PRISUSTVO KOAGULAZA POZITIVNIH STAFILOKOKA KOD PASA
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1 Igor Stojanov, Sara S avic-jevdenic, Etjen Mišic Naucni institut za veterinarstvo, Novi Sad Veterinarska ambulanta PRISUSTVO KOAGULAZA POZITIVNIH STAFILOKOKA KOD PASA PRESENCE OF COAGULASE POSITIVE STAPHYLOCOCCI IN DOGS ABSTRACT: Coagulase positive staphyloccoci present microorganisms which, as it can be seen in the name they carry (enzyme coagulase), and belong to a pathogen group of microflora organisms. Among them the most important are Staphylococcus aureus and Staphylococcus intermedius. The objective of this paper is bacteriological monitoring on the presence of some coagulase positive staphyloccoci in dogs. The aim was to determine how much is coagulase positive staphyloccoci present in dogs and to point a possibility that pets may transfer microorganisms on people. The results point on the presence of these bacteria in humans and in pets, where dogs may transmit bacterial infection on humans, or they can be transsmitted from humans to dogs. KEY WORDS: man, dogs, coagulase positive staphyloccoci UVOD Rod Staphylococcus je široko rasprostranjen u prirodi i može se naci i kod ljudi i kod mnogih drugih vrsta sisara i ptica. Iz ovog roda, po znacaju za zdravstveno stanje ljudi i životnja, svakako treba izdvojiti koagulaza pozitivne stafilokoke od kojih cemo spomenuti Staphylococcus aureus, Staphylococcus intermedius i za veterinu znacajan Staphylococcus hyicus. Mesta na kojim se mogu naci kod ljudi kao što su sluzokoža gornjih delova respiratornog trakta, površini kože, i delovima urogenitalnog trakta (Karakaševic i sar. 1989), gotovo u potpunosti odgovaraju mestima na kojima se mogu naci i kod pasa kucnih ljubimaca, koji su, cini se, postal! sastavni deo covekove prirodne sredine u urbanim uslovima življenja. Koagulaza pozitivne stafilokoke predstavaljaju mikrofloru koja poseduje enzim koagulazu, a prisustvo ovog enzima je najsigurniji znak patogenosti mikroorganizma. Korišcenje samo koagulaza testa nije dovoljno za razlikovanje dve najznacajnije 263
2 vrste St. aureus i St. intermedius na šta ukazuje rad (Vandenesch F. i sar. 1995). Predmet našeg rada je bakteriološki monitoring prisustva nekih od koagulaza pozitivnih stafilokoka kod pasa, sa osvrtom na mesta na kojima se mogu naci i vrstama stafilokoka sa ovim karakteristikama. Cilj nam je da utvrdimo kolika je rasprostranjenost koagulaza pozitivnih stafilokoka kod pasa, o kojim se vrstama radi, koja su najcešca mesta na kojima se nalaze i da se ukaže na mogucnost uloge kucnih Ijubimaca" kao vektora prenošenja ovih mikroorganizama na ljude. MATERIJAL I METODE U radu su prezentovani podaci obradenih materijala koji su prispeli u našu laboratoriju tokom jedne godine. Ukupno je obradeno 253 materijala. Materijal koji je obradivan bio je poreklom od pasa koji su imali razlicite klinicke nalaze ili kod kojih su postojali zdravstveni prsblemi u dužem vremenskom periodu. Ipitivani materijal je bio raznolik: brisevi sa kože psa (sa razlicitih delova tela), brisevi oka, grla, nosa, uha, vaginalni brisevi, brisevi prepucijuma, mleko, urin i sperma. U toku rada koristili smo standardnu laboratorijsku metodologiju. Izolacija mikroorganizama je obavljena direktnim zasejavanjem materijala na krvni agar (10% defibrisane ovcije krvi) i selektivno diferencijalnu podlogu XLD (ksiloza lizin deoksiholat agar) (Q i u n n J. P. i sar. 1998), dok je identifikovanje utvrdeno odredivanjem fizioloških karakteristika koje su osobene za koagulaza pozitivne stafilokoke. REZULTATI I DISKUSIJA Kako je vec spomenuto, prisustvo koagulaza enzima govori o patogenosti koagulaza pozitivnih stafilokoka. Humani sojevi stafilokoka dovode do koagulacije plazme kunica i ljudi dok, animalni pored navedenih mogu koagulisati i plazmu goveda, ovaca i svinja. (Q i u n n J. P. i sar. 1998). Pored spomenutog enzima patogenost stafilokoka se ogleda i u prisustvu drugih toksina i enzima. Pre svega enterotoksina (A E) koji imaju veliki znacaj kod trovanja hranom ljudi i nadražaja centra za povracanje, eksfoliaciju produkuju stafilokokni sindrom kožnog perutanja kod dece i narocito kod pasa (SSSS), humani toksicni sindrom (TSS) prouzrokovan toksinom TSS-1, epidermolitican toksin koji je u vezi eksuditavnog epidermitisa prasadi, a moguca je produkcija i pirogenog toksina. Produkciju enzima ukljucuje i prisustvo alfa i beta toksina, stafilokinazu, hijaluronidazu, lipazu, kolagenazu proteazu, nukleazu i ureazu od kojih svi mogu imati ulogu u patogenosti ovih mikroorganizama ( Q i u n n J. P. i sar. 1998). Izolate koji su davali potpunu beta hemolizu, na krvnom agaru, neposredno oko samih kolnija i nepotpunu hemolizu na delu gde se završavala potpuna i nisu imali rast na XLD-u bojili smo po Gramu. Gram pozitivni sojevi koji su imali oblik grozda u preparatu testirani su oksidaza i katalaza testom. Katalaza pozitivni i oksidaza negativni sojevi koji su u OF medijumu pokazali fermentativne osobine tesirani su koagulaza testom u epruveti (korišcena je plazma kunica) i sledecim biohemijskim karakteristikama (prisustvo ureaze, fermentacije manita i maltoze, eskulin hidrolize, ostljivost na novobiocin (5 pig disk) i osetljivost na polimiksin (300 unit disk) (Q i u n n J. P. i sar. 1998). 264
3 U Tabeli 1. dati su podaci uradenih testova koji su nam, u odnosu na literaturne podatke, dali odgovor o kojoj se vrsti koagulaza pozitivnih stafilokoka radi. Tabela 1. Fiziološke karakteristike koagulaza pozitivnih stafilokoka Staphylococcus aureus Staphylococcus intermedius Staphylococcus hyicus Koagulaza test + + D Hemoliza + + Produkcija pigmenta +* Ureaza D + D Fermentacija manitola + D Fermentacija maltoza + (+/-) Hidroliza eskulna Osetljivost na novobiocin (5 mg disk) S S S Osetljivost na polimiksin B (300 unit disk) R S R * sojevi izolovani kod pasa su obicno bez pigmenta D 11 89% pozitivna reakcija (+/-) 90% i vise slabo pozitivni S osetljiv R rezistentan (rezistencija za novobiocin je zona od 16 mm ili manja, rezistencija za polimik - sin B je 10 mm ili manja) U toku ispitivanja prispelih materijala poreklom od pasa utvrdeno je prisustvo koagulaza pozitivnih stafilokoka u svim vrstama materijala koji su stigli tokom jedne godine. Rezultati ovih analiza se nalaze u Tabeli 2. Tabela 2. Rezultati ispitivanja primljenog materijala na prisustvo koagulaza pozitivnih satfilokoka Vrsta materijala Obradeni materijali Pozitivan nalaz na koagulaza-pozitivne stafilokoke (% obradenog i pozitvnog) Brisevi kože ,35% Bris uha ,66% Bris grla ,23% Bris nosa ,00% Bris oka ,33% Vaginalni bris ,26% Bris prepucijuma ,00% Mleko ,14% Urin ,00% Sperma ,22% U Tabeli 2. se nalaze podaci koji se odnose na prisustvo Satphylococcus intermedius, odnosno izolate koji su svojim fiziološkim karakteristikama odgovarali drugoj koloni Tabele 1. Svi izolati, sem jednog, koji je izolovan sa površine kože pripadali su St. intermedius. Jedini izolat koji nije pripadao ovoj vrsti je St. aureus i odgovarao je karakteristikama datim u koloni jedan Tabele
4 Znacaj ovakvog nalaza koagulaza pozitivnih stafilokoka, a pre svega St. intermedius ogleda se u rezultatima koji govore o nalazu St. intermedius kod humanih izolata. Tako se u radovima (Mahoudeau i sar. 1997) govori o nalazu St. intermedius izolovanog kao deo nazalne flore i bakteriološkog izolata pleuralne tecnosti. Nje-govo prisustvo je utvrdeno i kod infekcije nastale prilikom aplikovanja lekova kate-terom kod pacijenta koji je bio na lecenju u bolnici (Kandenesch F. i sar. 1995). Isto tako o nalazu St. intermedius, mada sa nedovoljno jasno utvrdenim znaca-jem govori rad (B e s M. i sar. 2002) gde je utvrdeno njegovo prisustvo kao redeg izolata kod ljudi iz rana nastalih kao posledica ujeda pasa. O znacaju nalaza St. intermedius kao dela gingivalne bakterijske flore ukazuje rad (S a r h i r D. A. i sar. 1976) u kome se istice da je poznavanje mikroflore usta važan preduslov na osnovu kojeg se može doneti procena posledica nakon ujeda psa. Kljucni" primer možda je znacaj nalaza koagulaza pozitivnih stafilokoka kod ljudi i životinja predstavlja rad (T a 1 a n D. A. i sar. 1989) koji je analizirao humanu nazalnu mikrofloru i to pre svega veterinara i radnika u veterinarskoj praksi. Utvrdeno je da St. intermedius predstavlja oportunu patogenu zoonoznu infekciju. Ono što se namece kao zaklju-cak iz dobijenih nalaza ovoga rada je cinjenica da bi ovakvi rezultati, možda, bili slic-ni i kada bi se analizirala bakterijska flora vlasnika pasa kucnih Ijubimaca. Naš bakteriološki nalaz je u skladu sa nalazom (K r o g H. V. i sar. 1976) koji odgovara i po nalazu koagulaza pozitivnih stafilokoka i po nalazu ostalih izolovanih mikroorganizama. U Tabeli 3. se nalaze podaci koji ukazuju koliki je broj pozitivnih nalaza na koagulaza pozitivne stafilokoke bio pojedinacan, a koliko se radilo o mešanoj bakterijskoj flori. Tabela 3. Rezultati ispitivanja primljenog materijala na prisustvo koagulaza pozitivnih stafilokoka u odnosu na mešovite izolate I v t... Pozitivan nalaz na mesanjh izolata Vrsta matenjala koa^a-poativne {% pozi^vnih/mesanih izolata) Brisevi kože ,4% Bris uha ,0% Bris grla ,3% Bris nosa ,3% Bris oka ,0% Vaginalni bris ,2% Bris prepucijuma ,0% Mleko ,0% Urin 1 Sperma ,11% U toku ispitivanja izolovan je veci broj razlicitih mikoorganizama. Najcešci mešani izolati cinili su pored koagulaza pozitivnih stafilokoka sledeci mikroorgnaizmi: Streptococcus beta hem. (G grupa po Lancefield -u), Staphylococcus epidermidis, Streptococ - cus alfa hem., E. coli, Proteus mirabilis, Pseudomonas aeuruginosa, Pasteurella multocida, Bacillus spp. Medun'm i u veterinarskoj medicini kao i u humanoj, postoje dileme o znacaju nalaza Staphylococcus intermedius na zdravstveno stanje inh'cirane jedinke. U istraživanjima (Greene. R. T. i sar. 1993) postavlja se pitanje da li njihovo prisustvo samo po sebi izaziva zdravsrvene smetnje ili ono predstavlja sekundarne infekcije koje 266
5 posle primarno izazvanih poremecaja zdravstvenog stanja dodatno pogorsavaju situaciju. Za delimicno razjašnjenje ove dileme može nam pomoci rad (M c e w a n A. N. i sar. 2000, A i e 11 o S. i sar. 1998) koji istice da je adherencija St. intermedius za keratocite statisticki znacajno veca kod životinja sa atopicnim dermatitom (primarna seborea i bakterijska pioderma). Ovakvi rezultati ukazuju da su alergijske reakcije znacajan preduslov za naseljavanje ovih mikoorganizama, i s obzirom na uoceno povecanje alergija kod pasa gajenih u prirodnim uslovima grada" i cinjenicu da alergije predstavljaju bolest modernog doba covecanstva, one mogu dovesti do pojave bolesti inficiranih jedinki. Kod pasa naseljavanje Staphylococcus intermedius može dovesti do pojave razliciti bolesti od kojih demo samo neke važnije spomenuti: stafilokokna enterotoksemija stenadi, metritis, otitis externa, eczema (razucitih vrsta), urethritis, pharyngitis et tonsilitis (Teodosic S. 1994). ZAKLJUCAK 1. U odnosu na vrstu i broj primljenih materijala procentualno najveci broj koagulaza pozitivnih stafilokoka bio je kod analiziranih uzoraka oka (83,33%), zatim briseva kože (77,35%) i briseva grla (69,23%), ali kada se sagleda najveci broj pojedinacnih materijala koji su stigli u laboratoriju, a to su vaginalni brisevi i brisevi kože, može se zakljuciti da su poremecaji na ovim delovima organizma bili najcešci problem pasa, pa tako i mikrobioloski nalaz dobija na znacaju. 2. Odnos izolovanih koagulaza pozitivnih stafilokoka u cistoj kulturi i u mešanim izolatima (najveci procenat mešanih izolata je kod vaginalnih briseva 51,2% i jedva prelaza polovinu ukupno obradenih materijala) ukazuje da se u velikom procentu ove bakterije nalaze kao jedini izolat, što može dovesti do zakljucka da se njihovo prisustvo može smatrati uzrokom nastalog zdravstvenog poremecaja. 3. Ukupna izolovana bakterijska flora odgovara nalazima do kojih su došli i drugi autori koji su pratili slicnu problematiku. 4. Nalaz St. intermedius kod veterinara i radnika u veterinarskoj praksi ukazuje da se slicni nalaz može ocekivati i kod vlasnika pasa i njihovih kucnih ljubimaca, i da ljudi kao i psi mogu predstavljati potencijalne vektore prenošenja koagulaza pozi tivnih stafilokoka. 5. Utvrdeni problemi vezani za adherenciju Staphylococcus intermedius kada je u pitanju imunološka preosetljivost nastala kao posledica senzibilizacije hranom ili drugim alergenima ukazuje na znacajne probleme koji mogu nastati kombinacijom razlicitih agenasa. LITERATURA Aiello Susan,Editor; The Merck veterinary manual. Eight Edition, Merck & Co., INC. Whitehouse Station, N.J., USA, Greene, R T; Lammler, C: Staphylococcus intermedius: current knowledge on a pathogen of veterinary importance, Zentralblatt Fur Veterinarmedizin. Reihe B. Journal of Veterinary Medicine. Series B, Volume 40, Issue 3, May 1993, Pages Karakaševic B. i saradnici; Mikrobiologija i parazitologija, Medicinska knjiga, Beograd Zagreb,
6 Krogh, H V; Kristensen, S: A study of skin diseases in dogs and cats. II. Microflora of the normal skin of dogs and cats, Nordisk Veterinaermedicin, Volume 28, Issue 9, September 1976, Pages Mahoudeau, I; Delabranche, X; Prevost, G; Monteil, H; Piemont, Y: Frequency of isolation of Staphylococcus intermedius from humans, Journal of Clinical Microbiology, Volume 35, Issue 8, August 1997, Pages MCEWAN N. A.: Adherence by Staphylococcus intermedius to canine keratinocytes in atopic dermatitis, Research in Veterinary Science, Volume 68, Issue 3, June 2000, Pages Michele Bes and Yvonne Brun: Staphylococcus: actualites taxonomiques et identification; Revue Francaise des Laboratoires, Volume 2002, Issue 343, May 2002, Pages Prirucnik za laboratorijsku dijagnostiku: Savez veterinara i veterinarskih tehnicara Jugoslavije, OZID, Beograd, Quinn J.P., Carter E.M., Markey B., Carter R.G.: Clinical Veterinary Microbiology; Mosby, London, Philadelphia, St. Luis, Sydney, Tokyo, Saphir, D A; Carter, G R: Gingival flora of the dog with special reference to bacteria associated with bites, Journal of Clinical Microbiology, Volume 3, Issue 3, March 1976, Pages Talan, D A; Staatz, D; Staatz, A; Overturf, G D: Frequency of Staphylococcus intermedius as human nasopharyngeal flora, Journal of Clinical Microbiology, Volume 27, Issue 10, October 1989, Page Teodosic, S.: Bolesti pasa sa hirurškim intervencijama. Izdavac Autor, Niš, Vandenesch, F; Celard, M; Arpin, D; Bes, M; Greenland, T; Etienne, J: Catheter-related bacteremia associated with coagulase-positive Staphylococcus intermedius Journal of Clinical Microbiology, Volume 33, Issue 9, September 1995, Pages
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