Zoonotic vector-borne bacteria in wild rodents and associated ectoparasites from Tunisia.

Wild rodents are considered as potential carriers of several zoonotic vector-borne bacteria but their epidemiology is poorly understood in Tunisia. A total of 305 biological samples (100 spleens, 100 livers, 100 kidneys, and 5 pooled ectoparasites (Xenopsylla cheopis, Laelaps echidninus, Ornithonyssus sp., Hoplopleura sp. and eggs of the rat fleas)) were collected from 100 wild rodents from three Tunisian governorates. Molecular screening was performed to reveal infections with main vector-borne bacteria. Captured rodents belonged to three rodent genera and species including Rattus rattus (n = 51, 51%), Meriones shawi (n = 24, 24%) and Mus musculus (n = 25, 25%). Examined rodents were found to be heavily infested by the rat flea X. cheopis (n = 32, 47%) and the rat mite L. echidninus (n = 22, 32.3%). However, the rat mite Ornithonyssus sp. (n = 13, 19.1%) and the rat lice Hoplopleura sp. (n = 1, 1.5%) were rarely identified. Based on 16S rRNA and msp4 genes, infection with Anaplasmataceae bacteria was detected in six specimens of R. rattus and one M. shawi. Pathogenic A. phagocytophilum (n = 1), A. phagocytophilum-like 1 (Anaplasma sp. Japan) (n = 1), and A. ovis (n = 5) were identified. On the basis of ompB, ompA and gltA genes, infection with Rickettsia spp. was identified in three specimens of R. rattus and one of M. shawi. Five Rickettsia species of the spotted fever group, corresponding to R. monacensis, R. helvetica, R. massiliae, R. africae, and R. aeschlimannii, were detected in mixed infections. Bartonella henselae DNA was also found in two R. rattus, based on rpoB partial sequences. All revealed Anaplasma, Rickettsia and Bartonella bacteria were detected in spleen samples. Ehrlichia, Coxiella and Borrelia spp. were not identified in any of the tested samples. In Tunisia, this is the first report indicating infections with Anaplasma, Rickettsia and Bartonella spp. in wild rodents, particularly present alongside domestic livestock and human. This represents a serious risk of potential bacterial transmission. Thus, controlling rodent population in animal herds, residential areas and sensitizing local people to this risk seem absolutely necessary.

Adaptation of gltA and ssrA assays for diversity profiling by Illumina sequencing to identify Bartonella henselae, B. clarridgeiae and B. koehlerae.

Introduction. Bartonellosis is an emerging zoonotic disease caused by bacteria of the genus Bartonella. Mixed Bartonella infections are a well-documented phenomenon in mammals and their ectoparasites. The accurate identification of Bartonella species in single and mixed infections is valuable, as different Bartonella species have varying impacts on infected hosts.Gap Statement. Current diagnostic methods are inadequate at identifying the Bartonella species present in mixed infections.Aim. The aim of this study was to adopt a Next Generation Sequencing (NGS) approach using Illumina sequencing technology to identify Bartonella species and demonstrate that this approach can resolve mixed Bartonella infections.Methodology. We used Illumina PCR amplicon NGS to target the ssrA and gltA genes of Bartonella in fleas collected from cats, dogs and a hedgehog in Israel. We included artificially mixed Bartonella samples to demonstrate the ability for NGS to resolve mixed infections and we compared NGS to traditional Sanger sequencing.Results. In total, we identified 74 Ctenocephalides felis, two Ctenocephalides canis, two Pulex irritans and three Archaeopsylla e. erinacei fleas. Real-time PCR of a subset of 48 fleas revealed that twelve were positive for Bartonella, all of which were cat fleas. Sanger sequencing of the ssrA and gltA genes confirmed the presence of Bartonella henselae, Bartonella clarridgeiae and Bartonella koehlerae. Illumina NGS of ssrA and gltA amplicons further confirmed the Bartonella species identity in all 12 flea samples and unambiguously resolved the artificially mixed Bartonella samples.Conclusion. The adaptation and multiplexing of existing PCR assays for diversity profiling via NGS is a feasible approach that is superior to traditional Sanger sequencing for Bartonella speciation and resolving mixed Bartonella infections. The adaptation of other PCR primers for Illumina NGS will be useful in future studies where mixed bacterial infections may be present.

Molecular investigation of bacterial and protozoal pathogens in ticks collected from different hosts in Turkey.

BACKGROUND: The emergence of tick-borne disease is increasing because of the effects of the temperature rise driven by global warming. In Turkey, 19 pathogens transmitted by ticks to humans and animals have been reported. Based on this, this study aimed to investigate tick-borne pathogens including Hepatozoon spp., Theileria spp., Babesia spp., Anaplasma spp., Borrelia spp., and Bartonella spp. in tick samples (n = 110) collected from different hosts (dogs, cats, cattle, goats, sheep, and turtles) by molecular methods. METHODS: To meet this objective, ticks were identified morphologically at the genus level by microscopy; after DNA isolation, each tick sample was identified at the species level using the molecular method. Involved pathogens were then investigated by PCR method. RESULTS: Seven different tick species were identified including Rhipicephalus sanguineus, R. turanicus, R. bursa, Hyalomma marginatum, H. anatolicum, H. aegyptium, and Haemaphysalis erinacei. Among the analyzed ticks, Hepatozoon spp., Theileria spp., Babesia spp., and Anaplasma spp. were detected at rates of 6.36%, 16.3%, 1.81%, and 6.36%, respectively while Borrelia spp. and Bartonella spp. were not detected. Hepatozoon spp. was detected in R. sanguineus ticks while Theileria spp., Babesia spp., and Anaplasma spp. were detected in R. turanicus and H. marginatum. According to the results of sequence analyses applied for pathogen positive samples, Hepatozoon canis, Theileria ovis, Babesia caballi, and Anaplasma ovis were identified. CONCLUSION: Theileria ovis and Anaplasma ovis were detected for the first time to our knowledge in H. marginatum and R. turanicus collected from Turkey, respectively. Also, B. caballi was detected for the first time to our knowledge in ticks in Turkey.

Bartonella species as a cause of culture-negative endocarditis in South Africa.

Previous reports have highlighted the high prevalence of blood culture negative endocarditis (BCNE) in South Africa. The Tygerberg Endocarditis Cohort (TEC) study is an ongoing prospective cohort study of patients with confirmed or suspected IE presenting to Tygerberg Academic Hospital, Cape Town, South Africa. Current analysis includes patients that presented between November 2019 and August 2020. Forty four (44) patients have been included in this ongoing study. Fourteen of the 44 patients (31.8%) had BCNE. Further analysis of the patients with BCNE identified Bartonella species as the most common causative organism (n=6; 43%). Other causes included Mycoplasma species (n=2). No cause could be identified in 4 of the 44 patients (9%). Bartonella quintana was identified with PCR of valvular tissue as the causative organism in 4 of the 5 patients that underwent urgent surgery. The patients with Bartonella IE (n=6) had an average age of 39 years with equal gender distribution. The common clinical features were clubbing (n=5; 83%), anemia (n=4; 66.6%), haematuria (n=3; 50%), acute on chronic severe regurgitant lesion (n=3; 50%) and acute severe regurgitant lesion (n=2; 33.3%).The aortic valve was involved in 5 of 6 patients. During a mean follow-up period of 251 days after diagnosis, no major adverse events occurred. Bartonella-associated IE is an important cause of BCNE in the Western Cape of South Africa. Imaging findings (in patients with BCNE) of significant valvular destruction with large vegetations on the aortic valve not affected by congenital or rheumatic valve disease should raise the suspicion of Bartonella-associated IE.

Zoonotic Bartonella species in Eurasian wolves and other free-ranging wild mammals from Italy.

Bartonellae are emerging vector-borne pathogens infecting humans, domestic mammals and wildlife. Ninety-seven red foxes (Vulpes vulpes), 8 European badgers (Meles meles), 6 Eurasian wolves (Canis lupus), 6 European hedgehogs (Erinaceus europaeus), 3 beech martens (Martes foina) and 2 roe deer (Capreolus capreolus) from Italian Nature Conservatory Parks were investigated for Bartonella infection. Several Bartonella species (9.84%; 95% CI: 4.55-15.12), including zoonotic ones, were molecularly detected among wolves (83.3%; 95% CI: 51-100.00), foxes (4.12%; 95% CI: 0.17-8.08), hedgehogs (33.33%; 95% CI: 0.00-71.05) and a roe deer. Bartonella rochalimae was the most common Bartonella species (i.e. in 4 foxes and 2 wolves) detected. Candidatus B. merieuxii and B. vinsonii subsp. berkhoffii were identified for the first time in wolves. Furthermore, Bartonella schoenbuchensis was identified in a roe deer and a new clone with phylogenetic proximity to B. clarridgeiae was detected in European hedgehogs. Zoonotic and other Bartonella species were significantly more frequent in Eurasian wolves (p < .0001), than in other free-ranging wild mammals, representing a potential reservoir for infection in humans and domestic animals.

Bats are key hosts in the radiation of mammal-associated Bartonella bacteria.

Bats are notorious reservoirs of several zoonotic diseases and may be uniquely tolerant of infection among mammals. Broad sampling has revealed the importance of bats in the diversification and spread of viruses and eukaryotes to other animal hosts. Vector-borne bacteria of the genus Bartonella are prevalent and diverse in mammals globally and recent surveys have revealed numerous Bartonella lineages in bats. We assembled a sequence database of Bartonella strains, consisting of nine genetic loci from 209 previously characterized Bartonella lineages and 121 new cultured isolates from bats, and used these data to perform a comprehensive phylogenetic analysis of the Bartonella genus. This analysis included estimation of divergence dates using a molecular clock and ancestral reconstruction of host associations and geography. We estimate that Bartonella began infecting mammals 62 million years ago near the Cretaceous-Paleogene boundary. Additionally, the radiation of particular Bartonella clades correlate strongly to the timing of diversification and biogeography of mammalian hosts. Bats were inferred to be the ancestral hosts of all mammal-associated Bartonella and appear to be responsible for the early geographic expansion of the genus. We conclude that bats have had a deep influence on the evolutionary radiation of Bartonella bacteria and their spread to other mammalian orders. These results support a 'bat seeding' hypothesis that could explain similar evolutionary patterns in other mammalian parasite taxa. Application of such phylogenetic tools as we have used to other taxa may reveal the general importance of bats in the ancient diversification of mammalian parasites.

Lesions associated with Bartonella taylorii-like bacterium infection in a free-ranging, young-of-the-year raccoon from Prince Edward Island, Canada.

A young-of-the year, female raccoon was presented with clinical signs of weakness and tremors. The raccoon was euthanized, and autopsy findings included poor body condition, diffuse lymphadenopathy, and pale, firm kidneys with petechial hemorrhages throughout the renal cortex. Histologic lesions included systemic fibrinoid vascular necrosis and severe renal lesions, including lymphoplasmacytic interstitial nephritis and fibrinosuppurative glomerulonephritis. Inflammatory vascular lesions were also present within the uvea, heart, lymph nodes, and the lamina propria of the gastric wall. Ancillary testing was negative for Borrelia burgdorferi, Leptospira sp., Aleutian disease virus, canine distemper virus, feline coronavirus, porcine circovirus 2, and rabies virus. Transmission electron microscopy revealed large numbers of ~1.3 × 0.35 µm bacterial rods surrounded by a trilaminar cell wall located within the glomeruli and associated with aggregates of fibrin and vascular damage. Analysis of partial citrate synthase gene and 16S-23S ribosomal RNA intergenic spacer region sequences from kidney tissue confirmed that the organism was a Bartonella spp. that was related to numerous Bartonella spp. from shrews in Europe. This group formed a sister clade to the genetically diverse Bartonella taylorii group that has been reported from a wide range of Eurasian rodent and flea species.

Genetic diversity of Bartonella species in small mammals in the Qaidam Basin, western China.

Investigation of the prevalence and diversity of Bartonella infections in small mammals in the Qaidam Basin, western China, could provide a scientific basis for the control and prevention of Bartonella infections in humans. Accordingly, in this study, small mammals were captured using snap traps in Wulan County and Ge'ermu City, Qaidam Basin, China. Spleen and brain tissues were collected and cultured to isolate Bartonella strains. The suspected positive colonies were detected with polymerase chain reaction amplification and sequencing of gltA, ftsZ, RNA polymerase beta subunit (rpoB) and ribC genes. Among 101 small mammals, 39 were positive for Bartonella, with the infection rate of 38.61%. The infection rate in different tissues (spleens and brains) (χ2 = 0.112, P = 0.738) and gender (χ2 = 1.927, P = 0.165) of small mammals did not have statistical difference, but that in different habitats had statistical difference (χ2 = 10.361, P = 0.016). Through genetic evolution analysis, 40 Bartonella strains were identified (two different Bartonella species were detected in one small mammal), including B. grahamii (30), B. jaculi (3), B. krasnovii (3) and Candidatus B. gerbillinarum (4), which showed rodent-specific characteristics. B. grahamii was the dominant epidemic strain (accounted for 75.0%). Furthermore, phylogenetic analysis showed that B. grahamii in the Qaidam Basin, might be close to the strains isolated from Japan and China. Overall, we observed a high prevalence of Bartonella infection in small mammals in the Qaidam Basin. B. grahamii may cause human disease, and the pathogenicity of the others Bartonella species needs further study, the corresponding prevention and control measures should be taken into consideration.

Lipoptena fortisetosa as a vector of Bartonella bacteria in Japanese sika deer (Cervus nippon).

BACKGROUND: Two species of deer ked (Lipoptena cervi and L. mazamae) have been identified as vectors of Bartonella bacteria in cervids in Europe and the USA. In an earlier study we showed that Japanese sika deer (Cervus nippon) harbor three Bartonella species, namely B. capreoli (lineage A) and two novel Bartonella species (lineages B and C); however, there is currently no information on the vector of Bartonella bacteria in sika deer. The aim of this study was to clarify potential vectors of Bartonella in Japanese sika deer. METHODS: Thirty-eight wingless deer keds (L. fortisetosa) and 36 ticks (Haemaphysalis and Ixodes species) were collected from sika deer. The prevalence of Bartonella in the arthropods was evaluated by real-time PCR targeting the 16S-23S internal transcribed spacer (ITS) and by culture of the organisms. The total number of Bartonella bacteria were quantified using real-time PCR. The distribution of Bartonella bacteria in deer ked organs was examined by immunofluorescence analysis. The relationship of Bartonella strains isolated from sika deer and arthropods were examined by a phylogenetic analysis based on concatenated sequences of the gltA, rpoB, ftsZ, and ribC genes, followed by a BLAST search for gltA and rpoB. RESULTS: Bartonella prevalence in deer keds was 87.9% by real-time PCR and 51.5% in culture and that in the ticks was 8.3% by real-time PCR and 2.8% in culture. The mean number of Bartonella bacteria per ked was calculated to be 9.2 × 105 cells. Bartonella aggregates were localized in the midgut of the keds. The phylogenetic analysis and BLAST search showed that both the host deer and the keds harbored two Bartonella species (lineages B and C), while B. capreoli (lineage A) was not detected in the keds. Two novel Bartonella species (lineages D and E) were isolated from one ked. CONCLUSIONS: Lipoptena fortisetosa likely serves as a vector of at least two Bartonella species (lineages B and C), whereas ticks do not seem to play a significant role in the transmission of Bartonella between sika deer based on the lower detection rates of Bartonella in ticks compared to keds. Bartonella species in lineages D and E appear to be L. fortisetosa-specific strains.

Assessment of tick-borne pathogens presence in Dermacentor reticulatus ticks in north-eastern Poland.

PURPOSE: Dermacentor reticulatus is the second most common tick species in Poland after Ixodes ricinus. The aim of the study was to analyze the presence of pathogen DNA in D. reticulatus. MATERIALS AND METHODS: Ticks were collected in The Protected Landscape Area of the Bug and Nurzec Valley (52°40' N and 22°28' E) between 2016 and 2017. End-point PCR for Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia spp., Rickettsia spp., Bartonella spp. and Coxiella burnetii detection was performed. RESULTS: Tick-borne pathogens' DNA was detected in 11.3% of 301 ticks: B. burgdorferi s.l. in 3.6%, Babesia spp. in 6.3%, A. phagocytophilum in 0.7% and B. burgdorferi s.l.-Babesia spp. co-infection in 0.7%. In all 21 Babesia spp. positive samples, sequence analysis confirmed the presence of Babesia canis with an 80.3%-98.3% homology with the B. canis sequences in GenBank. C. burnetii, Bartonella spp., and Rickettsia spp. DNA were not detected. CONCLUSIONS: Dermacentor reticulatus from north-eastern Poland were found to carry three of the most common tick-borne pathogens (B. burgdorferi s.l., Babesia canis, A. phagocytophilum) which lead to single and mixed infections. Babesia canis was the most prevalent pathogen identified in D. reticulatus.

Detection of pathogens in ixodid ticks collected from animals and vegetation in five regions of Ukraine.

The distribution and prevalence of zoonotic pathogens infecting ixodid ticks in Western Europe have been extensively examined. However, data on ticks and tick-borne pathogens in Eastern Europe, particularly Ukraine are scarce. The objective of the current study was, therefore, to investigate the prevalence of Anaplasma phagocytophilum, Anaplasmataceae, Rickettsia spp., Babesia spp., Bartonella spp., and Borrelia burgdorferi sensu lato in engorged and questing ixodid ticks collected from five administrative regions (oblasts) of Ukraine, namely Chernivtsi, Khmelnytskyi, Kyiv, Ternopil, and Vinnytsia. The ticks were collected from both wild and domestic animals and from vegetation. Of 524 ixodid ticks collected, 3, 99, and 422 ticks were identified as Ixodes hexagonus, Ixodes ricinus, and Dermacentor reticulatus, respectively. DNA samples individually extracted from 168 questing and 354 engorged adult ticks were subjected to pathogen-specific PCR analyses. The mean prevalence in I. ricinus and D. reticulatus were, respectively: 10 % (10/97) and 3 % (12/422) for A. phagocytophilum; 69 % (67/97) and 52 % (220/422) for members of the Anaplasmataceae family; 25 % (24/97) and 28 % (117/422) for Rickettsia spp.; 3 % (3/97) and 1 % (6/422) for Babesia spp.; and 9 % (9/97) and 5 % (20/422) for Bartonella spp. Overall, between the five cities, there was no significant difference in the prevalence of any of the pathogens for the respective ticks (p > 0.05). The prevalence of B. burgdorferi s. l. in the questing and engorged I. ricinus varied from 0 to 27 % and 14-44%, respectively, with no statistical significance identified between the five cities (p > 0.05). In addition to reporting the updated data for Kyiv and Ternopil, this study is the first to provide the prevalences of the tick-borne pathogens for Chernivtsi, Khmelnytskyi, and Vinnytsia. This investigation is also the first to detect Neoehrlichia mikurensis in ixodid ticks from Ukraine. These new data will be useful for medical and veterinary practitioners as well as public health officials when diagnosing infections and when implementing measures to combat tick-borne diseases in Ukraine.

Molecular and Serological Survey of the Cat-Scratch Disease Agent (Bartonella henselae) in Free-Ranging Leopardus geoffroyi and Leopardus wiedii (Carnivora: Felidae) From Pampa Biome, Brazil.

The genus Bartonella comprises emerging bacteria that affect humans and other mammals worldwide. Felids represent an important reservoir for several Bartonella species. Domestic cats are the main reservoir of Bartonella henselae, the agent of cat scratch disease (CSD). It can be transmitted directly by scratches and bites from infected cats and via cat fleas. This study aims to investigate the circulation of Bartonella spp. in free-ranging Neotropical wild felids from Southern Brazil using serological and molecular methods. In this study, 53 live-trapped free-ranging wild felids were sampled, 39 Leopardus geoffroyi and 14 Leopardus wiedii, from five municipalities in the Rio Grande, do Sul state, southern Brazil. All captured animals were clinically healthy. Two blood samples of L. geoffroyi were positive, by PCR, for the presence of B. henselae DNA. Conversely, none of L. wiedii blood samples were positive when tested using PCR. Indirect immunofluorescence assay (IFA) showed that 28% of serum samples of wild felids were reactive (seropositive) for B. henselae by immunofluorescence, with titers ranging from 64 to 256. The results presented here provide the first evidence of a Bartonella-enzootic cycle involving L. geoffroyi and L. wiedii, which may account for the spillover of the emerging zoonotic pathogen B. henselae for the indigenous fauna in Southern Brazil.

Clinicopathologic Features of Tissue Granulomas in Transplant Recipients: A Single Center Study in a Nontuberculosis Endemic Region.

CONTEXT.­: There is a paucity of literature about tissue granulomas in transplant patients. OBJECTIVE.­: To characterize the clinicopathologic features of granulomas in this population and develop a clinically judicious approach to their evaluation. DESIGN.­: We performed chart reviews of solid organ and allogeneic hematopoietic stem cell transplant recipients at Yale New Haven Hospital to identify patients with granulomas on biopsy obtained pathologic specimens. Pretransplant and posttransplant specimens were included. Data points included demographics, clinical presentation, epidemiologic risk factors, biopsy indication, location and timing, immunosuppression, histopathology, microbiology, and associated clinical diagnosis. Granuloma-related readmissions and mortality were recorded at 1, 3, and 12 months. RESULTS.­: Biopsy proven granulomas were identified in 56 of 2139 (2.6%) patients. Of 56, 16 (29%) were infectious. Common infectious etiologies were bartonellosis (n = 3) and cytomegalovirus hepatitis (n = 3). Tuberculosis was not identified. Clinical symptoms prompted tissue biopsy in 27 of 56 (48.2%) cases while biopsies were obtained for evaluation of incidental findings or routine disease surveillance in 29 of 56 (51.8%). Presence of symptoms was significantly associated with infectious etiologies; 11 of 27 (40.7%) symptomatic patients compared with 5 of 29 (17.2%) asymptomatic patients had infectious causes. One death from granulomatous cryptogenic organizing pneumonia occurred. In pretransplant asymptomatic patients, no episodes of symptomatic disease occurred posttransplantation. CONCLUSIONS.­: Granulomas were uncommon in a large transplant population; most were noninfectious but presence of symptoms was associated with infectious etiologies. Granulomas discovered pretransplant without clear infectious etiology likely do not require prolonged surveillance after transplantation. Symptomatology and epidemiologic risks factors should guide extent of microbiologic evaluation.

Bartonella spp. detection in ticks, Culicoides biting midges and wild cervids from Norway.

Bartonella spp. are fastidious, gram-negative, aerobic, facultative intracellular bacteria that infect humans, and domestic and wild animals. In Norway, Bartonella spp. have been detected in cervids, mainly within the distribution area of the arthropod vector deer ked (Lipoptena cervi). We used PCR to survey the prevalence of Bartonella spp. in blood samples from 141 cervids living outside the deer ked distribution area (moose [Alces alces, n = 65], red deer [Cervus elaphus, n = 41] and reindeer [Rangifer tarandus, n = 35]), in 44 pool samples of sheep tick (Ixodes ricinus, 27 pools collected from 74 red deer and 17 from 45 moose) and in biting midges of the genus Culicoides (Diptera: Ceratopogonidae, 120 pools of 6,710 specimens). Bartonella DNA was amplified in moose (75.4%, 49/65) and in red deer (4.9%, 2/41) blood samples. All reindeer were negative. There were significant differences in Bartonella prevalence among the cervid species. Additionally, Bartonella was amplified in two of 17 tick pools collected from moose and in 3 of 120 biting midge pool samples. The Bartonella sequences amplified in moose, red deer and ticks were highly similar to B. bovis, previously identified in cervids. The sequence obtained from biting midges was only 81.7% similar to the closest Bartonella spp. We demonstrate that Bartonella is present in moose across Norway and present the first data on northern Norway specimens. The high prevalence of Bartonella infection suggests that moose could be the reservoir for this bacterium. This is the first report of bacteria from the Bartonella genus in ticks from Fennoscandia and in Culicoides biting midges worldwide.

Host conservation through their parasites: molecular surveillance of vector-borne microorganisms in bats using ectoparasitic bat flies.

Most vertebrates host a wide variety of haematophagous parasites, which may play an important role in the transmission of vector-borne microorganisms to hosts. Surveillance is usually performed by collecting blood and/or tissue samples from vertebrate hosts. There are multiple methods to obtain samples, which can be stored for decades if properly kept. However, blood sampling is considered an invasive method and may possibly be harmful to the sampled individual. In this study, we investigated the use of ectoparasites as a tool to acquire molecular information about the presence and diversity of infectious microorganism in host populations. We tested the presence of three distinct vector-borne microorganisms in both bat blood and bat flies: Bartonella bacteria, malaria-like Polychromophilus sp. (Apicomplexa), and Trypanosoma sp. (Kinetoplastea). We detected the presence of these microorganisms both in bats and in their bat flies, with the exception of Trypanosoma sp. in South African bat flies. Additionally, we found Bartonella sp. in bat flies from one population in Spain, suggesting its presence in the host population even if not detected in bats. Bartonella and Polychromophilus infection showed the highest prevalence in both bat and bat fly populations. Single, co- and triple infections were also frequently present in both. We highlight the use of haematophagous ectoparasites to study the presence of infectious microorganism in host blood and its use as an alternative, less invasive sampling method.

TITLE: Conservation des hôtes grâce à leurs parasites : surveillance moléculaire des microorganismes à transmission vectorielle chez les chauves-souris à l'aide de mouches ectoparasites. ABSTRACT: La plupart des vertébrés hébergent une grande variété de parasites hématophages, qui peuvent jouer un rôle important dans la transmission de microorganismes à transmission vectorielle à leurs hôtes. La surveillance est généralement effectuée en prélevant des échantillons de sang et/ou de tissus sur des hôtes vertébrés. Il existe plusieurs méthodes pour obtenir des échantillons, qui peuvent être conservés pendant des décennies dans des bonnes conditions. Cependant, le prélèvement sanguin est considéré comme une méthode invasive et peut éventuellement être nocif pour l'individu prélevé. Dans cette étude, nous avons étudié l'utilisation d'ectoparasites comme outil pour acquérir des informations moléculaires sur la présence et la diversité des microorganismes infectieux dans les populations hôtes. Nous avons testé la présence de trois microorganismes distincts, transmis par des vecteurs, dans le sang et les mouches des chauves-souris : les bactéries Bartonella, Polychromophilus sp. (Apicomplexa) et Trypanosoma sp. (Kinetoplastea). Nous avons détecté la présence de ces microorganismes à la fois chez les chauves-souris et chez leurs mouches des chauves-souris, à l'exception de Trypanosoma sp. chez les chauves-souris sud-africaines. De plus, nous avons trouvé Bartonella sp. chez les mouches des chauves-souris d'une population en Espagne, ce qui suggère sa présence dans la population hôte même si elle n'est pas détectée chez les chauves-souris elles-mêmes. Les infections à Bartonella et Polychromophilus ont montré la prévalence la plus élevée dans les populations de chauves-souris et de mouches des chauves-souris. Des infections simples, doubles et triples étaient également fréquemment présentes dans les deux cas. Nous mettons en évidence l'utilisation d'ectoparasites hématophages pour étudier la présence de microorganismes infectieux dans le sang de l'hôte et son utilisation comme méthode alternative et moins invasive d'échantillonnage.

Vector-Borne Pathogens in Ectoparasites Collected from High-Elevation Pika Populations.

The American pika, Ochotona princeps, is projected to decline throughout North America as climate change reduces its range, and pikas have already disappeared from several locations. In addition to climate, disease spillover from lower elevation mammalian species might affect pikas. We sampled pika fleas in Colorado and Montana across elevations ranging from 2896 to 3612 m and screened them for the presence of DNA from rodent-associated bacterial pathogens (Bartonella species and Yersinia pestis) to test the hypothesis that flea exchange between pikas and rodents may lead to occurrence of rodent-associated pathogens in pika ectoparasites. We collected 275 fleas from 74 individual pikas at 5 sites in Colorado and one site in Montana. We found that 5.5% of 275 pika fleas in this study tested positive for rodent-associated Bartonella DNA but that variation in Bartonella infection prevalence in fleas among sites was not driven by elevation. Specifically, we detected DNA sequences from two loci (gltA and rpoB) that are most similar to Bartonella grahamii isolates collected from rodents in Canada. We did not detect Y. pestis DNA in our survey. Our results demonstrate evidence of rodent-associated flea-borne bacteria in pika fleas. These findings are also consistent with the hypothesis that rodent-associated pathogens could be acquired by pikas. Flea-borne pathogen spillover from rodents to pikas has the potential to exacerbate the more direct effects of climate that have been suggested to drive pika declines.

Bartonella species and haplotypes in rodents and their fleas in Lanzarote and El Hierro in the Canary Islands, Spain.

Because isolated ecosystems contribute to species variability, especially oceanic island ecosystems, the present work focused on the study of the Bartonella species and haplotypes in Lanzarote and El Hierro, two Canary islands with evident bioclimatic differences between them. A total of 123 rodents and 110 fleas from two islands were screened for the presence of Bartonella by PCR analysis of the gltA and nuoG genes. The overall prevalence was 5.7% in rodents and 20.4% in fleas. A total of seven gltA-haplotypes was found in both rodents and fleas, belonging to the species Bartonella mastomydis and Bartonella tribocorum in Lanzarote, and to Bartonella rochalimae and Bartonella elizabethae in El Hierro, as well as recently described species Bartonella kosoyi in both islands. Besides, potential co-infections were detected based on the nuoG analysis. Further, Xenopsylla cheopis was the only flea species identified. Our study shows that isolated ecosystems such as the Canary Islands lead to the appearance of new Bartonella haplotypes along different biotopes, with diverse flea species involved in the spreading of the pathogen being of great relevance due to the zoonotic potential of the species found.

Detection of Anaplasma spp. and Bartonella spp. from wild-caught rodents and their ectoparasites in Nakhon Ratchasima Province, Thailand.

The objective of this study was to investigate evidence of emerging anaplasmosis and bartonellosis in rodents from endemic areas of Nakhon Ratchasima, Thailand. Rodent trapping was undertaken in 13 sub-districts of Muang District. The live-capture traps were set up in three locations of selected scrub typhus patient houses for three consecutive nights. Wild-caught rodent whole blood samples and associated ticks and fleas were collected and tested for Anaplasma spp. and Bartonella spp. In addition, heat maps using GIS software were used to determine the density of infection of positive wild-caught rodents. A total of 347 wild-caught rodents of nine species was captured. Rattus rattus (38.6%) was the dominant species. A total of 1,518 Heamaphysalis bandicota ticks and 57 Xenopsylla cheopis fleas was removed. Twenty-two of the 347 tested blood samples (6.3%) were Anaplasma bovis-positive and 121 blood samples and five out of 27 pools of X. cheopis fleas were Bartonella queenslandensis-positive. Of these infected rodents, dual-infections between A. bovis and B. queenslandensis were found in three B. indica rodents. Our results offer new information concerning the infections of A. bovis and B. queenslandensis in both rodents and their ectoparasites collected in high-risk areas of rodent-borne diseases in Thailand.

Molecular detection of Bartonella henselae, Bartonella clarridgeiae and Rickettsia felis in cat and dog fleas in Tenerife, Canary Islands, Spain.

The cat flea Ctenocephalides felis is the main vector of Bartonella henselae and Bartonella clarridgeiae, the causative agents of cat-scratch disease (CSD) and the spotted-fever agent Rickettsia felis. In spite of their worldwide distribution, there are no data on the occurrence of CSD-causing Bartonella species or the prevalence of Rickettsia species in the Canary Islands, Spain. Therefore, the aim of our study was to screen cat and dog fleas for both pathogens. A total of 128 C. felis from cats and dogs were screened for Bartonella and Rickettsia by PCR. Bartonella henselae (2.3%) and B. clarridgeiae (3.9%) were found in fleas infesting cats, whereas R. felis was identified in both cat (36.6%) and dog (40.7%) fleas. Further, co-infections were observed. This work constitutes the first finding of CSD-causing Bartonella species and the first study on the prevalence of R. felis in fleas from domestic animals in the Canary Islands. These results indicate public health importance, as associated infections could be misdiagnosed in the Archipelago despite their clinical relevance. Establishing human and animal routine diagnosis procedures for these pathogens along with improving vector control in shelters is necessary in order to prevent the spread of the infections among animals.

Detection and phylogenetic analysis of Bartonella species from bat flies on eastern bent-wing bats (Miniopterus fuliginosus) in Japan.

We examined Bartonella prevalence in 281 bat flies collected from 114 eastern bent-wing bats (Miniopterus fuliginosus) in Japan and phylogenetically analyzed with other bat fly and bat strains. The bat flies were identified as Penicilidia jenynsii (PJ; n = 45), Nycteribia allotopa (NA; n = 157), and novel Nycteribia species (NS; n = 79). Bartonella DNAs were detected in 31.7 % (89/281) of bat flies by PCR targeting the citrate synthase (gltA) gene. The prevalence of Bartonella DNA among the bat flies was 47.1 % (74/157) in NA, 15.2 % (12/79) in NS, and 6.7 % (3/45) in PJ. Bartonella bacteria were also isolated from two NA and one NS. A phylogenetic analysis of the gltA sequences revealed that bat fly-associated strains were classified into three lineages and the same lineages of Bartonella were commonly detected from both Nycteribia bat flies and Miniopterus bats. These results suggest that Nycteribia bat flies are potential vectors for transmitting Bartonella among Miniopterus bats.