• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.103-109
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• 2004

Babesia equi ema-1 5' intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5' IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes (Bgll , HindⅢ, Kpn1 and BamH1) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.219-221
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• 1998

This report was conducted to determine the changes of blood and serum chemical values on dogs infected with Babesia gibsoni. Blood and serum were obtained and analyzed with a blood chemical analyzer and calculated by Chauvent's statistical analysis. The mean values and SD were as follows, $AST 33{\pm} 23 IU/L, ALT 22{\pm}15 IU/L, BUN 18$\pm $8 mg/dl, creatinine 1.6{\pm}0.8 mg/dl, CPK 564{\pm}214 IU/L, LDH 208{\pm} 78 IU/L, ALP 52{\pm} 8 IU/L, glucose 89{\pm}38 mg/dl, RBC 3.04{\pm}0.77{\times}10^{6}/{\mu}l, hemoglobin 6.1{\pm} 1.3 g/dl, PCV 18.0{\pm }4.3%, $respectively.

• 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).

• 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.

• Parasites, Hosts and Diseases
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• v.52 no.5
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• pp.507-511
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• 2014

Outbreaks of tick-borne disease cases in Santa Catarina, Brazil are known, but the presence of the pathogen DNA has never been determined. In this study, the first survey of Anaplasma marginale, Babesia bigemina, and Babesia bovis DNA on blood samples of 33 cattle from an outbreak in Ponte Alta Municipality, Santa Catarina, Brazil, has been carried out. A multiplex PCR detected 54.5% of animals were co-infected with 2 or 3 parasites, while 24.2% were infected with only 1 species. The most prevalent agent was B. bigemina (63.6%) followed by A. marginale (60.6%). This is the first report of tick-borne disease pathogens obtained by DNA analysis in Southern Brazil.

• 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.

• 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.

• Korean Journal of Veterinary Service
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• v.27 no.1
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• pp.95-101
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• 2004

Babesia bovis rap-1 and B equi ema-1 intergenic(IG) nucleotides were analyzed and compared for identifying putative promoter sites using computer programs. The reason to initiate this research was to determine if IG nucleotides of Babesia genes that are predicted to be involved in erythrocyte invasion have functions regulating gene transcription and translation, which can be applied to functional gene knockout. Four IG sequences used included BbIG5(B bovis rap-1 5' IG), BblG3(B bovis rap-1 3' IG), BeIG5(B equi ema-1 5' IG) and BeIG3(B equi ema-1 3' IG). BbIG5 contained a putative promoter at nucleotide 197-246 with a predicted TATA-box and a transcription start site. BbIG3 had a putative promoter at nucleotide 270-320 with two predicted TATA-boxes and a transcription start site. BeIG3 had a putative promoter at nucleotide 155-205 with a predicted TATA-box and a transcription start site. Putative promoter sites in these three sequences mentioned above were identified with score cutoff 0.8, which means detection of about 40% recognized promoters with 0.1-0.4% false positives. In contrast, BeIG5 had a putative promoter at nucleotide 163-213 with score cutoff 0.8, but neither TATA-box nor transcription start site were recognized. However, BeIG5 had a putative promoter at nucleotide 388-438 with a predicted TATA-box and a transcription start site when score cutoff was decreased to 0.18, which means detection of about 70% recognized promoters with 2.2-5.3% false positives. These sequences with putative promoters can be tested if they have functions regulating gene transcription and translation.

• 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.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.