Bartonella species and their ectoparasites: selective host adaptation or strain selection between the vector and the mammalian host?

A wide range of blood-sucking arthropods have either been confirmed or are suspected as important vectors in Bartonella transmission to mammals, including humans. Overall, it appears that the diversity of Bartonella species DNA identified in ectoparasites is much broader than the species detected in their mammalian hosts, suggesting a mechanism of adaptation of Bartonella species to their host-vector ecosystem. However, these mechanisms leading to the fitness between the vectors and their hosts still need to be investigated.     

Bartonellae in animals and vectors in New Caledonia

Bartonellae are gram-negative facultative intracellular alpha-proteobacteria from the family Bartonellaceae. The natural history of bartonellae consists of a reservoir/host, which is a vertebrate with chronic intravascular infection with sustained bacteremia, and a vector (usually an arthropod) that transfers the bacteria from the reservoir to a susceptible yet uninfected host. In order to reveal the sources and reservoirs of Bartonella infection in animals and vectors in New Caledonia, we collected the blood samples of 64 dogs, 8 cats, 30 bovines, 25 horses and 29 wild deer Cervus timorensis russa and 308 associated blood-sucking parasites (14 keds Hippobosca equina, 258 ticks (22 Rhipicephalus microplus, 235 Rhipicephalus sanguineus, and 1 Haemaphysalis longicornis), 12 fleas Ctenocephalides felis and 24 dog lice Trichodectes canis). We isolated ten strains of Bartonella: four Bartonella henselae from cats and six Bartonella chomelii from cattle. The strains were characterized by sequencing of five genes (16S, ITS, rpoB, gltA and ftsZ). The six strains isolated from cattle were close to the reference strain of B. chomelii and were, probably, imported from France with cattle of Limousin race. PCR showed that 35% of keds collected from deer and 31% of deer were infected by B. aff. schoenbuchensis; all other samples were negative. Our data confirmed that in New Caledonia, as in other regions of the world, cats are the major reservoirs of B. henselae. We also confirmed that Hippoboscidae flies may serve as the vectors of ruminant-associated bartonellae.     

Bartonellae are Prevalent and Diverse in Costa Rican Bats and Bat Flies

Species in the bacterial genus, Bartonella, can cause disease in both humans and animals. Previous reports of Bartonella in bats and ectoparasitic bat flies suggest that bats could serve as mammalian hosts and bat flies as arthropod vectors. We compared the prevalence and genetic similarity of bartonellae in individual Costa Rican bats and their bat flies using molecular and sequencing methods targeting the citrate synthase gene (gltA). Bartonellae were more prevalent in bat flies than in bats, and genetic variants were sometimes, but not always, shared between bats and their bat flies. The detected bartonellae genetic variants were diverse, and some were similar to species known to cause disease in humans and other mammals. The high prevalence and sharing of bartonellae in bat flies and bats support a role for bat flies as a potential vector for Bartonella, while the genetic diversity and similarity to known species suggest that bartonellae could spill over into humans and animals sharing the landscape.     

Bartonella,bats and bugs: A review

Ecological, immunological, and epidemiological factors enable bats to transmit an increasingly recognized spectrum of zoonotic agents, and bartonellae are among those emerging pathogens identified in bats and their arthropod ectoparasites. Current data reveal a multifaceted disease ecology where diverse host species distributed around the world interact with a number of Bartonella spp. and several potential vectors. This review summarizes the methods and findings of studies conducted since 2005 to illustrate that Bartonella bacteremia varies by bat species, location, and other potential variables, such as diet with a very high prevalence in hematophagous bats. Among bat families, Bartonella prevalence ranged from 7.3% among Nycteridae to 54.4% in Miniopteridae. Further research can build on these current data to better determine risk factors associated with Bartonella infection in bat populations and the role of their ectoparasites in transmission.     

Experimental infection of dogs with various Bartonella species or subspecies isolated from their natural reservoir

Dogs can be infected by a wide variety of Bartonella species. However, limited data is available on experimental infection of dogs with Bartonella strains isolated from domestic animals or wildlife. We report the inoculation of six dogs with Bartonella henselae (feline strain 94022, 16S rRNA type II) in three sets of two dogs, each receiving a different inoculum dose), four dogs inoculated with B. vinsonii subsp. berkhoffii type I (ATCC strain, one mongrel dog) or type II (coyote strain, two beagles and one mongrel) and B. rochalimae (coyote strain, two beagles). None of the dogs inoculated with B. henselae became bacteremic, as detected by classical blood culture. However, several dogs developed severe necrotic lesions at the inoculation site and all six dogs seroconverted within one to two weeks. All dogs inoculated with the B. v. berkhoffii and B. rochalimae strains became bacteremic at levels comparable to previous experimental infections with either a dog isolate or a human isolate. Our data support that dogs are likely accidental hosts for B. henselae, just like humans, and are efficient reservoirs for both B. v. berkhoffii and B. rochalimae.     

Prevalence of Bartonella species,hemoplasmas, and Rickettsia felis DNA in blood and fleas of cats in Bangkok,Thailand

Flea infestations are common in Thailand, but little is known about the flea-borne infections. Fifty flea pools and 153 blood samples were collected from client-owned cats between June and August 2009 from veterinary hospitals in Bangkok, Thailand. Total DNA was extracted from all samples, and then assessed by conventional PCR assays. The prevalence rates of Bartonella spp. in blood and flea samples were 17% and 32%, respectively, with DNA of Bartonella henselae and Bartonella clarridgeiae being amplified most commonly. Bartonella koehlerae DNA was amplified for the first time in Thailand. Hemoplasma DNA was amplified from 23% and 34% of blood samples and flea pools, respectively, with ‘Candidatus Mycoplasma haemominutum’ and Mycoplasma haemofelis being detected most frequently. All samples were negative for Rickettsia felis. Prevalence rate of B. henselae DNA was increased 6.9 times in cats with flea infestation. Cats administered flea control products were 4.2 times less likely to be Bartonella-infected.     

Molecular and serological diagnosis of Bartonella infection in 61 dogs from the United States

Background: Molecular diagnosis of canine bartonellosis can be extremely challenging and often requires the use of an enrichment culture approach followed by PCR amplification of bacterial DNA. Hypotheses: (1) The use of enrichment culture with PCR will increase molecular detection of bacteremia and will expand the diversity of Bartonella species detected. (2) Serological testing for Bartonella henselae and Bartonella vinsonii subsp. berkhoffii does not correlate with documentation of bacteremia. Animals: Between 2003 and 2009, 924 samples from 663 dogs were submitted to the North Carolina State University, College of Veterinary Medicine, Vector Borne Diseases Diagnostic Laboratory for diagnostic testing with the Bartonellaα‐Proteobacteria growth medium (BAPGM) platform. Test results and medical records of those dogs were retrospectively reviewed. Methods: PCR amplification of Bartonella sp. DNA after extraction from patient samples was compared with PCR after BAPGM enrichment culture. Indirect immunofluorescent antibody assays, used to detect B. henselae and B. vinsonii subsp. berkhoffii antibodies, were compared with PCR. Results: Sixty‐one of 663 dogs were culture positive or had Bartonella DNA detected by PCR, including B. henselae (30/61), B. vinsonii subsp. berkhoffii (17/61), Bartonella koehlerae (7/61), Bartonella volans‐like (2/61), and Bartonella bovis (2/61). Coinfection with more than 1 Bartonella sp. was documented in 9/61 dogs. BAPGM culture was required for PCR detection in 32/61 cases. Only 7/19 and 4/10 infected dogs tested by IFA were B. henselae and B. vinsonii subsp. berkhoffii seroreactive, respectively. Conclusions and Clinical Importance: Dogs were most often infected with B. henselae or B. vinsonii subsp. berkhoffii based on PCR and enrichment culture, coinfection was documented, and various Bartonella species were identified. Most infected dogs did not have detectable Bartonella antibodies.     

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats and dogs in Korea

Blood, saliva, and nail samples were collected from 54 dogs and 151 cats and analyzed for the presence of Bartonella henselae with a novel nested polymerase chain reaction (PCR) method. Bartonella (B.) henselae was detected in feral cat blood (41.8%), saliva (44.1%), and nail (42.7%) samples. B. henselae was also detected in pet cat blood (33.3%), saliva (43.5%), and nail (29.5%) samples and in pet dog blood (16.6%), saliva (18.5%), and nail (29.6%) samples. Nine samples were infected with B. clarridgeiae and 2 were co-infected with B. henselae and B. clarridgeiae of blood samples of dogs. This report is the first to investigate the prevalence of B. henselae and B. clarridgeiae in dogs and cats in Korea, and suggests that dogs and cats may serve as potential Bartonella reservoirs.     

Prevalence of Anaplasma,Bartonella and Borrelia Species in Haemaphysalis longicornis collected from goats in North Korea

North Korea is located on the northern part of the Korean Peninsula in East Asia. While tick-borne pathogens of medical and veterinary importance have been reported from China and South Korea, they have not been reported from North Korea. To screen for zoonotic tick-borne pathogens in North Korea, ticks were collected from domestic goats. A total of 292 (27 nymph, 26 male, 239 female) Haemaphysalis (H.) longicornis were collected and assayed individually for selected tick-borne pathogens. A total of 77 (26.4%) were positive for Anaplasma bovis, followed by Bartonella (B.) grahamii (15, 5.1%), Anaplasma phagocytophilum (12, 4.1%), Bartonella henselae (10, 3.4%), and Borrelia spp. (3, 1.0%) based on 16S ribosomal RNA and ITS species-specific nested polymerase chain reaction. Using the groEL-based nested PCR, a total of 6 and 1 H. longicornis were positive for B. grahamii and B. henselae, respectively. All products were sequenced and demonstrated 100% identity and homology with previously reported sequences from other countries in GenBank. This is the first report of the detection of tick-borne pathogens in the North Korea and suggests that farm animals may act as reservoirs for zoonotic tick-borne pathogens.     

Detection of Bartonella spp. in Cimex lectularius by MALDI-TOF MS

Bed bugs are small hematophagous insects. They are found in temperate and tropical climates around the world. Their vectorial capacity for several pathogens, including Bartonella spp., has been suspected. An experimental study of artificial infection of Cimex lectularius with Bartonella quintana and Bartonella henselae bacteria was developed to evaluate the ability of MALDI-TOF MS to simultaneously identify bed bugs and their infectious status. This experimental study confirmed the ability of MALDI-TOF MS to identify bed bugs. In addition, it was able to differentiate between control bed bugs, bed bugs infected with Bartonella quintana and bed bugs infected with Bartonella henselae, with an identification percentage above 90%.     

The prevalence of Bartonella, hemoplasma, and Rickettsia felis infections in domestic cats and in cat fleas in Ontario

The prevalence of persistent bacteremic Bartonella spp. and hemoplasma infections was determined in healthy pet cats in Ontario. Blood samples from healthy cats sent to a diagnostic laboratory for routine health assessment over the course of 1 y were tested for Bartonella spp. using both polymerase chain reaction (PCR) and blood culture, and for the presence of hemoplasma by PCR. The overall prevalence of Bartonella spp. by PCR and by culture combined was 4.3% (28/646) [3.7% (24/646) Bartonella henselae, 0.6% (4/646) Bartonella clarridgeiae]. The novel B. henselae PCR developed for this study demonstrated nearly twice the sensitivity of bacterial isolation. The overall prevalence of hemoplasma was 4% (30/742) [3.3% (25/742) Candidatus Mycoplasma haemominutum, 0.7% (5/742) Mycoplasma haemofelis]. There was no significant difference between the prevalence of infection by season or by age (≤ 2 y, > 2 y). Candidatus Mycoplasma turicensis was identified, for the first time in Canada, in 1 cat. The prevalence of Bartonella (58%) and hemoplasma (47% M. haemofelis, 13% M. haemominutum) in blood from a small sampling (n = 45) of stray cats was considerably higher than that found in healthy pet cats.     

Analysis of Seroreactivity against Cell Culture–Derived Bartonella spp. Antigens in Dogs

Background

Little is known about the specificity of Bartonella spp. immunofluorescent antibody (IFA) assays in dogs. Bacteremia in sick dogs most often has been associated with Bartonella henselae (Bh), Bartonella vinsonii subspecies berkhoffii (Bvb), and Bartonella koehlerae (Bk). Clarification of the diagnostic utility of IFA serology when testing against these organisms is needed.

Objective

To evaluate the specificity of Bartonella IFA assays utilizing 6 cell culture–grown antigen preparations.

Animals

Archived sera from SPF dogs (n = 29) and from dogs experimentally infected with Bvb (n = 10) and Bh (n = 3).

Methods

Antibodies (Abs) to Bvb genotypes I, II, and III, Bh serotype I, strains H‐1 and SA2, and to Bk were determined by IFA testing.

Results

Serum from naïve SPF dogs shown to be negative for Bartonella bacteremia did not react with any of the 6 Bartonella antigens by IFA testing. Dogs experimentally infected with Bvb genotype I developed Abs against homologous antigens, with no cross‐reactivity to heterologous Bvb genotypes, Bh H‐1, SA2 strains, or to Bk. Dogs experimentally infected with Bh serotype I developed Abs against Bh H‐1, but not to Bh SA2 strain with no cross‐reactive Abs to Bvb genotypes I–III or to Bk.

Conclusions and Clinical Importance

Bartonella spp. Ab responses during acute experimental infections are species and type specific.     

Bartonella henselae infection in a dog with recalcitrant ineffective erythropoiesis

Ineffective erythropoiesis was diagnosed in an 8‐year‐old male castrated Labrador Retriever. Despite treatment with immunosuppressive therapy for suspected immune‐mediated erythrocyte maturation arrest, resolution of the nonregenerative anemia was not achieved. Following documentation of Bartonella henselae bacteremia by Bartonella alpha proteobacteria growth medium (BAPGM) enrichment blood culture, immunosuppressive therapy was discontinued, and the anemia resolved following prolonged antibiotic therapy. Bartonella immunofluorescent antibody testing was negative, whereas B henselae western blot was consistently positive. The contribution of B henselae bacteremia to ineffective erythropoiesis remains unknown; however, the potential role of B henselae in the pathophysiology of bone marrow dyscrasias warrants additional investigation.     

Flea-borne pathogens in the cat flea Ctenocephalides felis and their association with mtDNA diversity of the flea host

Flea-borne pathogens were screened from 100 individual cat fleas using a PCR approach, of which 38 % were infected with at least one bacterium. Overall, 28 % of the flea samples were positive for Bartonella as inferred from ITS DNA region. Of these, 25 % (7/28) were identified as Bartonella clarridgeiae, 42.9 % (12/28) as Bartonella henselae consisted of two different strains, and 32.1 % (9/28) as Bartonella koehlerae, which was detected for the first time in Malaysia. Sequencing of gltA amplicons detected Rickettsia DNA in 14 % of cat flea samples, all of them identified as Rickettsia asembonensis (100 %). None of the flea samples were positive for Mycoplasma DNA in 16S rRNA gene detection. Four fleas were co-infected with Bartonella and Rickettsia DNAs. Statistical analyses reveal no significant association between bacterial infection and mtDNA diversity of the cat flea. Nevertheless, in all types of pathogen infections, infected populations demonstrated lower nucleotide and haplotype diversities compared to uninfected populations. Moreover, lower haplotype numbers were observed in infected populations.     

Bartonella Seroepidemiology in Dogs from North America, 2008–2014

Background

Improved understanding of Bartonella species seroepidemiology in dogs may aid clinical decision making and enhance current understanding of naturally occurring arthropod vector transmission of this pathogen.

Objectives

To identify demographic groups in which Bartonella exposure may be more likely, describe spatiotemporal variations in Bartonella seroreactivity, and examine co‐exposures to other canine vector‐borne diseases (CVBD).

Animals

A total of 15,451 serology specimens from dogs in North America were submitted to the North Carolina State University, College of Veterinary Medicine Vector Borne Disease Diagnostic Laboratory between January 1, 2008, and December 31, 2014.

Methods

Bartonella henselae, Bartonella koehlerae, and Bartonella vinsonii subspecies berkhoffii indirect fluorescent antibody (IFA) serology results, as well as results from a commercial assay kit screening for Dirofilaria immitis antigen and Ehrlichia species, Anaplasma phagocytophilum, and Borrelia burgdorferi antibodies, and Ehrlichia canis, Babesia canis, Babesia gibsoni, and Rickettsia species IFA results were reviewed retrospectively.

Results

Overall, 3.26% of dogs were Bartonella spp. seroreactive; B. henselae (2.13%) and B. koehlerae (2.39%) were detected more frequently than B. vinsonii subsp. berkhoffii (1.42%, P < 0.0001). Intact males had higher seroreactivity (5.04%) than neutered males (2.87%, P < 0.0001) or intact or spayed females (3.22%, P = 0.0003). Mixed breed dogs had higher seroreactivity (4.45%) than purebred dogs (3.02%, P = 0.0002). There was no trend in seasonal seroreactivity; geographic patterns supported broad distribution of exposure, and co‐exposure with other CVBD was common.

Conclusions and Clinical Importance

Bartonella spp. exposure was documented throughout North America and at any time of year. Male intact dogs, mixed breed dogs, and dogs exposed to other CVBD have higher seroreactivity to multiple Bartonella species.     

Bartonella spp. in cats from Buenos Aires,Argentina

In Argentina, data on the presence of members of the genus Bartonella is scarce. To increase knowledge about these zoonotic pathogens in this country, the presence and variability of Bartonella spp. was investigated in cats and dogs from Buenos Aires. Bartonella spp. was detected in 17.8% of cats, while all dogs tested negative by PCR and Reverse Line Blot. B. henselae was the most frequent species, being detected in 11.9% (14/101), while B. clarridgeiae was found in only 5.9% (6/101) of the cats. Afterwards, B. henselae isolates and positive blood samples were characterized by Multiple Locus Sequence Typing (MLST) and Multiple Locus Variable Number Tandem Repeats Analysis (MLVA). As result, four different MLST sequence types (ST) and eight MLVA profiles were identified. ST 1 was the most frequent variant found in cats, followed by ST 8. Interestingly, some of the MLVA profiles that were detected in this study have been previously associated with human disease, and represents a potential risk of infection. Veterinarians and physicians should consider the presence of these emerging pathogens in their diagnostic routine.     

Co-isolation of Bartonella henselae and Bartonella vinsonii subsp. berkhoffii from blood, joint and subcutaneous seroma fluids from two naturally infected dogs

This report describes the clinical presentation, isolation and treatment of two dogs naturally infected with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii. Chronic and progressive polyarthritis was the primary complaint for dog #1, from which B. henselae and B. vinsonii subsp. berkhoffii were cultured on three independent occasions from blood and joint fluid samples, despite administration of nearly 4 months of non-consecutive antibiotic therapy. A clinically atypical and progressively severe trauma-associated seroma was the primary complaint for dog #2, from which B. henselae and B. vinsonii subsp. berkhoffii were isolated from serum, blood and seroma fluid. Dogs can be co-infected with two Bartonella spp. and infection with these organisms should not be ruled out if specific antibodies are not detected. Specialized culture techniques should be used for isolation and to assess antibiotic efficacy.     

Dogs are more permissive than cats or guinea pigs to experimental infection with a human isolate of Bartonella rochalimae

Bartonella rochalimae was first isolated from the blood of a human who traveled to Peru and was exposed to multiple insect bites. Foxes and dogs are likely natural reservoirs for this bacterium. We report the results of experimental inoculation of two dogs, five cats and six guinea pigs with the only human isolate of this new Bartonella species. Both dogs became bacteremic for 5–7 weeks, with a peak of 10³–10⁴ colony forming units (CFU)/mL blood. Three cats had low bacteremia levels (< 200 CFU/mL) of 6–8 weeks’ duration. One cat that remained seronegative had two bacterial colonies isolated at a single culture time point. A fifth cat never became bacteremic, but seroconverted. None of the guinea pigs became bacteremic, but five seroconverted. These results suggest that dogs could be a reservoir of this strain of B. rochalimae, in contrast to cats and guinea pigs.     

Molecular detection of Bartonella henselae and Bartonella clarridgeiae in clinical samples of pet cats from Southern Italy

Bartonella henselae is considered an emerging pathogen of veterinary and medical interest that can be occasionally transmitted to humans. Cats are considered to be the only reservoir host for B. henselae. In this study, we used a nested-PCR assay to investigate the prevalence of B. henselae and Bartonella clarridgeiae DNA in peripheral blood samples, fine needle lymph node aspirate specimens and oral swabs from 85 cats in order to develop an easy diagnostic strategy for the selection of infection-free cats that are being considered as pets, especially for immunocompromised patients. Overall, molecular analysis showed that 71 cats (83.5%) tested PCR positive for the presence of B. henselae DNA. PCR amplification of DNA B. henselae produced positive products from lymph node aspirate specimens (62/85; 72.9%) similar to those obtained from blood samples (60/85; 70.6%) and higher than those from oral swabs (51/85; 60%) of cats. No PCR product was obtained for B. clarridgeiae. The simultaneous analysis of three different clinical samples in our study increased the diagnostic possibilities for B. henselae infection in the examined cats from 60–72.9% to 83.5%. Lymph node aspirates were found to be the most effective clinical samples for the detection of B. henselae and blood samples were the next best. Oral swab samples were used in this study with good results when considered in combination with blood and/or lymph node aspiration. The use of nested-PCR assay on these three clinical samples may enhance the diagnostic sensitivity for bartonellosis in cats irrespective of the clinical status of animals.     

Babesia and its hosts: adaptation to long-lasting interactions as a way to achieve efficient transmission

Babesia, the causal agent of babesiosis, are tick-borne apicomplexan protozoa. True babesiae (Babesia genus sensu stricto) are biologically characterized by direct development in erythrocytes and by transovarial transmission in the tick. A large number of true Babesia species have been described in various vertebrate and tick hosts. This review presents the genus then discusses specific adaptations of Babesia spp. to their hosts to achieve efficient transmission. The main adaptations lead to long-lasting interactions which result in the induction of two reservoirs: in the vertebrate host during low long-term parasitemia and throughout the life cycle of the tick host as a result of transovarial and transstadial transmission. The molecular bases of these adaptations in vertebrate hosts are partially known but few of the tick-host interaction mechanisms have been elucidated.