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This species occurs in cloud forest habitats, from 1400–2000 m elevation. All specimens are from Winkler or Berlese samples of sifted leaf litter. It is sympatric with R. isthmica at the type locality and with R. triumphalis on the slopes of Volcán Tacaná in Chiapas. At the type locality it was moderately abundant, occurring in 24 of 100 miniWinkler samples.
Diagnosis. Differing from R. isthmica in larger size (HW > 0.73 versus < 0.69), and with propodeal tooth larger and more acute.
Description. Worker. HW 0.73–0.83 (n=12); mandible with two or three teeth on masticatory margin (can vary within individuals, with two teeth on one mandible and three teeth on the other), second tooth from base largest; subapical tooth with distinct reclinate denticle at base; subapical tooth about twice as long as apical tooth; intercalary teeth prominent, one closest to apical tooth about half as long as apical tooth; labrum trapezoidal, anterior lobes triangular, inner margins of lobes shallowly sloping to semicircular median notch; metanotal groove moderately to strongly impressed; propodeal tooth usually large and acute (shorter and obtuse on one specimen), infradental lamella evenly and shallowly concave; squamiform setae abundant on first gastral tergite, either uniformly covering entire tergite or covering at least 3/4 of posterior portion. The queen and male are unknown.
Found most commonly in these habitats: 33 times found in cloud forest, 6 times found in cloud forest at night, 1 times found in 2º cloud forest, 1 times found in cloud forest litter, 1 times found in Liquidambar, oak, pine forest, 1 times found in oak, pine, Liquidambar forest.
Found most commonly in these microhabitats: 36 times ex sifted leaf litter, 5 times forest litter, 2 times leaf litter.
Collected most commonly using these methods: 24 times MiniWinkler, 6 times Berlese, 5 times MaxiWinkler, 6 times Night MiniWinkler, 1 times Winkler.
Elevations: collected from 1430 - 2000 meters, 1588 meters average
Type specimens: holotype Rhopalothrix megisthmica: casent0612564; paratype Rhopalothrix megisthmica: casent0603567, casent0603699, casent0612503, casent0614540, casent0614546, casent0629572, casent0629573