Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
2Laboratory of Microbiology, Facult y of Veterinary Medicine, Syiah Kuala Universit y, Banda
Aceh 23111, I ndonesia
3Laboratory of Veterinary Public Healt h, Facult y of Veterinary Medicine, Syiah Kuala Universit y,
Darussalam , Banda Aceh 23111, I ndonesia;
4*Laboratory of Pathology, Facult y of Vet erinary Medicine, Syiah Kuala Universit y,
Darussalam , Banda Aceh 23111, I ndonesia;
5Laboratory of Parasit ology, Facult y of Veterinary Medicine, Syiah Kuala Universit y, Banda
Aceh 23111, I ndonesia;
Ke y w or ds: Salm onella sp., chloram phenicol, gent am icin, t et racy cline, pineapple squeeze
I n t r od u ct ion 36° C for 48 hours. Cult ures are exam ined each day for growt h and any colonies are Gram st ained and subcult ured ( i.e., t ransferred) t o appropriate m edia as explained by Darm awi et al., 2013) .
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
Gr a m St a in
I n t his study we obt ain a clean glass slide to prepare a sm ear of each bacteria t o be st ained by t aking a loopful of t he bact eria (wit h a st erile loop) and spreading it over a sm all area in t he cent er of the slide. The slide allowed t he sm ear t o air dry and t hen heat fix by passing t he slide quickly t hrough a flam e. The slide placed on paper t owels and add a drop or t wo of cryst al violet t o t he sm ear t o let set 1 m inute. The slide washed gent ly t he st ain off wit h t ap water carefully in order t o being not to wash off bact eria. Then, Gram ’s iodine apply t o let set 1 m inute. To rem ove any excess st ain or st ain that has not adhered t o t he cell, t he slide washed gent ly t he iodine off wit h t ap wat er and t hen add t he decolorizing agent ( 95% Et OH) drop by drop unt il it runs clear. The decolorizing reagent washed off wit h t ap wat er, and count erst ain wit h safranin by adding 1- 2 drops and let it set for 45 seconds. Finally, t he slide rinsed with tap water, looked at under t he m icroscope, and det erm ined if bact erium is Gram - posit ive or Gram - negat ive. Gram - negat ive cells will pick up t he counterst ain and appear red or pink as described by previously st udy ( Health Prot ect ion Agency, 2007; Darm awi et al., 2011; Darm awi et al., 2013) wit h certain m odificat ion.
Bioch em ica l Assay ed
All isolat es were ident ified according t o t he m et hods advocated by Edwards and Ewing ( 1962) . The specific m et hods involved were colonial characteristics on m edia including size, inabilit y t o swarm , abilit y or inabilit y t o ferm ent lactose. Specific t est s such as carbohydrate nam ely: Mannit ol, Sucrose, Lactose, Glucose, Triple Sugar I ron Agar, indole form at ion, Methyl Red ( Oxoid) , Voges Proskauer ( Oxoid) , and Sulfic I ndole Mot ilit y, Sim m on’s Cit rate ( Oxoid) t est s were done as described by Darm awi
et al., 2011; Darm awi et al., 2013) wit h cert ain m odificat ion.
Pr ep e r a t ion of Pin e ap ple Sq u e ez e
Pineapple (A. com osus) were procured from local m arket ( Banda Aceh, I ndonesia) . The crude equeous squeeze of t he pineapple fruit s were prepared by pineapple squeezer.
Kir by - Bau e r An t ibiot ic Te sting
Cult ures were com pared wit h Mc Farland 3 st andardizat ion, and separated on surface of Muller Hint on Agar ( MHA). Organism s are also test ed for ant ibiot ic suscept ibilit y by t he Kirby Bauer m et hod. Ant ibiot ic sensit ivit y was tested by m ean of choram phenicol and gent am ycin. The ant ibiot ic discs ( Oxoid) and pineapple ext ract were put on t he surface of MHA m edia which had inoculated wit h
Salm onella sp. as described by Darm awi et al., 2011; Darm awi et al., 2013) wit h certain m odificat ion. The blank ant ibiot ic disks ( Oxoid) were put on the cent er, and the flooded pineapple squizee and ant ibiot ic discs were put in t he surface of MHA m edia which had inoculated wit h bacteria. After overnight incubat ion, t he zone of inhibit ion is appear on t he surrounding of disck.
Resu lts a nd D iscussion
The sam ples were posit ive for spesies of the Salm onella sp. bacteria isolat ed from feces of broiler chickens as shown in Table 1.
Table 1. Salm onella sp. bact eria isolated from feces of broiler chickens
Biochem ical Assayed
Test Mannitol Sucrose Lactose Glucose SI M TSI A Sim m on cyt rate
MR VP I ndol
React ion + - - + + + + + - +
Cult ure separated on surface of Mac Conkey or Brilliant Green Agar develop bact eria colony. Under m icroscope, Gram - negat ive cells appeared red or pink colour as shown in Figure 1A. The zone form ed in t his m anner is inhibit Salm onella sp. growth shown in Figure 1B.
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
Table 2. Diam eter inhibit ion zone of ant ibiot ics and pineapple squeeze
Ant ibiot ic Zone (m m ) course of t he salm onellosis. Depending on t he severity and t he t ype of bacteria, t he ant ibiot ic m ay be given. I f t he bact eria are suscept ible t o a part icular ant ibiot ic, an area of clearing surrounds t he m edicine include penicillin, cephalosporins, am inoglycosides, m acrolides, quinolones and tet racyclines. Do not t ake ant ibiot ics for longer t han needed. Long term use of ant ibiot ics leads to resistance and chances of developing oport unist ic infect ions.
Con clusions
I nhibit ion growt h of Salm onella sp. caused by chloram phenicol and gent am icin was significant ly higher in com parison wit h pineapple (Ananas com osus) squeeze, m eanwhile t he Salm onella sp. bact eria was resist ance against t et racycline.
Ack n ow led g em e n ts
We wish t o t hank t he chairm an and all t heacing staff of Microbiology Laborarory, Facult y of Veterinary Medicine of Syiah Kuala Universit y t hat frendly pleased us t o conduct t his research.
Proceeding s of Th e 5th Annu al I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2 01 5
I n conj unct ion w it h The 8th I n t ern at ion al Con feren ce of Ch em ical Eng ineering o n Science and Applicat ions ( Ch ESA) 2 0 15 Sept em ber 9 - 1 1, 2 01 5, Banda Aceh, I ndon esia
Re fe r e n ce s
Andrew s, J. M. 200 1. Det erm inat ion of m in im um inhibit ory concent rat ions. Journal of Ant im icrobial Chem ot herapy 48 ( Suppl. 1) : 5- 16, PMI D 114203 33.
Darm aw i, Darniat i, Zak iah Hery aw at i Manaf, Sy afruddin and Arm an Say ut i. 2011. I solat ion of Gram - Negat iv e Bact eria from Met acarpal I n j ury of Pant hera t igris su m at rae Trapped in Subulussalam , I ndonesia. Proceedings Annual I nt ernat ional Conference ( AI C) , I nt ern at ional Sem inar, Sy iah Kuala Univ ersit y , Banda Aceh. 29- 30t h Nov em ber 2011. Hal. 13 1- 135, I SSN: 2089- 2 08X.
Darm aw i, E. Rahm i, M. Dew i, J. Hut abarat , M. Abrar an d Fak hrurrazi. 20 13. I solat ion of ent erobact er iaceae bact eria species from feces of Sum at ran Orangut ans (Pongo abelii) . Proceedings t he 3rd Annual I nt ernat ional Conference Sy iah Kuala Univ ersit y ( AI C Unsy iah) in Conj unct ion w it h t he 2nd I nt ernat ional Conference on Mult idisciplinary Research ( I CMR) , Banda Aceh. 2- 4t h Ok t ober 2013. Hal. 349- 354,I SSN: 2089- 208X.
Goldst ein EJ, Cit ron DM, Finegold SM. 1984. Role of anaerobic bact eria in bit e- w ound infect ions. Rev I nfect Dis. Suppl 1: S177- 83. ht t p: / / w w w .ncbi.nlm .nih.gov / pubm ed/ 63 72026 ( 27- 0 3- 201 1)
Turnidge JD, Ferrar o MJ, Jorgensen JH. 2003 . Suscept ibilit y Test Met hods: General Considerat ions. I n PR Murray , EJ Baron, JH Jorgensen, MA Pfaller, RH Yolk en. Manual of Clinical Microbiology . 8t h Ed. Washingt on. Am erican Societ y of Clinical Microbiology . p 1103 I SBN 1- 55 581- 255- 4
