PERBANDINGAN ANTARA ALANTOIN (5 UREIDOHYDANTOIN) DENGAN BETADINE® (POVIDONE IODINE) UNTUK PENGOBATAN LUKA BVSISI

Study on the comparison between allantoin (5 ¢ ureidohydantoin) and Betadine ® (povidone iodine) was conducted to compare and evaluate their efficacy, especially to accelerate wound (incision) healing. Treatment divided into three groups, first group is Control (without therapy), second group is allantoin treatment and the last one is Betadine ® treatment. Allantoin obtained from cattle's urine by Meissner method. The solution made of 2,4 grams of allantoin in 600 milliliters aqueous solution. Treatments (therapies) were given three times a day topically. Results showed no significant difference between allantoin and Betadine ® treatments (p > 0,05), control and the other treatments i.e allantoin and Betadine ® therapies have significantly difference (p < 0,01)

HUBUNGAN TINGKAT KELAHIRAN DAN KEMATIAN DENGAN TINGKAT KEPADATAN POPULASI RUSA BAWEAN DI KEBUN BINATANG SURABAYA

Penelitian ini bertujuan untuk mengetahui bentuk dan derajat hubungan yang terjadi antara tingkat kelahiran dan kematian populasi Rusa Bawean dengan tingkat kepadatannya

POLYCLONAL ANTIBODY UTILIZATION FOR DETECTION OF JEMBRANA ANTIGEN AT BALI CATTLE IN WEST SUMATRA PROVINCE

The aims of this study were to detect Jembrana antigen with polyclonal antibody and to describe antigen distribution in the Bali cattle organ that positively infected with Jembrana disease. Spleens, lungs, and livers were harvested from 10 naturally infected Bali cattle whose infection was confirmed through positive Polymerase Chain Reaction (PCR) result for Jembrana virus. Immunohistochemistry test was performed using polyclonal antibodies produced in rabbits. The results showed that cells infected by Jembrana viruses displayed positive reaction with a reddish brown color. Immunohistochemistry methods using polyclonal antibodies can detect Jembrana antigens in the spleen, liver, and lung with the highest average detection score (P0.05) was found in spleen, followed by liver and lungs. There was significant difference in the distribution of Jembrana antigens between the spleen, liver, and lungs with spleen having the highest antigen density

Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate

Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important roleduring cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccinecandidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encodingMIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction withspecific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue byheat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestionusing restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequencedto find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence thenwere analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue bypGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5&rsquo; and 447 from 3&rsquo; of genesequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate.Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DN

Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate

Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important role during cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccine candidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encoding MIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction with specific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue by heat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestion using restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequenced to find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence then were analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue by pGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5’ and 447 from 3’ of gene sequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate. Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DN

Prevalence of Trypanosomiasis of Wild Rats (Rattus sp.) in Banjarnegara District and Potential Impact for Public Health

Trypanosomiasis is a zoonotic disease caused by Trypanosoma sp., a protozoan parasite that has a flagellum. It has the potential to cause emerging diseases. Generally, Trypanosoma infection is caused by T. evansi which causes Surra disease, and T. cruzi which causes Chagas disease. Trypanosoma lewisi has been considered a natural protozoan in mice, not pathogenic to humans but in recent years it has been reported in humans. This study aims to detect Trypanosoma in rats in Banjarnegara District and analyze the potential impact on public health. The research was observational with a descriptive approach, conducted in Banjarnegara from July-December 2020. Samples were taken by purposive sampling. Samples are rat’s blood that caught on wild rats survey in the main market of Banjarnegara District. Blood samples were made with a thin smear then they were stained with Giemsa and examined by microscope. There were 157 rats caught, consisting of 131 Rattus norvegicus and 26 R. tanezumi. Totally, 28 rats were positive Trypanosoma lewisi, so Trypanosoma infection in rats in Banjarnegara District is 16,57%. Trypanosomiasis in R. norvegicus was 18.3% and R. tanezumi 15.38%. Therefore, there is a need to increase the awareness of these diseases’s transmission to humans

Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate

Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during activepenetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targetedcell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasitesuccessfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorusvacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently,this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone andsequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique.Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA wasused as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor fromRiboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinantplasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agarcontaining X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in theLB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order toidentify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolatedusing alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid wascut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward andM13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretoryand secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the clonedgene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.Keywords: Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP

Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate

Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during active penetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targeted cell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasite successfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorus vacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently, this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone and sequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique. Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/ c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA was used as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor from Riboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinant plasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agar containing X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in the LB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order to identify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolated using alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid was cut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward and M13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretory and secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the cloned gene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate. Keywords: Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP

EVALUATION OF B1 GENE TO DETECT Toxoplasma gondii: COMPARISON OF THREE SETS NESTED PCR PRIMER

This study aimed to evaluate three sets of B1 gene DNA primer for the diagnosis of Toxoplasma gondii. The DNA of Toxoplasma gondii that  stored on liquid nitrogen was isolated using DNAzol™ reagent. The first step of Polymerase Chain Reaction (PCRs) was performed using external and internal primer sets, respectively, and then nPCR. PCR products sequencing was performed by Apical Science. All sequences were analysed using CLC Sequence Viewer Version 8.0 software and compared to sequence database that deposited in ToxoDB (Toxoplasma gondii genome database) using BLAST (https://toxodb.org/toxo/app). Each B1 gene primer was evaluated by performing single PCR (forward and reverse) and nested PCR reactions. Three sets of B1 gene primer have different amplification precision. According to the results of amplicon sequencing, the primer set #2 has the best amplification precision of B1 gene