Identifikasi Isolat Khamir Berpotensi sebagai Agens Antagonis dan Uji Produksi Toksin Hemolisin

Identifikasi khamir dapat dilakukan secara konvensional maupun molekuler. Identifikasi secara konvensional membutuhkan waktu yang lama dan interpretasi hasilnya seringkali bersifat subyektif. Sementara identifikasi khamir dengan metode molekuler dapat memberikan hasil yang lebih akurat dan cepat. Khamir yang berperan sebagai agens antagonis harus aman terhadap organisme nontarget agar dapat diaplikasikan di lapangan. Penelitian ini bertujuan untuk mengidentifikasi isolat-isolat khamir berpotensi antagonis dengan metode molekuler dan mengetahui kemampuan khamir dalam menghasilkan hemolisin sebagai salah satu indikator potensi resiko terhadap mamalia. Identifikasi dan pengujian kemampuan khamir dalam menghasilkan hemolisin dilakukan pada 15 isolat khamir berpotensi antagonis terhadap patogen antraknosa cabai (Colletotrichum acutatum). Identifikasi khamir dilakukan secara molekuler dengan PCR menggunakan primer ITS1 dan ITS4. Penyediaan khamir menggunakan mediaYeast Malt Extract Broth (YMB) dan Potato Dextrose Agar (PDA). Pengujian kemampuan khamir dalam menghasilkan hemolisin menggunakan media blood agar base (Oxoid CM55) ditambah darah domba 5%. Hasil identifikasi menunjukkan bahwa isolat khamir dapat teramplifikasi dengan primer ITS1 dan ITS4 dengan ukuran fragmen produk antara 500-800 pb. Hasil analisis sekuensing didapatkan 6 spesies khamir yaitu Candida tropicalis, Rhodotorula minuta, Aureobasidium pullulans, Pseudozyma hubeiensis, Pseudozyma aphidis, dan Pseudozyma shanxiensis. Uji kemampuan khamir dalam menghasilkan hemolisin menunjukkan bahwa seluruh khamir yang diuji tidak menghasilkan toksin hemolisin sehingga diduga isolat-isolat tersebut tidak patogenik terhadap manusia.

Possibilities of practical application of different culture mediums for laboratory diagnostic of diphtheria

The purpose of the work is to evaluate the cultural and morphological properties of colonies of clinically significant corynebacteria on culture mediums for the isolation of corynebacteria. The study used 9 culture mediums for the isolation of corynebacteria: a culture medium for the isolation of corynebacteria (Corynebacagar); Tellurite-containing blood agars on base - Culture medium № 1 GRM, Culture agar for the cultivation of microorganisms (GRM agar), Culture medium for determining the sensitivity of microorganisms to antibacterial preparations - AGV, culture agar for the cultivation of dry microorganisms (SPA), Clauberg medium II, Hoyle Medium agar (Oxoid), Blood agar base (Conda), Columbia Agar Base (Conda). The work used 7 test strains of microorganisms from the State collections of pathogenic microorganisms - C. diphtheriae biovars gravis, mitis, intermedius, belfanti and subspecies lausannense, C. ulcerans and C.pseudotuberculosis. Studies were carried out in accordance with MUK 4.2.3065-13 «Laboratory diagnosis of diphtheria infection». We describe culture-morphological properties of strains on all tested culture mediums the isolation of corynebacteria after 24 and 48 hours of incubation. Analysis of the results on the growth properties of culture mediums showed that all culture mediums had high sensitivity - from dilution 10-7 for all test strains. Colonies of corynebacteria were visually detected on culture mediums after 19-20 hours of cultivation. When cultivating a suspension of corynebacteria from breeding 10-6 on culture mediums, the number of colonies ranged from 95±5 to 120±10. Conclusion. All culture mediums had differential diagnostic properties that ensure the growth of corynebacteria after the day of incubation.

Isolation of Aeromonas hydrophila in a case of wound infection in cattle in Nigeria

A three and a half year old N'dama cow weighing about 150kg was rescued from a steel jaw trap during grazing the previous day and presented at the Teaching and Research Farm Federal University of Agriculture, Abeokuta, Nigeria with a fresh, deep wound. The animal was treated but there were no satisfactory response to Penicillin/streptomycin intramuscular injection at 10,0001U/kg for seven days and also i/m 20% Oxytetracycline injection at 20mg/kg repeated after 48 hours. A loopful of pus samples aseptically taken from the wound were streaked on blood agar base enriched with 7% horse blood (Oxoid, UK) using the quadrant streaking method and incubated aerobically at 37°C for 24-48 hours. Aeromonas hydrophila was isolated which was resistant to the commonly used antibiotics on the farm. Following the results obtained from the sensitivity test, the animal was placed on Enrofloxacin for seven days, the swelling regressed, no pus was expressed from the wound site and no pain was elicited from the joint on application of pressure by the 8th day Aeromonas hydrophila is pathogenic to man; hence care should be taken in handling of animal traps.

Efecto in vitro del extracto de Solanum sessiliflorum “cocona” sobre el crecimiento de Heli­cobacter pylori

El objetivo del presente trabajo fue determinar y comparar el efecto in vitro del extracto de Solanum sessiliflorum “cocona” sobre Helicobacter pylori frente a los antibióticos: amoxicilina, tetraciclina, claritromicina y amikacina. La susceptibilidad para estos antibióticos fue determinada por el método de difusión de discos y el efecto del extracto de cocona sobre Helicobacter pylori por el método de placa vertida. Las cepas de Helicobacter pylori fueron obtenidas de biopsias gástricas y los medios utilizados fueron Columbia blood agar base y Mueller Hinton suplementado con 5% de sangre de carnero y, para la reactivación de las cepas de Helicobacter pylori, un suplemento específico Dent (Vancomicina 5 mg, trimetropina 2,5 mg, cefsulodina 2,5 mg y anfotericina B 2,5 mg); el ambiente microaerofílico se consiguió empleando OXOID Campy Gen. Los resultados confirman el efecto inhibitorio in vitro del extracto de cocona sobre Helicobacter pylori, siendo superior al de los antibióticos usados.

Reliability of Selective Media for Recovery of Staphylococci from Cheese

The reliability of 12 selective media for recovery of four different strains of Staphylococcus aureus inoculated at manufacture into three cheese types was determined. Selective medium and time of ripening had a highly significant effect (p < 0.001) on reliability of the staphylococcal count. In addition, highly significant interaction effects were observed. The most reliable medium in the overall analysis was mannitol salt agar. However, this medium was not equally reliable at all times during ripening, and use of both mannitol salt agar and Staphylococcus medium no. 110 is recommended. The tellurite- and azide-based selective media were generally unsatisfactory, however tellurite glycine agar, Vogel Johnson (VJ) agar, and azide blood agar base were totally unreliable. In general, the salt-based selective media were most reliable. This applied also to the egg yolk media that use salt as the selective agent. Salt egg yolk agar and Colbeck's egg yolk medium generally gave higher recoveries of S. aureus than did Baird-Parker medium, Crisley et al. tellurite polymyxin egg yolk agar, and Hopton egg yolk azide agar, except in the unripened cheeses. The debilitating effect of cheese ripening on the staphylococcal cells was not eliminated by the egg yolk tellurite and azide media.

Bacillus subtilis GSY 1057 Assay for Aflatoxin B1 Activation by Rainbow Trout (Salmo gairdneri)

A rapid and sensitive microbial assay was developed to detect lethal products of anatoxin B1 metabolism by rainbow trout (Salmon gairdneri) Mt. Shasta strain. Bacillus subtilis GSY 1057 (hisA1, uvr-1, metB4), a DNA repair deficient strain, was incubated for 20 min in the 20,000 × g supernate from trout liver homogenates which had been preincubated for 10 min with various levels of aflatoxin B1. Serial dilutions of the incubation mixture were plated in triplicate on tryptose blood agar base plates and colonies were counted after 12 hr at 37°C. One μmole aflatoxin B1 in 3.2 ml incubation mixture reduced viability 60°.