Evaluacion del efecto de anabolicos de crecimiento (Zeranol over, Overmax L.A. Premium y Zeranol implante) en novillos de engorde en un periodo de cien dias en la finca El Rastro, El Coral, Chontales

Para el estudio se utilizaron 68 novillos producto del cruce de la raza Pardo Suizo x Brahman con un peso inicial de 230 ± 33 kg, con el objetivo de evaluar el efecto de tres tratamientos anabólicos sobre la ganancia de peso total en pastoreo libre durante un periodo de 100 días, siendo los tratamientos los siguientes: T 1 (Zeranol implante: Zeranol Over®), T2 (Zeranol Over® formulación tixotrópico), T3. (Overmax® LA Premium: Zeranol + ivermectina 3.15%), comparado con el T4 Testigo o Control (Sin implante). A los animales pesados y organizados en tres grupos con características semejantes, se les aplicó los tratamientos y cada grupo estuvo conformado por 17 novillos; de igual manera, fueron desparasitados con Vermectina La Premium 3. 15 %, a razón de 1ml por cada 50 kg de peso vivo (kg p.v), antes de aplicar los agentes anabólicos. Se determinó el incremento en peso a partir del peso final e inicial, y se calculó la Ganancia Media Diaria (GMD) en cada uno de los tratamientos. Para el diseño de dos vías se utilizó el test de Fisher (Análisis de Varianza) y el Test de Friedman, y categorización estadística mediante la Mínima Diferencia Significativa (DMS, P=O.OS). El análisis estadístico determinó efecto altamente significativo (P=O.OOl) en la GMD, en donde la GMD fue de 739, 624, 590 y de 536 gramos para los tratamientos T3 (Overeas® LA Premium: Zeranol + ívermectina), TI (Zeranol implante: Zeranol Over®), T2 (Zeranol Over'l formulación tixotrópica) y el Control, respectivamente. El implante Overmax® LA Premium (Zeranol+ivermectina3.15%) superó en más de 27 % al tratamiento sin implante, siendo este el tratamiento de mayor rentabilidad, con 83.43 dólares en ganancia de peso por animal, y una relación Beneficio/Costo de 1.27 por cada unidad monetaria invertida.

Comparación de zeranol tixotrópico1% vs zeranol 1% más ivermectina 3.15%, y sus efectos sobre la ganancia media diaria y carga parasitaria en terneros de la raza Reyna, finca Santa Rosa, Managua

El propósito del presente trabajo fue comparar el efecto de Zeranol tixotrópico 1% y Zeranol + ivermectina 3.15% sobre la ganancia de peso y carga parasitaria en terneros de la raza Reyna. Se utilizaron 21 terneros con peso de 86.3 ± 0.8 kg y edad de 12.24 ± 2.0 m, agrupados en un diseño completamente al azar (DCA), distribuidos en tres tratamientos T1: Ivermectina, T2: Zeranol tixotrópico 1%, T3: Zeranol + ivermectina 3.15% con 7 repeticiones por tratamiento. Las variables productivas estudiadas fueron: Ganancia media diaria (GMD), Peso final (PF), Ganancia total de peso (GTP); y medidas zoométricas: Perímetro toráxico (PT), Perímetro abdominal (PA), altura a la cruz (AC) y longitud corporal (LC). Los datos fueron analizados por PROC: GLM del paquete estadístico SAS® Ver. 9.1.2. y la comparación de medias por la prueba de Tuckey. Los resultados demuestran que el T1 obtuvo una GMD de 277.95 g, superando a T2 y T3 (222.83g y 265.53g, respectivamente); el T1 obtuvo mayor peso final que el T2 (112.7 kg vs 104.80 kg), pero fue ligeramente superior al T3 (112.7 kg vs 112.00 kg); para GTP el T1 (25.57 kg) y T3 (24.43 kg) fueron superiores al T2 (20.50 kg). Respecto a las medidas zoométricas PT, PA, AC, LC, los mayores valores fueron para el T1 seguido del T3 y T2. Para control de cargas parasitarias, el T3 fue efectivo para Trichuris y Strongylus, el T1 ejerció mejor control para Trichuris, en cambio el T2 fue el de menor control. El análisis financiero favorece al tratamiento T1 por ser de menor costo, sin embargo el T3 manifestó mejor comportamiento en la ganancia de peso a lo largo del estudio, por lo cual este puede ser utilizado a pesar de tener un mayor costo en relación al T1.

Evaluación del efecto del Zeranol implante Ralgro (pellet) vs. Zeranol tixotrópico (en solución) como promotores de crecimiento en novillos de finalización del Centro Integral de Investigación, Innovación, Producción, Extensión y Enseñanza Agropecuaria, Las Lomas durante el período de Abril a Julio 2014

El presente estudio se realizó con el objetivo de generar información útil sobre las diferencias obtenidas en las ganancias de peso como en utilidades generadas por los tratamientos. Utilizando anabólicos con el mismo principio activo (zeranol), pero con distintas presentación comercial como lo es el zeranol Implante Ralgro: pellets y el zeranol Tixotropico: en solución. El estudio fue experimental, utilizando 2 tratamientos y un grupo control. De manera que el tratamiento 1: Zeranol Implante Ralgro y el tratamiento 2: Zeranol Tixotropi co, al seleccionar los novillos se realizo de un grupo de 250 animales, de los cuales se seleccionaron 27 novillos con pesos similares de aproximadamente 350 kg, características raciales de cruces como brahmán con pardo y brahmán con holstien. El diseño que se utilizó fue completamente aleatorio, para separar los novillos en los grupos experimentales se dividieron 9 novillos por cada grupo, estos en 3 unidades experimentales de 3 animales cada una, luego de ser seleccionados y divididos en sus grupos se les aplico los debido tratamientos a evaluar, a partir de ese momento se inició a realizar pesajes cada 15 días hasta cumplir los 90 días que es el periodo de retiro de los tratamientos; obteniendo así una GMD con el tratamiento 1: zeranol implante 0.76 kg /día/animal, con el tratamiento 2: zeranol Tixotropico 0.71 kg/día/animal y con el grupo control: 0.67 kg/día/a nimal. En la relación beneficio/ costo, se obtuvo con el tratamiento1: zeranol implante 1.27 tratamiento 2: zeranol tixotrópico 1.26 y grupo control: 1.25 teniendo mejores utilidades el tratamiento1. Zeranol implante que por cada córdoba invertido obtuvo 0.27 córdobas de ganancia (utilidades).

Development and validation of dry reagent time-resolved fluoroimmuno assays for zeranol and alpha-zearalenol to assist in distinguishing zeranol abuse from Fusarium spp. toxin contamination in bovine urine

Zeranol, an oestrogenic growth promoter in food animals, is banned within the European Union (EU). However, commercially available immunoassay kits for zeranol cross-react with toxins formed by naturally occurring Fusarium spp. fungi, leading to false-positive screening results. This paper describes the validation of a specificity enhanced, rapid dry reagent time-resolved fluoroimmunoassay (TR-FIA) for zeranol (recovery 99%, limit of detection 1.3 ng ml(-1)) demonstrating that up to 150 ng ml(-1) of Fusarium spp. toxins in urine do not lead to false-positive results. This assay will assist EU Member States to implement Council Directive 961 23\EC, which requires states to monitor for potential abuses of zeranol. A similar TR-FIA for the Fusarium spp. toxin a-zearalenol, using the same sample extract, is also described (recovery 68%, limit of detection 5.6 ng ml(-1)). Only the addition of diluted sample extract is required to perform these dry-reagent TR-FIAs, the results being available within 1 h of extract application. The EU-funded project 'Natural Zeranol' (FAIR5-CT97-3443) will use these fluoroimmunoassays to screen bovine urine in four Member States to gather data on the seasonality of Fusarium spp. toxin contamination of urine and the incidence of zeranol screening test positives.

A specificity-enhanced time-resolved fluoroimmunoassay for zeranol employing the dry reagent all-in-one-well principle

A simple dry chemistry time-resolved fluorescence immunoassay (TR-FIA) method was developed for the measurement of zeranol in bovine urine samples. The samples were purified by immunoaffinity chromatography and a specificity-enhanced zeranol antibody was employed in the immunoassay. This resulted in a highly selective method, which had only negligible reactivity with Fusarium spp, toxins. The all-in-one-well dry chemistry concept made the assay very simple to use because all the assay-specific reagents were already present in the reaction wells in dry form. Only the addition of diluted sample extract was required to perform the competitive one-step TR-FIA and the results were available in less than 1 h. The analytical limit of detection (mean + 3s) for the immunoassay was 0.16 ng ml(-1) (n=12) and the functional limit of detection for the whole method, estimated by the analysis of zeranol-free samples, was 1.3 ng ml(-1) (n=20). The recovery of zeranol at the level of 2 ng ml(-1) was 99% (n=18) and the within-assay variation ranged between 4.5 and 9.0%.

POSSIBLE NATURALLY-OCCURRING ZERANOL IN BOVINE BILE IN NORTHERN-IRELAND

Zeranol and two Fusarium toxins, alpha-zearalenol and beta-zearalenol, were confirmed by gaschromatography/mass spectrometry (GC/MS) in bovine bile samples referred to this laboratory for analysis. No evidence of zeranol abuse was found on-farm. Given the recent suggestion that zeranol might arise from the metabolism of these Fusarium toxins, and the finding of zeranol in bovine and ovine urine across the EU, it. was concluded that the residues had arisen as a result of natural metabolism.

Interlaboratory ring test of time-resolved fluoroimmunoassays for zeranol and alpha-zearalenol and comparison with zeranol test kits

Many zeranol immunoassay test kits cross-react with toxins formed by naturally occurring Fusarium spp. fungi, leading to false-positive screening results. This paper describes the evaluation and application of recently published, dry reagent time-resolved fluoroimmunoassays (TR-FIA) for zeranol and the toxin alpha-zearalenol. A ring test of bovine urine fortified with zeranol and/or alpha-zearalenol in four European Union National Reference Laboratories demonstrated that the TR-FIA tests were accurate and robust. The alpha-zearalenol TR-FIA satisfactorily quantified alpha-zearalenol in urine fortified at 10-30 ng ml(-1) . The specificity-enhanced zeranol TR-FIA accurately quantified zeranol in the range 2-5 ng ml(-1) and gave no false-positive results in blank urine, even in the presence of 30 ng ml(-1) alpha-zearalenol. Zeranol TR-FIA specificity was demonstrated further by analysing incurred zeranol-free urine samples containing natural Fusarium spp. toxins. The TR-FIA yielded no false-positive results in the presence of up to 22 ng ml(-1) toxins. The performance of four commercially available zeranol immunoassay test kits was more variable. Three kits produced many false-positive results. One kit produced only one potential false-positive using a protocol that was longer than that of the TR-FIA. These TR-FIAs will be valuable tools to develop inspection criteria to distinguish illegal zeranol abuse from contamination arising from in vivo metabolism of Fusarium spp. toxins.

The appraisal of an automated multi-immunoaffinity chromatography system to detect anabolic agents in bile and urine

Screening for residues of anabolic steroids frequently requires extraction from tissues and fluids before analysis. Chemical procedures for these extractions can be complicated, expensive to perform and not ideal for the simultaneous extraction of analytes with different solubilities. Extraction by multi-immunoaffinity chromatography (MIAC) may be used as an alternative. Samples are passed through a column containing a range of antibodies immobilized on an inert support. The desired analytes are bound to their respective antibodies, washed and then eluted by a suitable solvent. The purified extracts can then be incorporated into the analytical tests, The analytes that can be extracted presently are alpha-nortestosterone, zeranol, trenbolone, diethylstilboestrol, boldenone and dexamethasone. Manually, the MIAC procedure is limited to about six columns per operator but bq automating the process using a robotic sample processor (RSP), 48 columns can be run simultaneously during the day or night. The RSP has also been adapted to transfer extracts and reagents on to ELISA plates. The automated system has proved to be a robust and reliable means of screening large numbers of samples for anabolic agents with minimal manual input

Specificity enhancement of polyclonal antisera by the induction of tolerance to unwanted cross-reacting determinants

Steroids form a structurally closely related group. As a result, antibodies produced for use in immunoassays regularly show unwanted cross-reactivities, These may be reduced by altering hapten-protein coupling procedures, thereby reducing the exposure of the determinants giving rise to the undesirable cross-reaction. However, these procedures carl prove to be complex, expensive and nor totally predictable in outcome. Exploitation of the clonal selection theory is an attractive alternative approach. The host is primed with the interfering cross-reactant coupled to a non-immunogenic amino acid copolymer to inactivate the B-lymphocyte clones specific for this steroid, producing a specific immunotolerance. Then, 3 days Inter, the host is immunized with the steroid against which nn antibody is required. The clones producing antibody to this immunogen are unaffected and the cross-reactivity is significantly reduced or deleted The technique has been applied to the reduction of endogenous sex steroid cross-reactivity from antibodies prepared against synthetic and semi-synthetic androgens (17 alpha-methyltestosterone, 19-nor-beta-testosterone) and the progestogen medroxyprogesterone. Antibodies prepared against the synthetic oestrogen zeranol using this technique have significantly reduced its undesirable cross-reactivity with the fungal metabolite 7 alpha-zearalenol. Highly specific antisera have been generated in all cases, the only adverse effect being a reduction in the titres achieved in comparison with rabbits receiving the conventional immunizing regime.