A cross-sectional study was conducted in backyard poultry flocks among nine counties of Maryland from May 2011 to August 2011. The objective of this study was to obtain baseline data from a survey on biosecurity practices and investigate risk factors associated with positive findings of avian influenza (AI), Newcastle disease (ND), infectious laryngotracheitis (ILT), Mycoplasma gallisepticum (MG), and Salmonella Enteritidis (SE). Serum, tracheal, and cloacal swabs were randomly collected from 262 birds among 39 registered premises. Analysis revealed flock prevalence and seroprevalence respectively for the following: AI (0%, 23%), ND (0%, 23%), ILT (26%, 49%), MG (3%, 13%), SE (0%, ND). Vaccine status could not be confirmed for ND, ILT, or MG. Premises positives were identified by partial nucleotide sequencing. No statistically significant associations were identified, however, AI seroprevalence was positively associated with exposure to waterfowl (Relative Risk [RR] = 3.14, 95% confidence interval [CI] 1.1-8.9) and absence of pest control (RR=2.5; 95% CI, 0.6-10.4).