BIO VAILIBILTY OF BACTEROIDES MELANINOGENICUS IN HEN EGG. Soeherwin Mangundjaja Department of Microbiology Faculty of Dentistry Universitas Indonesia

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1 BIO VAILIBILTY OF BACTEROIDES MELANINOGENICUS IN HEN EGG. Soeherwin Mangundjaja Department of Microbiology Faculty of Dentistry Universitas Indonesia Abstract The majority of oral flora are anaerobic bacteria. Gingival sulcus is the main port d' entre of oral anaerobic bacteria in invading other organs. Objectives:The objective of this research is to study the viability anaerobic bacteria Bacteroides melaninogenicus in hen eggs. Method: Subject of the research is Bacteroides melaninogenicus inoculated in yolk sac an incubated at room temperature for 1,2,3, and 24 hours. Results: showed that Bacteroides melaninigenicus can be isolated from 10 chicken's egg, while in control Bacterides melaninogenicus can not be found. Conclusion: Therefore we conclude that hen egg can be used as temporary media for anaerobic bacteria. Key words: Bacteroides melaninogenicus, hen,egg. Introduction Success in overcoming infectious diseases is relating closely to etiologic bacterial factor and current of the sensitivity test. Special for anaerobic bacteria, success in isolating the bacteria depends very much method used in order that the bacteria will not die during delivery to laboratory. To this end we can use several methods: 1. By making the laboratory personnel come to the patients place. 2. By shortening the period, in which the bacteria are not in optimum condition. 3. By using transport medium and special equipment for anaerobic bacteria. Those requirements are still difficult to be fulfilled by doctors working in rural areas, so we shall think it seriously. To overcome that problem, we will carry out initial research of Bacteroides melaninogenicus viability in hen eggs incubated for 24 hours.

2 Objective and Benefit of Research Currently with better isolation technique we find more species of anaerobic bacteria in periodontal abscess than before, and their prevalence is greater than that of anaerobic bacteria. This is new era for dentist, therefore anaerobic bacteria in dentistry shall be studied further. This research is survey with objective of: 1. Obtaining information regarding viability of Bacteroides melaninogenicus in he eggs. 2. Obtaining information regarding optimum growth of Bacteroides melaninogenicus in hen egg. 3. Obtaining information regarding period of viability of Bacteroides melaninogenicus in hen egg. Material and Methods Material 1. Wild strain of Bacteroides melaninogenicus originating from sulcus gingivalis of healthy person. 2. Brain Heart Infusion broth and Brucella blood agar media 3. Egg of non domestic chicken (10) Method To obtain pure culture of Bacteria originating from sulcus gingivalis, samples are taken from patients coming to Dental Clinic of Faculty of Dentistry, University of Indonesia. And then the samples are incubated anaerobically in anaerobic jar. A. Chart of microorganism isolation from sulcus gingivalis. Samples from sulcus gingivalis Brain Heart Infusion Broth + menadion + haemin Blood base agar Brucella Ultra Violet lamp Colonies red Bacteroides species 2

3 B. Bacterial Inoculums To obtain optimum number of bacterial inoculums, we use Colony Forming Units method of viable count of Colonies (George A. Wisteich). The resulting number of Bacteroides melaninogenicus inoculums is 3 X 10 4 /ml. C. Inoculating bacteria into hen egg a. The egg is placed with its broader end as the bottom, and air sac on its top, then its upper part is painted with concentrated Betadine solution and covered with sterile gauze, then we wait for 1 minute. b. Without removing the betadine, we drill the egg until we reach its membrane. c. Sample of bacteria amounting to 0.5 ml is injected into yolk sac. d. The hole is covered with cellophane e. The egg is incubated at room temperature. D. Control Sterile physiologic saline solution is injected into yolk sac and treated similarly with the above. E. Incubation of bacteria in yolk sac At room temperature for: 1 hour 2 hours 3 hours 24 hours Harvesting examination material from yolk sac is carried out by sucking the yolk with syringe of 5 mm, and we take approximately 2 ml of yolk. Then we culture the material according to chart of isolating microorganism from sulcus gingivalis. F. Result of Bacteria harvest from Yolk Sac Period of incubation Bacteria's growth hour Growing 2 hours Growing 3 hours Growing 24 hours No growth

4 Discussion Chicken's egg has high biological value as food, so we may conclude that all materials for Bacteroides melaninogenicus growth is contained in chicken's egg. Inoculation of Bacteroides melaninogenicus is carried out by inoculating 38 X 10 4 inoculum amounting to 0.5 ml with disposable syringe of 0.4 X G long into yolk sac in height angle direction with remaining length of needle 1 em from egg shell. The bacteria is harvested after 1 hour, 2 hours, 3 hours and 24 hours of incubating, then the bacteria is isolated and incubated anaerobically. The result of the entire incubation shows that Bacteroides melal1il1ogel1icus grows well in yolk sac. On other hand, control with similar method shows negative growth. The research also reveals that fresh egg or not been kept too long eggs show resistance to bacterial invasion. However, viability Bacteroides melaninogel1icus in yolk sac means that eggs can be made as temporary media. Relating to problem of transportation media for anaerobic bacteria from rural areas to laboratory, we suggest that chicken's egg can be used as substitute for special anaerobic cylinder. This research is still preliminary research, and further research shall be that infected material is inoculated directly to hen egg. Conclusion Temporarily we conclude that anaerobic bacteria can grow in chicken's egg in 24 hours, therefore chicken's egg can be used as transportation media for anaerobic bacteria such as Bacteroides melaninogenicus. References Abdul Rahim Hasil Perkembangan Teknik Isolasi Bateri Anaerobic dari bahan Pemeriksaan Klinik sejama tahun 1979 di Jakarta. (A New technique for anaerobic bacteria's isolation from clinical infected material in 1979 in Jakarta) 4

5 Sujudi 1979 : Suatu tinjauanterhadap infeksi anaerobik di Indonesia. Simposium Infeksi Anaerobik. Jakarta 5 Mei Slamet Djais 1979 :Suatu pemikiran melengkapi laboratorium Mikrobiologi Rumah Sakit Sumber Waras dengan bagian Anaerob. Simposium Infeksi anaerobic Jakarta 5 Mei 1979 Sklavounos et a Anaerobic bacteria in dento-alveolar abscesses. Int. J Oral Maxillo surgery 1986; 15: Cotteril et al 1927: A Nutrient Re-evaluation of Shell Egg. Poult Sci 56, 1927 Board. R. G, The microbiology of the Hen's egg. Adv. Appl Microbiol 984; (11) Soeherwin Mangundjaja, Viabilitas Kuman Anaeron dalam telur Ayam.(Viability of the anaerobic bacteria in hen egg) Kongres Perhimpunan Mikrobiologi Indonesia 1989 Ambarukmo Hotel Yogyakarta Presented at The International Association for Dental Research of Asian Pacific Section Singapore