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1.
At the abattoir on study in Miyazaki, Japan, 9537 imported cattle from Australia in average were slaughtered annually in the last 5 years (2006 to 2010) and hydatid cysts were constantly detected in about 1.8% of the cattle. In order to assess the risk of Echinococcus granulosus delivered to Japan by imported cattle, 250 cysts found in 103 cattle at the abattoir were examined for their biological characteristics and genotypes. The cattle slaughtered were imported from Australia at an age of 10-12 months old and fattened for 17-18 months in Japan. The cysts showed their size ranging from 4 to 108 mm and were mainly found in the lung. Mature protoscoleces were detected in the three largest cysts, all were of the G1 genotype. Most of the other cysts contained clear cyst fluid and had thin laminated layer with no protoscoleces. The finding implies a potential risk of E. granulosus being established in Japan, thus strict and proper meat inspection and consequent offal condemnation are requisite at abattoirs that deal with imported cattle. Genotyping based on partial fragments of mitochondrial cox1, rrnS and nad1 genes were performed on the 66 cysts, showing that most of the cysts were G1 genotype (common sheep strain). However, two and four cysts were considered as G2 (Tasmanian sheep strain) and G3 (buffalo strain) genotypes, respectively. Since it has been widely recognized that G1 is the only genotype distributing in mainland Australia and that G2 genotype has been eradicated from Tasmania, the finding of those genotypes from Australian cattle indicated that certain genotypes other than G1 genotype are distributing in mainland Australia.  相似文献   

2.
Garippa G 《Parassitologia》2006,48(1-2):57-59
An update on Cystic Echinococcosis (CE) diffusion in Italy during 2003-2005 is reported. CE seems to have a sporadic diffusion in the northern part of the country where this disease plays a minor role (prevalence < 1%). Recent investigations have shown the occurrence of CE cases in humans from the mountains between Reggio Emilia and Modena, with an average year incidence between 9.4 and 5.6/100,000. In Abruzzo prevalences in sheep and cattle are 20.2% and 15.3%, with a fertility of 4.6% and 1.3%, respectively. In the same region, G1 and G3 strains were identified and a prevalence of 31% in dogs was found with CaELISA. In Campania, CE prevalence was 14.8% in cattle, with no viable cysts recovered, and 10.5% in water buffaloes, with a fertility of 1.4%. Biotechnologies allowed to find G1 and G3 strains in water buffaloes. In Sicily, CE was found in 67.1% of cattle, with a fertility of 4%, and in 57.6% of sheep, with 9.2% of viable cysts. Biomolecular investigations have found G1 strain in sheep and cattle. In dogs, a prevalence of 5.6% for Echinococcus granulosus was reported. In Sardinia CE prevalence was 75.3% in sheep and 41.5% in cattle, with a fertility of 10.3% and 2.6%, respectively. CE was found also in 9.4% of pigs, with fertility of 6.5%. The G1 strain was recovered in sheep and cattle while the G7 in pigs.  相似文献   

3.
To facilitate ecotourism and research, free-ranging mountain gorillas of Uganda have been habituated to humans. Testing of fecal samples of gorillas (n = 100), people sharing gorilla habitats (n = 62). and local pre- and postweaned cattle (n = 50) having access to these habitats with fluorescein isothiocyanate-conjugated monoclonal antibodies revealed Giardia duodenalis cysts at prevalences of 2, 5, and 10%, respectively. The identification of G. duodenalis was confirmed by fluorescent in situ hybridization with 2 species-specific 18-bp oligonucleotide probes conjugated to hexachlorinated 6-carboxyfluorescein. The mean pathogen concentration was 2.5, 2.8, and 0.2 x 10(4) cysts/g of the gorilla, people, and cattle feces, respectively. All cyst isolates aligned with genotype (assemblage) A, as confirmed by polymerase chain reaction amplification and sequencing of a 130-bp region near the 5' end of the small subunit-ribosomal RNA gene. A single genotype (assemblage) A recovered from 3 genetically distant but geographically united host groups indicates anthropozoonotic transmission of G. duodenalis. A large percentage of the local community does not follow park regulations regarding the disposal of their fecal waste, as self-reported in a questionnaire. This genotype may have been introduced into gorilla populations through habituation activities and may have then been sustained in their habitats by anthropozoonotic transmission.  相似文献   

4.
A method for the isolation of Echinococcus granulosus DNA from germinal layers of hydatid cysts is described. The method includes a hexadecyltrimethylammonium bromide/chloroform extraction and an adsorption to diatomaceous earth suspension. DNA suitable for polymerase chain reaction was obtained and used for parasite strain determination by mitochondrial cytochrome c oxidase I gene sequencing. Fertile and nonfertile cyst isolates from sheep, cattle, pigs, and humans were characterized. Hitherto, no direct parasite strain characterization has been made on nonfertile hydatid cysts, whereas here we report that nonfertile hydatid cysts were produced by sheep strain (G1 genotype) in sheep, cattle, and humans and by pig strain (G7 genotype) in pigs.  相似文献   

5.
INTRODUCTION: Echinococcus granulosus species has a wide variety in both geography and hosts; indeed, 10 genotypes have been reported in studies on material of animal origin. The aim of this study was to genotype E. granulosus obtained from human hydatid cysts. MATERIALS AND METHODS: The hydatid fluid and sand was collected from patients who underwent surgery for hepatic and pulmonary hydatidosis at Hospital Regional in Temuco, Chile, between 2004 and 2005. Two PCR systems were used: PCR Eg 9 and PCR Eg 16. The RsaI enzyme was used for RFLP. The genotype was confirmed using the sequence of one fragment of 366 bp from a mitochondrial gene (cox1). RESULTS: The DNA of protoscolices from 24 samples was analyzed, 4 of them from pulmonary cysts and 20 from hepatic cysts. The 366 bp fragment was amplified in 20 out of 24 samples (83.3%). Enzymatic digestion revealed the presence of 3 possible genotypes: in 20 out of 21 samples (95,2%), a restriction was observed corresponding to the G1 or G7 genotypes; in the remaining sample genotype G4 or G7 was observed. Sequencing confirmed the presence of G1 genotype for 19 samples and G6 genotype for the remaining sample (G4 or G7 according to PCR-RFLP). CONCLUSION: The PCR-RFLP technique enabled three possible genotypes present (G1 or G7, G4 or G7) to be established. Sequencing allowed us to decisively identify the G1 and G6 genotypes in our study group. Previous studies agree with the identification of the G1 genotype in our country. We consider it significant that the G6 genotype is present in Chile for its epidemiological implications.  相似文献   

6.
运用同源序列克隆技术结合反转录PCR技术和3′,5′cDNA末端快速扩增技术得到了牛FABGL基因的完整CDS,3′非翻译区和部分5′非翻译区.序列分析和生物信息学研究表明,所获得的牛FABGL基因的cDNA包含994个核苷酸和780 bp的开放阅读框及198 bp的完整的3′非翻译区.该基因编码260个氨基酸残基蛋白,在氨基酸水平上与人的同源基因具有高度的相似性(88%).采用PCR-SSCP方法,在递交的包含完整CDS的长为1 925 bp的该基因的基因组DNA序列(GenBank接受号DQ409814)1 065 bp 和1 792 bp处,分别发现了两个单核苷酸碱基突变Y=C/T,R=A/G;它们分别位于该基因的第五和第八内含子.对包含这两个多态位点的3个品种(安格斯、海福特和西门塔尔)牛的共179个个体等位基因频率与部分肉质及生长性状进行了关联分析,结果发现,在第八内含子内具有GG基因型的个体的肉用性能指数(4.283±.0.475kg/cm)较具有AA基因型个体的(4.008±0.465kg/cm)高(P≤0.01);而且同一位点具有GG基因型的个体的眼肌面积(73.380±13.005 cm2)显著高于具有AA基因型的个体(67.744±12.777 cm2)(P≤0.05).在第五内含子内,具有CC、CT、TT3种不同基因型的个体之间,平均日增重差异均达到极显著水平(P≤0.01),以具有TT基因型的个体平均日增重最高(0.652±0.330kg/d),CC基因型的最低(0.421±0.178kg/d).  相似文献   

7.
Hydatid cyst caused by Echinococcus granulosus is one of the most important parasitic diseases around the world and many countries in Asia, including Iran, are involved with this infection. This disease can cause high mortality in humans as well as economic losses in livestock. To date, several molecular methods have been used to determine the genetic diversity of E. granulosus. So far, identification of E. granulosus using real-time PCR fluorescence-based quantitative assays has not been studied worldwide, also in Iran. Therefore, the aim of this study was to investigate the genetic diversity of E. granulosus from center of Iran using real-time PCR method. A total of 71 hydatid cysts were collected from infected sheep, goat, and cattle slaughtered in Isfahan, Iran during 2013. DNA was extracted from protoscolices and/or germinal layers from each individual cyst and used as template to amplify the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (420 bp). Five cattle isolates out of 71 isolates were sterile and excluded from further investigation. Overall, of 66 isolates, partial sequences of the cox1 gene of E. granulosus indicated the presence of genotypes G1 in 49 isolates (74.2%), G3 in 15 isolates (22.7%), and G6 in 2 isolates (3.0%) in infected intermediate hosts. Sixteen sequences of G1 genotype had microgenetic variants, and they were compared to the original sequence of cox1. However, isolates identified as G3 and G6 genotypes were completely consistent with original sequences. G1 genotype in livestock was the dominant genotype in Isfahan region, Iran.  相似文献   

8.
Echinococcus granulosus, the aetiologic agent of cystic echinococcosis (CE), is one of the most important zoonotic helminthes worldwide. Isolates of the parasite show considerable genetic variation in different intermediate hosts. Several genotypes and species are described in different eco-epidemiological settings. This study investigated E. granulosus genotypes existing in livestock and humans from the province of Kerman, located in south-eastern Iran, using sequencing data of cox1 and nad1 mitochondrial genes. Fifty-eight E. granulosus isolates, including 35 from sheep, 11 from cattle, 9 from camels and 3 from goats, were collected from slaughterhouses throughout Kerman. One human isolate was obtained from a surgical case of CE. Mitochondrial cox1 and nad1 regions were amplified using polymerase chain reaction (PCR) and 38 isolates were sequenced. Genotypes G1 (73.7%), G3 (13.2%) and G6 (13.1%) were identified from the isolates. G1 was the most common genotype from sheep (86.7%), cattle (80%), camels (44.4%) and goats (100%). Sheep, cattle and camels were also found to be infected with the G3 genotype (buffalo strain). The human isolate was identified as the G6 genotype. Results showed that the G3 genotype occurred in different animal hosts in addition to G1 and G6 genotypes.  相似文献   

9.
Climatic factors and on-farm management practices were evaluated for their association with the concentrations (cyst/liter) and instantaneous loads (cysts/second) of Giardia duodenalis in storm-based runoff from dairy lots and other high-cattle-use areas on five coastal California farms over two storm seasons. Direct fluorescent antibody analysis was used to quantitate cysts in 350 storm runoff samples. G. duodenalis was detected on all five dairy farms, with fluxes of 1 to 14,000 cysts/liter observed in 16% of samples. Cysts were detected in 41% of runoff samples collected near cattle less than 2 months old, compared to 10% of runoff samples collected near cattle over 6 months old. Furthermore, the concentrations and instantaneous loads of cysts were > or =65 and > or =79 times greater, respectively, in runoff from sites housing young calves than in sites housing other age classes of animals. Factors associated with environmental loading of G. duodenalis included cattle age, cattle stocking number, and precipitation but not lot area, land slope, or cattle density. Vegetated buffer strips were found to significantly reduce waterborne cysts in storm runoff: each additional meter of vegetated buffer placed below high-cattle-use areas was associated with reductions in the concentration and instantaneous load of cysts by factors of 0.86 and 0.79 (-0.07 and -0.10 log(10)/m), respectively. Straw mulch, seed application, scraping of manure, and cattle exclusion did not significantly affect the concentration or load of G. duodenalis cysts. The study findings suggest that vegetated buffer strips, especially when placed near dairy calf areas, should help reduce the environmental loading of these fecal protozoa discharging from dairy farms.  相似文献   

10.
In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17-22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin.  相似文献   

11.

Background

Cystic echinococcosis (CE) caused by the Echinococcus granulosus, is a major public health problem worldwide, including India. The different genotypes of E. granulosus responsible for human hydatidosis have been reported from endemic areas throughout the world. However, the genetic characterization of E. granulosus infecting the human population in India is lacking. The aim of study was to ascertain the genotype(s) of the parasite responsible for human hydatidosis in North India.

Methodology/Principal Findings

To study the transmission patterns of E. granulosus, genotypic analysis was performed on hydatid cysts obtained from 32 cystic echinococcosis (CE) patients residing in 7 different states of North India. Mitochondrial cytochrome c oxidase subunit1 (cox1) sequencing was done for molecular identification of the isolates. Most of the CE patients (30/32) were found to be infected with hydatid cyst of either G3 (53.1%) or G1 (40.62%) genotype and one each of G5 (cattle strain) and G6 (camel strain) genotype.

Conclusions/Significance

These findings demonstrate the zoonotic potential of G1 (sheep strain) and G3 (buffalo strain) genotypes of E. granulosus as these emerged as predominant genotypes infecting the humans in India. In addition to this, the present study reports the first human CE case infected with G5 genotype (cattle strain) in an Asian country and presence of G6 genotype (camel strain) in India. The results may have important implications in the planning of control strategies for human hydatidosis.  相似文献   

12.
Feces of harbor seals (Phoca vitulina richardsi) and hybrid glaucous-winged/western gulls (Larus glaucescens / occidentalis) from Washington State's inland marine waters were examined for Giardia and Cryptosporidium spp. to determine if genotypes carried by these wildlife species were the same genotypes that commonly infect humans and domestic animals. Using immunomagnetic separation followed by direct fluorescent antibody detection, Giardia spp. cysts were detected in 42% of seal fecal samples (41/97). Giardia-positive samples came from 90% of the sites (9/10) and the prevalence of positive seal fecal samples differed significantly among study sites. Fecal samples collected from seal haulout sites with over 400 animals were 4.7 times more likely to have Giardia spp. cysts than samples collected at smaller haulout sites. In gulls, a single Giardia sp. cyst was detected in 4% of fecal samples (3/78). Cryptosporidium spp. oocysts were not detected in any of the seals or gulls tested. Sequence analysis of a 398 bp segment of G. duodenalis DNA at the glutamate dehydrogenase locus suggested that 11 isolates originating from seals throughout the region were a novel genotype and 3 isolates obtained from a single site in south Puget Sound were the G. duodenalis canine genotype D. Real-time TaqMan PCR amplification and subsequent sequencing of a 52 bp small subunit ribosomal DNA region from novel harbor seal genotype isolates showed sequence homology to canine genotypes C and D. Sequence analysis of the 52 bp small subunit ribosomal DNA products from the 3 canine genotype isolates from seals produced mixed sequences at could not be evaluated.  相似文献   

13.
The occurrence of Thelazia gulosa and T. skrjabini in cattle in Italy, together with an update of the prevalence of Thelazia spp. in the southern Italy, is reported. On 764 slaughtered native cattle, fifty-five (7.2%) bovines were infected by Thelazia spp: T. rhodesi was found in 44 (80%) animals, T. gulosa was found in 19 (34.5%) cattle and only one (1.8%) cattle harboured T. skrjabini. This is the first report of T. gulosa and T. skrjabini in Southern Europe.  相似文献   

14.
Toll-like receptor 2 (TLR2) plays an important role in the innate immune response to a variety of pathogens. In this study, bovine TLR2 gene was taken as a candidate gene for mastitis resistance. Through PCR-SSCP analysis and sequencing, three missense mutations at T385 G, G398A, and G1884A were detected in the coding region that encoded extracellular domain. Altogether 240 dairy cattle of three breeds (Holstein, Simmental, and Sanhe cattle) were genotyped and allele frequencies were determined. The effects of TLR2 polymorphisms on somatic cell score (SCS) were analyzed and significant association was found between T385 G and SCS. The mean of genotype GG was significantly lower than those of genotype TT and TG. No significant associations were found with SCS for G398A and G1884A. Information provided in this research will be useful in further studies to determine the role of TLR2 gene in the mastitis resistance.  相似文献   

15.
BACKGROUND: Although the infection of Theileria sergenti is demonstrated by intraerythrocytic localization of this parasite, much time and labor are necessary in order to determine this. We applied flow cytometry to evaluate T. sergenti parasitemia using the fluorescent nucleic acid stain method. METHODS: Peripheral blood samples from cattle infected with T. sergenti were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16, and hydroethidine (HE). Stained parasitized erythrocytes were measured by a flow cytometer equipped with a single argon laser operating at 488 nm. RESULTS: SYTO16-stained intraerythrocytic parasites were detected on the FL1 (525 nm) and parasitized cells were separated completely from unparasitized cells. However, HE-stained erythrocytes could not be divided clearly into parasitized and unparasitized cells. SYTO16-stained parasites were reproducibly detected at a percentage above 0.1%. Contaminating leukocytes, which were indicated by CD18-positive cells, were eliminated from the analysis by narrowing the light scatter gate of the erythrocyte fraction. A correlation (r = 0.983) between the percentage of SYTO16-positive cells and parasitemia in grazing cattle was observed. CONCLUSIONS: Flow cytometric detection using SYTO16 is a rapid and reliable method of monitoring parasitemia in T. sergenti-infected cattle.  相似文献   

16.
He H  Liu X  Gu Y  Liu Y  Yang J 《Molecular biology reports》2011,38(8):4965-4969
In our study, genetic variation in coding region of cattle CCAAT enhancer binding protein alpha(namely CEBPA)gene was detected by PCR-RFLP and DNA sequencing methods in 215 individuals from Qinchuan cattle breed. Two haplotypes (A and B) and three observed genotypes (AA, AB, and BB) were detected. The result of DNA sequence showed one mutation by comparisons with NC_007316. The mutation at nt963 (T>G) were located in coding region of the CEBPA gene. Associations between the CEBPA gene genetic variation and the carcass traits were revealed in Qinchuan cattle. Least squares analysis revealed a significant statistical effect of the CEBPA gene different genotypes on slaughter weight and carcass weight in Qinchuan cattle. Individuals with BB genotype showed higher slaughter weight and carcass weight than individuals with AA and AB genotypes. Therefore, these results suggest that the CEBPA gene is a strong candidate gene that affects carcass traits in Qinchuan cattle.  相似文献   

17.
通过室内试验测定长枝木霉分生孢子悬浮液对小麦禾谷胞囊线虫胞囊的寄生和致死作用.结果表明: 不同浓度(1.5×105~1.5×107 cfu·mL-1)长枝木霉分生孢子悬浮液对小麦禾谷胞囊线虫胞囊具有明显的寄生和致死作用,并且不同浓度的长枝木霉分生孢子悬浮液之间存在显著差异.第18天浓度为1.5×107 cfu·mL-1的长枝木霉分生孢子悬浮液对胞囊的寄生率为96.7%,第22天对胞囊孵化的相对抑制率为91.2%.显微观察表明,侵染初期长枝木霉分生孢子附着在胞囊体表,并且萌发产生大量的菌丝寄生于胞囊体表,使〖JP2〗胞囊胚胎发育停止和内容物凝集,甚至有的胞囊出现畸形和表面形成深褐色的小液泡.侵染后期大量菌丝穿透胞囊体表,胞囊破裂,内容物外渗,有的胞囊体表菌丝形成分生孢子梗,其上着生卵圆形的分生孢子.表明长枝木霉可作为一种高效的生防制剂防治小麦禾谷胞囊线虫的发生与危害.  相似文献   

18.
The complete CDS sequence of the bovine FABGL gene was determined by homology cloning approach combined with RT-PCR and 3′- and 5′-RACE. The results of sequence analysis and bioinformatics study showed that this cDNA contained 994 nucleotides, with a 780 bp open reading frame (ORF) flanked by a 16 bp 5′-UTR (incompletely) and a 198 bp 3′-UTR. The deduced amino acid sequence (260 AA) shows 88% identity with the corresponding sequence in humans. Two single nucleotide substitutions, one located in intron 5 (I5) at position 1 065 bp (Y = C/T) (GenBank: DQ409814) and the other in intron 8 (I8) at position 1 792 bp (R = A/G), were detected using the PCR-SSCP method. Analysis of the allele frequencies of the two polymorphic sites in three different cattle breeds (Angus, Hereford, and Simmental) with different genotypes showed large differences: in locus I8, cattle with the GG genotype showed higher beef performance index (BPI) (4.283 ± 0.475 kg/cm) in comparison with cattle with the AA genotype (4.008 ± 0.465 kg/cm) (P = 0.01). Regarding the ribeye area, cattle with the GG genotype showed significantly higher ribeye area (73.380 ± 13.005 cm2) compared with cattle with the AA genotype (67.744 ± 12.777 cm2) (p = 0.05). In locus I5, some associations for the average daily gain (ADG) were found at the significance level of 0.01 between three different genotypes (CC, CT, TT): cattle with the TT genotype showed the highest ADG (0.652 ± 0.330 kg/d), whereas cattle with the CC genotype showed the lowest ADG value (0.421 ± 0.178 kg/d).  相似文献   

19.
Soba B  Logar J 《Parasitology》2008,135(11):1263-1270
To assess the importance of cattle as a source of human cryptosporidial infections in Slovenia, Cryptosporidium isolates from calves and humans with cryptosporidiosis were characterized genetically by direct DNA sequencing, targeting a variable region of the 60 subtypes', were identified, of which 7 were novel. In humans, C. hominis Ia (subtype IaA17R3) and Ib (IbA10G2) and Cryptosporidium parvum IIa (IIaA9G1R1, IIaA11G2R1, IIaA13R1, IIaA14G1R1, IIaA15G1R1, IIaA15G2R1, IIaA16G1R1, IIaA17G1R1 and IIaA19G1R1), IIc (IIcA5G3), and IIl (IIlA16R2) were recorded; this is the first record of the latter subtype in humans. In cattle, C. parvum IIa (IIaA13R1, IIaA15G2R1, IIaA16R1 and IIaA16G1R1) and IIl (IIlA16R2 and IIlA18R2) were recorded. Of the 15 subtypes identified, subtypes of C. parvum IIa were the most frequently encountered (>90%) in both humans and calves. The present findings suggest that zoonotic transmission plays an important role in sporadic human cryptosporidiosis in Slovenia.  相似文献   

20.
Cystic echinococcosis is a zoonotic parasitic disease caused by Echinococcus species. Tanzania is one of the endemic countries with cystic echinococcosis. This study focussed on identifying genotypes of Echinococcus spp. in Tanzania. We collected 7 cysts from cattle in Mwanza municipal (n=4) and Loliondo district (n=3). The cysts from Mwanza were all E. ortleppi and fertile. In contrast, the cysts from Loliondo were all E. granulosus sensu stricto and sterile. Two from the 4 cysts were a new haplotype of E. ortleppi (G5). These results can improve the preventive and control programs for humans and livestock in Tanzania. To our knowledge, this study is considered the first to identify the genotype and haplotype of Echinococcus spp. in Tanzania.  相似文献   

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