Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.     

Assessing the effectiveness of manure export plus intensive silage cropping for lowering the Olsen‐P status of P‐enriched grassland

A substantial proportion of farmed grassland soils in Northern Ireland (NI) are overly enriched with P and pose a risk to water quality. To address this problem, manure could be exported rather than recycled to P‐enriched land and the latter intensively cropped with grass silage to deplete soil P. To assess the efficacy of such a strategy, a P‐ and K‐enriched grassland site was intensively cropped over a 6‐yr period with fertilizer N alone supplied to support silage growth. By year 6, soil P had declined from index 5 to index 3, and it was estimated that two more years of this management may bring it into the target index 2 range. Soil K, however, declined rapidly from index 4 to index 1 in just 4 yr, with the result that grass production became limited by K deficiency. It was concluded that nonrecycling of manure to P‐enriched grassland under silage management is probably the most effective strategy for lowering soil P status, but care must be taken to prevent K deficiency occurring.     

Predicting the degree of phosphorus saturation using the ammonium acetate–EDTA soil test

As a result of the important role played by phosphorus (P) in surface water eutrophication, the susceptibility of soils to release P requires evaluation. The degree of phosphorus saturation, assessed by oxalate extraction (DPS_ox), has been used as an indicator. However, most laboratories do not include DPS_ox in routine soil tests because of cost and time. This study evaluates the suitability of the ammonium acetate extraction in the presence of EDTA (AAEDTA), the standard soil test P (STP) in Wallonia (Southern Belgium), to predict DPS_ox; we also compared it with the Mehlich 3 extraction. Ninety‐three topsoil samples were collected in agricultural soils throughout Wallonia. Good correlations were found between the AAEDTA and the Mehlich 3 methods for P, Fe and Al (r = 0.85, 0.77 and 0.86, respectively). An exponential relationship was found between P_AAEDTA and DPS_ox. Results of principal component analysis and regression demonstrated that STP can be used to predict DPS_ox (r = 0.93) after logarithmic transformation. Soil test Al was also a good indicator of the P sorption capacity (PSC_ox) of soils (r = 0.86). Including the clay fraction in regression equations only slightly improved the prediction of PSC_ox (r = 0.90), while other readily available data (such as pH or organic carbon) did not significantly improve either DPS_ox or PSC_ox predictions.     

Critical comparison of the vanadomolybdate and the molybdenum blue methods for the analysis of phosphate in plant sap

Abstract

The performance of the vanodomolybdate and molybdenum blue methods for the routine determination of phosphate in plant sap were compared in order to assess their suitability for the detection of P deficiency at an early stage of plant growth. The molybdenum blue method was ca 10 times more sensitive than the vanadomolybdate one and required less sap to give a successful test. Recoveries of phosphate added to both sap and solution samples were close to 100% with both methods. The methods were in good agreement over a wide range of phosphate concentrations using sap samples from all leaves during the early stages of P deficiency, but increases in sap background colour caused the vanadomolybdate method to overestimate phosphate concentrations as the deficiency became more acute. Measurements on solutions of similar absorbance showed that the vanadomolybdate method was not significantly more variable than the molybdenum blue method but tended to overestimate phosphate concentration when very small volumes of sap were available. It was concluded that the molybdenum blue method was the most reliable for routine measurements of phosphate in plant sap.     

Genetics of equine bleeding disorders

Genetic bleeding disorders can have a profound impact on a horse's health and athletic career. As such, it is important to understand the mechanisms of these diseases and how they are diagnosed. These diseases include haemophilia A, von Willebrand disease, prekallikrein deficiency, Glanzmann's Thrombasthenia and Atypical Equine Thrombasthenia. Exercise-induced pulmonary haemorrhage also has a proposed genetic component. Genetic mutations have been identified for haemophilia A and Glanzmann's Thrombasthenia in the horse. Mutations are known for von Willebrand disease and prekallikrein deficiency in other species. In the absence of genetic tests, bleeding disorders are typically diagnosed by measuring platelet function, von Willebrand factor, and other coagulation protein levels and activities. For autosomal recessive diseases, genetic testing can prevent the breeding of two carriers.     

Hepatic lipidosis: Liver characteristics and acute phase proteins in affected turkeys

The hepatic lipidosis (HL) in fattening turkeys is a disease has been known for a long time, but the cause and pathogenesis is still not clarified. A recent study reported unexplained high levels of iron in liver tissue of fattening turkeys suffering from HL. In this study, the iron status, possible infectious or inflammatory influences in form of an acute phase reaction and the analysis of fatty acid pattern in liver tissue of turkeys affected by HL were examined. Three cases of HL on three different fattening turkey farms were investigated during the outbreak of the disease. Clinically affected and non-affected animals were subjected to a pathological examination, where the diagnosis HL or non-affected was made. In total, 70 birds were examined (40 with HL, 30 without HL) and blood and liver samples were taken. Additionally, samples from 15 slaughtered birds were taken as a further control group. In liver tissue, the iron content and the content of long-chain fatty acids were determined; in blood samples, ferritin and transferrin were measured. The iron content in liver tissue was more than three times higher for animals with HL than among non-affected animals and the control group. The transferrin levels were lowest for animals with HL, highest in the control group and in between for non-affected animals. The fatty acid pattern in liver tissue of affected animals indicated a shift from polyunsaturated fatty acids to saturated fatty acids and monounsaturated fatty acids compared to the control group and the non-affected animals. Overall, the non-affected animals of a flock affected by HL were similar to the healthy animals of the abattoir. The low acute phase protein levels for animals with HL together with high iron contents could indicate a previous malnutrition/starvation period and/or severe liver damage for those animals suffering from HL.     

Mesh-like vascular changes in copper deficiency-induced rat cardiomyopathy

The pathological effects of copper deficiency (COD) are well known. However, the pathogenesis of cardiomyopathy resulting from COD remains unclear. In this study, aimed to elucidate the pathogenesis of COD-induced cardiomyopathy by examining the morphology of the cardiovascular system in copper-deficient rats using histopathology, immunohistochemistry, and scanning and transmission electron microscopy. Changes detected in the myocardium and interstitium were consistent with those reported for COD. Morphological changes included mesh-like changes in the capillary endothelial cells that appear to be a novel finding in COD-induced cardiomyopathy. These changes are hypothesized to result from abnormal vascular remodeling following damage to the basement membrane and due to the mechanical effects of myocardial contractions. Although cardiomyopathy may be associated with microcirculatory disorders arising from these lesions, further investigations are necessary to demonstrate a causal relationship between the pathogenesis of cardiomyopathy and the contribution of these lesions to disease progression.     

Relationship between a plant-based ‘vegan’ pet food and clinical manifestation of multiple nutrient deficiencies in two cats

A topical subject in human nutrition is the steadily growing number of people choosing to limit or completely avoid all animal-derived food products either for moral dilemma, health concerns or both. To meet people's will of applying their dietary choices to their domestic animals, the pet food industry answered by launching on the market some plant-based diets. This leads to concerns about whether these diets are adequately formulated to satisfy the target species nutritional requirements, especially for cats which are still considered strict carnivores. This case report follows a 2-year-old male neutered Main Coon and a 1-year-old female spayed Domestic Shorthair cat, presented to the nutrition service of the University of Toulouse, France. Reason for consultation was lethargy with in anamnesis a recent dietary transition to a plant-based pet food. Dysorexia, lethargy and muscle waste were present at first consultation. Progressive weight loss developed during follow-ups. A macrocytic, non-regenerative anaemia with serum folates below reference were the main clinical features. Analysis of pet food showed multiple nutrients below minimum recommendation at the average daily intake of both cats. Folic acid supplementation improved dysorexia, and subsequent reintroduction of animal-derived ingredients in the diet restored appetite, weight and a normal mentation in both cases.     

灰铸铁生产技术的一些新进展

对灰铸铁的研究现状和生产中的一些新技术做了评述。重点介绍了高强度薄壁灰铸铁的生产现状以及先进的制造工艺和先进的制造手段在灰铸铁生产中应用所取得的技术成果。     

Mineral absorption in the black rhinoceros (Diceros bicornis) as compared with the domestic horse

To test whether mineral recommendations for horses are likely to guarantee adequate mineral provision for black rhinoceroses (Diceros bicornis), we investigated the apparent absorption (aA) of macro- and microminerals in eight black rhinoceroses from three zoological institutions in a total of 32 feeding trials with total faecal collection, with additional data from three unpublished studies (18 feeding trials). Feeds and faeces were analysed for Ca, P, Mg, Na, K, Fe, Mn, Cu, Zn and Co. The resulting aA coefficients, and the linear relationships of apparently absorbable dietary mineral content to total dietary mineral content [per 100 g dry matter (DM)], were compared with data for domestic horses. Rhinoceroses had significantly higher aA coefficients for Ca and Mg (because of a higher calculated 'true' absorption), and lower ones for Na and K (because of calculated higher endogenous faecal losses). High absorption efficiency for divalent cations is hypothesized to be an adaptation to a natural diet of particularly high Ca:P ratio (approximately 14:1); an effective removal of Ca from the ingesta guarantees sufficient P availability at the fermentation site in the hindgut. Higher faecal losses of Na and K are hypothesized to be linked to a higher faecal bulk per DM intake in black rhinoceroses as compared with horses because of a generally lower digestive efficiency. There were no relevant differences in the absorption patterns of microminerals. In particular, there were no discernable differences in Fe absorption within the rhinoceroses for diets with and without tannin supplementation. Several of the zoo diets assessed in this study were deficient in Cu, Mn or Zn, and most contained excessive levels of Fe when compared with horse requirements. The findings of this study indicate that differences in mineral absorption between occur even between species of similar digestive anatomy; that in particular, Ca absorption might vary between hindgut fermenters with Ca:P ratio in their natural diet; that Na might be a particularly limiting factor in the ecology of free-ranging rhinoceroses; that moderate doses of tannins do not seem to markedly influence mineral absorption; and that diets for captive animals should contain adequate, but not excessive mineral levels.