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1.
为了解山东省滕州市羊隐孢子虫感染情况和种类,从该市2个绵羊场、2个山羊场采集222份羊粪便样品,提取所有样品的基因组DNA,采用巢式PCR扩增隐孢子虫actin基因,对阳性样品进行序列测定和分析。结果显示,滕州市羊隐孢子虫总的感染率为6.76%,其中绵羊7.84%,山羊5.83%;不同月龄的羊感染情况不同,6~12月龄绵羊感染率最高(20.69%),而12月龄以上的山羊感染率最高(10.00%);共发现3种隐孢子虫感染,其中微小隐孢子虫(Cryptosporidium parvum,C.parvum)检出率最高为53.33%,其次是肖氏隐孢子虫(C.xiaoi)为40.00%,仅发现1例(6.67%)泛在隐孢子虫(C.ubiquitum)感染;序列比对结果显示,本试验所获得基因序列与Gen Bank中的C.parvum、C.xiaoi、C.ubiquitum actin基因序列同源性达到100%;系统进化分析显示,滕州市的羊样品中鉴定的actin序列,与Gen Bank中的相应虫种序列在同一分支上。研究结果为深入了解滕州市羊隐孢子虫流行情况及制定有效的防控措施提供了依据。  相似文献   

2.
为了解宁夏回族自治区吴忠市羊隐孢子虫病流行情况,从吴忠市的2个规模羊场采集480份粪便样品。提取基因组DNA,用套式PCR扩增隐孢子虫SSUrRNA基因,阳性样品进行测序分析。结果显示,吴忠市羊隐孢子虫感染率为28.3%,其中盐池县绵羊感染率为33.8%,同心县山羊感染率为22.9%;共发现3种隐孢子虫感染,其中肖氏隐孢子虫(Cryptosporidium xiaoi)阳性率最高(62.5%),其次是泛在隐孢子虫(C.ubiquitum)(32.4%),微小隐孢子虫(C.parvum)感染率最低(5.2%);从季节上看,春季感染率最高(49.2%),夏季最低(6.7%);从羊的年龄上看,未断奶羔羊感染率最高(34.4%),而成年羊感染率最低(19.4%);序列比对结果显示,所有阳性样品均在Gen Bank找到100%同源的序列。本研究首次对宁夏羊感染的隐孢子虫进行了分子鉴定,为深入了解宁夏吴忠市羊隐孢子虫病流行情况、制定有效的防控措施提供了依据。  相似文献   

3.
《中国兽医学报》2017,(4):686-691
为了阐明陕北黑山羊隐孢子虫(Cryptosporidium)的感染情况、种类及其遗传进化关系,本研究利用形态学和分子生物学方法,分析了107份不同年龄段黑山羊粪便样品中的隐孢子虫的感染状况、种类以及遗传进化关系。结果发现,黑山羊存在隐孢子虫感染,感染率为14%(15/107)。序列比对分析表明,7份阳性样品为牛隐孢子虫(C.bovis)感染,6份为肖氏隐孢子虫(C.xiaoi)感染,2份为C.bovis和C.xiaoi混合感染。系统进化树显示9个分离株与C.bovis位于同一分支,而与C.xiao位于同1个分支的有8个分离株。结果为陕北黑山羊乃至其他山羊的隐孢子虫病分子流行病学研究及防控提供了理论基础。  相似文献   

4.
为探讨四川部分地区不同养殖模式下成年山羊隐孢子虫的感染差异以及感染虫种基因型,分别采集了规模化养殖模式和散养模式各6个养殖场共342份新鲜粪便,采用改良饱和蔗糖溶液漂浮法处理后提取粪便DNA,经套式PCR扩增18S rRNA基因,产物测序后进行遗传进化分析。结果:采样地区山羊隐孢子虫总感染率为4.7%,12个养殖场中4个养殖场(名山、纳溪、邻水和双流)存在隐孢子虫感染,感染率2.5%～19.2%,且4个隐孢子虫感染阳性场均为散养模式。序列分析表明,感染虫种为肖氏隐孢子虫(Cryptosporidium xiaoi)和猪隐孢子虫(C.suis),并以肖氏隐孢子虫为主(68.7%)。本试验初步表明,规模化养殖模式和散养模式下山羊隐孢子虫感染存在差异(P0.01);首次在成年山羊中检出猪隐孢子虫虫种,序列分析表明所获得的猪隐孢子虫序列与人源猪隐孢子虫序列完全一致,提示此次分离的羊源猪隐孢子虫为人兽共患隐孢子虫种。  相似文献   

5.
河南省山羊隐孢子虫流行病学调查   总被引：1，自引：1，他引：0  
对河南省4个山羊场进行了隐孢子虫病流行病学调查.采集山羊粪样1 017份,以饱和蔗糖漂浮法和改良抗酸染色法检查,结果共检出隐孢子虫卵囊阳性山羊粪样28份,平均感染率为2.75%;有2个山羊场检出隐孢子虫卵囊,场感染率为50%;中牟某羊场阳性率最高,达4.95%;在各年龄段中,2～4月龄山羊隐孢子虫感染率最高,达5.43%;山羊隐孢子虫感染可能与季节有关;所检出的隐孢子虫卵囊经形态学观察,初步鉴定为微小隐孢子虫(Cryptosporidium parvum).  相似文献   

6.
为了解宁夏地区犊牛隐孢子虫的感染情况,从3个市8个规模化奶牛场采集272份犊牛粪便样品,利用巢氏PCR技术对18S rRNA基因位点进行检测和分析。结果显示,隐孢子虫的总感染率为12.88%,银川市隐孢子虫感染率为16.89%,吴忠市隐孢子虫感染率为6.94%,石嘴山市隐孢子虫感染率为9.62%,三者差异不显著(P0.05),断奶前、后感染率分别为14.14%,12.14%。序列处理分析发现3种隐孢子虫,分别为牛隐孢子虫(Cryptosporidium bovis,C.bovis)、瑞氏隐孢子虫(Cryptosporidium ryanae,C.ryanae)和微小隐孢子虫(Cryptosporidium parvum,C.parvum),其中C.bovis分离率最高(54.29%)。研究表明这些地区犊牛隐孢子虫感染比较普遍,且感染的虫种有3种,其为深入了解宁夏部分地区隐孢子虫的分子流行情况提供了参考数据。  相似文献   

7.
为了解我国西北部分地区藏香猪隐孢子虫的感染和种类分布,从陕西省和青海省采集了共450份藏香猪新鲜粪便样品,基于隐孢子虫18S rRNA基因的分子生物学方法对粪便样品中隐孢子虫感染情况及其种类进行了研究。结果显示,藏香猪隐孢子虫总感染率为14.2%,青海省藏香猪的感染率(23.9%)显著高于陕西省(3.7%)(P0.01);不同年龄段藏香猪隐孢子虫感染率差异显著(P0.01),其中断奶后仔猪感染率(42.9%)最高,而哺乳仔猪和成年猪均未检测到隐孢子虫的感染;序列分析发现,藏香猪的隐孢子虫均为Cryptosporidium scrofarum。研究结果为藏香猪隐孢子虫病的防控提供了基础数据。  相似文献   

8.
《畜牧与兽医》2014,(9):94-95
为了解安徽省山羊隐孢子虫感染情况,为制定人畜共患隐孢子虫病的防控措施提供理论依据,在安徽省境内选泗县、涡阳县和霍邱县作为调查点,分别从各调查点山羊养殖场随机采集50、50、30份新鲜山羊粪样,用改良抗酸染色方法进行检测。共检获2种隐孢子虫,即微小隐孢子虫(Cryptosporidium parvum)和安氏隐孢子虫(C.andersoni);隐孢子虫阳性粪样均源自泗县,其感染率为36.00%(18/50),在涡阳县、霍邱县山羊粪样中未检出隐孢子虫。结论表明:羊感染隐孢子虫与饲养管理、自身免疫力等相关。  相似文献   

9.
为初步了解水貂隐孢子虫病的流行情况,作者于2005年11月份用饱和蔗糖溶液漂浮法检查了河北省肃宁县某水貂养殖场的469份粪便样品。结果,8份粪便样品为隐孢子虫阳性,总感染率为1.71%(8/469)。其中,白貂感染率为2.15%(5/233)、灰貂感染率为2.08%(1/48)、黑貂感染率为1.06%(2/188)。所查的8份隐孢子虫阳性样品均来自5~6月龄的水貂,表明幼龄水貂容易感染隐孢子虫病而老龄水貂不易感染。另外,8份阳性样品多数来自雄性水貂,显示水貂的隐孢子虫感染可能存在性别的差异性。根据卵囊形态和大小将水貂隐孢子虫初步鉴定为小球隐孢子虫(Cryptosporidium parvum)。同时,利用所收集的隐孢子虫卵囊进行了小白鼠感染试验,结果表明水貂源隐孢子虫不感染免疫抑制状态下的昆明系小白鼠。  相似文献   

10.
陕西部分地区秦川牛断奶后犊牛隐孢子虫感染状况   总被引：1，自引：0，他引：1  
为明确陕西部分地区秦川牛断奶后犊牛隐孢子虫感染率及其种类分布,本试验从杨凌和宝鸡3个秦川牛场采集了113份3~6月龄的犊牛粪便样品,基于隐孢子虫18SrRNA基因位点对其进行PCR检测和序列分析。结果显示,隐孢子虫总感染率为16.8%(19/113),各养殖场的隐孢子虫感染率差异不显著(P0.05);种类鉴定发现了3种隐孢子虫,分别为安氏隐孢子虫(Cryptosporidium andersoni)、瑞氏隐孢子虫(Cryptosporidium ryanae)和牛隐孢子虫(Cryptosporidium bovis),其中安氏隐孢子虫为优势虫种,广泛分布于各个养殖场。这些研究结果表明这些地区的秦川牛断奶后犊牛隐孢子虫感染比较普遍,且种类较为复杂。  相似文献   

11.
Faecal specimens were taken from 205 sheep and goats housed in five different localities in the west-central part of Poland. All faecal specimens were examined for Cryptosporidium by using microscopy screening of smears stained by modified Ziehl-Neelsen technique and commercial enzyme immunoassay. PCR technique using genus specific primers was additionally applied in the surveys of 10 faecal specimens collected from lambs. C. parvum infection was identified in 16 of 159 sheep (10.1%). Lambs were more often infected than adult sheep, and the intensity of infection was higher in lambs than in sheep, as a rule. Both lambs and sheep examined in the study were asymptomatically infected with Cryptosporidium. Both microscopy and enzyme immunoassay methods gave one false negative result. The examination of 10 faecal samples revealed 100% agreement among the results obtained by microscopic, immunologic and molecular methods. None of the goats raised on three farms were infected with Cryptosporidium.  相似文献   

12.
Diarrheic fecal samples from 258 pre-weaned calves (1-30 day-old) from 9 dairy farms located in Banat region, Romania, were microscopically examined for the presence of Cryptosporidium oocysts. Overall, 65 (25%) samples were found positive. A higher percent of infection was recorded in calves aged between 8 and 14 days compared with other age categories (1-7, 8-14, 15-21 and 22-30 days; p<0.05). Genetic characterization was carried out on all Cryptosporidium-positive samples. After DNA extraction, Cryptosporidium species were determined by a nested PCR of the small subunit rRNA gene (18S) followed by RFLP analysis with SspI, VspI and MboII restriction enzymes. The restriction patterns showed that animals were infected with Cryptosporidium parvum. Subsequently, subtyping of 13 C. parvum isolates, based on sequence analysis of the 60 kDa glycoprotein (GP60) gene, showed 2 subtypes (IIaA15G2R1 and IIaA16G1R1) belonging to the subtype family IIa. This is the first molecular study of bovine Cryptosporidium infection in Romania.  相似文献   

13.
Data of the prevalence, age-related and housing-dependence of naturally acquired cryptosporidiosis on 11 dairy and 11 beef farms in South Bohemia (Czech Republic) were collected. The farms were visited over four consecutive years (from 2002 to 2005). The prevalence of Cryptosporidium in pre-weaned (animals until second month of age) and post-weaned (animals from the third month of age) calves was determined. A total of 7001 faecal samples were collected, concentrated by Sheather's floatation method and stained by aniline-carbol-methyl violet. All samples were examined by light microscopy. Cryptosporidium parvum and C. andersoni oocysts were differentiated on morphological criteria. Of the 7021 specimens, 1814 (25.8%) were positive for Cryptosporidium oocysts; 561 samples (8%) for C. parvum and 1253 (17.8%) for C. andersoni. Pre-weaned dairy calves had higher infection levels of C. parvum than pre-weaned beef calves. The prevalence of C. parvum ranged from 1.4 to 56.5% on dairy farms. Only three cases of C. parvum oocysts shedding in pre-weaned calves on beef farms were found. Only one case of C. andersoni infection in pre-weaned calves was detected and no infections of C. parvum in post-weaned calves were found. The prevalence of C. andersoni reached 35.5% on dairy farms and 61.7% on beef farms. Calves that were on pasture all year long, had a lower probability of C. andersoni infection than those calves kept in a cowshed during the winter season.  相似文献   

14.
The prevalence of Cryptosporidium and Giardia was studied on 10 intensively reared sheep and goat farms in the province of East Flanders, Belgium. Random faecal samples were collected and examined using the Merifluor((R)) immunofluorescence assay. Cryptosporidium positive samples were withheld for molecular identification using primers targeting the 18S rDNA, 70 kDa heat shock protein and 60 kDa glycoprotein gene. For the molecular identification of Giardia the beta-giardin gene and a recently developed assemblage specific PCR based on the triose phosphate isomerase gene were used. The prevalence of Cryptosporidium in lambs was 13.1% (18/137), on 4 out of 10 farms. In goat kids the Cryptosporidium prevalence was 9.5% (14/148), on 6 out of 10 farms. The molecular characterisation of Cryptosporidium positive isolates indicated that in lambs (n=10) the cervine genotype was predominant, whereas in the goat kids (n=11) only C. parvum was identified, with subgenotypes IIaA15G2R1 and IIdA22G1. The Giardia prevalence was 25.5% (35/137) in lambs with all 10 farms being positive, and 35.8% (53/148) in goat kids with 8 out of the 10 farms being positive. Both in the goat kids and in the lambs the host specific assemblage E was most commonly identified. However, the zoonotic assemblage A was identified in 6 out of 28 goat kids and in 2 out of 8 lambs, based on the beta-giardin sequence alignment. Using the assemblage specific PCR, mixed assemblage A and E infections were additionally identified in 2 lambs and in 5 goat kids. The results of the present study indicate that both Cryptosporidium and Giardia are common parasites on intensively reared sheep and goat farms in the province of East Flanders, Belgium, and that they are a potential source for zoonotic infections.  相似文献   

15.
为了解规模化舍饲湖羊消化道寄生虫感染情况,本研究应用饱和蔗糖溶液漂浮法、饱和盐水漂浮法、卢戈氏碘液染色法、离心沉淀法和麦克马斯特氏计数法等对采自河南部分地区规模化全舍饲湖羊养殖场共计553份粪便样品进行了调查。调查结果显示：寄生虫总感染率高达97.47%,75.23%的样品混合感染,样品混合感染的寄生虫种类最多为5种;共查到球虫、隐孢子虫、贾第虫、阿米巴、鞭虫、圆线虫和绦虫7种寄生虫,感染率分别为90.42%、0.90%、4.88%、65.64%、12.48%、42.13%和4.88%;感染强度最大的为球虫,每克粪便的卵囊数（OPG）最高达652 000,其次为圆线虫,每克粪便的虫卵数（EPG）最高为7 000;湖羊消化道寄生虫感染无明显的年龄、性别差异（P>0.05）;季节流行动态显示,春、夏、秋三季的寄生虫感染率与冬季相比有较大差异。以上结果说明,湖羊消化道寄生虫感染较为普遍,应采取有效的综合防控措施,以保障羊群的健康发展。  相似文献   

16.
A total of 364 fecal specimens from randomly selected pre-weaned calves, aged up to 4 months, from 5 different farms in the south of Western Australia and 1 farm from New South Wales were screened for the presence of Cryptosporidium and Giardia using PCR. There were substantial differences in prevalence between the farms and the overall prevalence was 22.3% (81/364) and 26.9% (98/364) respectively for Cryptosporidium and Giardia. For Cryptosporidium, 70 positives were identified at the 18S locus. At a unique diagnostic locus, an additional 12 C. parvum positives were identified. Sequence analysis at the 18S ribosomal RNA locus was successful for 59 of the 70 positive isolates; of these 14 were C. parvum, 28 were C. bovis, 15 were C. ryanae, 1 was pig genotype II and 1 was a mixed C. ryanae/C. parvum infection. Sub-typing analysis at the glycoprotein 60 (gp60) locus for 24 C. parvum isolates identified all as IIa; 17 were A17G2R1, 1 was A18G3R1 and 6 were A20G3R1. For Giardia, 75 positives were identified at the 18S locus and an additional 23 positives were identified at the gdh locus. The majority of the isolates sequenced were assemblage E, however assemblage A and B and mixed A and E and A, B and E infections as well as the quenda genotype were identified. The findings of the present study indicate that pre-weaned calves are not an important source of zoonotic Giardia species in Australia but may be an important source of zoonotic Cryptosporidium.  相似文献   

17.
An epidemiologic study was carried out to investigate the prevalence of and to identify factors associated with the risk of Cryptosporidium infection in sheep in Zaragoza (northeastern Spain). Faecal samples from 583 lambs aged from 1 day to 3 months and 205 ewes older than 1 year were collected at 89 farms in the two regions of the province of Zaragoza with the highest sheep population (Zaragoza and Ejea de los Caballeros). In every sheep farm, data of the factors potentially associated with the likelihood of C. parvum infection were analysed: geographical location, season, size of herd, number of lambs in the herd at sampling time, lambing period, cleaning of lambing area and presence of diarrhoeic lambs in the farm. C. parvum oocysts were identified by using the Ziehl-Neelsen technique in 344 lambs (59%) from 75 farms (84.4%). Infected lambs ranged from less than 7 days to 90 days of age, although the percentage of animals shedding oocysts peaked at 8-14 days of age (76.2%). Statistical analysis showed that infection rates were significantly higher in lambs aged between 1 and 21 days (66.4%) than in those aged between 22 and 90 days (23%) (P<0.0001, chi(2)). Analysis of correlation between excretion of oocysts and diarrhoea revealed a relationship in all age groups and the probability of presenting diarrhoea was significantly higher for lambs shedding oocysts (86.3%) than for those which did not excrete the parasite (32.2%) (P<0.0001, chi(2)). Similarly, cryptosporidial infection rates were significantly higher in diarrhoeic (79.4%) than in non-diarrhoeic lambs (22.4%). Furthermore, infection intensity was correlated with the presence of clinical symptoms. Presence of diarrhoeic lambs in the farm was the only factor significantly associated with an increased risk of infection since the percentage of herds testing positive was significantly higher in farms with diarrhoeic lambs (91.3%) than in those without cases of neonatal diarrhoea (12.5%) (P<0.0001, chi(2)). Factors associated with a decreased risk of C. parvum infection in lambs included low numbers of lambs in the farm and cleaning of the lambing area. Additionally, lambs 8-14 days of age were less likely to be infected at the first lambing period and in spring/autumn. Cryptosporidial infection was also detected in 16 ewes (7.8%) which excreted few oocysts and without diarrhoea.  相似文献   

18.
The prevalence of Cryptosporidium species in 1-2-year-old heifers was determined for 571 animals on 14 dairy farms in seven states on the East Coast of the United States. A fecal specimen collected directly from each heifer was processed to concentrate oocysts that were then examined by polymerase chain reaction (PCR). For every PCR-positive specimen the 18S rRNA gene of Cryptosporidium was sequenced. Cryptosporidium was identified by PCR from heifers on 13 of 14 farms. On all except four farms groups of heifers were housed in a barn or in large covered pens. Others were pastured. From many of the same farms an earlier study reported that 41% of 393 pre-weaned calves and 26.2% of 447 post-weaned calves were infected. In the present study, 11.9% of 571 heifers were infected with Cryptosporidium, 0.7% with Cryptosporidium parvum, the zoonotic species. Of 68 PCR-positive specimens characterized by gene sequencing 1, 4, 10, 24, and 29 calves were infected with Cryptosporidium suis, Cryptosporidium parvum, Cryptosporidium deer-like genotype, Cryptosporidium bovis, and Cryptosporidium andersoni, respectively. These findings demonstrate a lower prevalence of infection in 1-2-year-old dairy cattle than in younger cattle as well as a change in the diversity of species present. Consequently, the risk of humans acquiring infection with C. parvum from exposure to feces from yearling and older cattle appears much lower than from exposure to pre-weaned calves.  相似文献   

19.
During the kidding season between January and April 2003, 10 farms were selected and divided into two groups of five. The farms in group A had had serious diarrhoeal illness and losses in neonatal kids the previous year, and there were Cryptosporidium parvum infections in kids associated with diarrhoea during the survey. On the farms in group B, there was no history of diarrhoeal disease the previous year and neither C parvum oocysts nor diarrhoea were detected in neonatal kids during the survey. Faecal samples were collected once from 10 adult goats aged between one and seven years on each farm. To assess more accurately the pattern of output of oocysts of C parvum and cysts of Giardia duodenalis by periparturient adult goats, one farm was selected from each group, faecal samples were collected weekly before and after kidding from 12 goats on the farm in group A and from 10 goats on the farm in group B. There was no significant difference in the prevalence of G duodenalis cysts between the group A farms (14 per cent) and the group B farms (12 per cent), and the numbers of cysts excreted ranged from 143 to 400 cysts per gram of faeces (cpg) on the group A farms and 72 to 334 cpg on the group B farms. There was a significant difference (P=0.03) in the prevalence of C parvum oocysts at the group level between the group A farms (20 per cent) and the group B farms (6 per cent). All the adult goats excreted cysts and oocysts at some date around the kidding period; the number of animals excreting cysts of G duodenalis or oocysts of C parvum increased when they gave birth, and seven to 10 times more cysts and oocysts were shed in the three weeks around kidding than in the period more than three weeks from kidding (P<0.001).  相似文献   

20.
Three LAMP (loop-mediated isothermal DNA amplification) assays were applied to detect Cryptosporidium species DNA in a total number of 270 fecal samples originating from cattle, sheep and horses in South Africa. DNA was extracted from 0.5 g of fecal material. Results of LAMP detection were compared to those obtained by nested PCR targeting the Cryptosporidium 18 small subunit rRNA (18S) gene. All samples were negative by nested PCR, while up to one-third of samples were positive by LAMP assays. The SAM-1 LAMP assay, shown to detect C. parvum, C. hominis and C. meleagridis, amplified Cryptosporidium DNA in 36 of 107 cattle (33.64%), in 26 of 85 sheep (30.5%) and in 17 of 78 horses (21.79%). The HSP LAMP specific to C. muris and C. andersoni, amplified Cryptosporidium DNA in one cow (0.9%), five sheep (5.8%) and seven horses (8.9%). The gp60 LAMP assay, shown to detect C. parvum produced no amplified Cryptosporidium DNA, likely due to low sample DNA concentrations. The specificity of LAMP assays was confirmed by sequencing of the LAMP products generated in positive samples. Sequence products from the three LAMP assays showed high identity to the target gene sequences confirming the specificity of LAMP. In this study, the LAMP procedure was clearly superior to nested PCR in the detection of Cryptosporidium species DNA. Use of LAMP is proposed as an efficient and effective tool for epidemiologic survey studies including screening of healthy animals in which Cryptosporidium oocyst shedding is characteristically low and likely below the detection limit of PCR in conventional sample concentrates.  相似文献   

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