188 
Fishery Bulletin 115(2) 
Materials and methods 
Study sites 
Mona Island is a carbonate platform located approxi¬ 
mately 73 km west of Cabo Rojo, Puerto Rico (Fig. 1). 
The Mona and Monito Islands National Reserve ex¬ 
tends from the coast out to 17 km (9 nautical miles); 
a year-round no-take zone encompasses all waters 
around the island within 13 km (7 nautical miles) of 
shore. The island itself is a flat-topped, raised platform 
with continuous vertical cliffs around almost the entire 
perimeter. Along the south-southwestern insular shelf, 
there is a shallow lagoon bordered by coral reefs along 
the seaward end (Frank et al., 1998). The spawning ag¬ 
gregation of black grouper at this site is located along 
the southern shelf edge bordered by a steep reef wall. 
Courtship interactions, ventral rubbing, and courtship 
colorations of black grouper were previously recorded 
at this site in conjunction with courtship-associated 
sound (CAS) production (Scharer et al., 2014). 
Riley’s Hump is a small carbonate reef bank to the 
southwest of the Dry Tortugas within the Tortugas 
South Ecological Reserve, Florida—a fully protected 
marine reserve (Fig. 1). This reef bank is a multispe¬ 
cies FSA site and a year-round no-take zone. The bank 
crest rises to a 30-m depth at its shallowest point, and 
the southern and western edges of the platform are 
composed of a steep reef wall (Weaver et al., 2006). 
This spawning aggregation of black grouper at this site 
is located on a pinnacle off the southwestern corner of 
the crest of the seamount. In previous years, courtship 
interactions of black grouper were documented in com¬ 
bination with recorded CASs along the southern wall 
(Locascio and Burton, 2016). 
Bajo de Sico is a seamount located approximately 27 
km west of Mayagiiez, Puerto Rico (Fig. 1). A seasonal 
marine protected area, it is closed to fishing from 1 
October to 31 March for all reef fish species regulated 
by the Caribbean Fishery Management Council. Con¬ 
sisting of 31.2 km^, this seamount is jointly managed 
by Puerto Rico and U.S. federal jurisdictions. This sea¬ 
mount supports hermatypic corals at depths between 
40 and 90 m and has vertical drop-offs along its west¬ 
ern and northwestern edges. Courtship behavior of 
black grouper was witnessed (senior author, personal 
observ.) in an area characterized by a steep reef wall 
and large rock promontory. 
Passive acoustic recording 
Digital spectrogram recorders (DSG-Ocean^, Logger- 
head Instruments, Sarasota, FL) were deployed at all 
3 sites preceding aggregation of fish (Table 1). One 
digital spectrogram (DSG) recorder was deployed at 
the known FSA location at Mona Island. Multiple DSG 
' Mention of trade names or commercial companies is for iden¬ 
tification purposes only and does not imply endorsement by 
the National Marine Fisheries Service, NOAA. 
Table 1 
Dates of deployment and recovery of autonomous 
acoustic digital spectrogram recorders used for analy¬ 
sis of production of courtship-associated sounds made 
by black grouper (Mycteroperca bonaci) at 2 sites in 
Puerto Rico (Mona Island and Bajo de Sico) and 1 site 
off southern Florida (Riley’s Hump) during 2012-2014. 
Deployment and recovery specifically refer to beginning 
and end dates of time-series data used in analysis. 
Year 
Site 
Deployment 
Recovery 
2012 
Mona Island 
12/20/11 
4/30/12 
Riley’s Hump 
12/18/11 
5/29/12 
2013 
Mona Island 
12/20/12 
4/30/13 
Bajo de Sico 
12/16/12 
4/30/13 
2014 
Mona Island 
12/28/13 
4/30/14 
Bajo de Sico 
12/20/13 
4/30/14 
recorders v/ere deployed around the spawning areas at 
Riley’s Hump and Bajo de Sico. For each of those 2 
sites, data from the DSG recorder with the most CAS 
recordings were used for analysis. High CAS rates, 
high sound levels, and temporal patterns of CASs from 
each selected DSG recorder indicate a strong likelihood 
of close proximity to a spawning location. 
The DSG recorders at Bajo de Sico and Mona Island 
collected 20-s audio clips every 5 min during deploy¬ 
ment (Table 1). At Riley’s Hump, the DSG recorder col¬ 
lected 10-s audio clips every 10 min during deployment 
(Table 1). Differences in these recording schedules re¬ 
quired different corrective factors to extrapolate total 
CASs from sample size to daily totals for comparison. 
Daily total recorded CASs at Riley’s Hump were mul¬ 
tiplied by 60 to calculate the total number of CASs 
per day. Daily total recorded CASs at Bajo de Sico and 
Mona Island were multiplied by 15 to calculate total 
number of CASs per day. After recovery of DSG record¬ 
ers, recorded files were downloaded and converted to 
.wav format for visual analysis with Ishmael software, 
vers. 2.4 (Cooperative Institute for Marine Resources 
Studies Bioacoustics Lab, Oregon State University, 
Newport, OR) (Fig. 2). Any questionable CASs were 
verified audibly by using Windows Media Player and 
noise cancelling headphones. Audible detection of a 
pulse train undulation that was specific to black grou¬ 
per at a frequency band between 75 and 100 Hz served 
as species verification. 
Temporal analysis 
The total number of CASs per day from each DSG 
recorder was calculated for each calendar day during 
deployment. The monthly averages of daily total num¬ 
bers of CASs were compared to confirm seasonality. 
Daily total numbers of CASs were analyzed in rela¬ 
tion to the number of days after a full moon (DAFM) 
to analyze patterns associated with lunar periodicity. 
