Analysis of the effect of the bovine adenosine triphosphate-binding cassette transporter G2 single nucleotide polymorphism Y581S on transcellular transport of veterinary drugs using new cell culture models

In commercial dairy production, the risk of drug residues and environmental pollutants in milk from ruminants has become an outstanding problem. One of the main determinants of active drug secretion into milk is the ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP). It is located in several organs associated with drug absorption, metabolism, and excretion, and its expression is highly induced during lactation in the mammary gland of ruminants, mice, and humans. As a consequence, potential contamination of milk could expose suckling infants to xenotoxins. In cows, a SNP for this protein affecting quality and quantity of milk production has been described previously (Y581S). In this study, our main purpose was to determine whether this polymorphism has an effect on transcellular transport of veterinary drugs because this could alter substrate pharmacokinetics and milk residues. We stably expressed the wild-type bovine ABCG2 and the Y581S variant in Madin-Darby canine kidney epithelial cells (MDCKII) and MEF3.8 cell lines generating cell models in which the functionality of the bovine transporter could be addressed. Functional studies confirmed the greater functional activity in mitoxantrone accumulation assays for the Y581S variant with a greater relative V(MAX) value (P = 0.040) and showed for the first time that the Y581S variant presents greater transcellular transport of the model ABCG2 substrate nitrofurantoin (P = 0.024) and of 3 veterinary antibiotics, the fluoroquinolone agents enrofloxacin (P = 0.035), danofloxacin (P = 0.001), and difloxacin (P = 0.008), identified as new substrates of the bovine ABCG2. In addition, the inhibitory effect of the macrocyclic lactone ivermectin on the activity of wild-type bovine ABCG2 and the Y581S variant was also confirmed, showing a greater inhibitory potency on the wild-type protein at all the concentrations tested (5 μM, P = 0.017; 10 μM, P = 0.001; 25 μM, P = 0.008; and 50 μM, P = 0.003). Differential transport activity depending on the genotype together with the differential inhibition pattern might have clinical consequences, including changes in substrate pharmacokinetics (and subsequently pharmacodynamics) and more specifically, changes in secretion of ABCG2 substrates into milk, potentially implying important consequences to veterinary therapeutics.     

The pharmacokinetics and metabolism of ivermectin in domestic animal species

The pharmacokinetic properties of drugs are closely related to their pharmacological efficacy. The kinetics of ivermectin are characterised, in general terms, by a slow absorption process, a broad distribution in the organism, low metabolism, and slow excretion. The kinetics vary according to the route of administration, formulation, animal species, body condition, age, and physiological status, all of which contribute to differences in drug efficacy. Characterisation of ivermectin kinetics can be used to predict and optimise the value of the parasiticide effects and to design programmes for parasite control. This article reviews the pharmacokinetics of ivermectin in several domestic animal species.     

Morphology and amplitude values of the electrocardiogram of Spanish-bred horses of different ages in the Dubois leads system

The aim of this work was to record the modifications of the form and amplitude of the electrocardiographic tracings during growth using the Dubois system and to study its sensibility to these physiological changes. This work was carried out on 179 healthy, young and adult Spanish-bred horses (98 females and 81 males). One electrocardiogram (ECG) was obtained from each horse using the Dubois leads system. The bifid shape deflection of the P wave was generally more frequent than the simple one in animals at 4 months of age or older. Amplitudes of the P2 component and of the simple positive wave presented significant differences (p < 0.01) between the age groups studied: animals up to 3 months in age and older than 2 years of age (highest amplitudes), and the rest of the animals (lowest amplitudes). Significant variations (p < 0.05) were found for the QS amplitude between animals of 1 month of age, and all the other groups. The multiple range analysis did not define a clear distribution of age groups for the amplitude of the T wave. It may therefore be concluded, that in the Spanish-bred horse up to 1 month of age, the form and amplitude of the QRS complex in the Dubois leads system differ from those of older animals. Thus, this study shows the sensibility of the Dubois leads system for detecting the electrocardiographic changes related to the growing process in the Spanish-bred horse. In addition, it provides standard values of electrocardiographic parameters in the Spanish-bred horse at different stages of growth.     

A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Endometrial Adenocarcinoma and Mucometra in a 6-year-old Alaska Malamute Dog

A 6-year-old female Alaska Malamute dog was presented for evaluation of abdominal enlargement referred by a local veterinarian. On the history, the owner complained of chronic abdominal enlargement initiated more than 4 months ago, reduced appetite, occasional vomiting and general dullness. He also complained of greenish mucous intermittent vaginal discharge starting 10 days ago. The bitch was chronically treated with medroxiprogesterone acetate. A laparatomy was performed and fluid in the abdomen was found and aspirated during the surgery. Also a very fluid-filled distended uterus and a mass in the distal part of the left uterine horn were found. The mass was encapsulated by the omentum, but areas of necrosis and calcification were identified. Histopathological diagnosis was endometrial adenocarcinoma.     

Autonomic Innervation of the Equine Urinary Bladder

The distribution and density of intrinsic autonomic nerve fibers and cells were studied in the equine urinary bladder by means of the peroxidase-antiperoxidase immunohistochemical method to localize tyrosine-hydroxylase (TH), and by means of a histochemical technique to detect acetylcholinesterase (AChE) activity. The results suggest that the equine urinary bladder, like that of other mammalian species, possesses a rich autonomic innervation which includes catecholaminergic and acetylcholinesterase positive nerves. At least a part of these nerve fibers have an intrinsic origin from ganglion cell bodies within the bladder wall.