西农萨能奶山羊SERPINA1基因克隆、生物信息学及组织表达分析

本研究旨在克隆西农萨能奶山羊SERPINA1基因的CDS区,采用生物软件和在线预测工具进行生物信息学分析,通过实时荧光定量PCR技术检测SERPINA1基因在西农萨能奶山羊各组织间mRNA的表达水平。根据GenBank中山羊SERPINA1基因CDS区序列（登录号：XM_018066209.1）,利用Primer Premier 5.0软件设计特异性引物,RT-PCR扩增目的基因,构建原核表达载体测序后对序列进行生物信息学分析;采集西农萨能奶山羊心脏、肝脏、脾脏、肺脏、乳腺、肾脏、肌肉、瘤胃和小肠组织,提取组织RNA,反转录为cDNA模板,设计特异性定量引物,进行实时荧光定量PCR,检测SERPINA1基因在不同组织中的表达差异。结果显示,西农萨能奶山羊SERPINA1基因CDS区全长1 326 bp,编码441个氨基酸;同源性比对分析显示,西农萨能奶山羊与山羊、绵羊、牛和小鼠SERPINA1基因核苷酸序列同源性分别为100%、98.6%、95.7%和71.6%,与山羊亲缘关系最近,其次是绵羊。SERPINA1蛋白分子质量为48.71 ku,等电点为5.71,为跨膜亲水蛋白;SERPINA1氨基酸序列分别有62个磷酸化位点,3个跨膜区结构。组织表达分析显示,SERPINA1基因在西农萨能奶山羊肝脏组织中显著高表达（P<0.05）,其次是乳腺组织,在肺脏组织中表达量最低。研究结果为进一步探究SERPINA1基因在奶山羊乳蛋白合成代谢中的作用提供理论依据。     

Evaluation of dietary stevioside supplementation on anti-human serum albumin immunoglobulin G, Alpha-1-glycoprotein, body weight and thyroid hormones in broiler chickens

Sixty male broiler chickens fed a diet supplemented with 130 mg/kg stevioside (S group) or an unsupplemented diet (C group) from day 1 of age onwards. On day 21 of age, ten birds from either the S (SH) or C (CH) group were injected subcutaneously with 100 μg human serum albumin (HSA) and ten others from either S (SP) or C (CP) group injected with 100 μl phosphate-buffered saline (PBS) in the same way. There were no significant effect of supplementation nor interaction with age on average body weights, T(3) and T(4) concentrations of non-injected chickens. After the primary immunization, α(1) -glycoprotein concentrations increased in all treatment groups except the CP group, and were significantly higher in the CH group in relation to the other groups. Fourteen and 18 days after the primary immunization, HSA injected chickens of both dietary treatments had significantly higher anti-HSA immunoglobulin G (IgG) levels than their PBS injected controls. No effect of stevioside supplementation was observed for IgG level. In conclusion, dietary stevioside inclusion can attenuate the pro-inflammatory response after stimulation of the innate immune response in broiler chickens.     

Immunohistochemical localization of substance P,calcitonin gene-related peptide,galanin and calcium-binding proteins in trigeminal ganglia of goat (Capra hircus)

The objectives of this study was to provide a quantitative analysis of calcium-binding proteins, calbindin (CB), parvalbumin (PA), substance P (SP), calcitonin gene-related peptide (CGRP) and galanin (GAL), in trigeminal ganglia of goats, to establish whether they exhibit coexistence relationships between each other, and to examine possible colocalization with SP, CGRP and GAL, which have been well characterized according to their distributions in an abundance of large and/or small neurones. CB (12.78%), PA (31.91%), SP (24.63%), CGRP (44.44%) and GAL (3.29%) immunoreactive (IR) cells were observed. About 38.37, 8.7 and 0.73% of CGRP-IR neurones in the trigeminal ganglion were also immunoreacted with SP, GAL and CB, respectively. Almost all SP-IR cells are labelled with CGRP (approximately 92.52%), whereas only 16.02 and 0.44% of SP-IR neurones colocalized with GAL and CB. Approximately 4.65 and 1.10% of the CB-IR cells were found to contain CGRP and SP immunoreactivity, respectively. Conversely, no CB-IR cell exhibited GAL immunoreactivity. In addition, all the GAL-IR cells showed CGRP and SP immunoreactivity. The number of CB-, PA-, SP-, CGRP- and GAL-IR neurones in goat trigeminal ganglion are abundant than that of other animals. These results elucidate that the goat differs from other mammalian species in the distribution and localization of neurochemical substances in trigeminal ganglia, and suggest that this difference may be relevant to the morphological characteristics of cerebral vasculatures such as epidural rete mirabile of goat.     

Effects of dietary supplementation of modified zinc oxide on growth performance,nutrient digestibility,blood profiles,fecal microbial shedding and fecal score in weanling pigs

One hundred and forty piglets ((Landrace × Yorkshire) × Duroc, 21 day of age) with an initial weight of 6.50 ± 0.71 kg, were randomly allotted into four treatments to determine the effects of a modified form of zinc oxide (ZnO) on growth performance, nutrient digestibility, blood profiles, fecal microbial shedding and fecal score in weanling pigs. Dietary treatments were: (i) NC, negative control, basal diet containing zinc (Zn) from the premix; (ii) PC, positive control, basal diet containing Zn‐free premix + 3000 ppm ZnO; (iii) H1, basal diet containing Zn‐free premix + 3000 ppm ZnO (phase 1, days 1 to 14)/200 ppm modified ZnO (phase 2, days 15 to 42); (iv) H2, basal diet containing Zn‐free premix + 300 ppm modified ZnO (phase 1)/200 ppm modified ZnO (phase 2). During days 1 to 14, average daily gains (ADG) were higher (P = 0.04) in PC, H1 and H2 groups than that in NC group. Overall, H1 treatment increased the ADG compared with NC (P = 0.05). On day 14, the alkaline phosphatase and plasma Zn concentration were increased (P = 0.01 and 0.04, respectively) in PC, H1 and H2 treatments compared with NC treatment. On days 14 and 42, the fecal Lactobacillus counts in NC group were lowest (P = 0.01, P = 0.04 respectively) among treatments. All supplemented groups showed lower (P = 0.03) fecal score than NC treatment on days 21 and 28. In conclusion, dietary supplementation with modified ZnO increased growth rates and reduced fecal scores in weanling pig. Modified ZnO could be used as a substitute to ZnO as a growth promoter and reduce Zn excretion to the environment because of the lower dosage. [Correction added on 3 February 2015, after first online publication: the initial weight of ‘6.50 ± 1.11 kg’ has been replaced with ‘6.50 ± 0.71 kg’ in the abstract.]     

山羊PPARα基因克隆、分子特性及组织差异表达分析

试验旨在克隆山羊过氧化物酶体增殖物激活受体α（peroxisome proliferators-activated receptors-alpha,PPARα）基因的CDS区序列,分析其编码蛋白的结构与功能,并探讨其在山羊不同组织中的表达模式。采用RT-PCR方法扩增并克隆PPARα基因CDS编码区;通过在线软件对其一级结构、二级结构和三级结构进行生物信息学分析;利用基因序列和编码蛋白构建系统发育树,进行系统发育进化分析;采用实时荧光定量PCR方法检测PPARα基因在简州大耳羊心脏、肺脏、肝脏、肾脏、脾脏和网膜6个组织中的相对表达情况。结果表明,山羊PPARα基因CDS区全长1 413 bp,结构稳定,共编码470个氨基酸。生物信息学分析发现,PPARα蛋白是一种结构较为稳定的带负电的亲水性蛋白,以α-螺旋和无规则卷曲为主,无信号肽和跨膜蛋白,属于膜内蛋白。蛋白序列中总共有49个磷酸化位点,9个糖基化位点。保守结构域中含有明显的DBD区域和LBD区域,蛋白结构高度保守。三级结构预测发现其蛋白结构主要为通过长链卷曲连接的2个明显不同的结构区域,结构均以helix螺旋为主。序列比对结果表明,山羊PPARα氨基酸序列与绵羊和牛的同源性最高。实时荧光定量检测结果表明,PPARα基因在肾脏和肝脏中表达量较高,显著高于其他组织（P < 0.05）;在网膜组织和心脏中中度表达,显著高于肺脏和脾脏（P < 0.05）;在肺脏和脾脏中相对低表达;说明山羊PPARα基因可能与体内脂肪氧化、脂质代谢和抗氧化应激等调控功能有关。试验结果为深入研究PPARα基因在山羊中的生理功能和调控机制奠定了理论基础。     

Investigations on the light and scanning electron microscopic structure of the lingual papillae in the angora goat (Capra hircus): II. Mechanical papillae

This study was conducted to examine the light microscopy (LM) and scanning electron microscopic (SEM) structures of mechanical papillae on the tongue in the Angora goat (Capra hircus). As study materials, the tongues of four adult female Angora goats were used. The samples were collected from the dorsal surfaces of the apex, body, root and torus of the tongue and the ventral surfaces of the lingual apex for light and scanning electron microscopic examinations. Three types of mechanical papillae were seen in Angora goat tongue: papilla filiformes, papilla lentiformes and papilla conicae.The filiform papillae were detected in the dorsal surface of the tongue from lingual apex to lingual torus, and in the ventro-lateral of the lingual apex and on both sides of the lingual torus. The morphological differences were observed in filiform papillae according to their location in the tongue. The lenticular papillae settled on the centre of the lingual torus. Two types of these papillae, irregular-round and pyramid-shaped were identified. The conical papillae were scattered all over the lingual torus, except for the central part and were also seen on the root of the tongue. In the light microscopic examination, it was found that mechanical papillae had a stratified squamous epithelium and a varying degree of keratin layer on epithelial surfaces. In this study, by examining the light and scanning electron microscopic structure of the mechanical papillae in the Angora goat tongue, their similarities and differences with other domestic and wild ruminant species were determined.     

谷氨酸锌对肉仔鸡生长性能、血清含锌酶活性及免疫器官指数的影响

本试验旨在研究谷氨酸螯合锌(zinc glutamine,Glu-Zn)对肉仔鸡生长性能、血清含锌酶活性、免疫器官指数的影响。选用240只7日龄商品代AA肉用仔鸡,随机分成6个处理。1组为对照组饲喂含80mgZnSO4·H2O形式的Zn的基础日粮,其余5组分别在基础日粮中添加20、40、60、80mg/kg和100mg/kg谷氨酸螯合锌形式的Zn。饲养期为35d。结果表明:试验各周及全期,随着日粮中谷氨酸螯合锌水平的增加,肉仔鸡日增重和饲料转化效率均得到改善,同时,40mg/kg谷氨酸螯合锌组显著提高了试验全期的平均日增重(P0.05);血清碱性磷酸酶活性随着日粮锌水平的增加先上升,而后又下降,在锌添加水平为60mg/kg时取得最大值;谷氨酸螯合锌组在胸腺指数、脾脏指数上,明显优于对照组,且存在显著差异(P0.05),而法氏囊指数并不敏感。从本试验各项指标和经济效益综合分析,建议肉仔鸡饲料中添加60mg/kg的谷氨酸螯合锌。     

Effects of field bean (Vicia faba L. var. minor) dietary supplementation on plasma thyroid hormones, insulin, insulin-like growth factor-1 concentrations and mohair characteristics in growing Angora goat kids

Twenty-two Angora kids were used to study the effect of a dietary supplementation with field bean (Vicia faba L. var. minor) on hair follicle activity, mohair characteristics and plasma total thyroxine (T4), triiodothyronine (T3), insulin (INS) and insulin-like growth factor-1 (IGF-1) concentrations. At birth, their mothers were divided in two groups (S: supplemented, C: control), the S group being supplemented with 300 g/head/day of whole field bean. At weaning, diet of kids from S group was supplemented with 80 g/head/day of whole field bean. At weaning, secondary hair follicle activity (S: 0.91 ± 0.01, C: 0.84 ± 0.02, p < 0.01) and staple length (S: 5.82 ± 0.13 cm, C: 5.16 ± 0.14 cm, p < 0.001) were greater in the S group. At 155 days of age, secondary fibre diameter (S: 16.54 ± 0.35 μm, C: 18.09 ± 0.31 μm, p < 0.01) was higher in the C kids. Concentrations of total plasma T4 and T3 were higher (p < 0.001) in S (120 ± 12 and 4.87 ± 0.92 ng/ml respectively) than in C kids (92 ± 8 and 2.97 ± 0.77 ng/ml respectively). At weaning, T4 values were negatively correlated (p < 0.05) with the secondary fibre diameter and the T3/T4 ratio was positively correlated (p < 0.05) with the secondary to primary hair follicle ratio. At 155 days of age, both T3 and T3/T4 values were positively correlated (p < 0.01) with staple length. Plasma INS (0.11 ± 0.03 ng/ml) and total IGF-1 concentrations (232 ± 13 ng/ml) were not affected by diet. Improvements of diet quality in kids positively affected fibre diameter and length and follicle activity rate. We suggest that increased circulating thyroid hormones concentrations, but not those of INS and IGF-1, may contribute to the beneficial actions of nutritional supplementation on anatomical growth and mohair production of kids.     

葛藤草粉替代苜蓿草粉对波杂山羊血清生化指标、养分表观消化率和生产性能的影响

为了解葛藤草粉在山羊全混合日粮中等量替代紫花苜蓿草粉的效果,探索葛藤草粉在当地山羊生产中的应用价值,首先对葛藤草粉的常规营养成分进行测定,然后选择体重[(18.20±1.65)kg]相近的5月龄波尔山羊×马头山羊阉公羔75只,随机分为5组,每组15只.对照组饲喂基础日粮,其余4个处理组分别以葛藤草粉等量替代紫花苜蓿草粉...     

Concentrations of thyroid hormones and iodothyronine binding proteins in serum of the koala (Phascolarctos cinereus)

Objective To examine circulating total and free thyroid hormone (T₃ and T₄) concentrations, determine serum iodothyronine binding characteristics and estimate thyroid stimulating hormone (TSH) activity in sera of coastal and inland koalas.
Design A prospective study.
Procedure Koala serum T₃ and T₄ were measured by radioimmunoassay. T₄ binding parameters were determined by radioligand binding and electrophoresis. Koala TSH values were determined by bioassay.
Results Mean total T₄ concentrations were 3.2 ± 2.1 nM although values were significantly higher in inland-dwelling females in comparison to coastal-dwelling males. Free T₄ was 3.3 ± 2.1 pM. Total and free T₃ were 0.4 ± 0.2 nM and 1.4 ± 0.9 pM respectively, although these values were at the lower end of the assay detection limit and should be viewed with reservation. Electrophoresis of [¹²⁵I]-T₄-labelled serum revealed only two proteins of electrophoretic mobility similar to human transthyretin (TTR) and albumin. Scatchard analysis of T₄ binding to serum gave a curvilinear plot, which could be resolved into two binding sites with affinities identical to that of TTR and albumin but both of low concentration. The bioactivity of the TSH present in the sera was measured using a cell line (JP09) transfected with the human TSH receptor. The mean level of stimulation found in the sera corresponded to a bovine TSH activity of less than 10 mU/L.
Conclusion These results suggest that the serum concentrations of free and total thyroid hormones in koalas are low compared to other marsupials and very low compared to eutherian mammals. The mechanism of maintenance of euthyroidism in this species remains to be determined.     

Effect of age on serum chemistry profile, electrophoresis and thyroid hormones in beagle dogs two weeks to one year of age

Chemistry tests were conducted on serum from young Beagle dogs in order to deter mine the effect of age on these parameters. Blood was collected at regular intervals from 25 normal Beagle puppies (13 males and 12 females) at ages ranging from 2 weeks to 12 months. Serum chemistry profiles, protein electrophoresis and radioimmunoassays for thyroxine and triiodothyronine were included. Rapidly changing age related differences were observed in several parameters. Urea nitrogen, cholesterol, triglycerides, lactate dehydrogenase, thyroxine, glucose, gamma glutamyl transpeptidase, and total bilirubin values were elevated early in life, and decreased during the first 6 to 8 weeks, while alanine aminotransferase activity was low initially and increased during this period. Lactate dehydrogenase, thyroxine, gamma glutamyl transpeptidase, total bilirubin and alanine aminotransferase attained stability by 3 months, but the remaining parameters showed slight changes subsequently, gradually approaching adult values. More gradual age related changes were observed in other parameters. These included alkaline phosphatase, inorganic phosphorus and calcium values, which were higher in younger dogs, and creatinine, aspartate aminotransferase and total protein values, which were lower in younger dogs. Creatinine and aspartate aminotransferase values were stable by approximately 6 months; alkaline phosphatase, inorganic phosphorus, calcium and total protein values continued to change gradually up to 1 year.     

The expressions of some metabolic hormones (leptin,ghrelin and obestatin) in the tissues of sheep tongue

In this study, we aimed to observe the localization and expression of peptide hormones—leptin, ghrelin and obestatin—in the sheep tongue by immunohistochemistry. For that purpose, tongues of ten adult sheep were used. Leptin expression of moderate intensity was observed in the basal and parabasal epithelial cells of the luminal epithelium, and leptin was strongly expressed in the taste buds of the circumvallate and fungiform papillae and in von Ebner's glands. Ghrelin was primarily expressed in some of the skeletal muscle cells and the smooth muscle cells of the middle layer of blood vessels. A strong expression was observed in the epithelial cells lining the base of the groove surrounding the circumvallate papillae. Obestatin expression was particularly strong in the epithelial cells of the salivary ducts. It was also stronger in the von Ebner's glands than in the seromucous glands. Leptin, ghrelin and obestatin were shown to be produced at varying levels in different cell types, including epithelial, stromal and skeletal muscle cells, as well as in ganglion neurons, neural plexuses and blood vessels in the sheep tongue. Cellular localization and expression of these peptide hormones have not been investigated in many species including sheep.     

Dietary supplementation of potassium sorbate,hydrated sodium calcium almuniosilicate and methionine enhances growth,antioxidant status and immunity in growing rabbits exposed to aflatoxin B1 in the diet

The present experiment was conducted to evaluate the effect of dietary supplementation with some feed additives (potassium sorbate; Sor, hydrated sodium calcium almuniosilicate; Hsc and L-methionine; L-M) against aflatoxin B1 (AF) toxicity in rabbits. A total of 72 growing rabbits (5-week-old) were distributed into six equal groups (4 replicates with 3 rabbits each). The experimental groups are as follows: control group, AF group (supplemented with AF 0.3 mg/kg diet), AF + Sor group (AF 0.3 mg/kg diet + Sor 2 g/kg diet), AF + Hsc group (AF 0.3 mg/kg diet + Hsc 5 g/kg diet), AF + L-M group (AF 0.3 mg/kg diet + L-M 8 g/kg diet) and AF + Mix group (AF 0.3 mg/kg diet + 2 Sor + 5 Hsc + 8 L-M g/kg diet). Live body weight and weight gain at 13 weeks of age were significantly reduced by AF. Feed intake at 13 weeks of age was decreased in AF, AF + Hsc and AF + Mix compared to the control. AF, AF + Hsc and AF + Mix showed the lowest total antioxidant capacity compared to the control. The highest level of reactive oxygen species and 8-Hydroxy-2-desoxyguanosine was observed in AF group. Using of other supplements with AF increased immunoglobulinM than AF alone. In conclusion, dietary supplementation of Sor, L-M, Hsc or their mixture was effective in reducing the adverse effects of AF on performance, antioxidant and immune status of rabbits with more better improvement obtained by Sor or L-M separately.     

Post-vaccination antibodies profile against Peste des petits ruminants (PPR) virus in sheep and goats of Punjab,Pakistan

A total of 70 sheep and 330 goats were selected randomly. All the animals were kept under same housing and management conditions. Serum samples were collected from all the animals and tested for the presence of antibodies against Peste des petits ruminants (PPR) virus using competitive ELISA (cELISA). All the animals were found negative showing percentage inhibition (PI) values <50. The animals were vaccinated against PPR with Nig/75/1 strain vaccine of PPR Serum samples were collected from randomly selected 12 sheep and 30 goats at 10, 30 and 45 days post-vaccination. The samples were subjected to cELISA to determine the presence of antibodies against PPRV. The samples with PI >50 were considered as sero-positive. The sheep found positive at 10, 30 and 45 days post-vaccination were 1(8.3%), 7(58.3%) and 12(100%) respectively. In case of goats 3(10.0%), 29(96.6%) and 27(90.0%) animals gave positive results at 10, 30 and 45 days post-vaccination respectively. Mean PI values in sheep at 10, 30 and 45 days post-vaccination were recorded as 37, 65 and 91 respectively, whereas in goats these values were 43, 78 and 86 respectively.     

Effect of dietary saponin rich soapnut (Sapindus mukorossi) shell powder on growth performance,immunity, serum biochemistry and gut health of broiler chickens

This study was conducted to evaluate the effects of dietary soapnut (Sapindus mukorossi) shell powder (SSP), a cheap source of saponins, on growth performance, immunity, serum biochemistry and gut health of broiler chickens. The experimental design was 4×2, employing four saponin levels (0, 100, 150 and 200 mg/kg diet), each provided for two time durations (0–42 day and 21–42 day) resulting into eight dietary treatments. Results revealed no significant effect of dietary saponins on body weight gain, feed intake and feed conversion ratio of birds. The abdominal fat percentage, heterophil to lymphocyte ratio, serum cholesterol and triglyceride levels, faecal total plate count, coliform count and E. coli count decreased (p < .05) progressively with increasing saponin levels and lower values were observed at 150 mg and 200 mg saponin levels. Significant improvement of cell‐mediated and humoral immune response was observed in birds fed 150 mg and 200 mg saponin compared to control. The serum glucose concentration was significantly (p < .05) higher in control group compared to other groups. No significant effects of dietary saponin were observed on carcass characteristics, faecal Lactobacillus count, intestinal histomorphometry and cost economics of broiler chicken production. Thus, dietary saponins at 150 mg/kg diet as SSP for three weeks (21–42 days) was optimum for better immunity and welfare of birds without adverse effects on the growth performance.     

吡啶甲酸铬对吉富罗非鱼生长、饲料利用和非特异性免疫功能的影响

本试验研究饲料不同水平吡啶甲酸铬对其生长、饲料利用,体成分、血液生化指标和非特异性免疫功能的影响。600尾吉富罗非鱼分为5个处理组,每个处理各设4个重复,每重复30尾鱼。每个处理铬添加量分别为0、0.4、0.8、1.2 mg/kg和10.0 mg/kg,在0.38 m3玻璃纤维钢桶中用开放式流水养殖,饲养8周。结果表明,饲料中添加吡啶甲酸铬对吉富罗非鱼的增重率、特异生长率、饲料系数、蛋白效率及蛋白质沉积率均有显著影响(P<0.05)。当铬添加量为0～0.8 mg/kg时,随着添加量增加,可显著提高吉富罗非鱼增重率、特定生长率、蛋白质效率和蛋白质沉积率(P<0.05),显著降低饲料系数(P<0.05),但对成活率和肝体比无显著影响(P>0.05);当饲料中添加为1.2 mg/kg～10.0 mg/kg时并未能进一步提高罗非鱼的生长和饲料利用。同时,铬添加水平为0～0.8 mg/kg,随着添加水平增加,显著提高吉富罗非鱼全鱼的蛋白质和磷含量(P<0.05),显著降低全鱼水分、脂肪和灰分含量(P<0.05),并显著提高吉富罗非鱼血清总蛋白含量以及肝胰脏AKP、ACP和LZM的活性(P<0.05),显著降低血清葡萄糖含量(P<0.05),但对血清胆固醇和甘油三酯含量及肝胰脏T-SOD活性无显著影响(P>0.05)。当饲料中添加铬1.2 mg/kg和10.0 mg/kg,除了显著降低肝胰脏AKP活性外(P<0.05),对血清生化指标和肝胰脏非特异性免疫酶活性无显著影响(P>0.05)。综合生长、饲料利用、体组成成分、血液生化指标和肝胰脏非特异性免疫酶活性的影响结果表明,对于0.91～4.93 g吉富罗非鱼,饲料中以吡啶甲酸铬的形式添加0.8～1.2 mg/kg铬为宜。