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1.
通过以鸡白痢沙门菌为宿主菌,采用双层琼脂平板法从鸡场粪样中分离噬菌体。结果获得1株噬菌体,并对其进行形态学的鉴定,同时测定噬菌体对宿主的敏感性。经透射电镜观察发现该噬菌体无明显尾部,头部大致呈六角形,直径约为27.3 nm,其噬菌斑清晰透亮,周围无晕环,具有宽噬菌谱的特性,不仅可强裂解宿主菌,还可裂解1株大肠杆菌。获得的这株噬菌体可用于家禽生产中鸡白痢沙门菌感染的防治。  相似文献   

2.
为了分离一株裂解性鸡白痢沙门氏菌噬菌体,对抗多重耐药细菌,试验从健康鸡粪便中分离富集噬菌体,采用双层琼脂平板法纯化、增殖,超速离心法浓缩,利用透射电镜观察噬菌体形态,通过提取噬菌体基因组、酶切、电泳确定核酸类型,同时测定噬菌体的宿主谱、最佳感染复数、一步生长曲线、酸碱及热稳定性等生物学特性。结果表明:从鸡粪中分离得到1株裂解性噬菌体,命名为PSP2-22。该噬菌体头部直径为(60±5)nm,尾部长为(130±5)nm,为长尾噬菌体科;核酸类型为双链DNA。该噬菌体不仅能裂解鸡白痢沙门氏菌,还可裂解鼠伤寒沙门氏菌和奇异变形杆菌;最佳感染复数为0.1;平均裂解量约为136 pfu/cell,效价最高可达9.83 lg pfu/mL;在pH值6~11时相对稳定,70℃作用60 min之后仍具有一定活性。说明噬菌体PSP2-22是一株对不同温度和pH值有较强适应能力的双链DNA噬菌体,宿主谱相对较宽,具有潜在的应用价值。  相似文献   

3.
【目的】探究大肠杆菌噬菌体vB_EcoP_GN07的生物学特性和全基因组特征,为开发噬菌体“鸡尾酒”制剂防治致病性大肠杆菌病提供生物学材料。【方法】采用双层平板法从广西南宁某养鸡场分离到1株大肠杆菌裂解性噬菌体vB_EcoP_GN07;通过透射电镜观察、宿主谱测定、pH和温度稳定性评估、最佳感染复数(MOI)测定、一步生长曲线测定、全基因组测序等方法了解噬菌体的形态学、生物学特性和全基因组特性。【结果】透射电镜显示,vB_EcoP_GN07头部直径为45 nm±5 nm,尾部长约25 nm,属于短尾噬菌体科,宿主特异性强;在4~60℃可以保持效价稳定,在pH 3.0~11.0可以保持噬菌体活性;该噬菌体的最佳MOI为10-2;一步生长曲线显示该噬菌体潜伏期为60 min,裂解量为560 PFU/cell;测序结果显示其属于双链线型DNA,长度为70 120 bp, GC含量为42.8%;噬菌体vB_EcoP_GN07共有81个开放阅读框,其中23个为已知功能蛋白。经BLASTN比对显示,vB_EcoP_GN07与NCBI数据库中其他噬菌体相似性较低,符合N4-li...  相似文献   

4.
【目的】分离有效裂解沙门氏菌的噬菌体,为探索新的沙门氏菌防控方法提供理论依据和参考。【方法】以鸡白痢沙门氏菌X-1014为宿主菌,通过双层琼脂平板法从污水中分离并纯化出噬菌体。采用噬菌斑与电子显微镜观察、宿主谱测定、生物学特性测试、体外抑菌试验及噬菌体的全基因组测序分析方法评估该噬菌体。【结果】分离纯化出1株能裂解沙门氏菌和大肠杆菌的裂解性噬菌体,命名为ZH5(GenBank登录号:OM864357.1)。双层平板法结果显示,噬菌斑透亮且边缘清晰。电镜观察显示,噬菌体ZH5头部呈二十面体、头部直经约50 nm,尾部长度约20 nm。宿主谱分析结果显示,噬菌体ZH5能裂解3株沙门氏菌和1株大肠杆菌。生物学特性结果显示,最佳感染复数为10,潜伏期和爆发期分别为20和60 min,裂解量约为139 PFU/cell;酸碱耐受范围为pH 4.0~10.0,且在温度为4~50℃的环境中能存活30 min。体外抑菌结果显示,当感染复数为10时,噬菌体ZH5对沙门氏菌X-1014和CVCC 1806抑制效果最佳。全基因组分析结果显示,该噬菌体全长为42 949 bp, GC含量为51.41%,其中已...  相似文献   

5.
本研究旨在通过双层平板法从鸡粪中分离鼠伤寒沙门氏菌烈性噬菌体,并通过电镜观察、热稳定性、pH稳定性、最佳感染复数、一步生长曲线和在小鼠上的防制效果试验对噬菌体进行评估。结果显示,从鸡粪中成功分离出一株鼠伤寒沙门氏菌噬菌体(φBLCC8-0050-3),该噬菌体有一个正多面体立体对称的头部,头径直径约65 nm,无尾部结构,应属于盖噬菌体科(Tectiviridae)烈性噬菌体;噬菌斑成圆形空斑,周围有晕环,直径为2~3 mm;温度耐受范围是30~50℃;pH耐受范围为3~10。当感染复数(multiplicity of infection,MOI)为0.0001时,子代噬菌体滴度最高;按照一步生长曲线结果,潜伏期为20 min左右,爆发时间为180 min左右,爆发量为43 333 PFU/cell;从噬菌体对鼠伤寒沙门氏菌的防制效果来看,处理14 d后,阳性对照组小鼠存活率为0,噬菌体预防组小鼠存活率为90%,噬菌体治疗组小鼠存活率为70%。说明与阳性对照组相比,预防组和治疗组都对小鼠抵抗沙门氏菌感染有一定的作用。结果表明,φBLCC8-0050-3是一株具有良好应用前景的噬菌体,可作为一种生物抑菌剂用于鼠伤寒沙门氏菌的防制。  相似文献   

6.
本研究从养殖场采集的污水样品中分离到宽噬菌谱肠出血性大肠杆菌烈性噬菌体V_EcoM_C1,并进行了噬菌斑、噬菌谱、形态学(透射电镜)、遗传物质、酸碱及热稳定性、一步生长曲线等生物学特性及控制养殖环境中的肠出血性大肠杆菌的污染研究。结果表明:噬菌体V_EcoM_C1呈现出典型裂解性噬菌体特征,空斑透亮,无晕环,空斑边缘整齐清晰;噬菌体V_EcoM_C1噬菌斑直径为1.0~1.5 mm;噬菌体V_EcoM_C1头部椭圆形,长径约86 nm,横径约54 nm;噬菌体V_EcoM_C1具有可收缩性尾,尾长约76 nm,直径约10 nm,头部与尾部被颈圈隔开;噬菌体V_EcoM_C1可裂解5种不同血清型肠出血性大肠杆菌,且噬菌斑透亮,具有较宽噬菌谱;噬菌体V_EcoM_C1在30℃~50℃保持高活性;噬菌体V_EcoM_C1在pH5~10稳定;噬菌体V_EcoM_C1感染宿主菌的潜伏期约10 min,爆发持续时间约60 min,平均爆发量为26,具有较高的裂解活性;噬菌体V_EcoM_C1可以控制养殖环境中的肠出血性大肠杆菌证明该噬菌体可以有效杀灭养殖环境(奶牛料槽表面)中的肠出血性大肠杆菌。综上所述,V_EcoM_C1为1株宽噬菌谱肠出血性大肠杆菌烈性噬菌体,可用于肠出血性大肠杆菌噬菌体制剂的研发。  相似文献   

7.
为分离到能有效裂解黑山羊源大肠杆菌的烈性噬菌体,本课题采集不同来源的新鲜粪便样品并经目的菌富集后,结合双层琼脂平板法进行噬菌体分离纯化,随后制作透射电镜样品进行其外形特征观察;对获得的噬菌体进行裂解谱、最佳感染复数和生长曲线分析;该噬菌体对温度、pH和紫外线的敏感性测定;该噬菌体全基因组测定与分析。结果表明,从黑山羊粪便中分离出的一株烈性噬菌体PLYYY01,在大肠杆菌菌苔形成1~2 mm的透亮噬菌斑;通过透射电镜观察,该噬菌体头部直径在60nm左右,尾部长度为100nm左右;该噬菌体有较宽的裂解谱(37.5%,6/16),可裂解多种血清型大肠杆菌;最佳感染复数是0.1;具有较强的繁殖活性,潜伏期约为30 min,爆发量约为95PFU/cell;在40℃~50℃、pH 5~10之间噬菌体的效价较为稳定,对紫外线不耐受;基因组测序组装结果显示:PLYYY01基因组长度为57025 bp,(G+C)mol%含量为44.74%,共编码71个蛋白,未发现抗生素耐药基因及毒力因子基因。  相似文献   

8.
【目的】拟分离1株对大肠杆菌具有良好抑菌效果且适应性广泛的噬菌体,为开发新的大肠杆菌防治方法提供基础。【方法】以大肠杆菌H11为宿主菌,采用双层琼脂平板法从污水样品中分离纯化1株大肠杆菌噬菌体,电镜下观察噬菌体的形态,测定噬菌体的效价、宿主谱、最佳感染复数、一步生长曲线、pH稳定性与温度敏感性,对噬菌体的全基因组进行分析和序列比对,并探究该噬菌体在液体培养基中对H11的抑制效果。【结果】成功分离到1株裂解性大肠杆菌噬菌体,命名为EP_H11(登录号:OP688485)。双层琼脂平板法结果显示,该噬菌体能够形成大小一致、中间有透明亮点、周围有光晕的噬斑,噬菌体裂解效价可达109~1010 PFU/mL。电镜观察显示噬菌体EP_H11头部呈规则的多面体状,直径约85 nm,尾部直径约23 nm,长度约109 nm。以大肠杆菌H11为宿主菌测得EP_H11的最佳感染复数是0.1,一步生长曲线结果显示其潜伏期为40 min,爆发期为80 min,裂解量为200 PFU/cell。该噬菌体在pH 4.0~10.0的环境中至少可以存活2 h以上,并能够在...  相似文献   

9.
为了解噬菌体RAP44对鸭疫里默氏菌的侵染及裂解过程,本研究应用透射电镜进行观察。结果显示,噬菌体RAP44头部外廓呈六边形,平均直径约60nm,尾长约210nm,尾宽约8nm。RAP44与宿主菌混合后,5min内即能通过尾丝吸附于鸭疫里默氏菌的中部,开始感染过程;40min后细菌裂解,释放子代噬菌体。本研究为噬菌体作为抗生素替代品治疗鸭疫里默氏菌感染的研究提供了形态学依据。  相似文献   

10.
旨在为研制沙门菌特异性抗菌制剂提供试验数据和参考,以禽源沙门菌为宿主菌,从鸡场污水中分离纯化沙门菌噬菌体,并对其生物学特性和基因组学进行研究。结果显示:分离到1株沙门菌烈性噬菌体S5,透射电镜下该噬菌体头部直径约50 nm,尾部长约200 nm,头部呈正二十面体对称,属有尾噬菌体目、长尾噬菌体科;该噬菌体对20株沙门菌的裂解率为75%,效价为8×109PFU·mL-1;在30~60℃作用30 min和1 h的效价基本不变;pH 4.0~11.0对噬菌体活性影响不明显;最佳感染复数为0.000 1,一步生长曲线显示其感染宿主菌后5 min内(潜伏期)效价基本保持不变,暴发期约为95 min,平均裂解量为50 PFU·cell-1。噬菌体S5是双股DNA病毒,基因组全长为72 519 bp,GC含量39.45%,含有9种tRNA,预测到109个ORF,未发现任何编码细菌毒力因子等有害产物的基因。沙门菌烈性噬菌体S5,裂解效果较好、对环境适应性强、遗传背景清楚,以期成为防控沙门菌的抗生素替代产品。  相似文献   

11.
The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8 years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine®, Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive > 85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.  相似文献   

12.
Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm2, while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.  相似文献   

13.
实验利用透射电镜技术和扫描电镜技术对冷冻/解冻后的塔里木马鹿精子的超微结构进行了观察。结果表明:电镜下观察,塔里木马鹿精子头部呈扁平的卵圆形,长约5.95μm。颈部很短且不明显,长约0.45μm,宽为0.62μm。尾部中段横切面近圆形,直径约为0.58μm,轴丝为9×2+2型;线粒体鞘螺旋段为64~70转。尾部末段仅见质膜包围,9束微管和1对中央微管,形成9+2微管结构。冷冻/解冻后塔里木马鹿部分精子结构发生了变化,一些精子肿胀、质膜皱褶、扭曲,部分质膜损坏或溃散;顶体轻度肿胀,顶体外膜凸凹不平,形成突起。冷冻对头部破坏程度较小。  相似文献   

14.
《African Zoology》2013,48(2):192-201
The extent to which sexual dimorphism in snout–vent length, tail length and head shape (length, width and height) manifests itself in the legless fossorial skink subfamily Acontiinae was investigated in representatives of all four of its genera, Acontias, Microacontias, Acontophiops and Typhlosaurus. Where data were available fecundity selection and diet partitioning (ecological causation) were tested as possible proximate causes for dimorphisms found. The possibility of a relationship between head shape and microhabitat density was also investigated. The data showed that significant sexual dimorphism in snout–vent length and head shape was present in only a minority of taxa and that it was absent with respect to tail length. There are indications of a relationship between head shape and microhabitat density, and head shape seems to become increasingly more pointed as SVL increases. A serious impediment to placing the findings of this study into a firm evolutionary context is the general lack of information about the ecology, social organization and behaviour of the taxa.  相似文献   

15.
On assessment for use in an AI stud, a 12‐month‐old bull was found to produce low volume ejaculates with 41% of the sperm having morphological abnormalities. No left epididymal tail was palpable and the head of the epididymis on the left was twice the size compared with the right. Ultrasound examination showed the left testis to contain a large central area of decreased echogenicity, which could be followed proximally to a 15‐mm echolucent lesion at the site of the epididymal head. Postmortem examination revealed a 15‐mm diameter cyst in the region of the left epididymal head, and absence of the body and tail of the epididymis. The mediastinum testis of the left testis was dilated, corresponding to the area of decreased echogenicity observed on ultrasonography. No left seminal vesicle was present and the ampulla was significantly smaller than the same structure on the right. Histological examination revealed incomplete or absent spermatogenesis involving the majority of seminiferous tubules in the left testis, and a small proportion of those of the right testis. The cystic structure at the site of the left epididymal head was lined by irregular, sometimes attenuated, epithelium and contained sparse spermatozoa. This case demonstrates the adverse impact, which segmental aplasia of the mesonephric duct had on the testicular and epididymal function of a bull, and highlights the importance of careful clinical assessment in its diagnosis.  相似文献   

16.
基于细菌rRNA区域序列的PCR技术,通过设计特异性引物从奶样中快速、准确地鉴定奶牛乳房炎主要致病菌金黄色葡萄球菌。采用金黄色葡萄球菌与挤奶前从奶牛乳房上洗刷下来的污水经过3次富集的方法,通过双层琼脂平板法,分离纯化得到了强裂解性的奶牛乳房炎主要致病菌金黄色葡萄球菌的噬菌体。本试验分离的是可收缩尾的噬菌体,其头部为二十面体,约50 nm×62.5 nm,尾部长约187.5 nm,尾宽约37.5 nm,有尾鞘、基板等结构。纯化后在双层琼脂平板上形成的噬菌斑大而整齐,边缘光滑,透明,直径约2 mm,具有明显的强裂解性噬菌体特性,通过测定,该噬菌体的最佳感染复数为0.01,效价大于109 PFU/mL。对噬菌体的不同保存方法进行比较,结果表明,将噬菌体置于-80 ℃加入7% DMSO冷藏是较好的保存方法。  相似文献   

17.
Objective: To describe a technique used for reduction and internal fixation of sacral fractures in cattle. Study Design: Case series. Animals: Three heifers aged 19–32 months with deformation of the tail head and tail paralysis attributable to a closed fracture of S5 or S4 and S5. Methods: Fractures were reduced and repaired under general anesthesia. A narrow, 6–10‐hole, locking compression plate (LCP) was placed on the spinous processes of S5 and the first coccygeal (Co1) using 5.0 mm locking head screws. Follow‐up inquiries were made by telephone (3/3), and cows were examined clinically and radiographically 2 (1 cow) to 9 (1cow) months postoperatively. Results: No complications were encountered. The range of movement of the tail improved so that soiling of the tail during defecation was no longer a problem. In all heifers, the normal top line was restored and there was no narrowing of the pelvic canal. Unassisted calving occurred in all heifers, and follow‐up radiographs for 2 cows showed complete fracture healing. Conclusion: LCP‐osteosynthesis of the spinous processes provided sufficient stability for internal fixation of sacral fractures. Marked improvement in tail function occurred postoperatively without decompression by laminectomy.  相似文献   

18.
Ma YL  Lu CP 《Veterinary microbiology》2008,132(3-4):340-347
Streptococcus suis (S. suis) type 2 infection is considered to be a major problem worldwide in the swine industry. Studying phages of S. suis type 2 would be crucial for understanding the ecology and evolution of the Gram-positive bacteria. However, at the present, very little is known about them. An S. suis type 2 bacteriophage, named SMP, was isolated from nasal swabs of healthy Bama minipigs and was characterized at the microbiological and molecular levels. Phage SMP had an isometric head of 50 nm, a noncontractile tail of approximately 135 nm, and a linear double-stranded DNA genome. The host range of phage SMP was limited to 2 of 24 S. suis type 2 strains tested. The genome of phage SMP contained 36,216 bp with an average G+C content of 41.6%.  相似文献   

19.
1株肠炎沙门氏菌噬菌体的污水分离及生物学特性研究   总被引:2,自引:2,他引:0  
Using double-layer plate method, a phage against Salmonella enteritidis was isolated from sewage water, and its biological characteristics were studied. The plaques of isolated phage were transparent with clear boundary, and were about 2 mm in diameter at 12 h. The morphological properties of phage were examined by electron microscope, and showed that the phage had an isometric polyhedral no-enveloped head with diameter of about 54 nm, and a long noncontractile tail with size of about 110 nm. The test of optimal multiplicity of infection showed when MOI equaled 2, the number of phage offspring was higher. One-step growth kinetics was determined according to the results of one-step growth experiment, which showed that the latent period was about 10 min, the rise period was about 100 min, and the average bust size was about 61. The phage against S.enteritidis was successfully isolated from the sewage water in this study, which would provide materials for the further biological control researches of S.enteritidis using corresponding phage.  相似文献   

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