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Canine babesiosis has been increasingly diagnosed in various regions of Germany such as north-eastern Germany in recent years. A dog with several relapses of Babesia canis infection after treatment with imidocarb is described. A 9-year-old male Magyar Viszla with B. canis infection was referred after two treatments with imidocarb (dosage 2.1 mg/kg SC) because of lethargy, fever and pancytopenia (additional treatments with prednisolone and doxycycline). Merozoites were detected in the blood smear and imidocarb treatment was repeated. Clinical signs, pancytopenia and a positive B. canis PCR occurred after the 3rd (6 mg/kg SC), 4th (7.7 mg/kg SC) and 5th (7.5 mg/kg SC and doxycycline for 4 weeks in addition) imidocarb injection and thorough tick prevention with isoxazoline and permethrin products. 12 days after the 5th injection, the PCR was negative for the first time. The dog was again presented with fever 35 days after the 5th injection. The B. canis PCR was positive and laboratory examination revealed pancytopenia. Treatment with atovaquone/azithromycin for 18 days was performed and no further relapse occurred for 32 weeks. In the case of suspected imidocarb resistance in B. canis infection, treatment with atovaquone/azithromycin can be an alternative., Competing Interests: Declaration of competing interest C. Weingart and B. Kohn declare that they have held lectures for veterinary pharmaceutical and diagnostic companies. B. Kohn declares that she has acted as consultant for veterinary pharmaceutical and diagnostic companies and had research collaborations with various companies. J. Krücken declares no conflicts of interest., (Copyright © 2024. Published by Elsevier GmbH.)

In the present study, the effect of a combination therapy consisting of diminazene aceturate (DA) and imidocarb dipropionate (ID) on the in vitro growth of several parasitic piroplasmids, and on Babesia microti in BALB/c mice was evaluated using a fluorescence-based SYBR Green I test. We evaluated the structural similarities between the regularly used antibabesial medications, DA and ID, and the recently found antibabesial drugs, pyronaridine tetraphosphate, atovaquone, and clofazimine, using atom pair fingerprints (APfp). The Chou-Talalay approach was used to determine the interactions between the two drugs. A Celltac MEK-6450 computerized hematology analyzer was used to detect hemolytic anemia every 96 hours in mice infected with B. microti and in those treated with either mono- or combination therapy. According to the APfp results, DA and ID have the most structural similarities (MSS). DA and ID had synergistic and additive interactions against the in vitro growth of Babesia bigemina and Babesia bovis, respectively. Low dosages of DA (6.25 mg kg -1 ) and ID (8.5 mg kg -1 ) in conjunction with each other inhibited B. microti growth by 16.5 %, 32 %, and 4.5 % more than 25 mg kg -1 DA, 6.25 mg kg -1 DA, and 8.5 mg kg -1 ID monotherapies, respectively. In the blood, kidney, heart, and lung tissues of mice treated with DA/ID, the B. microti small subunit rRNA gene was not detected. The obtained findings suggest that DA/ID could be a promising combination therapy for treating bovine babesiosis. Also, such combination may overcome the potential problems of Babesia resistance and host toxicity induced by utilizing full doses of DA and ID., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Background: Vector-borne diseases are of increasing importance in Germany. Since 2015, autochthonous cases have been increasingly documented in Berlin/Brandenburg., Objectives: Describe autochthonous Babesia canis infection in the Berlin/Brandenburg region., Animals: Forty-nine dogs with autochthonous B. canis infection., Methods: Evaluation of history, clinical signs, laboratory abnormalities, treatment, and outcome., Results: Dogs were presented between March and August (9) and September and January (40) in the years 2015-2021. Historical and clinical findings were lethargy (100%), pale mucous membranes (63%), fever (50%), and pigmenturia (52%). Common clinicopathological findings were thrombocytopenia (100%), anemia (85%), intravascular hemolysis (52%), pancytopenia (41%), and systemic inflammatory response syndrome (SIRS; 37%). Babesia detection was based on blood smear evaluation (n = 40) and PCR targeting the 18S rRNA gene of piroplasms (n = 49). Sequencing indicated 99.47% to 100% identity to B. canis sequences from GenBank. All dogs were treated with imidocarb (2.4-6.3 mg/kg; median, 5 mg/kg); 8 dogs received 1, 35 received 2, and 1 dog each received 3, 4, or 5 injections, respectively. Continued PCR-positive results were detected in 7 dogs after the 1st, in 5 after the 2nd, in 2 after the 3rd, and in 1 28 days after the 4th injection. Four dogs were euthanized and 3 dogs died., Conclusions and Clinical Importance: Autochthonous B. canis infections in Berlin/Brandenburg were associated with severe clinicopathological changes, SIRS, and multiorgan involvement. Testing by PCR during and after treatment is advisable to monitor treatment success. Screening of blood donors in high-risk areas and year-round tick protection is strongly recommended., (© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

The expansion of anaplasmosis to non-endemic areas in Argentina has created the need for specific treatments to eliminate Anaplasma marginale from carriers. The most recent studies have failed to chemosterilize A. marginale infections. In this work, we compare the efficacy of long-acting oxytetracycline (OTC) and imidocarb dipropionate (IMD) to chemosterilize the A. marginale infection. For this purpose, twenty steers were randomly clustered into two groups of ten animals each 78 days after A. marginale experimental infection (day 0). Cattle from group 1 (G1) were treated with three doses of 20 mg kg -1 of OTC (Terramycin® LA, 200 mg/ml) 7 days apart by intramuscular injection. Cattle from G2 were treated with two doses of 5 mg kg -1 of IMD (Imizol®, 120 mg/ml) 14 days apart by intramuscular injection. The efficacy of sterilizing treatments was evaluated by detection of DNA by nested PCR, anti-MSP5 antibodies by ELISA and by inoculation of splenectomized calves with blood from the steers 104 days post-treatment (dpt). The results showed 50% efficacy of the OTC treatment to chemosterilize persistent A. marginale infections in cattle and the failure of the IMD treatment under the evaluated conditions. The persistence of specific antibody levels in the sterilized animals (56 dpt) was shorter than the period of DNA detection. The ELISA was the test of choice to confirm the sterilizing outcome after 60 dpt. In spite of its limitations, the sterilization of A. marginale carrier status using OTC, could be useful for high-value bovines in non-endemic areas., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Anaplasma marginale is the causative agent of the severe bovine anaplasmosis. The tick Rhipicephalus microplus is one of the main vectors of A. marginale in tropical and subtropical regions of the world. After the tick bite, the bacterium invades and proliferates within the bovine erythrocytes leading to anemia, impairment of milk production and weight loss. In addition, infection can cause abortion and high mortality in areas of enzootic instability. Immunization with live and inactivated vaccines are employed to control acute bovine anaplasmosis. However, they do not prevent persistent infection. Consequently, infected animals, even if immunized, are still reservoirs of the bacterium and contribute to its dissemination. Antimicrobials are largely employed for the prophylaxis of bovine anaplasmosis. However, they are often used in sublethal doses which may select pre-existing resistant bacteria and induce genetic or phenotypic variations. Therefore, we propose a new standardized in vitro assay to evaluate the susceptibility of A. marginale strains to different antimicrobials. This tool will help health professionals to choose the more adequate treatment for each herd which will prevent the selection and spread of resistant strains. For that, we initially evaluated the antimicrobial susceptibility of two field isolates of A. marginale (Jaboticabal and Palmeira) infecting bovines. The least susceptible strain (Jaboticabal) was used for the standardization of an antimicrobial assay using a culture of Ixodes scapularis-derived tick cell line, ISE6. Results showed that enrofloxacin (ENRO) at 0.25, 1 or 4 μg/mL and oxytetracycline (OTC) at 4 or 16 μg/mL are the most efficient treatments, followed by OTC at 1 μg/mL and imidocarb dipropionate (IMD) at 1 or 4 μg/mL. In addition, this proposed tool has technical advantages compared to the previously established bovine erythrocyte culture. Thereby, it may be used to guide cattle farmers to the correct use of antimicrobials. The choice of the most suitable antimicrobial is essential to eliminate persistent infections, prevent the spread of resistant strains and help controlling of bovine anaplasmosis., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Concerns for recrudescence of Ehrlichia canis infection arise when immunosuppressive drugs are used to treat immune-mediated diseases in dogs previously infected with E. canis., Objectives: Determine whether administration of prednisolone and cyclosporine would reactivate E. canis infection in dogs previously treated with doxycycline during the acute or subclinical phases., Animals: Seven beagles previously experimentally infected with E. canis and administered doxycycline for 4 weeks were included. Three of the 7 dogs were incidentally concurrently infected with Anaplasma platys and Babesia vogeli and were administered 2 doses of imidocarb 2 weeks apart before enrollment in the current study., Methods: Experimental study. Each dog was administered prednisolone and cyclosporine for 6 weeks. Clinical signs, complete blood cell count (CBC), polymerase chain reaction (PCR) assays for E. canis, A. platys, and B. vogeli DNA in blood, E. canis indirect fluorescent antibodies (IFA) titers, and flow cytometry for antiplatelet antibodies were monitored., Results: All dogs completed the immunosuppressive protocol. No evidence for recrudescence of E. canis, A. platys, or B. vogeli were detected based on clinical signs or results of CBC, PCR, IFA, and flow cytometry for antiplatelet antibodies. E. canis IFA titers were negative in 5/7 dogs at the end of immunosuppressive protocol and were negative 6 months after the protocol in 5/5 dogs available for testing., Conclusions and Clinical Importance: Dogs administered with a 4-week course of doxycycline with or without imidocarb failed to show evidence of activation of E. canis infection after administration of a commonly used immune suppressive protocol., (© 2020 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Equine piroplasmosis, caused by the protozoan pathogens Theileria equi or Babesia caballi , is endemic in many (sub-)tropical regions worldwide, including Southern Europe. In Germany, it mainly plays a role as an imported parasitosis. Nevetherless, tick species with known vector potential for equine piroplasms occur in Germany. Thus, a long-term establishment of autochthonous cycles after introduction via infected equids or ticks cannot be excluded. In the present case, 2 horses developed clinical T. equi infection after returning from a trail riding trip to southern France. During treatment, a feeding tick was discovered on one of the horses, which was identified as a Rhipicephalus bursa specimen. This tick species cannot establish in Germany under the current climatic conditions; nevertheless, this discovery illustrates the potential of introducing infected ticks to Germany. The 2 horses were treated with imidocarb (3 mg/kg i. m.; 3 treatments in 3-day intervals) and no adverse drug effects were recorded. To prevent establishment of autochthonous infection cycles in Germany as well as a chronic manifestation of piroplasmosis, treatment should aim at eliminating the parasites despite possible adverse effects of imidocarb., Competing Interests: Die Autoren bestätigen, dass kein Interessenkonflikt besteht., (© Georg Thieme Verlag KG Stuttgart · New York.)

Babesia venatorum was isolated from a captive reindeer calf in Switzerland. The clinical signs consistent with acute babesiosis included hemolytic anemia and hemoglobinuria. The diagnosis was made based on visualization of intraerythrocytic parasites in the stained blood smears and confirmed by PCR analysis of the 18S rRNA gene, with subsequent species identification within Babesia confirmed by sequencing. The reindeer calf was initially treated with supportive care and an antiprotozoal drug (imidocarb dipropionate) but died a few days after hospitalization. Babesia venatorum is also known as Babesia sp. EU1 and can infect different mammalian species, including humans. The current case report aims to increase awareness among veterinarians and reindeer owners about the presence and the associated risk of this zoonotic pathogen. Considering the high morbidity and possible mortality associated with acute babesiosis, captive reindeer should receive tick prevention measures and be tested for subclinical infections in endemic area., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Three different Hepatozoon (Apicomplexa, Hepatozoidae) species have been described infecting domestic cats in Europe (i.e. H. felis, H. canis and H. silvestris), however, reports on clinical hepatozoonosis are uncommon and treatment protocols are not clearly defined. A six-year-old male European short-hair cat from Austria presented poor general condition, lethargy, anorexia, icterus, a painful abdomen, fever, ruffled hair and a tick infestation, and it had never left Austria. Laboratory tests revealed leukopenia, thrombocytopenia and increased serum levels of symmetric dimethylarginine (SDMA) and bilirubin. In May Grünwald-Giemsa-stained blood smears, structures resembling Hepatozoon gamonts were observed inside neutrophil granulocytes. A PCR targeting a fragment of the 18S rRNA gene of Hepatozoon spp. and DNA sequencing allowed the diagnosis of H. felis-DNA in blood samples. The cat was treated with imidocarb dipropionate (6 mg/kg body weight, repeated after 14 days) and doxycycline monohydrate (5 mg/kg body weight twice a day, p.o., for four weeks) and recovered completely. A broad haematological and biochemical laboratory control after six months showed all evaluated parameters under normal ranges. Coinfection with other feline pathogens (i.e. feline leukaemia virus, feline immunodeficiency virus, feline Coronavirus, Leishmania and Dirofilaria immitis) could not be detected. This study reveals the presence of H. felis in Austria and provides more evidence on the geographical distribution and pathogenicity of this parasite for domestic cats. To the authors' knowledge, this is the first autochthonous case of feline hepatozoonosis in Central Europe., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Canine babesiosis (CB) is a significant tick-borne disease caused by intra-erytrocytic protozoa of the genus Babesia (Apicomplexa: Piroplasmida) and is currently emerging or re-emerging in many European countries. Despite the increasing incidence of clinical CB, little is known of the epidemiology of this disease in Romania. Therefore, the present study was conducted to provide epidemiological data on CB in the coastal area of the Black Sea (Dobrogea), southeastern Romania. For this, 306 owned dogs presented during 2016-2018 at a veterinary clinic in the area and clinically suspected for CB were tested for the presence of intra-erytrocytic piroplasms using Giemsa-stained thin blood smears. Overall, 27.8% (95% CI: 22.83-33.16) of dogs were positive for large piroplasms. The Babesia spp.-positive dogs were 51 males and 34 females aged from 3 mo to 13 yr (mean 4.2 yr; SD = 3.6). A higher rate of infection was registered in Bichon Maltese (n = 13) and mixed-breed (n = 28) dogs. Most cases (76/85; 89.4%) were diagnosed from March to June, with only a few cases registered in November and December (1 case each). Infected dogs displayed different clinical presentations, varying from mild (38.8%) to moderate (29.4%) and severe disease (29.4%). Based on clinical- pathological changes, 38 (44.7%), 34 (40.0%), and 13 (15.3%) dogs were diagnosed with uncomplicated babesiosis, complicated babesiosis with a single organ dysfunction, and complicated babesiosis with multiple-organ-dysfunction syndrome (MODS), respectively. The recovery rate (81.6%, 70.1%, and 38.4%, respectively) was significantly different in these groups. The age of dogs appeared to be a risk factor for both severe disease (mean age of 5.8 yr) and MODS (mean age 6.8 yr). These findings document the common occurrence of CB in Dobrogea, southeastern Romania, and suggest that appropriate control measures need to be taken.

Rangeliosis, caused by protozoan Rangelia vitalii, is transmitted by the tick Amblyomma aureolatum. The disease is characterized by hemolytic and hemorrhagic disorder and has been described in dogs and other wild canids. The aim of this study was to compare clinicopathological findings and laboratory results of a Rangelia infection in a crab-eating fox (Cerdocyon thous) with those of canine rangeliosis. The zoo of Universidade de Caxias do Sul, received a crab-eating fox with marked jaundice in mucous membranes, dark-colored stools and neurological signs. The animal underwent an ear tip smear examination and blood collection for complete blood counts, serum biochemistry and PCR. Free-living and intraerythrocytic pyriform structures consistent with R. vitalii were found in the blood smear of the ear tip. The erythrogram revealed normocytic normochromic anemia, moderate macrocytosis, polychromasia and metarubricytosis. The leukogram revealed leukocytosis with neutrophilia and monocytosis, as well as severe thrombocytopenia. Serum biochemistry showed hypoproteinemia, hypoalbuminemia and elevated levels of urea and creatinine. The treatment was performed with imidocarb hydrochloride and dexamethasone, however 24 h after initiation of treatment the animal died. Macroscopic examination revealed jaundice, subcutaneous edema, enlarged superficial lymph nodes, splenomegaly, and hemorrhage of internal organs. Histological sections of the cerebellum, lung, pancreas, intestine and heart were consistent with R. vitalii infection of the vascular endothelium. Pathological and hematological findings were similar to those found in infected dogs, with clinical presentation characterized by hemolytic anemia and hemorrhage. The description of this case showed that C. thous does not only serve as reservoir of R. vitalii but may also develop disease., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

An increasing number of non-endemic vectorborne pathogens have been described in dogs imported to the UK in the past two decades. Recently, an outbreak of canine babesiosis in south-east England has raised veterinary awareness with regard to the impact of such diseases on the UK canine population. Canine hepatozoonosis, caused by Hepatozoon canis and transmitted by the ingestion of Rhipicephalus sanguineus ticks, is widespread in the Mediterranean basin. Herein we describe the first three molecularly confirmed clinical cases of canine hepatozoonosis in dogs imported into the UK. Veterinarians in the UK should be aware of H canis as a potential infection in imported dogs, especially in the face of the expanding distribution of R sanguineus ticks in Europe., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2018. No commercial re-use. See rights and permissions. Published by BMJ.)

Equine piroplasmosis (EP) is a tick-borne disease of equids caused by Babesia caballi and/or Theileria equi, which is endemic in many tropical and temperate areas of the world. However, clinical outbreaks of EP in Romania during the last decades have not been reported Therefore, the aim of this paper is (i) to describe a clinical B. caballi outbreak in horses on several farms in Southern Romania using a diagnostic and therapeutic approach and (ii) the molecular diagnostic of EP in an endemic area of Romania. In the first case, a 10-month-old stallion male was presented with lethargy, anorexia, fever (40.9 °C), pale mucosal/mucous/membranes and a marked anemia. In the subsequent weeks, three horses from other farms located in the same area, displayed similar clinical signs. B. caballi was diagnosed in all the horses based on Giemsa-stained blood smears and the diagnosis was further confirmed by polymerase chain reaction (PCR), using a single-round and multiplex PCR and sequencing. All four horses were treated with imidocarb dipropionate, at a dose rate of 2.2 mg/kg body weight (two injections at 48 h apart), and all horses clinically recovered within 24-48 h, post-treatment. This report presents the first molecularly characterized B. caballi outbreak in Romania in clinically affected horses, confirmed by DNA sequencing.

Certain countries including the United States remain non-endemic for particular infectious diseases such as equine piroplasmosis through import restrictions and surveillance. Endemic regions often employ premunition as the primary method to control disease, however in non-endemic countries, chemosterilization combined with methods to confirm parasite elimination are required to maintain disease-free status. The ability of imidocarb diproprionate (ID) to clear persistent Theileria equi infection from infected horses has been shown through the inability of treated horses to transmit via blood transfer. However, the common lengthy persistence of anti-T. equi antibody causes regulatory tests such as cELISA or IFA to remain positive for extended periods. Persistence of positive testing creates challenges for regulatory veterinary medicine and international trade. Concordance between nested polymerase chain reaction (nPCR) targeting the ema1 gene and immunoblotting (IB) measuring declination in anti-EMA1 and anti-EMA2 antibody were used to verify clearance of T. equi from 179 ID-treated horses. These data support the use of IB to demonstrate declining anti-EMA1 and EMA2 titers in T. equi-infected horses subsequent to successful ID treatment. Such data provide concordant support to a negative nPCR and allow for a more timely determination of effective ID clearance of T. equi. The post ID treatment results indicate that while nPCR was consistently negative by 14 days and cELISA generally remained positive after 1 year, immunoblot was on average negative after 4 months and 100% in agreement with nPCR., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Background: Classification of Babesia parasites has traditionally relied on morphological differentiation based on piroplasm size and shape. Molecular typing has subsequently revealed a more complex taxonomy for these piroplasms than previously thought. To evaluate the factors that influence the morphology of Babesia species upon microscopic examination and hence, their taxonomic classification, we performed detailed characterizations of piroplasms from archival and prospective collections of cytological samples of dogs with piroplasmosis before and after death. Merozoite morphology and time of parasite disappearance following imidocarb dipropionate was also investigated., Methods: The study was divided into a (i) review of archived cytological slides from confirmed cases of canine piroplasmosis, and (ii) a prospective study of smears and tissue imprints from 15 recently necropsied dogs. The latter group could be further sub-divided into a non-treated group and an imidocarb dipropionate-treated group. Exact times of treatment before death were reviewed. Additional blood smears prepared from the live dogs and taken before therapy were also evaluated in the latter group. Parasite burden per each slide was determined in both studies. The shape and size of merozoites were described from blood smears taken while the dogs were alive and from different organs during necropsy. The results of all measurements were statistically analyzed., Results: The morphology and size of merozoites from live dogs corresponded to that of previously described 'large' Babesia. The morphology and size of merozoites were significantly different (P < 0.001) in postmortem samples, however, and more consistent in shape and size with piroplasm cells previously referred to as 'small' Babesia. PCR and sequencing confirmed B. canis as the causative agent of disease in all investigated dogs, including in postmortem negative tissue imprints from dogs treated at least 24 h before death., Conclusions: Changes in the morphology of 'large' B. canis to 'small'-like Babesia observed by light microscopy appear to represent a common postmortem change. Classification of Babesia parasites into 'large' and 'small' Babesia using only microscopy of postmortem slides should be treated with caution. PCR-based methodologies for detection and molecular typing of Babesia spp. may prove valuable for investigating suspected cases of babesiosis following necropsy.

Background: Feline cytauxzoonosis is an emerging infection caused by tick-transmitted apicomplexan parasites of the genus Cytauxzoon. The association of clinical disease with Cytauxzoon infection appears to be limited to C. felis infections in the Americas. Sporadic infections of wild and domestic felids with Cytauxzoon sp. were recently described in European countries but clinical reports of the infection are rare and incomplete. This case report brings new interesting information on cytauxzoonosis expression in Europe., Case Presentation: A 9-years-old castrated European shorthair cat living in rural area of north-eastern France (Saint Sauveur, Bourgogne-Franche-Comté region), without any travel history was presented for consultation due to hyperthermia, anorexia, depression and prolonged fever that didn't respond to antibiotic therapy. The cat had outdoor access with a history of vagrancy and was adequately vaccinated (core vaccines and FeLV vaccine). During biological investigations, intraerythrocytic inclusions were observed on blood smear and were further investigated by PCR analysis and sequencing. Molecular analyses confirmed Cytauxzoon sp. infection. The cat was treated with a subcutaneous injection of imidocarb dipropionate (3.5 mg/kg). One week after treatment, the cat improved clinically, although parasitic inclusions within erythrocytes persisted, and only a mild lymphocytosis was found. Two weeks after treatment, the cat appeared in excellent health, appetite was normal and parasitemia was negative. However, one month after treatment the cat relapsed with hyperthermia, anorexia, and depression. Blood smears and PCR were once again positive. Subsequently, the cat received an additional dose of imidocarb dipropionate (3.5 mg/kg SC) and recovered rapidly without other clinical signs. Two weeks after the second imidocarb injection, the cat was hit by a car and died., Conclusion: This case provides the first clinical description of infection by Cytauxzoon sp. in a domestic cat in France. These findings support the fact that cytauxzoonosis should be considered in the differential diagnosis of acute febrile illness which does not respond to antibiotic in cats with outdoor access especially in areas where populations of wild felids are present.

Background: Piroplasmosis caused by the Babesia microti-like piroplasm (Bml) is increasingly being detected in dogs in Europe. Sick dogs show acute disease with severe anaemia associated with thrombocytopenia with a poor response to current available drugs. This study assesses the safety and tolerance of three treatments and compares their efficacy over a full year of follow up in dogs naturally infected with Bml., Methods: Fifty-nine dogs naturally infected with Bml were randomly assigned to a treatment group: imidocarb dipropionate (5 mg/kg SC, 2 doses 14 d apart) (IMI); atovaquone (13.3 mg/kg PO q 8 h, 10 d)/azithromycin (10 mg/kg PO q 24 h, 10 d) (ATO); or buparvaquone (5 mg/kg IM, 2 d apart)/azithromycin (same dosage) (BUP). Before and after treatment (days 15, 45, 90 and 360), all dogs underwent a physical exam, blood tests and parasite detection (blood cytology and PCR). Clinical efficacy was assessed by grading 24 clinical and 8 clinicopathological signs from low to high severity., Results: Before treatment, most dogs had severe regenerative anaemia (88.13%) and thrombocytopenia (71.4%). On treatment Day 45, clinical signs were mostly reduced in all dogs, and by Day 90, practically all dogs under the ATO or BUP regimen were clinically healthy (76.4 and 88%, respectively). Highest percentage reductions in laboratory abnormalities (82.04%) were detected in animals treated with ATO. Over the year, clinical relapse of Bml was observed in 8 dogs (8/17) treated with IMI. However, on Day 360, these animals had recovered clinically, though clinicopathological abnormalities were still present in some of them. Parasitaemia was PCR-confirmed on Days 90 and 360 in 47.05 and 50% of dogs treated with ATO, 68 and 60.08% with BUP, and 94.1 and 73.3% with IMI, respectively. Even after 360 days, 13.3% of the dogs treated with IMI returned a positive blood cytology result., Conclusions: IMI showed the worse clinical and parasitological, efficacy such that its use to treat Bml infection in dogs is not recommended. The treatments ATO and BUP showed better efficacy, though they were still incapable to completely eliminate PCR-proven infection at the recommended dose. All three treatments showed good tolerance and safety with scarce adverse events observed.

Background: The apicomplexan hemoparasite Theileria equi is a causative agent of equine piroplasmosis, eradicated from the United States in 1988. However, recent outbreaks have sparked renewed interest in treatment options for infected horses. Imidocarb dipropionate is the current drug of choice, however variation in clinical response to therapy has been observed., Methods: We quantified the in vitro susceptibility of two T. equi isolates and a lab generated variant to both imidocarb dipropionate and a bumped kinase inhibitor compound 1294. We also evaluated the capacity of in vitro imidocarb dipropionate exposure to decrease susceptibility to that drug. The efficacy of imidocarb dipropionate for clearing infection in four T. equi infected ponies was also assessed., Results: We observed an almost four-fold difference in imidocarb dipropionate susceptibility between two distinct isolates of T. equi. Four ponies infected with the less susceptible USDA Florida strain failed to clear the parasite despite two rounds of treatment. Importantly, a further 15-fold decrease in susceptibility was produced in this strain by continuous in vitro imidocarb dipropionate exposure. Despite a demonstrated difference in imidocarb dipropionate susceptibility, there was no difference in the susceptibility of two T. equi isolates to bumped kinase inhibitor 1294., Conclusions: The observed variation in imidocarb dipropionate susceptibility, further reduction in susceptibility caused by drug exposure in vitro, and failure to clear T. equi infection in vivo, raises concern for the emergence of drug resistance in clinical cases undergoing treatment. Bumped kinase inhibitors may be effective as alternative drugs for the treatment of resistant T. equi parasites.

Objective: The purpose of this study was to track changes in selected subpopulations of lymphocytes in the blood of dogs infected with Babesia (B.) canis and treated with imidocarb., Material and Methods: The study included 16 dogs divided into two groups. The first group (n = 6) consisted of healthy control animals. Dogs of the se- cond group (n = 10) were infected with B. canis and after establishment of the diagnosis each animal received a single dose of imido- carb (5 mg/kg). Flow cytometry was used to enumerate several immune cell phenotypes., Results: It was concluded that the invasion of B. canis contributes to the decreased percentage of CD3+, CD4+, CD8+, CD21+ lymphocytes in the blood of infected animals. The decreased level of tested subpopulations of lymphocytes in group 2 persisted for the entire 12-day period of the test. After the administration of imidocarb, each tested lymphocyte fraction in the blood of the dogs with babesiosis increased, but did not reach physiological values., Conclusion: The presented results indicate that the resolution of clinical signs associated with babesiosis may be related to the stimulation and intensity of cellular immunity, dependent on the CD4+ T cells profile. After administration of imidocarb, the parasitemia is cleared which allows the recovery of the lymphocyte populations.

Paraoxonase 1 (PON1) is a negative acute phase protein bound to high density lipoproteins (HDL) and during the acute phase response (APR) protects HDL from peroxidation. The aim of this study was to assess the relationship between PON1 and HDL in canine babesiosis, a disease characterized by oxidative damages and by an APR. PON1, HDL and C-reactive protein (CRP), were measured in blood collected from 15 controls and 29 dogs with babesiosis sampled at admission, and on days 1 and 7 after treatment. At admission, PON1 and HDL were significantly lower in affected dogs. HDL concentration increased at day 1 while PON1 increased and CRP decreased at day 7. This suggests that the decrease of PON1 at admission is in part due to an increased consumption, the decreased HDL may depend on lipid peroxidation and its rapid increase after treatment may depend on the antioxidant activity of PON1., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We determined the prevalence of tick-borne pathogens in domestic dogs using microscopy and polymerase chain reaction (PCR) techniques. A total of 303 EDTA blood samples were collected from domestic dogs in Khon Kaen Province, Thailand, in May 2013. Microscopic observation of Giemsa-stained smears and molecular diagnosis using conventional PCR were performed. Infected dogs were treated with imidocarb dipropionate, a combination of imidocarb dipropionate and doxycycline, or doxycycline alone. Seventy-one (23.4%) out of 303 dogs were positive for DNA of tick-borne pathogens. Of the 303 animals, 13.2% and 1.3% were positive for a single infection with Babesia spp or Ehrlichia canis, respec- tively using microscopy; whereas 19.5% and 3.0% were positive using the PCR technique. Co-infection with Babesia spp and E. canis was observed in 0.7%, and coinfection with Hepatozoon canis and E. canis in 0.3%. Infected dogs were treated with the assigned drugs, and elimination of the pathogens was demonstrated by microscopy and PCR. The results indicated that while both microscopic and PCR diagnostic techniques were useful for tick-borne pathogen detection, PCR was more effective. Imidocarb dipropionate and doxycycline were found to be effective for treatment of babesiosis and ehrlichiosis, respectively. The present study suggests that the PCR technique has high sensitivity and specificity for Babesia and Ehrlichia diagnosis as well as for detection of Babesia spp, E. canis and H. canis DNA in EDTA blood specimens.

Theileriosis is a tick-borne disease caused by a piroplasma of the genus Theileria that can causeanaemia and thrombocytopenia. Its clinical importance for dogs' remains poorly understood,as only some develop clinical signs. In this study, physical and laboratory findings, treatment and outcomes of six client-owned diseased dogs presented at the Onderstepoort Veterinary Academic Hospital are described retrospectively. In the dogs, Theileria species (n = 4) and Theileria equi (n = 2) were detected by a polymerase chain reaction (PCR)-reverse blothybridisation assay in blood samples, whilst PCR for Babesia, Anaplasma and Ehrlichia were negative. The most common physical findings were pale mucous membranes (five out of six dogs), bleeding tendencies (five out of six dogs) and lethargy (three out of six dogs). All dogs were thrombocytopenic [median 59.5 x 10(9)/L (range 13-199)] and five out of six dogs were anaemic [median haematocrit 18% (range 5-32)]. Bone marrow core biopsies performed in two dogs showed myelofibrosis. Theileriosis was treated with imidocarb dipropionate and the suspected secondary immune-mediated haematological disorders with prednisolone and azathioprine. Five dogs achieved clinical cure and post-treatment PCR performed in three out of five dogs confirmed absence of circulating parasitaemia. An immune-mediated response to Theileria species is thought to result in anaemia and/or thrombocytopenia in diseased dogs with theileriosis. A bleeding tendency, most likely secondary to thrombocytopenia and/or thrombocytopathy, was the most significant clinical finding in these cases. The link between thrombocytopenia, anaemia and myelofibrosis in theileriosis requires further investigation and theileriosis should be considered a differential diagnosis for dogs presenting with anaemia and/or thrombocytopenia in endemic tick-borne disease areas.

Hepatozoonosis caused by Hepatozoon canis (Eucoccidiorida, Hepatozoidae) is among the most widespread vector-borne infections of dogs, primarily transmitted by Rhipicephalus sanguineus sensu lato ticks. Based on the absence of a consensus on the treatment regimes for canine hepatozoonosis, the present study aimed to evaluate the efficacy of imidocarb dipropionate (5-6 mg/kg subcutaneously once a week for 6 weeks), and of toltrazuril/emodepside (Procox(®), 15 mg/kg once a day for 6 days) in association with clindamycin (15 mg/kg once a day for 21 days) in treating naturally infected dogs. At the enrollment time (T0), 32 dogs, cytologically or molecularly positive for H. canis, were assigned to test and control groups. Animals were treated according to the specific therapeutic protocol, and the presence of H. canis gamonts was assessed weekly by cytology and PCR throughout six months (T1-T19). In addition, any abnormality in leucocyte morphology was evaluated and recorded. Results indicate that, in spite of a reduction in the percentage of infected dogs, both treatments did not provide parasitological cure. Accordingly, new treatment protocols or active compounds against H. canis should be investigated., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In the present study, the treatment of ichthyophthiriasis with medicated feed was investigated in rainbow trout, Oncorhynchus mykiss, and chub, Leuciscus cephalus. The anti-parasitics toltrazuril and imidocarb; the antibiotics doxycycline, erythromycin and sulphadiazine and the anti-inflammatory acetylsalicylic acid were tested. In vitro experiment revealed that all tested anti-parasitics and antibiotics were effective in killing the isolated trophonts and theronts. Minimum doses for killing 100 % of the viable trophonts and for inhibiting the development of theronts were 3 mg/L for doxycycline, 30 mg/L for erythromycin, 2 mg/L for imidocarb dipropionate, 30 mg/L for sulphadiazine and 20 mg/L for toltrazuril. Acetylsalicylic acid (40 mg/kg fish/day), doxycycline (3 and 6 mg/kg/day), erythromycin (40 mg/kg/day), imidocarb dipropionate (5.0 mg/kg/day), sulphadiazine (40 mg/kg/day), toltrazuril (20 and 40 mg/kg/day) and combinations of doxycycline and toltrazuril (3 + 20 mg/kg/day, 6 + 40 mg/kg/day) were tested as medicated feed. When administered as medicated feed, only doxycycline, toltrazuril and combinations of doxycycline and toltrazuril reduced the fish mortality and infestation level. Best results were obtained by feeding a combination of 6 mg/kg/day doxycycline and 40 mg/kg/day toltrazuril. In O. mykiss, this treatment reduced the mortality rate from 100 to 50 ± 14 % after 10 days and the infestation level from grade 4 (≥100 trophonts per skin mucus sample) to 3.5 (50-100 trophonts). In L. cephalus, the mortality rate was decreased from 100 to 39 ± 5 % and the infestation level from grades 4 to 2 (ten to 50 trophonts) after 10 days.

Acute tubular necrosis (ATN) was described in canine babesiosis. Hypotension is considered as one of the factors which influence the development of hypoxic renal damage. In this study hypotension defined as mean arterial pressure (MAP) < 80 mmHg was detected in 7 out of 48 dogs (14.6%) infected with Babesia canis. Lower systolic arterial pressure (SAP), diastolic arterial pressure (DAP) and MAP were detected in azotaemic dogs infected with B. canis. Statistically significant negative correlations between blood pressures (SAP, DAP and MAP) and serum creatinine and urea concentrations showed the influence of decreased blood pressure on the development of azotaemia and is probably also associated with ATN in canine babesiosis.

Two young brother male free-ranging domestic shorthair cats were evaluated for diarrhea. They presented with intraerythrocytic piroplasms on blood smear evaluation. Only the first cat was anemic (mild non-regenerative anemia). A partial segment of the 18S rRNA was amplified and sequenced, revealing a homology of 99% with Cytauxzoon sp. and of 93% with Cytauxzoon felis. The first cat was treated with doxycycline and imidocarb dipropionate and monitored by serial laboratory exams, resulting negative for Cytauxzoon sp. infection after the end of the therapy (follow-up period of 175 days). The second cat received the same therapy, but doxycycline was discontinued by the owner after 1 week. He was monitored for 130 days, remaining erythroparasitemic and asymptomatic. We described cases of Cytauxzoon sp. infection in domestic cats with detailed clinical data, description of two therapeutic protocols, and follow-up after treatment with opposite parasitological responses (parasitological cure versus persistence of infection).

A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles., (© 2013 American Society for Veterinary Clinical Pathology.)

Background: Babesia infections in dogs can result in a wide range of clinical and laboratory presentations, including coagulopathy. Expression of intercellular adhesion molecule-1 (ICAM-1) and von Willebrand factor (vWF) in dogs with babesiosis is unknown., Objectives: Whether inflammation in babesiosis triggers activation of ICAM-1 and the coagulation system., Animals: Twelve and 10 dogs with naturally occurring babesiosis before and after antiparasitic treatment, respectively, were compared with 10 healthy dogs., Methods: In this prospective study, diagnosis was made by blood smear examination and confirmed by PCR. C-reactive protein (CRP), soluble intercellular adhesion molecule 1 (sICAM-1), and von Willebrand factor (vWF) levels were measured by a canine ELISA kit, fibrinogen (FIB) and factor VIII activity levels were measured by coagulometric methods, and blood cell counts (WBC, RBC, PLT) were determined with an automatic analyzer., Results: Compared to healthy dogs, the CRP, sICAM-1, and FIB concentrations were significantly increased before therapy and remained high for 3 days after therapy in dogs with babesiosis. vWF activity was significantly decreased in dogs with babesiosis before treatment. FVIII activity did not differ between dogs with babesiosis and healthy dogs. WBC; RBC and PLT were significantly lower before treatment and normalized by 3 days after treatment., Conclusion and Clinical Importance: A proinflammatory condition in babesiosis appears to influence endothelial dysfunction and hemostatic activity. Although clearly beneficial for the parasite, sequestered blood cells can obstruct blood flow in small vessels, promote an inflammatory state, and could increase the severity of babesiosis., (Copyright © 2013 by the American College of Veterinary Internal Medicine.)

Anaplasma platys was first identified and described in North America as a Rickettsia-like, platelet-specific organism in dogs with infectious canine cyclic thrombocytopenia. In Europe, A. platys has so far mainly been described for some Mediterranean countries. Here, we describe a case of A. platys infection in a dog from Romania, confirmed by PCR. Additionally, the dog had a co-infection with Hepatozoon canis. To the best of our knowledge, this is the first case of A. platys infection in Romania and the first case of a co-infection with A. platys and H. canis altogether. Both pathogens should be considered as possible disease agents in dogs suffering from disease associated with tick bite in south-eastern Europe., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Theileria equi, one of the causative agents of equine piroplasmosis, is endemic in many regions of the world but is considered a 'foreign' animal disease in the USA. In an effort to prevent the importation of T. equi, stringent serological screening of horses is practiced prior to entry to the USA. Current regulatory options available where horses are found to be infected include permanent quarantine with or without chemotherapy, repatriation, or euthanasia. Chemotherapeutics that eliminate infection and subsequently transmission risk are critical in the management of infected horses. In this study, the efficacy of the drug imidocarb dipropionate against experimental T. equi infection was assessed. Of nine horses experimentally inoculated with T. equi isolated from an animal previously imported from Peru, six were treated with imidocarb dipropionate after the resolution of the acute phase of the disease. Elimination of the parasite was demonstrated in 5/6 by nested PCR, blood transfusions to naïve horses, and reversion to seronegative status. The findings support the use of this drug as a potential treatment option in controlling outbreaks of T. equi, and also suggest that 'combination testing' using both serological and PCR detection methods are necessary to demonstrate clearance of infection., (Published by Elsevier Ltd.)

Ovine babesiosis, caused by Babesia ovis, is of major economic importance in Turkey. The changes in the blood profile of infected animals are informative about the course of infection. The aim of the present study was to evaluate the hematological and biochemical changes in the pre- and post-treatment periods of the natural B. ovis infections. The presence of the parasites was confirmed by microscopy and polymerase chain reaction (PCR) analysis. On the basis of the clinical and laboratory findings, the infections were categorized into different groups according to the degree of anemia and the level of parasitemia. All infected sheep were treated with imidocarb dipropionate (IMDP). The blood pictures in the pre- and post-treatment periods were compared. Pancytopenia occurred in animals with severe anemia and very high parasitemia, and bicytopenia in the other groups. The platelet count (PLT), plateletcrit (PCT) and mean platelet volume (MPV) returned to the normal ranges after treatment, except those in the group with severe anemia. In the biochemical profile, B. ovis infection caused an increase in blood urea nitrogen and total bilirubin, and these parameters returned to normal levels after treatment. The indirect fluorescein antibody test (IFAT) results showed that 38.1% of the cases raised specific antibodies during the period of infection, with titers ranging from 1/160 to 1/640. All of 45 animals re-examined after treatment were seropositive, with high titers that rose up to 1/5120., (Copyright © 2012 Elsevier B.V. All rights reserved.)

An 8-yr-old, captive, spayed, female maned wolf (Chrysocyon brachyurus) developed progressive lethargy and weakness over a 24-hr period. Clinical signs included vomiting, recumbency, horizontal nystagmus, possible blindness, pale icteric mucus membranes, and port-wine colored urine. A complete blood cell count revealed severe anemia (packed cell volume [PCV], 6%) and intraerythrocytic piroplasms consistent with a Babesia species. Polymerase chain reaction testing later confirmed babesiosis. The wolf was treated with imidocarb dipropionate, antibiotics, and fluid therapy. A whole-blood transfusion from a sibling maned wolf also was performed. Despite aggressive treatment, the wolf failed to improve and was euthanized. To the authors' knowledge, this is the first documented case of babesiosis in a captive maned wolf in North America. Surveillance of infectious diseases in captive and wild maned wolf populations should be expanded to include screening for Babesia species. Tick control also should be implemented to prevent and decrease transmission of the disease to this endangered species.

Arthropod-borne apicomplexan pathogens that cause asymptomatic persistent infections present a significant challenge due to their life-long transmission potential. Although anti-microbials have been used to ameliorate acute disease in animals and humans, chemotherapeutic efficacy for apicomplexan pathogen elimination from a persistently infected host and removal of transmission risk is largely unconfirmed. The recent re-emergence of the apicomplexan Theileria equi in U.S. horses prompted testing whether imidocarb dipropionate was able to eliminate T. equi from naturally infected horses and remove transmission risk. Following imidocarb treatment, levels of T. equi declined from a mean of 10(4.9) organisms/ml of blood to undetectable by nested PCR in 24 of 25 naturally infected horses. Further, blood transfer from treated horses that became nested PCR negative failed to transmit to naïve splenectomized horses. Although these results were consistent with elimination of infection in 24 of 25 horses, T. equi-specific antibodies persisted in the majority of imidocarb treated horses. Imidocarb treatment was unsuccessful in one horse which remained infected as measured by nested PCR and retained the ability to infect a naïve recipient via intravenous blood transfer. However, a second round of treatment eliminated T. equi infection. These results support the utility of imidocarb chemotherapy for assistance in the control and eradication of this tick-borne pathogen. Successful imidocarb dipropionate treatment of persistently infected horses provides a tool to aid the global equine industry by removing transmission risk associated with infection and facilitating international movement of equids between endemic and non-endemic regions.

A 5-month-old intact male Boerboel dog, imported from South Africa 1 week previously, was presented to a Texas veterinarian for lethargy, anorexia, and labored breathing. The dog was febrile, anemic, leukopenic, thrombocytopenic, and slightly azotemic. Results of the IDEXX SNAP-4Dx enzyme immunoassay were negative for Dirofilaria immitis antigen and antibodies against Ehrlichia canis, Borrelia burgdorferi, and Anaplasma phagocytophilum. An EDTA blood sample analyzed at Oklahoma State University Center for Veterinary Health Sciences revealed nonregenerative anemia, neutropenia, and large protozoal piroplasms in 0.7% of the RBCs. Piroplasms were 2-5μm long and varied in shape from round to oval to piriform; extracellular merozoites were also observed. Nested PCR was performed on DNA extracted from blood using primers that amplify the 18s rRNA gene from all known Babesia species, and the product was sequenced. Basic Local Alignment Search Tool analysis of the 437 base sequence revealed 99-100% similarity to Babesia canis rossi, 92-93% similarity to Babesia canis canis, and 92% similarity to Babesia canis vogeli. The dog responded well to treatment with imidocarb. PCR analysis of a second blood sample 2 weeks later was negative for Babesia spp. DNA. This case represents the first diagnosis of B. canis rossi infection in the United States., (©2011 American Society for Veterinary Clinical Pathology.)

In December 2007, Babesia bovis was introduced to New Caledonia through the importation of cattle that had been vaccinated with a live tick fever (babesiosis and anaplasmosis) vaccine. Although the tick Rhipicephalus (Boophilus) microplus is common in New Caledonia, the territory had previously been free of tick-borne diseases of cattle. This paper describes the initial extent of the outbreak, the measures and rationale for disease control, and the progress to date of the eradication campaign. Initially, 22 properties were affected involving approximately 2300 cattle in 'high risk' zones and 1600 in adjoining 'suspect' zones. Rather than slaughtering infected herds or attempting to eliminate the tick vector, the campaign was based on quarantine of affected properties, and aggressive tick control in conjunction with 3-monthly treatments of the high risk cattle with the antiprotozoal drug imidocarb dipropionate. Subsequent surveillance by ELISA and PCR showed a progressive and dramatic decline in seroprevalence among infected herds and the absence of new infections. All 22 properties were considered to be free of Babesia within 12 months of the start of the disease control program. These results indicate that the strategy was effective in eliminating Babesia from infected herds and feasible as an eradication strategy on a moderately large scale. Unfortunately, early in the campaign, babesiosis spread to a herd of feral cattle on a property in the 'suspect' zone, and this reservoir of infection subsequently resulted in the infection (or reinfection) of cattle on several neighbouring commercial farms. The eradication campaign in New Caledonia is currently focussed on destocking the feral cattle - extensive surveillance suggests that this is the only remaining nidus of infection., (2010 Elsevier GmbH. All rights reserved.)

Background: Imidocarb or a combination of atovaquone and azithromycin (A&A) has been suggested for treatment of cats with cytauxzoonosis, but neither has been prospectively evaluated for efficacy., Hypothesis/objectives: That survival to hospital discharge is improved by treatment with A&A as compared with imidocarb., Animals: Eighty acutely ill cats with Cytauxzoon felis infection treated at one of 18 veterinary clinics in 5 states., Methods: An open-label, randomized prospective study compared survival in cats treated with atovaquone (15 mg/kg p.o. q8h) and azithromycin (10 mg/kg p.o. q24h) or imidocarb (3.5 mg/kg i.m.). All received heparin, fluids, and supportive care. Clinical and clinicopathologic data from initial presentation were collated. Parasitemia was quantified (n = 79) and pathogens genotyped (n = 60). Logistic regression was used to determine the impact of treatment group on the primary endpoint, survival to hospital discharge or death. Covariants were analyzed by rank-sum testing., Results: Of 53 cats treated with A&A, 32 (60%) survived to discharge while only 7 of 27 cats (26%) treated with imidocarb survived (P = .0036; odds ratio 7.2, 95% CI 2.2, 24). Cats with a lower parasitemia were more likely to survive, as were cats with higher white blood cell counts and lower total bilirubin. Unique pathogen genotypes were identified from 15 cats, while genotype isolated from 21 cats had been described previously. There were multiple pathogen genotypes identified in 24 cats., Conclusions and Clinical Importance: Survival to discharge was more likely in cats treated with A&A as compared with imidocarb, although case fatality rate remained high., (Copyright © 2010 by the American College of Veterinary Internal Medicine.)

The efficacy of imidocarb dipropionate for the treatment of Hepatozoon canis infection was studied in three naturally infected asymptomatic dogs followed longitudinally over 8 months. Response to treatment was followed by monitoring blood counts, parasitemia levels in blood, parasite in concentrated buffy-coat smears and by PCR. The dogs were initially treated with a low dose of 3 mg/kg imidocarb dipropionate twice a month and when parasitemia persisted after five treatments, with the regular dose of 6 mg/kg. In one dog, H. canis gamonts were no longer detectable by blood and buffy-coat microscopy after 2 months of therapy with 6 mg/kg while in the two other dogs gamonts were intermittently found in blood but persistently detectable in buffy-coat smears during the whole study period. Furthermore, combined therapy with doxycycline monohydrate administered at 10 mg/kg/day PO for 4 weeks also failed to eliminate H. canis. PCR revealed that parasite DNA was present in the blood of all dogs at all sampling dates regardless of treatment refuting the effectiveness of treatment suggested by negative blood microscopy. Detection of H. canis in buffy coat was found to be twice as sensitive than by blood smear and detection by PCR was even more sensitive revealing infection in eight samples (16% of total samples) negative by blood and buffy-coat microscopy. In conclusion, imidocarb dipropionate was not effective in eliminating H. canis from dogs treated repeatedly over 8 months. Microscopical detection is not sufficient for the evaluation of treatment response in H. canis infection and follow up by molecular techniques is recommended., (Copyright 2010 Elsevier B.V. All rights reserved.)

An Irish setter from the Oslo area was presented to the clinic with signs of babesiosis, a few days after a tick bite. Blood analysis confirmed babesiosis. Microscopic examination of thin blood film revealed large, basophilic, bodies inside erythrocytes, indicative of a large Babesia sp. Molecular analysis using PCR, indicated the presence of a Babesia spp. in the blood. Sequencing and phylogenetic analysis of the PCR fragment revealed a sequence which was 100% identical to Babesia canis canis 18S. As this dog had never been abroad, it can be concluded that this is the first report of an autochthonous infection of B. canis canis in Norway., (Copyright 2010 Elsevier B.V. All rights reserved.)

A 10-year-old tame zebra gelding was presented after suffering from lethargy, nervousness, reported anaemia and icterus as well as a decreased appetite. These symptoms were seen over some months, with changing severity. The animal was immobilised, treated, and blood specimens were submitted for haematology and biochemistry. This report describes molecular characterisation of Theileria equi recovered from this animal, as well as the clinical findings, treatment and historical relevance of piroplasmosis in zebra in southern Africa.

Case Description: A 12-year-old 46-kg (101.2-lb) sexually intact male Labrador Retriever was evaluated because of lymphadenomegaly. The dog resided in Texas, and its travel history included many southeastern and eastern shore states but not North Carolina., Clinical Findings: Following evaluation of the dog, a diagnosis of stage IVa intermediate- to large-cell lymphoma was made. A cyclophosphamide-hydroxydaunorubicin (doxorubicin)-vincristine-prednisone chemotherapy protocol was initiated. One week after the first chemotherapeutic treatment, a routine blood smear evaluation revealed single and paired intraerythrocytic large piroplasms that resembled Babesia canis. Via molecular testing, the organism was identified as a Babesia sp that had been detected previously in dogs in North Carolina., Treatment and Outcome: The dog was administered imidocarb diproprionate (7 mg/kg [3.2 mg/lb], IM) on 2 occasions (3-week interval). At 1, 4, 15, and 50 weeks after the second treatment, blood samples were analyzed specifically for the North Carolina Babesia sp via PCR assay; the result of each assay was positive., Clinical Relevance: Because of the morphologic similarity of the large piroplasm detected in dogs in North Carolina to B canis, molecular testing of large piroplasms detected in dogs is needed to definitively identify the infective Babesia sp. In the dog of this report, the infection was not eliminated following treatment with imidocarb diproprionate, which may have been a result of the immunocompromised state of the dog or the drug's ineffectiveness against this parasite. If imidocarb diproprionate is ineffective against the North Carolina Babesia sp, treated dogs may act as reservoirs of infection.

Antimicrobial treatment of persistent infection to eliminate transmission risk represents a specific challenge requiring compelling evidence of complete pathogen clearance. The limited repertoire of antimicrobial agents targeted at protozoal parasites magnifies this challenge. Using Babesia caballi as both a model and a specific apicomplexan pathogen for which evidence of the elimination of transmission risk is required for international animal movement, we tested whether a high-dose regimen of imidocarb dipropionate cleared infection from persistently infected asymptomatic horses and/or eliminated transmission risk. Clearance with elimination of transmission risk was supported by the following four specific lines of evidence: (i) inability to detect parasites by quantitative PCR and nested PCR amplification, (ii) conversion from seropositive to seronegative status, (iii) inability to transmit infection by direct inoculation of blood into susceptible recipient horses, and (iv) inability to transmit infection by ticks acquisition fed on the treated horses and subsequently transmission fed on susceptible horses. In contrast, untreated horses remained infected and capable of transmitting B. caballi using the same criteria. These findings establish that imidocarb dipropionate treatment clears B. caballi infection with confirmation of lack of transmission risk either by direct blood transfer or a high tick burden. Importantly, the treated horses revert to seronegative status according to the international standard for serologic testing and would be permitted to move between countries where the pathogen is endemic and countries that are free of the pathogen.

Changes in coagulation parameters were studied in dogs naturally infected with Babesia canis canis (n = 30), and haemostasis was evaluated and compared to values obtained from healthy dogs (n = 29). To date, there have not been any studies examining the dynamics of thrombin-antithrombin complex formation in cases of canine babesiosis. Coagulation parameters evaluated before (day 0) and on days 1, 2, and 3 after treatment with imidocarb (6 mg/kg inj. s.c.) included the determination of platelet counts, the formation of thrombin-antithrombin complexes (TAT), prothrombin time (PT), activated partial thromboplastin time (APTT) and antithrombin III (AT III) activity. TAT complexes were significantly elevated in animals with babesiosis on days 0 and 2 (mean 49.7 and 87.7 microg/L vs. control, 7.2 microg/L). AT III activity was significantly decreased at all time-points examined. There were no differences in PT. On days 2 and 3 the APTT was significantly shortened in the infected dogs when compared to control animals (means of 21.3 and 19.2 s vs. control, 30.0 s). Our analysis demonstrated that infected dogs had significant thrombocytopenia during the course of the study (mean day 0 - 29 x 10(9) /L, day 1 - 48 x 10(9) /L, day 2 - 47 x 10(9) /L and day 3 - 87 x 10(9) /L, vs. control -259 x 10(9) /L). These data suggest that babesiosis in dogs compromise primary and secondary haemostasis and that induction of disseminated intravascular coagulation (DIC) occurs in canine babesiosis.

Two captive reindeer (Rangifer tarandus tarandus) at a New York zoological institution were diagnosed with Babesia odocoilei. Clinical signs consistent with acute babesiosis included fever, hemoglobinuria, and hemolytic anemia. Both episodes were precipitated by stressful events that may have compromised their immunocompetence. The diagnosis was confirmed by visualization of intraerythrocytic parasites on stained blood smears, polymerase chain reaction, and speciation of the Babesia by sequencing a hypervariable region of the 18S rRNA gene. One reindeer died with gross and histopathologic lesions, including pigmentary nephrosis with severe acute tubular degeneration and necrosis secondary to intravascular hemolysis. A second reindeer was successfully treated with supportive care and an antiprotozoal, imidocarb dipropionate (Imizol, 12%, Schering-Plough Animal Health, Union, New Jersey 07083, USA) at 3 mg/kg s.c. or i.m. s.i.d. on days 1, 2, 6, 9, and 21. Two other reindeer in the exhibit tested negative for Babesia by polymerase chain reaction but were treated with imidocarb dipropionate as prophylaxis while final testing results were pending. Additionally, B. odocoilei was identified in three novel asymptomatic host species within the collection: yak (Bos grunniens), muntjac (Muntiacus reevesi), and markhor goat (Caprafalconeri). Due to the high morbidity and mortality associated with acute babesiosis, captive reindeer should receive tick prevention, be tested for subclinical infections in endemic areas, and receive aggressive treatment for acute infections when clinical babesiosis is suspected.

The aim of the study was to trace the clinical course of babesiosis in 76 dogs infected with Babesia canis protozoa and to assess the usefulness of PCR method in the routine diagnosis of the disease. The investigations were conducted in three successive seasons of the biological activity of ticks on dogs displaying possible clinical signs of babesiosis, the latter assigned individual numbers from 001 to 076. All the animals underwent routine clinical examinations and blood was collected for haematological, biochemical, parasitological and molecular tests for babesiosis. The most frequent clinical signs observed in the course of the disease were changes in urine colour and xanthosis or paleness of mucous membranes, whereas in the haematological and biochemical examinations, the most frequent laboratory findings were thrombocytopenia, leucopoenia, anaemia and an increase in levels of urea and bilirubin. In all blood smears stained with the May-Grunwald and Giemsa methods, from the 76 dogs, the presence of Babesia canis protozoa was observed in erythrocytes, and their DNA was detected in 69 blood samples by means of PCR technique. The course of the disease and the results of molecular examinations suggested the haemolytic form of babesiosis. The previous genetic analysis of isolates of Babesia canis canis from the eastern areas of Poland helped to distinguish two specific groups, A and B, within the species (Adaszek and Winiarczyk 2008a). The present study revealed a certain interrelation between the intensification of thrombocytopenia and the fact that protozoa belong to either group A or B. The mean number of thrombocytes in dogs infected with protozoa from group A was 61.11 thousand/mm3, whereas the mean number of thrombocytes in the blood of dogs infected with protozoa from group B was 27.47 thousand/mm3. A strong correlation was also observed between the low level of thrombocytes and the increase in the internal body temperature (p = 0.02), accelerated pulse rate (p = 0.01) and discoloration of urine (p = 0.04). As a result of the treatment of dogs with imidocarb, recovery was observed in 73 out of the 76 dogs brought to the clinic.

Repeated polymerase chain reaction (PCR) testing of 3 asymptomatic domestic cats were positive for Cytauxzoon felis DNA, suggesting persistent infection. Two cats initially presented with clinical signs consistent with acute cytauxzoonosis and, in both cases, signs of illness resolved after treatment. Parasitemia was detected in peripheral blood smears from these cats upon presentation with illness and, at subsequent follow-up appointments, in the absence of clinical illness. Polymerase chain reaction analysis was positive for C. felis from blood sampled at each time point. A third cat, a housemate of a cat fatally infected with C. felis, was preventatively treated for infection at the time of the housemate cat's death. This contact cat, having never shown signs of clinical illness consistent with cytauxzoonosis infection, had no detectable parasitemia but was positive for C. felis on repeated PCR testing. Detection of asymptomatically infected cats allows for the possibility of a yet unrecognized population of infected domestic cats that may have the capacity to serve as an additional reservoir host for C. felis, altering the currently accepted paradigm of C. felis transmission to domestic cats through bobcats as the reservoir host. In cases of very low parasitemia, more sensitive means of parasite detection, such as PCR testing, may be necessary to detect infected cats. Increased detection of asymptomatically infected cats will aid in understanding the epidemiology of C. felis infection and enhance the ability to prevent this highly fatal infectious disease of domestic cats.

An Akita Inu, living in Belgium, was presented with unusual clinical manifestations of acute babesiosis that included neurological signs and pancytopenia. Diagnosis was made by identifying Babesia canis in the blood smear. Neurological signs resolved after treatment with imidocarb diproprionate. Normalization of hematological abnormalities was gradual over 5 months.

Imidocarb treatment of horses infected with Babesia caballi is supposed to eliminate the infection, but data on the efficacy of this treatment is scarce. The study presented here concerns four Paso Fino horses, which were imported into the island of Curacao on the basis of a piroplasmosis negative complement fixation test (CFT). Upon re-testing with an indirect fluorescent antibody test immediately after arrival in Curacao, two horses appeared to have antibodies to B. caballi and all horses had antibodies to Theileria equi. Subsequent testing with polymerase chain reaction combined with a reverse line blot yielded positive results for both agents in all four horses. Treatment with five consecutive doses of imidocarb dipropionate (4.7 mg/kg BW im q 72 h), temporarily resulted in negative results, but B. caballi and T. equi were detected again in the samples taken at 6 and 18 weeks after completion of the treatment. These results confirm that the CFT is not a suitable test for pre-import testing and that even high dose treatment with imidocarb may not be capable of eliminating B. caballi and T. equi infections from healthy carriers.

Babesiosis (piroplasmosis) is a tick-borne disease with a symptoms of hemolytic anemia. For the first time babesiosis was described in dogs in United States in 1934. The etiological factor of this disease in Poland is protozoa Babesia canis, and its vector--Dermacentor-tick. The most common symptoms of babesiosis are: icterus, hemoglobinuria, occasionally vomits and diarrhea. The biochemical examination of blood serum from sick animals can reveal the increase of activity of AST, ALT, the increase of total bilirubine, urea and creatynine concentrations. The results of hematological examinations can show anemia, leucopenia and thrombocytopenia. The diagnosis of babesiosis bases on anamnesis, clinical examinations of dogs, microscopical examinations of blood smears from sick animals; IF-assay and PCR can also be helpful for the diagnosis of babesiosis. Till now does not exist the effective immunoprophylaxis against this disease. Babesiosis is well-known disease, however there are still problems with therapy of infected animals. Most effective drug in therapy of dog piroplasmosis is imidocarb, but sometimes can be observed side effects after it application. It is possible that the genetically differences which are detected in subspecies may have an influence on the severity of disease and the effectiveness of therapy.