Serological and Molecular Investigations of Babesia Microti in Dogs from Southern Italy

Piroplasmosis is now considered emerging tick-borne zoonosis worldwide and domestic animals have been proposed as potential reservoirs for some piroplasm infections. The aim of this research was to identify the frequency of Babesia microti infection in untravelled dogs from Southern Italy (Sicily). Blood samples from 89 dogs were examined for the presence of Babesia microti antibodies and DNA. The dogs were infested with ticks. Among the examined dogs only one (1.16%) had B. microti antibodies, associated to B. canis, A. phagocytophilum and R. conorii infections. In the PCR assay, the dog was also found positive for B. microti and R. conorii DNA, and negative for other microorganisms. The infected dog showed a non-specific flu-like syndrome, with depression, disorexia, hyperthermia (39.6°C), light dehydration, moderate lymphadenopathy and heavy tick infestation (>20). No significant changes were present in the cell blood count. To the best of our knowledge, this is the first report of serological and molecular identification of B. microti piroplasm in a dog from Southern Italy. Because specific antibodies and DNA were detected in an untraveled dog, babesiosis is probably due to an autochthonous tick infection. However, this study indicates that Babesia microti is not widely distributed in dog populations in Sicily, as demonstrated by low prevalence of infection. So, in Sicily the dog would not appear to represent a reservoir of infection, but rather an accidental host

Zoonotic infectious diseases in transplanted immunocompromised patients

Background. Immunocompromised patients, like transplant recipients, are a particularly vulnerable group being at higher risk of developing several infectious diseases. Among them, zoonotic diseases, such as visceral leishmaniasis, bartonellosis, Q fever and leptospirosis are a growing concern in immunosuppressed patients as they are more susceptible to develop severe symptoms of the diseases. Objectives. The study aimed at the detection of Leishmania infantum, Bartonella spp., Leptospira spp. and Coxiella burnetii DNA in immunocompromised hosts through molecular methods

Investigation of disease hazards in cattle in South of Italy (Sicily)

Istituto Zooprofilattico Sperimentale, Palermo, Italy. Objective: Infectious diseases represent a serious limitation of bovine production. The etiology of these diseases is diverse and comprises a variety of viral, bacterial, protozoan and chlamydial agents, some of which are zoonotic [1]. Infectious- parasitic agents associated with reproductive disorders in ruminants include Neospora caninum, Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii; they cause the greatest economic losses for the livestock industry. This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens including the zoonotic agents C. burnetii, T gondii, N. caninum, Clamydia spp. and Theileria annulata in cattle in Sicily region and to determine the molecular status for T. gondii in order to determine the serological and molecular status of bovine in the Sicily region

Messa a punto di una metodologia innovativa per l'identificazione molecolare delle principali specie di Culicoides - IZS SI 16/16 RC

Convegno Nazionale "I RISULTATI DELL’ATTIVITÁ DI RICERCA DELL’IZS SICILIA

Identification of Rickettsia species in ticks from ruminants in Lebanon

Rickettsia are tick-borne emerging pathogens recognized as important agents of human tick-borne diseases worldwide. Aim of this study was the identification and characterization of Spotted Fever Group (SFG) rickettsiae in ticks from ruminants in Lebanon. The study concerned 88 ticks collected in June 2014 from 30 Lebanese farms of ruminants and identified according morphological keys. Total DNA was extracted and used to identify and characterize Rickettsia spp. DNA through PCR and sequencing of fragments of 17 kDa protein, ompA, ompB, gltA, atpA, dnaK, dnaA, recA and 16S rRNA. Five different tick species were found: Hyalomma anatolicum (n=6), Rhipicephalus annulatus (n=56), Rhipicephalus bursa (n=1), Rhipicephalus sanguineus (n=6) and Rhipicephalus turanicus (n=19). A prevalence of 68.2% was found at the first screening performed amplyfing Rickettsia spp. 16S rRNA. Among these Rickettsia positive samples, 17 were identified at the species level and 43 as SFG rickettsiae based on the multigene genotyping strategy. ‘Candidatus Rickettsia barbariae’, an emerging member of the rickettsial SFG, was identified in 9 samples. R. massiliae, R. aeschlimannii and R. africae were identified in four, three and one tick, respectively, confirming the presence of SFG Rickettsia spp. involved in human diseases. SFG rickettsiae with public health relevance were found in ticks collected in Lebanon, where the widespread distribution of tick vectors and possible livestock animal hosts in contact with humans may favor transmission to humans. Funded by IZSSI 02/13, IZSSI 10/14, COMPARE, Grant 643476 and by UCLM Own Research Program. Thanks to Pippo Bono for technical support

Multispacer sequence typing of Coxiella burnetii from milk and hard tick samples from ruminant farms in Lebanon

his study was carried out to detect and characterize Coxiella burnetii in ruminant milk samples and in different tick species from seropositive farms in four Lebanese regions. Milk and tick samples were screened for C. burnetii presence by quantitative real-time PCR (qPCR) targeting IS1111 region followed by multispacer sequence typing (MST). The overall positive percentages of 9.6% (27/282) and 95.45% (84/88) for C. burnetii were recorded in ruminant milk and tick samples, respectively. In detail, the C. burnetii DNA was recorded in 52/54 (96.3%) of Rhipicephalus annulatus, 20/21 (95.24%) of Rhipicephalus turanicus, 6/6 (100%) of Hyalomma anatolicum, 5/6 (83.3%) of Rhipicephalus sanguineus and 1/1 of Rhipicephalus bursa. After genotyping of some IS1111-positive samples (17/111), different MST genotypes were identified. Out of 15 positive ticks, 10 were infected with MST2 genotype, 4 were infected with MST7 genotype and 1 was infected with MST57. Moreover, genotypes MST20 and MST58 were found in one cow and one goat milk samples, respectively. The present study confirmed the high genetic diversity of C. burnetii in Lebanon

Low NETosis Induced in Anaplasma phagocytophilumInfected Cells

Anaplasma phagocytophilum are obligatory intracellular bacteria that preferentially replicate inside leukocytes by utilizing biological compounds and processes of these primary host defensive cells. In this study, bioinformatics analysis was conducted to further characterize A. phagocytophilum–host interactions using the neutrophil-like model of human Caucasian promyelocytic leukemia HL60 cells. We detected a hierarchy of molecules involved in A. phagocytophilum-HL60 interactions with overrepresentation in infected human cells of proteins involved in the reactive oxygen species (ROS) pathway and cell surface monocyte markers. As A. phagocytophilum phagocytosis by neutrophils is inhibited, the results suggested a possible explanation for our bioinformatics data: radical oxygen compounds could induce the killing of bacteria activating NETosis, a unique form of defense mechanism resulting in cell death that is characterized by the release of decondensed chromatin and granular contents to the extracellular space, forming neutrophil extracellular traps (NETs) to eliminate invading microorganisms. Thus, we confirmed the existence of a low NETosis induced in A. phagocytophilum-infected cells by immunofluorescence (IF) experiments. These results provide new insights into the complex mechanisms that govern immune response during A. phagocytophilum host interactions.Anaplasma phagocytophilum son bacterias intracelulares obligatorias que se replican preferentemente dentro de los leucocitos mediante la utilización de compuestos biológicos y procesos de estas células defensivas primarias del huésped. En este estudio, se realizó un análisis bioinformático para caracterizar aún más las interacciones entre A. phagocytophilum y el huésped utilizando el modelo similar a los neutrófilos de células HL60 de leucemia promielocítica caucásica humana. Detectamos una jerarquía de moléculas involucradas en las interacciones A. phagocytophilum -HL60 con una sobrerrepresentación en células humanas infectadas de proteínas involucradas en la vía de especies reactivas de oxígeno (ROS) y marcadores de monocitos de superficie celular. Como A. phagocytophilumse inhibe la fagocitosis de los neutrófilos, los resultados sugirieron una posible explicación para nuestros datos bioinformáticos: los compuestos radicales de oxígeno podrían inducir la muerte de bacterias activando NETosis, una forma única de mecanismo de defensa que resulta en la muerte celular que se caracteriza por la liberación de cromatina descondensada y granular. contenido al espacio extracelular, formando trampas extracelulares de neutrófilos (NET) para eliminar los microorganismos invasores. Por lo tanto, confirmamos la existencia de una NETosis baja inducida en células infectadas por A. phagocytophilum mediante experimentos de inmunofluorescencia (IF). Estos resultados proporcionan nuevos conocimientos sobre los complejos mecanismos que gobiernan la respuesta inmunitaria durante las interacciones del huésped con A. phagocytophilum

Biotic and abiotic factors shape the microbiota of wild-caught populations of the arbovirus vector Culicoides imicola

Biting midges of the genus Culicoides are known vectors of arboviruses affecting human and animal health. However, little is known about Culicoides imicola microbiota and its influence on this insect’s biology. In this study, the impact of biotic and abiotic factors on C. imicola microbiota was characterized using shotgun-metagenomic sequencing of whole-body DNA samples. Wild-caught C. imicola adult nulliparous females were sampled in two locations from Sicily, Italy. The climatic variables of temperature and soil moisture from both localities were recorded together with potential host bloodmeal sources. Shared core microbiome among C. imicola populations included Pseudomonas, Escherichia, Halomonas, Candidatus Zinderia, Propionibacterium, and Schizosaccharomyces. Specific and unique taxa were also found in C. imicola from each location, highlighting similarities and differences in microbiome composition between the two populations. DNA and protein identification showed differences in host preferences between the two populations, with Homo sapiens and Canis lupus familiaris L. being the preferred bloodmeal source in both locations. A principal component analysis showed that the combined effect of host preferences (H. sapiens) and local soil moisture factors shape the microbiome composition of wild-caught populations of C. imicola. These results contribute to characterizing the role of the microbiome in insect adaptation and its utility in predicting geographic expansion of Culicoides species with potential implications for the control of vector-borne diseases.This work was financially supported by the H2020 COllaborative Management Platform for detection and Analyses of (Re-) emerging and foodborne outbreaks in Europe (COMPARE) grant 643476, and the Italian Ministry of Health grants RC IZS SI 01/13 and RC IZS SI 03/15.Peer reviewe

Infection and exposure to vector-borne pathogens in rural dogs and their ticks, Uganda

BACKGROUND: In rural parts of Africa, dogs live in close association with humans and livestock, roam freely, and usually do not receive prophylactic measures. Thus, they are a source of infectious disease for humans and for wildlife such as protected carnivores. In 2011, an epidemiological study was carried out around three conservation areas in Uganda to detect the presence and determine the prevalence of vector-borne pathogens in rural dogs and associated ticks to evaluate the risk that these pathogens pose to humans and wildlife. METHODS: Serum samples (n = 105), blood smears (n = 43) and blood preserved on FTA cards (n = 38) and ticks (58 monospecific pools of Haemaphysalis leachi and Rhipicephalus praetextatus including 312 ticks from 52 dogs) were collected from dogs. Dog sera were tested by indirect immunofluorescence to detect the presence of antibodies against Rickettsia conorii and Ehrlichia canis. Antibodies against R. conorii were also examined by indirect enzyme immunoassay. Real time PCR for the detection of Rickettsia spp., Anaplasmataceae, Bartonella spp. and Babesia spp. was performed in DNA extracted from FTA cards and ticks. RESULTS: 99 % of the dogs were seropositive to Rickettsia spp. and 29.5 % to Ehrlichia spp. Molecular analyses revealed that 7.8 % of the blood samples were infected with Babesia rossi, and all were negative for Rickettsia spp. and Ehrlichia spp. Ticks were infected with Rickettsia sp. (18.9 %), including R. conorii and R. massiliae; Ehrlichia sp. (18.9 %), including E. chaffeensis and Anaplasma platys; and B. rossi (1.7 %). Bartonella spp. was not detected in any of the blood or tick samples. CONCLUSIONS: This study confirms the presence of previously undetected vector-borne pathogens of humans and animals in East Africa. We recommend that dog owners in rural Uganda be advised to protect their animals against ectoparasites to prevent the transmission of pathogens to humans and wildlife