Treatment of ship ballast water with sodium hydroxide (NaOH) is one method currently being developed to minimize the risk to introduce aquatic invasive species. The bactericidal capability of sodium hydroxide was determined for 148 bacterial strains from ballast water collected in 2009 and 2010 from the M/V Indiana Harbor, a bulk-freight carrier plying the Laurentian Great Lakes, USA. Primary culture of bacteria was done using brain heart infusion agar and a developmental medium. Strains were characterized based on PCR amplification and sequencing of a portion of the 16S rRNA gene. Sequence similarities (99+ %) were determined by comparison with the National Center for Biotechnology Information (NCBI) GenBank catalog. Flavobacterium spp. were the most prevalent bacteria characterized in 2009, comprising 51.1% (24/47) of the total, and Pseudomonas spp. (62/101; 61.4%) and Brevundimonas spp. (22/101; 21.8%) were the predominate bacteria recovered in 2010; together, comprising 83.2% (84/101) of the total. Testing was done in tryptic soy broth (TSB) medium adjusted with 5N NaOH. Growth of each strain was evaluated at pH 10.0, pH 11.0 and pH 12.0, and 4h up to 72h. The median cell count at 0h for 148 cultures was 5.20×106cfu/mL with a range 1.02×105–1.60×108cfu/mL. The TSB adjusted to pH 10.0 and incubation for less than 24h was bactericidal to 52 (35.1%) strains. Growth in pH 11.0 TSB for less than 4h was bactericidal to 131 (88.5%) strains and pH 11.0 within 12h was bactericidal to 141 (95.3%). One strain, Bacillus horikoshii, survived the harshest treatment, pH 12.0 for 72h.