Bos taurus

Bos 金牛座
  • 文章类型: Journal Article
    Bostaurus以其对粗粒的耐受性而闻名,适应性,高温,湿度,湿度和抗病性。首先,牛被饲养为肉和奶,并确定与肉类生产相关性状相关的基因可以提高其整体生产力。这项研究的目的是确定基因组,分析进化,并探索金牛Pax基因家族的功能,为肉质牛育种提供新的分子靶标。在这项研究中,利用生物信息学技术从5个物种的基因组数据库中鉴定出44个Pax基因,表明牛科动物的亲缘关系相似。五只动物的Pax3和Pax7蛋白序列高度一致。总的来说,水牛的Pax基因对应于家畜。总之,水牛和家牛的Pax家族基因在Pax1/9,Pax2/5/8,Pax3/7和Pax4/6亚家族中的亲和力存在差异。我们认为Pax1/9对水牛和家畜的生长性状有影响。Pax3/7基因在水牛和家畜的进化中是保守的,可能是调节金牛芽孢杆菌生长的关键基因。Pax2/5/8亚族影响外套颜色,繁殖性能,和牛的产奶性能。Pax4/6亚家族对金牛座的乳脂百分比有影响。研究结果为理解进化论提供了理论依据,结构,金牛座Pax家族成员的功能特征以及分子遗传学和产肉金牛座物种的育种。
    Bos taurus is known for its tolerance of coarse grains, adaptability, high temperature, humidity, and disease resistance. Primarily, cattle are raised for their meat and milk, and pinpointing genes associated with traits relevant to meat production can enhance their overall productivity. The aim of this study was to identify the genome, analyze the evolution, and explore the function of the Pax gene family in B. taurus to provide a new molecular target for breeding in meat-quality-trait cattle. In this study, 44 Pax genes were identified from the genome database of five species using bioinformatics technology, indicating that the genetic relationships of bovids were similar. The Pax3 and Pax7 protein sequences of the five animals were highly consistent. In general, the Pax gene of the buffalo corresponds to the domestic cattle. In summary, there are differences in affinity between the Pax family genes of buffalo and domestic cattle in the Pax1/9, Pax2/5/8, Pax3/7, and Pax4/6 subfamilies. We believe that Pax1/9 has an effect on the growth traits of buffalo and domestic cattle. The Pax3/7 gene is conserved in the evolution of buffalo and domestic animals and may be a key gene regulating the growth of B. taurus. The Pax2/5/8 subfamily affects coat color, reproductive performance, and milk production performance in cattle. The Pax4/6 subfamily had an effect on the milk fat percentage of B. taurus. The results provide a theoretical basis for understanding the evolutionary, structural, and functional characteristics of the Pax family members of B. taurus and for molecular genetics and the breeding of meat-production B. taurus species.
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  • 文章类型: Journal Article
    背景:越来越多的证据表明输卵管细胞外囊泡(oEVs)在早期胚胎-母体对话中具有积极作用。然而,目前尚不清楚oEV是否有助于识别输卵管中胚胎的存在及其质量.因此,我们检查了牛输卵管上皮细胞(BOEC)分泌的oEV的分子货物是否根据良好的存在而有所不同(≥8个细胞,G)或较差(<8个细胞,P)优质胚胎。此外,分析了G和P胚胎之间RNA谱的差异,试图区分oEV和胚胎EV货物。
    方法:为此,在受精后53小时将原代BOEC与体外产生的胚胎(IVP)共培养,如下所示:BOEC与G胚胎(BGE);BOEC与P胚胎(BPE);单独的G胚胎(GE);单独的P胚胎(PE);单独的BOEC(B)和培养基对照(M)。共培养24小时后,收集所有组的条件培养基,并分离和表征EV.通过小RNA测序进行EV和胚胎的MicroRNA分析。
    结果:在电动汽车中,鉴定了84个miRNA,具有8个差异丰富的(DA)miRNA的BGE与B和4用于BPE与B(P值<0.01)。在胚胎中,鉴定了187个miRNA,有12个DAmiRNA用于BGE与BPE,3为Gvs.P,8为BGEvs.GE,和11为BPEvs.PE(P值<0.01)。
    结论:这些结果表明,oEV参与输卵管-胚胎识别,并指出在早期胚胎发育过程中具有信号传导和支持作用的特定miRNA。
    BACKGROUND: Increasing evidence points to an active role of oviductal extracellular vesicles (oEVs) in the early embryo-maternal dialogue. However, it remains unclear whether oEVs contribute to the recognition of the presence of embryos and their quality in the oviduct. Hence, we examined whether the molecular cargo of oEVs secreted by bovine oviduct epithelial cells (BOEC) differs depending on the presence of good (≥ 8 cells, G) or poor (< 8 cells, P) quality embryos. In addition, differences in RNA profiles between G and P embryos were analyzed in attempt to distinguish oEVs and embryonic EVs cargos.
    METHODS: For this purpose, primary BOEC were co-cultured with in vitro produced embryos (IVP) 53 h post fertilization as follows: BOEC with G embryos (BGE); BOEC with P embryos (BPE); G embryos alone (GE); P embryos alone (PE); BOEC alone (B) and medium control (M). After 24 h of co-culture, conditioned media were collected from all groups and EVs were isolated and characterized. MicroRNA profiling of EVs and embryos was performed by small RNA-sequencing.
    RESULTS: In EVs, 84 miRNAs were identified, with 8 differentially abundant (DA) miRNAs for BGE vs. B and 4 for BPE vs. B (P-value < 0.01). In embryos, 187 miRNAs were identified, with 12 DA miRNAs for BGE vs. BPE, 3 for G vs. P, 8 for BGE vs. GE, and 11 for BPE vs. PE (P-value < 0.01).
    CONCLUSIONS: These results indicated that oEVs are involved in the oviductal-embryo recognition and pointed to specific miRNAs with signaling and supporting roles during early embryo development.
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  • 文章类型: Journal Article
    大量研究表明,LCORL-NCAPG基因座的遗传变异与肉牛的生长性状密切相关。然而,一个致病的分子变异尚未被鉴定。要定义所有可能的候选变体,对34只夏洛来的小牛进行了全基因组测序,包括17个与增长相关的长程单倍型纯合(QQ)和17个该区域潜在祖先单倍型纯合(qq)。Q单倍型被细化到chr6:37,199,897-38,014,080之间的814kb区域,并包含在qq个体中未发现的218种变体。这些变体包括NCAPG内含子中的插入,先前记录的NCAPG突变(rs109570900),LCORL中的两个编码序列突变(rs109696064和rs384548488),和15个位于ATAC峰内的变体,其被预测影响转录因子结合。值得注意的是,rs384548488是移码变体,可能导致LCORL的长同种型的功能丧失。为了测试LCORL的编码序列变体与表型的关联,对来自五个种群的405头牛进行了基因分型。这两个变体处于完全连锁不平衡状态。对包含QQ动物的三个种群的统计学分析显示,基因型和出生体重具有显着(p<0.05)关联。活重,屠体重量,臀部高度,和平均每日收益。这些发现肯定了该基因座与肉牛生长之间的联系,并描述了定义单倍型的DNA变体。然而,需要进一步的研究来定义真正的致病突变.
    Numerous studies have shown genetic variation at the LCORL-NCAPG locus is strongly associated with growth traits in beef cattle. However, a causative molecular variant has yet to be identified. To define all possible candidate variants, 34 Charolais-sired calves were whole-genome sequenced, including 17 homozygous for a long-range haplotype associated with increased growth (QQ) and 17 homozygous for potential ancestral haplotypes for this region (qq). The Q haplotype was refined to an 814 kb region between chr6:37,199,897-38,014,080 and contained 218 variants not found in qq individuals. These variants include an insertion in an intron of NCAPG, a previously documented mutation in NCAPG (rs109570900), two coding sequence mutations in LCORL (rs109696064 and rs384548488), and 15 variants located within ATAC peaks that were predicted to affect transcription factor binding. Notably, rs384548488 is a frameshift variant likely resulting in loss of function for long isoforms of LCORL. To test the association of the coding sequence variants of LCORL with phenotype, 405 cattle from five populations were genotyped. The two variants were in complete linkage disequilibrium. Statistical analysis of the three populations that contained QQ animals revealed significant (p < 0.05) associations with genotype and birth weight, live weight, carcass weight, hip height, and average daily gain. These findings affirm the link between this locus and growth in beef cattle and describe DNA variants that define the haplotype. However, further studies will be required to define the true causative mutation.
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  • 文章类型: Journal Article
    单核苷酸多态性(SNP)在畜牧业和生产中的潜在应用是动物品种的鉴定。在这项研究中,使用选定的标记选择方法和使用Illumina牛SNP50BeadChip为属于十个牛品种的个体获得的基因型数据,开发了包含信息最丰富的SNP标记的减少的小组。通过在GeneClass2中实施的三种分配算法检查了所选SNP面板对所研究个体有效和可靠地分配到起源品种的适用性。所研究的品种集包括遗传资源保护计划下的波兰本地品种和全球范围内的高产品种。对于所有测试的标记选择方法(“delta”和两个基于FST的变体),使用了两种单独的标记分类方法学方法,并分别使用96、192和288个SNP创建了三个标记组,以确定所研究品种的有效分化所需的标记的最小数量。此外,研究了最有效的标记组对评估所分析品种的种群结构和遗传多样性的有用性。进行的分析表明,可以使用中密度基因型微阵列中的SNP子集来区分波兰饲养的牛的品种并分析其遗传结构。
    A potential application of single nucleotide polymorphisms (SNPs) in animal husbandry and production is identification of the animal breed. In this study, using chosen marker selection methods and genotypic data obtained with the use of Illumina Bovine SNP50 BeadChip for individuals belonging to ten cattle breeds, the reduced panels containing the most informative SNP markers were developed. The suitability of selected SNP panels for the effective and reliable assignment of the studied individuals to the breed of origin was checked by three allocation algorithms implemented in GeneClass 2. The studied breeds set included both Polish-native breeds under the genetic resources conservation programs and highly productive breeds with a global range. For all of the tested marker selection methods (\"delta\" and two FST-based variants), two separate methodological approaches of marker assortment were used and three marker panels were created with 96, 192, and 288 SNPs respectively, to determine the minimum number of markers required for effective differentiation of the studied breeds. Moreover, the usefulness of the most effective panels of markers to assess the population structure and genetic diversity of the analyzed breeds was examined. The conducted analyses showed the possibility of using SNP subsets from medium-density genotypic microarrays to distinguish breeds of cattle kept in Poland and to analyze their genetic structure.
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  • 文章类型: Journal Article
    据报道,有13头美国赫里福德牛在约12个月大时失明,推测发病。所有盲牛都有一个共同的祖先,通过母系和父系,暗示了隐性遗传起源.鉴于谱系关系和新的表型,我们对与失明相关的眼病理变化进行了表征,并鉴定了相关基因变异.5头盲牛的眼科检查显示视网膜变性。组织学上,2头盲牛的视网膜感光层丧失。对7头盲牛和9头未受影响的亲戚进行的全基因组测序(WGS)显示,在类脂褐变神经元3(CLN3;chr25g.26043843del)中存在1bp的移码缺失,其中盲牛是纯合的,其父母是杂合的。预测17的外显子16中鉴定的变体截短编码的蛋白质(p。Pro369Argfs*8)battenin,参与感光层维持所需的溶酶体功能。在462头牛中,只有盲牛是纯合的缺失。对>5,800只动物的WGS数据的查询进一步揭示了该变体仅在相关的赫里福德牛中观察到。CLN3的突变与人类幼年神经元类脂褐菌病(JNCL)有关,或巴顿病,导致早发性视网膜变性和病变与我们病例中观察到的相似。我们的数据支持CLN3的移码变体是这些赫里福德牛失明的原因,并提供了该基因在视网膜病变中的作用的额外证据,可能作为人类非综合征JNCL的模型。
    Thirteen American Hereford cattle were reported blind with presumed onset when ~12-mo-old. All blind cattle shared a common ancestor through both the maternal and paternal pedigrees, suggesting a recessive genetic origin. Given the pedigree relationships and novel phenotype, we characterized the ophthalmo-pathologic changes associated with blindness and identified the responsible gene variant. Ophthalmologic examinations of 5 blind cattle revealed retinal degeneration. Histologically, 2 blind cattle had loss of the retinal photoreceptor layer. Whole-genome sequencing (WGS) of 7 blind cattle and 9 unaffected relatives revealed a 1-bp frameshift deletion in ceroid lipofuscinosis neuronal 3 (CLN3; chr25 g.26043843del) for which the blind cattle were homozygous and their parents heterozygous. The identified variant in exon 16 of 17 is predicted to truncate the encoded protein (p. Pro369Argfs*8) battenin, which is involved in lysosomal function necessary for photoreceptor layer maintenance. Of 462 cattle genotyped, only blind cattle were homozygous for the deletion. A query of WGS data of > 5,800 animals further revealed that the variant was only observed in related Hereford cattle. Mutations in CLN3 are associated with human juvenile neuronal ceroid lipofuscinosis (JNCL), or Batten disease, which results in early-onset retinal degeneration and lesions similar to those observed in our cases. Our data support the frameshift variant of CLN3 as causative of blindness in these Hereford cattle, and provide additional evidence of the role of this gene in retinal lesions, possibly as a model for human non-syndromic JNCL.
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  • 文章类型: Journal Article
    环状RNA(circularRNAs)是独特的非编码RNA分子,值得注意的是它们的共价闭环结构,在调节各种生物过程中的基因表达中起着至关重要的作用。这篇综述综合了三个关键牲畜物种中circRNAs的现有知识:Bostaurus(牛),Ovisaries(绵羊),和Caprahircus(山羊)。它侧重于它们的功能重要性和作为疾病检测生物标志物的新兴潜力,应激反应,和整体生理健康。具体来说,它深入研究了这些物种中circRNAs的表达和功能,特别注意对牲畜生产力至关重要的特征,如牛奶生产,肉质,肌肉发育,羊毛生产,免疫反应,等。我们还解决了当前在circRNA研究中面临的挑战,包括需要标准化的方法和更广泛的研究。通过提供对circRNAs调控的分子机制的见解,这篇评论强调了它们在畜牧业中的科学和经济意义。circRNAs改善动物健康管理和动物源性产品质量的潜力与消费者对动物福利和可持续性的日益关注相一致。因此,本文旨在指导未来的研究方向,同时支持畜牧业管理和育种创新策略的发展。
    Circular RNAs (circRNAs) are unique noncoding RNA molecules, notable for their covalent closed-loop structures, which play a crucial role in regulating gene expression across a variety of biological processes. This review comprehensively synthesizes the existing knowledge of circRNAs in three key livestock species: Bos taurus (cattle), Ovis aries (sheep), and Capra hircus (goats). It focuses on their functional importance and emerging potential as biomarkers for disease detection, stress response, and overall physiological health. Specifically, it delves into the expression and functionality of circRNAs in these species, paying special attention to traits critical to livestock productivity such as milk production, meat quality, muscle development, wool production, immune responses, etc. We also address the current challenges faced in circRNA research, including the need for standardized methodologies and broader studies. By providing insights into the molecular mechanisms regulated by circRNAs, this review underscores their scientific and economic relevance in the livestock industry. The potential of circRNAs to improve animal health management and the quality of animal-derived products aligns with growing consumer concerns for animal welfare and sustainability. Thus, this paper aims to guide future research directions while supporting the development of innovative strategies in livestock management and breeding.
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  • 文章类型: Journal Article
    储藏稻谷的主要害虫是米稻米(水稻象鼻虫)。农民每年生产大量的水稻,但是由于储存设施不足和处理不当带来的虫害袭击,大部分被浪费了。在这项研究中,在Susscrofa(猪)的股骨中评估了生物炭对米曲霉的杀虫效果,鸡(鸡),和Bostaurus(牛)。成人死亡率,成人的出现,减肥,种子损伤,和象鼻虫穿孔指数是评价指标之一。为了应用生物炭,使用不同的剂量:0.2、0.4、0.6、0.8和1.0g/20g水稻。在这项工作中,通过测试所有动物骨骼生物炭的杀虫活性,水稻上成年米曲霉的种群显着减少(p<0.05)。随着剂量的增加,动物骨骼生物炭变得更加有效。治疗24小时后,猪生物炭诱导的成年米曲霉死亡率为36.67%,其次是奶牛生物炭的水稻象鼻虫死亡率为20%。牛的致命剂量,猪,处理24小时后,成年米曲霉种群的50%(LD50)的鸡生物炭为0.83g,0.43g,和0.90g,分别。应该鼓励利用由动物骨骼制成的生物炭来对抗储存稻谷中的米曲霉。它们可以用作绿色控制措施,以减少在环境中使用合成化学杀虫剂所带来的风险。
    The main insect pest of stored paddy is Sitophilus oryzae (rice weevil). Huge amounts of rice are produced by peasant farmers each year, but the bulk of it is wasted due to insufficient storage facilities and insect pest attacks brought on by careless handling. In this study, the insecticidal effects of biochar on S. oryzae were assessed in femurs from Sus scrofa (pig), Gallus gallus (chicken), and Bos taurus (cattle). Adult mortality, adult emergence, weight loss, seed damage, and weevil perforation index are among the indicators evaluated. To apply the biochar, different dosages were used: 0.2, 0.4, 0.6, 0.8, and 1.0 g/20 g of paddy rice. In this work, the population of adult S. oryzae on paddy rice was dramatically reduced by all animal bones biochar tested for insecticidal activity (p < 0.05). The animal bone biochar became more effective with increasing dosage. After 24 h of treatment, pig biochar induced a 36.67 % mortality rate of adult S. oryzae, followed by cow biochar\'s 20 % mortality rate of rice weevils. The fatal dose of cattle, pig, and chicken biochar at which 50 % (LD50) of the population of adult S. oryzae responded after 24 h of treatment were 0.83g, 0.43g, and 0.90g, respectively. It should be encouraged to utilize biochar made from animal bones to combat the S. oryzae in stored paddy. They could be utilized as a green control measure to lessen the risk brought on by the usage of synthetic chemical insecticides in the environment.
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  • 文章类型: Journal Article
    热带嗜血原虫病是巴基斯坦最普遍的嗜血原虫病。
    该研究旨在调查旁遮普邦不同农业气候地区的牛中环流Theileria的流行病学和进化关系,巴基斯坦。
    使用多阶段采样方法从Sargodha(n=400)和Multan(n=400)地区的无症状牛(n=480)和水牛(n=320)中收集了800份血液样本。评估样品的血液涂片显微镜和基于细胞色素b基因的PCR。从每个地区的每个工会理事会收集了20个样本。
    通过血液涂片分析和PCR确定,牛的环状衣原体感染的总体患病率分别为9%和17.13%,分别。通过血涂片筛查,牛和水牛的疾病阳性率分别为10.21%和20.42%和7.19%,12.19%通过PCR。Sargodha和Multan地区基于PCR的总体患病率为19%和15.25%,分别。没有农村家禽,蜱虫侵扰,有蜱传疾病史对牛有显著影响。tick虫感染和年龄是水牛的主要统计学上显着疾病决定因素。细胞色素b基因的进化分析表明,感染水牛的巴基斯坦分离株与伊朗的分离株有关,印度,埃及,苏丹。牛的分离株在基因上与巴基斯坦的分离株接近,印度,伊朗,伊拉克,和土耳其。
    可以得出结论,生物和非生物因素有助于疾病的发生。目前的研究将有助于制定控制策略,以防止重大的经济损失。
    UNASSIGNED: Tropical theileriosis is the most prevalent hemoprotozoan disease in Pakistan.
    UNASSIGNED: The study aimed to investigate the epidemiology and evolutionary relationship of Theileria annulata in bovines in diverse agro-climatic regions of Punjab, Pakistan.
    UNASSIGNED: 800 blood specimens were collected from asymptomatic cattle (n=480) and buffaloes (n=320) using a multistage sampling method from Sargodha (n=400) and Multan (n=400) districts. The samples were assessed for blood smear microscopy and cytochrome b gene based PCR. Twenty samples were collected from each union council of each district.
    UNASSIGNED: The overall prevalence of T. annulata infection in bovines was 9% and 17.13% as determined by blood smear analysis and PCR, respectively. The disease positivity in cattle and buffaloes was respectively 10.21% and 20.42% by blood smear screening and 7.19%, 12.19% by PCR. The overall PCR based prevalence in the Sargodha and Multan districts was 19% and 15.25%, respectively. Absence of rural poultry, tick infestation, and a history of tick-borne diseases had significant effect in cattle. Tick infestation and age were the main statistically significant disease determinants in buffaloes. The evolutionary analysis of the cytochrome b gene showed that the Pakistani isolate infecting buffalo was related to those from Iran, India, Egypt, and Sudan. The isolate from cattle was genetically close to those from Pakistan, India, Iran, Iraq, and Turkey.
    UNASSIGNED: It can be concluded that biotic and abiotic factors contribute to disease occurrence. The current study will help to devise control strategies to prevent substantial economic losses.
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  • 文章类型: Journal Article
    本研究旨在评估摄入量,在过渡时期,荷斯坦和½Holstein½Gyr(Girolando-F1)奶牛的能量和氮平衡以及甲烷排放。24头母牛(12头荷斯坦和12头Girolando-F1)用于评估采食量,表观消化率,产热和甲烷排放,分两个时期进行:28至19天产卵前和15至23天产卵后。使用完全随机的设计,并考虑到遗传群体的主要影响,通过ANOVA在时期内(产卵前和产卵后)分析数据。与Holstein相比,Girolando-F1奶牛的身体状况评分(BCS)更高。在产卵前,遗传群体之间没有差异,除了荷斯坦奶牛每公斤代谢体重的最高产热。产牛后,荷斯坦奶牛有更多的DM摄入量,氮,每千克BW的NDF和每千克代谢体重产生更多的热量。与Girolando-F1奶牛相比,荷斯坦奶牛的牛奶和脂肪校正牛奶(FCM4%)更多。荷斯坦奶牛每单位体重和代谢重量的甲烷排放量较高。与Girolando-F1母牛相比,荷斯坦每公斤牛奶和每公斤FCM4%的肠溶甲烷排放量往往较低。在产卵前后,荷斯坦和Girolando-F1的氮和能量保持力相似。尽管BCS存在差异,QI,和牛奶产量,Girolando-F1和Holstein奶牛的整体能源效率相似,尽管荷斯坦奶牛每公斤合格产品(牛奶)的甲烷排放量往往较少。
    This study aimed to evaluate intake, energy and nitrogen balance as well as methane emission in Holstein and ½ Holstein ½ Gyr (Girolando-F1) cows during the transition period. Twenty-four cows (12 Holstein and 12 Girolando-F1) were used to evaluate feed intake, apparent digestibility, heat production and methane emission, carried out in two periods: from 28 to 19 days pre-calving and from 15 to 23 days post-calving. A completely randomised design was used and data were analysed by ANOVA within periods (pre- and post-calving) considering the main effect of genetic groups. Girolando-F1cows presented greater body condition score (BCS) compared with Holstein. During pre-calving, there were no differences between genetic groups, except for highest heat production per kilogram of metabolic body weight for Holstein cows. After calving, Holstein cows had greater intake of DM, nitrogen, NDF per kg of BW and produced more heat per kg of metabolic body weight. Holstein cows yielded more milk and fat-corrected milk (FCM4%) compared with Girolando-F1 cows. Holstein cows presented higher methane emission per unit of BW and of metabolic weight. Emissions of enteric methane per kilogram of milk and per kilogram of FCM4% tended to be lower for Holstein compared with Girolando-F1 cows. Nitrogen and energy retention were similar for both Holstein and Girolando-F1 at pre- and post-calving. Despite differences in BCS, DMI, and milk yield, Girolando-F1 and Holstein cows present overall similar energy efficiency, albeit Holstein cows tended to present less methane emission per kg of eligible product (milk).
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  • 文章类型: Journal Article
    在美国,产志贺毒素的大肠杆菌(STEC)和单核细胞增生李斯特菌通常是导致严重食源性疾病的原因。美国食品安全检验局使用的当前鉴定方法需要至少四天来鉴定STEC,而单核细胞增生李斯特菌则需要六天。采用长读,用于食品安全检测的全基因组测序可以显着减少鉴定所需的时间,但是方法开发成本很高。因此,该项目的目标是使用NanoSim-H软件模拟牛津纳米孔测序读数,以评估基于测序的食源性病原体检测的可行性并指导实验设计.模拟STEC的测序读数,L.单核细胞增生,以及使用NanoSim-H的STEC和Bos金牛座基因组的1:1组合。至少需要2500个模拟读数来鉴定STEC中靶向的七个感兴趣的基因,至少需要500个读数来检测单核细胞增生李斯特菌靶向的基因。估计30x的基因组覆盖率为21,521,STEC和单核细胞增生李斯特菌的读数为11,802,分别。由于引入错误,从两种细菌模拟的大约5-6%的读段未与它们各自的参考基因组对齐。对于STEC和B.taurus1:1基因组混合物,所有感兴趣的基因都被检测到1,000,000个读数,但获得的覆盖率不到1x。结果表明,通过长读数测序检测食源性病原体需要样品富集,但这仍然会减少当前方法所需的时间。此外,模拟数据将有助于减少与实验室实验相关的时间和费用。
    Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes are routinely responsible for severe foodborne illnesses in the United States. Current identification methods utilized by the U.S. Food Safety Inspection Service require at least four days to identify STEC and six days for L. monocytogenes. Adoption of long-read, whole genome sequencing for food safety testing could significantly reduce the time needed for identification, but method development costs are high. Therefore, the goal of this project was to use NanoSim-H software to simulate Oxford Nanopore sequencing reads to assess the feasibility of sequencing-based foodborne pathogen detection and guide experimental design. Sequencing reads were simulated for STEC, L. monocytogenes, and a 1:1 combination of STEC and Bos taurus genomes using NanoSim-H. At least 2500 simulated reads were needed to identify the seven genes of interest targeted in STEC, and at least 500 reads were needed to detect the gene targeted in L. monocytogenes. Genome coverage of 30x was estimated at 21,521, and 11,802 reads for STEC and L. monocytogenes, respectively. Approximately 5-6% of reads simulated from both bacteria did not align with their respective reference genomes due to the introduction of errors. For the STEC and B. taurus 1:1 genome mixture, all genes of interest were detected with 1,000,000 reads, but less than 1x coverage was obtained. The results suggested sample enrichment would be necessary to detect foodborne pathogens with long-read sequencing, but this would still decrease the time needed from current methods. Additionally, simulation data will be useful for reducing the time and expense associated with laboratory experimentation.
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