Analisis Keragaman Genetik Streptococcus agalactiae Penyebab Mastitis Subklinis Pada Sapi Perah Menggunakan pulsed field gel electrophoresis (PFGE) = Genetic Analysis of Streptococcus agalactiae Caused Subclinical ...

Streptococcus agalactiae atau Streptokokus grup B (SGB) adalah salah satu bakteri utama penyebab mastitis subklinis pada sapi perch dan merupakan parasit obligat pada ambing. Karakterisasi S.agalactiae biasanya dilakukan secara konvensional menggunakan metode serotyping. Meslci metode ini sering digunakan namun masih mempunyai kelemahan apalagi masih adanya isolat S.agalactiae yang belum dapat dimasukkan ke dalam serotipe yang ada (nontypeable/NT), oleh karena itu pendekatan bare dengan metode genotyping digunakan. Tujuan dari penelitian ini adalah melihat profil DNA menggunakan PFGE dan kekerabatan isolat dari masing-masing serotipe maupun masing-masing daerah. Penentuan serotipe S. agalactiae dilakukan dengan metode serotyping menggunakan antiserum spesifik terhadap 9 serotipe S. agalactiae dengan uji imunodifusi/ agar gel presipitasi (AGP). Analisa genotipe S. agalactiae dilakukan menggunakan macro restriction fragment length polymorphism (MLFP)/ metode schizotyping menggunakan Pulsed-field gel electrophoresis (PFGE). Hasil dari penelitian ini adalah genotipe dari S. agalactaie dengan enzim restriksi Smal dihasilkan potongan-potongan pita Deoxyribo Nucleic Acid (DNA) yang jelas. Ada 3 isolat S. agalactiae yang tidak dapat dipotong oleh ensim restriksi Smal. Analisa DNA genom dari 21 isolat S.agalactiae dihasilakn 15 profil DNA. Kata kunci: mastitis subklinis, Streptococcus agalactise, PFGE Streptococcus agalactiae is one of the main agents responsible for subclinical mastitis in dairy cattle and as an obligate parasite in udder. Characterization of S. agalactiae usually done by conventional serotyping method. Although this method is often used but actually the method had lack of a sufficiently discriminatory typing system and the existence of nontypeable (NT), therefore a new approach by genotyping used. The aims of this research were to know the profile of DNA S. agalactiae and the genetic relationship between isolates. Determination of serotype of group S. agalactiae was done by serotyping method using specific antiserum to S.agalactiae in immunodifusion assay/ Agar Gel Precipitation Test (AGPT). The genotypic of the bacteria was done by macro restriction fragment length polymorphism (MFLP)/ schizotyping method using Pulsed-field gel electrophoresis (PFGE). Results of the present study showed that genotypic of S. agalactiae with Smal restriction enzyme produced Deoxyribo Nucleic Acid (DNA) discrete fragments . Threre were 3 isolates of S. agalactiae that could not be restricted by Smal enzyme. Genomic DNA analysis of 21 isolates showed 15 different DNA profiles. Key words: subclinical mastitis, Streptococcus agalactise, PFG

Risk Factor Identification and Incidence Rate Measurement on One High-Risk Farm as Brucellosis Precautions in Metro City Lampung

Since 2011, Lampung is one of the Brucella-free areas, but April 2021 reported abortion case in a cow farm     at Metro city Lampung Province. The purpose of this study was to identify risk factors and measure incidence levels as vigilance re-emerging Brucellosis. Sample of cow serum, taken in a census from whole cattle cow in a farm at risk, as much as 2 times with a distance about 1 month. In the first census, 703 serum samples were taken from the stables and 25 samples from the adjacent. Whole sample tested to Brucella spp with rose bengal test (RBT) and positive result confirmed with complement fixation test (CFT). The second census was conducted on samples without positive CFT results in the first census. The positive RBT at second sensus were followed by CFT and c-Elisa tests. Incidence rate of brucellosis in high-risk housing in Banjarsari Village, North Metro District, Metro City 1,67% or 2 brucellosis per 100 cows a month. Brucella spp. seropositivity was significantly related to the type of species and sex of the animal; animals with Ongole breed 4.2 times and female animals 4.5 times have a higher level of exposure compared to non-Ongole breed animals and males

Uji In Vitro Efektivitas Ekstrak Biji Jintan Hitam (Nigella Sativa L.) terhadap Pertumbuhan Microsporum gypseum Penyebab Dermatitis pada Anjing

Biji jintan hitam (Nigella sativa L.) mengandung senyawa aktif seperti thymoquinone, carvacrol, dan thymol yang berperan sebagai antijamur. Penelitian ini bertujuan untuk mengetahui kemampuan ekstrak biji jintan hitam dalam menghambat pertumbuhan jamur Microsporum gypseum yang merupakan salah satu agen penyebab dermatitis pada anjing. Kemampuan ekstrak biji jintan hitam dalam menghambat pertumbuhan jamur Mircrosporum gypseum diuji dengan metode difusi agar dengan teknik sumuran (Agar Well Diffusion). Rancangan penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan 7 perlakuan konsentrasi ekstrak biji jintan hitam terhadap pertumbuhan jamur Microsporum gypseum yaitu 10%, 25%, 50%, 75%, 100%, flukonazol digunakan sebagai kontrol positif dan phospat buffer saline (PBS) kontrol negatif. Hasil penelitian menunjukkan bahwa ekstrak biji jintan hitam dengan konsentrasi mulai dari 25%, 50%, 75% dan 100% dapat menghambat pertumbuhan jamur dengan rerata lebar zona hambat secara berurutan ± 3,67 mm., ± 2,84 mm., ± 3,67 mm., dan ± 4,00 mm. Ekstrak biji jintan hitam mampu menghambat pertumbuhan Microsporum gypseum.  Perlu penelitian lebih lanjut untuk mendapatkan hasil optimal sehingga berpeluang sebagai anti fungal pada penyakit dermatitis pada anjing

Karakterisasi Hemaglutinin Streptococcus agalactiae dan Staphylococcus aureus Penyebab Mastitis Subklinis Pada Sapi Perah = Characterization of Haemagglutinin of Streptococcus agalactiae and Staphylococcus aureus on ...

Streptococcus agalactiae dan Staphylococcus aureus merupakan dua bakteri utama penyebab mastitis subklinis pada sapi perah di Indonesia. Pada mastitis subklinis kemampuan adesi mempunyai peran lebih penting dari pada kemampuan invasi. Hemaglutinin merupakan faktor yang berperan dalam proses adesi, sebagai langkah awal kolonisasi bakteri pada permukaan set epitel ambing. Tujuan dari penelitian ini adalah mengisolasi dan mengkarakterisasi hemaglutinin dari S. agalactiae dan S. aureu. Penentuan hemaglutinin S. agalactiae dan S. aureus dengan uji hemaglutinasi menggunakan eritrosit sapi perah 1%. Isolasi hemaglutinin dilakukan dengan afinitas kromatogarfi dan analisis dengan Sodium Dodecyl Sulphate-Polyacrilamide Gel Electrophoresis (SDS-PAGE). Hasil penelitian diketahui bahwa hemaglutinin S. agalactiae dan S. aureus dapat ditentukan dengan SDS-PAGE dengan berat molekul 28 kDa pada S. agalactiae dan 27 kDa pada S. aureus. Kata kunci : Streptococcus agalactiae, Staphylococcus aureus, mastitis subklinis, hemaglutinin Streptococcus agalactiae dan Staphylococcus aureus are two main bacteria that caused subclinincal mastitis in dairy cows in Indonesia. In the pathogenesis of subclinical mastitis, the adhesion process is more important as the initiation step of bacterial colonization on the mamary cell surface. Haemagglutinin play a role as adhesins in mediating the adherence of this bacteria. The aim of this research was to isolate and to characterize of haemagglutinin of S. agalactiae and S. aureus. To determine of S. agalactiae and S. aureus that had haemagglutinin were done by haemagglutination test with 1% of erythrocyte of dairy cattle. Isolation and characterization of haemagglutin were done by using affinitas chromatography technique and characterized by Sodium Dodecyl Sulphate-Polyacrilamide Gel Electrophoresis (SOS-PAGE). The results of the present study showed that haemagglutinin of S. agalactiae and S. aureus could be separated by SDS-PAGE with molecular weight of 28 kDa for S. aureus and 27 kDa for S.agalactiae. Key words: Streptococcus agalactiae, Staphylococcus aureus, subclinical mastits, hamagglutini

Uji Kepekaan Avibacterium paragallinarum Terhadap Antibiotik yang Berbeda

Avibacterium paragallinarum is the aetiology of Infectious coryza (snot), one of acute upper respiratory tract diseases, which causes financial loss due to the decrease of egg production in layer. This is Gram negative bacteria which have three serotypes, known as serotype A, B, and C. The aim of this research was to know the sensitivity of Avibacterium paragallinarum to different kinds of antibiotics that are commonly used. This research used Avibacterium paragallinarum culture derived from layer which showed the symptoms of snot. Sensitivity test applied erythromycin, enrofloxacin, gentamycin, amoxicillin and clavulanic acid, and sulfamethoxazol. Results found the sensitivity of Avibacterium paragallinarum to combination of amoxicillin and clavulanic acid was 100%, to erythromycin 50% resistant and 50% intermediate, to enrofloxacin 75% resistant and 25% intermediate, to gentamycin 75% sensitive and 25% resistant, and to sulfamethoxazol 100% resistant

Isolasi, Identifikasi, dan Serotyping Avibacterium paragallinarum dari Ayam Petelur Komersial yang Menunjukkan Gejala Snot

Infectious coryza (snot) is one of acute respiratory disease in breeders, layers, and broilers caused by Avibacterium paragallinarum (Av. paragallinarum). This disease is very harmful because of its cause decrease in egg production and high morbidity. Snot incident in Indonesia still is reported. Vaccination is one of the best preventive measures, but reports about Av. paragallinarum serotype at the field so lack so that the correspondence between serotype Av. paragallinarum in the field to those used for vaccination is unknown. Av. paragallinarum has strains with different antigenicity and until now known three serotypes there are serotypes A, B and C. Serotypes A and C are pathogenic then serotypes B. The purpose of this study was to isolate, identify and to determine the serotype of Av. paragallinarum from the commercial layer that showing symptoms of snot. Samples were taken from layer which showed symptoms of snot (nasal foul smelling exudate, infra-orbital and wattle swelling, conjunctivitis, and snoring) from some layer farms. Samples were cultured on chocolate agar and then incubated in a candle jar at 37 ᴼC for 18-24 hours. Bacteria colony and cell morphology were observed and performed biochemical tests (catalase, oxidase, urease, indole, and fermentation of carbohydrates) in suspected colonies. Serotypes test was conducted using plate agglutination test (PAT). This study revealed 4 isolates Av. paragallinarum with 2 isolates are serotype B and 2 other are serotype C

Combination of Wild ginger, Honey, and Probiotics as Growth Prometer Candidates : An in Vitro Study

Growth Promoter Antibiotics are used to prevent disease and promote growth and production in poultry. Repeated admnistration of feed can have a microorganic resistance effect, accumulation of antibiotic residues in animal and environmental products and imbalance of normal microflora in the intestine. The antibacterial and carbohydrate content of some natural ingredients can be potential as a replacement candidate for AGP. This study aims to determine the role of a combination of wild ginger, honey, and probiotics (Bacillus subtilis and Lactobacillus acidophilus) as AGP candiate in Vitro. The antibacterial activity of the combination of wild ginger and honey against pathogens (E. coli) and their use against probiotics was tested by disk diffusion method, while the calculation of optical density values to determine the minimum inhibitory concentration and minimum bactericidal concentration was carried out on E. coli. The ability of inhibition of probiotics against pathogens is also done by the disk diffusion method. The disk diffusion test results showed the best combination was extract of 25% wild ginger aquades + 100% Lombok honey with inhibition zone diameter (8,53 ±, 03). Optical density values indicate this combination is able to inhibit and kill E. coli (DO 0,00 ±, 002) and support B. subtilis (DO 0,18 ±, 002) and L. acidophilus (DO 0.25 ±, 005) significantly better than positive control. MIC value of wild ginger aquades extract and honey combination against E. coli is wild ginger aquades extract 3.13% + Lombok honey 25%, while MBC value is wild ginger aquades extract 6,25% + Lombok honey 25%. The combination of B. subtilis and L. acidophilus showed the largest inhibitory zone diameter against E. coli (7,30 ±, 02 mm) compared to individual colonies. The combination of wild ginger and honey, in addition to inhibiting also able to kill pathogens and support the growth of probiotics, so this formula can be used as one of the replacement candidates for AGP. 

MP-16 Characterization of Avibacterium paragallinarum Caused Infectious coryza/Snot: Satellite Colony Phenomenon

Infectious coryza (IC) is an acute upper respiratory disease of poultry that can appear in all ages. Some of clinical signs that are commonly seen in IC are rhinitis, facial swelling or edema, lacrimation, anorexia, and retarded growth in young poultry [1.2.3]. The disease can be found worldwide, especially in tropical countries [4]. Infectious coryza is very important in the chicken farm industry in developed and developing countries, including Indonesia [5]. The large economic losses due to IC such as increased number of culling, decreased egg production (10-40%), decreased body weight, stunting growth, and some mortality (2-10%) [4].Avibacterium paragallinarum which was previously classified as Haemophilus paragallinarum is a causative agent of infectious coryza in laying and broiler chickens, quail, pearl chicken, turkey, and peacocks [4,6,7,8]. The bacteria is commensal in the mucous membrane of the upper respiratory system, is sensitive to preservation and does not last long outside the host body [8]. Factors X and V are needed for the growth of several types of A. paragallinarum. According to in vitro growth requirements, A. paragallinarum can be either nicotinamide adenine dinucleotide (NAD) independent or NAD-dependent. The reduced form of NAD (NADH; 1.56-25 µg/ml) and the oxidized form (20-100 µg/ml) is required for in vitro growth in most isolates A. paragallinarum that show satellitic colony on a medium [9]. The description of the need for V factor of field isolates A. paragalinarum has been few reported. The aim of this research is to find out the phenomenon of satellite colonies from a variety of poultry isolates

EVALUASI STATUS VIRULENSI ISOLAT Bacillus anthracis ASALNUSA TENGGARA DAN PAPUA MENGGUNAKAN METODE POLYMERASE CHAIN REACTION MULTIPLEX

Penelitian ini bertujuan mengevaluasi status virulensi 22 isolat Bacillus anthracis (B. anthracis) asal Nusa Tenggara dan Papua  menggunakan metode polymerase chain reaction (PCR) multiplex dengan dua pasang primer nukleotida yang memiliki target amplifikasi gen spesifik pada kedua plasmid. Ektraksi DNA dilakukan dengan metode lisis panas. Pasangan primer PA5 dan PA8 digunakan untuk mengamplifikasi gen pagA pada pXO1, sedangkan pasangan primer 1234 F dan 1301 R mengamplifikasi gen capABC pada pXO2. Hasil reaksi PCR menghasilkan dua pita DNA berukuran sekitar 600 dan 800 bp pada 20 isolat. Namun, dua isolat lain, masing-masing hanya memiliki salah satu dari kedua ukuran pita DNA tersebut. Sebagian besar koleksi isolat asal Nusa Tenggara dan Papua (91%) masih memiliki kedua plasmid secara lengkap (pXO1+/2+) dan karena itu bersifat virulen, sedangkan dua isolat lain (9%) telah kehilangan salah satu plasmid virulennya sehingga bersifat avirulen. Disimpulkan bahwa PCR multiplex dengan dua pasang primer dengan target amplifikasi pada plasmid dapat digunakan untuk evaluasi status virulensi isolat B. anthraci

Deteksi Virus Newcastle Disease pada Burung Merpati (Columba Livia) dan Burung Tekukur (Streptopilia Chinensis) yang Menunjukkan Gejala Syaraf

Dewasa ini dilaporkan banyak kasus pada burung merpati (Columba Livia) dan burung tekukur (Streptopilia Chinensis) menunjukkan gejala syaraf, terutama: tortikolis, dan kepala gemetar, yang merupakan indikasi penyakit Newcastle Disease (ND). Kedua spesies burung tersebut banyak berkeliaran di lokasi farm ayam komersial untuk mencari pakan. Kondisi tersebut dapat bertindak sebagai faktor penular penyakit ND. Penelitian ini bertujuan mendeteksi virus ND pada burung merpati dan burung tekukur yang memperlihatkan gejala saraf, dengan uji serologis dan molekuler. Sampel darah diambil dari vena brakhialis, diproses untuk mendapatkan serum darah. Serum tersebut selanjutnya diuji dengan uji hemaglutinasi inhibisi (HI), untuk mendeteksi titer antibodi ND. Sampel pool otak, trakea, dan lien diekstraksi RNA-nya dan diamplifikasi dengan primer spesifik gen F virus ND. Sampel pool tersebut juga dikultur pada telur ayam berembrio specific pathogen free (TAB-SPF). Identifikasi dilakukan dengan uji hemaglutinasi (HA) dan HI dengan serum kontrol positif ND. Hasil deteksi serologis 7 sampel burung merpati menunjukkan titer antibodi ND bervariasi dari titer 25 sampai 28., sedangkan 2 sampel diperoleh seronegatif dengan titer 20. Salah satu pool gerusan organ kode M3/Sleman/2021 menunjukkan hasil RT-PCR positif. Analisis sekuens isolat virus ND tersebut termasuk NDV virulen dan dikelompokkan ke dalam genotipe VII-i. Pasase pool sampel organ tersebut masing-masing dikultur pada TAB-SPF dengan pasase sebanyak 3 kali, menunjukkan hasil negatif