アトピー性皮膚炎から分離された黄色ブドウ球菌が産生するエンテロトキシンSEYは特異的なTCRVαレパートリーのT細胞を活性化させる

内容の要約広島大学(Hiroshima University)博士(学術)Doctor of Philosophydoctora

Genetic Determination and Clonal Relationships of Staphylococcus aureus Isolated from Dairy Cows in Baturraden, Central Java, Indonesia

Cases of mastitis in cows at Baturraden are leading to signifi cant and ongoing problems due to reducedproduction and lower milk quality. This study was designed to identify which of selected virulence determinantgenes of S. aureus are involved in the Baturraden infection, and to determine the clonal relationship amongthese isolates. Seventeen isolates were identifi ed as S. aureus based on their biochemical properties and speciesspecifi city for 23S rRNA and nuc genes. S. aureus isolates were genotypically characterized for the selectedvirulence determinants: coa, clfA, fnbA, fnbB, cap5, spa IgG and spa X- region genes. Clonal relationship analysisamong isolates was carried out using AFLP and results compared with previously confi rmed relationshipsbetween selected S. aureus isolated from other regions. The results show that eight isolates contain all thegenes, but six isolates lack fnbB and two isolates lack cap5 genes. AFLP analysis showed that all isolates of S.aureus originating from cows in Baturraden belong to one cluster. This study provides additional knowledgeabout S. aureus infection in Baturraden cows, including the number of virulence determinant genes that mayplay a role in pathogenicity

Genetic Determination and Clonal Relationships of Staphylococcus aureus Isolated from Dairy Cows in Baturraden, Central Java, Indonesia

Cases of mastitis in cows at Baturraden are leading to signifi cant and ongoing problems due to reducedproduction and lower milk quality. This study was designed to identify which of selected virulence determinantgenes of S. aureus are involved in the Baturraden infection, and to determine the clonal relationship amongthese isolates. Seventeen isolates were identifi ed as S. aureus based on their biochemical properties and speciesspecifi city for 23S rRNA and nuc genes. S. aureus isolates were genotypically characterized for the selectedvirulence determinants: coa, clfA, fnbA, fnbB, cap5, spa IgG and spa X- region genes. Clonal relationship analysisamong isolates was carried out using AFLP and results compared with previously confi rmed relationshipsbetween selected S. aureus isolated from other regions. The results show that eight isolates contain all thegenes, but six isolates lack fnbB and two isolates lack cap5 genes. AFLP analysis showed that all isolates of S.aureus originating from cows in Baturraden belong to one cluster. This study provides additional knowledgeabout S. aureus infection in Baturraden cows, including the number of virulence determinant genes that mayplay a role in pathogenicity

Deteksi Staphylococcus aureus dan Staphylococcus sp. Secara Langsung Dari Susu Segar Kambing Peranakan Etawa dengan Teknik PCR

Genus Staphylococcus merupakan salah satu patogen bakteri penyebab mastitis yang menyebabkan kerugian ekonomi pada kambing Peranakan Etawa. Diantara Staphylococcus sp. yang dapat tumbuh dengan baik dalam susu segar, diketahui Staphylococcus aureus dapat membahayakan kesehatan manusia yang mengkonsumsi (food borne disease) karena kemampuannya dalam memproduksi enterotoksin yang tahan terhadap enzim pencernaan maupun pemanasan. Tujuan penelitian ini adalah mendeteksi Staphylococcus sp. dan S. aureus secara langsung dari susu kambing peranakan etawa dengan teknik PCR.Metode yang dilakukan adalah mengekstraksi DNA dari 60 sample susu segar dengan prinsip spin column-based nucleic acid purification dan kemudian dilakukan amplifikasi gen spesifik 23S rRNA Staphylococcus sp. dan S. aureus. Hasil PCR diketahui 37 (61%) sampel susu positif mengandung Staphylococcus sp. dan hanya 1 (1,6%) sampel mengandung S. aureus. Metode deteksi dengan PCR dapat digunakan untuk mendeteksi kontaminan Staphylococcus sp. dan S. aureus dengan waktu yang singka

Identifikasi Molekuler Bakteri Staphylococcus sp. dan Staphylococcus aureus Penyebab Mastitis Subklinis pada Ternak Kambing Perah

Mastitis merupakan radang pada glandula mammae (ambing) ternak perah. Mastitis tipe subklinis sering dikaitkan pada kejadian mastitis di peternakan ruminansia kecil seperti kambing perah (kambing Peranakan Etawah, Saanen, dan Sapera). Patogen utama yang berperan dalam kejadian mastitis ini adalah genus Staphylococcus. Tujuan penelitian adalah untuk melakukan identifikasi bakteri Staphylococcus sp. dan Staphylococcus aureus sebagai penyebab mastitis subklinis pada kambing perah dengan menggunakan metode polymerase  chain reaction (PCR). Tahapan metode yang dilakukan adalah ekstraksi DNA dengan teknik spin-collumn system terhadap 26 isolat bakteri yang telah dilakukan uji biokimia sebelumnya dan amplifikasi gen spesifik 23s rRNA Staphylococcus sp. dan Staphylococcus aureus, serta methicillin resistant Staphylococcus aureus (MRSA), dilanjutkan dengan visualisasi menggunakan UV-transluminator. Hasil menunjukkan bahwa sebanyak 12 isolat sampel teridentifikasi Staphylococcus sp. dan 1 diantaranya teridentifikasi Staphylococcus aureus. Isolat yang teridentifikasi Staphylococcus aureus bukan termasuk MRSA.

Deteksi Staphylococcus aureus dan Staphylococcus sp. secara Langsung dari Susu Segar Kambing Peranakan Etawa dengan Polymerase Chain Reaction (PCR)

The genus of Staphylococcus is one of the bacterial pathogens that cause mastitis causing economic losses in Etawa crossbreed goat. Among Staphylococcus sp. which can grow well in raw milk, Staphylococcus aureus is known causing foodborn disease in human because of its ability to produce enterotoxins that are resistance to digestive enzymes or heating treatment. The purpose of this study was to detect Staphylococcus sp. and S. aureus directly from raw milk of Etawa crossbreed goat using PCR. The study was carried out by extracted DNA from fresh milk using spin column method and then amplified specific 23S rRNA for Staphylococcus sp. and S. aureus. PCR examination revealed that 37 (61%) and 1 (1,6%) raw milk samples were positive for Staphylococcus sp. and S. aureus respectively. The PCR Method is usefull to reduce defection time of Staphylococcus sp. contaminants

Komparasi Lima Jenis Primer Polymerase Chain Reaction Untuk Mengidentifikasi Kelamin Burung Famili Columbidae Yang Akurat

Penentuan jenis kelamin pada beberapa spesies burung cukup sulit dilakukan dikarenakan jantan dan betina memiliki ciri morfologi yang sama (monomorfik), salah satunya famili Columbidae. Teknik penentuan jenis kelamin burung secara molekuler yang populer adalah metode Polymerase Chain Reaction (PCR) dengan gen target Chromo Helicase DNA-binding (CHD), namun keberhasilan amplifikasi gen target pada PCR tersebut dipengaruhi salah satunya kesesuaian DNA template dengan primer yang digunakan. Penelitian ini bertujuan untuk mengevaluasi 5 jenis primer PCR P2/P8, 2550F/2718R, CHD1F/CHD1R, 1237L/1272H dan CHD1LF/CHD1LR untuk menentukan jenis kelamin famili Columbidae. Penelitian ini dilakukan dengan menguji 5 jenis primer tersebut di atas pada sampel DNA tiap pasang jantan dan betina dari burung merpati, balam jambi, punai, derkuku, dan perkutut. Hasil penelitian diketahui primer CHD1LF/CHD1LR menunjukkan hasil terbaik dan direkomendasikan untuk menentukan jenis kelamin famili Columbidae

Deteksi Gen Penyandi Sifat Resistensi Metisilin, Penisilin Dan Tetrasiklin Pada Isolat Staphylococcus Aureus Asal Susu Mastitis Subklinis Sapi Perah

Detection of gene encoding resistance of bacteria could be used as an accurate method to determine resistance of Staphylococcus aureus which is causing mastitis in dairy cows to the several antibiotics. This research aimed to detect the gene encoding resistance of methicillin, penicillin and tetracycline from identified S. aureus. Sixty milk samples were collected from subclinical mastitis of cows from various dairy farming in Yogyakarta. Isolation and identification of S. aureus based on the culture, Gram staining and biochemical test. Phenotypes of S. aureus resistances against antibiotics were carried out by disc diffusion method, meanwhilespecies specific gene of S. aureus and the gene encoding methicillin, penicillin and tetracycline were confirmed by PCR method. The results showed 11 isolates representing of Methicillin Susceptible Staphylococcus aureus (MSSA) could be identified, wherein 5 isolates were harboring both of penicillin and tetracycline resistant genes respectively

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Birds sexing utilize the Polymerase Chain Reaction (PCR) technique is increasingly being used by researchers and breeders. The PCR technique has high sensitivity, but its success is influenced by the specificity of the DNA template with the oligo primer used. This study aimed to evaluate 5 types of PCR primers P2/P8, 2550F/2718R, CHD1F/CHD1R, 1237L/1272H, and CHD1LF/CHD1LR to determine the sex of Phasianidae, Anatidae, Muscicapidae, and Psittacidae families. This research was conducted by tested primers mentioned above to amplify the target gene chromodomain helicase DNA binding 1 (CHD1) on DNA samples of each pair of males and females from four bird families, respectively. The results indicated that CHD1LF/CHD1LR PCR primer gave the best results and was recommended to determine the sex of four families tested. Some of other primers tested in this study failed to amplify targeted gene correctly, it is important to use appropriate primer to increase bird sexing accuracy

Deteksi Gen Penyandi Sifat Resistensi Metisilin, Penisilin dan Tetrasiklin pada Isolat Staphylococcus aureus Asal Susu Mastitis Subklinis Sapi Perah

Detection of gene encoding resistance of bacteria could be used as an accurate method to determine resistance of Staphylococcus aureus which is causing mastitis in dairy cows to the several antibiotics. This research aimed to detect the gene encoding resistance of methicillin, penicillin and tetracycline from identified S. aureus. Sixty milk samples were collected from subclinical mastitis of cows from various dairy farming in Yogyakarta. Isolation and identification of S. aureus based on the culture, Gram staining and biochemical test. Phenotypes of S. aureus resistances against antibiotics were carried out by disc diffusion method, meanwhilespecies specific gene of S. aureus and the gene encoding methicillin, penicillin and tetracycline were confirmed by PCR method. The results showed 11 isolates representing of Methicillin Susceptible Staphylococcus aureus (MSSA) could be identified, wherein 5 isolates were harboring both of penicillin and tetracycline resistant genes respectively