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

PurposeDermacentor 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 methodsTicks 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.ResultsTick-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.ConclusionsDermacentor 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.     

Co-infections with Borrelia species,Anaplasma phagocytophilum and Babesia spp. in patients with tick-borne encephalitis

The purpose of this study was evaluation of the prevalence of co-infection with Borrelia species, A. phagocytophilum and Babesia spp. in patients with tick-borne encephalitis (TBE). At total of 110 patients with TBE were included in the study. Serological tests for tick-borne encephalitis virus (TBEV), PCR for Borrelia species, Anaplasma phagocytophilum and Babesia spp., blood smears for A. phagocytophilum and Babesia spp. and BLAST analysis for Babesia spp. were performed. Results showed a significant majority of patients co-infected with Borrelia species (30/110; 27 %), much less with A. phagocytophilum (12/110; 10.9 %) and with Babesia spp. (1/110; 0.9 %). The BLAST analysis of the 18S rDNA sequence obtained with the Babesia spp. specific primers indicated that the patient was infected with Babesia microti. Triple co-infections (TBEV-Borrelia species- A. phagocytophilum) were observed in three (3/110; 2.7 %) patients. Conclusions were such that differential diagnosis in patients after the tick bite, presenting with acute symptoms, should include not only TBE and Lyme disease, but also other diseases transmitted by ticks. In patients with low parasitemia in suspicion of Babesia spp. infection PCR seems to be a more sensitive method than blood smear. Co-infection with various tick-borne pathogens must be always considered, especially in endemic regions.     

Identification of<Emphasis Type="Italic"> Borrelia burgdorferi</Emphasis> sensu lato,<Emphasis Type="Italic"> Anaplasma</Emphasis> and<Emphasis Type="Italic"> Ehrlichia</Emphasis> Species,and Spotted Fever Group Rickettsiae in Ticks from Southeastern Europe

Prevalence data for tick-borne pathogens are used to assess the risk for human health. In this study the presence and identity of Borrelia burgdorferi sensu lato, Ehrlichia, Anaplasma, and Rickettsia species in Bulgarian Ixodes ricinus ticks and in non-Ixodes ticks from Turkey and Albania was determined by polymerase chain reaction (PCR) and reverse line blot hybridization. In the adult Bulgarian ticks, the prevalence of Borrelia burgdorferi sensu lato infection was approximately 40%, while Borrelia afzelii was the predominant species, representing more than half of all Borrelia-positive ticks. Ehrlichia and Anaplasma species were detected in 35% of the adult Ixodes ricinus ticks and in 10% of the nymphs. Sequence analysis of PCR products reacting with the Anaplasma phagocytophila probe revealed a 16S rRNA gene identical to that of the Anaplasma phagocytophila prototype strain. Ehrlichia and Anaplasma species were found in approximately 7% of the non-Ixodes ticks. Sequence analysis of some of these samples revealed the presence of Anaplasma ovis, Ehrlichia canis, and a species closely resembling Ehrlichia chaffeensis. About half of all adult ticks examined and approximately 20% of all nymphs were infected with Rickettsia species. In Ixodes ricinus ticks, Rickettsia helvetica and a Rickettsia species designated as IRS3 were found in high prevalence. Rickettsia conorii was found in virtually all non-Ixodes tick species from Albania and Turkey. The results of this study show that many tick-borne diseases are most probably endemic in the Balkan area. Furthermore, the results suggest that there is a considerable chance for simultaneous transmission of tick-borne pathogens to human beings.     

Clinical,serological, and molecular evidence of ehrlichiosis and anaplasmosis in dogs in Tunisia

A seroepidemiological survey was conducted in five bioclimatic areas of Tunisia to determine the prevalence of antibodies to Ehrlichia canis and Anaplasma phagocytophilum antigens, surrogate markers of the agents of canine monocytic ehrlichiosis (CME) and canine granulocytic ehrlichiosis, respectively. Among 286 collected sera, 54.2% and 25.2% were seropositive for E. canis and A. phagocytophilum, respectively, by the indirect immunofluorescence antibody (IFA) test. Clinical and hematological tests were done only for 58 sick dogs from Tunis area. A reverse line blot (RLB) hybridization was then used to identify isolated Ehrlichia and Anaplasma species infecting dogs (n = 228). Among them, only two dogs were infected by A. phagocytophilum; ten sample dogs were demonstrated infected by E. canis and ten infected by Ehrlichia sp., from which one dog showed a mixed infection with A. phagocytophilum and E. canis and one with A. phagocytophilum and Ehrlichia sp. RLB findings were confirmed by sequencing; BLAST search against GenBank revealed high similarity of the sequence of Ehrlichia sp. PCR/RLB amplicons with Anaplasma platys 16S rRNA partial sequence.     

Prevalence and diversity of Babesia,Hepatozoon, Ehrlichia,and Bartonella in wild and domestic carnivores from Zambia,Africa

A molecular survey was conducted for several hemoparasites of domestic dogs and three species of wild carnivores from two sites in Zambia. Three Babesia spp. were detected including Babesia felis and Babesia leo in lions (Panthera leo) and a Babesia sp. (similar to Babesia lengau) in spotted hyenas (Crocuta crocuta) and a single lion. All wild dogs (Lycaon pictus) and domestic dogs were negative for Babesia. High prevalences for Hepatozoon were noted in all three wild carnivores (38–61 %) and in domestic dogs (13 %). Significantly higher prevalences were noted in hyenas and wild dogs compared with domestic dogs and lions. All carnivores were PCR negative for Ehrlichia canis, Ehrlichia ewingii, and Bartonella spp. Overall, high prevalences and diversity of Babesia and Hepatozoon were noted in wild carnivores from Zambia. This study is the first molecular characterization of Babesia from any hyena species and is the first report of a Babesia sp. closely related to B. lengau, a parasite previously only reported from cheetahs (Acinonyx jubatus), in lions and hyenas. Although usually benign in wild carnivores, these hemoparasites can be pathogenic under certain circumstances. Importantly, data on vectors for these parasites are lacking, so studies are needed to identify vectors as well as determine transmission routes, infection dynamics, and host specificity of these hemoparasites in wildlife in Africa and also the risk of transmission between domestic animals and wildlife.     

Seroprevalence of vector-borne pathogens in dogs from Croatia

Canine vector-borne pathogens (VBPs) are a group of globally distributed and rapidly spreading microorganisms transmitted by arthropods. In the present survey, we investigated Anaplasma spp., Ehrlichia canis, Dirofilaria immitis and Borrelia burgdorferi sensu lato seroprevalence between three groups of dogs (asymptomatic, suspected and deceased) from continental and coastal regions of Croatia with the commercial point-of-care SNAP®4Dx®Plus for VBPs. The overall prevalence for tested pathogens in dogs was 6.1% with the highest prevalence detected for Anaplasma spp. (4.5%), while the remaining pathogens were found at a prevalence of less than 1% (E. canis 0.6%, B. burgdorferi s.l. 0.4%, D. immitis 0.6%). No statistically significant differences in VBP detection between dog cohorts could be found with the exception of D. immitis in the deceased group. Interestingly, no evidence of D. immitis could be found in the hearts of dogs in this group at necropsy, however. This study provides the first data on the seroprevalence of selected VBPs between dogs of different health statuses in Croatia. The results demonstrate that serological evidence of VBPs alone or in combination with co-infections were found just as frequently in asymptomatic dogs as those with suspected or confirmed evidence of VBP disease, raising questions about the pathogenic potential of these organisms in domesticated dogs.

     

Serological detection of Anaplasma phagocytophilum,Borrelia burgdorferi sensu lato and Ehrlichia canis antibodies and Dirofilaria immitis antigen in a countrywide survey in dogs in Poland

Canine vector-borne diseases (CVBDs) have increasingly become a focus of attention in the past few years. Nevertheless, in many parts of Europe information on their occurrence is still scarce. In a large study in Poland 3,094 serum samples taken from dogs throughout all 16 Polish provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen. A total of 12.31 % (381/3,094; 95 % confidence interval [CI]: 11.18–13.52 %) and 3.75 % (116/3,094; 95 % CI: 3.11–4.48 %) of the dogs were positive for A. phagocytophilum and B. burgdorferi s.l. antibodies, respectively. Furthermore, 0.26 % (8/3,094; 95 % CI: 0.11–0.51 %) were positive for E. canis antibodies and 0.16 % (5/3,094; 95 % CI: 0.05–0.38 %) for D. immitis antigen. The highest percentages of A. phagocytophilum-positive dogs were noted in Lesser Poland, Silesia and ?ód? Provinces. For B. burgdorferi s.l., the highest prevalence was recorded in ?ód? Province. Co-infections with A. phagocytophilum and B. burgdorferi s.l. were recorded in 1.71 % of all examined dogs (53/3,094; 95 % CI: 1.29–2.23 %). One dog even had a triple infection, testing positive for E. canis too. Both A. phagocytophilum and B. burgdorferi s.l. have previously been reported in Poland and were confirmed in the present study by positive samples from all 16 provinces. Concerning E. canis and D. immitis travel history or importation cannot be excluded as factors which may have determined the occurrence of these pathogens in the relevant animals. Practitioners in Poland should be aware of the above mentioned CVBDs and of prophylactic measures to protect dogs and their owners.     

PCR-detection and molecular-genetic analysis of tick-transmitted pathogens in patients of Novosibirsk region,Russia

Blood samples of patients collected after tick bite in Novosibirsk region, Russia, in spring–summer of 2003–2004 were examined for tick-borne encephalitis virus (TBEV), Borrelia, Anaplasma, Babesia, and Rickettsia by (RT)-PCR. TBEV of the Far-Eastern subtype, Bo. garinii, Bo. afzelii, and Rickettsia sp. closely related to R. sibirica but not Anaplasma sp. or Babesia sp. were found.     

Sequential Evaluation of Dogs Naturally Infected with Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, Ehrlichia ewingii, or Bartonella vinsonii

Historically, disease manifestations in dogs seroreactive to Ehrlichia canis antigens by indirect immunofluorescent antibody testing have been attributed to infection with either E. canis or Ehrlichia ewingii. A 1996 study by Dawson and colleagues provided PCR evidence that healthy dogs from southeastern Virginia could be naturally infected with Ehrlichia chaffeensis. This observation stimulated us to determine which Ehrlichia spp. infected sick dogs that were referred to our hospital from the same region. Based upon PCR amplification with species-specific primers, sick dogs seroreactive to E. canis antigens were determined to be infected with four Ehrlichia species: E. canis, E. chaffeensis, E. equi, and E. ewingii. Coinfection with three Ehrlichia species (E. canis, E. ewingii, and E. equi) was documented for one dog. An additional canine pathogen presumed to be tick transmitted, Bartonella vinsonii subsp. berkhoffii, was identified in 7 of 12 dogs. Importantly, our results indicate that in naturally infected dogs, E. chaffeensis can cause severe disease manifestations that are clinically and serologically indistinguishable from disease manifestations of E. canis or E. ewingii. In addition, our findings support the efficacy of doxycycline for treatment of E. canis, E. equi, and E. ewingii infections but indicate that, based upon the persistence of E. chaffeensis DNA for 1 year following treatment, E. chaffeensis infection in dogs may be more refractory to doxycycline treatment. Undetected coinfection with Bartonella may also complicate the evaluation of treatment efficacy while resulting in disease manifestations that mimic ehrlichiosis.     

Prevalence of Rickettsiales (Anaplasma phagocytophilum and Rickettsia spp.) in hard ticks (Ixodes ricinus) in the city of Hamburg,Germany

To narrow the gap of missing knowledge on Rickettsia spp. and Anaplasma phagocytophilum infections in ticks in northwestern Germany and, at the same time, to provide first prevalence data on these pathogens in the city of Hamburg, a total of 1,400 questing Ixodes ricinus ticks were collected at ten different public green areas from April until October 2011. Ticks were examined using probe-based quantitative real-time PCR. A percentage of 3.6 % (51/1,400) ticks were tested positive for A. phagocytophilum infections divided into 2.1 % (3/141) adults [1.7 % (1/60) females and 2.5 % (2/81) males] and 3.8 % (48/1,259) nymphs. The percentage of infected ticks per sampling site varied statistically significantly from 0.7 % (1/140) to 12.1 % (17/140), whereas between sampling months, no statistically significant differences were observed (2.0–6.5 %, 4–13/140). The overall Rickettsia spp. infection rate was 52.5 % (735/1,400). In adult ticks, Rickettsia spp. infection rate was 56 % (79/141) divided into 61.7 % (37/60) infected females and 51.9 % (42/81) infected males. Nymphs showed an infection rate of 52.1 % (656/1,259). In contrast to A. phagocytophilum infections, no statistically significant differences in Rickettsia spp. infection rates among sampling sites (44.3–63.6 %, 62–89/140) were observed, whereas seasonal variations were obvious: the percentage of Rickettsia-positive ticks was significantly lower in April (36.5 %, 73/200) and May (29.5 %, 59/200) compared to the summer and fall months (55.0–64.5 %, 110–129/200). Rickettsia species differentiation via real-time pyrosequencing revealed Rickettsia helvetica as the only occurring species. Co-infections with both Rickettsia spp. and A. phagocytophilum were detected in 2.0 % (28/1,400) of the ticks. The present study revealed that in the city of Hamburg, the tick infection rate with A. phagocytophilum is comparable with other German data, whereas the Rickettsia spp. infection rate of 52.5 % is by far the highest prevalence detected in Germany so far. As the city of Hamburg has 1.8 million inhabitants and attracts millions of tourists every year, the potential health risk should not be underestimated.     

Seroreactivity for spotted fever rickettsiae and co-infections with other tick-borne agents among habitants in central and southern Sweden

Patients seeking medical care with erythema migrans or flu-like symptoms after suspected or observed tick bite in the southeast of Sweden and previously investigated for Borrelia spp. and/or Anaplasma sp. were retrospectively examined for serological evidence of rickettsial infection (Study 1). Twenty of 206 patients had IgG and/or IgM antibodies to Rickettsia spp. equal to or higher than the cut-off titre of 1:64. Seven of these 20 patients showed seroconversion indicative of recent or current infection and 13 patients had titres compatible with past infection, of which five patients were judged as probable infection. Of 19 patients with medical records, 11 were positive for Borrelia spp. as well, and for Anaplasma sp., one was judged as positive. Five of the 19 patients had antibodies against all three pathogens. Erythema migrans or rash was observed at all combinations of seroreactivity, with symptoms including fever, muscle pain, headache and respiratory problems. The results were compared by screening an additional 159 patients (Study 2) primarily sampled for the analysis of Borrelia spp. or Mycoplasma pneumoniae. Sixteen of these patients were seroreactive for Rickettsia spp., of which five were judged as recent or current infection. Symptoms of arthritis, fever, cough and rash were predominant. In 80 blood donors without clinical symptoms, approximately 1 % were seroreactive for Rickettsia spp., interpreted as past infection. The study shows that both single and co-infections do occur, which illustrate the complexity in the clinical picture and a need for further studies to fully understand how these patients should best be treated.     

Prevalence and diversity of Hepatozoon canis in naturally infected dogs in Japanese islands and peninsulas

Canine hepatozoonosis is a worldwide protozoal disease caused by Hepatozoon canis and Hepatozoon americanum and is transmitted by ixodid ticks, Rhipicephalus and Amblyomma spp., respectively. H. canis infection is widespread in Africa, Europe, South America, and Asia, including Japan. The objective of this study was to study the distribution pattern and diversity of H. canis in naturally infected dogs in nine Japanese islands and peninsulas. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011 and the ages and sexes were identified. Direct microscopy using Giemsa-stained blood smears revealed H. canis gametocytes in the peripheral blood of 45 (23.6 %) dogs. Polymerase chain reaction (PCR) was performed on EDTA-anticoagulated blood, initially with the common primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, and then with the specific primer set (HepF and HepR) amplifying about 660 bp fragments of the same gene. Based on PCR, 84 (42.9 %) dogs were positive using the common primer and 81 (41.3 %) were positive using the specific primer. The current investigation indicated that all screened areas, except for Sado Island and Atsumi Peninsula, were infected. Yaku Island had the highest infection rate (84.6 % in males and 100.0 % in females), while Ishigaki Island showed the lowest infection rates (8.3 % in males and 17.7 % in females). Both sexes were infected with no significant difference. However, diversity of infection among the surveyed islands and peninsulas was significantly different (P?<?0.05). Although H. canis has previously been reported in dogs in Japan, the higher infection rate described in the current study and the diversity of infection in a wide range of islands strongly encourage prospective studies dealing with the prevention and treatment of the infection in dogs, as well as control of ticks.     

Skin and soft-tissue infections caused by Aeromonas species

This study investigated the clinical characteristics of patients with skin and soft-tissue infections (SSTIs) due to Aeromonas species. Patients with SSTIs caused by Aeromonas species during the period from January 2009 to December 2011 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed. A total of 129 patients with SSTIs due to Aeromonas species were identified. A. hydrophila (n?=?77, 59.7 %) was the most common pathogen, followed by A. veronii biovar sobria (n?=?22, 17.1 %), A. veronii biovar veronii (n?=?20, 15.5 %), A. caviae (n?=?9, 7.0 %), and A. schubertii (n?=?1, 0.8 %). The most common isolates obtained from patients with polymicrobial infections were Klebsiella species (n?=?33), followed by Enterococcus spp. (n?=?24), Enterobacter spp. (n?=?21), Escherichia coli (n?=?17), Staphylococcus spp. (n?=?17), Streptococcus spp. (n?=?17), and Acinetobacter spp. (n?=?15). Liver cirrhosis and concomitant bacteremia were more common among patients with monomicrobial Aeromonas SSTIs than among patients with polymicrobial SSTIs. Nine (7 %) patients required limb amputations. The in-hospital mortality rate was 1.6 %. In conclusion, Aeromonas species should be considered as important causative pathogens of SSTIs, and most infections are polymicrobial. In addition, the clinical presentation differs markedly between patients with monomicrobial and those with polymicrobial Aeromonas SSTIs.     

Molecular investigations of Hepatozoon species in dogs and developmental stages of Rhipicephalus sanguineus

The occurrence and distribution of Hepatozoon species in stray dogs, and the developmental stages of Rhipicephalus sanguineus detached from the same dogs in Diyarbak?r Province, Turkey is reported. A total of 328 ticks, including 133 adults (55 males and 75 females consist of 63 partially engorged and 15 fully engorged) and 195 nymphs (91 partially engorged and 104 fully engorged) were detached from the dogs. Fully engorged nymphs and females were incubated at 27 °C and relative humidity of 85 % to molt to adult stage and recover eggs. The ticks were pooled according to sex and developmental stage. No Hepatozoon gamonts were found, whereas, by PCR, 15.87 % (10/63) of the dogs were infected with Hepatozoon canis. Of the 68 tick pools tested, 14 (20.58 %) pools were infected with Hepatozoon spp., an overall maximum likelihood estimation of prevalence of 4.9 % (95 % confidence intervals (CI)?=?2.85–7.93 %) per 100 ticks. Maximum likelihood estimation of the infection rate varied by tick sex and developmental categories, ranging from 1.75 % (95 % CI?=?0.11–8.11 %) in fed males to 6.81 % (95 % CI?=?2.07–17.46 %) in unfed females. One amplicon from a fed adult female was 99 % identical to the sequence for Hepatozoon felis. The remaining sequences isolated from both dogs and ticks shared 99–100 % similarity with the corresponding H. canis isolates. This is the first detection of H. canis and H. felis in the tick R. sanguineus in Turkey.     

Brucella canis-induced anterior uveitis in companion dogs in Ahvaz,Iran

Canine brucellosis, caused by Brucella canis, is an important cause of abortions, stillbirths, uveitis, epididymitis, orchitis, and sperm abnormalities in dogs. Anterior uveitis is one of the most common ocular diseases in dogs. The aim of this study was to describe the clinical and diagnostic features of anterior uveitis in dogs affected with B. canis in the Ahvaz district, southwestern Iran. A total of 243 blood samples were obtained from urban dogs between 2006 and 2010 and examined by immuno-chromatography assay (Anigen Rapid C. Brucella Ab Test Kit). Prevalence to B. canis antibodies was 6.58 % (16 out of 243) in the studied dogs. B. canis-induced anterior uveitis was seen in two German Shepherd dogs (12.5 %; 2 out of 16), the first was a 3.5-year-old male and the second was a 2.5-year-old female. Clinical ophthalmologic abnormalities included moderate anterior uveitis, mild conjunctival hyperemia, and iris hyperpigmentation. Following routine treatment (a median of 9 weeks from the onset of therapy), ocular inflammation resolved and serological tests were negative. The present study shows that ocular inflammation due to B. canis is present among the urban dog population in the Ahvaz district, Iran. B. canis infection should be included in the differential diagnosis for dogs with intraocular inflammation, regardless of previous history.     

Canine packed red blood cell transfusions in Spain

Transfusion medicine is a relatively new and rapidly growing area of research in veterinary medicine. Packed red blood cell transfusion (PRBC) is indicated for treatment of symptomatic anemia resulting from hemorrhage, hemolysis, or ineffective erythropoiesis. The objective of this retrospective study was to identify clinical manifestations and underlying diseases of dogs that received PRBC and determine possible transfusion complications and outcome. Donors were blood typed and previously tested for infectious diseases potentially transmitted by transfusion (Ehrlichia canis, Borrelia burdogferi, Dirofilaria immitis, Anaplasma phagocytophila, Anaplasma platys, Babesia spp., Bartonella spp., and Rickettsia spp, and Leishmania infantum). Recipients were also blood typed and cross-matching was routinely performed before any transfusion. Packed cell volume (PCV) was performed before and after transfusion. Every PRBC transfusion was delivered by a bedside leukoreduction filter. Sixty-five PRBC transfusions were administered to 56 dogs. Twenty-two dogs resulted DEA 1.1 positive and 34 DEA 1.1 negative. Reasons for transfusion included anemia secondary to hemorrhage (n = 48; 74%), hemolysis (n = 8; 12%), and ineffective erythropoiesis (n = 9; 14%). Median PCV before transfusion was 14.7% (range: 7–36%) and the mean post-transfusion was 21% (range: 9–39%). Mean increase in PCV was 6.5%. Thirty-one (70%) dogs were discharged and 17 (30%) dogs died or were euthanized. Transient hyperthermia was the only adverse reaction found. PRBC transfusion for symptomatic treatment of anemia is a safe and useful procedure, if the transfusion is closely supervised throughout its duration.     

Detection of Rickettsia in Rhipicephalus sanguineus Ticks and Ctenocephalides felis Fleas from Southeastern Tunisia by Reverse Line Blot Assay

Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia.     

Human babesiosis in Germany: Just overlooked or truly new?

Tick-borne infections are among the more important vector-borne infections in the northern hemisphere. However, many facts pertaining to the epidemiology and pathogenesis of such diseases in Europe remain unclear. Human babesiosis in particular may have previously been overlooked in many parts of the world due to a lack of medical awareness and microbiological detection methods. Recently, the first two cases of human babesiosis were reported in Germany, occurring in vicinities where the presence of Babesia spp. in enzootic cycles was obvious for decades but where the risk of acquiring Babesia spp. either from ticks or from human blood products was not known before. It is important to note, though, that as with other tick-borne diseases, Babesia infections may arise in geographic areas where they have not been recorded in the past. Better molecular detection methods and strain typing of parasites are necessary to investigate the epidemiological distribution of zoonotic Babesia spp. in Europe and to clarify whether their virulence or transmissibility is strain-dependent. Therefore, further seroepidemiological and molecular epidemiological studies are urgently needed to learn more about the distribution and medical relevance of these pathogens in various parts of Europe in general and in Germany in particular.     

Canine babesiosis in Romania due to <Emphasis Type="Italic">Babesia canis</Emphasis> and <Emphasis Type="Italic">Babesia vogeli</Emphasis>: a molecular approach

Canine babesiosis is a tick-borne disease caused by the protozoa Babesia spp. that affects dogs worldwide. In Romania, canine babesiosis has become quite frequent in the last few years, with a wide variety of clinical signs, ranging from mild, nonspecific illness to peracute collapse, and even death. Traditionally, a Babesia infection in dogs is diagnosed based on the morphologic appearance of the intraerythrocytic piroplasms observed in peripheral blood smears. To date, no data on genetic characterization of Babesia species in dogs has been documented for Romania. Therefore, a molecular survey on natural Babesia infections of dogs in Romania using polymerase chain reaction and genetic sequence analysis of a fragment of the ssRNA gene was performed. A total number of 16 blood samples were tested for the presence of Babesia DNA. Blood samples were collected from 11 dogs with symptoms of babesiosis and microscopically proven positive for Babesia and from a group of five asymptomatic dogs, not tested microscopically for Babesia, which were included in the study for comparative analysis. The piroplasm-specific PCR amplifying the partial 18S rRNA gene confirmed Babesia spp. infection in all 11 samples from dogs with clinical babesiosis, and in one of the clinically normal dogs. Sequence analysis revealed the presence of Babesia canis in all clinically affected dogs and Babesia vogeli in one clinically normal dog. This is the first molecular evidence of B. canis and B. vogeli in dogs from Romania. The results of the study provide basic information toward a better understanding of the epidemiology of canine babesiosis in Romania and will help to promote an effective control program.     

Prevalence of Anaplasma and Ehrlichia species in Ixodes persulcatus ticks and small mammals from different regions of the Asian part of Russia

A total 834 Ixodes persulcatus ticks and 641 small mammals from the areas of northern and southern Ural, southeastern and southwestern Siberia, and the Far East was examined on the presence of Anaplasma spp. and Ehrlichia spp. by nested PCR based on 16S rRNA gene with subsequent sequencing. A. phagocytophilum and E. muris DNAs were found in samples from ticks and small mammals in all studied areas. A. phagocytophilum DNA was revealed in 0.3–5.1% of ticks and 3.2–16.6% of small mammals from different regions, whereas E. muris DNA was found in 4.9–15.3% of ticks and 4.1–12.6% of small mammals. One I. persulcatus tick and one east-European field vole from the Siberian region were shown to contain ‘Candidatus Neoehrlichia mikurensis’ DNA. A novel Ehrlichia species, closely related to Ehrlichia sp. EHf669, Ehrlichia sp. EH727, Ehrlichia sp. Tibet and ‘Candidatus Ehrlichia shimanensis’, was found using molecular techniques in tissue samples of 14.4% of small rodents from the Far East.