Babesia DNA detection in canine blood and Dermacentor reticulatus ticks in southwestern Siberia, Russia

Babesia infection was studied in 21 blood samples of dogs with symptoms of babesiosis and among 72 Dermacentor reticulatus and 70 Ixodes persulcatus ticks from southwestern Siberia, Russia. Babesia DNA was detected by hemi-nested PCR based on the 18S rRNA gene with subsequent direct sequencing. All of the analyzed canine blood samples and three D. reticulatus, but none from I. persulcatus ticks studied were shown to contain Babesia DNA. Nucleotide sequences of the Babesia 18S rRNA gene fragment of 354 bp long for all 24 positive samples appeared to belong to the subspecies Babesia canis canis and differed only at three positions. The Babesia nucleotide sequences from 17 canine blood samples and from one D. reticulatus tick were identical to each other and to previously known B. canis canis from canine blood in Slovenia. Four canine blood samples and the second tick sample contained a mixture of two nucleotide sequences previously found in canine blood. B. canis canis nucleotide sequence from the third tick differed in the unique nucleotide transition and could correspond to a new genetic variant. Thus, the main etiological agent of canine babesiosis in Novosibirsk region is B. canis canis, and D. reticulatus, but not I. persulcatus, ticks could serve as a vector of this infectious agent. To our knowledge, this is the first report of the B. canis canis nucleotide sequences from ticks.     

Wild cervids are host for tick vectors of babesia species with zoonotic capability in Belgium

Babesiosis is a tick-borne disease caused by different species of intraerythrocytic protozoan parasites within the genus Babesia. Different species of Babesia are described as potentially zoonotic and cause a malaria-like disease mainly in immunocompromised humans. Interest in the zoonotic potential of Babesia is growing and babesiosis has been described by some authors as an emergent zoonotic disease. The role of cervids to maintain tick populations and act as a reservoir host for some Babesia spp. with zoonotic capability is suspected. To investigate the range and infection rate of Babesia species, ticks were collected from wild cervids in southern Belgium during 2008. DNA extraction was performed for individual ticks, and each sample was evaluated for the absence of PCR inhibition using a PCR test. A Babesia spp. genus-specific PCR based on the 18S rRNA gene was applied to validated tick DNA extracts. A total of 1044 Ixodes ricinus ticks were collected and 1023 validated samples were subsequently screened for the presence of Babesia spp. DNA. Twenty-eight tick samples were found to be positive and identified after sequencing as containing DNA representing: Babesia divergens (3), B. divergens-like (5), Babesia sp. EU1 (11), Babesia sp. EU1-like (3), B. capreoli (2), or unknown Babesia sp. (4). This study confirms the presence of potentially zoonotic species and Babesia capreoli in Belgium, with a tick infection rate of 2.7% (95% CI 1.8,3.9%). Knowledge of the most common reservoir source for transmission of zoonotic Babesia spp. will be useful for models assessing the risk potential of this infection to humans.     

First evidence of Babesia gibsoni (Asian genotype) in dogs in Western Europe

Canine babesiosis in Europe is generally caused by Babesia canis canis and Babesia canis rossi. Here we describe the first two autochthonous cases of Babesia gibsoni (Asian genotype) infection in Germany. Two American pit bull terriers showed clinical and hematologic signs consistent with babesiosis. Polymerase chain reaction (PCR) analysis of the 18S rDNA of blood samples revealed 486 bp fragments. The sequences were 100% identical to each other and 100% identical to Babesia gibsoni (Asian genotype). These results represent the first genetic evidence of Babesia gibsoni (Asian genotype) parasites in dogs in Western Europe.     

新疆克拉玛依地区蜱媒病原体的调查

目的 调查新疆克拉玛依地区蜱种类及其携带病原体。方法 于克拉玛依市不同区采集游离蜱和寄生蜱,鉴定蜱种类,利用荧光定量PCR或套氏PCR/半套氏PCR检测发热伴血小板减少综合征布尼亚病毒、斑点热群立克次体、克里米亚-刚果出血热病毒、蜱传脑炎病毒、巴贝斯虫/泰勒虫、贝氏立克次体、查菲埃立克次体、恙虫病东方体和伯氏疏螺旋体等9种病原体。结果 共采集蜱683只,分3属3种,其中亚东璃眼蜱数量(n=362)最多,其次为银盾革蜱(n=311)、图兰扇头蜱(n=10)。银盾革蜱中斑点热群立克次体的阳性率为3.2%(10/311),巴贝斯虫的阳性率为1.0%(3/311);亚东璃眼蜱中巴贝斯虫的阳性率为0.8%(3/362),泰勒虫阳性率为0.3%(1/362);其他病原体检测结果均呈阴性。PCR扩增阳性基因序列分析显示银盾革蜱携斑点热群立克次体阳性与劳氏立克次体最相关,银盾革蜱和亚东璃眼蜱携带驽巴贝斯虫。结论 新疆克拉玛依地区优势蜱为亚东璃眼蜱和银盾革蜱,银盾革蜱携带劳氏立克次体,银盾革蜱和亚东璃眼蜱携带驽巴贝斯虫,提示该地区存在斑点热、巴贝斯虫病的自然疫源地,因此有必要加强对这些蜱携疾病的防控。     

广西壮族自治区犬巴贝虫感染现状调查

目的 分析广西壮族自治区犬巴贝虫病的流行情况. 方法 在广西的玉林、北海、南宁、百色和河池等地区的17个采样点,用FTA试纸卡采集犬血.巢式PCR特异性扩增巴贝虫18s rDNA基因,纯化阳性样本的PCR产物、测序,与GenBank中的巴贝虫序列进行比对.应用Mega5.1软件构建系统发育树,分析系统发生关系. 结果 共采获87份犬血,其中阳性血样11份,感染率为12.64％ （11/87）.11份阳性血样的巴贝虫序列相同,与GenBank中的佛氏巴贝虫（Babesia canis vo-geli的同源性为98.1％.在邻接法构建的系统进化树上,犬血样中检测到的巴贝虫与佛氏巴贝虫同属一个分枝,系统发生关系近. 结论 广西犬血中检测到的巴贝虫与GenBank中的佛氏巴贝虫同源性高,为佛氏巴贝虫.     

人巴贝虫病的临床特征及诊疗研究进展

巴贝虫病是一种重要的蜱传人兽共患血液寄生虫病,在全球分布较广泛.对人致病的巴贝虫主要有田鼠巴贝虫、分歧巴贝虫、猎户巴贝虫和邓肯巴贝虫等12种.巴贝虫感染人体后主要临床症状为发热、头痛、寒战、肌痛和疲劳等,严重时可致死.巴贝虫病主要通过实验室检测结合临床特征、流行病学调查进行诊断,诊断技术主要包括血涂片镜检、血清学检测和...     

Babesia spp. infections transmitted through blood transfusion

Babesiosis in humans is caused by infection with various species of Babesia (Apicomplexa, Piroplasmida), mainly transmitted by an arthropod vector--Ixodes spp. ticks. This review will focus on blood transfusion as another mode of Babesia transmission, especially in endemic areas, as well as the impact of human babesiosis on transfusion medicine.     

Roe deer (Capreolus capreolus) and red deer (Cervus elaphus) as a reservoir of protozoans from Babesia and Theileria genus in north-western Poland

The species of genus Babesia and Thelieria are obligate intracellular pathogens that multiply in both vertebrate and invertebrate hosts. Some species of Babesia cause bovine babesiosis infecting erythrocytes of the cattle and wild ruminants, and undergo a complex developmental cycle in ticks which serve as biological vectors. Majority of Theileria spp. cause bovine theileriosis infecting lymphocytes as well as erythrocytes of the cattle and wild ruminants, and similar to Babesia undergo a complex developmental cycle in ticks. In this study, hunter killed roe deer (Capreolus capreolus) and red deer (Cervus elaphus) from north-western Poland were tested for Babesia and Theileria infection in two seasons (spring and autumn, 2004). Infection with babesias and theilerias was detected by PCR assay based on the fragment of nuclear small subunit rRNA gene (nss-ribosomal DNA). Four types of products different in size were obtained and then sequenced. Sequence analysis of nucleotides showed that two kinds of products (385 and 475 bp) were unspecific, the third was characteristic for Theileria sp. (430bp) and the last one for Babesia divergens (407bp). We found that 24.4% of the animals examined were infected with Babesia divergens and 11% with Theileria sp. Percentage of infected animals with B. divergens was almost equal in the spring and autumn (24.6 and 24% respectively). Infection with Theileria was lower in the spring than in the autumn (10.5 and 12% respectively).     

人巴贝虫病研究进展

巴贝虫病是巴贝虫在红细胞内寄生导致的一种人畜共患寄生虫病.人巴贝虫病主要由田鼠巴贝虫（Babesia microti）、B.duncani、分歧巴贝虫（B.divergens）、B.venatorum等引起,病例呈世界性分布.其传播方式有蜱虫叮咬、输血及母婴传播等.患病症状与疟疾相似,典型的临床症状表现为发热、肌痛、贫血、血红蛋白尿、黄疸等,其严重程度主要取决于自身免疫状况和感染巴贝虫的种类.巴贝虫病常见的诊断方法为涂片染色法、动物接种法、血清学检测法以及分子生物学检测法.     

Anaplasma phagocytophila and protozoans of Babesia genus in dogs from endemic areas of Lyme disease in north-western Poland

Infections caused by the spirochete Borrelia burgdorferi sensu lato may be accompanied by other microorganisms, such as Anaplasma, Ehrlichia and Babesia. These pathogens are transmitted by the ticks and are a risk to humans and animals. Ixodes ricinus ticks collected from recreational areas of Szczecin and northwestern Poland contained DNA of the pathogens mentioned above and cases of double and triple coinfection have been documented. The aim of this paper was to determine if dogs suspect to tick infestation in the area of study are a reservoir for these pathogens and to examine the possibility of coinfection. Canine blood was sampled, part of the material originated from dogs exhibiting symptoms of borreliosis. In an earlier study, the samples were screened for DNA from B. burgdorferi sensu lato. In order to screen for A. phagocytophila and Babesia sp. DNA, a PCR-based method was used with the following primers: EHR521/EHR747 for Anaplasma and FOR1/REV1 for Babesia. In 192 samples only two contained A. phagocytophila DNA. One of these samples originated from a healthy canine, the other from an individual with symptoms of borreliosis. The examined samples were not positive for Babesia sp. DNA. Coinfection was not discovered. The low level of A. phagocytophila infection may indicate that the domestic dog is not a reservoir for Anaplasma and Babesia in Szczecin and northwestern Poland. Moreover, this area does not have populations of the brown dog tick (Rhipicephalus sanguineus) or Dermacentor reticulates--both of which are vectors of E. canis and B. canis and commonly induce ehrlichiosis and babesiosis in canines.     

Survey for Tick-Borne Zoonoses in the State of Espirito Santo,Southeastern Brazil

Blood samples collected from 201 humans, 92 dogs, and 27 horses in the state of Espirito Santo, Brazil, were tested by polymerase chain reaction, indirect immunofluorescence assays, and indirect enzyme-linked immunosorbent assay for tick-borne diseases (rickettsiosis, ehrlichiosis, anaplasmosis, borreliosis, babesiosis). Our results indicated that the surveyed counties are endemic for spotted fever group rickettsiosis because sera from 70 (34.8%) humans, 7 (7.6%) dogs, and 7 (25.9%) horses were reactive to at least one of the six Rickettsia species tested. Although there was evidence of ehrlichiosis (Ehrlichia canis) and babesiosis (Babesia canis vogeli, Theileria equi) in domestic animals, no human was positive for babesiosis and only four individuals were serologically positive for E. canis. Borrelia burgdorferi-serologic reactive sera were rare among humans and horses, but encompassed 51% of the canine samples, suggesting that dogs and their ticks can be part of the epidemiological cycle of the causative agent of the Brazilian zoonosis, named Baggio-Yoshinari Syndrome.     

Wild ungulates as Babesia hosts in northern and central Italy

Babesia and Theileria species were investigated in wild ungulates of Northern and Central Italy. Of 355 blood samples examined, 108 (30.4%) were positive to molecular diagnostics (polymerase chain reaction [PCR] with specific primers and sequencing). The sequence analysis showed that the roe deer is a susceptible host for several piroplasms belonging both to Babesia (31%) and Theileria (14.2%) species, whereas fallow deer and wild boar harbor only Theileria species (49% and 2.6%, respectively). Strains related to B. divergens are highly present (28.3%) in the roe deer, which, however, also harbors Babesia MO1 type and Babesia microti-like organisms. Babesia EU1 type is described for the first time in a roe deer in Italy. The finding in roe deer of Babesia species involved in human babesiosis is of concern for public health, mainly because ecological changes in progress cause the increase of both the deer species and the vector tick populations.     

我国巴贝虫病流行现状与研究进展

巴贝虫病是由巴贝虫感染引起的一类人兽共患寄生虫病。巴贝虫经蜱叮咬、输血或器官移植等途径传播,主要寄生于人或其他脊椎动物红细胞内。感染人体的巴贝虫主要有田鼠巴贝虫、猎户巴贝虫、邓肯巴贝虫、分歧巴贝虫等。巴贝虫病呈世界性分布,多发于夏秋季节,在我国属于新发、罕见寄生虫病。过去临床上缺乏敏感有效的诊断方法,导致很多巴贝虫病患者未能得到及时治疗。随着高敏感性的分子生物学检测技术在临床上的应用,巴贝虫病病例报告数量逐渐增多。本文详细回顾了1943年至今我国人巴贝虫病病例报告情况,概述了巴贝虫病诊断、治疗和致病机制等方面的研究进展,分析了我国巴贝虫病防控的挑战和前景。     

Comparative infectivity of Babesia divergens and a zoonotic Babesia divergens-like parasite in cattle

Babesia divergens-like parasites identified in human babesiosis cases in Missouri and Kentucky and in eastern cottontail rabbits (Sylvilagus floridanus) on Nantucket Island, Massachusetts, share identical small subunit ribosomal RNA gene sequences. This sequence is 99.8% identical to that of Babesia divergens, suggesting that the U.S. parasite may be B. divergens, a causative agent of human and bovine babesiosis in Europe. Holstein-Friesian calves were inoculated with cultured Nantucket Island Babesia sp. (NR831) and B. divergens parasites and monitored by clinical signs, Giemsa-stained blood films, PCR, and culture. The NR831 recipients did not exhibit clinical signs of infection and remained negative for all assays. The B. divergens recipients developed clinical infections and became positive by all assays. NR831 recipients were fully susceptible upon challenge inoculation with B. divergens. This study confirms that the Nantucket Island Babesia sp. is not conspecific with B. divergens based on host specificity for cattle.     

Occurrence of Babesia microti in ticks Ixodes ricinus on selected areas of western Pomerania

The aim of present study was to evaluate acquisition risk of babesiosis in human population exposed to ticks Ixodes ricinus by examination of Babesia microti DNA occurrence in ticks of all development stages. The polymerase chain reaction (PCR) was applied to estimate the occurrence of DNA Babesia microti in Ixodes ricinus. The Bab1 and Bab4 primers were used to amplify fragment. 238bp in length, of 18S rRNA gene for small ribosomal subunit. Amplicons were electroforeticaly separated in agarose gels. Ticks were collected in year 1999 and 2000, twice in each year in spring-summer (May-July) and summer-autumn (August-October) seasons from Goleni6w Forest and Pobierowo. These places have been classified as people attendance and tourist areas. The 716 I. ricinus ticks were collected in 1999 with 61.3% of nymphs, 17.8% larvae, 10.9% females and 9.9% males. Highest range of infection was observed in females--28.8% studied, than males--18.3%, nymphs--7.7% and larvae--3.l1%. The total number of 416 I. ricinus was collected in year 2000 with 64% of nymphs. 13.4%/males. 11.9% females and 10.7% larvae. The infection with Babesia microtri occurred only in three nymphs, which was 0.7% of studied population.     

Detection of Borrelia duttonii,a tick-borne relapsing fever agent in central Tanzania,within ticks by flagellin gene-based nested polymerase chain reaction

Argasid ticks collected in the site near Mvumi Hospital, Dodoma, Tanzania, were subjected to flagellin gene-based nested polymerase chain reaction (PCR) amplification for examination of borrelial infections. Eight of 13 ticks gave a strong 350-bp signal; three had a weak signal at the same size, and the rest were negative. Sequence determination of eight of the positive samples resulted in three types of flagellin gene sequences. The first type of sequence (shown by three individuals) was identical to that of Borrelia duttonii strain Ly. The second type of sequence from PCR products from four individual ticks had only one base substitution without amino acid alteration in deduced protein sequence. The third type of sequence was different from that of any other Old World relapsing fever borreliae, and the tick was thought to be infected with an unknown Borrelia species. Ticks were also examined to determine the nucleotide sequence of the mitochondrial 16S rRNA gene. The partial sequence of approximately 470 bases was aligned for comparison with previously published sequences to identify the species. The sequences of 13 individual ticks were all identical, and the sequence similarity analysis revealed the ticks should be classified as members of the Ornithodoros porcinus species. The PCR method described in this report appears to be a reliable tool for the detection of borreliae and epidemiological study of tick-borne relapsing fever.     

福建省1例人巴贝虫病的诊断与鉴定

目的 分析1例人感染巴贝虫的实验室资料,提高对巴贝西虫病的诊断水平。方法 观察外周血中虫体的形态;从患者外周血中提取的DNA核酸,采用田鼠巴贝虫种特异性引物进行聚合酶链反应(PCR)。采用PCR扩增田鼠巴贝虫18S rRNA片段,取阳性PCR产物送测序并进行序列比对分析。结果 外周血中见大量疑似恶性疟原虫虫体;PCR产物测序结果经过BLAST比对,与田鼠巴贝虫18s核糖体DNA序列有99%的同源性。结论 感染的虫体为田鼠巴贝虫。福建省部分地区鼠类(尤其是野鼠)存在田鼠巴贝虫感染,是巴贝虫病的自然疫源地,必须引起足够的重视。     

黑河口岸地区蜱类调查及研究

目的 调查分析黑河口岸地区蜱种类多样性。方法 对2012年至2014年黑龙江省黑河区域内蜱种群密度、数量、分布以及季节变迁方面的数据进行整理及总结,通过形态学的观察及PCR检测明确黑河地区蜱的分类及携带病原体情况。结果 黑河口岸地区的蜱类采集活动中,共获得蜱类1 343只,经鉴定分属3属3种,其中森林革蜱1 054只,占总体构成78.48%,为优势蜱种;嗜群血蜱236只,占总体构成17.57%;全沟硬蜱53只,占总体构成3.94%。之后本试验对采集到的1 343只蜱针对10种病原体进行PCR检测。在黑河地区的蜱类中,森林革蜱的立克次体感染率为64.37%、巴尔通体感染率为3.35%、类似巴贝西虫病原体检出率为1.70%,其他相关蜱虫病原检测结果为阴性。结论 森林革蜱为试验采集蜱虫中的优势蜱种;进行病原体检查发现立克次体具有较高感染率,其中病原体主要在森林革蜱中发现,森林革蜱仍然是监测新发蜱媒传染病的前哨种类。携带的病原体的种类随着生态环境的变化在慢慢发生变化,边境的生态环境复杂多样,在蜱虫比较活跃繁多的地区和季节应该增大防范措施,以免人们在工作以及生活中被其感染,从而造成生命财产损失。     

Ticks and associated pathogens collected from domestic animals in the Netherlands

Following an outbreak of autochthonous canine babesiosis in the Netherlands, a request made to veterinarians and the public to collect ticks from companion animals resulted in 4298 ticks submitted between July 2005 and October 2006 to our center. Ticks were identified as Ixodes ricinus adults (2907/4298, 67.6%), Ixodes sp. nymphs (529/4298, 12.3%) and Ixodes sp. larvae (385/4298, 9.0%), I. hexagonus adults (328/4298, 7.6%), Dermacentor reticulatus (72/4298, 1.7%), and several other exotic tick species such as Amblyomma flavomaculatum (formerly Aponomma flavomaculatum), Hyalomma marginatum rufipes, Rhipicephalus sanguineus, and R. turanicus (55/4298, 1.3%). Eight localities were surveyed for the presence of local D. reticulatus, a tick not indigenous to the Netherlands, based on multiple submissions of D. reticulatus ticks from these areas. D. reticulatus was collected from the vegetation in six of these localities, confirming the presence of populations of this tick in the Netherlands. Adult I. ricinus (n=251), I. hexagonus (n=237), and D. reticulatus (n=344) ticks were selected at random and subsequently screened by polymerase chain reaction (PCR) and reverse line blot (RLB) hybridization for the presence of Borrelia, Babesia, Theileria, Anaplasma, Ehrlichia, and Rickettsia species. I. ricinus ticks were infected with Rickettsia helvetica (24.7%), spirochetes belonging to the Borrelia burgdorferi sensu lato group (7.2%), the Ehrlichia-like "Schotii" variant (2.4%), Anaplasma phagocytophilum (1.6%), Babesia sp. (EU1) (1.2%), Babesia divergens (0.4%), and Babesia microti (0.4%). A. phagocytophilum (5.9%) and R. helvetica (0.8%) were also detected in adult I. hexagonus ticks. Spotted fever group Rickettsiae, previously reported as Rickettsia sp. DnS14/RpA4 (14.0%), and Borrelia burgdorferi sensu lato (0.3%) were detected in the D. reticulatus ticks, which appeared to be free from B. canis infection. We concluded that a much broader spectrum of ticks and tick-borne pathogens is present in the Netherlands than previously thought, including several potential zoonotic pathogens.     

Circulation of Brucellaceae,Anaplasma and Ehrlichia spp. in borderline of Iran,Azerbaijan, and Armenia

Objective: To estimate the infection of ticks to Anaplasma, Ehrlichia, Babesia, Theileria, and Brucellaceae using molecular methods in borderline of Iran, Azerbaijan, and Armenia.Methods: Totally, 2 022 ticks were collected from different livestock. Then, species were diagnosed under stereomicroscope according to valid morphological keys. Tick DNA was extracted followed by PCR to detect Anaplasma, Ehrlichia, Theileria, Babesia and Brucellaceae infection in ticks. Results: A total of 498 males [24.62%(95% CI 22.76%-26.57%)], 741 females [36.64%(95% CI 34.54%-38.79%)], 782 nymphs [38.67%(95% CI 36.55%-40.84%)] and 1 larva [0.04%(95% CI 0.00%-0.28%)] were identified. Among identified samples, we found four genera including Hyalomma, Rhipicephalus, Haemaphysalis, and Dermacentor. Molecular assay revealed that the prevalence of ticks to Anaplasma or Ehrlichia, and Brucellaceae was 22.02%(95% CI 16.01%-29.06%) and 15.03%(95% CI 9.43%-22.26%), respectively. Phylogenetic analysis showed that the identified Anaplasma sp. had the most similarity with Anaplasma centrale, Anaplasma platys, Anaplasma camelii, and Anaplasma phagocytophilum, submitted in Gen Bank. Furthermore, the detected Ehrlichia sp. and Brucellaceae bacterium had the most similarity with Ehrlichia ruminantium and Mycoplana peli, respectively. However, no sign of the presence of Theileria and Babesia spp. was seen in the studied samples. Conclusions: Anaplasmosis, ehrlichiosis and brucellosis should be considered as important health threats in northwestern Iran and consistent monitoring on infection of ticks and livestock should be performed regularly.