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PROTOTIPE KTT DIAGNOSTIK UNTUK DETEKSI STREPTOKOKOSIS PADA PRIMATA DENGAN ELIS A-ANTIBODI MONOKLONAL PENANGKAP ANTIGEN Siti Isrina Oktavia Salasia; Aris Purwantoro; . Khusnan
Jurnal Sain Veteriner Vol 25, No 1 (2007): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Penelitian ini bertujuan mengembangkan sarana diagnostik untuk kontrol streptococcosis pada primata dengan ELISA-monoclonal antibody penangkap antigen M-like protein (MLP) Streptococcus equi subsp. zooepidemiats grup C (SGC). M-like protein SGC diekstraksi dengan menggunakan lisozim dan Nasetilmuramidase. Protein dengan besar sekitar 58 kDa digunakan sebagai antigen untuk menimbulkan antibodi pada mencit Balb/c. Mencit yang mengandung antibodi dengan absorban tertinggi (2,868) diambil limpanya untuk memperolehl imfosit imUn( limfoblast). Hasil fusi sel mieloma dan limfoblast diperoleh4 klon hibridoma yang positif mengandung antibodi terhadap MLP, dengan nilai absorban pada ELISA masing-masing 1,900, 1,963, 1,895 dan 2,050. Hasil propagasi cairan asites mencit Balb/c diperoleh monoklonal antibodi terhadapMLP S. equi subsp. zooepidemicus dengan nilai absorban dan konsentrasi sebagai berikut: asitesI = 1,597(5,50mg), asites 2 : 1,940 (5,75 mg), dan asites 3 : 3,012 (5,80 mg). Antibodi monoklonal memperlihatkanspesifitas yang cukup tinggi karena hanya mengenal I epitop spesifik yang diperlihatkan pada uji Western blot dengan menampakkan pita tunggal pada sekitar 58 kDa dan menunjukkan reaksi positif pada uji dot-blot. Antibodi monoklonal memperlihatkan sensitifitas yang cukup tinggi setelah diuji dengan serum hewan percobaan tikus yang diinfeksi buatan dengan S. equi subsp. zooepidemicus dengan hasil absorban pada uji ELISA lebih dari 1,00 dan menunjukkan reaksi positif pada uji dot-blot. Hasil uji terhadap sampel serum Macacafascicularis menunjukkan bahwa 97 ,56% positif.Kata kunci: S. equi subsp. zooepidemicus, antibodi monoklonal, M-like protein, primata

Respon Neutrofil, Adesi Pada Sel Epitel, Aglutinasi Eritrosit Terhadap Staphylococcus aureus : Kajian Hidrofobisitas In Vitro = Response of neutrophils, epithelial cells adhesion, erythrocytes agglutination of Staphyloco Khusnan .; Siti Isrina Oktavia Salasia
Jurnal Sain Veteriner Vol 24, No 1 (2006): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Staphylococcus aureus merupakan salah satu bakteri potensial sebagai penyebab utama mastitis pada sapi perah. Mastitis dapat menyebabkan kerugian ekonomi peternak akibat turunnya produksi susu. Infeksi bakteri dapat terjadi melalui kemampuan bakteri memasuki hospes, berkembang biak, merusak jaringan inang dan mampu bertahan dalam tubuh hospes. Penelitian ini bertujuan untuk mengetahui hubungan sifat hidrofobisitas S. aureus dan kemampuannya terhadap aglutinasi eritrosit, pelekatan dengan sel epitel dan kemampuan bertahan terhadap fagositosis sel polimorfonuklear. Dari 10 isolat S. aureus yang digunakan dalam penelitian ini, terdapat 8 isolat bersifat hidrofob dan 2 isolat bersifat hidrofil. Diantara isolat yang bersifat hidrofob terdapat 2 isolat mempunyai kemampuan mengaglutinasi eritrosit sapi perah, kambing, domba. Staphylococcus aureus yang bersifat hidrofob dan hemaglutinasi positif, lebih banyak melekat pada sel-sel epitel bukalis dan lebih banyak difagosit oleh sel-sel PMN dibanding isolat yang bersifat hidrofob tetapi hemaglutinasi negatif maupun isolat yang bersifat hidrofil. Isolat yang bersifat hidrofil tidak mampu mengaglutinasi eritrosit dan lebih sedikit melekat pada sel-sel epitel dan lebih sedikit difagosit oleh sel-sel PMN.

KARAKTERISASI FENOTIPE Streptococcus suis = THE PHENOTYPE CHARACTERIZATION OF Streptococcus suis Khusnan .; Siti Isrina Oktavia S.
Jurnal Sain Veteriner Vol 19, No 2 (2001): DESEMBER
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.381

Streptococcus suis telah diketahui menyebabkan meningitis, artritis dan septikemia pada babi, manusia dan hewanhewan lain. Penelitian ini bertujuan untuk mengetahui sifat fenotipe Streptococcus sins rneliputi sifat pertumbuhannyi dalam media cair, media soft agar, morfologi koloni dan sifat pelekatannya dalam uji heksadekan. Dari 34 isolat Streptococcus suis yang digunakan dalam penelitian ini terdapat 5 isolat tumbuh jernih dalam media cair dengan sedimen tampalc pada dasar tabung, tumbuh kompak dalam media soft agar, dengan pewarnaan methylene blue koloni membentuk rantai panjang dan berdasar uji heksadekan bersifat hidrofob. Kultur Streptococcus suis yang lain (29 isolat) tumbuh keruh dalam media cair, tumbuh difus dalam media soft agar, membentuk koloni tunggal atau rantai pendek dan bersifat hidrofil.

Karakterisasi Faktor-faktor Virulensi Staphylococcus aureus Asal Susu Kambing Peranakan Ettawa secara Fenotip dan Genotip Khusnan Khusnan; Wahyu Prihtiyantoro; Hartatik Hartatik; Mitra Slipranata
Jurnal Sain Veteriner Vol 34, No 1 (2016): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Staphylococcus aureus is a major cause of mastitis in large or small ruminants, and often manifested by subclinical mastitis in Peranakan Ettawa (PE) goats. Staphylococcus aureus in human can cause food borne disease. The research aimed to characterize the virulence factors of Staphylococcus aureus isolated from milk PE goats, phenotypic- and genotypically. Phenotypically characterization were determined through the pigmen assay as well as hydrophobicity, haemolysin, and hemaglutinin reaction. Polymerase chain reaction (PCR) analysis was used to detect 4 virulen genes including coa, clf, fnbA, and fnbB genes. The results of research showed that Staphylococcus aureus abled to produce white pigmen (35,7%), yellow pigmen g (57,1%), andorange pigmen (7,2%). Staphylococcus aureus showed α-hemolysis zone (35,7%), β-hemolysis (35,7%), dan γ-hemolysis (28,9%). Hydrophobicytic test revealed 14,3% Staphylococcus aureus isolates were hydrophobe and85,7% hydrophil. Staphylococcus aureus (85,7%) isolates abled to aglutinated sheep blood cells. Based on genotypic analysis of Staphylococcus aureus could be detected coa gene (92,8%), clf gene (64,3%), fnbA gene (78,6%), and fnbB gene (64,3%). Based on the phenotypic and genotypic characters, it can be concluded that Staphylococcus aureus are virulent strains. This information can be used as the basis for control mastitis in PE goats

Deteksi Hemaglutinin, Hemolisin dan Koagulase Secara Fenotipik dan Genotipik pada Staphylococcus aureus Isolat Asal Broiler Khusnan Khusnan; Dwi Kusmanto
Jurnal Sain Veteriner Vol 36, No 1 (2018): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Staphylococcus aureus is a bacterium causing diseases in animals and human. Staphylococcus aureus in broilers cause septicemia, tendosinovitis, dermatitis, endocarditis, wound infections, arthritis and bumblefoot. In this research, 24 isolates of Staphylococcus aureus from broiler were characterized of its virulent factors including the presence of haemagglutinin, ability to agulate the plasma in tubular coagulase test as well presipitate formation in clumping factor test, and haemolysis types. By polymerase chain reaction (PCR) were genotypically detected genes coa, clf, hlaA, and hlaB. All of the isolates (100%) had haemagglutinin, capable to agglutinate and precipitae of rabbit plasma. All isolates could lyse sheep red blood cells with the type of α-hemolysis (45.8%), β-hemolysis (50.0%) and γ-haemolysis (4.2%). Genotypically, all isolates (100%) had coa and clf genes, hlaA gene 7(0.8%) and hlaB gene (29.2%).

Prevalensi Strain Avian Pathogenic Escherichia coli (APEC) Penyebab Kolibasilosis pada Burung Puyuh Wahyu Prihtiyantoro; Khusnan Khusnan; Mitra Slipranata; Imron Rosyidi
Jurnal Sain Veteriner Vol 37, No 1 (2019): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Avian Pathogenic Escherichia coli (APEC) is a pathogen that causes colibacillosis in poultry, including salpingitis, omphalitis, cellulitis, swollen head syndrome, coligranuloma yolk sac inflammation, and air sacs inflammation. APEC is a zoonotic strain which spread through raw meat and processed meat products of animals and birds. In this research, the isolation and identification of Escherichia coli were done by using selective media MacConkey, Kligger Iron Agar, and Gram staining. Polymerase chain reaction (PCR) was used to analyse genetopically to detect 16SrRNA genes, vt1 genes, and vt2 genes. Thirty one (55,36%) isolates of 56 specimens collected from quail were detected as Escherichia coli. The detection of APEC strains towards 31 Escherichia coli isolates were done by using polymerase chain reaction (PCR) with vt1 and vt2 specific primer. The results showed that 32,26% (10/31) was APEC strains and 67.74% was non-APEC strains. From 10 isolates, 90% had vt1 gene and 10% had vt2 gene. Escherichia coli isolates were found in eyes (32,26%), infraorbital sinus fluid (32,26%), nasal fluid (16,20%), also in lungs, air sacs, ascites, and heart for 3,2% each. The isolates could not be found in the specimens from the skull. As a zoonotic agent, the isolates have an impact on human health.

Peran Hemaglutinin dan Hemolisin pada Escherichia coli Sorbitol-negatif Isolat Burung Puyuh pada Proses Infeksi Secara in Vitro Khusnan Khusnan; Wahyu Prihtiyantoro; Dwi Kusmanto
Jurnal Sain Veteriner Vol 39, No 3 (2021): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.69896

Abstract Escherichia coli sorbitol-negative in quails cause economic loss due to the death, growth rate inhibition, decreased egg production, and increased medical treatment. Escherichia coli sorbitol-negative has many virulence factors, including hemagglutinin and hemolysin. The aim of this study is to determine the role of hemagglutinin and hemolysin in the infection process of Escherichia coli sorbitol-negative in vitro. This study was performed using 23 isolates of Escherichia coli sorbitol-negative from quails, 52.2% (12 of 23 isolates) had hemagglutinin, while 34.8% (8 out of 23 isolates) had hemolysin. Isolates with hemagglutinin were more attached to human buccal epithelial cells than isolates without hemagglutinin (P <0.05). Isolates with hemolysin were less phagocyted by macrophages compared to isolates which without hemolysin (P <0.05). Escherichia coli sorbitol-negative isolates from quails that have hemagglutinin and hemolysin are pathogenic isolates that possess the potential to cause colibasilosis and transmission between quails and other birds.

Uji pigmen dan deteksi kapsul polisakarida pada Staphylococcus aureus isolat asal broiler PIGMENT TEST AND DETECTION OF POLISAKARIDA CAPSULES ON Staphylococcus aureus ISOLAT BROILER Khusnan Khusnan; Dwi Kusmanto
Jurnal Veteriner Vol 20 No 3 (2019)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Staphylococcus aureus is a pathogenic bacterium causing disease in humans and animals. In broilers it cause septicemia, tendosinovitis, dermatitis, endocarditis, wound infections and arthritis and bumblefoot. The ability of Staphylococcus aureus to cause disease depends on the virulence factors they bear. The purpose of this research is to investigate the distribution of pigment production type and the existence of genes of polysaccharide capsule phenotype and genotype as determinant factor of virulence of bacteria on 15 isolates of Staphylococcus aureus from broiler. Pigment production test showed that 86.7% of isolates producing yellow pigment and 13.3% isolates produce orange pigment. The detection of polysaccharide capsules was phenotypically performed with hydrophobicity test with serum soft agar medium (SSA) showed 53.3% isolate grow compact and 46,7% isolate grown difuse and hydrophobicity test by salt aggregation test method (SAT) showed 66,7% hydrophil and 33.3% are hydrophobic. Genotype detection of polysaccharide capsule genes by polymerase chain reaction (PCR) showed 66.7% detected cap5 (amplicon 361bp) and 33.3% detected cap8 (173bp ampliole). The type of pigment production and the presence of polysaccharide capsules are some of the virulent factors in Staphylococcus aureus.

Staphylococcus aureus Penghasil Pigmen Kuning yang Diisolasi dari Kejadian Bumblefoot pada Broiler Lebih Patogen Dibanding Penghasil Pigmen Putih (STAPHYLOCOCCUS AUREUS PRODUCING YELLOW PIGMENT ISOLATED FROM BUMBLEFOOT CASE IN BROILER CHICKENS IS MORE PAT Khusnan .; Wahyu Prihtiyantoro; Mitra Slipranata
Jurnal Veteriner Vol 15 No 4 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (119.768 KB)

A study to evaluate the correlation between the pigments produced by Staphylococcus aureusisolateswith their adhesion capability on epithelial cells and phagocytes activity of macrophages in vitro.Ten S.aureus isolatesfrom bumble foot and arthritis casesin broiler were used in this study. The adhesion assaywas performed using epithelial cell derived from human buccal epithelial cells whereas phagocytic activitywas conducted using mouse peritoneal macrophage cells.The results showed that S. aureus isolates frombroiler produced both yellow and white pigments. Staphylococcus aureus producing white pigment adheredto human buccal epithelial cells with the density of about 24,25 bacteria/cell, which were higher than thoseof Staphylococcus aureus producing yellow pigmentabout 23,78 bacteria/cell. Staphylococcus aureusproducing white pigment was more phagocytosed by macrophages than S. aureusproducing yellow pigment,with an average phagocytic activiry of19,86 bacteria/cellas compared to15,96of bacteria/cell, respectively.

Distribusi Gen Enterotoksin Staphylococcus aureus dari Susu Segar dan Pangan Asal Hewan Siti Isrina Oktavia Salasia; Khusnan -; Sugiyono -
Jurnal Veteriner Vol 10 No 3 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (150.453 KB)

Staphylococcus aureus is a potential pathogen causing disease in human and animals due to severalvirulence factors. Staphylococcal enterotoxins which is responsible for foodborne disease is considered tobe one of the important virulence factor for the bacteria. The research was conducted to identify variousenterotoxin genes of S. aureus. Twenty three S. aureus isolates from milk cows (12 isolates) and foodanimal products (11 isolates) were used to study various enterotoxins genes (sea, seb, sec, sed, see, seg, seh,sei, and sej). The enterotoxins genes of S. aureus were investigated by using polymerase chain reaction(PCR) with specific primers. There were 3 isolates (13.04%) negative for staphylococcal enterotoxin genes.Twenty isolates (86.96%) harboured for one or more staphylococcal enterotoxin genes such as: sec (6 isolates/26.09%), see, seh for 1 isolate (4.35%), combination of 2 genes se (b,i), se (c,g), se (g,i) for 1 isolate of each(4.35%), se(c,e) for 2 isolates (8.70%), se(b,c) for 4 isolates (17.39%). Staphylococcal enterotoxin could bedetected in 3 combination genes of se(b,c,i), se(c,e,i), se(c,g,i) for 1 isolate of each (4.35%).

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