Monitoring and identification of residues of anabolic preparations in slaughtered cattle by HPLC with diode array detection

Summary

The new combination of isocratic high performance liquid chromatography (HPLC) with on line UV spectrum detection via a diode array configuration has been applied to the detection and identification of anabolics present in application sites of cattle.

Combination of the characteristic retention time in the HPLC chromatogram and a comparison of the full spectrum between 190–400 nm of the anabolic components with that of a standard resulted in a very reliable identification. By means of this method 117 samples of application sites were investigated for the presence of anabolic residues. Of the xenobiotic anabolics, 19‐nortestosterone (NT) was found most frequently (in 96 cases), whereas diethylstilbestrol (DES) was found in only 11 cases.

In all samples the identification of NT and DES was confirmed by high resolution gas chromatography‐mass spectrometry (GCMS).     

Experimental studies on the impact of an increased dose of anionic salts on the metabolism of dairy cows

Summary

This study was initiated to investigate the influence of a daily dose of anionic salts (AS) above the valid upper limit at present on metabolism of dairy cows. Eleven non‐pregnant and non‐lactating Holstein‐Friesian crossbred cows with a permanent rumen cannula were used in a study with a controlled feeding design. The initial daily dose was 2500 meq/day, which resulted in a Dietary Cation Anion Difference (DCAD) of ‐211 meq/kg dry matter. Every seven days, the daily dose was raised by 500 meq. If a cow stopped eating, the application of AS was stopped and these cows were monitored over the next seven days. On day 30 another batch of hay, having the same DCAD but higher concentrations of minerals and energy, was fed. Blood and urine samples were taken to monitor acid‐base balance and calcium concentrations. Acid‐base balance was strongly influenced by AS. Blood pH dropped steadily and reached values around 7.23. Urine pH dropped quickly below 6 and remained at that level regardless of the increased dosage of AS. Net acid base excretion (NABE) fell continuously with the increase of the dosage of AS and reached values below ‐200 mmol/l. Calcium concentrations in the serum were nearly stable, but those in urine increased sharply and remained on an elevated level with increasing doses of AS. A few days before the individual cow's refusal of feed intake, calcium excretion in urine decreased. The majority of cows stopped eating while consuming a diet containing 3500 to 4000 meq AS except two animals who consumed up to 6000 meq/day AS but they received the better hay in the second half of the treatment period. In this time pH in blood increased slowly. NABE remained stable on a low level at ‐100 mmol/l. The results showed that with an increasing amount of AS fed the risk of clinical acidosis increased. The decreasing urine concentrations of calcium indicate a breakdown of the compensation capability of the single cow. Besides the dose of AS fed, the quality of the feed stuff might be another factor concerning the tolerance of cows against AS.     

Impact of blood serum insulin-like growth factor I on platelet-activating factor in bull spermatozoa

The objective of this study was to examine differences in platelet-activating factor [1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine; PAF] in spermatozoa between two lines of Angus beef cattle divergently selected for blood serum insulin-like growth factor I (IGF-I) concentration. Endogenous lipids were extracted from the spermatozoa and endogenous PAF content was determined by radioimmunoassay. The amount of PAF detected in spermatozoa obtained from high IGF-I bulls (n = 8) ranged from 0.145 to 3.571 pM/10(6) cells. The level of PAF extracted from spermatozoa obtained from low IGF-I- bulls (n = 5) ranged from 0.001 to 1.024 pM/10(6) cells. Polynomial regression analysis revealed a significant cubic relationship (R(2) = 0.374; F = 6.292; P < 0.05) between spermatozoa PAF content and blood serum IGF-I concentration. Spermatozoa-derived PAF levels (mean +/- SEM) were significantly higher (P < 0.05) in the high IGF-I group (1.90 +/- 0.39 pM/10(6) cells) than in the low IGF-I group (0.59 +/- 0.20 pM/10(6) cells). High IGF-I bulls have a greater than three-fold higher PAF content in their spermatozoa than low IGF-I bulls. The data demonstrate that not only is PAF present in bull spermatozoa but that levels are significantly higher in individuals with high serum IGF-I concentrations.     

Use of medetomidine and ketamine for immobilization of free-ranging giraffes

OBJECTIVE: To develop a dosage correlated with shoulder height (SH) in centimeters for effective immobilization of free-ranging giraffes, using a combination of medetomidine (MED) and ketamine (KET) and reversal with atipamezole (ATP). DESIGN: Prospective study. ANIMALS: 23 free-ranging giraffes. PROCEDURE: The drug combination (MED and KET) was administered by use of a projectile dart. Quality of induction, quality of immobilization, and time to recovery following injection of ATP were evaluated. Physiologic variables measured during immobilization included PaO2, PaCO2, oxygen saturation, end-tidal CO2, blood pH, indirect arterial blood pressure, heart and respiratory rates, and rectal temperature. RESULTS: Sixteen giraffes became recumbent with a dosage (mean +/- SD) of 143 +/- 29 microg of MED and 2.7 +/- 0.6 mg of KET/cm of SH. Initially, giraffes were atactic and progressed to lateral recumbency. Three giraffes required casting with ropes for data collection, with dosages of 166 +/- 5 microg of MED and 3.2 +/- 0.6 mg of KET/cm of SH. Four giraffes required administration of etorphine (n = 2) or were cast with ropes (2) for capture but remained dangerous to personnel once recumbent, precluding data collection. In giraffes successfully immobilized, physiologic monitoring revealed hypoxia and increased respiratory rates. Values for PaCO2, end-tidal CO2, and heart rate remained within reference ranges. All giraffes were hypertensive and had a slight increase in rectal temperature. Atipamezole was administered at 340 +/- 20 microg/cm of SH, resulting in rapid and smooth recoveries. CONCLUSIONS AND CLINICAL RELEVANCE: Medetomidine and KET was an effective immobilizing combination for free-ranging giraffes; however, at the dosages used, it does not induce adequate analgesia for major manipulative procedures. Quality of induction and immobilization were enhanced if the giraffe was calm. Reversal was rapid and complete following injection of ATP.     

Epidemiology and control of Menangle virus in pigs

OBJECTIVE: To describe the epidemiology and eradication of Menangle virus infection in pigs. DESIGN: Field observations and interventions, structured and unstructured serological surveys, prospective and cross-sectional serological studies and laboratory investigations. PROCEDURE: Serum samples were collected from pigs at a 2600-sow intensive piggery in New South Wales that experienced an outbreak of reproductive disease in 1997. Serum samples were also collected from piggeries that received pigs from or supplied pigs to the affected piggery and from other piggeries in Australia. Serum and tissue samples were collected from pigs at piggeries experiencing reproductive disease in New South Wales. Sera and faeces were collected from grey-headed flying foxes (Pteropus poliocephalus) in the region of the affected piggery. Serum samples were tested for neutralising antibodies against Menangle virus. Virus isolation was attempted from faeces. RESULTS: Following the outbreak of reproductive disease, sera from 96% of adult pigs at the affected piggery, including sows that produced affected litters, contained neutralising antibodies against Menangle virus. Neutralising antibodies were also detected in sera from 88% of finisher pigs at two piggeries receiving weaned pigs from the affected piggery. No evidence of Menangle virus infection was found in other piggeries in Australia. In cross-sectional studies at the affected piggery, colostral antibodies were undetectable in most pigs by 14 to 15 weeks of age. By slaughter age or entry to the breeding herd, 95% of pigs developed high antibody titres (> or = 128) against Menangle virus in the virus neutralisation test. Menangle virus was eradicated from the affected piggery following a program of serological testing and segregation. Neutralising antibodies against Menangle virus were also detected in P poliocephalus from two colonies in the vicinity of the affected piggery. Two piggery workers were infected with Menangle virus. There was no evidence of infection in cattle, sheep, birds, rodents, feral cats and a dog at the affected piggery. CONCLUSIONS: Serological evidence of infection with Menangle virus was detected in pigs at a piggery that had experienced reproductive disease, in pigs at two associated piggeries and in fruit bats in the region of the piggery. Two humans were infected. The mode of transmission between pigs is unknown, but spread by faecal or urinary excretion is postulated. This virus can be eradicated by the segregation of pigs into discrete age groups.     

Genetic correlation estimates between ultrasound measurements on yearling bulls and carcass measurements on finished steers.

Carcass and growth measurements of finished crossbred steers (n = 843) and yearling ultrasound and growth measurements of purebred bulls (n = 5,654) of 11 breeds were analyzed to estimate genetic parameters. Multiple-trait restricted maximum likelihood (REML) was used to estimate heritabilities and genetic correlations between finished steer carcass measurements and yearling bull ultrasound measurements. Separate analyses were conducted to examine the effect of adjustment to three different end points: age, backfat thickness, and weight at measurement. Age-constant heritability estimates from finished steer measurements of hot carcass weight, carcass longissimus muscle area, carcass marbling score, carcass backfat, and average daily feedlot gain were 0.47, 0.45, 0.35, 0.41, and 0.30, respectively. Age-constant heritability estimates from yearling bull measurements of ultrasound longissimus muscle area, ultrasound percentage of intramuscular fat, ultrasound backfat, and average daily postweaning gain were 0.48, 0.23, 0.52, and 0.46, respectively. Similar estimates were found for backfat and weight-constant traits. Age-constant genetic correlation estimates between steer carcass longissimus muscle area and bull ultrasound longissimus muscle area, steer carcass backfat and bull ultrasound backfat, steer carcass marbling and bull ultrasound intramuscular fat, and steer average daily gain and bull average daily gain were 0.66, 0.88, 0.80, and 0.72, respectively. The strong, positive genetic correlation estimates between bull ultrasound measurements and corresponding steer carcass measurements suggest that genetic improvement for steer carcass traits can be achieved by using yearling bull ultrasound measurements as selection criteria.