A simple HPLC method for the determination of the mycotoxins ochratoxin A and B in blood serum of swine.

This paper presents a simple method for the determination of ochratoxins A (OTA) and B (OTB) in pig blood serum. The method includes serum acidification (pH < 1.6) and precipitation of protein with 15% trichloroacetic acid, liquid partitioning with dichloromethane and fluorescence detection. The estimated detection limits were 0.1 ng OTA/ml and 0.2 ng OTB/ml. The mean recoveries from artificially contaminated samples (n = 6 replicates/mycotoxin) spiked at 0.3, 1 and 3ng OTA and OTB/ml, respectively, were 86.8% (s.d. = 8.4) for OTA and 90.0% (s.d. = 9.8) for OTB. Forty-nine Romanian pig blood serum samples (94% of 52 analysed) were found to be naturally contaminated with OTA in the range 0.1-13.4 ng/ml. No sample was found positive for OTB. The method is technically simple, specific, cost effective, suitable for large sample throughput and requires small amount of sample and reagents. It fulfils the criteria for a routine method and could be a suitable toolfor surveying OTA in pig herds and in slaughtered pigs.

Survey of Romanian slaughtered pigs for the occurrence of mycotoxins ochratoxins A and B, and zearalenone.

Blood serum, kidney, liver and muscle sample per animal were collected from slaughtered pigs (n = 52). The samples were analysed for ochratoxin A (OTA) and B (OTB) by HPLC methods. Zearalenone (ZEA) in serum was analysed by enzyme immunoassay. A total of 98% serum samples were OTA positive in the range of 0.05-13.4 ng/ml and 85% contained under 5 ng OTA/ml. The incidences of OTA in kidney and liver were very similar (79%, 75%) with mean levels of 0.54 ng/g and 0.16 ng/g, respectively. The lowest incidence (17%) and the lowest mean level contamination (0.15 ng/g) were in muscle samples. The mean distribution in tissues followed the pattern serum > kidney > liver > muscle (100%; 0.26%; 8.5%; 2.57%). No kidney, liver or muscle sample was found OTA positive above the maximum admitted limit in Romania (5 ng/g). No sample was found to be positive for OTB. A very similar OTA contamination (mean = 4.19 ng/ml, coefficient of variation = 34.4%) was observed in the serum samples (n = 10) collected from the same farm. A possible difference in regional distribution of OTA in Romania is suggested. Zearalenone was detected only in 17.3% of the serum samples with a maximum concentration of 0.96 ng/ml. This study shows the presence of OTA and ZEA in Romanian slaughtered pigs at levels comparable to those reported in other countries.

Effects of feeding aFusarium poae extract and a natural zeolite to broiler chickens.

A feeding trial was conducted in order to determine the effects of aFusarium poae extract on the health and performances of broiler chickens and the possible protective effect of a natural zeolite. TheF. poae extract contained nivalenol, T-2 toxin and diacetoxyscirpenol and demonstrated high toxicity when administeredi.p. to rats. One-day-old broiler chickens were fedad libitum over a period of 28 days with the following diets: group I - control; group II - 0.5% zeolite; group III -F. poae extract; group IV-0.5% zeolite andF. poae extract. Broilers were sacrificed at 28 days for the measurement of relative organs weights, leukocyte counts and serum biochemical values. No mortality was recorded over the experiment. Body weight gains, feed intake, feed utilisation and water consumption were depressed by theF. poae extract (p<0.05). A decrease of these parameters were also observed in group IV which received the diet with zeolite and theF. poae extract. No significant differences were seen in group II when compared to control. In groups III and IV the relative weights of liver, kidney, hearth and gizzard were significantly increased (p<0.05), while in group II only the relative liver weight was increased.F. poae extract, administered singly or in combination with zeolite, significantly decreased leukocytes count, serum total protein and serum albumin. Zeolite andF. poae extract, singly or combined, increased serum creatinine and uric acid concentrations (p<0.05).These findings indicate that sublethal doses of F. poae extract can affect adversely the performances and the health in broiler chickens. By adding zeolite these impairments could not diminished and for some parameters the zeolite additive increased the adverse effects of the F. poae extract.