Agents In Ticks Research Articles

Identificación de Rickettsia andeanae en dos regiones de Chile

Candidatus 'Rickettsia andeanae' is recently recognized specie, with no attributed pathogenic role so far. It has been reported in Amblyomma ticks from Peru, USA and Argentina, and recently in Chile, in one specimen of Amblyomma triste from a rural locality in Arica. To determine the presence of rickettsial agents in ticks from domestic dogs from two Chilean regions. A transversal descriptive study in Coquimbo and La Araucanía regions, consisted in households sampling in the cities of Coquimbo and Aingol and nearby rural localities, in spring-summer 2011-2012. One dog per house was examined and ticks were collected when present; after taxonomic identification, gltA and ompA genes were amplified and sequenced. 462 dogs were examined, 255 had ticks (55%). In the cities just Rhipicephalus sanguineus was found; in rural localities also Amblyomma tigrinum. Obtained sequences in 12 specimens from A. tigrinum corresponded to Candidatus 'Rickettsia andeanae'. We documented the presence of R. andeanae in two distant Chilean regions, which is added to its recently finding in the Northern extreme of the country. This new rickettsial agent limits to Amblyomma ticks and rural areas, as it was reported in other American countries and corresponds with this tick habitat.

Do pathogens contribute to multiple sclerosis aetiology?

performance and interpretation from the Second National Conference on Serologic Diagnosis of Lyme Disease. MMWR 44, 590–591. 20. Russell, R.C. et al. (1994) Lyme disease: a search for a causative agent in ticks in south-eastern Australia. Epidemiol. Infect. 112, 375–384. doi:10.1017/ S0950268800057782 21. Marques, A. (2008)Chronic Lymedisease: a review. Infect.Dis. Clin.NorthAm.22, 341–360. [vii–viii] doi:10.1016/j.idc.2007.12.011 22. Krupp, L.B. et al. (2003) Study and treatment of post Lyme disease (STOP-LD): a randomized double masked clinical trial. Neurology 60, 1923–1930. doi:10.1212/ 01.WNL.0000071227.23769.9E 23. Fallon, B.A. et al. (2008) A randomized, placebo-controlled trial of repeated IV antibiotic therapy for Lyme encephalopathy. Neurology 70, 992–1003. doi:10.1212/01.WNL.0000284604.61160.2d 24. Vollmer-Conna, U. et al. (2007) Post-infective fatigue syndrome is not associated with altered cytokine production. Clin. Infect. Dis. 45, 732–735. doi:10.1086/ 520990 25. Cameron,B. etal. (2010) Serumcytokine levels inpost-infective fatigue syndrome. Clin. Infect. Dis. 50, 278–279. doi:10.1086/649546 Biography Professor Andrew Lloyd AM, BBS, MD, FRACP is an infectious diseases physician and immunology researcher. He leads a multidisciplinary teamof clinicians and scientists in research studying the biological basis of inflammation in human infectious diseases, including studies of hepatitis C infection and post-infective fatigue states. He is a NHMRC Practitioner Fellow and his research is supported by a NHMRC Program, Partnership and Project Grants. He is the Director of Infection and Inflammation Research Centre (IIRC) in the School of Medical Sciences at the University of New South Wales. Professor Lloyd was awarded an Australia Medal (AM) for his research achievements in infectious diseases and for hiswork in establishing the hepatitis service in the NSW prisons.

Ixodid fauna and zoonotic agents in ticks from dogs: first report of Rickettsia rickettsii in Rhipicephalus sanguineus in the state of Mato Grosso do Sul, mid-western Brazil

Ticks from 148 dogs from the urban area of the municipality of Campo Grande, state of Mato Grosso do Sul, Brazil, were collected, classified and analyzed using polymerase chain reaction (PCR) for the identification of Rickettsia spp., Trypanosoma cruzi and Leishmania spp. A total of 2015 ticks were collected. The species Rhipicephalus sanguineus (98.9%) and Amblyomma cajennense (1.1%) were identified. Molecular analysis revealed that no tick samples were infected by T. cruzi. Regarding Leishmania spp., tick samples from 36 dogs spread across all regions of the municipality were positive for L. chagasi. One tick sample was positive for Rickettsia spp. (gltA gene) in the PCR reaction. This sample was submitted to further PCR based on the ompA gene and the amplicon was sequenced. Identity of 100% was found with homologous sequences of R. rickettsii available in GenBank. This paper is the first to report the natural infection of R. sanguineus by R. rickettsii in the municipality of Campo Grande, state of Mato Grosso do Sul, mid-western Brazil.

Molecular detection of rickettsial agents in ticks and fleas collected from a European hedgehog (Erinaceus europaeus) in Marseilles, France

The European hedgehog (Erinaceus europaeus) is a synanthropic nocturnal insectivore commonly found in the countryside and in the parks and gardens. Because hedgehogs are already involved in the transmission of a number of zoonoses, including salmonellosis and ringworm, we decided to study their possible role in the epidemiology of the spotted fever group of Rickettsia. We collected ticks and fleas from a hedgehog that was captured in the city of Marseilles in France. Using a genus-specific quantitative PCR (qPCR) analysis followed by a species-specific qPCR analysis for positive samples, we observed that 91.7% (11/12) of the Rhipicephalus sanguineus ticks were positive for Rickettsia massiliae and 99.2% (128/129) of the Archaeopsylla erinacei fleas were positive for Rickettsia felis. Hedgehogs carry infected ectoparasites and then likely ensure the dissemination of spotted fever group Rickettsiae, and their epidemiological role requires further investigation.

Absence of Rickettsia rickettsii and Occurrence of Other Spotted Fever Group Rickettsiae in Ticks from Tennessee

Rocky Mountain spotted fever (RMSF) is the most common tick-borne illness in Tennessee. Little is known about the occurrence of R. rickettsii, the causative agent, in ticks in Tennessee. To better understand the prevalence and distribution of rickettsial agents in ticks, we tested 1,265 Amblyomma, Dermacentor, and Ixodes adult and nymphal ticks. Additionally, we tested 231 Amblyomma americanum larvae. Ticks were collected from 49 counties from humans, wild animals, domestic canines, and flannel drags. Spotted fever group rickettsiae (SFGR) DNA was detected by polymerase chain reaction (PCR) in 32% of adult and nymphal ticks. A total minimum infection rate of 85.63 was found in larval pools tested. Three rickettsial species, Rickettsia montana, Rickettsia amblyommii, and Rickettsia cooleyi were identified by molecular analysis. Rickettsia rickettsii was not detected. This study suggests that some RMSF cases reported in Tennessee may be caused by cross-reactivity with other SFGR antigenically related to R. rickettsii.

Borrelia,Ehrlichia, andRickettsiaspp. in Ticks Removed from Persons, Texas, USA

Data regarding the type, frequency, and distribution of tick-borne pathogens and bacterial agents are not widely available for many tick species that parasitize persons in the southern United States. We therefore analyzed the frequency and identity of pathogens and bacterial agents in ticks removed from humans and subsequently submitted to the Texas Department of State Health Services, Zoonosis Control Program, from October 1, 2004, through September 30, 2008. The data showed associations of bacterial agents and potential vectors. Tick-related illnesses may pose unidentified health risks in areas such as Texas, where incidence of human disease related to tick bites is low but well above zero and where ticks are not routinely suspected as the cause of disease. Cause, treatment, and prevention strategies can be better addressed through collecting sufficient data to establish baseline assessments of risk.

Detection and Identification of Rickettsial Agents in Ticks From Domestic Mammals in Eastern Panama

Several outbreaks of Rocky Mountain spotted fever have occurred in recent years in Colombian communities close to the border with Panama. However, little is known about rickettsiae and rickettsial diseases in eastern Panamanian provinces, the Darien Province and the Kuna Yala, located north of the endemic area in Colombia. In 2007, 289 ticks were collected in several towns from dogs, horses, mules, cows, and pigs. DNA was extracted from 124 Dermacentor nitens, 64 Rhipicephalus sanguineus, 43 Amblyomma ovale, 35 A. cajennense, 10 Boophilus microplus, 4 A. oblongoguttatum, and 9 A. cajennense nymphs. SYBR-Green polymerase chain reaction assays targeting a fragment of the OmpA and 16S rRNA genes were used for detection of DNA of the spotted fever group rickettsiae (SFGR) and Anaplasmataceae (Anaplasma and Ehrlichia), respectively. In total, 37.4% ticks were positive for SFGR, including 20.3% R. sanguineus, 27.9% A. ovale, 25.8% D. nitens, 50% B. microplus, 50% A. oblongoguttatum, and 100% A. cajennense. The presence of Rickettsia amblyommii DNA was confirmed by sequencing in A. cajennense, A. oblongoguttatum, A. ovale, B. microplus, and R. sanguineus. DNA of R. rickettsii was only detected in one D. nitens collected from a horse in Santa Fe, Darien Province. Prevalence of Anaplasmataceae varied from 6.3% in R. sanguineus to 26.5% in A. cajennense. DNA of Ehrlichia chaffensis was found in three D. nitens and three A. cajennense from horses. This is the first study providing molecular characterization and prevalence information on SFGR in ticks from these areas and thus will be helpful for future evaluations of the risk of rickettsial diseases for individuals living in this region.

Detection and Identification of the Spotted Fever Group Rickettsial Agents from Haemaphysalis Ticks in Jeju Island, Korea

This study investigated the presence of nucleic acids of various Rickettsial agents in ticks collected in Jeju Island, Korea from June 2007 to August 2008, through the nested polymerase chain reaction (PCR) and sequencing analysis of partial citrate synthase (gltA), Rickettsial outer membrane protein B (ompB), and 17-kDa genes. Examination of the 1,584 ticks showed that the subspecies distribution of Haemaphysalis longicornis was 99.81% (n=1,581) and H. flava was 0.19% (n=3). A total 224 out of 250 pools from one to 15 ticks were found to be positive in ompB-PCR assay (minimal infection rate 141 ticks/1,000 tested). From the positive samples, 26 were analyzed by gltA- and 17-kDa-PCR assays. The nucleotide sequences of the ompB- and gltA-PCR products showed a high degree of similarity with those of the Rickettsia japonica (98.7~99.2% and 98.7~99.3%, n=25) and R. monacensis (99% and 99.7%, n=1). However, analysis of the nucleotide sequences of the 17-kDa-PCR amplicons showed that the sequences of the 25 PCR amplicons were more close to R. marmionii (99.4~100%) than R. japonica (98.6~99.1%). These findings suggest that various rickettsial diseases could be transmitted via the bite of tick vectors in Jeju Island, Korea.

Differential diagnostics and special specific prophylaxis of tick-borne infections in Irkutsk city

The results of differential express diagnostic of tick-borne encephalitis and Lyme borreliosis agents in ticks, removed from pa- tients are present in this article (1995—2004). The entire complex of serolological and genetical methods were used. The specific prophylaxis based on the results of diagnostic had shown to be very effective for both infections. The first data about human ehrli- chiosis and anaplasmosis in Irkutsk region had been obtained. The improvement of diagnostic and prophylactic procedures are pro- posed.

Detection of <I>Anaplasma phagocytophilum</I> and <I>Babesia odocoilei</I> DNA in <I>Ixodes scapularis</I> (Acari: Ixodidae) Collected in Indiana

The blacklegged tick, Ixodes scapularis Say, first reported in Indiana in 1987, has now been detected in more than half of Indiana's counties. The first case of human granulocytic ehrlichiosis (human anaplasmosis) in Indiana was reported in 2002. We now report the detection of Anaplasma phagocytophilum and Babesia odocoilei (Emerson and Wright 1968) in I. scapularis ticks collected in northern Indiana. Using polymerase chain reaction analysis, 41 of 193 adult ticks (21.2%) collected from deer were positive for A. phagocytophylum, and 22 (11.4%) were positive for Babesia sp. Restriction fragment analysis of 12, and sequencing of another five of the amplified products identified these parasites as B. odocoilei. Five ticks (2.6%) were coinfected. Eight of 68 questing adult ticks (11.8%) were positive for A. phagocytophilum; seven (10.3%) were positive for Babesia sp. Six of the latter seven positive samples were determined to be B. odocoilei by restriction fragment analysis and sequencing of two samples. None of 39 pools of nymphs was positive for Babesia sp. Three of 15 ticks (20%) collected from a dog were positive for A. phagocytophilum and three ticks (20%) were positive for Babesia sp. One was confirmed as B. odocoilei. One tick was coinfected. This is the first report of the presence of these two agents in ticks in Indiana.

Detection of Anaplasma phagocytophilum and Babesia odocoilei DNA in Ixodes scapularis (Acari: Ixodidae) Collected in Indiana

Abstract The blacklegged tick, Ixodes scapularis Say, first reported in Indiana in 1987, has now been detected in more than half of Indiana’s counties. The first case of human granulocytic ehrlichiosis (human anaplasmosis) in Indiana was reported in 2002. We now report the detection of Anaplasma phagocytophilum and Babesia odocoilei in I. scapularis ticks collected in northern Indiana. Using polymerase chain reaction analysis, 41 of 193 adult ticks (21.2%) collected from deer were positive for A. phagocytophylum, and 22 (11.4%) were positive for Babesia sp. Restriction fragment analysis of 12, and sequencing of another five of the amplified products identified these parasites as B. odocoilei. Five ticks (2.6%) were coinfected. Eight of 68 questing adult ticks (11.8%) were positive for A. phagocytophilum; seven (10.3%) were positive for Babesia sp. Six of the latter seven positive samples were determined to be B. odocoilei by restriction fragment analysis and sequencing of two samples. None of 39 pools of nymphs was positive for Babesia sp. Three of 15 ticks (20%) collected from a dog were positive for A. phagocytophilum and three ticks (20%) were positive for Babesia sp. One was confirmed as B. odocoilei. One tick was coinfected. This is the first report of the presence of these two agents in ticks in Indiana.

Pathogens and symbionts in ticks: prevalence of Anaplasma phagocytophilum (Ehrlichia sp.), Wolbachia sp., Rickettsia sp., and Babesia sp. in Southern Germany.

Tick-transmitted diseases like tick-borne encephalitis and Lyme borreliosis have been well known in Germany for decades. Ongoing research now gives an additional focus to a broad range of other bacteria and parasites in ticks like Anaplasma phagocytophilum, former Ehrlichia sp., Rickettsia sp. and Babesia sp. Knowledge about the prevalence of these infectious agents in ticks is an important prerequisite for risk assessment of human diseases. Therefore nymphs and adult Ixodes ricinus ticks were collected and examined for Anaplasma phagocytophilum (n = 5424 ticks), Rickettsia sp. (n = 1187), and Babesia sp. (n = 3113). For the detection of Anaplasma phagocytophilum, DNA from the 16S rDNA gene was amplified by nested PCR and hybridized with a DIG-labeled oligonucleotide probe. The examination of Rickettsia sp. was performed by single PCR. A partial sequence of the citrate synthase gene was amplified. As a target for the detection of Babesia sp., DNA from the 18S rDNA gene was amplified, also by single PCR. All positive PCR products were sequenced to control specificity. Anaplasma phagocytophilum was detected by PCR in n = 103 (1.9%) out of 5,424 examined ticks from 11 investigation areas. However, not all positive PCR products hybridized using DIG-labeled oligonucleotide probe. Thus, the result of sequencing indicated that only 1.0% (n = 54) belonged to Anaplasma phagocytophilum and nearly half of these PCR products (0.9%) were identified as Wolbachia sp. Rickettsia sp. in Ixodes ricinus ticks from 3 areas were found in n = 105 (8.9%) out of 1,187 ticks examined (range from 13.3% to 5.6%). Sequencing showed Rickettsia helvetica exclusively. In about 2.6% of Rickettsia-positive ticks, double infection with Anaplasma phagocytophilum was found. Babesia sp. was detected in n= 31 (1.0%) out of 3,113 ticks examined, which originated from 4 different areas. By sequencing, n = 28 (90.0%) were identified as Babesia divergens. Three of all Babesia-positive ticks were identified as harboring Babesia microti. The detection of Anaplasma phagocytophilum, Rickettsia sp. and Babesia sp. demonstrates their possible role as a source of human infection in Germany.

Acquisition of different isolates of Anaplasma phagocytophilum by Ixodes scapularis from a model animal.

The prevalence of etiologic agents in ticks reflects the intensity of their transmission in natural cycles and is an important measure of their potential to cause human disease. The distribution of Anaplasma phagocytophilum within the range of its primary vectors is patchy. Even nearby sites differ dramatically in the prevalence of Anaplasma in questing ticks. We hypothesized that this irregular distribution may be due in part to variations in acquisition rates of different isolates of A. phagocytophilum by I. scapularis ticks from infected animals. BALB/c mice were infected with seven isolates of A. phagocytophilum from different geographical regions: four isolates from the Northeastern United States (Bridgeport, Dawson, Gaillard, NY-8), two from the Midwest (Webster, Sp-Is), and one from California (MRK). Larval ticks were placed on infected mice for 16 consecutive weeks and allowed to feed to repletion. The prevalence of infection in the freshly molted nymphs was then determined by PCR. The proportion of ticks that became infected with either isolate fluctuated over the duration of infection. Mice harboring the isolate Sp-Is were most infectious for ticks at 3 weeks postinfection. Mice infected with the other six isolates exhibited several peaks of infectivity. Timing and relative heights of these peaks differed between isolates. Geographical proximity of the studied isolates did not predetermine their similarity, and isolates originating from the same region differed more in their ability to infect ticks than isolates from different regions. However, it appears unlikely that described differences in agent acquisition by ticks alone are sufficient to account for the irregular distribution of A. phagocytophilum in vector populations.

Detection of four Borrelia burgdorferi genospecies and first report of human granulocytic ehrlichiosis agent in Ixodes ricinus ticks collected in central Italy.

The presence of Borrelia burgdorferi s.l. and of Ehrlichia phagocytophila group was sought by PCR in Ixodes ricinus collected in a protected area of central Italy. Nymphs (n = 1475, gathered in 295 pools of 5 nymphs each) and adult ticks (n = 28) were examined. B. burgdorferi s.l. was detected in 13.8% of the nymph pools; of these, 63.4% were infected by B. valaisiana, 26.8% by B. afzelii, 7.3% by B. garinii, and 2.5% by B. burgdorferi s.s. Only a single adult male tick proved to host B. afzelii. The agent of human granulocytic ehrlichiosis (HGE) was detected in 2.7% of the nymph pools. Two HGE agent-positive nymph pools were also found to be positive for B. garinii and for B. afzelii, respectively. This is the first report from central Italy of the finding of the HGE agent in ticks.

Lyme disease: a search for a causative agent in ticks in south-eastern Australia.

Attempts were made to identify the causative organism of Lyme disease in Australia from possible tick vectors. Ticks were collected in coastal areas of New South Wales, Australia, from localities associated with putative human infections. The ticks were dissected; a portion of the gut contents was examined for spirochaetes by microscopy, the remaining portion inoculated into culture media. The detection of spirochaetes in culture was performed using microscopy, and immunochemical and molecular (PCR) techniques. Additionally, whole ticks were tested with PCR for spirochaetes. From 1990 to 1992, approximately 12,000 ticks were processed for spirochaetes. No evidence of Borrelia burgdorferi or any other spirochaete was recovered from or detected in likely tick vectors. Some spirochaete-like objects detected in the cultures were shown to be artifacts, probably aggregates of bacterial flagellae. There is no definitive evidence for the existence in Australia of B. burgdorferi the causative agent of true Lyme disease, or for any other tick-borne spirochaete that may be responsible for a local syndrome being reported as Lyme disease.

Use of partially engorged female ticks as laboratory animals in microbiological research

Partially engorged female ticks were used as laboratory animals in microbiological research. The ticks, which were inoculated intracoelomally, became a convenient substrate for the detection of viruses, rickettsiae and protozoal parasites. This research concerned the isolation of newly recovered micro-organisms, the study of development, structure and distribution of microbial agents in ticks, and the study of their interaction with other pathogens or symbionts during mixed infection in a tick body. The isolation and maintenance of Rickettsiella phytoseiuli, the organism not of tick-borne origin, was achieved. For use in Central Europe the tick Dermacentor reticulatus is recommended for the above investigations.

Rickettsia rickettsii and Rickettsia montana from Ixodid Ticks in Connecticut

Dermacentor variabilis, infected with spotted fever group rickettsiae, parasitized 8 of 70 raccoons captured in Newtown, Connecticut. The spotted fever agent, Rickettsia rickettsii, was isolated and identified from 4 adult D. variabilis and from 1 nymphal Ixodes texanus removed from raccoons. This verifies the presence of this etiologic agent in ticks in an area where 6 people had clinical signs and symptoms of Rocky Mountain spotted fever (RMSF) and antibodies to R. rickettsii. These are the first isolations of R. rickettsii from D. variabilis in southern New England and the first identified rickettsiae from I. texanus. No rickettsiae were isolated from Ixodes muris or I. cookei. Rickettsia montana was recovered in Vero cell culture from a D. variabilis collected in East Haddam, Connecticut where RMSF is not known to be prevalent.

Frozen sections of arthropods for histological studies and fluorescent antibody investigations

A method for the rapid sectioning of arthropods is described whereby specimens are quick-frozen in gelatin and sectioned in a freezing cabinet at −15 °C. Sections in which the internal anatomical structures were excellently preserved could be prepared regularly from the nymphal and adult ticks, Dermacentor andersoni, Ornithodoros moubata and O. parkeri, and from the cone-nosed bug, Rhodnius prolixus. The usefulness of this frozen section technique was demonstrated or suggested for, (a) histological studies on the relationship of organ systems, (b) localization of parasites and disease agents by means of conventional stains, and (c) fluorescent antibody studies on the development and behavior of disease agents in ticks and other arthropods. By employing conventional stains Trypanosoma rangeli has been localized in the salivary glands of the arthropod host and by fluorescent antibody methods Rickettsia rickettsii was demonstrated successfully in frozen sections prepared from whole ticks.