First report of Bartonella sp. isolated from Hippobosca equina L. (Hippoboscidae: Hippobosca) in Lublin Province, south-eastern Poland.

INTRODUCTION AND OBJECTIVE: Hippobosca equina (Diptera: Hippoboscidae), is a widespread blood-feeding ectoparasite associated with the forest ecosystem. The insect is characterized by a wide host range and low host specificity, which increases the risk of feeding on animals that constitute a reservoir of transmissible pathogens, including Bartonella spp. MATERIAL AND METHODS: Hippobosca equina adults were collected from humans and companion animals within a continental mesotrophic oak-pine mixed forest in eastern Poland. DNA was isolated by the ammonia method, and isolates obtained from single individuals were tested by PCR method for the presence of 5 vector-borne pathogens. In case of the positive results, the amplicons were sequenced and examined by a BLAST search. RESULTS: The PCR analysis of DNA isolates obtained from 100 H. equina specimens revealed the presence of the RNA polymerase beta-subunit gene (rpoB) of the genus Bartonella, in 1% of the studied insects, i.e. one H. equina female. The rpoB gene haplotype of Bartonella sp. reported in this study, was identical to a Bartonella sp. sequence obtained from deer keds in Lithuania, and very closely related to strains with zoonotic potential. None of the H. equina specimens studied was positive for the presence of B. burgdorferi s.l., Anaplasma phagocytophilum, Babesia spp., and Coxiella burnetii. CONCLUSIONS: The study indicates the need to screen the occurrence of Bartonella spp., both in potential vectors and reservoirs of this pathogen in various habitats.

New records of pathogenic bacteria in different species of fleas collected from domestic and peridomestic animals in Spain. A potential zoonotic threat?

Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas.

Molecular detection of vector-borne agents in wild boars (Sus scrofa) and associated ticks from Brazil, with evidence of putative new genotypes of Ehrlichia, Anaplasma, and haemoplasmas.

The present study aimed to investigate, by molecular techniques, the occurrence of Anaplasmataceae, Bartonellaceae, Rickettsiaceae, Mycoplasmataceae, Coxiellaceae, and Babesiidae/Theileriidae agents in blood samples of free-living wild boars (Sus scrofa) and associated ticks in south-eastern Brazil. For this purpose, 67 blood samples and 265 ticks (264 Amblyomma sculptum and one Amblyomma ovale) were analysed. In the screening for Anaplasmataceae agents by a PCR assay based on the 16S rRNA gene, 5.97% blood samples and 50.54% ticks were positive. In the PCR assay for Ehrlichia spp. based on the dsb gene, 9.24% of ticks were positive. Despite the low occurrence, a possible new 16S rRNA genotype of Anaplasma sp. was detected in a wild boar's blood sample. According to phylogenetic analyses based on the 16S rRNA, gltA, and sodB genes and ITS (23S-5S rRNA) intergenic region, it was found that A. sculptum and A. ovale ticks collected from wild boars carry Ehrlichia genotypes phylogenetically associated with Ehrlichia ewingii, Ehrlichia ruminantium, and new Ehrlichia genotypes previously detected in horses, peccaries, and ticks collected from jaguars. In the screening for haemoplasmas by a qPCR based on the 16S rRNA gene, 88.06% of blood samples and 8.69% of ticks were positive. Mycoplasma suis, Mycoplasma parvum, and a possible new haemoplasma genotype were detected in wild boars in south-eastern Brazil. In the screening for Bartonella spp. using a nuoG-based qPCR assay, 3.8% of tick samples were positive. Phylogenetic inferences positioned four nuoG and one r gltA Bartonella sequences into the same clade as Bartonella machadoae. No blood or tick samples from wild boars showed to be positive in the qPCR for Coxiella burnetii based on the IS1111 gene. On the other hand, only 1.6% of ticks were positive in the nested PCR assay for piroplasmids based on the 18S rRNA gene. A 18S rRNA sequence detected in a pool of A. sculptum nymphs was phylogenetically close to Cytauxzoon felis sequences previously detected in cats from the United States. Rickettsia sp. closely related to Rickettsia bellii was detected in a pool of A. sculptum nymphs. This is the first report of haemoplasmas, B. machadoae, and Cytauxzoon spp. in A. sculptum. Wild boars and associated ticks do not seem to participate in the epidemiological cycle of C. burnetii in the region studied. This invasive mammal species may act as a potential disperser of ticks infected with Ehrlichia spp., Bartonella spp., haemotropic mycoplasmas, and Cytauxzoon, and may bring important epidemiological implications in the transmission of bartonelosis, ehrlichiosis, haemoplasmosis, and cytauxzoonosis to humans and animals, more specifically to horses, rodents, pigs, and cats.

Eco-epidemiological screening of multi-host wild rodent communities in the UK reveals pathogen strains of zoonotic interest.

Wild rodent communities represent ideal systems to study pathogens and parasites shared among sympatric species. Such studies are useful in the investigation of eco-epidemiological dynamics, improving disease management strategies and reducing zoonotic risk. The aim of this study was to investigate pathogen and parasites shared among rodent species (multi-host community) in West Wales in an area where human/wildlife disease risk was not previously assessed. West Wales is predominantly rural, with human settlements located alongside to grazing areas and semi-natural landscapes, creating a critical human-livestock-wildlife interface. Ground-dwelling wild rodent communities in Wales were live-trapped and biological samples - faeces and ectoparasites - collected and screened for a suite of pathogens and parasites that differ in types of transmission and ecology. Faecal samples were examined to detect Herpesvirus, Escherichia coli, and Mycobacterium microti. Ticks and fleas were collected, identified to species based on morphology and genetic barcodes, and then screened for Anaplasma phagocytophilum, Babesia microti, Borrelia burgdorferi sensu lato, and Bartonella sp. All the pathogens and parasites screened pose a characteristic epidemiological challenge, such as variable level of generalism, unknown zoonotic potential, and lack of data. The results showed that the bank vole Myodes glareolus had the highest prevalence of all pathogens and parasites. Higher flea species diversity was detected than in previous studies, and at least two Bartonella species were found circulating, one of which has not previously been detected in the UK. These key findings offer new insights into the distribution of selected pathogen and parasites and subsequent zoonotic risk, and provide new baselines and perspectives for further eco-epidemiological research.

First Detection of Bartonella spp. in Small Mammals from Rice Storage and Processing Facilities in Myanmar and Sri Lanka.

(1) Background: Bartonella spp. are zoonotic bacteria with small mammals as main reservoirs. Bartonella spp. prevalence in small mammals from Myanmar and Sri Lanka are yet unknown. (2) Methods: Small mammals were snap trapped in Sri Lanka and Myanmar in urban surroundings. Spleens-derived DNA was screened for Bartonella spp. using conventional PCR based on three target genes. Positive samples were sequenced. (3) Results: 994 small mammals were collected comprising 6 species: Bandicota bengalensis, Bandicota indica, Rattus exulans, Rattus rattus, Mus booduga, and Suncus murinus. In Myanmar, the Bartonella prevalence in Bandicoot rats (68.47%) was higher than in Rattus rattus (41.67%), Rattus exulans (21.33%), and Suncus murinus (3.64%). Furthermore the prevalence in Myanmar (34%, n = 495) was twice as high as in Sri Lanka (16%, n = 499). In Sri Lanka, Bartonella spp. occurred almost exclusively in R. rattus. In Myanmar, Bartonella kosoyi was mainly detected (56%), followed by Bartonella sp. KM2529 (15%), Bartonella sp. SE-Bart D (12%) and Bartonella henselae (1%). In Sri Lanka, B. phoceensis (60%) and Bartonella sp. KM2581 (33%) were predominant. (4) Conclusions: Bartonella spp. were detected in all investigated small mammal species from Myanmar and Sri Lanka for the first time. Bartonella kosoyi and B. henselae are zoonotic. As these small mammals originated from urban settlements, human bartonellosis seems likely to occur.

Bartonella infections diagnosed in the French reference center, 2014-2019, and focus on infections in the immunocompromised.

We studied retrospectively 651 PCR-confirmed Bartonella infections diagnosed at the French reference center for bartonellosis from 2014 to 2019. The most common form was cat-scratch disease (89%) followed by endocarditis (9%). Disseminated forms (2%) mainly presented as bacillary angiomatosis or peliosis hepatis in solid organ transplant recipients.

Laboratory Diagnosis of 37 Cases of Bartonella Endocarditis Based on Enzyme Immunoassay and Real-Time PCR.

Bartonella spp., mostly Bartonella quintana and B. henselae, are a common cause of culture-negative endocarditis. Serology using immunofluorescence assay (IFA) and PCR performed on cardiac tissues are the mainstays of diagnosis. We developed an enzyme immunoassay (EIA) and a novel multiplex real-time PCR assay, utilizing Bartonella genus-specific, B. henselae-specific, and B. quintana-specific SimpleProbe probes, for diagnosis of Bartonella endocarditis. We aimed to evaluate the performance of these assays. Thirty-seven patients with definite endocarditis, 18 with B. henselae, 18 with B. quintana, and 1 with B. koehlerae, were studied. Diagnosis was confirmed by conventional PCR and DNA sequencing of surgical cardiac specimens. Similar to the case with IFA, anti-Bartonella IgG titers of ≥1:800 were found in 94% of patients by EIA; cross-reactivity between B. henselae and B. quintana precluded species-specific serodiagnosis, and frequent (41%) but low-titer cross-reactivity between Coxiella burnetii antibodies and B. henselae antigen was found in patients with Q fever endocarditis. Low-titer (1:100) cross-reactivity was uncommonly found also in patients with brucellosis and culture-positive endocarditis, particularly Enterococcus faecalis endocarditis. Real-time PCR performed on explanted heart valves/vegetations was in complete agreement with results of sequence-based diagnosis with characteristic melting curves. The genus-specific probe identified five additional endocarditis-associated Bartonella spp. at the genus level. In conclusion, EIA coupled with a novel real-time PCR assay can play an important role in Bartonella endocarditis diagnosis and expand the diagnostic arsenal at the disposal of the clinical microbiologist. Since serology remains a major diagnostic tool, recognizing its pitfalls is essential to avoid incorrect diagnosis.

Prevalence of Bartonella sp. in United States military working dogs with infectious endocarditis: a retrospective case-control study.

OBJECTIVES: Bartonella infection has been associated with endocarditis in humans, dogs, cats and cattle. In order to evaluate the importance of this pathogen as a possible source of endocarditis in United States military working dogs (MWDs), we performed a retrospective case-control study on 26 dogs with histological diagnosis of culture negative endocarditis (n = 18), endomyocarditis (n = 5) or endocardiosis (n = 3) and 28 control dogs without any histological cardiac lesions. METHODS: DNA was extracted from paraffin embedded cardiac valves and tissues from case and control dogs and submitted to PCR testing with primers targeting the Bartonella gltA gene. PCR-RFLP using four restriction endonucleases and partial sequencing was then performed to determine the Bartonella species involved. RESULTS: Nineteen (73%) cases were PCR positive for Bartonella, including B. henselae (8 dogs), B. vinsonii subsp. berkhoffii (6 dogs), B. washoensis (2 dogs) and B. elizabethae (1 dog). Only one control dog was weakly PCR positive for Bartonella. Based on the type of histological diagnosis, 13 (72.2%) dogs with endocarditis, 3 (60%) dogs with endomyocarditis and all 3 dogs with endocardiosis were Bartonella PCR positive. CONCLUSIONS: Bartonella sp. Infections were correlated with cardiopathies in US military working dogs. Systemic use of insecticides against ectoparasites and regular testing of MWDs for Bartonella infection seem highly appropriate to prevent such life-threatening exposures.

Bartonella bovis and Candidatus Bartonella davousti in cattle from Senegal.

In Senegal, domestic ruminants play a vital role in the economy and agriculture and as a food source for people. Bartonellosis in animals is a neglected disease in the tropical regions, and little information is available about the occurrence of this disease in African ruminants. Human bartonellosis due to Bartonella quintana has been previously reported in Senegal. In this study, 199 domestic ruminants, including 104 cattle, 43 sheep, and 52 goats were sampled in villages from the Senegalese regions of Sine Saloum and Casamance. We isolated 29 Bartonella strains, all exclusively from cattle. Molecular and genetic characterization of isolated strains identified 27 strains as Bartonella bovis and two strains as potentially new species. The strains described here represent the first Bartonella strains isolated from domestic ruminants in Senegal and the first putative new Bartonella sp. isolated from cattle in Africa.

Update on blood culture-negative endocarditis.

Blood culture-negative endocarditis is often severe, and difficult to diagnose. The rate of non-documented infective endocarditis has decreased with the advent of molecular biology - improved performance for the diagnosis of bacterial endocarditis with blood cultures sterilized by previous antibacterial treatment - and cardiac surgery - access to the main infected focus, the endocardium, for half of the patients. Blood culture-negative endocarditis are classified in 3 main categories: (i) bacterial endocarditis with blood cultures sterilized by previous antibacterial treatment (usually due to usual endocarditis-causing bacteria, i.e. streptococci, more rarely staphylococci, or enterococci); (ii) endocarditis related to fastidious microorganisms (e.g. HACEK bacteria; defective streptococci - Gemella, Granulicatella, and Abiotrophia sp. - Propionibacterium acnes, Candida sp.): in these cases, prolonged incubation will allow identifying the causative pathogen in a few days; (iii) and the "true" blood culture-negative endocarditis, due to intra-cellular bacteria that cannot be routinely cultured in blood with currently available techniques: in France, these are most frequently Bartonella sp., Coxiella burnetti (both easily diagnosed by ad hoc serological tests), and Tropheryma whipplei (usually diagnosed by PCR on excised cardiac valve tissue). Non-infective endocarditis is rare, mostly limited to marantic endocarditis, and the rare endocarditis related to systemic diseases (lupus, Behçet).

Molecular detection of feline arthropod-borne pathogens in cats in Cuiabá, state of Mato Grosso, central-western region of Brazil / Detecção molecular de patógenos transmitidos por artrópodes em gatos de Cuiabá, estado do Mato Grosso, centro-oeste do Brasil

Hemotrophic mycoplasmas (hemoplasmas), Bartonellasp., Hepatozoon sp. and Cytauxzoon felis are prominent pathogens that circulate between cats and invertebrate hosts. The present study aimed to detect the presence of DNA from hemoplasmas, Bartonella sp., Hepatozoon sp. and Cytauxzoon felis, and then confirm it by means of sequencing, in blood samples from cats in Cuiabá, MT, Brazil. From February 2009 to February 2011, blood samples with added EDTA were collected from 163 cats that were being housed in four different animal shelters in the city of Cuiabá, state of Mato Grosso, Brazil and from 15 cats that were admitted to the veterinary hospital of the Federal University of Mato Grosso (UFMT). Out of the 178 cats sampled, 15 (8.4%) were positive for hemoplasmas: four (2.2%) for Mycoplasma haemofelis, 12 (6.7%) for 'Candidatus M. haemominutum' and one (0.5%) for 'Candidatus M. turicensis'. One cat (0.5%), a patient that was attended at the veterinary hospital, was coinfected with M. haemofelis, 'Candidatus M. haemominutum' and 'Candidatus M. turicensis', based on sequencing confirmation. Four cats were positive for Bartonella spp.: three (1.7%) for B. henselae and one (0.5%) for B. clarridgeiae. None of the animals showed Cytauxzoon sp. or Hepatozoon sp. DNA in their blood samples. This study showed that cats housed in animal shelters in the city of Cuiabá, state of Mato Grosso, are exposed to hemoplasmas and Bartonella species.

Micoplasmas hemotróficos (hemoplasmas), Bartonellasp., Hepatozoon sp. e Cytauxzoon felis se destacam como importantes patógenos que circulam entre gatos e hospedeiros invertebrados. O presente estudo objetivou detectar e, posteriormente confirmar por seqüenciamento, a presença de DNA de hemoplasmas, Bartonella sp., Hepatozoon sp. e Cytauxzoon felis em amostras de sangue de gatos de Cuiabá, MT, Brasil. Entre fevereiro/2009 e fevereiro de 2011, amostras de sangue acrescidas de EDTA foram coletadas de 163 gatos mantidos em quatro diferentes abrigos na cidade de Cuiabá, estado do Mato Grosso, Brasil, e de 15 gatos atendidos no Hospital Veterinário da Universidade Federal do Mato Grosso (UFTM). Dos 178 gatos amostrados, 15 (8,4%) foram positivos para hemoplasmas: quatro (2,2%) para Mycoplasma haemofelis, 12 (6,7%) para 'Candidatus M. haemominutum' e um (0,5%) para 'Candidatus M. turicensis'. Um (0.5%) gato, atendido no Hospital Veterinário da UFMT, estava co-infectado com M. haemofelis, 'Candidatus M. haemominutum' e 'Candidatus M. turicensis', baseado na confirmação por sequenciamento. Quatro gatos mostraram-se positivos para Bartonella spp.: três (1,7%) para B. henselae e um (0.5%) para B. clarridgeiae. Todos os gatos amostrados mostraram-se negativos para Cytauxzoon sp. e Hepatozoon sp. Este estudo mostrou que gatos mantidos em abrigos na cidade de Cuiabá, estado do Mato Grosso, são expostos a hemoplasmas e espécies de Bartonella sp.