云南部分地区大绒鼠种群遗传多样性与遗传分化研究

大绒鼠Eothenomys miletus是云南省野鼠鼠疫疫源地的主要宿主动物之一。本研究对云南部分地区大绒鼠线粒体细胞色素b(cyt b)基因的部分序列进行分析,探讨云南不同地理种群大绒鼠的遗传多样性和遗传分化。共获得7个采样点140只大绒鼠的cyt b基因序列,共发现变异位点109个,定义51个单倍型,所有种群均为高单倍型多样性和低核苷酸多样性模式,整体平均单倍型多样性为0.977±0.004,核苷酸多样性为0.018 45±0.001 78。基于类平均法构建的分子系统发育树和基于中连法构建的单倍型网络关系图显示出相同的结果,大绒鼠种群分化为4个地域性分支。基因流数据显示多数地理种群间基因交流贫乏,分子变异分析表明,种群间的遗传差异(84.29%)明显高于种群内(15.71%),遗传分化显著。IBD分析表明,大绒鼠的遗传分化与地理距离呈正相关(r=0.699 7,P0.05),说明地理距离隔离是大绒鼠种群分化的重要原因之一。大绒鼠的这种遗传多样性与种群遗传分化,可能与其地下洞道生活、扩散能力弱、地理距离隔离等因素有关。本研究为大绒鼠的分子生态学研究提供了参考依据。     

中哈边境区域黑腿星翅蝗遗传多样性与遗传分化研究

【目的】为了明确中哈边境区域黑腿星翅蝗Calliptamus barbarus Costa的遗传多样性,阐明其区域性发生的遗传机制。【方法】测定中哈边境新疆边境区域阿勒泰、塔城、博乐、伊犁等和哈萨克斯坦境内黑腿星翅蝗5个种群100个体的COⅠ基因全长序列(1 540 bp),利用DnaSP 5.0,MEGA 6.0和Arlequin3.5等软件分析种群遗传多样性与分化情况。【结果】共获得100条COⅠ基因序列,在5个地理种群中发现122个变异位点,包含69个单倍型,其中1个单倍型为4个种群所共享。种群间的遗传距离范围为0.001-0.086。总群体单倍型多样性指数H_d=0.987,各地理种群单倍型多样性介于0.973-0.995之间,总群体核苷酸多样性P_i=0.008 4,总群体的遗传分化系数G_(st)=0.004 5,总群体固定系数F_(st)=0.014 2,总基因流N_m=4.61。中性检验(Tajima's D=-1.553 3,P0.05;Fu’s Fs=-3.732 4,P0.05)结果表明中哈边境黑腿星翅蝗种群在较近的历史上没有出现群体扩张。单倍型网络图和NJ单倍型系统树结果一致,博乐种群与其他种群分化较明显,AMOVA分子变异分析结果表明中哈边境不同地理种群黑腿星翅蝗的遗传分化主要来自种群内部(98.56%),种群间变异水平很低(1.44%)。不同地理种群的遗传距离与地理距离间的相关性不显著。各单倍型散布在不同地理种群中,未形成明显的系统地理结构。【结论】中哈边境区域不同地理种群黑腿星翅蝗基因交流频繁,遗传分化程度低。     

基于线粒体COⅡ基因序列的中华蜜蜂地理种群的遗传多样性研究

通过分析我国20个不同地理种群中蜂线粒体COⅡ序列的变异,对中华蜜蜂群体遗传分化程度遗传多样性进行全面研究。结果表明:COⅡ基因部分序列中共发现16个变异位点和18个单倍型,核苷酸差异数的平均值为0.939,核苷酸分歧度(Dxy)在0.1%~0.965%之间变化,核苷酸遗传距离为-0.007%~1.489%。总种群的Fst为0.4978,差异均极显著(P0.001)。研究结果显示种群总体单倍型多样性较为丰富,种群间核苷酸分歧度差异很大。20个东方蜜蜂不同地理种群间存在显著的遗传分化。     

基于线粒体COI序列变异的高原鼢鼠种群遗传分化研究

基于线粒体COI基因序列对高原鼢鼠7个地理种群的遗传多样性、遗传分化和基因流进行分析,158条COI基因序列中发现了570个变异位点,界定了54个COI单倍型。总体单倍型多样性指数Hd为0.911,种群单倍型多样性在0.278—0.964之间,高原鼢鼠的种群遗传多样性较高。群体总的遗传分化系数Fst为0.611,群体间的基因流Nm在-0.020—3.700之间,部分地理种群之间的遗传分化程度高,基因流弱。AMOVA分子变异分析显示,高原鼢鼠的遗传变异主要来自种群之间(77.56%),种群内部的遗传变异较低(22.44%)。中性检验(P0.01)与错配分析显示高原鼢鼠种群经历了群体扩张事件。IBD未检测到地理距离与种群间遗传距离的显著性相关关系(R2=0.124,P=0.118)。综合分析认为,高原鼢鼠不同地理种群间遗传分化的原因除地理隔离外,还与迁移扩散方式、进化过程和其他环境因素有关。     

基于线粒体COI基因序列的内蒙古沙葱萤叶甲种群遗传多样性及遗传分化

沙葱萤叶甲Galeruca daurica(Joannis)是一种近年来在内蒙古草原上猖獗发生为害的新害虫,为明确内蒙古沙葱萤叶甲不同地理种群间的遗传多样性及遗传分化,本研究测定了内蒙古8个地理种群的197头沙葱萤叶甲的线粒体COI基因417 bp序列,利用Dna SP 5.0、Arlequin 3.11和Network 4.6.1.0等软件对沙葱萤叶甲种群间的遗传多样性、遗传分化、分子变异等进行分析。在所分析的197个COI序列中,共有43个变异位点和62个单倍型,其中2个单倍型为各种群共享,每个种群具有3个及以上的独享单倍型。总种群的单倍型多态度指数Hd为0.9466,不同地理种群单倍型多态度Hd在0.8889-0.9600。总群体固定系数Fst为0.0315,基因流Nm为15.37。中性检验结果不显著(Tajima's D=-1.6300,0.10P0.05),说明内蒙古沙葱萤叶甲在较近的历史时期内没有出现种群扩张现象。分子变异分析(AMOVA)结果表明,沙葱萤叶甲的遗传分化主要来自种群内部(96.34%),而种群间未发生明显的遗传分化。各种群间遗传分化程度与地理距离无显著相关性。不同地理种群的单倍型在中介网络图上散布在不同的分布群中,未形成明显的地理分布格局。结果表明沙葱萤叶甲不同地理种群遗传多样性高,基因交流程度高,遗传分化不明显且与地理距离无显著相关性。     

迁飞性昆虫甜菜夜蛾遗传分化研究

【目的】甜菜夜蛾Spodoptera exigua逐渐成为世界性重要害虫,为明确甜菜夜蛾不同地理种群的遗传分化及遗传多样性,本研究探讨其在中国的种群遗传变异。【方法】测定了采自15个地理种群154个甜菜夜蛾的线粒体COⅠ基因的547 bp序列,利用DnaSP 5.0、Arlequin 3.5.1.2等软件对甜菜夜蛾不同种群间的遗传多样性、遗传分化及分子变异进行分析,并建立单倍型系统进化树。【结果】在所分析的154个COⅠ序列中,共检测出5个单倍型,其中H1为各种群所共享种群内遗传多样性较低(Hd=0.172±0.041,Pi=0.00077±0.0002),种群内遗传分化相对较大(FST=0.3182),基因流水平较高(Nm=1.071)。中性检验结果不显著(Tajima’s D=-1.278,P0.10,Fu’s Fs=-1.660,P0.10),说明中国地区甜菜夜蛾在较近的历史时期内没有出现种群扩张现象。分子变异分析(AMOVA)结果表明,甜菜夜蛾遗传变异主要来自种群内部(68.18%),而种群间未发生明显的遗传分化。根据各地理种群的单倍型建立的系统发育树表明,各单倍型散布在不同的地理种群中,无明显的地理分布格局。【结论】甜菜夜蛾不同种群的遗传距离与地理距离间无显著线性相关性,不同种群间的基因交流不受地理距离的影响。这些数据表明,除了遗传因素之外,不同因素的组合,例如地理距离、环境条件和生理行为,可能在甜菜夜蛾种群内和之间形成变异中起重要作用。     

基于线粒体COⅡ基因的大豆蚜不同地理种群遗传分化研究

【目的】大豆蚜Aphis glycines逐渐成为栽培大豆上的世界性重要害虫,为明确大豆蚜不同地理种群的遗传分化及遗传多样性,本研究探讨其在中国的种群遗传变异。【方法】测定了采自7个省共14个地理种群339头大豆蚜的线粒体COⅡ基因的673 bp序列,利用Dna SP 5.0、Arlequin 3.5.1.2、Network4.6.1.3等软件对地理种群间的大豆蚜的遗传多样性、遗传分化程度及分子变异进行分析,并建立了单倍型网络图及单倍型系统进化树。【结果】在所分析的339个COⅡ序列中,共检测出7个单倍型,其中H1为各种群所共享。种群内遗传多样性较低(Hd=0.479±0.030,Pi=0.00166±0.00018),种群内遗传分化相对较大(Fst=0.1985),基因流水平较高(Nm=2.019)。中性检验结果不显著(Tajima’s D=﹣0.931,P>0.10,Fu’s Fs=0.220,P>0.10),说明中国地区大豆蚜在较近的历史时期内没有出现种群扩张现象。分子变异分析(AMOVA)结果表明,大豆蚜遗传变异主要来自种群内部(80.15%),而种群间未发生明显的遗传分化。根据各地理种群的单倍型建立的系统发育树、单倍型网络图表明,各单倍型散布在不同的地理种群中,无明显的地理分布格局。【结论】大豆蚜不同地理种群的遗传多样性不高,各种群的遗传距离与地理距离之间没有显著线性相关性,各种群间的基因交流并未受到地理距离的影响。     

基于线粒体COⅡ基因的中国大豆食心虫不同地理种群遗传分化分析

【目的】大豆食心虫Leguminivora glycinivorella(Matsumura)是我国大豆Glycina max最重要的蛀荚害虫。本研究旨在探讨中国大豆食心虫不同地理种群的遗传分化情况。【方法】通过测定大豆食心虫19个地理种群208头老熟幼虫的线粒体COⅡ基因序列(723 bp),利用MEGA6.0和Dna SP 5.0等软件分析大豆食心虫不同地理种群的基因序列变异和遗传分化情况。【结果】在19个大豆食心虫地理种群中共获得208条COⅡ基因序列,发现61个变异位点,包含57个单倍型。总群体单倍型多样度Hd=0.8522,各地理种群单倍型多样度范围在0.1818~0.9546间,总群体的固定系数Fst=0.4619,总遗传分化系数Gst=0.1186,总基因流Nm=0.29。总群体Tajima’s D检验结果不显著,表明大豆食心虫在较近的历史时期未出现群体扩张现象,群体大小稳定。单倍型网络图、ML单倍型系统树及采用不同地理种群间遗传距离构建的UPGMA系统发育树均显示,贵阳(GY)和都安(DA)种群与其余地理种群分化明显。AMOVA分子变异分析结果显示,东北组、西北组、黄淮组、华南组及西南组种群组间所占总变异的比例大于组内。【结论】大豆食心虫不同地理种群间基因交流较少,整体的遗传多样性和遗传分化程度较高,遗传分化主要来自地理种群组间,且地理距离是影响不同地理种群间遗传距离的重要因素。     

基于mtCOI基因序列的中国葱斑潜蝇不同地理种群遗传多样性分析

【目的】葱斑潜蝇Liriomyza chinensis是葱蒜类蔬菜上的重要经济害虫,在我国广泛分布。本研究旨在阐析中国葱斑潜蝇的地理种群遗传分化。【方法】以中国8省12个不同地理种群的253头葱斑潜蝇为样本,测定其mtCOI基因序列;依据获得的mtCOI基因序列,利用MEGA7.0, DnaSP 6.1和Arlequin 3.5等软件对葱斑潜蝇地理种群的遗传多样性、基因流水平和遗传变异等进行分析。【结果】在253头个体的759 bp mtCOI基因片段中,获得13个单倍型,各单倍型间K2P遗传距离均小于0.02;其中单倍型Hap1为12个地理种群所共享,总发生频率高达81.82%。总群体单倍型多样性较低(H_d=0.327),核苷酸多样性(Pi)为0.00159,核苷酸平均差异数(K)为1.21011;葱斑潜蝇总群体遗传分化程度中等(F_ST=0.06971),基因交流较充分(Nm=3.33629)。分子方差分析(AMOVA)结果表明,遗传变异来源为群体内;总群体Tajima’s D检验值为显著负值;Mantel检测结果说明种群间的遗传距离与地理距离没有相关性。【结论】中国葱斑潜蝇地理种群的遗传多样性较低,基因交流较充分,遗传分化程度中等,且地理距离并不影响其遗传分化程度;葱斑潜蝇总群体在较近的历史时期未经历明显的种群扩张和种群增长。     

基于线粒体COI基因序列的大豆食心虫中国东北地理种群遗传多样性分析

【目的】大豆食心虫Leguminivora glycinivorella (Matsumura)是一种危害大豆的主要害虫,在中国北方地区危害较重。本研究旨在探讨大豆食心虫在中国东北不同地理种群间的遗传变异。【方法】测定了10个不同地理种群153个个体的线粒体细胞色素氧化酶亚基Ⅰ (mtCOI)基因的657 bp序列,利用DnaSP 5. 0和Arlequin 3. 5. 1. 2等软件对大豆食心虫种群间的遗传多样性、基因流水平和分子变异进行分析。【结果】结果表明：10个地理种群间的COI基因共有36个变异位点和17个单倍型,其中1个单倍型为10个种群所共享。总种群的单倍型多样性指数Hd为0.456,各地理种群单倍型多样度范围在0~0.634之间。总群体的固定系数Fst为0.12545,遗传分化系数Gst为0.06326,总基因流Nm为3.49,且各种群间的基因流均大于1,种群间基因交流的水平较高。【结论】大豆食心虫种群内遗传多样性水平处于中低等水平。总群体和各种群的Tajima’s D检验结果皆不显著,说明中国东北地区大豆食心虫在较近的历史时期内没有出现种群扩张现象。AMOVA分子变异分析结果表明,大豆食心虫的遗传分化主要来自种群内部,而种群间未发生明显的遗传分化。各地理种群的单倍型在系统发育树上和中介网络图上散布在不同的分布群中,缺乏明显的地理分布格局。各种群的遗传距离与地理距离之间没有显著线性相关性,种群间的基因交流并未受到地理距离的影响。     

Therps16 intron and the phylogeny of theRubioideae (Rubiaceae)

The phylogeny of the subfamilyRubioideae (Rubiaceae) was estimated from sequence variation in therps16 intron (cpDNA) in 143 ingroup and 5 outgroup taxa. The analysis largely confirms a recent one based onrbcL sequences, but branch support is often much stronger. Three of the traditional subfamilies are supported,Rubioideae, Cinchonoideae s. str., andIxoroideae s. l. while there is no support forAntirheoideae. TheRubioideae are the sister group of all otherRubiaceae and comprise the tribesAnthospermeae, Coccocypseleae, Cruckshanksieae, Coussareeae, Gaertnereae, Hedyotideae, Knoxieae, Morindeae, Ophiorrhizeae, Paederieae, Pauridiantheae, Perameae, Psychotrieae, Rubieae, Spermacoceae, Theligoneae, andUrophylleae. TheHamelieae andHillieae belong to theCinchonoideae. Rachicallis andSiemensia should be transferred from theHedyotideae to theCinchonoideae. ThePauridiantheae, Urophylleae, Ophiorrhizeae, andRaritebe form the basalmost subclade of theRubioideae. The second basalmost clade consists of the generaLasianthus andPerama. The third basalmost clade consists of the tribesCoussareeae, Coccocypseleae andCruckshanksieae, and the generaDeclieuxia andHindsia. The tribesKnoxieae, Anthospermeae, Argostemmateae, Paederieae, Theligoneae, Rubieae, Hedyotideae, andSpermacoceae are members of one clade. TheKnoxieae are monophyletic ifOtiophora, Otomeria, andPentas are included. The tribeAnthospermeae is supported as monophyletic, but its subtribes are not. ThePaederieae, together withTheligonum, form a paraphyletic grade basal to theRubieae. TheHedyotideae, includingSchismatoclada, form a grade at the base of theSpermacoceae. TheGaertnereae are monophyletic and distinct from thePsychotrieae. TheMorindeae are monophyletic and includeDamnacanthus andMitchella. Schradera is the sister group of theMorindeae. ThePsychotrieae are monophyletic when theGaertnereae, Lasianthus, andDeclieuxia are excluded. The recognition of a subtribeHydnophytineae leaves the rest of thePsychotrieae paraphyletic.Psychotria is paraphyletic with respect to all other genera of the tribe. Approximately 50 genera are here classified for the first time based on molecular data.     

A review of selected South- and East Asian Tanytarsus v.d. Wulp (Diptera: Chironomidae)

Male and pupa of Tanytarsus calorifontis sp. n. are described. Diagnoses, hypopygium drawings for the examined species and distribution are given for: Tanytarsus akantertiusSasa & Kamimura, T. angulatus Kawai, T. atagoensis Tokunaga,T. bathophilusKieffer, T. boninensis Tokunaga, T. formosae (Kieffer), T. formosanusKieffer, T. infundibulusChaudhuri & Datta, T. kikuchiiSasa, Kawai & Ueno, T. konishii Sasa & Kawai, T. mcmillani Freeman, T. mendax Kieffer, T. miyakobrevis Sasa & Hasegawa, T. monstrosus Chaudhuri et al., T. ogasaquartus Sasa & Suzuki, T. ogasatertius Sasa & Suzuki, T. okuboi Sasa & Kikuchi, T. oscillans Johannsen, T. ovatus Johannsen, T. oyamai Sasa, T. pollexus Chaudhuri & Datta, T. shouautumnalis Sasa, T. shoudigitatus Sasa, T. takahashii Kawai & Sasa, T. tamaundecimus Sasa, T. tonebeceus Sasa & Tanaka, T. tusimatneousSasa & Suzuki, T. unagiseptimus Sasa, T. uraiensis Tokunaga, T. yakuheiusSasa & Suzuki and T. yunosecundus Sasa. Tanytarsus ikiefeus Sasa & Suzuki is a new junior synonym of T. konishii. Tanytarsus ikifegeus Sasa & Suzuki, T. miyakoflavus Sasa & Hasegawa,T. oyabepallidus Sasa, Kawai & Ueno, T. simantoopeus Sasa et al. and T. tusimatheius Sasa & Suzuki are new junior synonyms of T. okuboi. Tanytarsus nippogregarius Sasa & Kamimura is a new junior synonym of T. bathophilus. Tanytarsus cultellus Chaudhuri & Datta and T. sibafegeus Sasa et al. are new junior synonyms of T. oscillans.Tanytarsus insulus (Guha et al., 1985) is a new junior synonym of T. ovatus. Tanytarsus sakishimanus Sasa & Hasegawa, T. vinculus Chaudhuri et al., T. parvistylus Chaudhuri & Datta, T. fusciabdominalis Guha et al. and T. euformosanus Kikuchi & Sasa are new junior synonyms of T. formosanus. Tanytarsus tsutaprimus Sasa, T. tokarajekeus Sasa & Suzuki, T. tusimatlemeus Sasa & Suzuki, T. tusimatopeus Sasa & Suzuki and T. yakugeheuus Sasa & Suzuki are new junior synonyms of T. shouautumnalis. Tanytarsus togasiroidus Sasa & Okazawa is a new junior synonym of T. shoudigitatus. Tanytarsus tusimatjekeus Sasa & Suzuki and T. tusimatkeleus Sasa & Suzuki are new junior synonyms of T. akantertius, and Tanytarsus tusimatpequeus Sasa & Suzuki is a new junior synonym of T. tusimatneous. Virgatanytarsus toganiveus (Sasa & Okazawa), Cladotanytarsus utonaiquartus (Sasa) and Zavrelia tusimatijeus (Sasa & Suzuki), all previously placed in Tanytarsus, are new combinations.     

Influence of six aphid prey species on development and reproduction of a ladybird beetle, Coccinella septempunctata

Pre-imaginal development, immaturesurvival, and reproduction of a ladybird, Coccinella septempunctata Linnaeus, werestudied in response to six aphid species, Aphiscraccivora Koch, Aphis gossypii Glover,Aphis nerii Boyer de Fonscolombe,Lipaphis erysimi (Kaltenbach), Myzuspersicae (Sulzer) and Uroleuconcompositae (Theobald) to quantify theirrelative suitability as prey. Pre-adultdevelopment was shortest (13.93 ± 0.12 days)when fed on L. erysimi and longest(22.85 ± 0.10 days) on A. nerii. Immaturesurvival, adult emergence, growth index,relative growth rate, development rate, maleand female longevity, oviposition period,fecundity and hatching percent were maximal, i.e. 73.47 ± 0.89%, 90.07 ± 1.43%,8.62 ± 0.23, 1.52 ± 0.02, 0.07,81.10 ± 1.26 days, 85.70 ± 1.45 days,69.80 ± 1.32 days, 1764.10 ± 8.46,and 87.88 ± 1.05, respectively when C.septempunctata were fed on L. erysimi.The same parameters were minimal, i.e.43.86 ± 1.33%, 71.65 ± 2.75%,2.02 ± 0.08, 0.49 ± 0.02, 0.04,44.40 ± 1.39 days, 53.50 ± 1.00 days,16.40 ± 0.60 days, 203.20 ± 11.83, and48.68 ± 2.06, respectively on A. nerii. Theweights of different ladybird life stages weremaximal after feeding on L. erysimi and minimalon A. nerii. Regression analyses of thedata revealed linear relationships betweendevelopment rate and weight of adult; dailyprey consumption and relative growth rate; logweight of adult male and female; and longevityand fecundity of female. On the basis of thesefindings, the order of suitability of aphidspecies for C. septempunctata is L. erysimi >M. persicae > A. craccivora > A. gossypii >U. compositae > A. nerii. Thus, the presentinformation can be utilized for the massrearing of C. septempunctata by supplyingthe best food and can also help in theprediction of the relative abundance of theladybird on different aphid infestations in thefields.     

Comparative Virulotyping of Salmonella typhi and Salmonella enteritidis

Members of Salmonella enterica are important foodborne pathogens of significant public health concern worldwide. This study aimed to determine a range of virulence genes among typhoidal (S. typhi) and non-typhoidal (S. enteritidis) strains isolated from different geographical regions and different years. A total of 87 S. typhi and 94 S. enteritidis strains were tested for presence of 22 virulence genes by employing multiplex PCR and the genetic relatedness of these strains was further characterized by REP-PCR. In S. typhi, invA, prgH, sifA, spiC, sopB, iroN, sitC, misL, pipD, cdtB, and orfL were present in all the strains, while sopE, agfC, agfA, sefC, mgtC, and sefD were present in 98.8, 97.7, 90.8, 87.4, 87.4 and 17.2 %, of the strains, respectively. No lpfA, lpfC, pefA, spvB, or spvC was detected. Meanwhile, in S. enteritidis, 15 genes, agfA, agfC, invA, lpfA, lpfC, sefD, prgH, spiC, sopB, sopE, iroN, sitC, misL, pipD, and orfL were found in all S. enteritidis strains 100 %, followed by sifA and spvC 98.9 %, pefA, spvB and mgtC 97.8 %, and sefC 90.4 %. cdtB was absent from all S. enteritidis strains tested. REP-PCR subtyped S. typhi strains into 18 REP-types and concurred with the virulotyping results in grouping the strains, while in S. enteritidis, REP-PCR subtyped the strains into eight profiles and they were poorly distinguishable between human and animal origins. The study showed that S. typhi and S. enteritidis contain a range of virulence factors associated with pathogenesis. Virulotyping is a rapid screening method to identify and profile virulence genes in Salmonella strains, and improve an understanding of potential risk for human and animal infections.     

Use of a single injection of long-acting recombinant bovine FSH to superovulate Holstein heifers: A preliminary study

Our objective was to compare several experimental preparations of a single injection of long-acting recombinant bovine FSH (rbFSH; types A and B) to a porcine pituitary-derived FSH (Folltropin) to superovulate Holstein dairy heifers. Nonlactating, nonpregnant virgin Holstein heifers (n = 56) aged 12 to 15 months were randomly assigned to one of four superstimulatory treatments. Beginning at a random stage of the estrous cycle, all follicles greater than 5 mm were aspirated. Thirty-six hours later, heifers received an intravaginal P4 device and superstimulatory treatments were initiated. Treatments were (1) 300 mg of pituitary-derived FSH (Folltropin) administered in eight decreasing doses over a period of 3.5 days; (2) a single injection of 50 μg of A-rbFSH; (3) a single injection of 100 μg of A-rbFSH; and (4) a single injection of 50 μg of B-rbFSH. All heifers received 25 mg PGF_2α at 48 and 72 hours after the insertion of P4 device. At 84 hours after insertion, P4 devices were removed, and ovulation was induced 24 hours later with hCG (2500 IU). Heifers were inseminated at 12 and 24 hours after hCG treatment. The number of ovulatory follicles was greatest for heifers treated with Folltropin and B50-rbFSH, least for heifers treated with A50-rbFSH, and was intermediate for heifers treated with A100-rbFSH (25.7 ± 3.2, 18.9 ± 3.2, 5.9 ± 0.9, and 16.6 ± 3.1, respectively; P < 0.001). The number of corpora lutea was greatest for heifers treated with Folltropin, B50-rbFSH, and A100-rbFSH, and least for heifers treated with A50-rbFSH (19.1 ± 2.4, 16.1 ± 3.0, 15.9 ± 2.9, and 2.6 ± 0.9, respectively; P < 0.001). The number of good-quality embryos differed among treatments and was greatest for heifers treated with B50-rbFSH, Folltropin, and A100-rbFSH and least for heifers treated with A50-rbFSH (7.6 ± 2.4, 6.5 ± 1.7, 4.3 ± 1.5, and 0.8 ± 0.5, respectively; P < 0.001). In conclusion, a single injection of a preparation of long-acting rbFSH (either 100 μg of A-rbFSH or 50 μg of B-rbFSH but not 50 μg of A-rbFSH) produced similar superovulatory responses resulting in the production of good-quality embryos when compared with a pituitary-derived FSH preparation administered twice daily for 4 days. More studies using different types of cattle and different doses of rbFSH are needed to confirm the findings reported in this preliminary study.     

Collection and characterization of semen in Mithun (Bos frontalis) bulls

The objective of this study was to collect semen from semiwild Mithun (Bos frontalis) bulls using an artificial vagina (AV) and to determine semen characteristics. Collection of semen with an AV was attempted in five Mithun bulls using both anestrous and estrous Mithun females. No Mithun bull mounted an anestrous female Mithun during 60 trials, but satisfactory mounting, including extension of the penis, occurred in 25 trials with estrous Mithun females. In 15 of these trials, semen was successfully collected in an AV with an internal temperature of 42 to 46 °C. However, in 10 trials with an AV with an internal temperature of 36 to 40 °C, semen was not collected. Mean (± SEM) intervals to first mount and to ejaculation in the AV were 27.9 ± 3.6 sec and 113.8 ± 6.6 sec, respectively. Semen volume and pH were 3.1 ± 0.35 mL and 6.59 ± 0.04, and mean mass activity (scale, 0 to 4), initial sperm motility, live sperm count, sperm concentration, total number of sperm in the ejaculate, and overall sperm length were 2.2 ± 0.3, 78.6 ± 2.6%, 80.7 ± 2.2%, 710.8 ± 66.8 × 10⁶/mL, 2114 ± 364.4 sperm, and 67.9 ± 0.6 μm, respectively. The proportion of morphologically normal sperm was 80.6 ± 0.2%, whereas the proportion with a morphologically abnormal head, midpiece, tail, and acrosome were 4.2 ± 0.4%, 1.6 ± 0.5%, 6.1 ± 1.1%, and 7.1 ± 0.9%, respectively. The mean incidence of tail-less heads and proximal and distal protoplasmic droplets were 0.5 ± 0.1%, 0.3 ± 0.2%, and 2.4 ± 0.3%, respectively. In conclusion, we successfully collected semen from semiwild Mithun bulls with an AV maintained at 42 to 46 °C, and overall, the semen was within the normal range of that collected from fertile domestic bulls.     

A study of the distribution of chitosan onto and within a paper sheet using a fluorescent chitosan derivative

Acidic glutaraldehyde (Gh) crosslinked chitosan (ChGhH) when deprotonated the biopolymer (ChGh) presents high content of free amino groups. These modified biopolymers are comparable to epichlorohydrin (Ep) crosslinked (ChEp). C/N molar ratio of 6.1 for chitosan increases to 7.3, 7.5 and 7.1 for ChGhH, ChGh and ChEp. The effectiveness of the carbon-6 hydroxyl group in interconnecting chitosan units was supported by IR and ¹³CNMR, where Ep promotes increase in crystallinity. Copper uptake gave the order Ch > ChGh > ChGhH > ChEp, as: 1.35 ± 0.06, 1.30 ± 0.05, 1.05 ± 0.07 and 0.96 ± 0.22 mmol g⁻¹, reflecting the availability of nitrogen basic centers in adsorbing. The favorable thermodynamic data of adsorption through calorimetric titration gave exothermic enthalpic values: −28.98 ± 0.05, −6.68 ± 0.04, −6.13 ± 0.07 and −0.65 ± 0.23 kJ mol⁻¹ for Ch, ChGh, ChGhH and ChEp. Free Gibbs energy reflected spontaneity of interactions and, with the exception of chitosan, the entropic values are positive.     

Checklist of the helminth parasites of the genus Profundulus Hubbs, 1924 (Cyprinodontiformes,Profundulidae), an endemic family of freshwater fishes in Middle-America

From December 2012 to November 2014, 267 fish belonging to the family Profundulidae (representing nine of the 11 species of the genus Profundulus) were collected in 26 localities of Middle-America, across southern Mexico, Guatemala, and Honduras, comprising the distribution range of the genus, and analyzed for helminth parasites. Additionally, a database with all ten available published accounts of the helminth parasite fauna of this genus (the only genus within the family) was assembled. Based on both sources of information, a checklist containing all the records was compiled as a tool to address future questions in the areas of evolutionary biology, biogeography, ecology and phylogeography of this host-parasite association. The helminth parasite fauna of this fish group consists of 20 nominal species, classified in 17 genera and 14 families. It includes six species of adult digeneans, five metacercariae, two monogeneans, one adult cestode, three adult nematodes and three larval nematodes. The profundulid fishes are parasitized by a specialized group of helminth species (e.g. Paracreptotrema blancoi sensu Salgado-Maldonado et al. (2011b), Paracreptotrema profundulusi Salgado-Maldonado, Caspeta-Mandujano & Martínez Ramírez, 2011, Phyllodistomum spinopapillatum Pérez-Ponce de León, Pinacho-Pinacho, Mendoza-Garfias & García-Varela, 2015, Spinitectus humbertoi Mandujano-Caspeta & Moravec, 2000, Spinitectus mariaisabelae Caspeta-Mandujano Cabañas-Carranza & Salgado-Maldonado, 2007 and Rhabdochona salgadoi Mandujano-Caspeta & Moravec, 2000), representing the core helminth fauna that are not shared with other Middle-American fish species.