Bartonella Henselae Research Articles

Occurrence and bacterial loads of Bartonella and haemotropic Mycoplasma species in privately owned cats and dogs and their fleas from East and Southeast Asia

Bartonella spp. and haemoplasmas are pathogens of veterinary and medical interest with ectoparasites mainly involved in their transmission. This study aimed at molecular detection of Bartonella spp. and haemoplasmas in cats (n = 93) and dogs (n = 96), and their related fleas (n = 189) from countries in East and Southeast Asia. Ctenocephalides felis was the dominant flea species infesting both cats (97.85%) and dogs (75%) followed by Ctenocephalides orientis in dogs (18.75%) and rarely in cats (5.2%). Bartonella spp. DNA was only detected in blood samples of flea‐infested cats (21.51%) (p < .0001, OR = 27.70) with Bartonella henselae more frequently detected than Bartonella clarridgeiae in cat hosts (15.05%, 6.45%) and their associated fleas (17.24%, 13.79%). Out of three Bartonella‐positive fleas from dogs, two Ct. orientis fleas carried Bartonella vinsonii subsp. berkhoffii and Bartonella clarridgeiae, while the 3rd flea (Ct. felis) carried Candidatus Bartonella merieuxii. Felines represented a risk factor for Bartonella spp. infections, where fleas collected from cats (32.25%) presented an increased likelihood for Bartonella spp. occurrence (p < .0001, OR = 14.76) than those from dogs (3.13%). Moreover, when analysing infectious status, higher Bartonella spp. DNA loads were detected in fleas from bacteraemic cats compared to those from non‐bacteraemic ones (p < .05). The haemoplasma occurrence was 16.13% (15/93) and 4.17% (4/96) in cat and dog blood samples from different countries (i.e. Indonesia, Malaysia, the Philippines, Taiwan and Thailand), with cats more at risk of infection (p < .01, OR = 5.96) than dogs. Unlike Bartonella spp., there was no evidence for flea involvement in the hemoplasmas' transmission cycle, thus supporting the hypothesis of non‐vectorial transmission for these pathogens. In conclusion, client‐owned cats and dogs living in East and Southeast Asia countries are exposed to vector‐borne pathogens with fleas from cats playing a key role in Bartonella spp. transmission, thus posing a high risk of infection for humans sharing the same environment.

Bartonella Endocarditis in Spain: Case Reports of 21 Cases

Blood culture negative endocarditis (BCNE) is frequent in infective endocarditis (IE). One of the causes of BCNE is fastidious microorganisms, such as Bartonella spp. The aim of this study was to describe the epidemiologic, clinical characteristics, management and outcomes of patients with Bartonella IE from the “Spanish Collaboration on Endocarditis-Grupo de Apoyo al Manejo de la Endocarditis infecciosa en España (GAMES)”cohort. Here we presented 21 cases of Bartonella IE. This represents 0.3% of a total of 5590 cases and 2% of the BCNE from the GAMES cohort. 62% were due to Bartonella henselae and 38% to Bartonella quintana. Cardiac failure was the main presenting form (61.5% in B. hensalae, 87.5% in B. quintana IE) and the aortic valve was affected in 85% of the cases (76% in B. henselae, 100% in B. quintana IE). Typical signs such as fever were recorded in less than 40% of patients. Echocardiography showed vegetations in 92% and 100% of the patients with B. henselae and B. quintana, respectively. Culture was positive only in one patient and the remaining were diagnosed by serology and PCR. PCR was the most useful tool allowing for diagnosis in 16 patients (100% of the studied valves). Serology, at titers recommended by guidelines, only coincided with PCR in 52.4%. Antimicrobial therapy, in different combinations, was used in all cases. Surgery was performed in 76% of the patients. No in-hospital mortality was observed. One-year mortality was 9.4%. This article remarks the importance for investigating the presence of Bartonella infection as causative agent in all BCNE since the diagnosis needs specific microbiological tools and patients could benefit of a specific treatment.

Choroidal granulomas due to Bartonella henselae infection: A case series

To report a case series of 3 patients with choroidal granulomas due to Bartonella henselae infection in order to raise awareness about this etiology in the differential diagnosis of choroidal granulomas. A retrospective case series of patients with choroidal granulomas due to Bartonella henselae infection who consulted between 2018 and 2020. Data were collected from the medical records (demographics, visual acuity (VA), laboratory tests, treatment, imaging). Patients were a 48-year old man, a 14-year old girl and a 31-year old man. They all had a choroidal granuloma seen on optical coherence tomography (OCT) and angiography. The laboratory work-up revealed a positive serology for Bartonella henselae in all patients. On multimodal imaging choroidal granulomas in B Henselae appeared as single or multiple, uni or bilateral round yellowish lesions. Fluorescein and indocyanine green angiography of the granuloma showed respectively a late staining and a hypofluorescence. On EDI-OCT choroidal granuloma appeared as a round hyporeflective lesion in the choroid with a retinal elevation. The exclusion of other diagnosis, the natural course and the serology must lead the ophthalmologist to evoke the diagnosis.

Serological and Molecular Detection of Bartonella henselae in Cats and Humans From Egypt: Current Status and Zoonotic Implications

Bartonellosis is a vector-borne zoonotic disease caused by the intracellular bacterium of genus Bartonella. The disease has a worldwide distribution and cats represent the major reservoir of this disease. Despite its global distribution, very limited previous studies have investigated the occurrence of bartonellosis in cats and their owners in Egypt. In an endeavor to explore this topic, we investigated the occurrence of Bartonella henselae (B. henselae) infection in 225 samples (blood, saliva, and claw) obtained from 75 healthy cats in Upper Egypt. These samples were routinely obtained during veterinary clinic visits. This study also involved an examination of 100 humans, including cat owners and people with a history of contact with cats. Attempted isolation and identification of B. henselae in cats were also performed. Furthermore, PCR was performed for molecular identification of B. henselae in blood samples from cats. Meanwhile, an immunofluorescent assay was performed to study the seroprevalence of B. henselae infection in humans. In this study, B. henselae could not be isolated from any of the examined blood, saliva, or claw samples from cats. Interestingly, B. henselae was identified molecularly in 8% (6/75) of blood samples from cats. The seroprevalence of B. henselae in humans was 46% and its occurrence was higher in females (46.6%) than in males (41.7%) (P = 0.748). B. henselae infection was higher among cat owners [51.4% (19/37)] than among people with a history of contact with cats [42.9% (27/63)] (P = 0.410). Infection was higher in rural regions [79.5% (31/39)] than in urban regions [24.6% (15/61)] (P < 0.001). Collectively, this data provide interesting baseline information about the occurrence of B. henselae in cats and humans in Upper Egypt, which reflects the potential zoonotic transmission of this bacterium. Future study is mandatory to explore the occurrence of B. henselae in major reservoirs in Egypt.

Child Neurology: Bartonella henselae Neuroretinitis in 2 Patients.

Neuroretinitis due to Bartonella henselae infection is a rare cause of vision loss in children. Two pediatric cases of acute unilateral vision loss accompanied by edema of the optic nerve on fundoscopic examination are presented. Severe causes of vision loss were excluded. During the course of the disease, macular stellate exudates emerged on control fundoscopic examinations, and diagnosis of neuroretinitis was made. A causative agent was confirmed by serologic examination, as high titers of IgM and IgG antibodies to Bartonella henselae were detected. Both patients significantly recovered after oral antibiotic treatment.

Molecular Diagnosis of Cat Scratch Disease: a 25-Year Retrospective Comparative Analysis of Various Clinical Specimens and Different PCR Assays.

ABSTRACTCat-scratch disease (CSD), caused primarily by Bartonella henselae, is a common etiology of infectious regional lymphadenopathy. Lymphadenopathy is preceded by a primary inoculation lesion and may progress to suppuration. Laboratory diagnosis of CSD is hampered by the limitations of available confirmatory tests. PCR, in general, is highly sensitive and specific; however, clinical sensitivity in CSD varies greatly between studies. We aimed to identify clinical specimens and PCR assays best suited for CSD diagnosis using a national CSD registry and a uniform case definition. Different clinical specimens and PCR assays, including conventional and real-time PCR, were evaluated. PCR was positive in 335/390 (86%) CSD patients and 425/482 (88%) PCR tests. The highest PCR sensitivity was achieved in lymph node pus aspirates (96%; n = 278 tests) followed by primary lesions (88%; n = 50), lymph node fine needle aspirations (85%; n = 46), lymph node biopsy specimens (73%; n = 91) and paraffin-embedded lymph nodes (59%; n = 17), (P < 0.001). Sensitivity was similar in all types of PCR assays studied. PCR negative predictive value of pus aspirate and lymph node biopsy specimen patient groups was 82% and 72%, respectively. Specificity was 100% based on 125 non-CSD patients with negative PCR. In conclusion, the specimen type rather than the PCR assay type has a major impact on CSD molecular diagnosis. We assume that the inadequate sensitivity of the biopsy specimens was due to sampling errors or the presence of inhibitory factors. Primary lesions should be sampled more frequently for CSD diagnosis. Physicians should be aware of the low PCR negative predictive value of lymph node biopsy specimens.IMPORTANCE Polymerase chain reaction (PCR) for the detection of Bartonella henselae is an important tool for the diagnosis of cat scratch disease (CSD); however, clinical sensitivity varies greatly between studies. The current study shows that the specimen type, with pus aspiration, fine needle aspiration, and primary inoculation lesion having significantly higher sensitivity than fresh or formalin-fixed paraffin-embedded lymph node biopsy specimen, rather than the type of the PCR assay, whether a conventional or a real-time assay, has a major impact on the performance of diagnostic PCR for CSD. The new data provide new tools for the clinical microbiologist when interpreting the results of the PCR assays. Primary inoculation lesions, although easily accessible, are often neglected and should be sampled more frequently for molecular diagnosis of CSD. Physicians should be aware that negative PCR, particularly if performed on fresh or paraffin-embedded lymph node biopsy specimens, does not exclude CSD.

Molecular screening of cat and dog ectoparasites for the presence of Bartonella spp. in Attica, Greece.

The purpose of this study was the molecular detection of Bartonella spp. in fleas and ticks parasitizing cats and dogs from 39 locations in Attica, Greece. One hundred and forty five ectoparasites (104 fleas and 41 ticks) from 92 cats and 53 dogs were investigated individually using PCRs targeting the 16S-23S ribosomal RNA intergenic spacer (ITS) and the citrate synthase (gltA) genetic loci. Bartonella spp. were detected in 14 out of 104 fleas (13.5%) and in none of the ticks examined. Consequent sequence analysis of the amplicons from the two loci identified 3 strains as Bartonella henselae, and 11 as Bartonella clarridgeiae. Οur study demonstrates the presence of B. henselae and B. clarridgeiae in Ctenocephalides felis fleas from cat and dog in Greece. We also report a novel ITS sequence for B. clarridgeiae. Considering that fleas could pose a risk for human bartonellosis from their infected hosts, further studies on the public health risk of Bartonella presence in animal ectoparasites are warranted.

A Pain in the Neck: An Adolescent with Neck Pain.

A Pain in the Neck: An Adolescent with Neck Pain.

Molecular detection of Bartonella henselae in blood samples obtained from owned cats in Kerman city using Nested-PCR

Molecular detection of Bartonella henselae in blood samples obtained from owned cats in Kerman city using Nested-PCR

Overexpression of a "Candidatus Liberibacter Asiaticus" Effector Gene CaLasSDE115 Contributes to Early Colonization in Citrus sinensis.

Huanglongbing (HLB), caused by “Candidatus liberibacter asiaticus” (CaLas), is one of the most devastating diseases in citrus but its pathogenesis remains poorly understood. Here, we reported the role of the CaLasSDE115 (CLIBASIA_05115) effector, encoded by CaLas, during pathogen-host interactions. Bioinformatics analyses showed that CaLasSDE115 was 100% conserved in all reported CaLas strains but had sequence differences compared with orthologs from other “Candidatus liberibacter.” Prediction of protein structures suggested that the crystal structure of CaLasSDE115 was very close to that of the invasion-related protein B (IalB), a virulence factor from Bartonella henselae. Alkaline phosphatase (PhoA) assay in E. coli further confirmed that CaLasSDE115 was a Sec-dependent secretory protein while subcellular localization analyses in tobacco showed that the mature protein of SDE115 (mSDE115), without its putative Sec-dependent signal peptide, was distributed in the cytoplasm and the nucleus. Expression levels of CaLasSDE115 in CaLas-infected Asian citrus psyllid (ACP) were much higher (∼45-fold) than those in CaLas-infected Wanjincheng oranges, with the expression in symptomatic leaves being significantly higher than that in asymptomatic ones. Additionally, the overexpression of mSDE115 favored CaLas proliferation during the early stages (2 months) of infection while promoting the development of symptoms. Hormone content and gene expression analysis of transgenic plants also suggested that overexpressing mSDE115 modulated the transcriptional regulation of genes involved in systemic acquired resistance (SAR) response. Overall, our data indicated that CaLasSDE115 effector contributed to the early colonization of citrus by the pathogen and worsened the occurrence of Huanglongbing symptoms, thereby providing a theoretical basis for further exploring the pathogenic mechanisms of Huanglongbing disease in citrus.

Érythème noueux compliquant une maladie des griffes du chat : une association et des implications thérapeutiques à connaître

IntroductionCat-scratch disease is a zoonosis caused by Bartonella henselae and transmitted mostly through a break in the skin. This infectious disease is commonly characterized by a benign lymphadenitis. However, more unusual presentations have been observed. Case reportWe report the case of a 44-year-old woman with cat-scratch disease who developed subsequently an erythema nodosum. The non-steroidal anti-inflammatory treatment of the erythema nodosum may have led to worsening of cat-scratch disease with an abscess requiring surgical treatment. ConclusionCat-scratch disease is an uncommon and poorly known cause of erythema nodosum, and must be systematically ruled out. As non-steroidal anti-inflammatory drug are usually prescribed as a treatment of erythema nodosum, it should be used cautiously to reduce the risk of complications in the cat-scratch disease.

Long-Read Sequencing Reveals Genetic Adaptation of Bartonella Adhesin A Among Different Bartonella henselae Isolates.

Bartonella henselae is the causative agent of cat scratch disease and other clinical entities such as endocarditis and bacillary angiomatosis. The life cycle of this pathogen, with alternating host conditions, drives evolutionary and host-specific adaptations. Human, feline, and laboratory adapted B. henselae isolates often display genomic and phenotypic differences that are related to the expression of outer membrane proteins, for example the Bartonella adhesin A (BadA). This modularly-structured trimeric autotransporter adhesin is a major virulence factor of B. henselae and is crucial for the initial binding to the host via the extracellular matrix proteins fibronectin and collagen. By using next-generation long-read sequencing we demonstrate a conserved genome among eight B. henselae isolates and identify a variable genomic badA island with a diversified and highly repetitive badA gene flanked by badA pseudogenes. Two of the eight tested B. henselae strains lack BadA expression because of frameshift mutations. We suggest that active recombination mechanisms, possibly via phase variation (i.e., slipped-strand mispairing and site-specific recombination) within the repetitive badA island facilitate reshuffling of homologous domain arrays. The resulting variations among the different BadA proteins might contribute to host immune evasion and enhance long-term and efficient colonisation in the differing host environments. Considering the role of BadA as a key virulence factor, it remains important to check consistently and regularly for BadA surface expression during experimental infection procedures.

Serious and Atypical Presentations of Bartonella henselae Infection in Kidney Transplant Recipients.

This article describes 5 cases of bartonellosis with fever and atypical clinical presentations in kidney transplant recipients: thrombotic microangiopathies, recurrent hemophagocytosis, and immune reconstitution syndrome after treatment. The diagnosis, the pathological lesions, and treatments are described. Bartonellosis must be researched in solid organ transplant recipients with fever of undetermined origin.

Infections ostéo-articulaires à bactéries rares ou atypiques

Microbiological identification is essential for the management of bone and joint infections. However, the detection of the pathogen is sometimes difficult. In case of a negative sample and/or depending on the context, the presence of so-called atypical bacteria is to be mentioned and looked for. This article reports the main characteristics of osteoarticular infections caused by atypical mycobacteria, Brucella sp., Pasteurella sp., Bartonella henselae and Kingella kingae. Tuberculosis infections and Whipple's disease will be discussed in other articles in the journal.

Using Proteomic Approaches to Unravel the Response of Ctenocephalides felis felis to Blood Feeding and Infection With Bartonella henselae.

Among the Ctenocephalides felis felis-borne pathogens, Bartonella henselae, the main aetiological agent of cat scratch disease (CSD), is of increasing comparative biomedical importance. Despite the importance of B. henselae as an emergent pathogen, prevention of the diseases caused by this agent in cats, dogs and humans mostly relies on the use of ectoparasiticides. A vaccine targeting both flea fitness and pathogen competence is an attractive choice requiring the identification of flea proteins/metabolites with a dual effect. Even though recent developments in vector and pathogen -omics have advanced the understanding of the genetic factors and molecular pathways involved at the tick-pathogen interface, leading to discovery of candidate protective antigens, only a few studies have focused on the interaction between fleas and flea-borne pathogens. Taking into account the period of time needed for B. henselae replication in flea digestive tract, the present study investigated flea-differentially abundant proteins (FDAP) in unfed fleas, fleas fed on uninfected cats, and fleas fed on B. henselae-infected cats at 24 hours and 9 days after the beginning of blood feeding. Proteomics approaches were designed and implemented to interrogate differentially expressed proteins, so as to gain a better understanding of proteomic changes associated with the initial B. henselae transmission period (24 hour timepoint) and a subsequent time point 9 days after blood ingestion and flea infection. As a result, serine proteases, ribosomal proteins, proteasome subunit α-type, juvenile hormone epoxide hydrolase 1, vitellogenin C, allantoinase, phosphoenolpyruvate carboxykinase, succinic semialdehyde dehydrogenase, glycinamide ribotide transformylase, secreted salivary acid phosphatase had high abundance in response of C. felis blood feeding and/or infection by B. henselae. In contrast, high abundance of serpin-1, arginine kinase, ribosomal proteins, peritrophin-like protein, and FS-H/FSI antigen family member 3 was strongly associated with unfed cat fleas. Findings from this study provide insights into proteomic response of cat fleas to B. henselae infected and uninfected blood meal, as well as C. felis response to invading B. henselae over an infection time course, thus helping understand the complex interactions between cat fleas and B. henselae at protein levels.

Misleading Positive Serology for Cat Scratch Disease following Administration of Intravenous Immunoglobulin.

Cat scratch disease (CSD), caused by Bartonella henselae, usually presents as regional lymphadenopathy/lymphadenitis, known as typical CSD or as atypical CSD, which includes, among others, neurological manifestations. Serology for anti-B. henselae IgG antibodies is the most commonly used diagnostic tests for CSD. Intravenous immunoglobulin (IVIG) is given for an increasing number of medical conditions and may cause interference with serological testing. We report six patients with neurological manifestations and two patients with Kawasaki disease mimicking typical CSD, mistakenly diagnosed as CSD due to false-positive serology following IVIG therapy. Bartonella IgG serology was positive one to six days after IVIG administration and reverted to negative in seven of eight patients or significantly decreased (1 patient) ≤30 days later. In patients with CSD, IgG titers remained essentially unchanged 15–78 days after the positive serum sample. An additional eight patients treated with IVIG for various conditions were evaluated prospectively. All were seronegative one day pre-IVIG infusion, five patients demonstrated an increase in the IgG titers one to three days after IVIG administration, one interpreted as positive and four as intermediate, whereas three patients remained seronegative, suggesting that false seropositivity after IVIG therapy may not occur in all patients. Treatment with IVIG can result in false-positive serology for B. henselae. Increased awareness to the misleading impact of IVIG is warranted to avoid misinterpretation. Repeat testing can distinguish between true and false serology. Preserving serum samples prior to IVIG administration is suggested.

Prospective serological and molecular cross-sectional study focusing on Bartonella and other blood-borne organisms in cats from Catalonia (Spain)

BackgroundThere is limited clinical or epidemiological knowledge regarding Bartonella infection in cats, and no serological studies have compared the presence of antibodies against different Bartonella species. Moreover, there are limited feline Bartonella studies investigating co-infections with other vector-borne pathogens and the associated risk factors. Therefore, the objective of this study was to investigate Bartonella spp. infections and co-infections with other pathogens in cats from Barcelona (Spain) based on serological and/or molecular techniques and to determine associated risk factors.MethodsWe studied colony and owned cats (n = 135). Sera were tested for Bartonella henselae-, Bartonella quintana-, and Bartonella koehlerae-specific antibodies using endpoint in-house immunofluorescence antibody assays. Bartonella real-time PCR (qPCR) and conventional PCR (cPCR) were performed. In addition, cPCR followed by DNA sequencing was performed for other pathogenic organisms (Anaplasma, Babesia, Cytauxzoon, Ehrlichia, Hepatozoon, hemotropic Mycoplasma, and Theileria spp.).ResultsFrom 135 cats studied, 80.7% were seroreactive against at least one Bartonella species. Bartonella quintana, B. koehlerae, and B. henselae seroreactivity was 67.4, 77.0, and 80.7%, respectively. Substantial to almost perfect serological agreement was found between the three Bartonella species. Colony cats were more likely to be Bartonella spp.-seroreactive than owned cats. Moreover, cats aged ≤ 2 years were more likely to be Bartonella spp.-seroreactive. Bartonella spp. DNA was detected in the blood of 11.9% (n = 16) of cats. Cats were infected with B. henselae (n = 12), B. clarridgeiae (n = 3), and B. koehlerae (n = 1). Mycoplasma spp. DNA was amplified from 14% (n = 19) of cat blood specimens. Cats were infected with Mycoplasma haemofelis (n = 8), Candidatus M. haemominutum (n = 6), Candidatus Mycoplasma turicensis (n = 4), and Mycoplasma wenyonii (n = 1). Anaplasma, Babesia, Cytauxzoon, Ehrlichia spp., Hepatozoon, and Theileria spp. DNA was not amplified from any blood sample. Of the 16 Bartonella spp.-infected cats based on PCR results, six (37%) were co-infected with Mycoplasma spp.ConclusionsBartonella spp. and hemoplasma infections are prevalent in cats from the Barcelona area, whereas infection with Anaplasma spp., Babesia, Cytauxzoon, Ehrlichia spp., Hepatozoon, and Theileria infections were not detected. Co-infection with hemotropic Mycoplasma appears to be common in Bartonella-infected cats. To our knowledge, this study is the first to document M. wenyonii is infection in cats.Graphical

Atlantoaxial instability secondary to Bartonella henselae osteomyelitis managed surgically by atlantoaxial instrumentation: A case report and systematic review.

Cat scratch disease (CSD), caused by Bartonella henselae, may atypically present with vertebral osteomyelitis. Antibiotic regimens are tailored to presentation, which is markedly variable and not well defined for any atypical disease. In cases of spinal instability, the use of antibiotics alone may not be sufficient. Atlantoaxial instability caused by osteomyelitis is a rare complication of CSD. In this report, we describe the rare case of vertebral osteomyelitis complicated by atlantoaxial instability, requiring both antibiotics and atlantoaxial fusion. We discuss our case, surgical technique, rationale, and outcome. In addition, we conducted a systematic review of the literature of vertebral osteomyelitis in pediatric secondary to B. henselae. A 2-year-old child presented with a 2-month history of irritability, fever, and rigid neck pain along with a recent history of feline exposure. Physical examination revealed cervical tenderness and decreased range of motion. Computed tomography (CT) showed osteolysis of the right C1 lateral mass and pars articularis; T1-weighted magnetic resonance imaging with contrast showed enhancement around the right C1 lateral mass. The titer for B. henselae was high. A diagnosis of cat scratch osteomyelitis with cervical instability was made, for which the patient underwent surgery with atlantoaxial fusion. Postoperative imaging demonstrated resolution of the contrast-enhanced lesion. At 6-year follow-up, the patient showed no signs of residual complications from surgical intervention with a solid fusion. Our review revealed 44 cases of pediatric CSD vertebral osteomyelitis. Conservative management with antibiotic employed in 86% while antibiotics with surgical intervention in 14% of the cases. Surgical intervention was most often in the form of incision for drainage and decompression without fusion. Average follow-up 10 months with 86% achieved complete resolution. Cervical instability caused by osteolysis is a rare complication of CSD. This can subsequently lead to vertebral instability, requiring definitive surgical intervention.

Molecular detection and characterization of the endosymbiont Wolbachia in the European hedgehog flea, Archaeopsylla erinacei.

Wolbachia, the endosymbiont of arthropods and onchocercid nematodes is present in many medically important insect species, being also considered for the indirect control of parasitic ones. Archaeopsylla erinacei is a flea species infesting hedgehogs acting as vector of Rickettsia felis, Bartonella henselae, and Rickettsia helvetica, thus having public health relevance. The Wolbachia surface protein (wsp) and 16S rRNA genes were used to determine the presence, prevalence and molecular typing of Wolbachia in this flea species collected in two regions of southern Italy. Of the 45 fleas tested (n=16 males, 35.6%; n=29 females, 64.4%), 43 (95.6%; 95% CI: 84.8-99.2) scored positive for Wolbachia, of which 15 (33.3%) and 28 (62.2%) were males and females, respectively. The sex-wise prevalence of this endosymbiont was almost equal in both sexes (males 93.8%; 95% CI: 69.5-99.7; females 96.7%; 95% CI: 83.1-99.8). Single locus sequence analysis (SLST) of Wolbachia revealed two sequence types for 16S rRNA gene, named as wAr_15227 and wAr_15234, which came from two different areas, equally distributed in male and female fleas, whilst only one sequence type was identified for wsp gene. The phylogenetic analysis placed the two 16S rRNA sequence types in paraphyletic clades belonging to the supergroup A and B, respectively. Whilst, the tree of wsp gene clustered the corresponding sequence in the same clade including those of Wolbachia supergroup A. In MLST analyses, both Wolbachia sequence types clustered in a monophyletic clade with Drosophila nikananu (wNik) and Drosophila sturtevanti (wStv) from supergroup A. ClonalFrame analysis revealed a recombination event in the wAr_15234 strain which came from Apulia region. Scientific knowledge of the presence/prevalence of Wolbachia among medically important fleas, may contribute to develop an alternative biological method for the vector control.

A review of public health important fleas (Insecta, Siphonaptera) and flea-borne diseases in India.

Fleas (Insecta, Siphonaptera) are important vectors of plague and murine typhus in many parts of the world. Currently, about 2700 flea species were described in the world. The most common vector flea Xenopsylla cheopis is found throughout India, but X. astia, and X. brasiliensis are found less and limited in distribution associated with the domestic rats such as Rattus rattus, R. norvegicus, Mus musculus, and Bandicota bengalensis. Bubonic plague is a major flea-borne disease caused by the bacterial pathogen Yersinia pestis, transmitted from rats to humans via the rodent flea, X. cheopis. A major outbreak of plague and high mortality occurred in India. After 1966 with the 3 decadal intervals, plague cases occurred only during the year 1994 reported in 5 different states (Gujarat, Maharashtra, Karnataka, Uttar Pradesh, Madhya Pradesh and New Delhi and subsequently plague cases occurred during 2002 and 2004 after the one-decade interval in Himachal Pradesh (2002). Another outbreak of bubonic plague was reported in Dangud village, Barkhot tehsil, Uttarkashi district, Uttarakhand during October 2004. Ctenocephalides fleas are common in cats and dogs, which are the main vectors of bacteria rickettsiae, such as Rickettsia typhi, R. felis, R. conorii, and Bartonella henselae. Molecular and serological evidence also confirms the presence of R. typhi, R. conorii R. felis and B. henselae pathogens in cats and other fleas in India. Flea bites and flea-borne dermatitis are common in men and pet animals. Because of the re-emergence of the plague, updated information on fleas and flea-borne diseases are essential to control the flea vectors and flea-borne diseases in India. Hence, this comprehensive review updates the available information on fleas and fleas transmitted diseases in India.