Improvement on molecular sex identification primers for Passeriform bird species

The primer pair sex1/sex2,which can be widely applied for sex identification in Galliform species,was used to determine the sex of 17 Passeriform species.As CHD-W fragments tended to be preferentially amplified,which may cause unnecessary misidentification in bird species with little difference between CHD-Z and CHD-W,we modified sex1 and sex2,obtaining sex1' and sex-mix respectively.Primer sets were then recombined to conduct sex identification.After testing several Passeriforme birds of known sex,we found that the primer pair sex1'/sex2 was better at limiting the preferential amplification of CHD-W fragments.As they are being frequently used in sex allocation study of Aegithalos concinnus and song learning research of Lonchura striata,we can expect more applications of this primer pair to further studies in Passeriformes.

Individual Identification of Dairy Cows Based on Deep Feature Extrac-tion and Matching

Individual identification of dairy cows is the prerequisite for automatic analysis and intelligent perception of dairy cows'behavior.At present,individual identification of dairy cows based on deep convolutional neural network had the disadvantages in prolonged training at the additions of new cows samples.Therefore,a cow individual identification framework was proposed based on deep feature extraction and matching,and the individual identification of dairy cows based on this framework could avoid repeated training.Firstly,the trained convolutional neural network model was used as the feature extractor;secondly,the feature extraction was used to extract features and stored the features into the template feature library to complete the enrollment;finally,the identifies of dairy cows were identified.Based on this framework,when new cows joined the herd,enrollment could be completed quickly.In order to evaluate the application performance of this method in closed-set and open-set individual identification of dairy cows,back images of 524 cows were collected,among which the back images of 150 cows were selected as the training data to train feature extractor.The data of the remaining 374 cows were used to generate the template data set and the data to be identified.The experiment results showed that in the closed-set individual identification of dairy cows,the highest identification accuracy of top-1 was 99.73%,the highest identification accuracy from top-2 to top-5 was 100%,and the identification time of a single cow was 0.601 s,this method was verified to be effective.In the open-set individual identification of dairy cows,the recall was 90.38%,and the accuracy was 89.46%.When false accept rate(FAR)=0.05,true accept rate(TAR)=84.07%,this method was verified that the application had certain research value in open-set individual identification of dairy cows,which provided a certain idea for the application of individual identification in the field of intelligent animal husbandry.

Morphological traits for sex identification of the oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)

The oriental armyworm,Mythimna separata(Lepidoptera:Noctuidae)is a harmful pest which causes heavy loss in agricultural crops.Sex identification of M.separata is very important for understanding the biology and management of this pest.Development of fast and convenient methods to identify M.separata female and male is the necessary prerequisite for con ducting gender-related studies.In this study,new methods to differentiate sex of M.separata at pupal and adult stages were investigated.Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male.Genital openings are important characters for the female and male pupae differentiation.For female pupa,it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture;for male pupa,it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps.Bristle numbers are another indicator for sex differentiation of adults.Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body.We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.

Progress in Sex Identification of Early Embryo of Livestock

Principle, research, and problems of techniques for sex identification of early embryo of livestock were discussed in detail from aspects of main ways for sex control, main methods for sex identification of early embryo, and application of sex identification in livestock production. And development prospects were expected.

Morphometric and Volumetric Evaluation of Maxillary Sinus as an Aid in Sex Identification in Forensic Odontology-A Cone-Beam Computed Tomography Study

Background:Various methods are used in forensic odontology for the purpose of sex and personal identification.Some of them include cheiloscopy,rugoscopy,mandibular measurements,and study of paranasal sinuses.In our study,we have used maxillary sinus as an aid in sex identification.For this purpose,we have evaluated the morphometric and volumetric measurements of the maxillary sinus using a 3-Dimensional imaging modality,Cone Beam Computed Tomography.Aims and Objectives:Sex and age identification are important in the process of identification of both the living and the dead.Hence,this transverse retrospective study was done to appraise the precision of the dimensions and volume of the maxillary sinus to aid in sex and age identification.Material and Methods:Eighty full Field of View(FOV)Cone-Beam Computed Tomography(CBCT)images were retrieved from the archives which were matched with age and sex.The maxillary sinuses on either side were measured mediolaterally in the axial section and supero-inferiorly in the coronal section.The volumetric analysis of the sinus was done in the sagittal section.The sex was classified using discriminant function analysis and the differences were compared using independent t-test.The differences with age were analyzed using one-way ANOVA.Results:Independent t-test was done for comparison of the sinuses between both sexes.Maxillary sinuses measurements were larger among the males both superior-inferiorly on either side(P<0.001)and mediolaterally on the right(P<0.049)showed statistically significant values.A significant correlation existed while comparing the maxillary sinus volume among both the sexes on either side(P<0.001 and<0.002 in the case of right and left respectively).On comparing the supero-inferior and mediolateral measurements and the maxillary sinus volume on either side among the various age groups,the values were statistically insignificant.Present study shows that the maxillary sinus measurements using Cone-beam computed tomography were diagnostic in the identification of sex but not in age estimation and can be used in forensic dentistry.

Sex-related gene and sex identification of Crested Ibis Nipponia nippon (Ciconiifor-mes: Threskiornithidae)

The Crested Ibis (Nipponia nippon) is a critical endangered species of the world. At present, a reintroduction program will be conducted to save this species essentially. However, because the Crested Ibise is a sexual alike bird, it is very difficult to identify the sex from the morphological character. In order to identify the sex easily and select the right individuals for captive breeding and reintroduction, the sex related gene on W chromosome was amplified and the sex of three Crested Ibises were also identified in the present study. The 262bp fragment was also sequenced, and we found that there were 13 different nucleotide sites with 2.25 of transition/transversion based on the comparison with that of the Oriental White Stork. The sequence will also provide a theoretic base for further designing specific primer for sex related gene in the Crested Ibis.

Mechanism of fluorescent cocoon sex identification for silkworms Bombyx mori

By using silkworms,Bombyx mori, fluorescent cocoon sex identification (FCSI) as an experimental material, direct fluorescence spectrometry of the cocoon surface indicates that the fluorescent color of silkworm cocoons is made up of two peaks of yellow and blue-purple fluorescence emission. The fluorescent difference between male and female cocoons is attributed to the differential absorption of yellow fluorescent substances by the midgut tissue of 5th instar female silkworms. Thin layer chromatography (TLC) and fluorescent spectra indicate that blue-purple fluorescent substances are composed of at least five blue-purple fluorescent pigments, and yellow fluorescent substances are made up of at least three. UV spectra and AlCl3 color reaction show that the three fluorescent yellow pigments are flavonoids or their glycosides. Silkworm FCSI is due to selective absorption or accumulation of the yellow fluorescent pigments by the posterior midgut cells of female 5th instar larvae. The cells of the FCSI silkworm midgut, especially the cylinder intestinal cells of the posterior midgut have a component which is a yellow fluorescent pigment-specific binding protein that may be vigorously expressed in the 5th instar larvae.

Identification of the Sex of Earlier Embryos from Generic Hybrids of Chicken-Quail by Wpkci

In this study, a protocol was developed to identify the sex of earlier embryos of chicken （♂）-quail （♀） hybrids and successfully tested the sex proportion of each period （66-120 h）. We acquired cross bred eggs by artificial insemination, hatched them in the same batch according to the standard hatching condition of chicken, and collected earlier living embryos at 66, 72, 78, 84, 90, 96, 102, 108, 114, and 120 h randomly. We adopted RT-PCR protocol and multiple PCR, made the known sex quail as the external control, employed fl-actin as the internal control, and used primers that were designed according to conservative area of gene Wpkci of quail to identify the sex of earlier hybrid embryos. The results indicated that the primer of Wpkci can be used to identify the sex of hybrid embryos accurately; there were more male than female in earlier embryos, the sex proportion of earlier embryos compared with academic numerical value was significantly different （P〈0.01）, and there was no difference between different periods （P〉0.05）. In the present study, we concluded that a simple, fast, credible and stable protocol to identify the sex of earlier hybrids embryos had been established by using primer of Wpkci; in earlier embryos, the death rate of female was higher than that of male and there was no fluctuant peak.

Sex Identification of Red-crowned Crane by the Polymerase Chain Reaction

Sex determining gene primers of Oriental White Stork were used to amplify sex-linked gene of the Red-crowned Crane′s W chromosome-specific by PCR for sex identification. The sexes of 7 couples of grown Red-crowned Cranes and 15 youngs were identified. Through DNA sequence analysis, the identity is 94.77% between Red-crowned Crane and Oriental White Stork. The results of this study suggest that the application of the polymerase chain reaction technique is practicable for determining sex in the Red-crowned Crane.

基于牛脸和躯干综合信息的奶牛个体识别研究

针对基于单一信息对奶牛个体身份识别精度低的问题,本文提出1种基于牛脸和躯干综合信息对奶牛个体身份识别的方法。在Mask R-CNN目标检测模型的基础上进行改进,将注意力机制模块引入到Mask R-CNN的ResNet50特征提取网络的输出阶段,能够在图像通道和空间上增强奶牛身份信息。针对奶牛不同部位,本文对改进前后的Mask R-CNN模型分别基于牛脸、基于躯干以及基于牛脸和躯干综合信息进行了相关实验。实验结果表明,原始Mask R-CNN模型基于牛脸和躯干综合信息进行奶牛个体识别,比单独基于牛脸或躯干的识别精度提高2.3%~3.7%。改进后的Mask R-CNN模型在自建奶牛图像数据集上的准确率达到了93.63%,mAP值达到92.16%,相较于原始Mask R-CNN,准确率提高了2.92%,mAP值提高了2.63%。本文方法能够实现对养殖场环境下奶牛个体身份的识别,可为奶牛的精准养殖提供技术支持。

基于3D CNN-BiLSTM-ATFA网络和步态特征的奶牛个体识别方法

针对基于花纹的奶牛个体识别中纯色或花纹较少的奶牛识别准确率较低的问题,本文提出一种基于步态特征的奶牛个体识别方法。首先,将DeepLabv3+语义分割算法的主干网络替换为MobileNetv2网络,并引入基于通道和空间的CBAM注意力机制,利用改进后模型分割出奶牛的剪影图。然后,将三维卷积神经网络(3D CNN)和双向长短期记忆网络(BiLSTM)构建为3D CNN-BiLSTM网络,并进一步集成自适应时间特征聚合模块(ATFA)生成3D CNN-BiLSTM-ATFA奶牛个体识别模型。最后,在30头奶牛的共1242条视频数据集上进行了奶牛个体识别实验。结果表明,改进后DeepLabv3+算法的平均像素准确率、平均交并比、准确率分别为99.02%、97.18%和99.71%。采用r3d_18作为3D CNN-BiLSTM-ATFA的主干网络效果最优。基于步态的奶牛个体识别平均准确率、灵敏度和精确度分别为94.58%、93.47%和95.94%。奶牛躯干和腿部不同部位进行加权特征融合的个体识别实验表明识别准确率还可进一步提高。奶牛跛足对步态识别效果影响较为明显,实验期间由健康变为跛足和一直跛足的奶牛个体识别准确率分别为89.39%和92.61%。本文研究结果可为奶牛的智能化个体识别提供技术参考。

铃斑翅夜蛾蛹及成虫性别的快速鉴定

本文通过观察比较铃斑翅夜蛾Serrodes campana Guenée蛹及成虫雌雄个体之间外部形态差异,总结出其蛹及成虫个体雌雄鉴别特征与区分方法,结果表明:雌蛹腹部末端第8腹节有生殖孔和产卵孔连接形成的纵裂缝,形成“人”字形,裂缝两侧平坦无突起;雄蛹第8腹节无裂缝,第9腹节靠近第10腹节中央有1纵裂缝,裂缝两侧各有1个半圆形瘤状突起。雌成虫前翅亚中褶内侧的黑斑外侧半圆形,前翅基线至外线间与前胸背板同色,腹部末端圆锥状;雄成虫前翅亚中褶内侧的黑斑外侧直线形,前翅基线至外线间浅黄色,颜色较前胸背板浅,腹部末端圆筒状。此方法简易有效,能极大提升野外区分蛹和成虫性别的效率,对研究其生殖生物学和后续提取性信息素意义重大。

湖南某规模奶牛场乳房炎病原菌的分离鉴定与耐药性分析

为明确湖南某规模奶牛场临床型乳房炎病原菌种类及耐药情况,对32份临床型乳房炎患牛乳样进行传统细菌培养、生化试验、16S rDNA核苷酸序列检测和药敏试验。结果共分离鉴定大肠杆菌23株,乳房链球菌12株,肺炎克雷伯氏菌5株,样品检出率分别为71.9%(23/32)、37.5%(12/32)、15.6%(5/32);药敏试验显示,大部分大肠杆菌和乳房链球菌对青霉素、阿莫西林、头孢他啶、庆大霉素、林可霉素耐药严重,耐药率达50%~86.9%,对亚胺培南、四环素及红霉素较敏感,敏感率达75%~82.6%;肺炎克雷伯氏菌对大部分抗生素耐药程度较低,对亚胺培南、阿米卡星、四环素、红霉素敏感率为100%。建议该养殖场抓好环境卫生、提高生物安全,建立科学、合理治疗方案。

39例性别重置术后法医临床性别鉴定案例分析

目的 分析易性症人群性别重置术后法医临床性别鉴定案例的特点,结合国内外性别变更政策及相关案例,探讨我国对性别重置术人群的性别鉴定要点,旨在规范相关鉴定。方法 收集复旦大学上海医学院司法鉴定中心近年39例法医临床性别鉴定案例,对性别比例、鉴定时间、手术地点、第二性征等进行统计分析。结果 跨性别女性案例占97.4%,手术地点在泰国者占94.9%,全部案例均在完成性别重置术后进行鉴定,均至少有一项第二性征与术后第一性征相符,辅助检查应用案例仅有2例。结论 法医临床性别鉴定应以第一性征、第二性征与变更后的性别相符为原则。

基于视频和BCE-YOLO模型的奶牛采食行为检测

【目的】动物采食行为是一个重要的动物福利评价指标,本研究旨在解决复杂饲养环境下奶牛采食行为识别精度差、特征提取不充分的问题,实现对奶牛采食行为的自动监控。【方法】本文提出了一种基于改进BCEYOLO模型的识别方法,该方法通过添加BiFormer、CoT、EMA 3个增强模块,提高YOLOv8模型特征提取的能力,进一步与优于Staple、SiameseRPN算法的Deep SORT算法结合,实现对奶牛采食时头部轨迹的跟踪。在奶牛采食过程的俯视和正视视频中提取11288张图像,按照6∶1比例分为训练集和测试集,构建采食数据集。【结果】改进的BCE-YOLO模型在前方和上方拍摄的数据集上精确度分别为77.73%、76.32%,召回率分别为82.57%、86.33%,平均精确度均值分别为83.70%、76.81%;相较于YOLOv8模型,整体性能提升6~8个百分点。Deep SORT算法相比于Staple、SiameseRPN算法,综合性能提高1~4个百分点;并且改进的BCE-YOLO模型与Deep SORT目标跟踪算法结合良好,能对奶牛采食行为进行准确跟踪且有效地抑制了奶牛ID(Identity document)的变更。【结论】本文提出的方法能有效解决复杂饲养环境下奶牛采食行为识别精度差、特征提取不充分的问题,为智能畜牧与精确养殖提供重要参考。

基于微卫星标记的江西桃红岭梅花鹿遗传多样性

梅花鹿(Cervus nippon)为国家一级重点保护野生动物,江西桃红岭梅花鹿华东亚种(华南梅花鹿,C.n.kopschi)是我国现存最大的梅花鹿种群。研究桃红岭梅花鹿遗传多样性,并阐明其遗传本底状况,对于提升华南梅花鹿遗传多样性水平和促进种群快速发展具有重要意义。本研究在桃红岭梅花鹿国家级自然保护区共采集疑似梅花鹿粪便样品108份。首先基于线粒体Cyt b基因扩增测序进行物种鉴定,并利用微卫星标记和SRY/ZFX性别标记对梅花鹿样品进行个体识别、遗传多样性分析和性别鉴定。结果表明,108份样品中有96份梅花鹿粪便样品,来源于63只不同个体,且雌雄性比为1.33∶1。7个微卫星位点的平均等位基因数、观测杂合度、期望杂合度和近交系数分别为3.714、0.712、0.602和−0.087,其中3个位点显著偏离Hardy−Weinberg平衡,均表现为杂合子过剩。本研究结果表明,桃红岭梅花鹿种群的遗传多样性处于中等水平,且存在一定程度的性比失衡,近交系数为负值表明其种群暂无近交风险。建议积极推进桃红岭梅花鹿人工繁育,加强对稀有等位基因的保护与管理,并适时采取个体迁移等手段促进桃红岭梅花鹿与其他梅花鹿种群的基因交流,以提升其种群遗传多样性水平和促进种群快速发展。

基于白尾海雕粪便样品的SSR引物及性别鉴定引物的开发

白尾海雕(Haliaeetus albicilla),是鹰形目鹰科海雕属的一种大型猛禽、国家一级保护动物,位于食物链顶端,对于维持生态平衡和生物多样性具有重要作用.每年冬季,大批白尾海雕在我国吉林省珲春市敬信湿地越冬,形成了国内最大的越冬种群.为了实现基于粪便样品开展白尾海雕种群遗传学等方面的研究的可行性,填补白尾海雕亚洲种群的研究空白,需要开发新的微卫星DNA,即SSR(simple sequence repeat)引物和性别鉴定引物.本研究以NCBI上公布的白尾海雕全基因组数据为基础,进行了SSR引物和性别鉴定引物的开发.随后,使用白尾海雕粪便样品对引物进行了筛选,最终获得9对SSR引物和2对性别鉴定引物.本研究为白尾海雕种群遗传学深入研究提供了丰富有效的SSR分子标记,也为白尾海雕性别分子鉴定提供了新的引物选择.

不同临床型奶牛生殖道乳酸菌的筛选鉴定及菌群多样性分析

本研究对新疆南疆地区不同临床型奶牛生殖道分泌物中的乳酸菌进行了筛选鉴定,并对其菌群进行了多样性分析,评估了其微生态体系。将南疆地区3头健康与3头患子宫内膜炎的奶牛分为健康组和患病组,分别采集其生殖道分泌物。通过分离纯化、镜检、16S rRNA测序、高通量测序对奶牛生殖道分泌物菌群进行分析。结果表明:革兰染色后镜检观察到球状形态的菌体,分离镜检初步得到17株革兰阳性菌。高通量测序共获得652 901条有效序列,质量过滤、去噪共获得5 372个扩增子序列变体(ASV)。在属分类学水平以棒状杆菌属差异性最为显著,乳酸菌层次以链球菌属差异性最为显著。Alpha多样性分析显示,健康组奶牛生殖道样本群落丰富度和多样性均远高于患病组,且差异显著(P=0.05)。Beta多样性分析结果显示,层次聚类后6个样本分别按组聚集为2簇,表明2组样本组内差异较小,组间菌落相似性差异较大,说明不同临床型奶牛生殖道分泌物菌群在属分类学水平存在差异,健康型奶牛生殖道菌群丰富度和多样性均远高于患病组,其差异与子宫内膜炎的相关性有待进一步研究。

米缟螟蛹及成虫雌雄个体的快速鉴别方法

为建立一种快速鉴别米缟螟Aglossa dimidiata蛹及成虫雌雄的方法,分别利用超景深数码显微系统及扫描电镜对米缟螟蛹和成虫的外部特征进行观察并统计。结果表明:雌蛹第8腹节腹面上有一扁平的纵裂缝为生殖孔,生殖孔离肛门的距离较远,分节不明显;而雄蛹的生殖孔在第9腹节,生殖孔两侧各有一个半瘤状突起,且生殖孔离肛门的距离较近,分节较明显。雌性成虫腹部较为粗壮,腹部末端呈圆筒状且有圆形环毛,雄性成虫腹部至末端较为瘦小,呈倒三角状且有味刷。利用该方法共鉴定雌雄蛹200头,成虫200头,准确率达100%。这些结果将为米缟螟的性行为学、化学生态学以及资源昆虫开发利用等方面的研究奠定基础。