Infection and replication of <Emphasis Type="Italic">Bartonella</Emphasis> species within a tick cell line

Bartonella species are fastidious, gram negative bacteria, some of which are transmitted by arthropod vectors, including fleas, sandflies, and lice. There is very little information regarding the interaction and/or transmission capabilities of Bartonella species by ticks. In the present study, we demonstrate successful infection of the Amblyomma americanum cell line, AAE12, by seven Bartonella isolates and three Candidatus Bartonella species by electron or light microscopy. With the exception of Bartonella bovis, infection with all other examined Bartonella species induced cytopathic effects characterized by heavy cellular vacuolization and eventually cell lysis. Furthermore, using quantitative real time PCR (qPCR), we demonstrated significant amplification of two B. henselae genotype I isolates in the A. americanum cell line over a 5 days period. Ultimately, tick-cell derived Bartonella antigens may prove useful for the development of more sensitive diagnostic reagents and may assist in the development of an effective vaccine to prevent the further spread of disease caused by these organisms.     

Mixed infections, cryptic diversity, and vector-borne pathogens: evidence from Polygenis fleas and Bartonella species

Coinfections within hosts present opportunities for horizontal gene transfer between strains and competitive interactions between genotypes and thus can be a critical element of the lifestyles of pathogens. Bartonella spp. are Alphaproteobacteria that parasitize mammalian erythrocytes and endothelial cells. Their vectors are thought to be various biting arthropods, such as fleas, ticks, mites, and lice, and they are commonly cited as agents of various emerging diseases. Coinfections by different Bartonella strains and species can be common in mammals, but little is known about specificity and coinfections in arthropod vectors. We surveyed the rate of mixed infections of Bartonella in flea vectors (Polygenis gwyni) parasitizing cotton rats (Sigmodon hispidus) in which previous surveys indicated high rates of infection. We found that nearly all fleas (20 of 21) harbored one or more strains of Bartonella, with rates of coinfection approaching 90%. A strain previously identified as common in cotton rats was also common in their fleas. However, another common strain in cotton rats was absent from P. gwyni, while a rare cotton rat strain was quite common in P. gwyni. Surprisingly, some samples were also coinfected with a strain phylogenetically related to Bartonella clarridgeiae, which is typically associated with felids and ruminants. Finally, a locus (pap31) that is characteristically borne on phage in Bartonella was successfully sequenced from most samples. However, sequence diversity in pap31 was novel in the P. gwyni samples, relative to other Bartonella previously typed with pap31, emphasizing the likelihood of large reservoirs of cryptic diversity in natural populations of the pathogen.     

Human bartonellosis: seroepidemiological and clinical features with an emphasis on data from Brazil - a review

Bartonellae are fastidious Gram-negative bacteria that are widespread in nature with several animal reservoirs (mainly cats, dogs, and rodents) and insect vectors (mainly fleas, sandflies, and human lice). Thirteen species or subspecies of Bartonella have been recognized as agents causing human disease, including B. bacilliformis, B. quintana, B. vinsonii berkhoffii, B. henselae, B. elizabethae, B. grahamii, B. washoensis, B. koehlerae, B. rocha-limaea, and B. tamiae. The clinical spectrum of infection includes lymphadenopathy, fever of unknown origin, endocarditis, neurological and ophthalmological syndromes, Carrion's disease, and others. This review provides updated information on clinical manifestations and seroepidemiological studies with an emphasis on data available from Brazil.     

Bartonella and Rickettsia in fleas and lice from mammals in South Carolina, U.S.A.

Species in the genera Bartonella and Rickettsia are vector-borne pathogens of humans and domestic animals. The natural reservoirs and enzootic transmission cycles of these bacteria are poorly known in South Carolina. Thirteen species of lice and fleas were collected from urban animals and screened for the presence of Bartonella and Rickettsia by PCR amplification using genus-specific primers. Bartonella henselae was present in cat fleas (Ctenocephalides felis) from Virginia opossums (Didelphis virginiana) and a novel genotype of Bartonella was detected in Orchopeas howardi from an eastern gray squirrel (Sciurus carolinensis). We detected R. typhi and three novel genotypes Rickettsia in other species of fleas and lice. Rickettsia typhi, the causative agent of murine typhus, was detected in two pools of lice (Enderleinellus marmotae) from the woodchuck (Marmota monax). Cat fleas harbored one of two novel genotypes of Rickettsia. A third novel Rickettsia was detected in Orchopeas howardi from an eastern gray squirrel.     

Ectoparasites of gray squirrels in two different habitats and screening of selected ectoparasites for bartonellae

Gray squirrels, Sciurus carolinensis, were livetrapped in 2 different habitat types, woodland (67 squirrels) and parkland (53 squirrels), in southeastern Georgia. Ectoparasites were recovered from anesthetized squirrels and compared between hosts from the 2 habitats. Because of the absence of low vegetation in parkland habitats, it was hypothesized that the ectoparasite fauna, especially ticks and chiggers, would be more diverse on woodland squirrels. The results were generally in agreement with this hypothesis. Seventeen species of ectoparasites were recovered from woodland squirrels, compared with 6 species from parkland squirrels. Five species of ticks and 3 species of chiggers parasitized the woodland squirrels compared with no ticks or chiggers on the parkland squirrels. Significantly higher infestation prevalences were recorded on woodland compared with parkland squirrels for the flea Orchopeas howardi, the tick Amblyomma americanum, and the mesostigmatid mite Androlaelaps fahrenholzi. The mean intensity for O. howardi also was significantly higher on woodland than on parkland squirrels. Because a new strain of Bartonella sp. was isolated recently from S. carolinensis in Georgia, selected ectoparasites from this study were screened for bartonellae by polymerase chain reaction (PCR). Some of the fleas and lice, but none of the mites tested, were PCR positive, suggesting that fleas, or lice, or both, might be vectors of bartonellae between squirrels. Six distinct strains of Bartonella sp. were detected, 2 in fleas and 4 in lice.     

Bartonellosis,an increasingly recognized zoonosis

Cat scratch disease is the most common zoonotic infection caused by Bartonella bacteria. Among the many mammals infected with Bartonella spp., cats represent a large reservoir for human infection, as they are the main reservoir for Bartonella henselae, Bartonella clarridgeiae and Bartonella koehlerae. Bartonella spp. are vector‐borne bacteria, and transmission of B. henselae by cat fleas occurs mainly through infected flea faeces, although new potential vectors (ticks and biting flies) have been identified. Dogs are also infected with various Bartonella species and share with humans many of the clinical signs induced by these infections. Although the role of dogs as source of human infection is not yet clearly established, they represent epidemiological sentinels for human exposure. Present knowledge on the aetiology, clinical features and epidemiological characteristics of bartonellosis is presented.     

Phylogenetic analysis of Bartonella detected in rodent fleas in Yunnan, China

Previous studies have demonstrated a diversity of Bartonella spp. in rodent populations in Yunnan Province, China. Although Bartonella spp. have been isolated from cat fleas and cattle ticks collected from their animal hosts, little is known about Bartonella carried by rodent fleas. In this study, Bartonella DNA was detected by polymerase chain reaction (PCR) in two of five species of rodent fleas. These included Xenopsylla cheopis and Ctenophthalmus lushuiensis, which were collected from Rattus tanezumi flavipectus and from the nests of voles, respectively, during 1997 from two sites in western Yunnan Province, China. Sequence analysis of the Bartonella citrate synthase gene (gltA) amplicons obtained from six of 65 grouped flea samples showed that Bartonella genetic variants were clustered in four groups. One from Xenopsylla cheopis was identical to Bartonella tribocorum, whereas the other three genotypes from Ctenophthalmus lushuiensis were related to the vole-associated Bartonella isolates and cat-associated Bartonella clarridgeiae. This is the first detection of this Bartonella variant from fleas in China. Therefore, further investigations are needed to clarify the distribution of Bartonella in rodents and their ectoparasites in China to define the role of these arthropods in the transmission routes of Bartonella.     

Saliva activated transmission (SAT) of Thogoto virus: relationship with vector potential of different haematophagous arthropods

Tick saliva (or salivary gland extract) potentiates the transmission of Thogoto (THO) virus to uninfected ticks feeding on a non-viraemic guinea-pig. This phenomenon has been named saliva activated transmission (SAT). To investigate the potential of different haematophagous arthropods to mediate SAT, guinea-pigs were infested with uninfected R.appendiculatus Neumann nymphs and inoculated with THO virus and salivary gland extract (SGE) derived from a range of ixodid (metastriate and prostriate) or argasid ticks, or mosquitoes; control guinea-pigs were inoculated with virus alone. Enhancement of THO virus transmission was observed only when SGE was derived from metastriate ticks. Comparison with the vector potential of these various arthropod species revealed that enhancement of THO virus transmission was specific for ticks which were competent vectors of the virus. The data indicate a correlation between vector competence and the ability of haematophagous arthropods to mediate SAT of THO virus.     

Ectoparasites of the Irish stoat

A sample of 196 Irish stoats Mustela ermineae hibernica Thomas & Barrett-Hamilton (Carnivora: Mustelidae) were examined and yielded 2580 arthropod ectoparasites, including 1819 larval mites Neotrombicula autumnalis (Shaw) from a single host. Other ectoparasites recovered included lice (99% Trichodectes ermineae (Hopkins], ticks (mostly Ixodes hexagonus Leach) and four species of fleas. The ticks and fleas were considered to have come mainly from nests of other hosts. The flea species did not reflect the status of their usual hosts in the diet of stoats, but did reflect the stoats' use of their usual hosts' habitat.     

Investigation of Bartonella acquisition and transmission in Xenopsylla ramesis fleas (Siphonaptera: Pulicidae)

Bartonella are emerging and re-emerging pathogens affecting humans and a wide variety of animals including rodents. Horizontal transmission of Bartonella species by different hematophagous vectors is well acknowledged but vertical transmission (from mother to offspring) is questionable and was never explored in fleas. The aim of this study was to investigate whether the rodent flea, Xenopsylla ramesis, can acquire native Bartonella from wild rodents and transmit it transovarially. For this aim, Bartonella-free laboratory-reared X. ramesis fleas were placed on six naturally Bartonella-infected rodents and six species-matched Bartonella-negative rodents (three Meriones crassus jirds, two Gerbillus nanus gerbils and one Gerbillus dasyurus gerbil) for 7 days, 12-14h per day. The fleas that were placed on the Bartonella-positive rodents acquired four different Bartonella genotypes. Eggs and larvae laid and developed, respectively, by fleas from both rodent groups were collected daily for 7 days and molecularly screened for Bartonella. All eggs and larvae from both groups were found to be negative for Bartonella DNA. Interestingly, two of five gut voids regurgitated by Bartonella-positive fleas contained Bartonella DNA. The naturally infected rodents remained persistently infected with Bartonella for at least 89 days suggesting their capability to serve as competent reservoirs for Bartonella species. The findings in this study indicate that X. ramesis fleas can acquire several Bartonella strains from wild rodents but cannot transmit Bartonella transovarially.     

Mixed Infections, Cryptic Diversity, and Vector-Borne Pathogens: Evidence from Polygenis Fleas and Bartonella Species

Coinfections within hosts present opportunities for horizontal gene transfer between strains and competitive interactions between genotypes and thus can be a critical element of the lifestyles of pathogens. Bartonella spp. are Alphaproteobacteria that parasitize mammalian erythrocytes and endothelial cells. Their vectors are thought to be various biting arthropods, such as fleas, ticks, mites, and lice, and they are commonly cited as agents of various emerging diseases. Coinfections by different Bartonella strains and species can be common in mammals, but little is known about specificity and coinfections in arthropod vectors. We surveyed the rate of mixed infections of Bartonella in flea vectors (Polygenis gwyni) parasitizing cotton rats (Sigmodon hispidus) in which previous surveys indicated high rates of infection. We found that nearly all fleas (20 of 21) harbored one or more strains of Bartonella, with rates of coinfection approaching 90%. A strain previously identified as common in cotton rats was also common in their fleas. However, another common strain in cotton rats was absent from P. gwyni, while a rare cotton rat strain was quite common in P. gwyni. Surprisingly, some samples were also coinfected with a strain phylogenetically related to Bartonella clarridgeiae, which is typically associated with felids and ruminants. Finally, a locus (pap31) that is characteristically borne on phage in Bartonella was successfully sequenced from most samples. However, sequence diversity in pap31 was novel in the P. gwyni samples, relative to other Bartonella previously typed with pap31, emphasizing the likelihood of large reservoirs of cryptic diversity in natural populations of the pathogen.     

The arthropod,but not the vertebrate host or its environment,dictates bacterial community composition of fleas and ticks

Bacterial community composition in blood-sucking arthropods can shift dramatically across time and space. We used 16S rRNA gene amplification and pyrosequencing to investigate the relative impact of vertebrate host-related, arthropod-related and environmental factors on bacterial community composition in fleas and ticks collected from rodents in southern Indiana (USA). Bacterial community composition was largely affected by arthropod identity, but not by the rodent host or environmental conditions. Specifically, the arthropod group (fleas vs ticks) determined the community composition of bacteria, where bacterial communities of ticks were less diverse and more dependent on arthropod traits—especially tick species and life stage—than bacterial communities of fleas. Our data suggest that both arthropod life histories and the presence of arthropod-specific endosymbionts may mask the effects of the vertebrate host and its environment.     

Detection of a malaria parasite (Plasmodium mexicanum) in ectoparasites (mites and ticks), and possible significance for transmission

Two species of sandflies (Lutzomyia) are competent vectors of Plasmodium mexicanum, a malaria parasite of lizards. The very patchy distribution of sites with high P. mexicanum prevalence in the lizards, and often low or even nil sandfly density at such sites, provoked an evaluation of 2 common lizard ectoparasites, the tick Ixodes pacificus and the mite Geckobiella occidentalis, as potential passive vectors. Plasmodium sp.-specific polymerase chain primers were used to amplify a long segment of the mitochondrial cytochrome b gene that is unlikely to survive intact if the parasite cells are killed within a blood-feeding arthropod. The segment was strongly amplified from sandflies (the positive control for the method) from 1 to 96 hr postfeeding on an infected lizard. For ticks, the gene fragment was poorly amplified at 0 hr postfeed, and not amplified after 2 hr. In contrast, strong amplification of the parasite DNA was observed from mites from 0 to 20 hr postfeed, and weak amplification even at 96 hr.     

Vector competence of the tick Ixodes ricinus for transmission of Bartonella birtlesii

Bartonella spp. are facultative intracellular vector-borne bacteria associated with several emerging diseases in humans and animals all over the world. The potential for involvement of ticks in transmission of Bartonella spp. has been heartily debated for many years. However, most of the data supporting bartonellae transmission by ticks come from molecular and serological epidemiological surveys in humans and animals providing only indirect evidences without a direct proof of tick vector competence for transmission of bartonellae. We used a murine model to assess the vector competence of Ixodes ricinus for Bartonella birtlesii. Larval and nymphal I. ricinus were fed on a B. birtlesii-infected mouse. The nymphs successfully transmitted B. birtlesii to naïve mice as bacteria were recovered from both the mouse blood and liver at seven and 16 days after tick bites. The female adults successfully emitted the bacteria into uninfected blood after three or more days of tick attachment, when fed via membrane feeding system. Histochemical staining showed the presence of bacteria in salivary glands and muscle tissues of partially engorged adult ticks, which had molted from the infected nymphs. These results confirm the vector competence of I. ricinus for B. birtlesii and represent the first in vivo demonstration of a Bartonella sp. transmission by ticks. Consequently, bartonelloses should be now included in the differential diagnosis for patients exposed to tick bites.     

Natural breeding places of phlebotomine sandflies

Methods of finding larvae and pupae of phlebotomine sandflies (Diptera: Psychodidae) are described and the known types of breeding sites used by sandflies are listed. Three ways of detecting sandfly breeding places are the use of emergence traps placed over potential sources to catch newly emerged adult sandflies; flotation of larvae and pupae from soil, etc., and desiccation of media to drive out the larvae. Even so, remarkably little information is available on the ecology of the developmental stages of sandflies, despite their importance as vectors of Leishmania, Bartonella and phleboviruses affecting humans and other vertebrates in warmers parts of the world. Regarding the proven or suspected vectors of leishmaniases, information on breeding sites is available for only 15 out of 29 species of sandflies involved in the Old World and 12 out of 44 species of sandflies involved in the Americas, representing approximately 3% of the known species of Phlebotominae. Ecotopes occupied by immature phlebotomines are usually organically rich moist soils, such as the rain forest floor (Lutzomyia intermedia, Lu. umbratilis, Lu. whitmani in the Amazon; Lu. gomezi, Lu. panamensis, Lu. trapidoi in Panama), or contaminated soil of animal shelters (Lu. longipalpis s.l. in South America, Phlebotomus argentipes in India; P. chinensis in China; P. ariasi, P. perfiliewi, P. perniciosus in Europe). Developmental stages of some species (P. langeroni and P. martini in Africa; P. papatasi in Eurasia; Lu. longipalpis s.l. in South America), have been found in a wide range of ecotopes, and many species of sandflies employ rodent burrows as breeding sites, although the importance of this niche is unclear. Larvae of some phlebotomines have been found in what appear to be specialized niches such as Lu. ovallesi on buttress roots of trees in Panama; P. celiae in termite hills in Kenya; P. longipes and P. pedifer in caves and among rocks in East Africa. Old World species found as immatures in the earthen floor of human habitations include P. argentipes, P. chinensis, P. martini and P. papatasi. Much more information on sandfly breeding sites is required to facilitate their control by source reduction.     

The physiology of itch

The perception of itch is associated with many parasites and their vectors, especially following penetration of the skin by the parasites themselves, as in cercarial dermatitis of schistosome infections, or penetration of arthropod mouthparts during blood feeding. Many ectoparasites such as scabies, lice and fleas, provoke sensations of itch - even when the insects are no longer (or have never been) present, giving rise to the phenomenon of delusory parasitosis. Itch, and the host 'grooming' responses with which it is associated, is increasingly recognized as an important factor in modulating vector feeding behaviour, which can have profound effects on the transmission dynamics of vector borne parasites. As a background to future reviews of this developing subject, we asked John Alexander, author of the classic Arthropods and Human Skin (Springer-Verlag, 1984), to explain what is itch, and to discuss what is known about its underlying Physiology.     

Worldwide detection and identification of new and old rickettsiae and rickettsial diseases

To determine the prevalence and distribution of rickettsial pathogens around the world, scientists have relied more and more upon molecular techniques in addition to serological and culture methods. The ease of use and sensitivity/specificity of molecular techniques such as quantitative real-time PCR assays and multilocus sequence typing have lead to an increase in reports of the detection and identification of new and old rickettsiae in previously known and in new endemic regions. These assays have been successfully used with clinical samples such as serum, blood, and tissue biopsies and with environmental samples such as arthropod vectors including ticks, fleas, lice, and mites, and blood and tissue specimens from small mammal collections and from wild and domestic large animals. These methods have lead to the detection of new and old rickettsial pathogens often in new locations leading investigators to suggest new regions of risk of these rickettsioses.     

Association of Bartonella with the fleas (Siphonaptera) of rodents and bats using molecular techniques.

Bartonella spp. are putatively vector-borne bacterial agents of humans and animals. Fleas have been incriminated as vectors of Bartonella spp. and are suspected of transmitting Bartonella of rodents and bats, but some of these Bartonella spp. have not yet been directly detected in wild caught fleas. We report the molecular detection of Bartonella tribocorum, Bartonella vinsonii subsp. vinsonii, and two novel genotypes of Bartonella from the fleas Xenopsylla cheopis, Ctenophthalmus pseudagyrtes, Sternopsylla texanus, or Orchopeas howardi.     

Acaricidal and repellent properties of permethrin, its role in reducing transmission of vector-borne pathogens

Vector-borne pathogens causing canine vector-borne diseases (CVBD) are recognized as being of increasing importance in small animal clinics. The pathogens and their vectors (eg, fleas, ticks, mosquitoes and sand flies) have a global distribution. Their prevalence may vary depending on region, climate, hygiene and many other factors; however the risk of infection for companion animals in our highly developed and supposedly regulated environments will never be zero. Even in highly developed markets, with high socioeconomic standards, ectoparasites are still a threat to both pets and humans. One has to understand the complexity of arthropod biology and esp. the complex mechanisms of host seeking, attachment and skin penetration and finally feeding, to differentiate the various acaricides and their therapeutic as well as prophylactic properties. Prevention of arthropod bites is mainly by prevention of attachment and thus any engorgement if possible. To achieve this, acaricides with repellent properties, such as the synthetic pyrethroid permethrin are ideal compounds to reach this goal.     

Environmental information systems for the control of arthropod vectors of disease

Over the last decade, remote sensing technologies and geographical information systems have moved from the research arena into the hands of vector control specialists. This review explains remote sensing approaches and spatial information technologies used for investigations of arthropod pests and vectors of diseases affecting humans and livestock. Relevant applications are summarized with examples of studies on African horse sickness vector Culicoides midges (Diptera: Ceratopogonidae), malaria vector Anopheles and arbovirus vector culicine mosquitoes (Diptera: Culicidae), leishmaniasis vector Phlebotomus sandflies (Diptera: Psychodidae), trypanosomiasis vector tsetse (Diptera: Glossinidae), loaiasis vector Chrysops (Diptera: Tabanidae), Lyme disease vector Ixodes and other ticks (Acari: Ixodidae). Methods and their uses are tabulated and discussed with recommendations for efficiency, caution and progress in this burgeoning field.