Babesia-infected Dogs Research Articles

The Diverse Pathogenicity of Various Babesia Parasite Species That Infect Dogs.

Babesia species infect a very wide range of mammal hosts across the globe, and zoonotic infections are of growing concern. Several species of the Babesia genus infect dogs, and some of these cause significant morbidity and mortality. The Apicomplexan parasite resides within the red cell and infections result in direct damage to the host through intra- and extravascular hemolysis. An exuberant inflammatory response by the host to some species of Babesia parasites also results in significant collateral damage to the host. Canine infections have been the subject of many studies as the well-being of these companion animals is increasingly threatened by the spread of tick vectors and an increasingly mobile dog population. There are currently no widely available and effective vaccines, and effective treatment can be challenging. Understanding disease pathogenesis underlies the development of new treatments. The varying pathogenicity of the various Babesia parasite species that infect dogs offers an opportunity to explore the molecular basis for the wide range of diseases caused by infection with this parasite genus. In this review, we focus on what has been reported about the clinical presentation of Babesia-infected dogs in an attempt to compare the severity of disease caused by different Babesia species.

The First Records of Canine Babesiosis in Dogs from Dermacentor reticulatus-Free Zone in Poland.

Tick-borne microorganisms belong to important etiological agents of many infectious diseases affecting humans and animals. Among them, there are haemoprotozoans of the Babesia genus, which infect erythrocytes of a host and may cause many clinical symptoms. Canine babesiosis is an emerging tick-borne disease in Southern and Central Europe. In this study, we report two cases of symptomatic canine babesiosis caused by Babesia canis in domestic dogs from the Silesian Voivodeship, Poland, as well as the presence of Dermacentor reticulatus ticks detected on one of the Babesia-infected dogs (D. reticulatus-free zone). The molecular analysis confirmed the presence of Babesia canis in the dogs’ blood, and the sequencing analysis showed that the obtained sequence is 100% identical to the sequence of Babesia canis isolate 3469 (sequence ID: KX712122.1). Our findings should raise awareness of B. canis infection among dog owners and veterinarians in the region where B. canis was not previously reported in residential, non-traveling dogs, as well as ensuring that adequate diagnostic methods are available.

Molecular Identification and Genotyping of Babesia canis in Dogs from Meshkin Shahr County, Northwestern Iran.

Background:Canine babesiosis is one of the mainly worldwide-distributed tick-borne haemoprotozoan parasitic diseases in dogs.Methods:A total of 43 blood samples were randomly collected from naturally infected dogs in seven villages from different geographical areas of Meshkin Shahr, Ardabil Province, Iran. The presence of Babesia species detected with standard methods including parasitological and gene sequencing techniques targeting the 18S rRNA gene.Results:Our results revealed that four dogs 9.3% (4/43) including one female and three male dogs were infected with Babesia. All four Babesia-infected dogs were confirmed B. canis by the molecular-based method. Sequence alignments comparison of the B. canis genotypes A and B, it was revealed that all B. canis isolates belonged to genotype B.Conclusion:This study provides essential data for subsequently define the critical importance of the molecular studies in management and prevention of the canine babesiosis in Iran.

Serum Hepcidin as a Clinical Prognostic Marker to Discriminate the Outcome of Canine Babesiosis

Background: We aimed to identify the prognostic value of hepcidin in discriminating against the survival outcome of canine babesiosis. Methods: Semi-nested polymerase chain reaction was performed to confirm the presence of infection. Existence of oxidative stress and inflammatory response, changes in systemic iron status and hepcidin level were assessed in the study population. Based on the outcome Babesia infected dogs were classified into survivors (n=18) and non-survivors (n=14) of infection. 32 healthy dogs formed the control group. Conclusion: In non-survivors of infection, serum hepcidin was positively associated with C-reactive protein (P less than 0.01), serum iron (P less than 0.01), transferrin iron-binding capacity (P less than 0.05), unsaturated iron-binding capacity (P less than 0.01), thiobarbituric acid reactive substance (P less than 0.05) and negatively associated with catalase (P less than 0.01), zinc (P less than 0.01) and low haemoglobin density (P less than 0.01). The prognostic cut-off value of hepcidin in discriminating the survivability of infected dogs was 32.32 ng/mL with 100.00% specificity and 92.86 % sensitivity. The area under the curve of hepcidin in discriminating survivability was about 0.984 and Youden’s index was 0.928. Hence, hepcidin can predict the survival outcome of the disease enabling intensive care for animals with a cut-off value of hepcidin more than 32.32 ng/mL.

Evaluation of acute kidney injury in dogs with complicated or uncomplicated Babesia rossi infection

Dogs with babesiosis can present with multiple complications, including acute kidney injury (AKI). The objective of this study was to characterize AKI in dogs with babesiosis caused by Babesia rossi at presentation and after treatment. Thirty-five client-owned dogs with B. rossi infection and 10 control dogs were included in this prospective observational study. Blood and urine were collected in Babesia-infected dogs at presentation (T0, n = 35), after 24 h (T24h, n = 11), and after 1 month (T1m, n = 9). The following urinary kidney injury biomarkers were assessed: urinary protein to creatinine ratio (UPC), urinary glomerular injury biomarkers (immunoglobulin G (uIgG) and C-reactive protein (uCRP)), and urinary tubular injury biomarkers (retinol-binding protein (uRBP) and neutrophil gelatinase-associated lipocalin (uNGAL)). Serum functional renal biomarkers were creatinine (sCr) and symmetric dimethylarginine (sSDMA). Post-mortem kidney biopsies were analyzed by light and transmission electron microscopy.At T0, all kidney injury biomarkers were significantly higher in Babesia-infected dogs compared to healthy controls (P < 0.001), while functional renal biomarkers were not significantly different (P > 0.05). At T24h, all urinary tubular injury biomarkers and UPC decreased significantly (P < 0.01), while glomerular injury biomarkers did not (P = 0.084). At T1m, all urinary kidney injury biomarkers decreased to values not significantly different from healthy controls (P > 0.5). Significant changes in functional renal biomarkers were not seen after treatment (P > 0.05). Dogs with complicated babesiosis had significantly higher glomerular injury biomarkers, UPC, and sSDMA compared to uncomplicated cases (P < 0.05), while all tubular injury biomarkers and sCr were not significantly different (P > 0.1).Dogs with babesiosis caused by B. rossi showed transient kidney injury, which was detected by all kidney injury biomarkers, but remained undetected by functional biomarkers. All infected dogs, irrespective of disease severity, suffered comparable kidney injury based on tubular injury biomarker concentrations, while loss of function was seen more often in dogs with complicated babesiosis based on sSDMA results.

Neutrophil Myeloperoxidase Index in Dogs With Babesiosis Caused by Babesia rossi.

Babesiosis caused by the virulent tick-borne hemoprotozoan, Babesia rossi, results in a marked systemic inflammatory host response in dogs. Neutrophils form part of the innate immune response and contains myeloperoxidase (MPO) as the predominant component of the neutrophil lysosomal protein in azurophilic granules. The neutrophil myeloperoxidase index (MPXI), determined on the ADVIA hematology analyzer, is a quantitative estimate of intracellular MPO content. Objectives of this study were to: (a) compare MPXI in dogs with babesiosis with healthy control dogs; (b) compare MPXI in dogs that died from babesiosis with dogs that survived and controls; and (c) correlate the MPXI with the previously determined segmented and band neutrophil count and cytokine concentrations in dogs with babesiosis. Data for 140 dogs naturally infected with B. rossi and 20 healthy control dogs were retrospectively evaluated. Neutrophil counts and MPXI were determined on an ADVIA 2120 analyzer. Cytokine concentrations [interleukin (IL)-2, IL-6, IL-8, IL-10, IL-18, granulocyte-macrophage colony stimulating factor (GM-CSF), and monocyte chemo-attractant protein-1 (MCP-1)] were determined using a canine-specific multiplex immunoassay. The mortality rate of the Babesia-infected dogs was 11% (15/140). MPXI was significantly higher in Babesia-infected dogs (P = 0.033), and in Babesia-infected non-survivors (P = 0.011), compared with healthy control dogs. In Babesia-infected dogs a significant positive correlation was found between MPXI and IL-10 (r = 0.211, P = 0.039) and a significant negative correlation was found between MPXI and IL-8 (r = −0.350, P < 0.001). In Babesia-infected non-survivors, significant positive correlations were found between MPXI and IL-2 (r = 0.616, P = 0.033), IL-6 (r = 0.615, P = 0.033), IL-18 (r = 0.613, P = 0.034), GM-CSF (r = 0.630, P = 0.028), and MCP-1 (r = 0.713, P = 0.009). In Babesia-infected survivors, a significant negative correlation was found between MPXI and IL-8 (r = −0.363, P = 0.001). MPXI was correlated with pro-inflammatory cytokines in Babesia-infected dogs that died. The potential of MPXI as a novel marker of inflammation and prognosis in dogs infected with B. rossi, thus warrants further investigation.

Molecular diagnosis and clinico-hemato-biochemical alterations and oxidant-antioxidant biomarkers in Babesia-infected dogs of Mizoram, India.

Babesiosis is a tick transmitted disease, infecting a wide variety of wild and domestic animals, as well as humans. This study was designed to investigate molecular diagnosis and clinic-hemato-biochemical and oxidant/antioxidant status in dogs of Mizoram, India. A total 1200 dogs screened for babesiosis during 2017-18 and 53 dogs suspected for babesiosis by clinical signs and were confirmed by molecular diagnosis. Clinical signs were recorded; also blood samples were taken to investigate hematologic changes, serum biochemical variations and oxidative stress biomarkers. The overall incidence of babesiosis in dogs of Aizawl, Mizoram, India during the study period recorded was 1.25% (15/1200) and 28.3% cases confirmed from 53 suspected dogs (15/53). The most commonly observed clinical signs were fever, emaciation, depression and icterus and lymphadenopathy. Significant reduction in PCV, HB, RBCs, MCHC, total protein, and albumin along with significant increase in MCV, WBCs, monocytes and BUN were the most consistent hemato-biochemical changes. Oxidant/antioxidant assessment showed significant reduction in superoxide dismutase, catalase and total anti-oxidant (TAC) along with significant increase in lipid peroxidase (LPO) activities. The findings of this study demonstrated that the main causative agent of babesiosis in dogs in Mizoram Province is Babesia gibsoni which caused significant alteration of hemato-biochemical and oxidant-antioxidant status in dogs.

A flow cytometric assessment of the lymphocyte immunophenotypes in dogs naturally infected with Babesia rossi

Immunity to Babesia infection requires both innate and acquired responses, including cell mediated- and humoral responses. The aims of this study were to investigate the variation in selected peripheral blood lymphocyte phenotypes in dogs with virulent babesiosis at presentation and over time after treatment, and to determine whether these were correlated with the severity of clinical signs. Forty-four dogs naturally infected with B. rossi were studied and 5 healthy dogs were included as controls. Blood samples were collected from the jugular vein at admission, prior to any treatment, and at 24h and 48–72h. Leukocytes were incubated with canine specific, fluorochrome conjugated anti-CD3, anti-CD4, anti-CD8, and anti-B cell markers. Babesia-infected dogs were divided into complicated or uncomplicated groups on clinical grounds and in-house laboratory assays. The percentage CD3+ lymphocytes in the complicated group was lower compared to the controls (P=0.014) and uncomplicated group (P=0.007). The percentage CD4+ T lymphocytes in the complicated group was lower compared to the controls (P=0.027) and uncomplicated group (P=0.014). Both the complicated as well as the uncomplicated groups expressed a lower percentage CD8+ T lymphocytes compared to the control group (P<0.001 and P=0.005, respectively). The percentage B lymphocytes was higher in the complicated group at 48–72h. These findings could indicate the presence of a functional immune suppression secondary to increased apoptosis or redistribution of effector lymphocytes and/or a combination of other immune modulatory mechanisms induced by B. rossi infection.

Excessive Pro-Inflammatory Serum Cytokine Concentrations in Virulent Canine Babesiosis

Babesia rossi infection causes a severe inflammatory response in the dog, which is the result of the balance between pro- and anti-inflammatory cytokine secretion. The aim of this study was to determine whether changes in cytokine concentrations were present in dogs with babesiosis and whether it was associated with disease outcome. Ninety-seven dogs naturally infected with B. rossi were studied and fifteen healthy dogs were included as controls. Diagnosis of babesiosis was confirmed by polymerase chain reaction and reverse line blot. Blood samples were collected from the jugular vein at admission, prior to any treatment. Cytokine concentrations were assessed using a canine-specific multiplex assay on an automated analyser. Serum concentrations of interleukin (IL)-2, IL-6, IL-8, IL-10, IL-18, granulocyte-macrophage colony stimulating factor (GM-CSF) and monocyte chemotactic protein-1 (MCP-1) were measured. Twelve of the Babesia-infected dogs died (12%) and 85 survived (88%). Babesia-infected dogs were also divided into those that presented within 48 hours from displaying clinical signs, and those that presented more than 48 hours after displaying clinical signs. Cytokine concentrations were compared between the different groups using the Mann-Whitney U test. IL-10 and MCP-1 concentrations were significantly elevated for the Babesia-infected dogs compared to the healthy controls. In contrast, the IL-8 concentration was significantly decreased in the Babesia-infected dogs compared to the controls. Concentrations of IL-6 and MCP-1 were significantly increased in the non-survivors compared to the survivors. Concentrations for IL-2, IL-6, IL-18 and GM-CSF were significantly higher in those cases that presented during the more acute stage of the disease. These findings suggest that a mixed cytokine response is present in dogs with babesiosis caused by B. rossi, and that an excessive pro-inflammatory response may result in a poor outcome.

Platelet indices in dogs with Babesia rossi infection.

Thrombocytopenia without clinical bleeding is a consistent finding in virulent canine babesiosis. The purpose of the study was to investigate the platelet index phenotype in Babesia rossi-infected dogs and the association with disease outcome. We hypothesized that an increased proportion of large, activated platelets would be present. Ninety-six infected and 15 control dogs were included. Babesia-infected dogs were further divided into survivors and nonsurvivors. Platelet count, mean platelet volume (MPV), platelet volume distribution width (PDW), plateletcrit (PCT), mean platelet mass (MPM), mean platelet component concentration (MPC), and platelet component distribution width (PCDW) were measured at presentation, and at 24 and 48 hours in admitted survivors. Mortality rate was 13% (12/96). At presentation, compared to controls, PLT and PCT were significantly decreased in survivors and nonsurvivors (P < .001 for both). Mean platelet volume was significantly increased in survivors and nonsurvivors compared to the controls (P < .001 for both); however, MPM was only significantly increased in the survivors (P < .001). There were no differences between the survivors and nonsurvivors for any of the indices at presentation. Platelet count and PCT were significantly increased at 24 and 48 hours, and MPC significantly increased at 24 hours for admitted survivors compared to the values at presentation. Large, activated platelets were significantly increased and may play a role in the lack of a bleeding phenotype, despite severe thrombocytopenia, in canine babesiosis.

Platelet activation and platelet–leukocyte interaction in dogs naturally infected with Babesia rossi

Using flow cytometry, platelet–leukocyte aggregate (PLA) formation has previously been documented in dogs with a variety of systemic inflammatory disorders and immune-mediated haemolytic anaemia. Platelet activation and subsequent interaction between platelets and leukocytes are important for regulating innate immunity and systemic inflammation. The objective of this study was to investigate PLA formation in canine babesiosis and to determine whether it was associated with outcome. Blood was collected from 36 client-owned dogs diagnosed with Babesia rossi infection and 15 healthy controls using EDTA as anticoagulant. Activated platelets and PLA formation were detected by measuring surface expression of P-selectin (CD62P) on platelets, monocytes and neutrophils.Of the Babesia-infected dogs, 29 survived and seven died. The percentage of CD62P-positive monocytes was significantly higher (P = 0.036) in the Babesia-infected dogs (54%) than in healthy control dogs (35.3%). However, there were no significant differences between the Babesia-infected and control groups for CD62P-positive platelets (4.9% and 1.2%, respectively) and CD62P-positive neutrophils (28.3% and 17.9%, respectively). The percentage of CD62P-positive monocytes was significantly higher (P = 0.019) in the survivors (58.9%) than in healthy control dogs; however, there were no significant differences between the non-survivors (39.2%) and the controls or between survivors and non-survivors. There were no significant differences between groups for the percentage of CD62P-positive platelets (survivors 4.8%; non-survivors 5.3%; controls 1.2%) or CD62P-positive neutrophils (survivors 31.6%; non-survivors 5.6%; controls 17.9%). In conclusion, Babesia-infected dogs, specifically dogs that survived, had a significantly increased percentage of platelet–monocyte aggregates compared to healthy control dogs.

Clinical, Hematologic, and Molecular Findings in Naturally OccurringBabesia canis vogeliin Egyptian Dogs

Background. Canine babesiosis is a clinically important hemoprotozoan parasite affecting dogs. The goal of this present study was to determine the clinical symptoms and to establish its hematological and microscopic detection and compare it with the PCR findings attained from dogs infected with Babesia canis vogeli. Methodology/Principal Findings. 13-PCR confirmed Babesia-infected dogs were examined; seminested PCR was used to discover the precise type of Babesia and Babesia canis vogeli was the only subspecies detected. The most consistent clinical signs were elevated rectal temperature and a pale mucous membrane. Thrombocytopenia, monocytosis, and lymphocytosis, along with a significant reduction in red cell parameters, were the most commonly recorded hematologic alterations. Microscopic examination revealed the presence of typical large merozoites and trophozoites of B. canis in the ratio 76.92%. Conclusions/Significance. The presumptive diagnosis of canine babesiosis should be based on a fever and anemia, while thrombocytopenia is considered the hallmark of the disease; microscopic examination may not be very revealing in the detection at low parasitemia, but it remains the most rapid confirmatory method. Seminested PCR turned out to be a sensitive and accurate method for diagnosis; during the process of differentiation between Babesia subspecies, only B. canis subsp. vogeli was detected.

The effect of pyridoxal-5-phosphate on serum alanine aminotransferase activity in dogs suffering from canine babesiosis

Accurate measurements of serum aminotransferase (ALT) activity in dogs relies on the endogenous pro-enzyme pyridoxal 5-phosphate (P5P). The purpose of this study was to determine whether the exclusion of P5P from the analytical method causes an underestimation of serum ALT activity in dogs suffering from babesiosis and in those manifesting evidence of hepatocellular damage, and to determine if anorexia causes sufficient P5P depletion to affect in vitro serum ALT activity. One-hundred-and-twenty healthy control dogs and 105 Babesia-infected dogs were included in the study. Two methods for ALT measurement were used: Method 1 included P5P, and Method 2 excluded P5P from the reaction mixture. Higher serum ALT activity was measured with Method 1 in the Babesia-infected dogs (P < 0.001), as well as in 14 dogs with suspected hepatocellular damage (P = 0.03). Duration of anorexia had no effect, irrespective of the method used. Although inclusion of P5P to the reaction mixture consistently resulted in higher measured serum ALT activity, the differences were too small to have led to incorrect diagnoses in the Babesia-infected dogs suspected of liver disease.

Clinical and pathological findings of Babesia infection in dogs

The clinical and pathological findings of Babesia infection in 32 dogs in northern Australia are presented. Eleven different breed types were represented from 6 localities in north Queensland and one locality in northern Western Australia. Twenty three (72%) were males. Babesia-infected dogs were grouped by the degree of haematological disturbance and clinical severity: Acute babesiosis (25/32), all pups with severe haemolytic anaemia; subclinical carriers (5/32) with non-specific malaise, characterised haematologically by a normal erythrogram but marked leucopenia; chronic anaemia, observed in 2 adult dogs. Pups were azotaemic (serum urea greater than 6.6 mmol/l) and had elevated serum bilirubin levels (20.8 to 48.5 mmol/l). Total serum protein was usually within the normal range. Pups that died were also hypoglycaemic and severely hyperkalaemic (K+ greater than 10 mmol/l). Low parasitaemias in routine blood smears complicated diagnosis but smears made from ear or toe capillaries, or after haematocrit concentration, greatly enhanced finding parasitised cells. At necropsy, pallor and jaundice were the most consistent observations. Haemoglobinuric nephrosis, an active reticulo-endothelial system and capillaries packed with large numbers of infected erythrocytes were the main histopathological findings. A combination of imidocarb dipropionate at 5 mg/kg body weight, given intramuscularly, with fluid therapy and blood transfusion was the most successful treatment.