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

Karakterisasi Feno-Genotipik Kemampuan Hemolisa Isolat Staphylococcus aureus Asal Susu Kambing Mastitis dan Daging Ayam Segar

Abstract  Staphylococcus aureus (S. aureus) is a pathogenic bacterium with many virulence factors, such as toxins, immunomodulatory factors, and exoenzymes. Hemolysin is one of the essential toxins in the pathogenesis of S. aureus, with the action of lysing erythrocytes and other cells. This study aimed to determine the phenotypic and genotypic characteristics of S. aureus isolates from animal origin. Eight S. aureus isolates from goat mastitis and raw chicken meat phenotypically evaluated the ability to hemolyze sheep blood. At the same time, genotypic were determinated by detecting the hla and hlb genes. The results showed that most 87.5% (7/8) of S. aureus isolates had β-hemolysis phenotypic. PCR determination showed that all (8/8) isolates were positive for the α-toxin (hla) encoding gene, while half (4/8) of the isolates were positive for the β-toxin (hlb) encoding gene. The use of 5% defibrinated sheep blood agar plates is recommended to determine the hemolytic character of S. aureus tested. Keywords: Character; Hemolysis; S. aureus   Abstrak  Staphylococcus aureus (S. aureus) merupakan bakteri patogen yang memiliki banyak faktor virulensi seperti toksin, faktor imunomodulator, dan eksoenzim. Hemolisin merupakan salah satu toksin penting dalam patogenesis S. aureus, dengan aksi kerja melisiskan eritrosit dan sel lainnya. Penelitian ini bertujuan untuk mengetahui karakter hemolisa isolat S. aureus asal hewan secara fenotipik dan genotipik. Delapan isolat S. aureus asal susu kambing mastitis dan daging ayam segar dievaluasi secara fenotipik untuk melihat kemampuan menghemolisa darah domba, sedangkan determinasi genotipik dengan deteksi gen hla dan gen hlb. Hasil penelitian menunjukkan mayoritas 87.5% (7/8) isolat S. aureus memiliki fenotipik β-hemolisa. Determinasi PCR menunjukkan 100% (8/8) isolat positif gen penyandi α-toksin (hla), sedangkan 50% (4/8) isolat positif gen penyandi β-toksin (hlb). Penggunaan 5% defibrinated pelat agar darah domba lebih direkomendasikan untuk melihat karakter hemolisa S. aureus yang diuji. Kata kunci: Hemolisa; Karakter; S. aureu