPA, Nonylphenol (NP), Dibutylphthalate (DBP), 2,4-CP were used as standard, respectively, and the specificity of each ELISA was confirmed with the cross-reaction test using several compounds which have structural resemblance to the compounds of interest.

The concentration of strontium and other minerals in animal feed ingredients

2013 ◽  
Vol 2 ◽  
Author(s):  
L.C. Browning ◽  
A.J. Cowieson
Keyword(s):  
Inductively Coupled Plasma ◽  
Animal Feed ◽  
Optical Emission ◽  
Grain Legume ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
High Concentration ◽  
Mineral Concentration ◽  
Feed Ingredients ◽  
Cereal Grain

SummaryVariance in macro- and micro-mineral concentration in feed ingredients for farmed livestock contributes to sub-optimal performance and may compromise health and welfare. Although routine quality assurance and quality control procedures in feed mills or integrated poultry or swine businesses may track variance in the concentration of minerals of immediate nutritional importance, such as phosphorus (P), calcium (Ca) and sodium (Na), micro-minerals such as strontium (Sr) attract less attention. In order to create a framework for further study, the mineral concentration in more than 130 animal feed ingredients commonly used in Australia were analysed by inductively coupled plasma optical emission spectroscopy (ICP-OES). Due to a dearth of information, the principal focus of the survey was Sr, but the concentration of Ca, P, magnesium (Mg), manganese (Mn), potassium (K), iron (Fe), copper (Cu), zinc (Zn), sulphur (S) and Na were analysed concurrently. Generally the minerals present at the highest concentrations in the various feed ingredients examined were Ca, P and Mg. As anticipated, the ingredients with the highest concentrations of Ca and P were inorganic phosphates, limestone and meat and bone meal. The average Ca concentration in limestone was 393 g/kg but a range of 376–415 g/kg was observed which may be nutritionally important. Furthermore, the Mg concentration in limestone ranged from 7–535 mg/kg suggesting some contamination by dolomite lime sources. A total of 24 meat and bone meal samples were included in the analysis and mean Ca and P concentrations were 109 and 54 g/kg respectively. However, the range of Ca and P in meat and bone meal was considerable with Ca concentrations from 51–148 g/kg and P concentrations from 26–66 g/kg. A total of 81 cereal, grain legume and cereal by-product samples were included as part of the survey and these vegetable feed ingredients contained relatively low concentrations of most minerals with Ca, P, Mg and K dominating. The K concentration of soybean meal was found to be around 23 g/kg and ranged from approximately 22–27 g/kg. In comparison, the Sr concentration in the feed ingredients was low relative to other minerals, with limestone having the highest level of strontium at 329 mg/kg. Overall those feed ingredients from a mineral origin had the highest level of Sr. In addition, meat and bone meal had a relatively high concentration of Sr (around 159 mg/kg).

Comparison between fish meal and two Queensland meat and bone meals in bacon pig production

1965 ◽  
Vol 5 (19) ◽  
pp. 404 ◽  
Author(s):  
ACE Todd ◽  
LJ Daniels
Keyword(s):  
Crude Protein ◽  
Fish Meal ◽  
Grain Sorghum ◽  
Superior Performance ◽  
Food Conversion ◽  
Meat And Bone Meal ◽  
Bone Meal ◽  
Significant Difference ◽  
Lower Protein ◽  
Daily Gain

Fish meal, containing 64 per cent crude protein, was fed at a level of 10 per cent, as a protein supplement, in rations based on grain sorghum. Food conversion and daily gain of pigs fed this diet, from shortly after weaning to bacon weight, was superior to that obtained on diets containing either 14 per cent of a 65 per cent or 16 per cent of a 49 per cent crude protein meat and bone meal. Superior performance was also obtained when this diet was compared with two mixtures of the lower protein meat and bone and fish meal. There was no significant difference in performance when the fish meal diet was compared with a mixture of the higher protein meat and bone and fish meal. In spite of its high cost the imported fish meal was a very economical supplement.

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