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Purpose: Flea-borne rickettsioses, collectively referred to as a term for etiological agents Rickettsia felis, Rickettsia typhi, and RFLOs (R. felis-like organisms), has become a public health concern around the world, specifically in the United States. Due to a shared arthropod vector (the cat flea) and clinical signs, discriminating between Rickettsia species has proven difficult. While the effects of microbial coinfections in the vector can result in antagonistic or synergistic interrelationships, subsequently altering potential human exposure and disease, the impact of bacterial interactions within flea populations remains poorly defined. Methods: In this study, in vitro and in vivo systems were utilized to assess rickettsial interactions in arthropods. Results: Coinfection of both R. felis and R. typhi within a tick-derived cell line indicated that the two species could infect the same cell, but distinct growth kinetics led to reduced R. felis growth over time, regardless of infection order. Sequential flea coinfections revealed the vector could acquire both Rickettsia spp. and sustain coinfection for up to 2 weeks, but rickettsial loads in coinfected fleas and feces were altered during coinfection. Conclusion: Altered rickettsial loads during coinfection suggest R. felis and R. typhi interactions may enhance the transmission potential of either agent. Thus, this study provides a functional foundation to disentangle transmission events propelled by complex interspecies relationships during vector coinfections. [ABSTRACT FROM AUTHOR]

Background: In urban Thailand, arboviral infections dominate diagnoses of acute undifferentiated fevers (AUFs) owing to their well-defined epidemiology and characteristic clinical presentations. However, rickettsial diseases, also endemic in this setting, remain under-recognized owing to challenges in early detection. Objective: This study aimed to identify potential rickettsial infections among patients with AUF in Bangkok and vicinity utilizing leftover nucleic acid extracted from serum samples from patients initially suspected of but negative for arbovirus infections. Materials and Methods: A total of 609 nucleic acid samples were screened for rickettsial bacteria using real-time PCR, targeting the 17-kDa common antigen gene of Rickettsia spp. and the 47-kDa gene of Orientia tsutsugamushi. Results: Nine samples were positive for Rickettsia spp. and two were positive for O. tsutsugamushi. DNA sequence and phylogenetic analyses based on partial 17-kDa antigen and citrate synthase (gltA) genes identified the Rickettsia-positive samples as R. typhi in eight cases and R. felis in one case. Analysis of the 56-kDa type-specific antigen gene identified the two O. tsutsugamushi isolates as Gilliam-related genotypes. Although rickettsial diseases typically present with mild symptoms, two patients with R. typhi infection (murine typhus) developed respiratory distress syndrome, highlighting the potential for rare but serious complications. Conclusion: This study underscores the critical importance of differential diagnosis and prompt, effective intervention to prevent complications in suspected cases. [ABSTRACT FROM AUTHOR]

Rickettsia spp. has been detected in dog fleas in Bangkok, Thailand. With the intent of collecting evidence to confirm the presence of rickettsioses in dogs and to assess the level of associated potential for accidental human infection, human buffy coat from patients with fever of unknown origin (n = 168), whole blood samples from dogs (n = 353), and 19 flea groups from our dog sample population were studied during the 2012 to 2014 study period. The presence of Rickettsia was investigated by molecular detection of 23S rRNA gene of Rickettsia genus, citrate synthase (gltA) gene, and 17-kDa outer membrane gene. All positive samples were confirmed by DNA sequence analysis. Using phylogenetic analysis, three groups of Rickettsia were detected, as follows: Rickettsia felis in 8 patients and 8 dogs; R. felis-like sp. in 2 patients, 5 dogs, and 11 flea samples; and Rickettsia typhi in 3 patients. In addition to confirming the presence of R. felis in Thai patients, the findings of this study suggest that R. felis-like sp. isolated from fleas that were symbiotically coexisting with dogs that we evaluated in this study can transmit and cause disease in dogs and humans in Bangkok. [ABSTRACT FROM AUTHOR]

Rickettsia typhi and Rickettsia felis are flea-borne pathogens, which cause murine typhus and flea-borne spotted fever, respectively. Recently, two other flea-borne rickettsiae (phylogenetically similar to R. felis) have been discovered—Rickettsia asembonensis and Candidatus Rickettsia senegalensis. Currently, species-specific identification of detected organisms requires sequencing- or probe-based PCR assays. Our aim was to develop an efficient and inexpensive method to differentiate R. felis and R. felis-like organisms through restriction fragment length polymorphism (RFLP) analysis. Outer membrane protein B sequences of the aforementioned flea-borne rickettsiae were analyzed using DNASTAR Lasergene Core software to focus on the region amplified by the primers 120.2788 and 120.3599. Restriction enzyme digestion sites were identified, and in silico digestions of each species were compared through simulated agarose gels. The enzyme NlaIV was determined to be the most effective at creating a unique banding pattern within the area of interest. To confirm the predicted performance of NlaIV digestion, we tested the DNA of known PCR positive Ctenocephalides felis fleas collected from cats and opossums within Galveston, Texas. DNA from these fleas was amplified using the sca5 primer set 120.2788 and 120.3599. The PCR products were then digested with NlaIV, subjected to polyacrylamide gel electrophoresis, and visualized through ethidium bromide staining. The banding patterns were then compared with the computer-generated digestion patterns. All samples demonstrated a banding pattern consistent with the predicted pattern for the known species, as confirmed by previous sequencing. This RFLP assay was developed to be an efficient and cost-effective method to screen samples for R. felis, R. asembonensis, and Candidatus R. senegalensis. We believe this assay can aid in the epidemiological and ecological studies of flea-borne rickettsiae. [ABSTRACT FROM AUTHOR]

Rickettsia typhi, transmitted by rat fleas, causes most human flea-borne rickettsioses worldwide. Another rickettsia, Rickettsia felis, found in cat fleas, Ctenocephalides felis, has also been implicated as a potential human pathogen. In the continental United States, human cases of flea-borne rickettsioses are reported primarily from the southern regions of Texas and California where the cat flea is considered the principal vector. In California, more than 90% of locally acquired human cases are reported from suburban communities within Los Angeles and Orange counties despite the almost ubiquitous presence of cat fleas and their hosts throughout the state. The objective of this study is to assess the presence and infection rate of Rickettsia species in cat fleas from selected endemic and nonendemic regions of California. Cat fleas were collected from cats in Los Angeles County (endemic region) and Sacramento and Contra Costa counties (nonendemic region). Sequencing of 17 amplicons confirmed the presence of R. felis in both the endemic and non-endemic regions with a calculated maximum likelihood estimation of 131 and 234 per 1000 fleas, respectively. R. typhi was not detected in any flea pools. Two R. felis-like genotypes were also detected in fleas from Los Angeles County; Genotype 1 was detected in 1 flea pool and Genotype 2 was found in 10 flea pools. Genotype 1 was also detected in a single flea pool from Sacramento County. Results from this study show that R. felis is widespread in cat flea populations in both flea-borne rickettsioses endemic and nonendemic regions of California, suggesting that a high prevalence of this bacterium in cat fleas does not predispose to increased risk of human infection. Further studies are needed to elucidate the role of R. felis and the two R. felis-like organisms as etiologic agents of human flea-borne rickettsioses in California. [ABSTRACT FROM AUTHOR]

Small Indian mongooses (SIMs, Herpestes auropunctatus) have invasively inhabited over 60 islands worldwide. They have been confirmed as a reservoir of rabies, leptospirosis, and salmonellosis; however, their role in the epidemiology of other zoonoses is little known. On St. Kitts, as well as other islands, SIMs harbor Ctenocephalides felis, which can vector several zoonotic diseases. In this study, SIMs were examined for fleas, and the collected fleas analyzed by PCR and DNA sequencing for Bartonella henselae, Rickettsia felis, Yersinia pestis, and Dipylidium caninum. Of the 87 SIMs, 75 (86.2%) harbored C. felis. C. felis recovered from nine (10.3%), one (1.1%), and one (1.1%) of the SIMs was positive for B. henselae, R. felis, and D. caninum, respectively. These data indicate that SIMs serve as an additional reservoir of B. henselae and R. felis, which should be taken into consideration in control and prevention of these rapidly emerging zoonoses. [ABSTRACT FROM AUTHOR]

High prevalence of Rickettsia felis in patients with fever of unknown origin was revealed in the north-central Bangladesh from 2012 to 2013. Subsequently, in this study, prevalence of R. felis in cats and cat fleas ( Ctenocephalides felis), together with febrile patients, was studied by PCR detection of 17 kDa antigen gene and DNA sequencing. R. felis was detected in 28% (28/100) and 21% (14/68) of cat blood and cat flea samples, respectively, whereas 42% (21/50) of patients were positive for R. felis. R. felis-positive cat fleas were detected at significantly higher rate on R. felis-positive cats. The results suggested a potential role of cats and cat fleas for transmission of R. felis to humans in Bangladesh. [ABSTRACT FROM AUTHOR]

To add to the limited information on Rickettsia in mosquitoes in China, we carried out a PCR survey on convenience samples of 3051 mosquitoes collected with hand nets in and around domestic dwellings in 25 provinces. Five species of mosquitoes were identified: Culex pipiens pallens (n = 1620), Aedes albopictus (806), Armigeres subalbatus (377), Anopheles sinensis (168), and Culex tritaeniorhynchus (80). A Rickettsia nested-PCR targeting the variable domain of gltA showed Rickettsia felis in four mosquito species of 16 provinces Cx. pipiens pallens (1.8%, 29/1620); Ae. albopictus (1.2%, 10/806); An. sinensis (1.2%, 2/168); and Ar. subalbatus (2.1%, 8/377). Rickettsia bellii was also widespread, occurring in 12 provinces and 2 species: Cx. pipiens pallens (4.3%, 69/1620) and An. sinensis (0.6%, 1/168). R. felis and R. bellii were found in almost similar numbers in female [1.5% (27/1809) and 1.2% (21/1809), respectively] as in male mosquitoes [1.8% (22/1242) and 4.0% (49/1242), respectively]. Our results indicated that mosquitoes in China are widely infected with R. felis, the agent of human flea-borne spotted fever, and that R. bellii can also occur outside of the Americas and its usual tick hosts. [ABSTRACT FROM AUTHOR]

Little is known of the endemicity of flea-borne rickettsiae in Kazakhstan. Thus, a survey for rickettsiae within great gerbil fleas was conducted in Almaty oblast. High prevalence of Rickettsia asembonensis was detected among Xenopsylla gerbilli, demonstrating that flea-borne rickettsiae are endemic to southeastern Kazakhstan. Interestingly, no Rickettsia typhi were detected in these same fleas. [ABSTRACT FROM AUTHOR]

To expand the documentation of rickettsioses in Indonesia, we conducted an ectoparasite and small mammal investigation involving four major islands: Java, Sumatra, Sulawesi, and Kalimantan. Coastal and highland regions on each island surveyed were chosen to represent different ecologies in Indonesia. Indication of the presence of Rickettsia spp. was evident in all areas sampled. Typhus group rickettsiae-specific antibodies had significantly higher prevalence among small mammals captured in Java compared to the other islands surveyed (78% in coastal and 50% in highland regions) and the prevalence of spotted fever group rickettsiae-specific antibodies was significantly higher in Kalimantan than the other islands investigated. Hosts and vectors were restricted by Rickettsia spp. but not by coastal or highland regions. Our findings expand the range in which rickettsial pathogens have been documented within the Indonesian archipelago and point to a significant risk to human health. [ABSTRACT FROM AUTHOR]

This survey is aimed at investigation of species composition of fleas removed from small terrestrial mammals captured in rural, suburban, and urban types of habitat and molecular screening of the presence and diversity of Rickettsia species in collected ectoparasites. In total, 279 fleas (Siphonaptera) belonging to 9 species of 2 families, Ceratophyllidae and Hystrichopsyllidae, were collected from 115 (46%) out of 250 trapped small mammals of eight species (Apodemus agrarius, Apodemus flavicollis, Apodemus uralensis, Myodes glareolus, Microtus arvalis, Microtus subterraneus, Crocidura leucodon, and Sorex minutus). Rickettsia spp. were found in 2.5% (7/279) of tested fleas, namely in Ctenophthalmus agyrtes, Ctenophthalmus solutus, Ctenophthalmus uncinatus, Megabothris turbidus, and Amalareus penicilliger. Rickettsia felis, Rickettsia helvetica, and unidentified Rickettsia species were detected in fleas infesting small mammals in Eastern Slovakia. The results of the study suggest that some species of rickettsiae have a different range of arthropod vectors. [ABSTRACT FROM AUTHOR]

Members of the order Rickettsiales are small, obligate intracellular bacteria that are vector-borne and can cause mild to fatal diseases in humans worldwide. There is little information on the zoonotic rickettsial pathogens that may be harbored by dogs from rural localities in South Africa. To characterize rickettsial pathogens infecting dogs, we screened 141 blood samples, 103 ticks, and 43 fleas collected from domestic dogs in Bushbuckridge Municipality, Mpumalanga Province of South Africa, between October 2011 and May 2012 using the reverse line blot (RLB) and Rickettsia genus and species-specific quantitative real-time PCR (qPCR) assays. Results from RLB showed that 49% of blood samples and 30% of tick pools were positive for the genus-specific probes for Ehrlichia/ Anaplasma; 16% of the blood samples were positive for Ehrlichia canis. Hemoparasite DNA could not be detected in 36% of blood samples and 30% of tick pools screened. Seven (70%) tick pools and both flea pools were positive for Rickettsia spp; three (30%) tick pools were positive for Rickettsia africae; and both flea pools (100%) were positive for Rickettsia felis. Sequencing confirmed infection with R. africae and Candidatus Rickettsia asemboensis; an R. felis-like organism from one of the R. felis-positive flea pools. Anaplasma sp. South Africa dog strain (closely related to Anaplasma phagocytophilum), A. phagocytophilum, and an Orientia tsutsugamushi-like sequence were identified from blood samples. The detection of emerging zoonotic agents from domestic dogs and their ectoparasites in a rural community in South Africa highlights the potential risk of human infection that may occur with these pathogens. [ABSTRACT FROM AUTHOR]

Background: In Lyme-endemic areas, limited research has investigated the risk of tick-borne diseases among frontline workers in fire management. This study aimed to compare the exposure histories to ticks and tick-borne pathogens, knowledge, and prevention practices between individuals engaged in fire-related forest management and those participating in recreational activities within Santa Cruz County, CA, in an area of high risk of tick exposure. Methods: Blood samples from 55 forest workers and 58 members of the public were tested for bacterial DNA of and antibodies to Borrelia burgdorferi, Anaplasma phagocytophilum, and Rickettsia species. In addition, a questionnaire was administered to 54 workers and 84 members of the public to identify gaps in knowledge and prevention practices. Results: Although workers had a higher percentage of positive B. burgdorferi antibodies than the public (5.5% compared with 1.7%), the difference was not statistically significant. Conversely, rickettsial antibodies were more prevalent among the public (17.2% versus 3.6% for workers), but specificity to pathogenic bacteria could not be confirmed. No DNA for the three pathogens or antibodies against A. phagocytophilum were detected. Many workers and members of the public reported tick bites (67.4% of participants), with a notable 11% increase among workers in the odds of being bitten for each additional year spent working in forests. Although workers took greater precautions, significant knowledge and practice gaps were identified among both populations, such as an inability to distinguish tick species from common arthropods (mites, spiders, fleas), overestimating the size of ticks, and inappropriate tick-removal techniques. Conclusion: This study underscores the risk of tick-borne diseases faced by fire management workers in Lyme disease–endemic regions. The findings emphasize the necessity for future studies of Lyme disease within this population, and highlight the urgent need for enhanced training programs to minimize these risks. [ABSTRACT FROM AUTHOR]

Cats and cat fleas (Ctenocephalides felis) are vectors of the zoonotic bacterial pathogens Bartonella henselae and Rickettsia felis, which are the causative agents of “cat scratch disease” and “cat flea typhus,” respectively. In the surroundings of Vienna (Austria), we identified 11 (10.5%; n = 105) B. henselae-positive fleas originating from 8 cats (20.5%; n = 39). One flea was positive for R. felis. There should be high levels of awareness among veterinarians and animal keepers as to the handling of cats, especially if free roaming, stray, or feral. [ABSTRACT FROM AUTHOR]

Murine typhus is a flea-borne typhus group rickettsiosis caused by Rickettsia typhi. Once a prevalent disease in the United States, the use of dichlorodiphenyltrichloroethane in the 1940s broke the classic rat-rat flea cycle of transmission, and the remaining endemic foci are now believed to be associated with opossums and the cat flea (Ctenocephalides felis). In Galveston, Texas murine typhus has re-emerged as a cause of febrile illness, and 7% of fleas collected from opossums are infected with R. typhi. In this study, we sought to explore the prevalence of rickettsiae associated with fleas on cats, as these animals have been speculated to play a role in the epidemiology of murine typhus. Fleas were collected from feral cats entering a local veterinary clinic as part of a trap, spay, neuter, and release program. Fleas were identified and subjected to analysis by PCR and sequencing. An estimation of the minimum infection rate (MIR) of pooled samples was performed. Three hundred fourteen fleas (all C. felis) were collected from 24 cats. Sequences for the outer membrane protein B gene revealed R. typhi in one pool (MIR 0.3%), Rickettsia felis in four pools (MIR 1.3%), Rickettsia asembonensis in one pool (MIR 0.3%), and "Candidatus R. senegalensis" in six pools (MIR 2.0%). Results were confirmed by sequencing portions of the rickettsial citrate synthase and 17-kD protein gene. In this study, the presence of R. typhi in fleas from cats suggests that in Galveston, there exists a small but measurable risk to humans who come into contact with flea-infested cats. Despite this, we believe that the low prevalence from cat-collected fleas, compared with that previously detected from opossums, makes cats less likely to play a role in the maintenance of R. typhi in this region. The significance of other identified flea-borne rickettsiae is yet to be elucidated. [ABSTRACT FROM AUTHOR]

The aim of this work was to detect and identify Rickettsia species in ticks collected in rural areas of Villeta, Colombia. Tick specimens were collected from domestic animals and walls of houses in five rural villages of Villeta town and from humans in Naranjal village (same town). Moreover, a flea collected from the same area was also processed. DNA was extracted and tested by conventional, semi-nested, and nested PCR reactions targeting rickettsial genes. In the ticks collected from humans from Naranjal village, a nymph of Amblyomma cajennense sensu lato was amplified using primers for ompA and sequenced (100% identity with ' Candidatus Rickettsia amblyommii'). Last, three amplicons from the Ctenocephalides felis flea, corresponding to gltA, ompB, and 16S rRNA genes, showed high identity with R. felis (98.5%, 97.3%, and 99.2%, respectively) and ' Candidatus Rickettsia asemboensis' (99.7% and 100%, respectively). To our knowledge, these results correspond to the first molecular detection in Colombia of ' Candidatus Rickettsia amblyommii' and ' Ca. Rickettsia asemboensis' in fleas. [ABSTRACT FROM AUTHOR]

Despite several reports worldwide documenting the presence of Rickettsia asembonensis in samples derived from ectoparasites, animals and more recently humans, genomic information of these specimens remains scarce, and when available, is usually limited to small genomic fragments of limited value. We generated complete sequences for two conserved (17-kDa antigen gene and gltA) and three variable (sca4, ompB and ompA) genes in five R. asembonensis DNA samples detected in cat and dog fleas in Peru. Complete gene sequences were used to conduct multi-locus sequence typing and phylogenetic analyses to assess diversity and infer relationships among strains and other reference sequences. The 17-kDa antigen gene was highly conserved across Rickettsia species. Of the variable genes ompB was the most variable, but this diversity was not captured through phylogenetics alone even when efforts were made to maximize potential diversity in terms of flea species, animal host and location. Through a combination of de novo and reference-based genome assembly we identified a 75 bp insertion in ompA that encodes a 25 aa repetitive motif found in other Rickettsia species, but not present in the original prototype strain from Kenya. R. asembonensis has only recently been shown to be a bona-fide human pathogen. As such, and compounded by a lack of available genomic information, it remains understudied. Our work directly addresses the lack of genomic information available worldwide for the study of these novel Rickettsia species and specifically contributes to our understanding of the diversity and molecular epidemiology of R. asembonensis in Peru. [ABSTRACT FROM AUTHOR]

Fleas are carriers for many largely understudied zoonotic, endemic, emerging, and re-emerging infectious disease agents, but little is known about their prevalence and role as a vector in Africa. The aim of this study was to determine the diversity of fleas and the prevalence of infectious agents in them collected from human dwellings in western Kenya. A total of 306 fleas were collected using light traps from 33 human dwellings; 170 (55.56%) were identified as Ctenocephalides spp., 121 (39.54%) as Echidnophaga gallinacea, 13 (4.25%) as Pulex irritans, and 2 (0.65%) as Xenopsylla cheopis. Of the 306 individual fleas tested, 168 (54.9%) tested positive for rickettsial DNA by a genus-specific quantitative real-time PCR (qPCR) assay based on the 17-kDa antigen gene. Species-specific qPCR assays and sequencing revealed presence of Rickettsia asembonensis in 166 (54.2%) and Rickettsia felis in 2 (0.7%) fleas. Borrelia burgdorferi, normally known to be carried by ticks, was detected in four (1.3%) flea DNA preparations. We found no evidence of Yersinia pestis, Bartonella spp., or Orientia spp. Not only were Ctenocephalides spp. the most predominant flea species in the human dwellings, but also almost all of them were harboring R. asembonensis. [ABSTRACT FROM AUTHOR]

Rickettsia species cause rickettsioses, which are zoonotic diseases found worldwide, and are transmitted by arthropods such as lice, fleas, ticks, and mites. In Thailand, flea infestations are common among cats and dogs. This study aimed at determining the exposure to spotted fever group rickettsiae (SFGR) of cats in surrounding areas of Rajabhat Maha Sarakham University, Muang district, Maha Sarakham province and rickettsial infection among cat fleas, Ctenocephalides felis, collected from dogs of the surrounding area of Waeng Noi district, Khon Kaen province. Forty-two cat sera were assessed for IgG antibody titers against SFGR by a group-specific enzyme-linked immunosorbent assay. The prevalence of seroreactive cats was 4.76% (2/42). DNA preparations from 23 individual cat fleas from three dogs were assessed by Rickettsia genus-specific, group-specific, and species-specific quantitative real-time PCR (qPCR) assays. Positive results were confirmed by ompB gene fragment sequencing. Twenty-one of 23 cat fleas were positive for Rickettsia asembonensis, and the other two DNA preparations were negative for rickettsial DNA. This study's finding indicates that companion cats and dogs in Northeast Thailand are exposed to SFGR and that exposure may be due to infection with R. asembonensis, an organism known to infect humans, monkeys, and dogs. Clinicians for humans and animals in Northeast Thailand should be aware of rickettsial infections among their patients. [ABSTRACT FROM AUTHOR]

In a sanctuary located on the island of Malta, 23 clinically healthy cats randomly selected were sampled for blood and fleas. Only fleas were collected from 35 cats. All fleas were identified as Ctenocephalides felis, except for one specimen of Ctenocephalides canis. To the best of the authors' knowledge, this may be the first time to establish the occurrence of Bartonella koehlerae and B. clarridgeiae, as well as of Candidatus Mycoplasma haemominutum in the blood samples of 11 cats (47.82% [95% CI: 29.33–67.04]) with conventional PCR assays. One or more pathogens were found in 54 (96.42% [95% CI: 86.74–99.70]) out of 56 pooled flea samples, the most prevalent was Rickettsia felis. The DNA of Bartonella henselae, the commonest etiological agent of cat scratch disease, was detected first time in a pooled flea sample of a cat. [ABSTRACT FROM AUTHOR]

Bacterial arthropod-borne pathogens can often cause fever in Africa, but rural laboratories in these settings are usually too basic to provide a precise picture of their epidemiological impact. Our aim was to determine the prevalence of bacterial pathogens in fleas and lice in a rural area of southeast Ethiopia. Between July and November 2013, we extracted DNA from 91 fleas (Ctenocephalides felis [n = 50; 54.9%], Pulex irritans [n = 37; 40.1%], and C. canis [n = 4; 4.4%] and 30 lice (Pediculus humanus capitis [n = 16; 53.3%] and Pediculus humanus humanus [n = 14; 46.7%]), using two quantitative PCR (qPCR) analyses to look for bacteria from the genera: Anaplasma, Bartonella, Borrelia, Coxiella, Ehrlichia, Francisella, and Rickettsia. Of the 91 fleas analyzed, pathogens were present in 79 (86.8%), including Rickettsia felis (n = 41; 45%), Anaplasma platys (n = 40; 44.0%), Rickettsia monacensis (n = 2; 2.2%), Ehrlichia muris-like agent (n = 1; 1.1%), and Bartonella clarridgeiae (n = 1; 1.1%). P. irritans was the flea species most frequently infected with A. platys (67.7%), followed by C. felis (30.7%) (p < 0.001). Of the 30 lice identified, pathogens were present in 7 (23.3%): Bartonella quintana (n = 4; 16.7%), E. muris (n = 2, 6.7%), and Borrelia recurrentis (n = 1, 3.3%). Thus, in this rural area of Africa, fleas and lice can transmit parasitic pathogens to humans, causing febrile symptoms. [ABSTRACT FROM AUTHOR]

Human infections with Rickettsia felis have been reported worldwide. Recent studies have revealed the presence of many closely related but unique rickettsiae, referred to as Rickettsia felis-like organisms (RFLO), identified in various arthropods. Due to the recent discovery of the lack of specificity of earlier R. felis-specific assays, there has become a need to develop a new generation of R. felis-specific molecular assays that will differentiate R. felis not only from other rickettsiae but more importantly from other members of the R. felis genogroup that may not be pathogenic to humans. This new generation of assays is essential for determining the true risk for flea-borne spotted fever (FBSF) by surveying arthropod vectors/hosts. Because of the lack of specificity of previous assays developed to detect R. felis infections, prior surveys may have overestimated the prevalence of R. felis in arthropod vectors and thus the perceived risk of FBSF. We have developed a specific quantitative real-time polymerase chain reaction (qPCR) assay to detect R. felis (RfelB). Specificity of the assay was determined by testing it with a panel of 17 related Rickettsia species and 12 nonrickettsial bacterial DNA preparations. The RfelB qPCR assay was positive for R. felis DNA and negative for all of the 17 related Rickettsia species and 12 nonrickettsia bacterial DNA preparations. The limit of detection of the RfelB qPCR assay was determined to be two copies (two genoequivalents) per microliter of R. felis target ompB fragment-containing plasmid. Validation of the RfelB qPCR assay was accomplished by testing 83 previously sequence-confirmed R. felis and RFLOs containing DNA preparations from human and flea samples collected from different geographical locations around the world. This assay will be useful for rapid detection, identification, and enumeration of R. felis, an emerging human pathogen of worldwide importance, from both clinical and environmental samples. [ABSTRACT FROM AUTHOR]

Rickettsia felis is an obligate, intracellular, Gram-negative bacterium and a member of the transitional group rickettsiae. This bacterium has been shown to grow in vitro in amphibian, tick, and mosquito cell lines. Here, we present data to show the growth of R. felis strain LSU in Drosophila melanogaster S2 cells, an embryonic, hemocytic cell line with phagocytic properties. R. felis LSU was isolated from Ixodes scapularis E6 (ISE6) cells and used to infect S2 cells, grown at 25°C. By 19 days postinfection, the S2 cells were 100% infected with R. felis as determined by Acridine Orange and Diff-Quik staining. A species-specific R. felis qPCR assay was used to demonstrate that the kinetics associated with the S2 cell culture infection involved a lag/adaptation phase, followed by continued growth to 20 days postinfection. Moreover, R. felis organisms were observed in the S2 cells using transmission electron microscopy and a polyclonal antibody against spotted fever rickettsiae. The ability to use D. melanogaster S2 cells for growing rickettsial agents is a useful tool owing to the ease of manipulation of the S2 cultures and the wide-ranging possibility of Drosophila resources available for future studies. [ABSTRACT FROM AUTHOR]

The flea-borne rickettsioses murine typhus (Rickettsia typhi) and flea-borne spotted fever (FBSF) (Rickettsia felis) are febrile diseases distributed among humans worldwide. Murine typhus has been known to be endemic to Kenya since the 1950s, but FBSF was only recently documented in northeastern (2010) and western (2012) Kenya. To characterize the potential exposure of humans in Kenya to flea-borne rickettsioses, a total of 330 fleas (134 pools) including 5 species (Xenopsylla cheopis, Ctenocephalides felis, Ctenocephalides canis, Pulex irritans, and Echidnophaga gallinacea) were collected from domestic and peridomestic animals and from human dwellings within Asembo, western Kenya. DNA was extracted from the 134 pooled flea samples and 89 (66.4%) pools tested positively for rickettsial DNA by 2 genus-specific quantitative real-time PCR (qPCR) assays based upon the citrate synthase (gltA) and 17-kD antigen genes and the Rfelis qPCR assay. Sequences from the 17-kD antigen gene, the outer membrane protein (omp)B, and 2 R. felis plasmid genes (pRF and pRFd) of 12 selected rickettsiapositive samples revealed a unique Rickettsia sp. (n = 11) and R. felis (n = 1). Depiction of the new rickettsia by multilocus sequence typing (MLST) targeting the 16S rRNA (rrs), 17-kD antigen gene, gltA, ompA, ompB, and surface cell antigen 4 (sca4), shows that it is most closely related to R. felis but genetically dissimilar enough to be considered a separate species provisionally named Candidatus Rickettsia asemboensis. Subsequently, 81 of the 134 (60.4%) flea pools tested positively for Candidatus Rickettsia asemboensis by a newly developed agentspecific qPCR assay, Rasemb. R. felis was identified in 9 of the 134 (6.7%) flea pools, and R. typhi the causative agent of murine typhus was not detected in any of 78 rickettsia-positive pools assessed using a species-specific qPCR assay, Rtyph. Two pools were found to contain both R. felis and Candidatus Rickettsia asemboensis DNA and 1 pool contained an agent, which is potentially new. [ABSTRACT FROM AUTHOR]

Introduction: Rickettsia felis produces a syndrome indistinguishable from murine typhus, which has been described in Spain. R. felis is transmitted to humans by fleas. Although no clinical case has been described so far, serologic evidence of infections in humans, cats, and dogs has been obtained in our area. However, no study has been conducted regarding its presence in vectors. Recognition of routes of transmission is of great importance to prevent infection in humans. Taking into account these results, R. felis seems to be present in animals that are in contact with humans. The aim of this study was to determine the presence of R. felis in the fleas of cats and dogs from Northeast Spain, to show the presence of peridomestic cycle in our area. Materials and Methods: Between May 2006 and July 2008, 78 fleas were collected. Sixty-three fleas were recovered from kennels. Most of them were collected from cages and a few of them on dogs and cats living in kennels. Fifteen fleas were collected from dogs and cats attended at a veterinary clinic. Fleas were rinsed with ethanol, dried, identified, and stored at 4°C. DNA was extracted from each flea individually. Rickettsial DNA was determined by quantitative real-time polymerase chain reaction. OmpB-specific primers and molecular beacon probes targeting specifically R. felis were used. Results: All 78 fleas were identified as Ctenocephalides felis. R. felis was detected in 34 (43.6%) fleas. No nucleic acids were amplified from negative controls and expected results were obtained from positive controls. Eight positive samples were also confirmed by sequencing. Conclusions: R. felis was found in a high percentage of Ct. felis from cats and dogs. It seems that there is a peridomestic cycle in Northeast Spain, which would allow contact of R. felis with humans. [ABSTRACT FROM AUTHOR]

Flea-borne diseases (FBDs) impact both human and animal health worldwide. Because adult fleas are obligately hematophagous and can harbor potential pathogens, fleas act as ectoparasites of vertebrates, as well as zoonotic disease vectors. Cat fleas (Ctenocephalides felis) are important vectors of two zoonotic bacterial genera listed as priority pathogens by the National Institute of Allergy and Infectious Diseases (NIAID-USA): Bartonella spp. and Rickettsia spp., causative agents of bartonelloses and rickettsioses, respectively. In this study, we introduce the first microbiome analysis of C. felis samples from California, determining the presence and abundance of relevant pathogenic genera by characterizing the cat flea microbiome through 16S rRNA next-generation sequencing (16S-NGS). Samples from both northern (NoCal) and southern (SoCal) California were assessed to expand current knowledge regarding FBDs in the state. We identified Rickettsia and Bartonella, as well as the endosymbiont Wolbachia, as the most abundant genera, followed by less abundant taxa. In comparison to our previous study screening Californian cat fleas for rickettsiae using PCR/digestion/sequencing of the ompB gene, the 16S-NGS approach applied herein showed a 95% level of agreement in detecting Rickettsia spp. There was no overall difference in microbiome diversity between NoCal and SoCal samples. Bacterial taxa identified by 16S-NGS in this study may help to improve epidemiological investigations, pathogen surveillance efforts, and clinical diagnostics of FBDs in California and elsewhere. [ABSTRACT FROM AUTHOR]

Introduction: Rickettsia felis is an emerging agent considered a human threat; although its natural reservoir and agent of transmission is the cat flea, it has been also found in other vectors. R. felis has been identified in Chile in cat fleas and in one specimen of Rhipicephalus sanguineus collected in the Metropolitan Region. The objective of this study was to detect the presence of Rickettsia spp. in R. sanguineus from dogs of two different and distant geographical areas in Chile. Material and Methods: We performed a domiciliary sampling in urban and rural localities of two distant areas of the country--the Metropolitan Region in the center and the northern city of Arica. A total of 460 households were visited; one dog per household was included in the study and ectoparasites were collected from them. Results: R. sanguineus was found in 50% of the 460 dogs. R. felis was identified by amplification and sequencing of gltA, ompA, and ompB genes in R. sanguineus from both regions, with predominance in Arica. Discussion: The presence of R. felis in R. sanguineus from two distant regions of Chile suggests that this rickettsial agent is well established in the country. Considering that no human spotted fever group infections have been recognized in the country, the results should alert clinicians about such possible cases. The role of R. sanguineus in the epidemiology and transmission of R. felis should be further investigated. [ABSTRACT FROM AUTHOR]

Candidatus Rickettsia asemboensis was identified molecularly in fleas collected in 2009 from Asembo, Kenya. Multilocus sequence typing using the 17-kD antigen gene, rrs, gltA, ompA, ompB, and sca4 demonstrated that Candidatus R. asemboensis is closely related to Rickettsia felis but distinct enough to be considered for separate species classification. Following this molecular characterization of Candidatus R. asemboensis, the in vitro cultivation of this bacterium was then performed. We used Ctenocephalides canis and Ctenocephalides felis fleas removed from dogs in Kenya to initiate the in vitro isolation of Candidatus R. asemboensis. Successful cultures were obtained using Drosophila melanogaster S2 and Aedes albopictus C6/36 cell lines. Cytological staining and quantitative real-time PCR (qPCR) assays were used to visualize/confirm the culture of the bacteria in both cell lines. Sequencing of fragments of the 17-kD antigen gene, gltA, and ompB genes confirmed the identity of our Candidatus R. asemboensis isolates. To date, we have passaged Candidatus R. asemboensis 12 times through S2 and C6/36 cells, and active and frozen cultures are currently being maintained. This is the first time that a R. felis-like organism has been grown and maintained in culture and is therefore the first time that one of them, Candidatus R. asemboensis, has been characterized beyond molecular typing. [ABSTRACT FROM AUTHOR]

Surveillance for Rickettsia spp. is urgently needed due to the recent emergence of many novel rickettsioses around the globe, but previous studies in Taiwan have been limited to small areas and no investigation of infections in vertebrate hosts has ever been attempted. We surveyed rickettsial infections systematically in small-mammal hosts trapped between 2006 and 2010 throughout Taiwan. Fragments of ompB and gltA genes in the liver, spleen, and kidney of mammals were targeted by nested polymerase chain reaction. We trapped 1375 individuals of 10 species, among which Rattus losea was the most common (54.6%), followed by Suncus murinus (20.6%) and Mus caroli (10.6%). The overall rate of Rickettsia infections in the liver, spleen, or kidney of 309 assayed small mammals was 60.5%, with a rate of infection ≥50% for each mammal species. DNA nucleotide sequences of 184 successfully sequenced genes were most similar to nine Rickettsia species: Rickettsia conorii, R. felis, R. japonica, R. raoultii, R. rickettsii, Rickettsia sp. IG-1, Rickettsia sp. TwKM01, Rickettsia sp. TwKM02, and R. typhi. Our results suggest that several novel Rickettsia spp. are common and widespread across various habitats throughout Taiwan and suggest the need for further study of emerging rickettsioses in Taiwan. [ABSTRACT FROM AUTHOR]

In the United States, Dermacentor variabilis and Dermacentor andersoni are considered key vectors for Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever. Through regional surveillance, a wide diversity of Rickettsia spp. have been documented in D. variabilis, and Dermacentor spp. has been suggested as potential vectors for various other pathogens, including Babesia spp. and Ehrlichia canis. To better define the prevalence and diversity of pathogens in Dermacentor spp. across the United States, 848 ticks collected from dogs and cats in 44/50 states in 2018–2019 were tested by PCR for Rickettsia spp.-specific 17 kDa and ompA gene fragments; a subset of Dermacentor spp. was also tested with PCR, targeting fragments of the 18S and large subunit region rRNA genes of Babesia spp. and 16S rRNA genes of E. canis. Rickettsia spp. was identified in 12.5% (106/848) of ticks. Species detected include Rickettsia montanensis (n = 64 ticks), Rickettsia bellii (n = 15 ticks), Rickettsia rhipicephali (n = 13 ticks), Rickettsia peacockii (n = 8 ticks), Rickettsia amblyommatis (n = 3 ticks), Rickettsia cooleyi (n = 1 tick), and unclassified Rickettsia spp. (n = 2 ticks). Ticks with R. montanensis and R. bellii were submitted from every U.S. region; R. rhipicephali was predominantly detected in ticks from the southern half of the United States, and all R. peacockii-positive ticks were D. andersoni that originated from the Rocky Mountain states. Ehrlichia canis was not detected in any Dermacentor spp., and Babesia conradae was detected in two Dermacentor albipictus. Because most ticks had fed on dogs or cats before submission, these findings do not implicate a given Dermacentor sp. as a primary vector of these agents, but in regard to Rickettsia spp., the data do support other published work showing D. variabilis harbors a diversity of Rickettsia species with unknown implications for animal and human health. [ABSTRACT FROM AUTHOR]

Background and Objectives: Vector-borne bacterial diseases represent a substantial public health burden and rodents have been recognized as important reservoir hosts for many zoonotic pathogens. This study investigates bacterial pathogens in a small mammal community of the southwestern United States of America. Methods: A total of 473 samples from 13 wild rodent and 1 lagomorph species were tested for pathogens of public health significance: Bartonella, Brucella, Yersinia, Borrelia, Rickettsia spp., and Anaplasma phagocytophilum. Results: Three animals were positive for Yersinia pestis, and one Sylvilagus audubonii had a novel Borrelia sp. of the relapsing fever group. No Brucella, Rickettsia, or A. phagocytophilum infections were detected. Bartonella prevalence ranged between 0% and 87.5% by animal species, with 74.3% in the predominant Neotoma micropus and 78% in the second most abundant N. albigula. The mean duration of Bartonella bacteremia in mark-recaptured N. micropus and N. albigula was 4.4 months, ranging from <1 to 18 months, and differed among Bartonella genogroups. Phylogenetic analysis of the Bartonella citrate synthase gene (gltA) revealed 9 genogroups and 13 subgroups. Seven genogroups clustered with known or previously reported Bartonella species and strains while two were distant enough to represent new Bartonella species. We report, for the first time, the detection of Bartonella alsatica in North America in Sylvilagus audubonii and expand the known host range of Bartonella washoensis to include Otospermophilus variegatus. Interpretation and Conclusion: This work broadens our knowledge of the hosts and geographic range of bacterial pathogens that could guide future surveillance efforts and improves our understanding of the dynamics of Bartonella infection in wild small mammals. [ABSTRACT FROM AUTHOR]

Ixodes ricinus is a potential vector for some of the tick-borne microorganisms that can cause significant diseases in animals and humans. This study aims to determine the prevalence of Anaplasma, Rickettsia, Bartonella, and Francisella species in host-seeking ticks collected from the forest areas in the Sinop region located in the northernmost part of Turkey. Between May and July 2017, a total of 135 tick pools formed from 2571 of the 2734 ticks collected out of the vegetation. Samples of each pool were homogenized and analyzed by PCR. Infection prevalence was statistically analyzed in view of the maximum likelihood estimation (MLE) with a 95% confidence interval (CI). DNA of the infectious agents was determined only in the adult and nymph pools of I. ricinus. MLE values of Anaplasma spp. and Bartonella spp. in 58 pools formed from 517 of I. ricinus adults were 1.20% (95% CI: 0.50–2.49) and 0.80% (95% CI: 0.26–1.91), respectively. In 42 pools generated from 1222 of I. ricinus nymph, MLE values of infection prevalence for Anaplasma spp. and Bartonella spp. were calculated to be 0.17% (95% CI: 0.03–0.54) and 0.34% (95% CI: 0.11–0.82) in respective order. MLE values for Rickettsia spp. were 7.55% (95% CI: 5.21–10.69) and 0.52% (95% CI: 0.22–1.083) for the adult and nymph I. ricinus, respectively. The DNA of Francisella tularensis was not detected in any tick pool. The outcomes of this research are the first molecular evidence of Bartonella spp. and Bartonella henselae in questing I. ricinus in Turkey. The results also suggested that I. ricinus plays considerable roles in enzootic transmission cycles of Anaplasma phagocytophilum, B. henselae, and Rickettsia monacensis in the northernmost region of Turkey. [ABSTRACT FROM AUTHOR]

There is a paucity of data on human exposure to tick-borne pathogens in the western United States. This study reports prevalence of antibodies against three clinically important tick-borne pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, and Rickettsia spp.) among 249 people in five counties in northern California. Individuals from Humboldt County were recruited and answered a questionnaire to assess risk of exposure to tick-borne pathogens. Samples from other counties were obtained from a blood bank and were anonymized. Seventeen (6.8%) samples were seropositive for antibodies against at least one pathogen: five for A. phagocytophilum, eight for B. burgdorferi, and four for Rickettsia spp. Women and people aged 26–35 had higher seroprevalence compared to other demographic groups. Santa Cruz County had no seropositive individuals, northern Central Valley counties had three seropositive individuals (all against A. phagocytophilum), and Humboldt County had 14 (all three pathogens), a significant, four-fold elevated risk of exposure. The Humboldt County questionnaire revealed that a bird feeder in the yard was statistically associated with exposure to ticks, and lifetime number of tick bites was associated with increasing age, time watching wildlife, and time hiking. Three-quarters of respondents were concerned about tick-associated disease, 81.0% reported experiencing tick bites, and 39.0% of those bitten reported a tick-borne disease symptom, including skin lesions (76.4%), muscle aches (49.1%), joint pain (25.5%), or fever (23.6%). Despite high levels of concern, many individuals who had been bitten by a tick were not tested for a tick-borne pathogen, including those with consistent symptoms. We highlight the need for further research and dissemination of information to residents and physicians in Northern California regarding tick-associated disease, so that appropriate medical attention can be rapidly sought and administered. [ABSTRACT FROM AUTHOR]

Bacterial arthropod-borne pathogens are a common cause of fever in Africa, but their precise impact is unknown and usually underdiagnosed in the basic rural laboratories of low-resourced African countries. Our aim was to determine the prevalence of arthropod-borne bacterial diseases causing fever among malaria smear-negative patients in a rural hospital located in Ethiopia. The study population included patients aged 2 years or older; referred to Gambo Rural General Hospital (West Arsi, Ethiopia), between July and November 2013, for fever or report of fever in the previous 48 h; attending the outpatient department; and testing negative for malaria by Giemsa-stained thin blood smears. We extracted DNA from 394 whole blood samples, using reverse line blot assays of amplicons to look for bacteria from the genera: Anaplasma, Bartonella, Borrelia, Coxiella, Ehrlichia, Francisella, and Rickettsia. Thirteen patients showed presence of DNA for these pathogens: three each by Borrelia spp., the Francisella group (F. tularensis tularensis, F. tularensis holartica, and F. novicia), Rickettsia bellii, and Rickettsia Felis, and one by Bartonella rochalimae. Thus, in this rural area of Africa, febrile symptoms could be due to bacteria transmitted by arthropods. Further studies are needed to evaluate the pathogenic role of R. bellii. [ABSTRACT FROM AUTHOR]

Rickettsioses of the typhus group (TG) and spotted fever group (SFG) are emerging bacterial infections worldwide, especially in the tropics. Only a few studies on these pathogens and their respective clinical diseases have been conducted in Malaysia. Here, we performed a seroprevalence study among 544 healthy, afebrile indigenous people (Orang Asli) from peninsular Malaysia for TG and SFG rickettsioses in nine rural and peri-urban settlements. The study population encompassed children, adolescents, and adults. The overall seroprevalence of rickettsiosis in the Orang Asli was 48.5%, with 27.9% seroprevalence against TG rickettsiae and 20.6% seroprevalence against SFG rickettsiae. In 7.9% of the study participants, antibodies against both rickettsial groups were found. The highest seropositivity rates against TG and SRG rickettsiae were detected in young children and adults. Overall, there were no gender differences. Seroprevalences were similar among inhabitants of different settlements, except for two localities. More studies are needed to shed more light on the ecology and risk factors for TG and SFG rickettsioses in Malaysia. [ABSTRACT FROM AUTHOR]

The present survey aimed to investigate flea and tick fauna parasitizing Slovak red fox populations with special emphasis on canine pathogens they transmit. A total of 407 fleas and 105 ticks were collected from 90 red foxes from two geographically distant regions. Seven flea species (Chaetopsylla globiceps, Pulex irritans, Archaeopsylla erinacei, Chaetopsylla rothschildi, Chaetopsylla trichosa, Ctenocephalides canis, and Ctenopthalmus assimilis) and three species of hard ticks (Ixodes ricinus, Ixodes hexagonus, Haemaphysalis concinna) were recorded on sampled animals. Consequently, the DNA of five different pathogen taxa was confirmed in collected arthropod vectors: Bartonella spp. (in P. irritans, Ch. globiceps, and Ct. assimilis), Rickettsia spp. (in A. erinacei, I. ricinus, I. hexagonus, and H. concinna), Anaplasma phagocytophilum (in I. ricinus), Theileria sp. (in Ch. globiceps and H. concinna), and Hepatozoon canis (in I. ricinus and I. hexagonus). Mycoplasma spp., Dipylidium caninum, and Acanthocheilonema reconditum were not found in fleas or ticks in this study. [ABSTRACT FROM AUTHOR]

Central Queensland (CQ) is a large and isolated, low population density, remote tropical region of Australia with a varied environment. The region has a diverse fauna and several species of ticks that feed upon that fauna. This study examined 518 individual ticks: 177 Rhipicephalus sanguineus (brown dog tick), 123 Haemaphysalis bancrofti (wallaby tick), 102 Rhipicephalus australis (Australian cattle tick), 47 Amblyomma triguttatum (ornate kangaroo tick), 57 Ixodes holocyclus (paralysis tick), 9 Bothriocroton tachyglossi (CQ short-beaked echidna tick), and 3 Ornithodoros capensis (seabird soft tick). Tick midguts were pooled by common host or environment and screened for four genera of tick-borne zoonoses by PCR and sequencing. The study examined a total of 157 midgut pools of which 3 contained DNA of Coxiella burnetii, 13 Rickettsia gravesii, 1 Rickettsia felis, and 4 other Rickettsia spp. No Borrelia spp. or Babesia spp. DNA were recovered. [ABSTRACT FROM AUTHOR]

Dogs are a common feeding hosts for Ixodes ricinus and may act as reservoir hosts for zoonotic tick-borne pathogens (TBPs) and as carriers of infected ticks into human settings. The aim of this work was to evaluate the presence of several selected TBPs of significant public health concern by molecular methods in I. ricinus recovered from dogs living in urban and suburban settings in central Italy. A total of 212 I. ricinus specimens were collected from the coat of domestic dogs. DNA was extracted from each specimen individually and tested for Rickettsia spp., Borrelia burgdorferi sensu lato, Babesia spp., and Anaplasma phagocytophilum, using real-time and conventional PCR protocols, followed by sequencing. Sixty-one ticks (28.8%) tested positive for TBPs; 57 samples were infected by one pathogen, while four showed coinfections. Rickettsia spp. was detected in 39 specimens (18.4%), of which 32 were identified as Rickettsia monacensis and seven as Rickettsia helvetica. Twenty-two samples (10.4%) tested positive for A. phagocytophilum; Borrelia lusitaniae and Borrelia afzelii were detected in two specimens and one specimen, respectively. One tick (0.5%) was found to be positive for Babesia venatorum (EU1). Our findings reveal the significant exposure of dogs to TBPs of public health concern and provide data on the role of dogs in the circulation of I. ricinus-borne pathogens in central Italy. [ABSTRACT FROM AUTHOR]

The genus Rickettsia (Rickettsiales: Rickettsiaceae) includes Gram-negative, small, obligate intracellular, nonmotile, pleomorphic coccobacilli bacteria transmitted by arthropods. Some of them cause human and probably also animal disease (life threatening in some patients). In these guidelines, we give clinical practice advices (microscopy, serology, molecular tools, and culture) for the microbiological study of these microorganisms in clinical samples. Since in our environment rickettsioses are mainly transmitted by ticks, practical information for the identification of these arthropods and for the study of Rickettsia infections in ticks has also been added. [ABSTRACT FROM AUTHOR]