Frequency of the mutant MDR1 allele associated with multidrug sensitivity in a sample of collies from France

A study was performed to determine the frequency of the mutant MDR1 allele associated with ivermectin sensitivity in a sample of collies living in France. Buccal swab samples were collected from approximately 83 collies for determination of MDR1 genotype. DNA was extracted and the polymerase chain reaction was performed to amplify a 148 bp (wildtype MDR1 genotype) or 144 bp (mutant MDR1 genotype) amplicon containing the MDR1 mutation. Sequence analysis was performed to determine the genotype of each dog. Adequate quantities of DNA for unequivocal genotyping were obtained from only 25 of 83 swabs. Twenty percent (5/25) of the collies studied were homozygous for the normal allele (normal), 32% (8/25) were heterozygous (carrier), and 48% (12/25) were homozygous for the mutant allele (affected). The results of this study indicate that a high percentage of collies presenting to veterinarians in France harbor the MDR1 mutation, thus impacting some therapeutic decisions.     

A direct method for fatty acid methyl ester synthesis: application to wet meat tissues, oils, and feedstuffs

A simplified protocol to obtain fatty acid methyl esters (FAME) directly from fresh tissue, oils, or feedstuffs, without prior organic solvent extraction, is presented. With this protocol, FAME synthesis is conducted in the presence of up to 33% water. Wet tissues, or other samples, are permeabilized and hydrolyzed for 1.5 h at 55 degrees C in 1 N KOH in MeOH containing C13:0 as the internal standard. The KOH is neutralized, and the FFA are methylated by H(2)SO(4) catalysis for 1.5 h at 55 degrees C. Hexane is then added to the reaction tube, which is vortex-mixed and centrifuged. The hexane is pipetted into a gas chromatography vial for subsequent gas chromatography. All reactions are conducted in a single screw-cap Pyrex tube for convenience. The method meets many criteria for fatty acid analysis, including not isomerizing CLA or introducing fatty acid artifacts. It is applicable to fresh, frozen, or lyophilized tissue samples, in addition to oils, waxes, and feedstuffs. The method saves time and effort and is economical when compared with other methods. Its unique performance, including easy sample preparation, is achieved because water is included rather than eliminated in the FAME reaction mixtures.     

Influence of body condition and bovine somatotropin on estrous behavior, reproductive performance, and concentrations of serum somatotropin and plasma fatty acids in postpartum Brahman-influenced cows

Ninety-nine multiparous Brahman-influenced (1/4 to 3/8 Brahman) cows were managed to achieve low (BCS = 4.3 +/- 0.1; n = 50) or moderate (BCS = 6.1 +/- 0.1; n = 49) body condition (BC) to determine the influence of bovine somatotropin (bST) on estrous characteristics, reproductive performance, and concentrations of serum GH and plasma NEFA. Beginning 32 d postpartum, cows within each BC were assigned randomly to treatment with or without bST. Non-bST-treated cows received no treatment, and treated cows were administered bST (Posilac, 500 mg s.c.) on d -35, -21, and -7 before initiation of the breeding season. On d -7, all cows received an intravaginal, controlled internal drug-releasing (CIDR) device. On d 0 (initiation of the 70-d breeding season), the CIDR were removed and cows received prostaglandin F2alpha (PGF2alpha). Blood samples were collected from the median caudal vein of the cows at each bST treatment and at d -28 and 0. Estrous behavior was monitored by radiotelemetry during the first 30 d of the breeding season. Growth hormone was increased (P < 0.05) in low and moderate BC cows treated with bST. The percentage of cows detected in estrus during the first 30 d of the breeding season was decreased (P = 0.05) for low BC (64%) compared with moderate BC (82%) cows. The interval to first estrus tended (P = 0.07) to be shorter in low BC-bST-treated cows (3.7 +/- 1.9 d) than in moderate BC-bST-treated cows (9.6 +/- 1.8 d). During the first 30 d of the breeding season, cows in low BC had a decreased (P = 0.02) number of mounts received and increased (P = 0.001) quiescence between mounts compared with cows in moderate BC. The number of mounts received was reduced (P = 0.04) in bST-treated cows. More (P = 0.02) cows treated with bST became pregnant during the first 3 d of the breeding season compared with non-bST-treated cows. The cumulative first-service conception rate tended (P = 0.07) to be greater for bST-low BC cows than non-bST-treated cows in low or moderate BC. On d 0, NEFA were greater (P < 0.05) in bST-treated vs. non-bST-treated cows. Low BC and bST reduced the intensity of behavioral estrus in postpartum Brahman-influenced cows. However, bST increased the first-service conception rate during the first 30 d of breeding and pregnancy rates during the first 3 d of breeding in postpartum Brahman-influenced cows.     

A spontaneous and fatal jejunal intussusception in a European brown hare associated with Eimeria leporis

A recently dead adult female European brown hare (Lepus europaeus Pallas) was found in the wild with no evidence of trauma or external injury. Necropsy revealed symptoms consistent with endotoxic shock and a haemorrhagic jejuno-jejunal intussusception with a necrotic intussusceptum. Small, unattached nodules were present extensively in the jejunal lumen and within the intussuscipiens. These nodules proved to be masses of unsporulated Eimeria leporis oocysts. Review of the literature reveals E. leporis to be a common parasite of wild hares, with protracted pathogenic potential under some circumstances. This is the first report of intussusception in this species and the most severe lesion seen associated with E. leporis. This case suggests that E. leporis may be more harmful to the wild European brown hare population than previously supposed.     

Developing a classification tool based on Bloom's taxonomy to assess the cognitive level of short essay questions

The cognitive level of short essay questions taken from assessments of two veterinary courses at the Faculty of Veterinary Medicine of Utrecht University (FVMU) was evaluated using a simplified classification tool based on the taxonomy of Bloom. Classifications were made by teaching staff members (subject matter experts, or SME) and by faculty members not involved in teaching the course (non-subject matter experts, or NSME). To compare the cognitive level assigned by raters in the SME group to that assigned by the NSME group, each test item was assigned a modal taxonomic level. The results indicate that the agreement level between a pair of raters within a group (SME or NSME) differed (34% to 77% and linear weighted Cohen's kappa coefficient 0.12 to 0.60). The agreement level on the modal taxonomic level between the SME and NSME groups for the two courses was 65% and 73%, with a linear weighted Cohen's kappa coefficient of 0.43 and 0.63 respectively. The requirement of expertise of a subject for classification is discussed. The introduction of the classification tool had a positive effect on teaching staff members' awareness of the importance of the cognitive level of assessments. Improvements to the classification tool to obtain higher agreement levels are proposed.     

Effect of anionic salt and highly fermentable carbohydrate supplementations on urine pH and on experimentally induced hypocalcaemia in cows

The objective of this experiment was to determine the effect of dietary grain on calcium homeostasis. Six rumen-fistulated dairy cows with 3 or more previous lactations and no history of parturient paresis were randomly assigned to a sequence of diets in a crossover study with 4 periods of 10 days each. Dietary treatments were: A control ration consisting of wrap grass silage alone (1), the control ration supplemented with ammonium chloride and ammonium sulphate salt solution (2), control ration following a period with supplementation (3) and control ration supplemented with increasing amounts of barley from 4 to 10 kg/cow per day, expected to produce subclinical rumen acidosis (4). Daily intake of the diets was adjusted to 14 kg DM/cow per day. On day 11, the calcium-regulating mechanisms in cows were challenged until recumbency by a standardized intravenous EDTA infusion and cows were left to recover spontaneously. Anion supplementation and the feeding of highly fermentable carbohydrate lowered urine pH below 7.0 due to subclinical acidosis. During spontaneous recovery from EDTA induced hypocalcaemia, the cows more quickly regained a whole blood free calcium concentration of 1.00 mmol/L if they had most recently been supplemented with either anionic salts or with increasing amounts of barley, as compared to the basic ration. It is concluded that so-called slug-feeding or 'steaming up' with highly fermentable carbohydrates before parturition in milk fever susceptible cows enhanced calcium homeostasis similar to the effect seen in cows on anionic diets.     

Development of an Immunobinding Assay with Monoclonal Antibodies to Diagnose Mycoplasma bovis in Semen

Veterinary Research Communications -     

Serotyping and virulence genes detection in Escherichia coli isolated from fertile and infertile eggs, dead-in-shell embryos, and chickens with yolk sac infection

Escherichia coli is a common avian pathogen mainly associated with extraintestinal infections such as yolk sac infection (YSI). The aim of this study was to determine the serotypes and the presence of some virulence genes of E. coli strains isolated from different samples in a vertically integrated poultry operation in Mexico. Two hundred sixty-seven E. coli isolates from different samples were serotyped using rabbit serum against the 175 somatic (O) and 56 flagellar (H) antigens of the typing schema. Virulence genes were determined by colony blot hybridization, using DNA probes for st, eae, agg1, agg2, bfp, lt, cdt, slt, and ipaH diarrhea-associated virulence factors. The serogroup of 85% of the strains was determined; O19 (12%), 084 (9%), 08 (6%), and 078 (5%) were the most common. Using the complete antigenic formula (O and H), O19:NM (n = 31) was the serotype most frequently isolated from dead-in-shell embryos and in broilers that had died on the fourth, fifth, sixth, and seventh days after hatch. One hundred ten strains (41.2%) hybridized with one or more of the used probes. Of these, ipaH (72%), eae (30%), and cdt (27%) were the most common. Considering the origin of the respective isolates, 40% of the broiler farm strains were positive for at least one probe. Results show that some avian E. coli strains isolated in Mexico are included in avian pathogenic E. coli serotypes not previously reported, suggesting that they could be specific for this geographic area. The wide distribution of the ipaH gene among nonmotile strains suggests that this invasiveness trait could be important in YSI pathogenesis. On the other hand, some other genes could contribute to E. coli virulence during YSI.     

Combined dorsolateral and intraoral approach for the resection of tumors of the maxilla in the dog

This paper describes in detail a combined dorsal and intraoral approach for maxillectomy for tumors involving tissues more caudal to the third premolar. The only intraoperative complication was that of blood loss, with six out of 20 dogs requiring a single unit of blood. Histopathologically clean margins were obtained in 14 of the 20 cases, with a recurrence rate of 50% in these dogs and a median time to recurrence of 24 months. This represents an improvement in outcome over previously reported studies, and the authors postulate this is due to the better exposure and access to the area afforded by the combined approach over the standard intraoral approach.     

Feline intraocular tumors may arise from transformation of lens epithelium

Feline ocular sarcomas are malignant intraocular neoplasms that are frequently associated with a history of ocular trauma. They usually present as fibrosarcomas, but some have both epithelial and mesenchymal features. The purpose of this study was to determine the cell of origin of a subset of feline intraocular sarcomas that display a mixed epithelial-mesenchymal phenotype, with elaboration of basement membrane-type matrix. We examined the morphology and histochemical and immunohistochemical phenotypes of nine feline intraocular sarcomas. Immunohistochemistry and in situ hybridization were performed to detect expression of crystallin alpha A. In addition, tumors were examined for expression of vimentin, cytokeratin, smooth muscle actin, desmin, melan A, neural cell adhesion molecule, S-100, glial fibrillary acidic protein, nerve growth factor receptor, and collagen type IV. Animals ranged from 7 to 17 years of age--no breed or sex predilection for tumor occurrence was present. Tumors were characterized by mixed epithelial and mesenchymal phenotypes, both of which elaborated basement membrane-type material and expressed vimentin highly. On the basis of collagen type IV and crystallin alpha A immunopositivity, we established that three of nine tumors were of lens epithelial origin. Expression of desmin and smooth muscle actin identified one tumor as a leiomyosarcoma. The remainder were undifferentiated sarcomas of myofibroblastic origin. This is the first report of lens epithelial neoplasia in clinical material from any species. The history and morphologic features of feline ocular sarcomas are reminiscent of feline vaccine-induced sarcomas. These tumors may share pathophysiologic similarities unique to this species.