In premise plumbing, microbial water quality may deteriorate under certain conditions, such as stagnation. Stagnation results in a loss of disinfectant residual, which may lead to the regrowth of microorganisms, including opportunistic pathogens. In the present study, microbial regrowth was investigated at eight faucets in a building over four seasons in one year. Water samples were obtained before and after 24 h of stagnation. In the first 100 mL after stagnation, total cell counts measured by flow cytometry increased 14- to 220-fold with a simultaneous decrease in free chlorine from 0.17–0.36 mg L
–1
to <0.02 mg L
–1
. After stagnation, total cell counts were not significantly different among seasons; however, the composition of the microbial community varied seasonally. The relative abundance of
Pseudomonas
spp. was dominant in winter, whereas
Sphingomonas
spp. were dominant in most faucets after stagnation in other seasons. Opportunistic pathogens, such as
Legionella pneumophila
,
Mycobacterium avium
,
Pseudomonas aeruginosa
, and
Acanthamoeba
spp., were below the quantification limit for real-time quantitative PCR in all samples. However, sequences related to other opportunistic pathogens, including
L. feeleii
,
L. maceachernii
,
L. micdadei
,
M. paragordonae
,
M. gordonae
, and
M. haemophilum
, were detected. These results indicate that health risks may increase after stagnation due to the regrowth of opportunistic pathogens.