Fecal microbiota of three bactrian camels(Camelus ferus and Camelus bactrianus) in China by high throughput sequencing of the V3-V4 region of the 16S rRNA gene

This study aimed to reveal the microbial diversity in the fecal samples of bactrian camels using the 16 S r RNA sequencing analysis on the Illumina Mi Seq platform. Three fecal samples were collected from two geographical regions in China. Operational taxonomic unit（OTU） clustering was performed by identifying an OTU at 97% sequence identity. The alpha and beta diversities were applied to estimate the differences in microbial diversity among the three fecal samples. Totally, 4409, 3151 and 4075 OTUs in the fecal samples were identified in the Lop Nor wild camel（Camelus ferus）, the domestic camel（C. bactrianus） and Dunhuang wild camel（C. ferus）, respectively. The majority of bactreria were affiliated with phylum Firmicutes and Bacteroidetes in the three samples. The wild camels had higher gastrointestinal tract microbial diversity than the domestic one, while the microbial composition of the Lop Nor wild camel shared higher similarity with domestic camel at the genus and family levels than that of the Dunhuang wild camel did. Our results may provide a theoretical basis for assessing their health conditions and may thus be useful for protecting the critically endangered species of C. ferus.

Aetiology and Pathogenesis Studies on Osteoporosis in Bactrian Camel

A disease characterized by emaciation, pica, lameness and liability to fracture in bactrian camel from gravel desert pasture was described. Analyses of mineral elements in soil, water and forage from the affected and normal areas as well as in blood, hair and parts of tissues from normal and diseased camel, together with a pathological study were carried out to define the nature and major causes of the disease. The related blood indices were also measured. The result indicated that copper and phosphorus in the soil and forage from the affected area were significantly lower than those of the normal area (P<0.01). The levels of phosphorus in the blood, hair and rib reduced significantly in affected camels. The concentrations of PTH, T3, T4, creat-inine(Crt) and the activities of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) in serum rose markedly (P<0.01). Bone injury was characterized by osteoporosis. The degenerative and necrotic lesions of liver and kidney were common. In addition, slight demylination of brains and spinal cords were showed by histopathological and ultrastructural studies. It is concluded that the disease is caused mainly by phosphorus deficiency in the food chain.

Pre-evaluation of humoral immune response of Bactrian camels by the quantification of Th2 cytokines using real-time PCR

With the increasing immunological studies on camels due to the advantage of their single-chain antibodies for humanizations,it is demanding to develop an easy-to-handle evaluation method of their humoral immune response before proceeding with immunization of foreign antigens that may be toxic to camels.In this study,we quantitatively determined the expression levels of T-helper 2(Th2) cytokines in peripheral blood lymphocytes obtained from Bactrian camels by real-time PCR.The recorded kinetic profiles resulting from the immunization of ovalbumin(OVA) indicated that after immunization,Th2 cytokines including interleukin(IL) families such as IL-4,IL-10,and IL-13 in the camels were up-regulated by a factor of 1.78,3.15,and 1.22,respectively,which was validated by traditional enzyme-linked immunosorbent assay(ELISA) methods.Unlike ELISA which requires specific enzyme-labeled antibodies,this established method based on the minimal amount of blood samples holds an advantage in the preliminary evaluation of camel humoral immune response with desirable precision,which is meaningful for biomedical explorations of camel-derived antibodies.

Macroanatomy of Basal Nuclei in Bactrian Camel (Camelus bactrianus)

The purpose of this study was to describe the basal nuclei of brain in Bactrian camel. Seven coronal slices of brain were uniformly taken between callosal genu and splenium, and six uniform sagittal slices of left cerebral hemisphere of brain were examined. The results demonstrated that the basal nuclei of the brain include the caudata nucleus, putamen, putamen and pallidum, internal capsule and external capsule, claustrum and amygdaloid body. The study provides an elementary finding on the gross anatomy for further research on the comparative neuroanatomy, neu- rephysiology and the related behavior study of the camel.

16S rRNA Gene Sequence Analysis of Snow Leopard, Gray Wolf, Horse and Bactrian Camel in Mongolia

In this study, mitochondrial 16S rRNA sequences of snow leopard, gray wolf, domestic horse and Bactrian camel inhabited or domesticated in Mongolian territory were obtained by using polymerase chain reaction （PCR） based on universal primers for 16S rRNA （F-5＇-AACGAGCCTGGTGATA-3＇ and R-5＇-CTCCGGTCTGAACTCAGATCACGTA-3＇）. The 16S rRNA sequence was 1,048 bp to 1,086 bp in length, and each sequence was compared to other related species （Felidae, Camelidae, Equidae and Canidae） by using NCBI Basic Local Alignment Search Tool （BLAST）. Results showed that sequences were highly similar to sequences in GenBank database （93%-99%）. Then phylogenetic analysis was performed based on about 1,100 bp sequence of 16S rRNA for Panthera uncia, Canis lupus, Equus caballus, Camelus bactrianus and other related species. The result revealed that P. uncia and P. leo were sister species, C. bactrianus and C. ferus were more closely related species, and wolf and dog were the almost similar species. This finding could be important for designing species specific primers for PCR based analysis of animal species identification and forensic veterinary medicine.

Characterization of heavy-chain antibody gene repertoires in Bactrian camels

Camelids are the only mammals that can produce functional heavy-chain antibodies(HCAbs).Although HCAbs were discovered over 30 years ago,the antibody gene repertoire of Bactrian camels remains largely underexplored.To characterize the diversity of variable genes of HCAbs(VHHs),germline and rearranged VHH repertoires are constructed.Phylogenetics analysis shows that all camelid VHH genes are derived from a common ancestor and the nucleotide diversity of VHHs is similar across all camelid species.While species-specific hallmark sites are identified,the non-canonical cysteines specific to VHHs are distinct in Bactrian camels and dromedaries compared with alpacas.Though low divergence at the germline repertoire between wild and domestic Bactrian camels,higher expression of VHHs is observed in some wild Bactrian camels than that of domestic ones.This study not only adds our understanding of VHH repertoire diversity across camelids,but also provides useful resources for HCAb engineering.

A comprehensive analysis of immunoglobulin heavy chain genes in the Bactrian camel(Camelus bactrianus)

Heavy chain only antibodies(HCAbs)represent a rare type of antibody that is devoid of light chains and the CH1 domain that have been reported in cartilaginous fish and camelids.By analyzing transcript data and genome sequences,we conducted a comprehensive analysis of Bactrian camel immunoglobulin heavy chain genes.Based on the transcript data,oneμgene,fiveγgenes,oneαgene and oneεgene were found.Additionally,the variable region of HCAbs(VHH)and the conventional antibodies(VH)sequences associated with theγ3,γ1a/b andμgenes were amplified.Based on these genome sequences,seven DH,six JH,μ,γ2a,γ2c,α,andεgenes and a portion of aγ3 gene were observed.Different Kozak sequences within different VH families were found in our analysis,and the variability index differed between the VHH3 and VH3 families.Phylogenetic analysis of the constant regions of the camelid immunoglobulin genes indicates that these genes appeared before the evolutionary divergence of Bactrian camels and dromedaries.

高糖培养的双峰驼肾细胞抗氧化基因检测

为了揭示双峰驼肾脏在高糖环境中的抗氧化能力,以其离体培养的肾皮质和髓质细胞为材料,以双峰驼生理血糖值7.1 mmol/L浓度的葡萄糖培养基为对照,10、15、25和40 mmol/L葡萄糖浓度进行高糖逆境模拟培养,检测各处理组的活性氧(ROS)水平和抗氧化基因的水平。结果显示:高糖应激下,肾皮质细胞内ROS含量在10、15、25 mmol/L糖浓度下有极显著增加(P<0.01),肾髓质细胞内ROS含量在15、25、40 mmol/L糖浓度下有极显著增加(P<0.01);肾皮质细胞中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)2、SOD3、谷胱甘肽过氧化物酶(Gpx)1、Gpx3、溶质载体家族6成员1(SLC6A1)、溶质载体家族14成员1(SLC14A1)在10 mmol/L糖浓度下mRNA表达量极显著上升(P<0.01);Gpx1、Gpx3、Gpx4、糖皮质激素受体(GR)、过氧化物酶体生物发生因子5(PEX5L)、多聚结合蛋白2(PCBP2)、ATP柠檬酸裂合酶(ACLY)在40 mmol/L糖浓度下mRNA表达量极显著上升(P<0.01)。肾髓质细胞中CAT、Gpx1、Gpx3在10 mmol/L糖浓度下mRNA表达量极显著上升(P<0.01);CAT、SOD1、Gpx1、Gpx3在40 mmol/L糖浓度下mRNA表达量极显著上升(P<0.01);SLC6A1在25 mmol/L糖浓度下mRNA表达量极显著上升(P<0.01)。提示:双峰驼肾皮质细胞对糖浓度变化敏感,糖浓度偏离生理范围即引发ROS增加,激活抗氧化基因表达,产生显著氧化应激反应;相比之下,肾髓质细胞对糖浓度变化较不敏感,仅在极端糖浓度偏离时表现出类似反应。高糖环境下两者均展现抗氧化能力,但肾皮质细胞表现出更强的抗氧化适应性。

从品种特性到功能基因组:双峰驼基因组学研究进展

双峰驼是亚洲中部戈壁沙漠地区重要的畜种之一,具有耐寒、耐热、耐饥饿、免疫系统较强等独特的生物学特性,受到国内外学者的广泛关注。近年来,随着生物测序技术的快速发展,双峰驼全基因组测序及其功能基因组的研究相继发布。该文归纳了不同家养双峰驼品种特性,总结了双峰驼全基因组研究现状,回顾了双峰驼起源驯化历程,并重点介绍了双峰驼沙漠适应能力和免疫相关功能基因的研究进展,为全面了解双峰驼独特的生物学特性和遗传改良提供理论参考。

基于UPLC-MS非靶向代谢组学分析不同年龄双峰驼肉代谢物差异

基于超高效液相色谱-质谱联用的非靶向代谢组学方法探究不同年龄组驼肉代谢物的差异及变化规律。结果表明,在3个年龄组(3~4、6~7岁和9~10岁,分别以I、II和III组表示)骆驼背最长肌中共鉴定出显著差异代谢物710种;在I vs II组显著差异代谢物有78个,其中I组上调47个,II组上调31个;在II vs III组显著差异代谢物有49个,其中II组上调18个,III组上调31个;在I vs III组显著差异代谢物有65个,其中I组上调29个,III组上调36个。京都基因与基因组百科全书通路分析结果表明,差异代谢物主要富集到蛋白质和氨基酸代谢、脂肪酸、维生素和矿物质代谢等相关通路,说明不同生长阶段骆驼中各营养素的消化代谢均有差异。I组中的多不饱和脂肪酸(polyunsaturated fatty acid,PUFA)(尤其n-6 PUFA、n-3 PUFA)含量和PUFA/不饱和脂肪酸值均显著高于III组,这主要与相关代谢通路上的花生四烯酸、亚油酸和13-L-过氧化氢油酸浓度的显著上调有关;同时L-亮氨酸、L-缬氨酸、L-谷氨酰胺等差异代谢物可以作为不同年龄驼肉品质差异的潜在标记物。

双峰驼感染魏氏梭菌与溶血性曼氏杆菌的防治

2022年5月以来,甘肃省某养驼场的部分驼羔先后出现阵咳、呼吸急促、步态摇摆等症状,个别伴有腹泻,并在短期内连续死亡。经现场调查及临床检查后,初步怀疑为细菌性病原感染所致。为探明病因并控制病情,现场采集发病驼羔血清及鼻拭子进行实验室检测;同时对病死驼羔进行解剖观察,无菌采集内脏组织及肠内容物进行实验室检测。剖检发现,病死驼羔胃肠道及肺脏组织病变较为明显;镜检可见革兰阳性菌和革兰阴性菌;实验室检测发现,魏氏梭菌和溶血性曼氏杆菌核酸呈阳性。表明驼羔发病是由于感染了魏氏梭菌和溶血性曼氏杆菌引起的。通过采取紧急免疫、隔离治疗、消毒灭源等综合措施后,驼羔病情逐步好转,养驼场恢复了正常生产。

不同乳产量、乳蛋白率双峰驼血液生化指标和激素水平的对比研究

为了研究不同乳产量和乳蛋白率泌乳双峰驼血液生化指标和激素含量的差异,选用24峰体况、胎次相近的处于泌乳初期的阿拉善双峰驼,分为高产乳量组、低产乳量组、高乳蛋白组、低乳蛋白组,测定其血液指标,包括血清生化指标和血清激素水平。结果显示,在乳产量分组中,高产组双峰驼钙离子和钾离子显著低于低产组(P<0.05),非酯化脂肪酸极显著低于低产组(P<0.01),血清胰岛素浓度显著高于低产组(P<0.05);在乳蛋白率分组中,高乳蛋白组的血尿素氮浓度极显著高于低乳蛋白组(P<0.01),高乳蛋白组的孕酮和雌三醇显著低于低乳蛋白组(P<0.05),催乳素、雌酮和瘦素极显著低于低乳蛋白组(P<0.01)。结果表明,在相同的饲养管理下,离子代谢和糖代谢对乳产量的差异起到重要作用,乳蛋白的合成效率的差异可能主要由激素调节。

柯坪县规模化养殖场双峰驼隐孢子虫感染情况调查与种类鉴定

[目的]掌握新疆维吾尔自治区阿克苏地区柯坪县规模化养殖场双峰驼隐孢子虫的感染情况和种类分布情况。[方法]从柯坪县4个乡(镇)6个规模化双峰驼养殖场共收集516份粪便样本,全部提取DNA样本,基于隐孢子虫(Cryptosporidium)SSU rDNA序列进行PCR检测,序列比对分析后进行隐孢子虫种类鉴定;采用χ^(2)检验法比较不同规模化养殖场双峰驼隐孢子虫的感染率差异;将被鉴定为微小隐孢子虫(Cryptosporidium parvum,C.parvum)阳性的DNA样本,基于gp60基因序列进行PCR检测,序列比对分析后进行微小隐孢子虫亚型鉴定。[结果]516份双峰驼粪便DNA样本中检测出34份隐孢子虫阳性,总体感染率为6.59%(34/516),以阿恰勒镇养殖场2的双峰驼隐孢子虫感染率最高,为9.57%(9/94);不同养殖场双峰驼隐孢子虫的感染率统计学差异显著(χ^(2)=5.497,P<0.05)。基于SSU rDNA序列,34条隐孢子虫序列经比对分析,鉴定出3种隐孢子虫,分别为安氏隐孢子虫(n=29)、隐孢子虫大鼠基因型Ⅳ(n=1)和微小隐孢子虫(n=4);基于gp60基因序列,4条微小隐孢子虫序列经比对分析,均鉴定为Ⅰd-like-A21亚型,具有独特的遗传进化特点。[结论]柯坪县双峰驼隐孢子虫感染率较低,其感染的微小隐孢子虫具有独特亚型。研究结果为我国双峰驼源隐孢子虫种类分布特征和遗传进化提供了基础数据。

公驼发情期低摄食却仍能维持旺盛体力的潜在机制研究

【目的】利用蛋白组学的方法研究双峰公驼在发情期体重迅速下降但体力仍很旺盛的缘由。【方法】采集公驼发情期和非发情期血液样本为对照,对两个时期的样本进行蛋白表达定量分析,以揭示发情期与非发情期之间蛋白表达的差异。【结果】结果显示,在发情期和非发情期血液样本中,有26种蛋白表达量存在差异,其中在发情期4种蛋白表达量升高,22种蛋白表达量降低;COG分析发现,差异表达蛋白主要涉及细胞分裂、细胞周期、蛋白质周转,翻译后修饰等生物过程;GO分析揭示了蛋白质多种功能的存在,如生物糖原合成、免疫应答、细胞粘着、蛋白聚合、血板活化、信号传递、基微管合成、血液凝固等;KEGG分析发现存在与组织重塑有关的信号通路,以及与重要生物学过程有关的细胞基质黏附通路,如细胞运动、细胞增殖、细胞分化、基因表达调控和细胞存活等,同时还发现了磷脂酰肌醇3′-激酶(PI3K)-Akt信号通路。【结论】研究结果表明,GYS、PPP1R3G、AGP、mTOR、PI3k等蛋白质与公驼发情期低摄食仍能保持充沛精力密切相关。

青海骆驼线粒体DNA D-loop区遗传多样性研究

旨在研究青海骆驼遗传多样性,揭示青海骆驼的母系起源进化。采集柴达木地区3个青海骆驼类群耳组织样品88份并提取基因组DNA,利用PCR扩增和基因测序分析线粒体DNA D-loop序列,并与蒙古、内蒙古双峰驼进行比较。结果表明:D-loop序列中共检测到96个多态位点,定义了27种单倍型。3个青海骆驼类群占有16个单倍型,而蒙古双峰驼独有7个单倍型,内蒙古双峰驼独有4个单倍型。系统发育分析显示出两个独立的分支:第一支为青海骆驼3个类群与蒙古双峰驼,且德令哈双峰驼与其他两个类群间存在明显界限;第二支为内蒙古双峰驼。青海骆驼与内蒙古双峰驼的遗传距离均较远,但与蒙古双峰驼的遗传距离较近。因此,青海骆驼母系起源更倾向于蒙古双峰驼而非内蒙古双峰驼。

新疆地区塔里木双峰驼泌乳性能分析

【目的】旨在探究塔里木双峰驼的泌乳性能,为其开发与利用提供数据支持。【方法】本研究收集了2023年6—10月柯坪县畜牧科技发展有限公司(以下简称为畜牧公司)和新疆优驼骆驼养殖业农民专业合作社(以下简称为优驼公司)的823条产奶数据及10月份乳成分数据,包括乳蛋白率、乳脂率、乳糖率、总固体、乳尿素氮、体细胞数等。运用描述性统计分析、独立样本t检验和相关性分析等方法开展研究。【结果】结果显示,8月日均产奶量最高,为3.00kg,10月日均产奶量最低,为2.47 kg。乳脂率在4.04%~4.93%之间,乳糖率在5.17%~4.93%之间,乳蛋白率在4.06%~4.10%之间,总固体在14.14%~15.22%之间,体细胞数在27.28~34.24万个/mL之间,乳尿素氮的含量在44.34~49.24mg/dL之间。此外,乳脂率与乳蛋白率、总固体数呈极显著正相关(P<0.01),乳蛋白率与总固体数、乳尿素氮呈极显著正相关(P<0.01)。畜牧公司养殖的塔里木双峰驼的平均乳脂率、平均乳糖率、平均总固体数均极显著高于优驼公司(P<0.01),平均体细胞数高于优驼公司(P>0.05),平均乳尿素氮含量低于优驼公司(P>0.05)。【结论】本研究为塔里木双峰驼的饲养管理提供了理论依据。

乳用双峰驼产业化发展现状、思路与对策

骆驼产业是哈密市优势特色产业,经过近几年的养殖实践与培育发展,形成了企业、合作社从养殖端到加工端到市场的较为完善的产业链。文章通过对国家乳用双峰驼养殖标准化示范区乳用双峰驼产业化发展现状进行分析,梳理发展思路,提出针对性的发展对策,以期为骆驼产业发展提供理论支撑。

双峰驼乳功能蛋白及脂肪酸在泌乳期内的变化研究

为探究泌乳双峰骆驼乳功能性蛋白及脂肪酸变化规律,该研究动态追踪20峰泌乳双峰驼并采集泌乳期330 d内的驼乳,基于超高效液相色谱和气相色谱分析,检测α-乳白蛋白(α-lactalbumin,α-LA)、乳铁蛋白(lactoferrin,LF)、免疫球蛋白(immunoglobulin G,IgG)和溶菌酶4种目标功能蛋白及34种脂肪酸含量变化情况。结果表明,驼乳中共检测到4种目的功能蛋白及32种脂肪酸,色谱峰形尖锐、无杂峰干扰。驼乳4种功能蛋白含量在初乳期极显著高于常乳期(P<0.01),泌乳后期随着泌乳时间延长含量缓慢降低,功能蛋白间具有显著正相关性(P<0.05),其中α-乳白蛋白与IgG具有强正相关。脂肪酸含量在初乳期至常乳期30 d内波动明显,其余时期脂肪酸含量变化不大。长链脂肪酸、多不饱和脂肪酸和中链脂肪酸在泌乳期含量较为稳定;饱和脂肪酸和不饱和脂肪酸的比例合理,且饱和脂肪酸与不饱和脂肪酸及单不饱和脂肪酸变化趋势相反,在初乳期3~7 d内变化较为明显,在泌乳高峰期90~150 d波动极为明显,少数脂肪酸间具有较强的正相关。驼初乳功能蛋白与脂肪酸的组成及含量优于常乳,结果为深入研究双峰驼的泌乳生理特点及驼乳功能性制品的开发提供基础数据。

新疆阿勒泰市周边双峰驼饲养现状调查分析

随着阿勒泰地区骆驼产业的快速发展,为了解阿勒泰市周边双峰驼养殖现状,采用问卷调查和实地考察的方法对阿勒泰市周边双峰驼饲养现状进行调研。该研究从双峰驼的养殖规模、饲养方式、日粮组成、防疫措施以及死亡原因进行详实的统计分析,调查结果表明,在阿勒泰周边双峰驼饲养区,阿勒泰周边双峰驼的养殖以规模养殖和家庭养殖共存;养殖方式以放牧+补饲为主,占比43%;日粮组成以选择自配饲料为主;防疫措施以注射口蹄疫疫苗、使用驱虫药和饲养环境的消毒为主。该研究根据双峰驼饲养管理现状,提出其发展思路和对策,以期为阿勒泰地区特色产业的发展提供参考。

双峰驼乳营养成分测定与特征分析

双峰驼乳营养丰富,具有一定的保健功能和药用价值,但营养成分缺乏系统的检测和评价。从内蒙古呼伦贝尔市鄂温克旗、阿拉善左旗和乌海采集双峰驼乳155头份,同时作为相伴研究采集内蒙古荷斯坦牛乳101头份、萨能山羊乳34头份,采用国标法测定5种常规营养素、脂肪酸、氨基酸和矿物质,并进行描述性统计和主成分分析。结果表明,每个营养素组都可使3种家畜乳在三维空间以物种聚类,说明营养素都有物种特征,也提示营养素组可用于乳的物种真实性判别。双峰驼乳蛋白质、脂肪、乳糖、水分和灰分含量分别为(3.60±0.98)、(4.90±1.74)、(4.98±0.64)、(85.55±2.66)、(0.85±0.10)g/100g,其中脂肪、乳糖和灰分含量均显著高于萨能山羊乳和荷斯坦牛乳(P<0.05);双峰驼乳C15∶0、C16∶1、C18∶0、C18∶3n3显著高于萨能山羊乳和荷斯坦牛乳(P<0.05);双峰驼乳必需氨基酸Met、Ile和非必需氨基酸Asp、Pro、Tyr、Arg均显著高于荷斯坦牛乳和萨能山羊乳(P<0.05);Ca、Zn、I等矿物质含量均显著高于荷斯坦牛乳(P<0.05)。该研究获得了参考价值较高的双峰驼乳营养成分统计数据。