Thirty two new species ofLytopylus(Agathidinae) are described with image plates for each species:Lytopylus alejandromasisisp. n.,Lytopylus alfredomainierisp. n.,Lytopylus anamariamongeaesp. n.,Lytopylus angelagonzalezaesp. n.,Lytopylus cesarmoraisp. n.,Lytopylus eddysanchezisp. n.,Lytopylus eliethcantillanoaesp. n.,Lytopylus ericchapmanisp. n.,Lytopylus gahyunaesp. n.,Lytopylus gisukaesp. n.,Lytopylus guillermopereiraisp. n.,Lytopylus gustavoinduniisp. n.,Lytopylus hartmanguidoisp. n.,Lytopylus hernanbravoisp. n.,Lytopylus hokwonisp. n.,Lytopylus ivanniasandovalaesp. n.,Lytopylus johanvalerioisp. n.,Lytopylus josecortesisp. n.,Lytopylus luisgaritaisp. n.,Lytopylus mariamartachavarriaesp. n.,Lytopylus miguelviquezisp. n.,Lytopylus motohasegawaisp. n.,Lytopylus okchunaesp. n.,Lytopylus pablocobbisp. n.,Lytopylus robertofernandezisp. n.,Lytopylus rogerblancoisp. n.,Lytopylus salvadorlopezisp. n.,Lytopylus sangyeonisp. n.,Lytopylus sarahmeierottoaesp. n.,Lytopylus sergiobermudezisp. n.,Lytopylus sigifredomarinisp. n., andLytopylus youngcheaesp. n.A dichotomous key and a link to an electronic, interactive key are included. All specimens were reared from Lepidoptera larvae collected in Area de Conservación Guanacaste (ACG) and all are associated with ecological information including host caterpillar, collection date, eclosion date, caterpillar food plant, and locality. Neighbor-joining and maximum likelihood analyses of the barcode region of the mitochondrial cytochromecoxidase subunit I gene (COI DNA barcode) were conducted to aid in species delimitation.