• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.359-365
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• 1998

This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

• Journal of Veterinary Clinics
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• v.33 no.1
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• pp.16-20
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• 2016

This study was performed to estimate the clinical use of combination therapy with atovaquone/proguanil and azithromycin as a effective treatment in dogs infected with Babesia gibsoni. Eight mixed-breed dogs that were infected naturally with B. gibsoni were used in this study. Four dogs (No. 1-4) as experimental group received atovaquone/proguanil and azithromycin therapy. As for the other four dogs as the control group (No. 5-8) were administered diminazene aceturate and tetracycline/clindamycin. All the dogs in this study showed mild to severe anemia and thrombocytopenia. After initiating the treatment B. gibsoni in blood smears disappeared. PCR analysis of the experimental group showed negative results during the observation period, but more than one dog from the control groups showed continuous positive results. Atovaquone/proguanil and azithromycin combination therapy can significantly lower the B. gibsoni parasitemia levels and the results suggested that this combination therapy should be a new protocol for an effective treatment in dogs infected with B. gibsoni.

• Journal of Veterinary Clinics
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• v.4 no.2
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• pp.505-514
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• 1987

Babesia gibsoni infection was diagnosed and treated in three hunting dogs which were hospitalized to the Veterinary Teaching Hospital, College of Veterinary Medicine, Seoul National University between April 4 and August 29, 1987. All three dogs revealed severe anemia, hemoglobinuria, splenomegaly and markedly decreased PCV, RBC count and hemoglobin. The anemia was regenerative, as characterized by increased numbers of nucleated erythrocytes, polychromasia, anisocytosis, reticulocytosis. B. gibsoni was identified by examination of blood smear stained with Giemsa stain. The forms of B. gibsoni identified in this report were pleomorphic such as singnet ring, oval, comma, dot and elongated forms. The maximal percentages of erythrocytes infected with one or more B. gibsoni organisms were 39%, 20% and 40%, respectively. The Tick, Haemophysalis longicornis was assumed to be the vector of babesiosis in these cases. Specific treatment consisted of diminazene aceturate and supportive treatment consisted of whole blood transfusion, lactated Ringer's solution, vitamin B complex and broad spectrum antibiotics. All three dogs had convalesced successfully after treatment.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.101-108
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• 1993

To examine the effects of vaccination against Babesia gibsoni(B gibsoni) infection in dogs, 15 normal mixed-breed dogs(5 months to 1 year old) were divided into 3 groups with 5 dogs in each group. One of them was selected as control group(group A) and other were selected as experimental groups(group B and C). The group B was vaccinated with antigen which were mixed 0.2% of formalin treated B gibsoni and sonicated one. The group C was inoculated Theileria sergenti as a non-specific antigen. The results obtained in this experiment were summarized as follows; 1. After first vaccination, antibody titers of group B and C were increased 5 times(1:200) than those of control group(1 : 40). The antibody titers of group C were increased more than that of group B after second vaccination. When challenged with the living protozoa(Babesia gibsoni), the antibody titers of C group were elevated higher than that of B group and maintained steadly. Those were not exceeded over 1 : 5,000 for 4 weeks in all 3 groups. 2. After challenge, the peak time of the parasitemia appeared nearly on the 15th day(12~18 days) in all groups. During this period, the rate of parasitized erythrocytes in control group was $55.0{\pm}5.4$‰. But that of group B and C were $41.3{\pm}38.8$‰ and $15.2{\pm}16.3$‰, respectively. 3. After challenge with B gibsoni, all of the values of PCV, Hb, RBC and total leukocytes counts were decreased in both of the experimental and the control. 4. In all groups, there were increased lymphocytes and monocytes after challenge with the protozoa.

• Parasites, Hosts and Diseases
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• v.52 no.4
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• pp.345-353
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• 2014

Babesia gibsoni is an intraerythrocytic apicomplexan parasite that causes piroplasmosis in dogs. B. gibsoni infection is characterized clinically by fever, regenerative anemia, splenomegaly, and sometimes death. Since no vaccine is available, rapid and accurate diagnosis and prompt treatment of infected animals are required to control this disease. Over the past decade, several candidate molecules have been identified using biomolecular techniques in the authors' laboratory for the development of a serodiagnostic method, vaccine, and drug for B. gibsoni. This review article describes newly identified candidate molecules and their applications for diagnosis, vaccine production, and drug development of B. gibsoni.

• Journal of Veterinary Clinics
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• v.34 no.3
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• pp.185-188
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• 2017

This study assessed the seroprevalence of Babesia gibsoni in companion dogs in Korea by enzyme linked immunosorbent assay using recombinant BgTRAP antigen. Dogs were randomly selected from those admitted for various reasons to local private veterinary hospitals and the Animal Medical Center of Chonbuk National University. With the owners' permission, extra blood was drawn from each dog for serological assays. Of the 188 selected dogs, seven (3.72%) were positive for B. gibsoni, including six of 167 (3.59%) indoor and one of 12 (8.33%) outdoor dogs. Of the seven dogs positive for B. gibsoni, four were aged > 10 years, two were < 1 year, and one was between 1 and 10 years; and two were Yorkshires and one each was Shih-tzu, Maltese, Pekinese, beagle and mixed. Concurrent diseases or chief complaints were anemia in two dogs, both of which had a history of confirmed babesiosis by polymerase chain reaction, and non-anemic diseases in five. Geographically, four dogs were from Jeonbuk/Jeonju, and one each from Seoul, Gyounggi-do, and Jeonnam/Gwangju. To our knowledge, this is the first report of companion dogs in Korea being seropositive for B. gibsoni. Serologic screening of subclinical or carrier dogs can detect this potentially dangerous disease and assess its epidemiology.

• Korean Journal of Veterinary Research
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• v.40 no.3
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• pp.587-599
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• 2000

This study was conducted to observe the severity of the disease and pathogenecity of Babesia gibsoni parasite on the splenectomized dogs(SPD) and nonsplenectomized(intact) dogs (NSPD) experimentally infected with B gibsoni. The average prepatent period was 4 days in the SPD and 8 days in the NSPD, respectively. Peak parasitaemia(PE) ranged from 26% to 34% of erythrocytes infected in the SPD and from 4% to 5% in the NSPD. Latent parasitaemia was still detectable 40 days as low as under 1.0% of erythrocytes infected after the initial parasitaemia in the SPD. Blood packed cell volume(PCV) decreased to as little as 6.4% to 6.9% in the SPD. The clinical signs were mild fever and anemia in the NSPD, remissions and exacervations of temperature, intermittent or spike-like increases of temperature, progressive polychromatophilic macrocytic anemia with anisocytosis, icterus, marked loss of appetite, rarely haemoglobinuria, and deep brown-yellowish urine in the SPD. Gross pathologic changes mainly involved slightly enlargement of liver and spleen in the NSPD and marked enlargement of liver in the SPD. Anatomic changes associated with the disease included diffuse periportal and centrilobular hephatitis, and membranoproliferative glomerulonephritis. Hyaline droplets, resulting protein metabolic alterations, were found in the convoluted ephithelium of the kidney. The density of lymphocytes within the liver sinusoids was markedly increased. Aggregates of large monocytes and macrophages were demonstrated in the centrilobular veins of the liver. The density of these cells in the centrilobular veins were greatest in the SPD. The forms of B gibsoni parasite found in the acute stage of SPD were large signet ring form, small signet ring form, pyriform, elongated form, comma form, head-phone form, oval form, peared form, racket-like form, amoeboid form, triangle form, quartered form, dot form, band form and multiple, and rosette form, et al. The severity of the disease and pathogenecity of B gibsoni parasite were mild in the NSPD but fatal in the SPD.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.99-108
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• 1991

To observe the changes of serum proteins according to the process of Babesia gibsoni(B gibsoni) infection, the babesia protozoa($10^8/kg$) were inoculated into the cephalic vein of healthy dogs. The serum proteins of experimentally infected dogs were separated by using cellulose acetate electrophoresis. The results obtained were as follows; 1. Cellulose acetate electrophoresis was fractionated to total 6 of bands such as, albumin, ${\alpha}_1$, ${\alpha}_2$, ${\beta}_1$, ${\beta}_2$ and $\gamma$-globulin. 2. The concentration of total protein was shown a decreasing tendency after B gibsoni infection. Albumin and A/G ratio were lowered through all periods of the infection, but they were not significant changes. 3. The level of ${\alpha}_1$-globulin was significantly(p<0.05) incresed in early stage of the infection. 4. The levels of ${\alpha}_2$ and total $\alpha$-globulin were shown highly significant decreases (p<0.01) through all periods of the infection. 5. The levels of ${\beta}_1$ and total $\beta$-globulin had highly significant changes (p<0.01) that was increased in early stage of infection and decreased later. 6. The level of $\gamma$-globulin was seen to be constantly increased through all periods of infection. It was a highly significant change (p<0.01). 7. Plasma protein: fibrinogen (PP:F) ratio was shown a temporally significant increase (p<0.05) following the decrease in early infection.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.583-593
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• 1997

Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

• Journal of Veterinary Clinics
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• v.7 no.1
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• pp.391-402
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• 1990

To examine the effects of vaccination against Babesia gibsoni infection in dogs, 15 normal mixed-breed dogs(5 month to 1 year old) divided into 3 groups with 5 dogs in a group. One of them was selected as control group(group A) and others were selected as experimental groups(group B and C). The group B was vaccinated with sonicated antigens and the group C was vaccinated with 0.2% of formalin treated antigens. The results obtained in the examination were summarized as follows : 1. In the western blot, the lane A revealed specific two bands on the regions of 54kd and 100kd, respectively. 2. After the first vaccination, the antibody titers of group B and C were higher 5 times(1 ： 200) than those of control group(1：40). After the second vaccination, the antibody titers of group B and C have not changed. When challenged with the protozoa(Babesia gibsoni), the antibody titers(1 : 5,000) were elevated in all groups. But these were not exceeded over 1 ： 5,000 for 4 weeks. 3. After challenge, the peak time of increased numbers of the protozoa was the 15th day (12-18 days) in all groups. During these days, the rate of parasitized erythrocytes in control group was 55.0${\pm}$5.4%. But those of group B and group C were 26.0${\pm}$6.4%, and 15.6${\pm}$7.8％, respectively. 4. After challenge, all of the values of PCV, Hb, RBC were shown to decrease in all of the control and experimental groups. 5. The total leukocytes counts are shown a tendency of reduction in all groups after challenge. 6. In all groups, there were increase in lymphocytes and monocytes after challenge.