Technical note: Development of an enzyme-linked immunosorbent assay for the determination of florfenicol and thiamphenicol in swine feed

One polyclonal antibody against florfenicol and thiamphenicol was produced and a competitive ELISA was developed for the detection of florfenicol and thiamphenicol in swine feed. The ELISA gave a 50% inhibiting concentration of 1.02 ng/mL for florfenicol. For swine feed fortified with 0.05 to 3.0 mg/kg, the interassay recoveries of florfenicol and thiamphenicol ranged from 86.4 to 118.6%, whereas intraassay recoveries of both drug ranged from 90.1 to 126.5% with less than 15% CV. Results obtained from HPLC-tandem mass spectrometry indicated this ELISA procedure could be used as a convenient method for rapid screening of florfenicol and thiamphenicol in swine feed.     

Determination and kinetics of enrofloxacin and ciprofloxacin in Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a liquid chromatography/mass spectrometry method

Determination and kinetics of enrofloxacin and ciprofloxacin in Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a liquid chromatography/mass spectrometry method. J. vet. Pharmacol. Therap. 34, 142-152. The fluoroquinolones enrofloxacin (EF) and ciprofloxacin (CF) residues were investigated in the edible tissues of two important Asian aquacultured species such as Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a sensitive liquid chromatography-electrospray ionization-tandem mass spectrometry method. Fish and prawn were treated with medicated feed with multiple doses of EF, in field conditions. A validation study of the analytical method was realized in terms of linearity, specificity, precision (repeatability and within-laboratory reproducibility), recovery and decision limit (CCα). The time needed before the antibiotic disappears from animal tissues or reach the maximum residue limit (MRL, 100μg/kg) was assessed. The concentration values of EF detected in Tra catfish tissue were between the MRL and 2×MRL concentrations, according to the fish density, 7days following the end of the enrofloxacin treatment (20mg/kg body weight per day, for seven consecutive days). The concentration value of ER in prawn tissue was lower than the MRL and the limit of quantification (LOQ, 14μg/kg) 5 and 7days after the stop of the EF treatment (50mg/kg body weight per day, for five consecutive days), respectively. The mean detected levels of CF was much lower in comparison with that of EF, indicating that only a small part of EF is metabolized into CF (<5%) in both Tra catfish and prawn.     

Effect of long- or short-term feeding of alpha-tocopheryl acetate to Holstein and crossbred beef steers on performance, carcass characteristics, and beef color stability.

Three experiments were conducted to examine the effects of vitamin E supplementation on feedlot cattle. Vitamin E supplementation did not affect feedlot performance or carcass characteristics of cattle fed a high-concentrate diet (P greater than .1). The major finding was the effectiveness of vitamin E in extending the color stability of displayed beef (P less than .01). Color stability during display of longissimus lumborum steaks from cattle supplemented with 300 IU/d for 266 d, 1,140 IU/d for 67 d, or 1,200 IU/d for 38 d was extended by 2.5 to 4.8 d. Gluteus medius steaks had an extended color display life of 1.6 to 3.8 d. The accumulation of lipid oxidation products, but not aerobic microbes, associated with displayed longissimus lumborum was suppressed for muscle from vitamin E-supplemented steers. Taste panelists detected no difference among longissimus lumborum steaks from control and vitamin E-supplemented steers but found (P less than .01) steaks aged for 21 d to be more tender than steaks aged for 7 d. Supplementing cattle with vitamin E should reduce economic losses associated with discolored beef during retail display.     

Associations amongst three feline Porphyromonas species from the gingival margin of cats during periodontal health and disease

Digoxigenin labelled whole chromosomal DNA probes directed against three feline members of the genus Porphyromonas (P. gingivalis VPB 3492, P. circumdentaria NCTC 12469T and P. salivosa VPB 3313) were used to identify and quantify organisms in samples taken from the gingival margins of 40 domestic cats with different grades of periodontal disease. At the right upper canine tooth, the grade of periodontal disease ranged from 0 to 5 and the cfu of facultative/obligate anaerobes ranged from 5.5 x 10(4) to 2.0 x 10(6)). In 38 of the 40 cats, at least one of the three Porphyromonas species was isolated and regression analysis showed that the cfu of total Porphyromonas sp. was a highly significant indicator of the grade of periodontal disease (p < 0.001, R2 0.510). Feline P. gingivalis was isolated from 37 of the 40 cats and regression analysis showed that it was a highly significant predictor of the grade of periodontal disease (p < 0.001, R2 0.561). The cfu of P. salivosa was a significant predictor of the grade of periodontal disease (p < 0.001, R2 0.286) and regression analysis showed that there was a significant positive relationship between cfu of P. circumdentaria and grade of periodontal disease (p = 0.018, R2 0.116). The periodontal grades at the right upper third premolar tooth ranged from 0 to 6. The cfu of facultative/obligate anaerobes isolated ranged from 1.2 x 10(5) to 7.9 x 10(6), and regression analysis showed that cfu was a significant predictor of periodontal grade (p < 0.001, R2 0.378). The cfu of total Porphyromonas species ranged from 1.2 x 10(4) to 1.7 x 10(6) and regression analysis of the cfu against the grade of periodontal disease showed a highly significant association (p < 0.001, R2 0.633). The cfu of P. gingivalis ranged from 0 to 1.1 x 10(6) and regression analysis of the cfu of P. gingivalis against the grade of periodontal disease showed a highly significant association (p < 0.001, R2 0.439). The cfu of P. salivosa was a significant predictor of the grade of periodontal disease (p < 0.001, R2 0.479) and the same association was found between cfu of P. circumdentaria and grade of periodontal disease (p = 0.002, R2 0.204). This study has established Porphyromonas as anumerically significant and highly prevalent genus in feline periodontal disease.     

Duration of action and hemodynamic properties of mivacurium chloride in dogs anesthetized with halothane.

OBJECTIVE: To describe onset and duration of neuromuscular blockade induced by mivacurium chloride and its associated hemodynamic effects at 3 dosages in healthy dogs. ANIMALS: 7 Labrador Retrievers. PROCEDURE: Anesthesia was induced with thiopental and maintained with halothane in oxygen, and dogs were mechanically ventilated to end-tidal P(CO)2 between 35 and 40 mm Hg. Core temperature, end-tidal P(CO)2, and halothane concentration were kept constant throughout the experiment. Neuromuscular function was assessed by evaluation of the train-of-four response to a supramaximal electrical stimulus of 2 Hz applied to the ulnar nerve every 10 seconds. Blood for determination of plasma cholinesterase activity was obtained prior to administration of mivacurium, a bolus of which was administered IV, using a randomized Latin-square design for dosages of 0.01, 0.02, and 0.05 mg/kg of body weight. RESULTS: All dogs had typical plasma cholinesterase activity. After administration of mivacurium, differences were not evident between groups in heart rate, systolic, mean, or diastolic blood pressure, change at any time in heart rate, systolic, mean, or diastolic blood pressure, or pH. Interval from onset to 100% neuromuscular blockade was 3.92+/-1.70, 2.42+/-0.53, and 1.63+/-0.25 minutes at dosages of 0.01, 0.02, and 0.05 mg/kg, respectively. Duration of measurable neuromuscular blockade was 33.72+/-12.73, 65.38+/-12.82, and 151.0+/-38.50 minutes, respectively. Time of onset and duration of effect differed significantly among dosages. CONCLUSIONS AND CLINICAL RELEVANCE: Mivacurium provides good hemodynamic stability at the dosages tested. In dogs, this drug has a rapid onset and long duration of effect.     

Microsatellite variation between an African and five European taurine breeds results in a geographical phylogenetic tree with a bison outgroup

Genetic variation is being used extensively for individual identification and linkage analysis, and may be useful for interpopulation studies. Previously, blood groups and biochemical variants in blood cell and serum proteins have been used to study (evolutionary) relationships in mammals. But genetic divergence and gene flow among closely related populations are difficult to measure with these classical markers because their mutation rate is so low that new mutations have not had sufficient time to appear and become fixed. So they have a small number of alleles and a relatively low level of heterozygosity. These markers are now replaced by DNA markers, mostly microsatellites. These microsatellite loci are useful genetic markers at which alleles differ in length due to differences in the number of short sequence motifs arranged adjacent to one another. They are abundantly distributed throughout the mammalian genome. They have a large number of alleles, a high level of heterozygosity and are inherited in true Mendelian fashion. These characteristics make them valuable for parentage control, linkage analysis, genome mapping and phylogenetic studies. In terrestrial vertebrates with limited mobility, genetic differentiation often increases with the distance between populations or corresponds to the extent of geographic and habitat barriers (R oy et al. 1994). Investigations of short tandem repeats yield a considerable volume of genetic data regarding the similarities and divergence times of different cattle populations. Microsatellite markers are suitable for the estimation of these parameters as they are not generally subject to direct selection and environmental influences. Computation of genetic distances based on data from several loci can be used to evaluate the taxonomic relationship between populations. The aim of this study was to estimate the relative genetic variability between Belgian cattle breeds and to reconstruct the evolutionary relationship among them, also using two small genetically isolated cattle-like populations.