copro-PCR

Copro - PCR
  • 文章类型: Journal Article
    在过去的30年里,法国的灰狼数量已经恢复,最初是来自意大利的狼穿过阿尔卑斯山。人口受到严密监控,但是关于他们的蠕虫动物的信息很少,其中包括具有公共卫生重要性的寄生虫:多房棘球蚴和细粒棘球蚴。利用收集的911个粪便样本对法国狼种群进行非侵入性遗传监测,连同15只死狼的肠子,在法国狼中评估了棘球蚴物种以及其他蠕虫物种的存在。将扩增大量寄生虫的copro-PCR方法用于粪便样品,而使用SCT分析肠。粪便中出现严格的细粒E.granulosussericto(2.4%)和多房性E.和其他寄生物种,与其他欧洲狼种群相似,包括棘球蚴(7.2%),牛带虫(2.4%),海螺(2.4%),马尾藻(1.9%),牛带虫(0.3%),多头肌带虫(0.1%),和弓形虫(0.1%)。在肠道中也发现了三种最丰富的物种。S.granulosussensustricto的感染与狼群区域和绵羊繁殖牧区的重叠一致。然而,狼似乎在严格的E.granulosussensu的生命周期中没有发挥重要作用。在法国东南部,这种机会性粪便样本的可用性意味着,在近年来观察到的向南扩张的背景下,它们可以用作监视多房菌的哨兵。
    Over the past 30 years, the gray wolf population has recovered in France, initially to wolves from Italy passing through the Alps. The population is carefully monitored, but little information is available on their helminth fauna, which includes parasites of public health importance: Echinococcus multilocularis and Echinococcus granulosus sensu lato. Capitalizing on the availability of 911 fecal samples collected for the noninvasive genetic monitoring of French wolf populations, along with the intestines from 15 dead wolves, the presence of Echinococcus species among others helminth species was evaluated in French wolves. A copro-PCR approach amplifying a large spectrum of parasites was used for fecal samples while intestines were analyzed using SCT. The fecal occurrences of E. granulosus sensu stricto (2.4%) and E. multilocularis (0.3%), and indeedother parasitic species, are similar to those of other European wolf populations including Taenia hydatigena (7.2%), Taenia krabbei (2.4%), Uncinaria stenocephala (2.4%), Mesocestoides litteratus (1.9%), Taenia ovis (0.3%), Taenia multiceps (0.1%), and Toxascaris leonina (0.1%). The three most abundant species were also found in the intestines. Infections by E. granulosus sensu stricto are in accordance with the overlap of wolf pack areas and sheep breeding pastoral units. However, the wolf does not appear to play a significant role in the lifecycle of E. granulosus sensu stricto. The availability of this opportunistic fecal sampling of wolves in southeastern France means that they can be used as sentinels for the surveillance of E. multilocularis in the context of its southward expansion observed in recent years.
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  • 文章类型: Journal Article
    在福克兰群岛,控制细粒棘球蚴引起的囊性棘球蚴病(CE)的尝试已经进行了50多年。自1980年代以来,没有人类病例记录,但有必要确定这种寄生虫是否已从家畜中完全消除。在2018/2019年进行了一项研究,以使用共抗原和共聚合酶链反应(PCR)分析来鉴定感染细粒大肠杆菌的狗。此外,对每年的屠宰数据进行分析,以确定细粒大肠杆菌和2种其他大黄鱼寄生虫的感染水平.结果显示589只狗中的4只(0.7%)通过共同抗原分析检测为阳性。2010年、2012年和2014年进行的类似调查的结果显示17(3%),0和6只(1%)共同抗原阳性的狗,分别,2010年通过PCR确认了8只狗。每年的屠宰场数据显示,从2006年到2020年,36只绵羊被鉴定为E.granulosus(平均0.0055%),14186只带棘球蚴的绵羊(平均2.2%)和465只带棘球蚴的绵羊(平均0.072%)。T.hyhypa和T.ovis的患病率在某些年份显示出自发的上升,在羔羊中也可以检测到感染,这表明存在活的taeniid卵。对农场管理程序的观察表明,有时狗可以接触到感染性大黄鱼。总之,E.granulosus仍然存在于绵羊和狗中,但患病率较低。然而,水母的存在越来越多,表明控制措施在某些领域存在缺陷,并且有可能重新出现CE。
    Attempts to control cystic echinococcosis (CE) caused by Echinococcus granulosus in the Falkland Islands have been ongoing for over 50 years. No human cases have been recorded since the 1980s but there is a need to establish if the parasite has been completely eliminated from domestic animals. A study was carried out in 2018/2019 to identify dogs infected with E. granulosus using copro-antigen and copro-polymerase chain reaction (PCR) analysis. In addition, annual slaughter data were analysed to establish infection levels of E. granulosus and 2 other taeniid parasites. Results showed that 4 out of 589 dogs (0.7%) tested positive by copro-antigen analysis. Results from similar surveys carried out in 2010, 2012 and 2014 showed 17 (3%), 0 and 6 (1%) copro-antigen-positive dogs, respectively, with 8 dogs being confirmed by PCR in 2010. Annual abattoir data showed that from 2006 to 2020, 36 sheep were identified with E. granulosus (mean 0.0055%), 14 186 sheep with Taenia hydatigena (mean 2.2%) and 465 with Taenia ovis (mean 0.072%). Prevalences of T. hydatigena and T. ovis showed spontaneous rises in certain years where the infections could also be detected in lambs indicating that viable taeniid eggs were present. Observations of farm management procedures indicated that there were occasions when dogs could get access to infective taeniid material. In conclusion, E. granulosus is still present in sheep and dogs but at low prevalences. The increasing presence of T. hydatigena however, indicates that control measures are defective in some areas and there is potential for a re-emergence of CE.
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  • 文章类型: Journal Article
    Cystic Echinococcosis (CE) is endemic in humans and livestock in many pastoral communities in Kenya. The distribution of the disease is enhanced by several factors, including livestock trade, which has allowed for the spread of CE to non-endemic areas such as western Kenya. Dogs\' roaming behaviour, with consequent contamination of the environment with intestinal parasites, could then lead to parasite establishment. This study examined dogs\' infection levels with taeniid eggs and their potential role in contaminating the environment with intestinal parasites.
    We selected sixteen ruminant slaughterhouses in Busia and Bungoma Counties, and around each slaughterhouse we identified ten homesteads owning free-roaming dogs. We administered a questionnaire on dog management practices to the homestead owner and collected a faecal sample from the dog\'s rectum. In homesteads around 8 of the 16 slaughterhouses, we collared dogs with a GPS tracker to assess their movement patterns. The faecal samples were examined microscopically following zinc-chloride sieving-floatation technique for the presence of taeniid eggs and other canine intestinal parasites. Polymerase Chain Reaction - Restriction Fragment Length Polymorphism of NADH dehydrogenase subunit 1 gene and sequencing were used to confirm taeniid eggs identified during microscopy. Additionally, the Coproantigen-ELISA was used to detect the presence of taeniid antigen in a sub-set of the faecal samples.
    Helminths detected in the 155 dogs sampled included hookworms (n = 92; 59.4%), ascarids (n = 15; 9.7%), and taeniids (n = 1; 0.6%). Through Copro-PCR, 13 eggs extracted from the sample of the only taeniid infected dog were sequenced and identified as E. canadensis (G6/7) [n = 1], Taenia multiceps [n = 1], and Taenia serialis [n = 6]; the remaining were indeterminate. Of the 77 faecal samples tested for E. granulosus sensu lato (s. l.) with the Copro-ELISA, 64 (83.1%) were negative, 12 (15.6%) were positive, while 1 (1.3%) was suspicious. The dogs travelled a median of 13.5 km daily, and 28 dogs visited the slaughterhouses during the 5-day recording period.
    The results indicate a relatively high carriage of zoonotic parasites by free-roaming domestic dogs in western Kenya, which poses a risk to human and livestock populations. We report for the first time a domestic lifecycle of Echinococcus canadensis and Taenia multiceps in western Kenya, as well as a presumptive sylvatic cycle of coenurosis by T. serialis. We recommend an extensive and ongoing Copro-antigen survey of dog faeces, broader assessment of dog parasites with zoonotic potential, adherence to slaughterhouse management practices, and dog-ownership programmes to highlight the importance of deworming and restricted dog movements.
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  • 文章类型: Journal Article
    浣熊(Procyonlotor)和浣熊犬(Nyctereutesprocyonoides)被引入欧洲,在过去的几十年里,他们的人口增加并适应了整个欧洲的同人化环境。鉴于它们可能在欧洲进一步传播,入侵物种应该被视为蠕虫的潜在水库,包括tape虫。这项研究旨在通过基于copro-DNA分析鉴定c虫物种,来调查波兰引进的野生食肉动物中and虫的患病率和多样性。总共214个来自非本地侵袭性食肉动物的粪便样本,即,浣熊和浣熊狗,另外还有47个来自本地食肉动物的样本,即,欧洲badge(Melesmeles),分析了昆虫的存在。针对线粒体(mt)12SrRNA基因片段的粪便样品的PCR分析显示,103只浣熊中有19只(18.4%),111只浣熊狗中的13只(11.7%)和47只badge中的23只(48.9%)。序列分析表明,浣熊和浣熊犬中存在Mesocestoideslitteratus,而仅在来自浣熊犬的两个样本中鉴定出了线性中胚层。此外,在这项研究中,首次通过使用mt12SrRNA基因的片段进行分子表征,在所有被检查宿主物种的粪便样本中都检测到了这种昆虫物种的DNA。
    The raccoon (Procyon lotor) and the raccoon dog (Nyctereutes procyonoides) were introduced to Europe and, in the past decades, their populations have increased and adapted to synanthropic environments across Europe. In view of their possible further spread in Europe, the invasive species should be examined as potential reservoirs for helminths, including tapeworms. This study aims to investigate the prevalence and diversity of tapeworms in introduced wild carnivores in Poland by identifying cestode species based on copro-DNA analysis. A total of 214 individual fecal samples from non-native invasive carnivores, i.e., raccoons and raccoon dogs, and additionally 47 samples from native carnivores, i.e., European badgers (Meles meles), were analyzed for the presence of cestodes. PCR analysis of fecal samples targeting a fragment of mitochondrial (mt) 12S rRNA gene revealed the presence of cestode DNA in 19 of 103 (18.4%) raccoons, in 13 of 111 (11.7%) raccoon dogs and in 23 of 47 (48.9%) badgers. Sequence analysis demonstrated the presence of Mesocestoides litteratus in raccoons and raccoon dogs, while Mesocestoides lineatus was identified only in two samples derived from raccoon dogs. Moreover, in this study, Atriotaenia incisa was for the first time molecularly characterized by using fragments of mt 12S rRNA gene, and the DNA of this cestode species was detected in the fecal samples of all the examined host species.
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  • 文章类型: Journal Article
    BACKGROUND: The pastoral area of the eastern Tibetan Plateau is highly endemic for human echinococcosis. Domestic dogs are the main definitive host for the transmission of both Echinococcus granulosus (sensu lato) and E. multilocularis to humans. To control the infection risks, a national-level canine echinococcosis prevention and control programme has been implemented since 2015 in Shiqu County, Ganze Tibetan Autonomous Prefecture, Sichuan, China. The objective of this study was to evaluate its effect on Echinococcus spp. prevalence in dogs.
    METHODS: We surveyed 69 households with 84 owned dogs, for dog fecal samples and dog keeping information in the villages of Rizha and Eduoma. A total of 105 dog fecal samples (75 from owned dogs and 30 unknown dog fecal samples) were collected between 2015-2017 to determine Echinococcus spp. prevalence using copro-PCR. Eight variables based on household surveys were included into a logistic regression model for significant risk factors to canine echinococcosis prevalence in dogs.
    RESULTS: Between 2015-2017, the overall Echinococcus spp. copro-DNA prevalence decreased significantly in dogs from 51.2% (2015) to 20.0% (2017) in Rizha, and insignificantly from 11.5% (2016) to 4.3% (2017) in Eduoma. Echinococcus multilocularis was the most prevalent species continually copro-DNA detected during the entire study period, while E. granulosus was rare and not detected in 2017. Echinococcus shiquicus copro-DNA prevalence (a probable non-zoonotic wildlife species) was as high in dogs as that of E. multilocularis, although only detected in 2015 in Rizha. Unleashed dog feces were mainly collected in Rizha in 2015. Although 93.2% of owned dogs were leashed, and the monthly praziquantel dosing rate reached 97%, E. multilocularis infection could still be detected in 11.1% of owned dogs in 2017. Monthly deworming, leashing dogs 24 h per day, and the avoidance of dogs feeding on livestock viscera were significant measures to prevent canine echinococcosis infection in owned dogs.
    CONCLUSIONS: Carrying out a canine echinococcosis prevention and control programme can significantly decrease Echinococcus spp. prevalence. The potential contact between leashed dogs and wild small mammals is still a risk for re-infection of owned dogs with E. multilocularis. This study shows that the long-term application of regular dog treatment with praziquantel in the vast and remote echinococcosis endemic areas of the eastern Tibetan Plateau can reduce transmission in dogs but remains a challenging intervention.
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  • 文章类型: Evaluation Study
    Toxoplasma gondii infection is a worldwide parasitic zoonosis with a high-health risk for humans. The key epidemiological role played by felids is related to oocyst shedding. The present study compared two amplification protocols for the molecular diagnosis of Toxoplasma infection in owned cats. A total of 78 owned cats referred to an Italian university-teaching hospital and exposed to various T. gondii-associated risk factors were sampled for blood and faeces. Faecal specimens were processed by flotation and tested using 2 copro-PCRs targeting the widely used B1 gene and the 529-bp repetitive element (RE). The sera were tested by the indirect immunofluorescent antibody test (IFAT) for the detection of immunoglobulins against T. gondii. Sixteen faeces (20.52%) tested positive for T. gondii DNA; 12 samples were positive only at B1-PCR, two at 529-bp RE-PCR and two at both genetic targets (overall agreement = 82.11%). The amplicons obtained were sequenced, and the Basic Local Alignment Search Tool analysis showed a high homology with the T. gondii strains available in reference databases. Two stool samples were microscopically positive for T. gondii-like oocysts and also tested positive by both B1 and 529-bp RE-PCRs. Thirty-three (42.3%) sera tested positive for antibodies; of which, seven were found to have T. gondii DNA-positive results using the B1 genetic target (overall agreement = 57.77%). The amplification sets targeting B1 and 529-bp RE showed substantially different yields. Further research is needed to better understand the significance and the sensitivities of using these multi-copy-targeted molecular methods from cat faeces before being used for routine diagnosis.
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  • 文章类型: Journal Article
    Echinococcus granulosus (sensu lato) (s.l.) and Echinococcus vogeli are causative agents of chronic zoonotic diseases such as cystic and polycystic echinococcosis, respectively. In Brazil, polycystic echinococcosis has a restricted geographical distribution in the North Region, while cystic echinococcosis is observed in the South Region. Domestic dogs (Canis familiaris) fed with raw viscera represent a risk factor for E. granulosus (s.l.) infection in the South Region. Although this practice is frequent, it remains unclear whether domestic dogs are infected with E. vogeli in the state of Acre, located in the Amazon basin in the North Region of Brazil. The aim of this study was to investigate this gap in the polycystic echinococcosis epidemiology.
    Sixty-five fecal samples were collected from the ground in five municipalities (Sena Madureira, n = 14; Rio Branco, n = 06; Bujari, n = 06; Xapuri, n = 30; and Epitaciolândia, n = 09) located in the state of Acre, northern Brazil. The samples were screened for parasites by copro-PCR using the cox1 gene associated with automated sequencing.
    Echinococcus vogeli was molecularly confirmed in a sample from Sena Madureira and E. granulosus (sensu stricto) (s.s.) (G1) in a sample from Rio Branco.
    These findings indicate that molecular assays are useful in typing Echinococcus taxa from fecal samples of dogs in northern Brazil. The present study is the first molecular record of E. vogeli in domestic dogs found in the state of Acre, reinforcing their role as a source of infection for humans. Because E. granulosus (s.s.) (G1) was detected for the first time in the North Region, from the epidemiological standpoint this finding is highly relevant, because it expands the known geographical distribution, which was previously restricted to the South Region of Brazil.
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