Addition of insoluble fiber to isolation media allows for increased metabolite diversity of lab-cultivable microbes derived from zebrafish gut samples

There is a gap in measured microbial diversity when comparing genomic sequencing techniques versus cultivation from environmental samples in a laboratory setting. Standardized methods in artificial environments may not recapitulate the environmental conditions that native microbes require for optimal growth. For example, the intestinal tract houses microbes at various pH values as well as minimal oxygen and light environments. These microbes are also exposed to an atypical source of carbon: dietary fiber compacted in fecal matter. To investigate how the addition of insoluble fiber to isolation media could affect the cultivation of microbes from zebrafish intestines, an isolate library was built and analyzed using the bioinformatics pipeline IDBac. The addition of fiber led to an increase in bacterial growth and encouraged the growth of species from several phyla. Furthermore, fiber addition altered the metabolism of the cultivated gut-derived microbes and induced the production of unique metabolites that were not produced when microbes were otherwise grown on standard isolation media. Addition of this inexpensive carbon source to media supported the cultivation of a diverse community whose specialized metabolite production may more closely replicate their metabolite production in vivo.

Diversity of Culturable Actinomycetes from Deepsea Floor of Makassar Strait, Indonesia

With regard to collaboration research called Widya Nusantara Exploration (EWIN) in May-June 2013 and November 2014, a study on isolation of actinomycetes from sediments of Makassar Strait have been conducted. Actinomycetes is one of microbe which has an excellent track record in producing antimikrob and other active substances. But due to terrestrial actinomycetes has been widely explored, then recently researchers began focusing on wide variety of extreme environments, such as marine environment, to screening aktinomisetes in producing new secondary metabolites. A total of 36 strains of actinomycetes were isolated from 10 samples obtained from deepsea floor in Makassar Strait, Indonesia, Direct Dillution Method were best used to isolate the actinomycetes compare to Sodium Dodecyl Sulfida – Yeast Extract Method (SDS-YE Method) and Rehidration Centrifugation Method (RC Method). NBRC-802 media and Actinomycetes Isolation Agar(AIA)(Himedia)media were used as the isolation media. All the isolates were identified by morphological characteristic and by analysis of 16S rRNA gene sequence. Actinomycetes isolated from deepsea floor of Makassar Strait have been dominated by Micromonospora (58%), Verrucosispora (14%)Streptomyces (8%) and Luteipulveratus (5%), however genus Nocardiopsis, Micrococcus, Gordonia, Kytococcus, and Arthrobacter were not dominant (3%). Station 25 in 1.547 m depth was the most abundant of actinomycetes, 18 strains and dominated by the genus Micromonospora which is isolated using Direct Dillution Method and both NBRC 802 or AIA media.

IDENTIFIKASI SALMONELLA TYPHI PADA PENDERITA DEMAM TIFOID DI PUSKESMAS MALILI

The success of epidemiological survey of Salmonella is associated with procedures adopted to differentiate the genotypes of Salmonella typhi. The research aimed identification of Salmonella typhi from the blood culture of typhoid fever suspect.The research was conducted in the Microbiology Laboratory, Faculty of Medicine, Hasanuddin University. Samples were taken from Public health centers (PHC) in Malili. This was an analytic quantitative research with the cross sectional approach. Samples were collected from the blood of the patients with suspected positive typhoid fever suspects. The samples were cultured to obtain S.typhi isolates finally the data were analysed.The research result indicates that in the culture examination, the isolation result of the Salmonella typhi from the blood samples of 98 patients, its obtained 20 (20.4%) positive typhoid fever samples. This show that successfull identification of Salmonella typhi still lower. This is depend of several factor such as using antibiotic before take of blood, limited used of isolation media, total of less bacteria in blood, less volume of blood and time to take of blood not exactly.Keywords : Salmonella typhi,isolation, typhoid fever

Pengaruh Nutrisi Dan Suhu Terhadap Selektivitas Potensi Antibakteri Dari Bakteri Yang Berasosiasi Dengan Spons

The Influence of nutrient and temperature to the antibacterial selectivity of Sponge Associated-Bacteria Production of pharmacological activity by marine microorganism is strongly influenced by nutrition and environmental conditions. In this study would discuss about the influence of several type of media to the production of antibacterial agent by sponge-. associated microorganisms. About 3 sponges tissue Theonella sp, Callispongia sp. and Lithistide sp. collected from Seribu Island will be used for the host of associated microorganism. Agar medium used for isolation were M1 that contained amylum, yeast extract and peptone, M2 (10% marine broth media) contained yeast extract and peptone, M3 only sea water without adding any nutrients. Beside the nutrient variation, heat sock treatment at 50oC toward the sponge solution also apply to this study. The bacterial isolation data indicated that bacterial density in (CFU/100µL) of Theonella sp, Callispongia sp. and Lithistide sp. were minimum when spreading in M3 medium with heat sock treatment. This data showed that limiting in nutrient and heating could increasing bacterial selectivity. The antibacterial activity capability of bacterial strains isolated using M1, M2 and M3 respectively in range were 81,8-90,9%; 50-87,5% and; 66,7 -100%. This results showed that less nutrient of media will rise the number of antibacterial activity strains,and decreasing of bacterial density. This study reported that the minimum nutrient of isolation media and heat shock treatment could be used for selecting the antibacterial strains of sponge associated bacteria. Keywords : isolation media, antibacterial, nutrient, microbial symbiont AbstrakAktifitas farmakologi yang dihasilkan oleh mikroorganisme laut sangat dipengaruhi oleh nutrisi dan kondisi lingkungan. Hal tersebut mendorong untuk digali lebih dalam tentang aspek-aspek yang mempengaruhi seleksi mikroba potensial pada spons. Metode isolasi mikroba dari jaringan spons menjadi kunci dalam menguak potensi mikroba simbionnya. Dalam penelitian ini akan membahas pengaruh berbagai media isolasi bakteri yang berasosiasi dengan spons asal Kep. Seribu. Terhadap 3 specimen spons Theonella sp., Callispongia sp. dan Lithistide sp. dilakukan isolasi bakteri dengan metode direct sampling menggunakan media M1, M2 dan M3. Media M1 mengandung nutrisi antara lain amilum, ekstrak khamir dan pepton, sedangkan media M2 mengandung sumber nutrisi ekstrak khamir dan pepton dan M3 hanya media agar dan air laut. Selain variasi nutrient dalam media, perlakuan pemanasan pada suhu 50oC juga akan dilakukan terhadap larutan sampel spons sebelum dilakukan penyebaran pada media isolasi. Hasil isolasi bakteri yang diisolasi spons Theonella sp, Callispongia sp. dan Lithistide sp. menunjukkan bahwa kepadatan minimum diperoleh dengan menggunakan media M3 dengan perlakuan pemanansan. Dari data isolasi bakteri menunjukkan bahwa selain kandungan nutrient yang minimum, perlakuan pemanasan akan menurunkan kepadatan jumlah bakteri yang tumbuh, sehingga pemanasan merupakan salah satu cara dalam seleksi isolasi bakteri yang berasosiasi dengan spons. Hasil analisis aktivitas antibakteri menunjukkan bahwa persentase strain-strain bakteri yang aktif terhadap antimikroba Vibrio eltor, Eschericia coli dan Bacillus subtilis dengan variasi media M1 berkisar antara 81,8-90,9%; M2 berkisar 50-87,5% dan M3 berkisar 66,7-100%. Dari data tersebut disimpulkan bahwa semakin sedikitnya nutrisi media isolasi maka semakin tingginya mikroba-mikroba potensial penghasil antibiotik. Media M3 merupakan media yang selektif untuk isolasi mikroba potensial dari spons, terbukti dengan tingginya prosentase bakteri yang aktif dan berkurangnya jumlah koloni yang tumbuh.Kata kunci : media isolasi, antibakteri, nutrisi, mikroba simbion

Larvicidal potentials of Metarhizium anisopliae and Paecilomyces sp on Culex quinquefasciatus (Say) (Diptera: Culicidae)

Biological control potential of Metarhizium anisopliae and Paecilomyces species against Culex quinquefasciatus was evaluated. M. anisopliae and Paecilomyces spp was isolated from soil using soil suspension method with selective isolation media. Bioassay was made to determine their efficacy against 4th instar larvae of C. quinquefasciatus. Three different concentrations; 1x106, 1x107 and 1x108conidia/ml were made and tested. Results showed that, mortality increased as the period of exposure increased and also increased as the conidia concentration increased. The mortality recorded in lowest dose of 106 conidia/ml was 60% and 50% in M. anisopliae and Paecilomyces spp respectively, and again 80% and 70%mortality was recorded at dose of 107 conidia/ml. whereas highest dose level of 108conidia/ml caused high mortality of up to 90% and 80% respectively. The lethal concentration causing 50% mortality (LC50) of 4th instars larvae of C. quinquefasciatus was also varied according to concentration of spores and duration of exposure. The result showed that LC50 values of M. anisopliae isolate were 3.8×108, 2.5 x 106, 2.0 x 105 and 1.7x107conidia/ml after 24, 48, 72 and 96 Hours exposure respectively. Similarly, LC50 values Paecilomyces Spp isolate were 5.3×108, 3.8 x 107, 2.0 x 106 and 2.3 x 107conidia/ml after 24, 48, 72 and 96 Hours exposure respectively. the These results indicated that M. anisopliae and Paecilomyces Spp isolated is pathogenic to immature stage of C. quinquefasciatu and could be suggested for development as a biological control for mosquitos’ management.