Canis Vogeli Research Articles

Surveillance of tick-borne pathogens in domestic dogs from Chad, Africa

BackgroundTick-borne pathogens are understudied among domestic animals in sub-Saharan Africa but represent significant threats to the health of domestic animals and humans. Specifically, additional data are needed on tick-borne pathogens in Chad, Africa. Surveillance was conducted among domestic dogs in Chad for selected tick-borne pathogens to measure (1) the prevalence of antibodies against Anaplasma spp., Borrelia burgdorferi, and Ehrlichia spp.; (2) the prevalence of infections caused by Hepatozoon spp., Ehrlichia canis, Anaplasma platys, and Babesia spp.; and (3) associations of pathogens with demographic, spatial, and temporal factors. Blood samples were collected from domestic dogs at three time points (May 2019, November 2019, June 2020) across 23 villages in southern Chad.ResultsOf the 428 dogs tested with the IDEXX SNAP 4Dx test in May 2019, 86% (n = 370, 95% CI = 83–90%) were positive for antibodies to Ehrlichia spp., 21% (n = 88, 95% CI = 17–25%) were positive for antibodies to Anaplasma spp., and 0.7% (n = 3, 95% CI = 0.1–2%) were positive for antibodies to Borrelia burgdorferi. Four different pathogens were detected via PCR. Hepatozoon spp. were most commonly detected (67.2–93.4%, depending on the time point of sampling), followed by E. canis (7.0-27.8%), A. platys (10.1–22.0%), and Babesia vogeli (0.4–1.9%). Dogs were coinfected with up to three pathogens at a single time point, and coinfections were most common in May 2019 compared to November 2019 and May 2020.ConclusionsOverall, this study provides new data about the epidemiology of tick-borne pathogens in domestic dogs in Chad, with potential implications for dog and human health.

Molecular detection and characterization of Anaplasma marginale and Babesia canis vogeli infecting dogs in Luxor, Egypt

Tick-borne diseases in animals are increasing rapidly worldwide, but there is insufficient information about tick-borne diseases infecting dogs in southern Egypt. Thus, in the current study, we detected the presence of Anaplasma marginale (A. marginale) and Babesia canis vogeli (B. canis vogeli) in the blood of dogs. The results revealed that 4/100 (4%) were positive, and a higher infection rate was found in males (75%), than females (25%). The phylogenetic analysis for the major surface protein 4 (msp4) gene in this study was compared with amplicons separate from other reported isolates with alignment by identity 100% with cattle and camels from Egypt, and the phylogenetic analysis for the B. canis vogeli small subunit ribosomal RNA (SSU rRNA) gene in this study identified identity by 99.89% with dogs from Egypt. This report is considered the first report in southern Egypt about A. marginale in dogs based on the sequence analysis of the msp4 gene, providing new data for the classification and identification of A. marginale in dogs compared to A. marginale isolated from other animals in southern Egypt.

Molecular screening and genetic diversity of tick-borne pathogens associated with dogs and livestock ticks in Egypt.

The Middle East and North Africa (MENA) offer optimal climatic conditions for tick reproduction and dispersal. Research on tick-borne pathogens in this region is scarce. Despite recent advances in the characterization and taxonomic explanation of various tick-borne illnesses affecting animals in Egypt, no comprehensive examination of TBP (tick-borne pathogen) statuses has been performed. Therefore, the present study aims to detect the prevalence of pathogens harbored by ticks in Egypt. A four-year PCR-based study was conducted to detect a wide range of tick-borne pathogens (TBPs) harbored by three economically important tick species in Egypt. Approximately 86.7% (902/1,040) of the investigated Hyalomma dromedarii ticks from camels were found positive with Candidatus Anaplasma camelii (18.8%), Ehrlichia ruminantium (16.5%), Rickettsia africae (12.6%), Theileria annulata (11.9%), Mycoplasma arginini (9.9%), Borrelia burgdorferi (7.7%), Spiroplasma-like endosymbiont (4.0%), Hepatozoon canis (2.4%), Coxiella burnetii (1.6%) and Leishmania infantum (1.3%). Double co-infections were recorded in 3.0% (27/902) of Hy. dromedarii ticks, triple co-infections (simultaneous infection of the tick by three pathogen species) were found in 9.6% (87/902) of Hy. dromedarii ticks, whereas multiple co-infections (simultaneous infection of the tick by ≥ four pathogen species) comprised 12% (108/902). Out of 1,435 investigated Rhipicephalus rutilus ticks collected from dogs and sheep, 816 (56.9%) ticks harbored Babesia canis vogeli (17.1%), Rickettsia conorii (16.2%), Ehrlichia canis (15.4%), H. canis (13.6%), Bo. burgdorferi (9.7%), L. infantum (8.4%), C. burnetii (7.3%) and Trypanosoma evansi (6.6%) in dogs, and 242 (16.9%) ticks harbored Theileria lestoquardi (21.6%), Theileria ovis (20.0%) and Eh. ruminantium (0.3%) in sheep. Double, triple, and multiple co-infections represented 11% (90/816), 7.6% (62/816), and 10.3% (84/816), respectively in Rh. rutilus from dogs, whereas double and triple co-infections represented 30.2% (73/242) and 2.1% (5/242), respectively in Rh. rutilus from sheep. Approximately 92.5% (1,355/1,465) of Rhipicephalus annulatus ticks of cattle carried a burden of Anaplasma marginale (21.3%), Babesia bigemina (18.2%), Babesia bovis (14.0%), Borrelia theleri (12.8%), R. africae (12.4%), Th. annulata (8.7%), Bo. burgdorferi (2.7%), and Eh. ruminantium (2.5%). Double, triple, and multiple co-infections represented 1.8% (25/1,355), 11.5% (156/1,355), and 12.9% (175/1,355), respectively. The detected pathogens' sequences had 98.76-100% similarity to the available database with genetic divergence ranged between 0.0001 to 0.0009% to closest sequences from other African, Asian, and European countries. Phylogenetic analysis revealed close similarities between the detected pathogens and other isolates mostly from African and Asian countries. Continuous PCR-detection of pathogens transmitted by ticks is necessary to overcome the consequences of these infection to the hosts. More restrictions should be applied from the Egyptian authorities on animal importations to limit the emergence and re-emergence of tick-borne pathogens in the country. This is the first in-depth investigation of TBPs in Egypt.

Molecular detection of Babesia spp. in dogs in Germany (2007–2020) and identification of potential risk factors for infection

BackgroundIn Europe, canine babesiosis is most frequently caused by Babesia canis and Babesia vogeli, and occasionally by Babesia gibsoni.. In Germany, B. canis is recognized as endemic. The aims of this study were to assess how often Babesia spp. infections were diagnosed in a commercial laboratory in samples from dogs from Germany, and to evaluate potential risk factors for infection.MethodsThe database of the LABOKLIN laboratory was screened for Babesia spp.-positive polymerase chain reaction (PCR) tests for dogs for the period January 2007–December 2020. Sequencing was performed for positive tests from 2018 and 2019. Binary logistic regression analysis was performed to determine the effects of sex, season, and year of testing. Questionnaires were sent to the submitting veterinarians to obtain information on travel abroad, tick infestation, and ectoparasite prophylaxis of the respective dogs. Fisher’s exact test was used to calculate statistical significance and P < 0.05 was considered statistically significant.ResultsIn total, 659 out of 20,914 dogs (3.2%) tested positive for Babesia spp. by PCR. Of 172 sequenced samples, B. canis was identified in 156, B. vogeli in nine, B. gibsoni in five, and B. vulpes in two. Season had a statistically significant impact on test results when summer/winter (1.6% tested positive) was compared to spring/autumn (4.7%), with peaks in April (5.2%) and October (7.4%) [P < 0.001, odds ratio (OR) = 3.16]. Sex (male 3.5%, female 2.8%; P = 0.012, OR = 1.49) and age (< 7 years old 4.0%, ≥ 7 years old 2.3%; P < 0.001, OR = 1.76) of the tested dogs also had a statistically significant effect. A statistically significant impact was demonstrated for observed tick attachment (P < 0.001, OR = 7.62) and lack of ectoparasite prophylaxis (P = 0.001, OR = 3.03). The frequency of positive Babesia spp. tests did not significantly differ between the 659 dogs that had never left Germany and the 1506 dogs with known stays abroad (P = 0.088).ConclusionsThe possibility of canine infection with B. canis needs to be especially taken into consideration in spring and autumn in Germany as the activity of the tick Dermacentor reticulatus, a potential vector for canine babesiosis, is highest in these seasons. Travel and importation of dogs are considered major factors associated with canine babesiosis in Germany. However, autochthonous Babesia spp. infections also occur in a considerable number of dogs in Germany.Graphical

Molecular characterization of babesia species infecting dogs (Canis familiaris) in the Federal Capital Territory, Abuja, Nigeria

Babesiosis is a tick-borne disease caused by an intra-erythrocytic protozoan of the genus Babesia. The genus contains a large number of species capable of infecting a variety of mammalian hosts, including humans. Misdiagnosis of the infection is a major setback in disease management, monitoring, and control. Therefore, timely and precise identification of infecting Babesia species is critical. This study aimed at detecting and characterizing the Babesia species infecting dogs in the Federal Capital Territory (FCT) Abuja. Blood samples were collected from apparently healthy dogs (n=480), thin smears were prepared, Giemsa-stained, and examined under a microscope. DNA was extracted from positive blood samples (n=15), and PCR amplification of the 18S rRNA gene was performed using generic primers. PCR products were purified, electrophoresed on 1.5% agarose gels and thereafter, sequenced bidirectionally. The prevalence of Babesia species in thin smear was 3.1%. The prevalence in the six Area Councils was 5.0%, 3.75%, 2.5%, 2.5%, 2.5% and 2.5% for Gwagwalada, Municipal, Bwari, Kuje, Kwali and Abaji respectively. The prevalence in relation to age was 3.87% in dogs &gt;1year old and 1.76% in dogs ≤ 1 year old. Prevalence by breed, was 4.38% in exotic dogs, 3.23% in local and 2.35% in crossbred. The prevalence in relation to sex was 3.96% in males and 2.37% in females. The BLAST search was carried out in the NCBI database, and a phylogenetic tree was constructed using nucleotide sequences in GenBank. Electrophoresed PCR products revealed a band size of about 612bp. Blast in NCBI demonstrated 100% similarity with Babesia canis vogeli nucleotide sequences. This study reaffirmed the molecular identification of B. c. vogeli in apparently healthy dogs in Nigeria and it may be of clinical importance to veterinary clinicians in the management and control of the disease.

Multiple blood pathogen infections in apparently healthy sheltered dogs in southern Thailand

ABSTRACT In developing countries such as Thailand, free-ranging dogs are frequently involved in road accidents and contribute to the cost of public healthcare. Shelters play a vital role in communities because they help to control the population of unwanted and free-ranging dogs. This study aimed to investigate blood pathogen infection in sheltered dogs, as it is one of the factors contributing to animal welfare. Blood samples were randomly collected from 141 dogs from the largest shelter (approximately 400–500 dogs in total) in southern Thailand. Blood pathogens were detected using both PCR and light microscopy. Four blood pathogens were identified: Anaplasma platys, Ehrlichia canis, Babesia canis vogeli, and Hepatozoon canis. No trypanosomes were detected. The incidence of blood parasite infection was 56.7% (80/141) by PCR, and 28.4% (40/141) by microscopy. E. canis was the most prevalent pathogen, accounting for 46.1% (65/141) of the cases, while multiple infections accounted for 22% (31/141) of the cases. A triple infection with E. canis, A. platys, and B. canis vogeli was observed in 5.7% (8/141) of the cases. Although PCR is far more sensitive than microscopy, it appears to have equivalent specificity. In conclusion, this study reported a high occurrence of blood pathogen infections in clinically healthy sheltered dogs. Many of them were infected with multiple pathogens and may have been infected before entering the shelter. These findings suggest that a blood test is necessary to screen dogs prior to their admission to the shelter to prevent disease transmission and enhance animal welfare.

Screening and phylogenetic characterization of tick-borne pathogens in a population of dogs and associated ticks in Egypt

BackgroundThe incidence or recurrence of tick-borne diseases (TBDs) in animals and humans is increasing rapidly worldwide, but there is insufficient information about TBDs infecting dogs in Egypt. Thus, the present study was conducted to screen and genetically identify tick-borne pathogens (TBPs) in dogs and associated ticks by microscopic examination and polymerase chain reaction (PCR).MethodsIn Cairo and Giza governorates, 208 blood samples were collected from dogs of different breeds, ages, and sex. In addition, 1266 dog-associated ticks were collected (546 ticks were used to prepare hemolymph smears, and 720 ticks were kept in 70% ethanol until PCR analysis). PCR was applied to 124 dog blood samples and 144 tick pools prepared from 720 ticks.ResultsAll ticks collected from dogs were Rhipicephalus sanguineus (s.l.). Microscopic examination revealed that TBP prevalence among dogs was 23.56% (49/208), including Anaplasma and Ehrlichia with 11.1% (23/208) and Babesia canis with 8.2% (17/208). Hepatozoon canis was not detected in blood smears. Co-infections with two pathogens were visible in 4.33% (9/208) of examined dogs. The prevalence of TBPs in hemolymph smears was 45.97% (251/546) including 35.89% (196/546) for H. canis, 8.1% (44/546) for B. canis, and 2.01% (11/546) for Anaplasmataceae (A. phagocytophilum, A. marginale, A. platys, and E. canis). The overall molecular prevalence rate of TBPs was 25.81% and 29.17% in the blood of examined dogs and in ticks, respectively. The molecular prevalence of Anaplasmataceae family, Babesia canis, and H. canis in dog blood samples was 19.35%, 6.45%, and 0.0%, respectively, while in ticks, it was 20.83%, 5.55%, and 2.8%, respectively. A sequential analysis identified six different species of TBPs, namely B. canis vogeli, Hepatozoon canis, A. phagocytophilum, A. marginale, A. platys, and E. canis. The obtained sequences were submitted to GenBank and assigned accession numbers.ConclusionsThe present study detected a wide range of TBPs (B. canis, H. canis, A. platys, A. phagocytophilum, A. marginale, and E. canis) that are considered a threat to domestic animals and humans in Egypt. Hepatozoon canis and A. marginale were reported in dogs and associated ticks for the first time in Egypt.Graphical

Molecular detection and control of non- regenerative anaemia associated with Babesia gibsoni and Anaplasma platys co- infection in a dog

Anaemia and thrombocytopenia are the common clinical features of blood parasite infection in dogs. Blood parasites previously identified in dogs in Kerala were Babesia gibsoni, Babesia canis vogeli, Ehrlichia canis and Trypanosoma evansi. A two-year old female Labrador Retriever was brought to the University Veterinary Hospital, Mannuthy, Thrissur, with a complaint of anorexia for the past two weeks. A thorough clinical examination did not give any evidence of involvement of other body systems, except for a splenomegaly which was confirmed by radiography. The dog was negative for microfilaria and intestinal parasites. Haematological findings revealed moderate anaemia and thrombocytopenia. Anaemia was classified as non-regenerative based on reticulocyte count. Blood smear was positive for basophilic intra-erythrocytic inclusions suggestive of small babesia and basophilic inclusions in platelets suggestive of morula of Anaplasma spp. Species-specific PCR on whole blood genomic DNA-showed specific amplification for B. gibsoni and A. platys organisms and negative for B. canis vogeli, E. canis and T. evansi. The present study reports a case of non-regenerative anaemia due to B. gibsoni and A. platys co-infection in a dog and its successful therapeutic management. This is the second report of A. platys infection in dogs in Kerala, India.

Diagnóstico molecular para hemoparasitos em cães atendidos no Hospital Veterinário da Universidade Santo Amaro, São Paulo, SP, Brasil

OBJECTIVE: Detection of Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, Rangelia vitalii, and Babesia canis vogeli using real-time PCR in dogs treated at the veterinary hospital of the Universidade Santo Amaro, located in the south zone, in the city of São Paulo, SP, Brazil. METHODS: DNA was extracted from 63 whole blood samples using the “PureLink Genomic DNA” extraction kit (Invitrogen®) according to the manufacturer's instructions and real-time PCR was performed to detect Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, Rangelia vitalii, and Babesia canis vogeli. RESULTS: In total, 23.8% (15/63) of the samples were positive by real-time PCR for at least one pathogen. Of these, 9.52% (6/63) were positive for Babesia canis vogeli and 14.2% (9/63) for Ehrlichia canis. No samples were positive for Rickettsia rickettsii, Rangelia vitalii, and Anaplasma platys. CONCLUSION: This study demonstrated, in an unprecedented way, the presence of B. canis vogeli and E. canis in dogs from fragmentation areas of the Atlantic Forest around the Guarapiranga Reservoir in the city of São Paulo, expanding knowledge on the dispersion of this agent in Brazil.

Clinical, Hematological, Blood Gasometric and Electrolytic Changes in Dogs Receiving Whole Blood Transfusions

Background: Blood transfusion is a tool capable of saving lives. Patients undergoing blood transfusion usually present several alterations in the acid-base and electrolyte balance, aggravating the condition of critically ill patients. Some studies have demonstrated haematological alterations in certain species that received whole blood transfusions, however, few studies have evaluated acid base and electrolyte changes in dogs undergoing whole blood haemotherapy. The aim of this study was to analyze clinical, hematological, blood gas and electrolyte changes in anemic dogs after whole blood transfusion.Materials, Methods &amp; Results:Twenty nine dogs transfused due to anemia were enrolled in the study. Donors blood was collected in a transfusion bag containing citrate phosphate adenine dextrose and stored up to 24 h. Blood collections and evaluations were made before and 24 h after the transfusion. Data distribution normality was tested by the Shapiro Wilk Test. The means of the variables were compared by paired t-test. It was observed an increase in diastolic blood pressure and a reduction in heart rate (P &lt; 0.05). There was a not significant increase in systolic blood pressure, temperature, and a reduction in respiratory rate per minute. Erythrocyte, haemoglobin and haematocrit averages were significantly increased after blood transfusion (P &lt; 0.05). It was observed a reduction in the mean values of pH (P &lt; 0.05), potassium (P &gt; 0.05) and ionized calcium (P &gt; 0.05) and an increase in the mean partial pressure of carbon dioxide (pCO2) (P &lt; 0.001), bicarbonate (P &gt; 0.05) and sodium (P &lt; 0.05). The probable cause of anemia was monocytic ehrlichiosis (14/29), visceral leishmaniasis (1/29), babesiosis (1/29), co-infection of Ehrlichia canis and Leishmaniainfantum (2/19), co-infection of E. canis and Babesia vogeli (1/19). It was not possible to determine the etiology of the anemia in ten dogs. Discussion:Heart rate significantly reduced after transfusion, probably because of the increase in hematocrit, hemoglobin and erythrocyte values. It may be justified by the displacement of extravascular fluid to the intravascular space. Mean values of systolic blood pressure were slightly elevated before transfusion and remained elevated afterwards, while diastolic and mean arterial pressure increased significantly after transfusion. These changes may be due to the morbid condition and may be influenced by many other factors. Haematocrit, haemoglobin and erythrocyte values increased significantly after transfusion, according to what was observed in other studies. The significant reduction in pH and increase in pCO2 reflects the compensatory mechanism for metabolic acidosis to increase ventilation, leading to pCO2 reduction and changes in pH. The reduction in pH due to the contact of the collected blood with conservative solutions is one of the main changes thar occurs during blood storage. It was described significantly lower pH in dogs’ whole blood samples stored for more than 24h in vacutainer plastic containing CPDA-1. We may assume there was no intense pH reduction in the present study because the bags were stored for up to 24 h. Although not statistically significant, the increase of pO2 mean reflects the improvement of tissue oxygen perfusion. It was observed a significant increase in sodium ions. The mean sodium ionconcentration before transfusion was very close to the maximum reference value. Hyperkalaemia was not observed, nor was there significant reduction of potassium ions after transfusion. Several studies report hyperkalaemia and transfusion-associated cardiac arrests in humans, associated with infusion of large volumes of blood. Whole blood transfusion increased erythrogram values and did not negatively affect the electrolyte or acid-base status, representing a safe and useful tool in the intensive care of small animals.

Euthyroid Sick Syndrome and Changes in Thyroid Hormones in Dogs with Hemoparasitosis

Background: Hemoparasitoses are extremely important in the clinical routine because they affect a large number of dogs. In spite of the abundance of studies on this topic, hormonal alterations caused by infection with these agents are still poorly known. Therefore, the goal of this work was to assess the serum levels of thyroid hormones of dogs infected with Ehrlichia canis (E. canis) alone, and dogs infected with E. canis and Babesia canis vogeli (B. vogeli) and/or Anaplasma platys (A. platys) before and after treatment with doxycycline chlorohydrate. This study also aimed at checking for presence of euthyroid sick syndrome (ESS) in these animals.Materials, Methods &amp; Results: The concentrations of the thyroid hormones total triiodothyronine (TT3), total tetraiodothyronine (TT4), free tetraiodothyronine (FT4), and canine thyroid-stimulating hormone (cTSH) were assessed by chemiluminescence in 12 dogs. Nested polymerase chain reaction (nPCR) was used to confirm diagnoses. The dogs were divided into 2 groups: G1, which comprised animals infected by E. canis alone, and G2, which included animals simultaneously infected by E. canis and B. vogeli and/or A. platys. The serum concentrations of the thyroid hormones were measured at two time points: before (D1) and after (D2) the 28-day treatment with generic doxycycline chlorohydrate (DC) at a dose of 10 mg/kg SID. On D2, another nPCR was carried out to check the efficacy of the treatment. On D2, in both groups, all dogs became negative for E. canis; however, 8 animals remained infected or were reinfected by other hemoparasites. On D1, 4 dogs in G1 exhibited low TT3 in conjunction with low TT4; one of the dogs had increased TT3 alone, and another dog had an increased TT3 accompanied by decreased TT4. In G2, on D1, one dog exhibited high TT3 accompanied by a decreased concentration of TT4; 2 dogs had decreased TT4; 2 dogs had increased TT3; and one dog had both TT3 and TT4 decreased. On D2, with only one exception, the animals in G1 that became negative for all hemoparasites under study exhibited normal serum levels of TT3 and TT4. Three dogs in G1, which were reinfected on D2, sustained decreased TT4 serum concentrations. Whereas, in G1 on D2, only one dog remained with a reduced serum TT4 concentration.Discussion: DC was effective at eliminating infection by E. canis in all dogs on D2. Even though this drug was unable to prevent reinfection of the animals by other hemoparasites, and in spite of divergent results on its efficacy at eliminating babesiosis and anaplasmosis, a clinical improvement was observed in all patients. A decrease in the serum concentrations of thyroid hormones (ESS) may occur in dogs with severe non-thyroid related diseases. Different authors have demonstrated the presence of ESS in dogs with hemoparasitoses; however, this is the first study to provide evidence on the existence of this syndrome in dogs with hemoparasitoses in Brazil, including dogs simultaneously infected by E. canis and B. vogeli and/or A. platys.We observed a reduction in TT3 and/or TT4 in dogs infected by E. canis, B. vogeli, and A. platys. We believe these results may help assessment of prognosis and patient follow-up, as TT4 concentrations returned to normal levels in all dogs that became negative for all hemoparasites after treatment; and only one dog in G1 sustained increased TT3, albeit lower than observed on D1. In animals of G1 that were reinfected after treatment started, TT3 returned to normal, even though TT4 levels remained decreased. In sum, all dogs in G1 and G2 exhibited TT3 and/or TT4 concentrations outside of the reference value ranges on D1, which indicates that alterations in the levels of thyroid hormones were due to infection by the hemoparasites. The animals infected by E. canis alone or simultaneously infected by B. vogeli and/or A. platys exhibited ESS.

Molecular detection of &lt;i&gt;Babesia canis vogeli&lt;/i&gt; and &lt;i&gt;Hepatozoon canis&lt;/i&gt; in dogs in the department of Magdalena (Colombia)

The canine population in the cities of Ciénaga and Santa Marta has been estimated at 54,953 based on individual dogs with owners. Due to the role that dogs play in society, either as pets or as transmitters of zoonoses to humans, we conducted a study with 169 blood samples from dogs that visited two veterinary clinics in these locations between March and September of 2017. The objective of the study was to detect species of Babesia and Hepatozoon canis by amplifying the 18S gene using conventional polymerase chain reaction (PCRc). The presence of Babesia sp. and Hepatozoon canis was detected in 15 (8.87%) and 12 (7.10%) DNA samples, respectively. In addition, 7 (4.14%) cases of coinfection were recorded. The Babesia sp. sequences obtained corresponded to the B. canis vogeli subspecies. This both pathogens in the Colombian Caribbean region and cases of coinfection in Colombian dogs. Therefore, the national veterinary community is encouraged to consider the information presented here in their differential diagnoses associated with companion vector-borne diseases (CVBDs). This information will allow veterinary professionalsto create control and prevention strategies to prevent the spread of these infections.

Hepatozoon canis and Babesia vogeli infections of dogs in Tunisia.

A paucity of studies is available on haemoparasites in dogs in Tunisia. In this study, we used molecular techniques (PCR/sequencing) to detect and characterize haemoprotozoa in sick dogs from Tunisia. A total of 99 dogs displaying such clinical symptoms as fever, anorexia, and depression were presented for treatment to the hospital of the Veterinary School of Sidi Thabet (Tunisia). Among dogs screened by PCR, five (5%) proved to be infected with a hemoprotozoa species. An analysis of all the sequences that were obtained enabled us to identify two species of Protozoa: Babesia vogeli (in three dogs) and Hepatozoon canis (in two other dogs). This is the first time that an infection of dogs by Hepatozoon canis in Tunisia has been reported. Veterinary practitioners should be aware that these two haemoparasites can infect dogs and should include them in any differential diagnosis of clinical illnesses with manifestations compatible with tick-borne diseases.

Babesia canis spp. in dogs in Baghdad Province, Iraq: First molecular identification and clinical and epidemiological study.

Aims:The aim of this study was to investigate babesiosis in dogs of different breeds and ages and of both sexes in Baghdad Province by molecular detection of Babesia canis using conventional polymerase chain reaction (PCR) and sequencing followed by phylogenetic analyses.Materials and Methods:Blood samples were collected from 310 dogs of different ages and breeds, and of both sexes in different areas of Baghdad Province from December 2018 to September 2019; during clinical examinations, body temperature, pulse, respiratory rate, and signs of diseases were recorded. PCR was used to amplify a specific 450-bp fragment of the 18S rRNA gene of B. canis. PCR products were sequenced, and MEGA 6.0 software was used for analysis. Chi-square and odds ratio tests were used to investigate the prevalence and risk factors of babesiosis.Results:Clinical signs of babesiosis included paleness or icterus of the mucus membranes, tick infestation, and febrile illness during the acute and subacute phase. The prevalence of infection with B. canis was 5.1%, with the higher prevalence in male dogs and in dogs <3 years of age. Huskies were more likely to be infected than other dogs. Infection prevalence was highest in April and June and was higher in spring and summer than in winter. Using sequence data, 14 isolates of Babesia canis canis and one isolate of each Babesia canis rossi and Babesia canis vogeli were identified. Phylogenetic analyses of B. canis canis revealed that three shared clades and several isolated lineages were similar to other isolates (97-99% similarity), whereas B. canis vogeli and B. canis rossi showed similarities of 98% and 99% with isolates from other geographical regions.Conclusion:This study provides the first molecular record and phylogenic analysis of B. canis in dogs in Iraq, and it will be valuable for confirming clinical signs and studying epidemiological risk factors of babesiosis in dogs.

A novel metabarcoding diagnostic tool to explore protozoan haemoparasite diversity in mammals: a proof-of-concept study using canines from the tropics

Haemoparasites are responsible for some of the most prevalent and debilitating canine illnesses across the globe, whilst also posing a significant zoonotic risk to humankind. Nowhere are the effects of such parasites more pronounced than in developing countries in the tropics where the abundance and diversity of ectoparasites that transmit these pathogens reaches its zenith. Here we describe the use of a novel next-generation sequencing (NGS) metabarcoding based approach to screen for a range of blood-borne apicomplexan and kinetoplastid parasites from populations of temple dogs in Bangkok, Thailand. Our methodology elucidated high rates of Hepatozoon canis and Babesia vogeli infection, whilst also being able to characterise co-infections. In addition, our approach was confirmed to be more sensitive than conventional endpoint PCR diagnostic methods. Two kinetoplastid infections were also detected, including one by Trypanosoma evansi, a pathogen that is rarely screened for in dogs and another by Parabodo caudatus, a poorly documented organism that has been previously reported inhabiting the urinary tract of a dog with haematuria. Such results demonstrate the power of NGS methodologies to unearth rare and unusual pathogens, especially in regions of the world where limited information on canine vector-borne haemoparasites exist.

Ameliorative effects of alpha-lipoic acid and imidocarb dipropionate on clinico-haematological changes induced by experimental Babesia canis vogeli infection in dogs

The aim of the study was to evaluate the effects of imidocarb dipropionate and alpha-lipoic acid (ALA) on clinico-haematological parameters of dogs inoculated with Babesia canis vogeli. Twenty-one dogs were divided into uninfected untreated (n = 4), infected untreated (n = 6), infected and treated with imidocarb dipropionate (6 mg/kg subcutaneously) alone (n = 3), infected and treated with ALA (15 mg/kg orally) alone (n = 5), and infected and treated with both imidocarb dipropionate and ALA (n = 3). Molecularly identified Babesia canis vogeli was experimentally inoculated into the infected groups, and treatment commenced at day 12 post-inoculation (PI). Behavioural events and blood samples were obtained and analysed before inoculation, at days 11 PI, 8, 15, and 43 post-treatment (PT). Behavioural events including vocalisation, licking/grooming, and tail wagging decreased post-inoculation. Treatments with imidocarb and ALA alleviated the clinical manifestations of dry muzzle, ocular discharge, and splenomegaly. At days 15 and 43 PT, imidocarb + ALA recorded the highest packed cell volume (41.13 ± 2.96%; 47.40 ± 3.72%) and erythrocyte count (6.10 ± 0.44 × 1012/L; 6.99 ± 0.56 × 1012/L) compared to other infected groups. The haemoglobin concentration at day 15 PT (13.80 ± 0.91 g/dL) increased in the imidocarb + ALA group. Thrombocytopenia was recorded in all the infected groups at day 11 PI. In conclusion, co-administration of imidocarb with ALA acted synergistically and also ameliorated the clinico-haematological changes in experimental canine babesiosis, and the agent may be a promising and potent drug in the management of canine babesiosis.

Molecular detection of Babesia canis vogeli in dogs in the city of São Luís - MA, Brazil

Babesiosis is an emerging, worldwide disease caused by protozoa of the genus Babesia, which infects domestic animals and wildlife, as well as humans. The objective of this study was to determine the occurrence of Babesia canis vogeli in dogs in the city of São Luís - MA, Brazil. Blood samples were collected from 65 animals for direct examination, through blood smear and molecular diagnosis, through PCRun. Among the 65 animals analyzed by direct diagnosis, none were positive for Babesia spp. while PCRun showed that 5 (7.69%) were infected by B. canis vogeli, demonstrating the presence of this species in dogs from an urban environment in the city of São Luís. It was concluded that PCR is a more sensitive and specific for this diagnosis, and can be used to define more precisely the infection of dogs by B. canis vogeli. This study confirmed the presence of B. C. vogeli in dogs in the city of.

Standardization of recombinant Ancylostoma caninum cysteine protease 2 (rAcCP2) based indirect ELISA for serodiagnosis of hookworm infection in dogs

Diagnosis of hookworm infection in dogs during pre-patentency or in pregnant bitches harbouring encysted larvae in tissues can be achieved by employing serological tests using proteins derived from somatic or excretorysecretory products of adult or larvae of Ancylostoma caninum. In the present study, cathepsin-B protease (AcCP2) of A. caninum, which helps in development of parasitism and nutrient digestion, was used to develop an indirect ELISA for detection of specific antibodies to A. caninum in dogs. The rAcCP2 (approx. 59.0 kDa) was cloned, expressed and purified under denaturing conditions. Serum samples of 20 known A. caninum positive and 15 known negative dogs were used for the standardization of indirect ELISA. The sensitivity and specificity of the assay was determined by using sera samples from 123 dogs (48 positive for A. caninum eggs in faeces and 75 faecal negative). Out of the 48 A. caninum faecal positive sera, 46 were tested positive (OD &gt; 0.306) by indirect ELISA; whereas, 14 out of 75 faecal negative samples were also tested positive (OD &gt; 0.306) by indirect ELISA. Based on the results, the sensitivity and specificity of rAcCP2 based indirect ELISA was calculated to be 95.8% and 84.3%, respectively. No cross-reactions were observed with serum from dogs naturally infected with B. canis vogeli, B. gibsoni, E. canis, Dirofilaria immitis and Toxocara canis. The results of the present study indicate that indirect ELISA with rAcCP2 protein might be appropriate in large scale epidemiological screening and for serological diagnosis of A. caninum infection in dogs.

First Molecular Detection of Piroplasm Infection in Pet Dogs from Gansu, China.

Babesiosis, the hemolytic disease caused by Babesia, which is a tick-transmitted obligate intraerythrocytic protozoan parasite. This disease is responsible for significant mortality and morbidity rates and enormous economic losses to the livestock industry in tropical and subtropical regions worldwide. In this study, blood samples were collected from 141 pet dogs from Gansu, China, and analyzed for Babesia or Theileria spp. infection using specific PCR and sequencing based on 18S rRNA gene fragments. The results indicated that 18S rRNA gene sequences from 11 samples were similar to the 18S rRNA gene sequences in Babesia canis vogeli (2) and Theileria sinensis (9). The total infected rates of B. canis vogeli and T. sinensis were 1.4% (2/141) and 6.4% (9/141), respectively. This represents the first molecular report of T. sinensis in dogs worldwide and of B. canis vogeli in dogs from Gansu province of China. Furthermore, the finding of T. sinensis in dogs may represent the common infection of this parasite occurring in Gansu.

WITHDRAWN:First report on Babesia canis vogeli infection in dogs in the Philippines.

The Publisher regrets that this article is an accidental duplication of an article that has already been published, http://dx.doi.org/10.1016/j.parint.2016.10.001.The duplicate article has therefore been withdrawn. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.