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Studi virulensi trypanosoma evansi isolat indonesia dengan penentuan marka molekular DNA mikrosatelit dan analisis profil sitokin pada mencit Mus musculus = Virulence study of trypanosoma evansi isolates from Indonesia and identification of molecular marker based on microsatellite DNA and cytokine profile analyses in mice Mus musculus

Dyah Haryuningtyas Sawitri; Mohamad Sadikin, supervisor; Heri Wibowo, co-promotor; April Hari Wardhana, co-promotor; Indra Gusti Mansur, examiner; Taniawati Supali, examiner; Dwi Ari Pujianto, examiner; Dondin Sajuthi, examiner ([Publisher not identified] , 2016)

 Abstrak

ABSTRAK
Pendahuluan :Trypanosoma evansi adalah protozoa berflagella yang bersirkulasi
di dalam darah secara ekstraseluler sebagai agen penyakit Surra serta menyerang
seluruh hewan vertebrata, serta berpotensi sebagai zoonosis. Informasi virulensi
isolat T. evansi sangat dibutuhkan untuk penentuan strategi pengobatan Surra di
daerah wabah dan endemis. Penelitian ini bertujuan untuk mengetahui variasi
virulensi isolat T. evansi yang dikoleksi dari berbagai wilayah di Indonesia
termasuk memperoleh marka genetik serta mengetahui profil sitokin pada mencit.
Disamping itu, dilakukan juga uji serologis pada peternak di daerah wabah Surra
Metode : Sebanyak 32 isolat lokal T. evansi dikonfimasi dengan PCR multiplex
(ITS-1; Te Ro Tat 1,2 VSG dan ESAG6/7), selanjutnya diuji virulensinya dengan
menginfeksikan104 parasit pada mencit galur DDY. Studi genotyping populasi
T. evansi dievaluasi dengan 8 marka mikrosatelit Tbb-1, Tbb-5, Tbb-9, Tbb-10,
MORF2-CA, M6C8-CA, MEST-19AT, MT3033-AT. Dua isolat yang berbeda
virulensi (tinggi-Bang87 dan rendah-Pml 287) dipilih untuk uji imunopatogenitas
sedangkan serum peternak diuji dengan metode FELISA dan CATT T. evansi.
Hasil : Dari 32 isolat tersebut terbagi menjadi 17 isolat bervirulensi tinggi, 11
isolat bervirulensi moderat dan 4 isolat bervirulensi rendah dengan 8 pola tingkat
parasitemia. Analisis Neigbour Joining (NJ) terhadap 8 lokus berdasarkan Multi
Lokus Genotipe (MLG) mikrosatelit terbagi menjadi 4 populasi, yaitu MLG A,
MLG B, MLG C dan MLG D. Analisis terhadap struktur populasi juga
memberikan hasil yang sama dengan terbentuknya 4 klaster. Hasil ini juga
membuktikan bahwa marka yang digunakan bersifat spesifik lokasi. Sebanyak
tiga marka mengindikasikan adanya asosiasi antara virulensi dan MLG (Tbb-1,
M6C8-CA dan MEST-19). Kadar IFN-γ meningkat secara tajam pada mencit
yang diinfeksi isolat Bang 87 pada 4hpi berkorelasi negatif yang signifikan
(p<0,05) dengan kadar IL-10, sedangkan pada mencit yang diinfeksi isolat Pml
287, peningkatan kadar IFNγ berkorelasi positif dengan kadar IL-10. Kematian
dini pada mencit yang diinfeksi isolat Bang 87 disebabkan oleh sindrom respon
inflamasi sitemik. Hasil uji serologis menunjukkan bahwa 4 dari 24 serum
peternak (16,67%) didaerah wabah positif dan seluruh serum negatif untuk
daerah non wabah.
Kesimpulan : Variasi virulensi T. evansi isolat Indonesia memiliki karakter
molekular yang berbeda serta menginduksi pola mediator sitokin pro dan
antiinflamasi yang berhubungan dengan pola manifestasi patologi yang berbeda.
Marka mikrosatelit pada studi ini mampu mengidentifikasi asal usul sumber
infeksi, dan tingkat virulensi isolat yang sedang bersirkulasi. Surra berpotensi
sebagai emerging zoonosis, terutama bagi peternak didaerah wabah dan endemis.
ABSTRACT
Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas;Introduction: Trypanosoma evansi is an extracellular homoflagellate of
protozoan blood causing Surra. The disease attacks all vertebrates and potentially
as zoonosis. Virulence analysis of T. evansi is a fundamental knowledge to
determine treatment strategies of Surra in both outbreak and endemic areas. The
aims of this study was to determine virulence variation of T. evansi isolates
collected from various regions in Indonesia and to obtained genetic markers as
well as cytokine profile in mice. In addition, serological test was also carried out
to farmers living in a Surra outbreak area
Methods: Total of 32 isolates of T. evansi corfirmed with multiplex PCR (ITS-1;
Te Ro Tat 1.2 VSG and ESAG6 / 7), were further tested with inoculation 104
parasite in DDY mice strain. The population genotype study of T. evansi was
evaluated with 8 microsatellite markers (Tbb-1, Tbb-5, Tbb-9, Tbb-10-CA
MORF2, M6C8-CA, MEST-19AT, MT3033-AT). Two different virulence
isolates, high-Bang87 and low-PML287 was selected to cytokine profile analysis
using ELISA, while farmers sera were tested using CATT and FELISA kits
Results: A total of 32 local isolates of T. evansi tested were divided into three
different virulences, i.e. 17 high virulence isolates, 11 moderate virulence and 4
low virulence isolates forming 8 pattern parasitemia levels. Based on Neigbour
Joining (NJ) on 8 Microsatellite Multilocus Genotype (MLGs) was grouped into
4 populations (MLG A, MLG B, MLG C and MLG D). Stucture population
analysis also provided the similar result generating 4 clusters. These results
indicated that the markers used in this study had a specific location property.
Three markers (TBB-1, and MEST M6C8-CA-19) showed an association
between virulence and MLG. IFN-γ levels increased significantly in mice
infected with Bang 87 isolate on 4th day post infection (dpi) having a significant
negative correlation (p <0.05) with increased IL-10 levels, whereas in mice
infected by PML 287 isolate, IFN-γ levels were positively correlated with IL- 10
levels. Early death in mice infected with Bang87 isolates was caused by systemic
inflammatory response syndrome (SIRS). Result of serological test showed that 4 out
of 24 farmers sera (16.67%) from outbreak areas are positive and all sample from
free area are negative.
Conclusion: Virulence variation of T. evansi isolates from Indonesia has
different molecular character and induces cytokine pattern of pro and antiinflammatory
mediators associated with distinct patterns of pathological
manifestations. The microsatellite markers found in this study are able to identify
origin of infection sources dan determine virulence of isolates that circulate on the
outbreak area. Surra is potential new emerging disease, particularly for farmers or
immunosurpressed individuals who living in both endemic and outbreak areas

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Penerbitan : [Place of publication not identified]: [Publisher not identified], 2016
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Deskripsi Fisik : xxii, 136 pages : illustration ; 28 cm + appendix
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