Association between canine leishmaniosis and Ehrlichia canis co-infection: a prospective case-control study

Abstract Background In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. Methods We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. Results From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or “Candidatus Mycoplasma haematoparvum” DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5–106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to be co-infected with M. haemocanis. Conclusions Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR

Serological survey of Chlamydia abortus in Greek dairy sheep flocks

Chlamydiosis due to Chlamydia abortus is one of the most common causes of abortion in small ruminant flocks worldwide. Although the causative agent is zoonotic, chlamydiosis is not a reportable disease. There is lack of recent data concerning sheep chlamydiosis in Greece.In the current study, a serological investigation for Chlamydia abortus was conducted. Blood samples from 26 randomly selected sheep flocks not vaccinated against chlamydiosis were collected. From each flock, 15 to 20 blood samples were taken from adult female sheep. In total 464 blood samples were examined. One hundred and six samples were positive (22.8%), while 24 samples (5.24%) were doubtful.  Moreover, at farm level, in 18 out of the total number of 26 farms, at least one positive animal was detected (69.2%).Chlamydiosis is considered a highly likely cause of sheep abortion in Greece. Therefore, vaccination of the sheep flocks is strongly recommended for the prevention and control of the disease.

Electrochemical Antigenic Sensor for the Diagnosis of Chronic Q Fever

In this work, we report the development of an impedimetric biosensor for the direct, quick, and easy diagnosis of chronic Q fever. The biosensor is based on highly sensitive antigens that can selectively recognize antibodies against Coxiella burnetii. The biosensor is based on the immobilization of antigens onto a gold electrode using the EDC/NHS immobilization methodology. The detection is performed by impedance spectroscopy that monitors specific frequencies which provide the maximum sensitivity for the biosensor. Q fever antibodies that are present in the sera of patients interact selectively with the biosensor antigens, thereby altering the impedance of the biosensor surfaceand generating a large impedance change within a few seconds. The biosensor allows for the specific serological detection of chronic Q fever, while the developed system can also be modified for the detection of other biomarkers, such as the ones against acute Q fever

Molecular detection of tick-borne bacteria and protozoa in cervids and wild boars from Portugal

Background: Wildlife can act as reservoir of different tick-borne pathogens, such as bacteria, parasites and viruses. The aim of the present study was to assess the presence of tick-borne bacteria and protozoa with veterinary and zoonotic importance in cervids and wild boars from the Centre and South of Portugal.Methods: One hundred and forty one blood samples from free-ranging ungulates including 73 red deer (Cervus elaphus), 65 wild boars (Sus scrofa) and three fallow deer (Dama dama) were tested for the presence of Anaplasma marginale/A. ovis, A. phagocytophilum, Anaplasma/Ehrlichia spp., Babesia/Theileria spp., Borrelia burgdorferi (sensu lato) (s.l.), and Rickettsia spp. DNA by PCR.Results: Anaplasma spp. DNA was detected in 33 (43.4 %) cervids (31 red deer and two fallow deer) and in two (3.1 %) wild boars while Theileria spp. were found in 34 (44.7 %) cervids (32 red deer and two fallow deer) and in three (4.6 %) wild boar blood samples. Sequence analysis of msp4 sequences identified A. marginale, A. ovis, while the analysis of rDNA sequence data disclosed the presence of A. platys and A. phagocytophilum and T. capreoli and Theileria sp. OT3. Anaplasma spp./Theileria spp. mixed infections were found in 17 cervids (22.4 %) and in two wild boars (3.1 %). All samples were negative for Babesia sp., B. burgdorferi (s.l.), Ehrlichia sp. or Rickettsia sp.Conclusions: This is the first detection of Anaplasma marginale, A. ovis, A. phagocytophilum, A. platys, Theileria capreoli and Theileria sp. OT3 in cervids and wild boars from Portugal. Further studies concerning the potential pathogenicity of the different species of Anaplasma and Theileria infecting wild ungulates, the identification of their vector range, and their putative infectivity to domestic livestock and humans should be undertaken

Bartonella vinsonii sub. arupensis infection in animals of veterinary importance, ticks and biopsy samples

Testing for vector-borne pathogens in livestock is largely reliant upon blood and tissue. The role of biopsy samples remains poorly explored for detecting tick-borne bacteria in animals.In a 2-year survey, animals of veterinary importance from farms throughout the Northern part of Greece were routinely checked for the presence of biopsy samples. Where detected, either a portion or biopsy was collected together with whole blood samples and any ticks at the site of the biopsy sample. Molecular testing was carried out by real-time PCR targeting the ITS gene of Bartonella species.A total 68 samples [28 blood samples, 28 biopsy samples and 12 ticks (9 Rhipicephalus bursa and 3 R. turanicus)] were collected from goats (64 samples) and bovine (4 samples).Eight (11.8%) of the 68 samples were positive for Bartonella species. Of the biopsy sample and whole blood samples, four (14.3%) of each type were positive for Bartonella species. None of the ticks was tested positive for Bartonella species. All pairs of positive biopsy samples/whole blood samples originated from the same animals. Positive samples were identified as B. vinsonii sub. arupensis.Although many more samples from a much wider spectrum of animal species is required before concluding upon the merit of biopsy samples on the study of tick-borne diseases, the significance of our finding warrants further study, both for clinical consequences in small ruminants and for those humans farming infected animals