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BACKGROUND: Salmonella and Shigella remain the majorcontributors to acute enteric infections and diarrhoea. Hence, theobjective of this study was to isolate and determine theantimicrobial susceptibility pattern of Shigella and Salmonellaspecies from children with acute diarrhoea in Mekelle Hospital andSemen Health Center.METHODS: A cross sectional study was conducted among 260children with acute diarrhoea from November 2011 to March 2012in Mekelle, Ethiopia. Stool specimen was collected from all studyparticipants who presented with acute diarrhoea. Microscopy,culture and confirmatory identification were done by the pattern ofbiochemical reactions using a standard bacterial identificationsystem (API 20E, BioMerieux, Marcy-l’Etoile, France) andpolyvalent (Poly O and H) antiseras for Salmonella species and Vifor S.typhi. Isolated colonies were assessed for antimicrobialsusceptibility profile using disk diffusion method. Data was enteredand analyzed using SPSS version 16.0 software.RESULTS: Out of the 260 study participants, 145(55.8%) weremales while 115(44.2%) were females. The majority of the patients(44.2%) were of children under five years old. A total of 120enteropathogens were isolated. The frequency of isolation was19(7.3%), 18(6.9%) and 83(31.9%) for Salmonella species, Shigellaspecies and intestinal parasites respectively. Most of the Shigellaisolates were resistant to ampicillin (88.9%), Tetracycline (77.8),cotrimoxazole (55.6%) and chloramphenicol (55.6%). Among theSalmonella isolates, the highest resistance was observed toampicillin (89.5%), Tetracycline (89.5%), chloramphenicol (78.9%)and cotrimoxazole (57.9%). Multi-drug resistance was noted in19(100%) and 16(88.9%) of Salmonella and Shigella speciesrespectively.CONCLUSIONS: Shigella and Salmonella are still challengingpathogens in children < 5 years of age. High antibiotic resistancewas observed among both isolates to ampicillin, tetracycline,chloramphenicol and cotrimoxazole.