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.     

Effects of three short-term pasture allocation methods on milk production,methane emission and grazing behaviour by dairy cows

Two short-term grazing experiments were conducted with Norwegian Red cows. In Exp 1, 24 cows were randomly assigned to one of the following three pasture allocation methods (PAM): weekly pasture allowance (7RG), grazing 1/7 of 7RG each day (1SG), or grazing as 1SG but had access to grazed part of the paddock within one week (1FG). In Exp 2, 7RG was shortened to 5 days (5RG). We hypothesized that PAM will affect sward quality, quantity, intake and production differently. Pasture chemical composition changed with advancing grazing days but were not different between treatments. Pasture intake, milk yield, and methane emission were not affected by PAM. In Exp 1, 7RG cows spent less time on grazing, whereas in Exp 2, 1FG cows spent longer on grazing than others. Patterns observed in sward quality, and behavioural and physiological adaptations of cows to short-term changes in nutrient supply may explain the observed effects.     

Reliably colonising broiler chickens with Campylobacter spp. using a litter-based method

ABSTRACT

1. Chicken-associated Campylobacter spp. are the cause of most food poisoning cases in Europe. In order to study the host–pathogen interactions, a reliable and reproducible method of colonising chickens with the bacteria is required.

2. This study aimed to identify a more appropriate and less invasive method of colonisation (cf. gavaging) by seeding bedding material (litter) that commercial chickens are kept on with a mixture of Campylobacter spp., broth and faeces.

3. The first phase of the study tested the longevity of Campylobacter spp. recovery in seeded litter over 24 h: significantly more Campylobacter spp. was recovered at 0 or 3 h post-seeding than at 6 and 24 h post-seeding, indicating that the pathogen can survive to detectable levels for at least 3 h in this environment.

4. In the second phase, three groups of 10 broiler chickens (negative for Campylobacter spp. prior to exposure) were exposed at 21 days of age to one of three different Campylobacter jejuni and C. coli mixes (A, B, C), using the method above. At 28 days of age, birds were euthanised by overdose of barbiturate or cervical dislocation, and livers and caeca removed for Campylobacter spp. assessment.

5. All liver and 28/30 caeca samples tested positive for Campylobacter spp., with mix A and C giving higher counts in the caeca than mix B. The method of euthanasia did not affect Campylobacter spp. counts.

6. In conclusion, a successful method for reliably colonising broiler chickens with Campylobacter spp. has been developed which negates the need for gavaging and is more representative of how contamination occurs in the field.     

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets.

(2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial.

(3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle).

(4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.     

Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.