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Abstract This study was conducted in Microbiology Department Faculty of Medicine Zagazig University on samples obtained from Haemodialysis Unit at Zagazig University Hospitals. • The aim of this study was detection of Gram negative bacilli by (conventional methods, API and PCR) in addition to detection of the endotoxin by The Limulus Amebocyte Lysate (LAL) assay in water and dialysate of haemodialysis unit in Zagazig University hospitals.• 281 water samples, 50 concentrate samples and 100 dialyste samples were collected under aseptic conditions and with application of all precautions during sampling and transport to get valuable results. All tested samples were cultured on four test media namely ,M- Endo at 37 °C for 24 hours ( selective medium for total coliform ), M- FC medium at 45 °C for 48 hours ( selective for faecal coliform ), R2A medium at 22 °C for 7 days ( for isolation of heterotrophic water contaminants ) and cetermide medium at 37 °C for 24 hours ( selective for Pseudomonas auroginosa . No water samples showed coliform growth while the tested fifty concentrate samples showed 12 % (6/50) and 10 % of 100 dialysate samples showed coliform growth .<br. |