Neonates in intensive care units often require supporting medical devices and antibiotic treatment. The intensive care treatment combined with their immature immune system, the increased permeability of mucosa, and the undeveloped microflora of the gut may render the neonates highly vulnerable to colonisation and subsequent infections when exposed to opportunistic pathogens. These infections may not only be local gastrointestinal infections, but also systematic following translocation from the gastrointestinal system. This could be particularly alarming considering that common antibiotics may not be effective if the causative strain is multi-drug resistant.This chapter reviews our information on the microbial colonization of neonatal feeding tubes. The range of organisms which have been recovered are wide, and while primarily bacterial, fungi such as Candida have also been found. The bacteria are principally Staphylococcus spp. and Enterobacteriaceae. The Enterobacteriaceae isolates are predominantly Enterobacter cancerogenus, Serratia marcescens, Enterobacter hormaechei, Escherichia coli and Klebsiella pneumoniae. Many of these isolates encode for antibiotic resistance; E. hormaechei (ceftazidine and cefotaxime) and S. marcescens strains (amoxicillin and co-amoxiclav).