Deleterious AGXT Missense Variant Associated with Type 1 Primary Hyperoxaluria (PH1) in Zwartbles Sheep.

Severe oxalate nephropathy has been previously reported in sheep and is mostly associated with excessive oxalate in the diet. However, a rare native Dutch breed (Zwartbles) seems to be predisposed to an inherited juvenile form of primary hyperoxaluria and no causative genetic variant has been described so far. This study aims to characterize the phenotype and genetic etiology of the inherited metabolic disease observed in several purebred Zwartbles sheep. Affected animals present with a wide range of clinical signs including condition loss, inappetence, malaise, and, occasionally, respiratory signs, as well as an apparent sudden unexpected death. Histopathology revealed widespread oxalate crystal deposition in kidneys of the cases. Whole-genome sequencing of two affected sheep identified a missense variant in the ovine AGXT gene (c.584G>A; p.Cys195Tyr). Variants in AGXT are known to cause type I primary hyperoxaluria in dogs and humans. Herein, we present evidence that the observed clinicopathological phenotype can be described as a form of ovine type I primary hyperoxaluria. This disorder is explained by a breed-specific recessively inherited pathogenic AGXT variant. Genetic testing enables selection against this fatal disorder in Zwartbles sheep as well as more precise diagnosis in animals with similar clinical phenotype. Our results have been incorporated in the Online Mendelian Inheritance in Animals (OMIA) database (OMIA 001672-9940).

Assessment of ketamine and medetomidine anaesthesia in the domestic rabbit.

OBJECTIVE: To study the effects of ketamine and two doses of medetomidine administered by two routes of injection in a genetically diverse population of rabbits. STUDY DESIGN: Prospective, randomized, clinical trial. ANIMALS: One hundred and five domestic rabbits of mixed breed, sex and age. MATERIALS AND METHODS: Rabbits undergoing orchiectomy or ovariohysterectomy received ketamine (15 mg kg(-1)) combined with medetomidine at 0.25 or 0.5 mg kg(-1), by subcutaneous (SC) or intramuscular (IM) injection. Anaesthesia was supplemented with 1.5-2% isoflurane when signs of regular jaw movements and/or slight limb twitching indicated inadequate anaesthesia. Heart and respiratory rate, blood oxygen saturation, end-tidal carbon dioxide concentration and rectal temperature were monitored at several time points. Duration of surgical anaesthesia and anaesthesia time were measured. At completion of surgery, atipamezole (1.0 or 0.5 mg kg(-1), IM or SC) was administered. STATISTICAL ANALYSES: MANOVA was used to compare variables over time between males and females, anaesthetic doses and routes of drug administration. RESULTS: All reflexes were lost significantly more rapidly after IM drug administration (p < 0.05). The times (in minutes) from drug injection to loss of reflexes for the respective groups were: righting reflex: 6.3 (15.0 + 0.25, SC), 5.5 (15.0 + 0.5, SC), 2.9 (15.0 + 0.25, IM) and 2.3 (15.0 + 0.5, IM); ear pinch: 9.2, 8.5, 4.8, 3.6; pedal withdrawal: 12.8, 10.4, 6.6, 5.2. Heart and respiratory rates during surgery did not differ between groups, however the highest end-tidal CO(2) concentration during surgery was significantly affected by dose, with the highest concentration occurring in group 15.0 + 0.5 IM. The number of animals requiring isoflurane tended to decrease with increasing dose of anaesthetic and significantly more females required supplementation than males (p < 0.05). Recovery from anaesthesia (return of righting reflex) was not significantly different between dose groups (p > 0.1) but was more rapid in animals given IM atipamezole (13.6 +/- 13 versus 21 +/- 17, p = 0.037). No anaesthetic-related mortality occurred and all but three animals recovered uneventfully. Five animals were killed whilst under anaesthesia because of unrelated disease. CONCLUSION AND CLINICAL RELEVANCE: Ketamine-medetomidine combinations reliably produced surgical anaesthesia in domestic rabbits that could easily be deepened for brief periods with low concentrations of isoflurane. Subcutaneous administration was better tolerated, but the speed of induction was slower compared with IM injection. Atipamezole was an effective antagonist and produced most rapid effects when administered IM.

Nitric oxide during perfusion improves posttransplantation function of non- heart-beating donor lungs.

BACKGROUND: We attempted to determine in a pig model whether 20 ppm of nitric oxide (NO) during perfusion ameliorates warm ischemic lung injury in the non-heart-beating donor (NHBD), thereby improving function with longer warm ischemia. METHODS: Lungs were retrieved from three groups (n=6): 1 hr (NHBD(1)) and 2 hr with and without NO (NHBD(2)NO, NHBD(2)) after hypoxic death. For assessment and preservation, left lungs were ventilated with 100% oxygen (NHBD(2)NO with added NO) and perfused for 20 min with neutrophil-depleted, deoxygenated blood in Perfadex solution. Pulmonary vascular and airway pressures and blood flow were measured with pulmonary venous blood gases. Perfusion temperature was reduced to 18 degrees C prior to storage at 4 degrees C before transplantation. RESULTS: NO during perfusion significantly improved posttransplantation pulmonary venous oxygenation (NHBD(1) [mean +/- SD] 51+/-14 kPa, NHBD(2) 54+/-16 kPa, and NHBD(2)NO 61+/-6 kPa; P=0.01) and airway pressures (NHBD(1) 30.8+/-3.5, NHBD(2) 32.5+/-5.6, NHDB(2)NO 29.4+/-5.3; P=0.0001). NO significantly improved pulmonary vascular resistance (excluding the initial cold-induced vasoconstricted reperfusion period): NHBD(1) 19+/-9 Wood units, NHBD(2) 28+/-25 Wood units, NHDB(2)NO 16+/-10 Wood units, P=0.029. Neutrophil uptake was significantly lowered by NO: NHBD(1) 0.6+/-1.4*10(9) minute-1, NHBD(2) 1.2+/-1.0*10(9) minute-1, NHBD(2)NO 0.4+/-0.9*10(9) minute-1 (P=0.029). CONCLUSIONS: This technique satisfactorily assesses and preserves the non-heart-beating lung. NO during preservation reverses the slight deterioration seen when increasing warm ischemia from 1 to 2 hr, significantly improving transplant oxygenation, vascular resistance, and airway pressures. This may be a result of the observed significant reduction in neutrophil sequestration.

Renal scarring caused by vesicoureteric reflux and urinary infection: a study in pigs.

The diminishing risk of acute renal scarring with urine infections (reflux nephropathy) after infancy is unexplained, but might reflect kidney maturation. The mechanisms of reflux nephropathy scarring are best explained by a piglet model in which vesicoureteric reflux allows infected urine to enter those segments of renal parenchyme that are drained by compound papillae. We carried out a similar study in adult pigs to determine whether protective maturation occurs. Adult pigs were exposed to urine infection after surgery to produce unilateral vesicoureteric reflux. The intravesical portion of one ureter was deroofed in six female adult Gottingen mini-pigs and the bladder and the ureteric mucosae stitched around the perimeter of the new orifice. One week later Escherichia coli was injected into the urinary bladder to produce cystitis. Three weeks later the animals were killed humanely and the urinary tracts were examined. The animals sustained persistent urine infections; the untreated ureters and kidneys remained normal. However, on the operated side, the ureters were thickened and dilated, vesicoureteric reflux was shown in four cases, and the kidneys had one or more flattened area overlying a renal segment, which showed severe inflammatory changes and early scar formation. The risk of reflux nephropathy scarring is not eliminated by maturation of the kidney in pigs. It is unlikely that the much-reduced risk of initiating scarring that is seen in older children with urine infections is due to a protective maturation of the human kidney. A possible explanation is that most children born with risk factors for developing scarring will have already sustained scars when very young.