• 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.

• Proceedings of the Korean Society of Veterinary Clinics Conference
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• 2008.10a
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• pp.154-154
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• 2008

• 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.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.89-97
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• 1991

For the ultrastructural observation on Babesia gibsoni(B gibsoni), the protozoa were challenged experimentally to splectomized dog. To examine the ultrastructure of the B gibsoni in the erythrocyte, the infected erythrocytes were collected at the cephalic or jugular vein of the dog. The results obtained by TEM(transmission electron microscopy) were as follows; 1. The sizes of protozoa in erythrocytes are $0.92{\pm}0.36{\mu}m{\times}0.67{\pm}0.21{\mu}m$, the sizes of nucleus of the protozoa are $0.55{\pm}0.24{\mu}m{\times}0.38{\pm}0.26{\mu}m$, and sizes of rhoptries in plasma of the protozoa are $0.33{\pm}0.05{\mu}m{\times}0.25{\pm}0.07{\mu}m$, respectively. 2. The tropozoite membrane in the erythrocyte was one, and it's nuclear membrane was made up of double. But the protozoa of initial stage in infected erythrocyte had double clear mambranes, and distinguished from plasma membrane of red blood cell. 3. The mitochondrialike structures covered with two membranes were observed in the protozoa. 4. Mitochondria and vesicles of the reticulocyte were observed near protozoa in the erythrocyte. 5. There are rhoptry, coiled structure and single nucleous in the merozoite. 6. The shape of rhoptry was round or ovoid form and in occasionally, the content of rhoptry was lost partially. 7. There was able to observe the dividing process of the protozoa. 8. Maurer's cleft-like structure was observed.

• Journal of Veterinary Clinics
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• v.37 no.1
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• pp.23-27
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• 2020

Between May and November 2018, babesiosis was examined in 162 bloods samples obtained to an animal hospital in Jeju island for anemia and medical examination. Sixty-two of 162 (38.3%) were positive by PCR. The ultra fast real-time PCR test with blood directly analyzed without DNA extraction showed the same results. Accurate diagnosis, treatment and prognosis of babesiosis should be combined with clinical symptoms, blood tests, the babesia antibody test, and the PCR antigen test. Ultra fast real-time PCR, with these tests, is expected to be a point-of-care testing (POCT) for easy, fast and accurate diagnosis of babesiosis in the veterinary clinic.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

• Journal of Veterinary Clinics
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• v.39 no.5
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• pp.207-216
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• 2022

Canine babesiosis has been scarcely investigated in the Republic of Korea (ROK). Although it is known that Babesia gibsoni is its primary causative agent, its clinical presentation has not been completely clarified in the ROK. Consequently, the aim of this study was to evaluate the clinical appearance of this parasitic infection based on the anamnesis of the patient and compare of hematological and biochemical test results. Four hundred whole blood samples from patients with a presumptive diagnosis of tick-borne disease were analyzed by polymerase chain reaction (PCR) to amplify the Babesia spp. 18S rRNA gene and by a rapid diagnostic test kit (VetAll Laboratories^®) to detect B. gibsoni seroreactive animals. Thirty-six (9.0%) dogs were PCR-positive but only 24 (6.0%) were seropositive. The investigation revealed that all the courses of the disease are present in the ROK, with the acute course being predominant. The acute course tends to consist of inappetence, lethargy, pyrexia, gastrointestinal symptoms, and occasionally hematuria. It also occurs with common hematological abnormalities, such as thrombocytopenia and anemia, and to a lesser extent biochemical abnormalities, such as hyperbilirubinemia, hypoalbuminemia, and elevated liver enzymes. This research shows that B. gibsoni is an endemic hemoparasite capable of producing a variety of clinical manifestations in dogs. For its accurate diagnosis, a descriptive history of the clinical signs, hematology, and biochemical profile of the patient, along with a well-performing PCR assay, are essential. These findings will help in planning pragmatic preventive strategies against this potent threat in the ROK.

• Korean Journal of Veterinary Research
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• v.46 no.1
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• pp.77-81
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• 2006

A 1-year and 6-month-old, intact male, Shih-tzu dog was referred to the Konkuk University Veterinary Teaching Hospital, due to ascite, melena, severe anemia, and polyuria/polydipsia. Complete blood count showed moderate leukocytosis, lymphocytosis, monocytosis, and microcytic hypochromic nonregenerative anemia. On serum biochemistry profiles, hypoalbuminemia and hypoproteinemia were observed, and proteinuria was detected on urinalysis. Pleural effusion and mild ascites were noted in radiological findings, and bilateral nephromegaly was noted on ultrasonography. Babesia gibsoni was detected by PCR. Babesiosis and glomerular disease was treated at a time with prednisolone and buparvaquone. This therapeutic regime was very efficacious and clinical sings were rapidly improved. Proteinuria and severe anemia were disappeared following therapy.

• Journal of Veterinary Clinics
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• v.6 no.1
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• pp.185-191
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• 1989

A dog which was hospitalized to Veterinary Teaching Hospital, College of Veterinary Medicine, Jeonbug National University on December 28, 1988 was revealed severe anemia: hemoglobinuria and weakness. In the inspections, abdominal pain and spleno megaly at the ventral abdomen were detected by palpations. In the examinations of blood, the obtained results were summarized as follows: Babesla spp. was identified on the blood smear stained with Giemsa. The Babesia spp. was assumed to the Babesia gibsoni for the their small size and pleomorphism such as comma form, ring form and dot form. In the blood examinations of the patient, Ht: 22.5％, RBC:354${\times}$10$^4$/${\mu}\ell$, Hb: 8.8g/dl, serum protein: 8g/dl, and WBC count was 21, 425/${\mu}\ell$. In the chemical examinations of serum, the value of AST(GOT) was 30iu and ALT(GPT) was 20iu, respectively. The blood sugar was 60mg/d1. In the urine test, urine protein was 30mg/d1 and the hemoglobin In the urine was the ＋＋＋ and occult blood reaction(Benzidine test) in the feces was ＋＋＋. Splenomegaly was confirmed by X-ray examination. To confirm for the Babesia spp. infection, 5ml of the whole blood of the patient(3％ of Parasitized erythrocytes) were inoculated into the cephalic vein of the two normal dogs. In the blood of experimental dogs which were inoculated parasitized blood, Babesia spp. was detected in the two doss and pleomorphic parasites were observed, too. In the blood examinations of No. 1 the Ht and RBC were decreased to 6.8％ and 52${\times}$10$^4$/${\mu}\ell$, respectively. WBC count was 10.600/${\mu}\ell$ and serum protein was 6.8g/dl. The rates of parasitized erythrocytes were 15％ in the experimental dog. Also ＋＋＋ of the hemoglobin was detected in the urine. In the X-ray examination, splenomegaly was comfirmed and it was confirmed by autopsy of the experimental dog(No. 1).