Papers by Sivapong Sungpradit
Parasitology research, Jan 27, 2016
Babesia spp., Theileria orientalis, and Anaplasma marginale are significant tick-borne pathogens ... more Babesia spp., Theileria orientalis, and Anaplasma marginale are significant tick-borne pathogens that affect the health and productivity of cattle in tropical and subtropical areas. In this study, we used PCR to detect the presence of Babesia bovis, Babesia bigemina, and T. orientalis in 279 beef cattle from Western Thailand and A. marginale in 608 beef cattle from the north, northeastern, and western regions. The PCRs were performed using species-specific primers based on the B. bovis spherical body protein 2 (BboSBP2), B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. orientalis major piroplasm surface protein (ToMPSP), and A. marginale major surface protein 4 (AmMSP4) genes. To determine the genetic diversity of the above parasites, amplicons of B. bovis and B. bigemina ITS1-5.8s rRNA gene-ITS2 regions (B. bovis ITS, B. bigemina ITS), ToMPSP, and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the prevalence of B. bovis, B. bigemina, T. or...
Veterinary World, Jul 9, 2020
Background and Aim: Canine babesiosis, a tick-borne parasitic disease, is caused by the hemoproto... more Background and Aim: Canine babesiosis, a tick-borne parasitic disease, is caused by the hemoprotozoa, Babesia vogeli, and Babesia gibsoni. Infection with these parasites, which is endemic globally, leads to life-threatening immunosuppression in dogs. The merozoites invade the red blood cells (RBCs) of infected dogs. Ehrlichia canis, an intracellular bacterium that infects monocytes, is transmitted by the same tick species (Rhipicephalus sanguineus) during blood consumption and coinfection with B. vogeli and E. canis has been reported. Although the hematology and biochemistry of canine babesiosis have been studied, more studies are needed to develop a better understanding of the hematobiochemical and molecular profiles associated with cases of single infection and coinfection of canine babesiosis in Thailand. This study aimed to investigate the hematological, biochemical, and molecular profiles of B. vogeli infection and E. canis coinfection.
Materials and Methods: The study included 33 B. vogeli–positive blood samples and 11 E. canis–coinfected blood samples. To exclude coinfection with Hepatozoon canis and Anaplasma platys, only dogs with B. vogeli infection and B. vogeli–E. canis coinfection were included in the study. A multiplex polymerase chain reaction (PCR) assay was conducted to detect B. vogeli, E. canis, and H. canis, and a conventional PCR assay was conducted for the detection of A. platys. Besides, the PCR assay and sequencing, comprehensive data analysis was conducted, including a microscopic blood parasite examination and hematological and biochemical data analysis.
Results: The comparison of the hematobiochemical data between the B. vogeli–positive and E. canis coinfection groups identified that there were statistically significant differences in the RBC parameters, including RBC count, hemoglobin concentration, hematocrit, and RBC distribution width (p=0.001). Neither B. vogeli infection nor coinfection with E. canis was associated with the sex, breed, recorded clinical signs, geographic origin of the dog and also B. vogeli 18S rRNA gene sequencing results.
Conclusion: Coinfection with E. canis increased the severity of babesiosis. The pathogenic mechanisms underlying this infection, such as destruction of RBCs, require further investigation. This study may enhance diagnosis, treatment, and prevention of canine babesiosis.
vogeli, is a health problem in companion animals. The disease causes febrile illness, hemolytic a... more vogeli, is a health problem in companion animals. The disease causes febrile illness, hemolytic anemia, pre-hepatic jaundice, and thrombocytopenia. Moreover, renal dysfunction from babesiosis has been reported. The purpose of this study was to investigate urinary markers that might be sensitive and specific for the early detection of renal dysfunction in B. c. vogeli-infected dogs. Blood and urine samples were collected from 11 dogs. The blood and urine samples were divided into two groups. B. c. vogeli-infected dogs group including six infected dogs as confirmed by microscopic examination and multiplex polymerase chain reaction. Non-infected dogs group included five healthy dogs. Blood samples were subjected to hematology and biochemistry analysis while urine samples were stored at -80 o C until analyzed. Three candidate urinary markers (urinary immunoglobulin G, uIgG; urinary C-reactive protein, uCRP; and urinary retinol-binding protein, uRBP) were examined using commercial enzyme-linked immunosorbent assays (ELISA); two additional candidate markers, aspartate aminotransferase to alanine aminotransferase ratio (AST/ALT) and urinary creatinine to serum creatinine ratio (UCr/SCr) were also studied. The results demonstrated that hemoglobin, red blood cell count, and hematocrit were significantly different between B. c. vogeli-infected dogs and non-infected dogs while the candidate markers were not. In conclusion, the selected candidate markers could not be used as urinary markers for renal dysfunction in B. c. vogeli-infected dogs. However, further study should investigate other urinary markers such as albumin, tubular enzymes, and tubular proteins as well as high-throughput technologies such as the proteomic approach.
The Southeast Asian Journal of Tropical Medicine and Public Health, 2014
Canine monocytic ehrlichiosis is a tick borne disease caused by Ehrlichia canis, an obligate intr... more Canine monocytic ehrlichiosis is a tick borne disease caused by Ehrlichia canis, an obligate intracellular rickettsial organism belonging to the family Anaplasmataceae. Canine ehrlichiosis causes hemaotological changes among infected animals which could be used as a potential predictor for diagnosing canine monocytic ehrlichiosis (CME). Ninety-four blood samples were obtained from canines that either presented for a routine health check-up or for clinical illness. A history, physical and laboratory test were conducted on each animal. All samples were examined for E. canis using a 16S rDNA polymerase chain reaction (PCR) amplification to confirm CME infection. Thirty-six of the samples were positive for E. canis using PCR and the rest were negative. The Mann-Whitney and chisquare test were used to compare the differences between the PCR-positive and negative animals. PCR-positive animals had a higher mean body temperature than PCR-negative animals. The following were significantly lower in PCRpositive animals: white blood cell count, eosinophil count, red blood cell count, hemoglobin, hematocrit, platelet count, and the random distribution of width (RDW) of the red blood cells. We evaluated complete blood cell count findings to determine factors associated with CME using multivariable logistic regression analysis and found thrombocytopenia was significantly associated with CME (OR=0.085; 95%CI: 0.78-0.92, p<0.001). For every decrease in the platelet count of 10,000 there was a 15% increase in the likelihood of having CME.
The Southeast Asian Journal of Tropical Medicine and Public Health, Jul 1, 2012
Canine ehrlichiosis is an endemic parasitic disease widely found in Thailand. The causative micro... more Canine ehrlichiosis is an endemic parasitic disease widely found in Thailand. The causative microorganism is tick-borne Ehrlichia spp, an obligate intracellular rickettsia residing in leukocytes. Ehrlichia spp in morulae-positive canine blood samples were identified using polymerase chain reaction amplification and direct sequencing of Ehrlichia spp. 16S rDNA 396 bp fragment and 36 of 59 were positive for E. canis. E. chaffeensis and E. ewingii were not detected. Sequencing alignment and phylogenetic analysis showed that 16S rDNA sequences of E. canis strains are 99.1-100% identical among E. canis strains from different countries worldwide. Further studies are required in order to determine new target sequence for genotyping of E. canis strains in the dog population in Thailand.
Sanprasert V, Sungpradit S, Nuchprayoon S. Wolbachia isolation by fractioned centrifugation from ... more Sanprasert V, Sungpradit S, Nuchprayoon S. Wolbachia isolation by fractioned centrifugation from Dirofilaria immitis for protein analysis. Chula Med J 2010 Objective The biological roles of Wolbachia, a filarial intracellular bacteria, have promoted applied researches in lymphatic filariasis. Analysis of Wolbachia proteins will provide complementary information on the biology of Wolbachia, as well as the immune mechanisms against filarial nematodes. We therefore developed a Wolbachia isolation method from Dirofilaria immitis (dog heartworm) for protein analysis. Methods Wolbachia bacteria were homogenized and isolated by fractioned centrifugation by using 0.85% NaCl supplemented with varying concentration of Nonidet P-40 (NP-40) as the homogenization buffer. The purity of Wolbachia extracts were analyzed by sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE), and by quantitative polymerase chain reaction (qPCR) targeting the single copy of the Wolbachia surface prot...
The Southeast Asian journal of tropical medicine and public health, 2014
Canine monocytic ehrlichiosis is a tick borne disease caused by Ehrlichia canis, an obligate intr... more Canine monocytic ehrlichiosis is a tick borne disease caused by Ehrlichia canis, an obligate intracellular rickettsial organism belonging to the family Anaplasmataceae. Canine ehrlichiosis causes hemaotological changes among infected animals which could be used as a potential predictor for diagnosing canine monocytic ehrlichiosis (CME). Ninety-four blood samples were obtained from canines that either presented for a routine health check-up or for clinical illness. A history, physical and laboratory test were conducted on each animal. All samples were examined for E. canis using a 16S rDNA polymerase chain reaction (PCR) amplification to confirm CME infection. Thirty-six of the samples were positive for E. canis using PCR and the rest were negative. The Mann-Whitney and chi-square test were used to compare the differences between the PCR-positive and negative animals. PCR-positive animals had a higher mean body temperature than PCR-negative animals. The following were significantly l...
The Southeast Asian journal of tropical medicine and public health, 2012
Canine ehrlichiosis is an endemic parasitic disease widely found in Thailand. The causative micro... more Canine ehrlichiosis is an endemic parasitic disease widely found in Thailand. The causative microorganism is tick-borne Ehrlichia spp, an obligate intracellular rickettsia residing in leukocytes. Ehrlichia spp in morulae-positive canine blood samples were identified using polymerase chain reaction amplification and direct sequencing of Ehrlichia spp. 16S rDNA 396 bp fragment and 36 of 59 were positive for E. canis. E. chaffeensis and E. ewingii were not detected. Sequencing alignment and phylogenetic analysis showed that 16S rDNA sequences of E. canis strains are 99.1-100% identical among E. canis strains from different countries worldwide. Further studies are required in order to determine new target sequence for genotyping of E. canis strains in the dog population in Thailand.
The Southeast Asian journal of tropical medicine and public health, 2012
Lymphatic filariasis, caused by filarial nematodes, is a mosquito-borne disease that affects over... more Lymphatic filariasis, caused by filarial nematodes, is a mosquito-borne disease that affects over 120 million people in the tropics and subtropics. The disease is caused mainly by Wuchereria bancrofti and Brugia malayi. Fertile adult female worms release offsprings (microfilariae) into the host blood circulatory system. Transmission-blocking agents as well as antimicrobial agents have been used to reduce microfilarial density in human and animal reservoir hosts. Doxycycline and rifampicin have an effect on the obligate intracellular gram-negative bacteria, Wolbachia, which appears to exert an influence on filarial nematode embryonic and larval development, adult female fertility, and filarial survival. We investigated the effects of doxycycline, rifampicin and ciprofloxacin on B. malayi microfilarial motility, expressed as minimum effective concentration (MEC), and on Wolbachia proliferation using quantitative PCR, expressed as the concentration of the drug to inhibit bacteria growt...
Asian Pacific Journal of Tropical Disease, 2012
International Journal of Infectious Diseases, 2010
Background: Prevalence of chloroquine resistant malaria is on a rise and our area is one of the d... more Background: Prevalence of chloroquine resistant malaria is on a rise and our area is one of the declared endemic zones for malaria. Recent mortality trends of the disease have increased considerably seeking immediate modification in the treatment guidelines to decrease the complications and thus the mortality of the disease. We have attributed the present condition to the chloroquine resistance, the drug which is used to treat the disease in this area for so long. Even the effective surveillance system fails in decreasing the mortality figures by following the prescribed treatment guidelines. Hence, we have undertaken this project to assess the drug resistance and to state new treatment guidelines in the areas where chloroquine resistant malaria is rampant.
Forensic Science International, 2010
Parasitology international, Jan 21, 2015
Beef cattle production represents the largest cattle population in Thailand. Their productivity i... more Beef cattle production represents the largest cattle population in Thailand. Their productivity is constrained by tick-borne diseases such as babesiosis and theileriosis. In this study, we determined the prevalence of Babesia bigemina, Babesia bovis and Theileria orientalis using polymerase chain reaction (PCR). The genetic markers that were used for detection of the above parasites were sequenced to determine identities and similarity for Babesia spp. and genetic diversity of T. orientalis. Furthermore the risk factors for the occurrence of the above protozoan parasites in beef cattle from northern and northeastern parts of Thailand were assessed. A total of 329 blood samples were collected from beef cattle in 6 provinces. The study revealed that T. orientalis was the most prevalent (30.1%) parasite in beef cattle followed by B. bigemina (13.1%) and B. bovis (5.5%). Overall, 78.7% of the cattle screened were infected with at least one of the above parasites. Co-infection with Babes...
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Papers by Sivapong Sungpradit
Materials and Methods: The study included 33 B. vogeli–positive blood samples and 11 E. canis–coinfected blood samples. To exclude coinfection with Hepatozoon canis and Anaplasma platys, only dogs with B. vogeli infection and B. vogeli–E. canis coinfection were included in the study. A multiplex polymerase chain reaction (PCR) assay was conducted to detect B. vogeli, E. canis, and H. canis, and a conventional PCR assay was conducted for the detection of A. platys. Besides, the PCR assay and sequencing, comprehensive data analysis was conducted, including a microscopic blood parasite examination and hematological and biochemical data analysis.
Results: The comparison of the hematobiochemical data between the B. vogeli–positive and E. canis coinfection groups identified that there were statistically significant differences in the RBC parameters, including RBC count, hemoglobin concentration, hematocrit, and RBC distribution width (p=0.001). Neither B. vogeli infection nor coinfection with E. canis was associated with the sex, breed, recorded clinical signs, geographic origin of the dog and also B. vogeli 18S rRNA gene sequencing results.
Conclusion: Coinfection with E. canis increased the severity of babesiosis. The pathogenic mechanisms underlying this infection, such as destruction of RBCs, require further investigation. This study may enhance diagnosis, treatment, and prevention of canine babesiosis.
Materials and Methods: The study included 33 B. vogeli–positive blood samples and 11 E. canis–coinfected blood samples. To exclude coinfection with Hepatozoon canis and Anaplasma platys, only dogs with B. vogeli infection and B. vogeli–E. canis coinfection were included in the study. A multiplex polymerase chain reaction (PCR) assay was conducted to detect B. vogeli, E. canis, and H. canis, and a conventional PCR assay was conducted for the detection of A. platys. Besides, the PCR assay and sequencing, comprehensive data analysis was conducted, including a microscopic blood parasite examination and hematological and biochemical data analysis.
Results: The comparison of the hematobiochemical data between the B. vogeli–positive and E. canis coinfection groups identified that there were statistically significant differences in the RBC parameters, including RBC count, hemoglobin concentration, hematocrit, and RBC distribution width (p=0.001). Neither B. vogeli infection nor coinfection with E. canis was associated with the sex, breed, recorded clinical signs, geographic origin of the dog and also B. vogeli 18S rRNA gene sequencing results.
Conclusion: Coinfection with E. canis increased the severity of babesiosis. The pathogenic mechanisms underlying this infection, such as destruction of RBCs, require further investigation. This study may enhance diagnosis, treatment, and prevention of canine babesiosis.