The experiments reported here comply with the current laws of Germany (approved by Regierungspräsidium Darmstadt V-54-19c-20/15-F104/Anz.18) and the USA (approved by the Institutional Animal Care and Use Committee of the University of Oklahoma; AUS-IACUC approved protocols: R06-026 and R09-023).
Test subjects were lab-reared descendants of wild-caught fish. We included Atlantic mollies from the coastal lagoons around the Central Mexican city of Tampico (belonging to the subspecies
P. mexicana limantouri); another population was collected in the Río Oxolotan in Tabasco, South México (
P. mexicana mexicana). Recent phylogenetic analyses argue in favor of full species status of the two subspecies
48 (link). We further included a locally adapted and genetically differentiated (
i.e., independently evolving) ecotype from the
P. mexicana mexicana clade: the hydrogen sulfide-adapted form inhabiting El Azufre, a tributary to the Río Oxolotan
49 (link),
50 (link). As another representative of short-fin mollies
23 (link),
51 (link) we included mangrove mollies (
P. orri) from Roatán Island, Honduras. Two species of long-fin mollies were tested: sailfin mollies (
P. latipinna) stemmed from the Comal River in Central Texas, USA, while Tamési mollies (
P. latipunctata) were collected near Ciudad Mante in Tamaulipas, México. We further included guppies (
P. reticulata) from Venezuela and a feral population from the San Antonio River, Texas, USA
52 , as well as Venezuelan swamp guppies (
P. picta). As representatives of the genus
Limia, we included
L. tridens and sulfur limia (
L. sulphurophilia), both originating from the Dominican Republic.
Gambusia sexradiata from the Río Teapa, and Grijalva mosquitofish (
Heterophallus milleri) from the Río Oxolotán (both Tabasco, México) were included as representatives of mosquito fishes.
Test fish came from large, randomly outbred single-species stocks maintained at the Department of Ecology and Evolution of the University of Frankfurt (
P. m. mexicana,
P. m. limantouri,
P. reticulata from Venezuela,
P. picta,
L. tridens), or at the Department of Zoology at the University of Oklahoma in Norman (
P. m. mexicana from El Azufre,
P. latipinna,
P. latipunctata,
P. orri, feral
P. reticulata,
L. sulphurophila,
G. sexradiata, H. milleri;
Table 1 ). Fish were reared as single-species, mixed-sex stocks in 200-l (Frankfurt) or 1,000-l (Norman) tanks at 25–27°C under an 12:12 hours light: dark cycle (Frankfurt) or under ambient light conditions in a greenhouse (Norman). At the University of Frankfurt, fish were fed twice daily
ad libitum with commercial flake food. Stock tanks in Norman contained naturally growing algae as well as a variety of naturally occurring invertebrates such as chironomid larvae, copepods and amphipods, on which the fish could feed. In addition, fish were supplied with flake food every two days. However, at least 1 week prior to the behavioral experiments, fish were fed
ad libitum at least once daily with flake food.
Test subjects were lab-reared descendants of wild-caught fish. We included Atlantic mollies from the coastal lagoons around the Central Mexican city of Tampico (belonging to the subspecies
P. mexicana limantouri); another population was collected in the Río Oxolotan in Tabasco, South México (
P. mexicana mexicana). Recent phylogenetic analyses argue in favor of full species status of the two subspecies
48 (link). We further included a locally adapted and genetically differentiated (
i.e., independently evolving) ecotype from the
P. mexicana mexicana clade: the hydrogen sulfide-adapted form inhabiting El Azufre, a tributary to the Río Oxolotan
49 (link),
50 (link). As another representative of short-fin mollies
23 (link),
51 (link) we included mangrove mollies (
P. orri) from Roatán Island, Honduras. Two species of long-fin mollies were tested: sailfin mollies (
P. latipinna) stemmed from the Comal River in Central Texas, USA, while Tamési mollies (
P. latipunctata) were collected near Ciudad Mante in Tamaulipas, México. We further included guppies (
P. reticulata) from Venezuela and a feral population from the San Antonio River, Texas, USA
52 , as well as Venezuelan swamp guppies (
P. picta). As representatives of the genus
Limia, we included
L. tridens and sulfur limia (
L. sulphurophilia), both originating from the Dominican Republic.
Gambusia sexradiata from the Río Teapa, and Grijalva mosquitofish (
Heterophallus milleri) from the Río Oxolotán (both Tabasco, México) were included as representatives of mosquito fishes.
Test fish came from large, randomly outbred single-species stocks maintained at the Department of Ecology and Evolution of the University of Frankfurt (
P. m. mexicana,
P. m. limantouri,
P. reticulata from Venezuela,
P. picta,
L. tridens), or at the Department of Zoology at the University of Oklahoma in Norman (
P. m. mexicana from El Azufre,
P. latipinna,
P. latipunctata,
P. orri, feral
P. reticulata,
L. sulphurophila,
G. sexradiata, H. milleri;
ad libitum with commercial flake food. Stock tanks in Norman contained naturally growing algae as well as a variety of naturally occurring invertebrates such as chironomid larvae, copepods and amphipods, on which the fish could feed. In addition, fish were supplied with flake food every two days. However, at least 1 week prior to the behavioral experiments, fish were fed
ad libitum at least once daily with flake food.
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