亚洲璃眼蜱唾液腺一新功能基因的克隆与测序

HaB1是亚洲璃眼蜱雌成蜱唾液腺差异表达基因文库中的一个片段,根据其序列设计引物HaB1-GSP1和HaB1-GSP2,以唾液腺总RNA为模板,RACE法扩增获得HaB1的未知3′-末端。测定该末端序列,进行序列拼接,设计全长引物5-′CCAGTCCGAAGGAAGGGCG-3′和5-′TCTC-CGGGCACGTGAAGTGTC-3′。以cDNA第一链为模板,扩增获得基因全长。经核查表明,该基因为一新基因(登录号AY803896)。     

Canine GM2‐Gangliosidosis Sandhoff Disease Associated with a 3‐Base Pair Deletion in the HEXB Gene

Background

GM2‐gangliosidosis is a fatal neurodegenerative lysosomal storage disease (LSD) caused by deficiency of either β‐hexosaminidase A (Hex‐A) and β‐hexosaminidase B (Hex‐B) together, or the GM2 activator protein. Clinical signs can be variable and are not pathognomonic for the specific, causal deficiency.

Objectives

To characterize the phenotype and genotype of GM2‐gangliosidosis disease in an affected dog.

Animals

One affected Shiba Inu and a clinically healthy dog.

Methods

Clinical and neurologic evaluation, brain magnetic resonance imaging (MRI), assays of lysosomal enzyme activities, and sequencing of all coding regions of HEXA, HEXB, and GM2A genes.

Results

A 14‐month‐old, female Shiba Inu presented with clinical signs resembling GM2‐gangliosidosis in humans and GM1‐gangliosidosis in the Shiba Inu. Magnetic resonance imaging (MRI) of the dog's brain indicated neurodegenerative disease, and evaluation of cerebrospinal fluid (CSF) identified storage granules in leukocytes. Lysosomal enzyme assays of plasma and leukocytes showed deficiencies of Hex‐A and Hex‐B activities in both tissues. Genetic analysis identified a homozygous, 3‐base pair deletion in the HEXB gene (c.618‐620delCCT).

Conclusions and Clinical Importance

Clinical, biochemical, and molecular features are characterized in a Shiba Inu with GM2‐gangliosidosis. The deletion of 3 adjacent base pairs in HEXB predicts the loss of a leucine residue at amino acid position 207 (p.Leu207del) supporting the hypothesis that GM2‐gangliosidosis seen in this dog is the Sandhoff type. Because GM1‐gangliosidosis also exists in this breed with almost identical clinical signs, genetic testing for both GM1‐ and GM2‐gangliosidosis should be considered to make a definitive diagnosis.     

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.     

耕作方式和秸秆覆盖对旱地麦豆轮作下小麦籽粒产量、蛋白质含量和土壤硝态氮残留的影响

为明确耕作方式和秸秆覆盖对旱地麦豆轮作下小麦籽粒产量、蛋白质含量和土壤硝态氮残留的影响,2014年10月-2016年6月,利用设置在豫西典型旱作区的麦豆轮作栽培模式长期定位试验,选取传统翻耕、翻耕覆盖、旋耕和旋耕覆盖4个处理,比较了小麦氮素吸收利用、籽粒产量、蛋白质含量以及土壤硝态氮残留量。结果表明,与翻耕相比,旋耕不影响小麦产量,但花后氮素积累量、籽粒蛋白质含量和蛋白质产量分别降低60.0%、8.6%和13.0%,而成熟期0~200 cm土层硝态氮残留量显著提高28.6%。同一耕作方式下,秸秆覆盖较无覆盖不仅显著提高了穗数、穗粒数、千粒重和收获指数,还提高了拔节前和开花后的氮素积累量,促进了营养器官氮素向籽粒中转运,从而使翻耕覆盖的籽粒产量、氮素吸收效率、籽粒蛋白质含量和蛋白质产量较翻耕分别提高11.5%、13.5%、7.4%和21.3%,旋耕覆盖较旋耕也分别提高23.0%、39.5%、12.8%和38.5%。在试验进行7年后的小麦成熟期,翻耕覆盖0~200 cm土层硝态氮残留量较翻耕降低31.3%,旋耕覆盖较旋耕也降低51.4%。因此,秸秆覆盖不仅可提高旱地麦豆轮作下小麦产量、蛋白质含量和氮素吸收效率,还能降低土壤硝态氮的残留量,是兼顾旱地小麦高产优质和环境友好生产的有效途径,尤其以旋耕覆盖效果突出。     

氨基酸与母猪营养

母猪饲养是养猪生产中最重要的环节之一。母猪的营养尤其是蛋白质和氨基酸营养不仅直接影响仔猪的生长性能,而且也对母猪的使用年限和繁殖性能会产生很大影响。母猪蛋白质及氨基酸营养受品种、胎次、母猪体况、产仔数、泌乳量、采食量、哺乳期长短等诸多因素影响,增大了母猪氨基酸营养的研究难度。同时母猪个体较大、研究成本高、试验群体偏小、数据采集困难以及试验误差较大等问题也限制了母猪蛋白质及氨基酸营养的系统研究。本文对母猪的氨基酸营养进行综述,旨在为养猪生产实践中母猪日粮的合理配制提供参考。     

Evaluation of vegetable protein in canine diets: Assessment of performance and apparent ileal amino acid digestibility using a broiler model

Recent technological advances in the human food industry with respect to meat processing have decreased the availability of animal proteins to the pet food industry which typically formulates diets with an excess of animal protein. In the long term, this is not sustainable; thus, alternative protein sources need to be investigated. This study examined three canine diets, comparing a typical animal protein‐based diet (control) with two experimental diets where the animal protein was substituted in part with vegetable protein (formulated based either on total protein or amino acid content) using a broiler model. Each diet was fed to six cages each containing two birds from day 15, 18 cages in total (36 birds). Excreta were collected from days 19 to 21. On day 23, birds were euthanized and weighed, and their ileal digesta were collected and pooled for each cage. In addition, one leg per cage was collected for evaluation of muscle mass. Results showed no significant difference in animal performance (feed intake or live weight gain) or muscle to leg proportion across the diets. Birds fed the control diet and the diet balanced for amino acid content exhibited the greatest coefficients of apparent metabolizability for nitrogen (p < .001). Birds fed the diets that contained partial replacement of animal with vegetable protein generally had greater ileal digestibility of amino acids compared to birds fed the control (animal protein) diet. Analysis of excreta showed no dietary difference in terms of dry matter content; however, birds fed the diet balanced for total protein and the diet balanced for amino acid content had significantly greater excreta nitrogen than the control (p = .038). Overall, the study suggests vegetable proteins when formulated based on amino acid content are a viable alternative to animal proteins in canine diets.     

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.     

Evaluating the nutritive profile of three insect meals and their effects to replace soya bean in broiler diet

This study was conducted to evaluate the comparative effect of maggot meal, silkworm meal and mealworm as dietary protein source on the production performance and some aspects of meat quality in broilers. In this regard, maggot meal was reared on chicken offal and poultry waste. Silkworm meal was obtained from silk industry, while mealworm was developed through beetles rearing. A total of 120‐day‐old broiler chicks were randomly divided into four groups where soya bean meal (M0) was replaced with maggot meal (M1), silkworm meal (M2) and mealworm (M3) respectively. Each group was further divided into three replicates. The study was carried out for a period of 5 weeks. Diets containing mealworm significantly reduced overall feed consumption and resulted into higher weight gain (p < .05). Lowest feed conversion ratio (FCR) was recorded for birds fed with mealworm diet (p < .05). Tenderness and juiciness of meat were higher (p < .05) in M3 compared to the control and other treatments. Mortality did not vary between the control and the treated groups. Therefore, it is concluded that insect meal is rich in essential nutrients and could be successfully used in broiler ration without compromising acceptability. In the light of this study, mealworm is the best choice in broiler ration, in comparison with maggot and silkworm.