C.A. Sgoifo Rossi

Neoformation of boldenone and related steroids in faeces of veal calves

Abstract Conflicting findings regarding the boldenone content of bovine faeces suggest it may be synthesized de novo in emitted faeces. We tested this hypothesis by analysing uncontaminated urine, fresh and various forms of dried faeces from 10 calves (not given boldenone) by liquid chromatography/tandem mass spectrometry for 17α- and 17β-boldenone (α and β BOL); 1,4-androstadiene-3,17-dione (ADD); 4-androstene-3,17-dione (AED), testosterone (T) and epitestosterone (ET). Urine contained no α BOL, β BOL or ADD. The analysed substances were variably present in the rectal faeces, and at generally higher levels in faeces scraped from skin or stall floor. In pooled rectal faeces naturally dried for 13 days, α BOL, ADD, AED and ET levels were extremely high (much higher than accounted for by increases due to drying), and β BOL and T were absent. It is concluded that de novo synthesis of α BOL and metabolites occurs naturally in bovine faeces and only uncontaminated urine should be analysed for illegal boldenone.

Evidence for false-positive results for Boldenone testing of veal urine due to faecal cross-contamination during sampling

European Directive 96/22/EC, which controls veterinary residues in animals, does not permit the presence of synthetic growth promoters in products of animal origin or in livestock. Boldenone is categorized in class A3 (growth promoters — steroids) and is thus a banned substance. Testing of veal urine for banned substances is part of the European Union statutory programme for animals going into the food chain. In relation to this monitoring, three studies were conducted to investigate the apparent presence of the banned growth promoter boldenone in veal urine, which was suspected as being caused by interference from faecal contamination of the sample. In the first study, urine samples were collected at different times (time 0 and after 30 min) using (1) a conventional zoonotechnical apron and (2) a technique designed specifically to avoid faecal contamination (‘kettle’). This resulted in samples that were, respectively, positive and negative for the presence of α-boldenone (α-BOL). In a second study, urine samples negative to α-BOL were collected from eight veal calves, but became positive after deliberate faecal contamination. In a third study, data obtained from the Italian RNP (Residual National Program) indicated that 18.1% of 3295 urine samples collected using the zootechnical apron were positive for α-BOL and 2.1% for β-boldenone (β-BOL), whilst of 902 samples collected using the kettle, β-BOL was not detected in any samples and only 0.2% were positive to α-BOL, in concentrations lower than 2 ng ml−1. These results further support the supposition that faecal contamination of the urine during sample collection can lead to false-positive results during boldenone analysis.

Detection of boldenone, its conjugates and androstadienedione, as well as five corticosteroids in bovine bile through a unique immunoaffinity column clean-up and two validated liquid chromatography–tandem mass spectrometry analyses

The presence of β-boldenone II phase metabolites and prednisolone in urine samples, owing to endogenous or natural origin or illicit treatment, is under debate within the European Union. The detection of β-boldenone conjugates, α-boldenone conjugates at concentrations higher than 2 ng mL(-1) and prednisolone above the cut-off level of 5 ng mL(-1) in urine have been, until now, critical in deciding if illegal drug use has occurred. The use of urine sometimes is not entirely satisfactory, especially when the drug is administrated at low doses or when its metabolic conversion is very fast. This subsequently would hamper its detection in urine. The introduction of a new, advantageous matrix where the illicit treatment can be investigated would be highly appreciated. In this study, we have developed and validated a simple and unique immunoaffinity clean-up procedure, which was applied to bovine bile samples, followed by two different analytical liquid chromatography-electrospray-tandem mass spectrometry methods. The first method tests androstadienedione, α- and β-boldenone sulphate, glucuronate and related free forms, while the other method assays prednisolone, prednisone, dexamethasone, cortisone, and cortisol. The methods were validated according to European Commission Decision 2002/657/EC. The evaluated parameters were linearity, specificity, precision (repeatability and intra-laboratory reproducibility), recovery, decision limit and detection capability. The decision limits (CCα) were between 0.38 and 0.45 ng mL(-1) for anabolic steroids, and 0.13 and 0.15 ng mL(-1) as far as corticosteroids were concerned. Intra- and inter-day repeatability was below 15.8 and 19.9% for all analytes, respectively. The methods were applied to the analysis of some bile samples collected from untreated young bulls in order to investigate the presence of the studied steroids in this matrix.

Administration of Essential Oils Cinnamaldehyde, Eugenol, and Capsicum to Beef Cattle: Effects on Health Status and Growth Performance

A 53-day field trial was performed to evaluate the effects of the essential oils of cinnamaldehyde, eugenol, and capsicum on the health status and growth performance of 45 Charolais beef cattle (average arrival weight: 422.0 ± 29.3 kg; control: n = 21; treated: n = 24). Each animal was weighed, and blood samples were collected on days 0, 25, and 53. The average daily gain of the treated group was significantly higher (P < 0.05) than that of the control. The treated group showed a significant increase in antibody for BHV-1 after vaccination (P < 0.05 at day 25 and day 53) and significantly higher serum bactericidal activity (P = 0.01). No differences between groups were observed for serum content of haptoglobin or reactive metabolites of oxygen. The observed improvement in growth performance and health status is due to the capacity of essential oils to optimize rumen fermentations, to increase dry matter intake, and probably to the positive interaction between essential oils and immune system components.

Effect of ageing time in vacuum package on veal longissimus dorsi and biceps femoris physical and sensory traits

Study evaluated the effects of vacuum ageing (2, 4, 6, 8, 10, 12, 16 days) on veal loin (longissimus dorsi; LD) and silverside (biceps femoris; BF) physical and sensory characteristics. Ageing did not affect cooking loss, increased LD pH and L*, a* and b* in both muscles. Shear force (SF) decreased until day 6 in LD and day 10 in BF. Aroma, flavor and taste were not affected, while texture traits were improved. SF was negative correlated with tenderness and juiciness and positive correlated with BF fibrousness and stringy sensation. Ageing improved texture properties without altering other sensory traits.

Determination of malonaldehyde in bovine plasma during the receiving period and effects of phytoderivative diet supplementation

Malonaldehyde (MA) is one of the terminal products of polynsatured fatty acid peroxidation that is considered a useful indicator both in food inspection (Fernandez et al., 1997) and in the chemical-clinical area (Steiner et al., 1996). It is used especially as a marker both of lipid peroxidation and of stress in health field. The receiving period is the most critical period for beef cattle because of the high incidence of diseases, especially those due to stress. Diet administration of natural substances with immunostimulating, tonic and antistress activities, is an efficacious adjuvant in the management of new arrival beef cattle. Such substances can reduce the negative effects of stress which, as is well-known, reduces natural defences against pathogenic agents exposing animals to the risk of considerable microorganism reproduction. The availability of proved stress chemical markers allows reliable evaluation of the efficiency of specific strategies. This study was aimed at evaluating the effects of phytoderivative supplementation on health status in Charolaise beef cattle during the receiving period, using MA as a chemical marker of stress.