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.     

断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响

为了研究断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响,选取24只体况良好、胎次一致、体重相近的10日龄羔羊(湖羊),将其随机分为3组,在饲喂相同母乳的基础上,分别补饲以纯木薯(CS)、玉米(MS)和豌豆淀粉(PS)(直/支链淀粉比分别约为0.11,0.27和0.44)为唯一淀粉来源的开食料。试验期间,每天4:00-19:00将羔羊抱入补饲栏补饲不同直/支链淀粉比的开食料,并且在6:30, 10:30和15:30将羔羊抱回母羊舍哺乳1 h。羔羊自由饮水,单栏饲喂,自由采食燕麦干草。56日龄时屠宰采样,采集瘤胃组织样品制作石蜡切片进行瘤胃乳头形态测定,采集瘤胃上皮样品提取RNA测定相关基因表达。结果表明:CS组羔羊瘤胃乳头长度和表面积显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃乳头宽度显著(P=0.001)高于PS组。对瘤胃上皮各层厚度统计显示,CS和PS组羔羊瘤胃上皮角质层厚度显著(P=0.001)高于MS组;CS和MS组羔羊瘤胃上皮颗粒层厚度显著高于(P<0.001)PS组;CS组羔羊瘤胃上皮棘基层厚度和总厚度显著(P<0.001)高于MS和PS组。qRT-PCR结果显示:不同直/支链淀粉比开食料显著影响羔羊瘤胃上皮CDK2和CDK6的mRNA表达量(CS>MS>PS, P<0.001);CS组羔羊瘤胃上皮细胞cyclin A和CDK4的mRNA表达量显著(P<0.05)高于PS组,但与MS组无显著差异;CS组羔羊瘤胃上皮cyclin D1的mRNA表达量显著(P=0.012)低于PS组,但与MS组无显著差异。CS组羔羊瘤胃上皮类胰岛素生长因子-1(insulin like growth factor,IGF-1)的mRNA表达量显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃上皮IGF-1R的mRNA表达量显著(P=0.001)高于PS组。上述结果表明:较高支链淀粉比开食料与较高直链淀粉比开食料相比,在瘤胃中较易降解生成挥发性脂肪酸,促进瘤胃上皮IGF-1及细胞周期蛋白mRNA的表达,进而有利于断奶前羔羊瘤胃上皮的发育。研究结果对制订羔羊营养方案具有重要指导意义。     

中国荷斯坦牛部分乳房结构特性对泌乳性能的影响

本研究对规模化场奶牛乳房结构部分性状进行评价和测定,旨在分析其影响因素及其与泌乳性能的关系。现场评价和测定我国北方6个规模化奶牛场荷斯坦牛的乳房均衡(2 030条)、乳头长度(1 551条)和乳头位置评分(分前、后乳头,2 839条),使用SAS 9.2 GLM过程采用固定效应模型分析影响上述3个乳房性状的因素,同时分析3个单项性状对校正日产奶量和体细胞评分的影响。结果表明:场和胎次对3个乳房性状影响极显著(P0.01);前、后乳头位置和后乳头长度对校正日产奶量有显著影响(P0.05),校正日产奶量与前乳头位置评分值呈正比,与后乳头位置评分值呈反比;前乳头位置和长度对体细胞评分有显著影响(P0.05),体细胞评分随前乳头位置评分的增大而减小;5~5.5 cm的乳头长度对于校正日产奶量和体细胞评分都最佳。本研究结果为通过乳房性状选择、淘汰并提高奶牛的生产性能、乳房健康提供了参考。     

A comparison of the effect of short-acting and long-acting cloxacillin-based dry-cow therapy on somatic cell counts after calving in cows also given internal teat sealants

AIM: To compare, in cows treated with an internal teat sealant, the effect of short-acting and long-acting cloxacillin-based dry-cow therapy on somatic cell counts (SCC) after calving.

METHODS: Cows from a spring-calving, pasture-based dairy farm in the Manawatu-Whanganui region of New Zealand were randomly allocated to receive either a short-acting cloxacillin and ampicillin dry-cow therapy and internal teat sealant (n=291) or a long-acting cloxacillin and ampicillin dry-cow therapy and internal teat sealant (n=288) at the end of lactation. Cows were managed on-farm with routine husbandry procedures through the dry period and following calving. A multivariable logistic regression model was used to determine the association between length of action of dry-cow therapy and the proportion of cows with a SCC >150,000?cells/mL at the first herd test after calving.

RESULTS: Age of cow, mean SCC for the preceding season and interval from calving to the first post-calving herd test were all associated with the proportion of cows with an individual SCC >150,000?cells/mL at the first herd test (p<0.001) Treatment with the short-acting dry-cow therapy was not associated with decreased odds of cows having a SCC >150,000?cells/mL at the first herd test compared with treatment with long-acting dry-cow therapy (OR=0.724; 95% CI=0.40–1.30).

CONCLUSIONS AND CLINICAL RELEVANCE: In this herd, which routinely used internal teat sealants, the use of short-acting cloxacillin-based dry-cow therapy did not result in an increased proportion of cows with elevated SSC post-calving. This was a single farm, single year study but indicates that in this herd, changing from a long-acting to a short-acting antimicrobial may have no impact on the prevalence of subclinical mastitis.     

Repeat patient testing shows promise as a quality control method for veterinary hematology testing

Background

Repeat patient testing‐based quality control (RPT‐QC) is a potential method for veterinary laboratories (eg, that have a limited budget for quality commercial control material [QCM] or that wish to use material with a species‐specific matrix).

Objectives

To determine whether total error (TE_a), probability of error detection (Ped), and probability of false rejection (Pfr) similar to that achievable with QC materials can be controlled using RPT‐QC

Methods

Control limits (WBC, RBC, HGB, HCT, MCV, and PLT) for the Advia 120 (n = 23) and scil Vet ABC (n = 22) were calculated using data from normal canine specimens from a routine caseload. Specimens were measured at accession and again after 24 hours. Control limits were validated using 23 additional canine specimens tested similarly. Achievable TEa, Ped, and Pfr were investigated using the Westgard EZRules3 and compared to those achievable with commercial QCM.

Results

Theoretical performance of RPT‐QC and commercial QCM‐QC are similar for 1‐3s with both n = 1 and 1‐3s with n = 2 for all measurands and both instruments. Achievable TE_a values for RPT‐QC were close to ASVCP recommendations for most measurands; exceptions were PLT (both instruments) and WBC (scil Vet ABC).

Conclusions

Repeat patient testing‐based quality control advantages include a species‐specific matrix, low‐cost, and absence of QC material deterioration over time (since a fresh specimen is used each day). A potential disadvantage is daily access to normal canine specimens. A challenge is determining control limits, which has a subjective element. Further study is needed to confirm actual RPT‐QC performance and to determine if RPT‐QC with abnormal patient specimens is feasible.     

Interaction between the sequence of feeding of hay and concentrate,and boiling of barley on feed intake,the activity of hydrolytic enzymes and fermentation in the hindgut of Arabian mares

The interaction between the sequence of feeding of hay and concentrate and the hydrothermal processing of barley in alleviating concentrate effects on intake, and hindgut fermentation in horses was tested. Six Arabian mares (4–10 years of age, 410 ± 35 kg body weight) were used to evaluate the effects of feeding sequence (FS) and type of barley (TB) on intake, and faecal volatile fatty acids (VFA), activities of α‐amylase (AA: EC 3.2.1.1), carboxymethyl cellulase (CMCase: EC 3.2.1.4), microcrystalline cellulase (MCCase: EC 3.2.1.91) and general filter paper degrading activity (FPD). Mares were offered a ration of air‐dried alfalfa and concentrate (70:30 as‐fed) in four subsequent periods of 14 days including 8 days of adaptation and 6 days of sampling. In each period and each meal, mares received concentrate either 30 min after (HC) or 30 min before (CH) alfalfa hay. Barley was either milled or boiled in water. Rectal samples were grabbed directly from rectum once per period. Mares subjected to CH had higher dry matter intakes than mares under HC regime. The acetate:propionate ratio (A:P ratio) in rectal content was higher with CH than HC. The AA activity was higher under CH than under HC. Mares fed boiled barley had lower rectal concentrations of VFA and propionate and a higher A:P ratio than mares fed milled barley. Furthermore, the rectal content showed a higher MCCase activity but a lower AA activity when mares were fed boiled compared with milled barley. Interactions between FS and TB were observed with respect to CMCase activity, and concentrations of propionate and valerate. In conclusion, the present results suggest that both, feeding concentrate before hay and boiling the barley, might improve the hindgut environment in Arabian mares, and that the two measures were mostly additive and sometimes even synergistic.     

Effect of bovine seminal plasma on bovine endometrial epithelial cells in culture

The purpose of this study was to investigate the effect of seminal plasma (SP) from bulls of known fertility on bovine endometrial epithelial cells (bEEC) in culture. The bEEC from passage 5, approximately 5.0–13 × 10⁵ cells per flask, were challenged with SP from bulls of high or low fertility (n = 3 and 2, respectively) or PBS (control), at 1% (75 μl) or 4% (300 μl) and were incubated for 72 hr (n = 13 per challenge). Total cell number and viability of bEEC after challenge with 1% SP from either high‐ or low‐fertility bulls (75H or 75L, respectively) did not differ from controls. In contrast, challenge with 4% of SP from high‐ or low‐fertility bulls (300H or 300L) negatively affected bEEC cell number and viability. Challenge with 300 L had a greater adverse effect than 300H. These results suggest that the negative effect of bovine SP on bEEC is both dose‐dependent and fertility‐dependent.