Neoechinorhynchus (Neoechinorhynchus) schmidti Barger, Thatcher & Nickol, 2004

(Fig. 2 i)

Mexico: TABASCO: 24 km to S of Villahermosa; Trachemys venusta (Barger 2004; Barger et al. 2004). Pantanos de Centla (18°28’18.9’’N 92°39’14.9’’W). Trachemys scripta (Martínez-Aquino et al. 2009; García-Varela et al. 2011).

Specimens deposited. CNHE (6764), HWML (17667), USNPC (92883).

Species delimitation through phylogenetic analysis and genetic divergence. To demonstrate the species boundaries among the species reported in this checklist, we sequenced a total of 269 individuals of the species: N. (N.) brentnickoli (81 individuals from 16 localities); N. (N.) roseum (two individuals from two localities); N. (N.) mamesi (22 individuals from seven localities); N. (N.) golvani (80 individuals from 11 localities); N. (N.) mexicoensis (47 individuals from eight localities); N. (N.) schmidti (five individuals from one locality); N. (N.) emyditoides (26 individuals from three localities); N. (N.) panucensis (four individuals from two localities); and N. (N.) chimalapasensis (two individuals from one locality). Additionally sequence of N. saginata plus Floridosentis mugilis and Floridosentis pacifica were aligned together, composing a data set of 806 bp with 274 terminals. Nucleotide frequencies for 28S rDNA data set were 0.246 (A), 0.178 (C), 0.276 (G), and 0.298 (T). Phylogenetic analyses of the available sequences for the genus Neoechinorhynchus through the Bayesian consensus tree yielded the same branch pattern as the ML tree (Fig. 3). Both trees yielded that the ten species of Neoechinorhynchus are monophyletic with strong nodal support ranging from 72 to 100 % of bootstrap, and 1.0 of posterior probability (Fig. 3). Both trees yielded three major clades. The first clade contained four species N. (N.) roseum, N. saginata, N. (N.) schmidti and N. (N.) emyditoides . The second clade was composed of two species N. (N.) golvani and N. (N.) panucensis and had 100 % bootstrap support and 1.0 posterior probability. The third clade was composed of four species N. (N.) chimalapasensis, N. (N.) brentnickoli, N. (N.) mamesi and N. (N.) mexicoensis and received reliable bootstrap support and high values of posterior probability (Fig. 3).

67- 76 N. (N.) brentnickoli 10 D. latifrons Laguna de Tres Palos, Guerrero, México FJ388991, 8178 FJ968156 - FJ968159, KR086219 - KR086223 *

77- 81 N. (N.) brentnickoli 5 D. latifrons Río Tamarindo, Chautengo, Guerrero, KR086224 - KR086228 * 8179 México

82 N. (N.) roseum 1 Achirus mazatlanus Laguna el Caimanero, Sinaloa, México FJ388999 6762, 633, 634 83 N. (N.) roseum 1 Citharichthys gilberti Estero La Tovara, Nayarit, México FJ389000 6763 84- 86 N. (N.) mamesi 3 D. latifrons Puente Manialtepec, Oaxaca KR086229 - KR086231 *

......continued on the next page TABLE 1. (Continued)

Samples Species N Host Locality GenBank Specimensdeposited

(DNA) CNHE

87- 91 N. (N.) mamesi 5 D. latifrons Estero Joaquin Amaro, Chiapas, México JN830770 - JN830774 8183

92- 95 N. (N.) mamesi 4 D. latifrons Laguna la Conquista, Chiapas, México JN830766 - JN830769 8184

96- 98 N. (N.) mamesi 3 D. latifrons Laguna Rion Pijijiapan, Chiapas, JN830763 - JN830765 8180, 8181, 8182

México

99- 100 N. (N.) mamesi 2 D. latifrons Quebrada Ganados, Costa Rica KR086232 *, KR086233 * 8590

101- 102 N. (N.) mamesi 2 D. maculatus Playa Grande, Costa Rica KR086234 *, 8191 KR086235 *

103- 105 N. (N.) mamesi 3 D. latifrons Las lisas, Guatemala KR086236 - KR086238 *

106- 107 N. (N.) golvani 1 Parachromis Lago Jalapa, Costa Rica KR086239 *

managuensis

108- 109 N. (N.) golvani 2 Parachromis loisellei Lago Jalapa, Costa Rica KR086240 *,

KR086241 *

110- 111 N. (N.) golvani 2 Amphilophus Lago Jalapa, Costa Rica KR086242 *, KR086243 * 8592 longimanus

112 N. (N.) golvani 1 Heterotilapia Lago Jalapa, Costa Rica KR086244 *

multiespinosa

113- 114 N. (N.) golvani 2 Archocentrus Lago Jalapa, Costa Rica KR086245 *,

centrarchus KR086246 *

115- 116 N. (N.) golvani 2 Amatitlania Quebrada Puercos, Costa Rica FJ388998, 6757 nigrofasciata KR086247 *

117- 127 N. (N.) golvani 11 Vieja pearsei Presa Chicoasen, Chiapas, México FJ388995, FJ968136 - FJ968138, 6755 KR086248 - KR086254 *

128- 132 N. (N.) golvani 5 V. pearsei Presa Nezahualcoyolt, Chiapas, México FJ388996, FJ968141, FJ968142, 6756 KR086255 *,

KR086256 *

133- 143 N. (N.) golvani 11 Parachromis Lago Canitzan, Tenosique, Tabasco, FJ388994, FJ968139, FJ968140, 6767 friedrichstalii México KR086257 - KR086264 *

144- 153 N. (N.) golvani 10 Cichlasoma Río Carrizal, Tabasco, México FJ388993, FJ968134, FJ968135, 6754 urophthalmum KR086265 - KR086271 *

154- 156 N. (N.) golvani 3 C. urophthalmum Lago las Ilusiones, Tabasco, México FJ388992 - FJ968144

157- 167 N. (N.) golvani 11 Paraneetroplus Lago de Catemaco, Veracruz, México FJ388986, FJ968145, FJ968146, 601, 603, 604, 606, 631,

fenestratus KR086272 - KR086279 * 632, 6783

168- 172 N. (N.) golvani 5 C. urophthalmum Presa Temascal, Oaxaca, México KR086280 - KR086284 * 8593

......continued on the next page TABLE 1. (Continued) The sequence divergence estimated among the nine species herein considered, plus an additional sequence available for a species that occurs in freshwater fishes in North America ( N. saginata) ranged from 3.2% to 48.7% (Table 2). Instead, the intraspecific genetic divergence varied markedly among all the isolates sequenced for each acanthocephalan species and it was never higher than 3% for all intraspecific comparisons. For instance, the divergence among 81 specimens of N. (N.) brentnickoli obtained from a wide geographic range, in 16 localities (localities 1-16 in Fig. 1) across the Pacific coast of northwestern Mexico, varied from 0 to 0.7% (Table 2). In other example, N. (N.) roseum corresponding to two individuals of two localities 1) Laguna el Caimanero, Sinaloa and 2) Estero La Tovara, Nayarit (localities 17-18), had genetic divergence from 0 to 0.04%. The species N. (N.) mamesi comprises 22 individuals from seven localities from Mexico, one of Guatemala and two from Costa Rica (localities 19-25 in Fig. 1), showing a genetic divergence from 0 to 1.3 %. N. (N.) golvani shows genetic divergence from 0 to 1.5% among 80 specimens from 11 localities (see localities 26-36). The species N. (N.) mexicoensis corresponding to 47 individuals from eight localities (localities 37-44) from Gulf of Mexico shows a genetic divergence ranged from 0 to 3%. N. (N.) schmidti shows low genetic divergence ranged from 0-0.02% of specimens analyzed from the same locality. The species N. (N.) emyditoides corresponding three localities: 1) Lago de Catemaco, Veracruz, 2) Río Papaloapan, Tlacotalpan, Veracruz and 3) Presa la Herradura, Monterrey (localities 46-48), showed a genetic divergence from 0 to1.3%. The specimens of N. (N.) panucensis are from Axtlan de Terrazas, San Luis Potosí, and Río Pantepec, Veracruz (localities 49-50), both localities are near of the type locality of N. (N.) panucensis . The genetic divergence among four isolates of N. (N.) panucensis ranged from 0.02–0.05%. Finally, the two specimens of N. (N.) chimalapasensis showed a genetic divergence equal to zero (locality 51).

1 2 3 4 5

1. N. (N.) brentnickoli (n = 81) 0–0.7 42–43 8.1–9.8 21.9–23.7 7.3–10 2. N. (N.) roseum (n = 2) 0–0.04 40.9–42.9 41.1–43.1 39.7–43.2 3. N. (N.) mamesi (n = 22) 0–1.3 21–23.6 2.40–5.50 4. N. (N.) golvani (n = 80) 0–1.5 21.0–24.6 5. N. (N.) mexicoensis (n = 47) 0–3.0 6. N. (N.) schmidti (n = 5)

7. N. (N.) emyditoides (n = 26)

8. N. (N.) panucensis (n = 4)

9. N. (N.) chimalapasensis (n = 2)

10. N. saginata (n =1)

continued.