A peer-reviewed open-access journal ZooKeys | 104: 29-54 (2022) AMO NREN IA #ZooKeys https:/ / ZOO keys. pensoft.net Launched to accelerate biodiversity research Life cycle and description of the immature stages of a terrestrial firefly endemic to Mexico: Photinus extensus Gorham (Coleoptera, Lampyridae) Martin L. Zurita-Garcia', Daniel Edwin Dominguez-Leén'”, Viridiana Vega-Badillo'”, Mireya Gonzdlez-Ramirez'”, Ishwari Giovanni Gutiérrez-Carranza', Geovanni M. Rodriguez-Mirén’, Sara Lépez-Pérez?, Paulina Cifuentes-Ruiz', Miriam Aquino-Romero', Santiago Zaragoza-Caballero! | Departamento de Zoologia, Instituto de Biologia, Universidad Nacional Auténoma de México, Apartado Postal 70-153, 04510, Mexico City, Mexico 2 Posgrado en Ciencias Biolégicas, Universidad Nacional Au- tonoma de México, Edificio D, 1° Piso. Circuito de Posgrados, Ciudad Universitaria, 04510, Mexico City, Mexico 3 Coleccién Coleopterolégica, Museo de Zoologia, Facultad de Estudios Superiores Zaragoza, Univer- sidad Nacional Auténoma de México, Av. Guelatao 66, Col. Ejército de Oriente, Alcaldia Iztapalapa, 09230, Mexico City, Mexico Corresponding author: Daniel Edwin Dominguez-Leon (biologiaunamedwin@gmail.com) Academic editor: Vinicius S. Ferreira | Received 16 January 2022 | Accepted 19 May 2022 | Published 7 June 2022 Attp://zoobank. org/68 CE42A 1-3B99-441 B-B15D-8E163C06A40B Citation: Zurita-Garcia ML, Dominguez-Leén DE, Vega-Badillo V, Gonzalez-Ramirez M, Gutiérrez-Carranza IG, Rodriguez-Mirén GM, Lépez-Pérez S, Cifuentes-Ruiz P, Aquino-Romero M, Zaragoza-Caballero S (2022) Life cycle and description of the immature stages of a terrestrial firefly endemic to Mexico: Photinus extensus Gorham (Coleoptera, Lampyridae). ZooKeys 1104: 29-54. https://doi.org/10.3897/zookeys.1104.80624 Abstract The life cycle, morphology, and bionomy of Photinus extensus Gorham, 1881, an endemic species of Mexico, are described. Redescriptions of adults (male and female) are also presented. Larvae were reared to the adult stage from eggs laid by females collected at the El Pedregal de San Angel Ecological Reserve, south of Mexico City. The activity period of adults of P extensus begins at the end of July and finishes by the end of August. Females lay between 3 and 198 eggs. Larvae hatch from the eggs after a period of 11 to 71 days, undergo 6 larval instars and a pupal stage in an annual cycle. Morphological characters of the sixth larval instar of P extensus are compared with those of several other genera assigned to the tribe Photinini. Knowledge of the natural history of firefly larvae is relevant since most species do not feed as adults and therefore depend on resources acquired during the larval stage. Keywords Bionomics, egg, larva, Photinini, pupa, redescription Copyright Martin L. Zurita-Garcia et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 30 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Introduction Fireflies belong to the family Lampyridae Rafinesque, 1815, and show a wide-ranging phenotypic and ecological diversity (Riley et al. 2021). Currently, there are more than 2400 described species with a worldwide distribution (Martin et al. 2019; Ferreira et al. 2020; Zaragoza-Caballero et al. 2020; Riley et al. 2021; Silveira et al. 2022). The highest species diversity is found in the Neotropical region (Costa 2000). Fireflies include nonluminous and luminous adults, luminous larvae and the females of some taxa are flightless (Branham 2010; Lewis et al. 2020). They inhabit wetlands, grass- lands, forests, agricultural fields and urban parks (Lewis et al. 2020). Many fireflies are strongly associated with particular habitats and vegetation types (Faust 2017). Firefly species can be either diurnally or nocturnally active. Diurnal species gener- ally do not have light organs as adults and rely on pheromonal and visual cues (Ohba 2004; Branham 2010). Luminous species are nocturnal or crepuscular, with biolumi- nescent signals produced from photic organs of various shapes and sizes located on abdominal ventrites. These visual signals are typically used in sexual signaling to com- municate species identity and facilitate pair formation (Branham 2010). Firefly larvae can be aquatic, semiaquatic or terrestrial and can be found along the mar- gins of streams and ponds as well as in leaf litter or rotten logs (Branham 2010). All known larvae are luminous and emit glows of varying duration. They are predatory on snails, earth- worms and other soft-bodied prey (Lewis et al. 2020). The adults of most species do not feed and therefore rely on resources gathered during the larval stages (Lloyd 2002; Vaz et al. 2020). Multiple genera endemic to the Neotropical region have no larval or pupal descrip- tions. The few species that have been studied have different life histories (Vaz et al. 2020). Currently, only the morphology of a small percentage of lampyrid larvae, at the generic or specific levels, of the approximately 144 genera and 2400 species, is known (Archangelsky 2010; Madruga and Branham 2020; Vaz et al. 2020; Zaragoza-Caballero et al. 2020; Riley et al. 2021; Silveira et al. 2022). There are a few studies describing the immature stages in the tribe Photinini. However, most of them have poorly detailed descriptions, composed only by the last larval stage (Riley et al. 2021). Besides, no tools to compare microstructures have been implemented. Of the 29 genera belonging to Photinini (Martin et al. 2019; Zaragoza-Caballero et al. 2020), there are detailed descriptions only for some species of Pyractonema Solier in Gay, 1849, Pyropyga, Mots- chulsky, 1852 Lucidina Gorham, 1883, Lucidota Laporte, 1833, Phosphaenus Laporte 1833 (Bugnion 1929; Beutel 1995; Branham and Archangelsky 2000; Archangelsky and Branham 2001; Archangelsky 2010, Kawashima 2017; Novak 2018b). Photinus Laporte, 1833 is the most diverse genus of the subfamily Lampyrinae with more than 300 described species (McDermott 1964; Zaragoza-Caballero et al. 2020). Members of this genus live in a variety of habitats (from tropical dry forests to temperate and tropical montane cloud forests). Species range from the United States to Argentina. One species, recently collected in Spain, was described as new (Zaragoza- Caballero and Vifiolas 2018) but then synonymized with the South American species Photinus signaticollis (Blanchard, 1846) (Koken et al. 2022). Life cycle and immatures of Photinus extensus oa In the past 20 years the number of known Photinus species has increased due to the description of new species from Mexico (Zaragoza-Caballero 2000, 2005, 2007, 2015, 2017; Zaragoza-Caballero et al. 2020). ‘This fact contrasts with the lack of knowledge of the larval stages and the natural history of these organisms. Until now, no study has accurately documented the life cycle duration of a Photinus species. This lack of knowl- edge is the result of the difficulty of collecting mated females, as well as challenges associated with rearing larvae under laboratory conditions. Moreover, some species are known to spend several years in the larval stage, and the time fireflies need to complete their life cycle depends on the geographical region and the availability of larval food (Buschman 2017). As for most genera in the family Lampyridae, Photinus larvae are poorly studied. Buschman (2017) observed that the first instars of this genus are smaller compared to those of Photuris Dejean, 1833, but no descriptions are provided. Most research docu- menting the natural history of Photinus focuses on the adult stage. These studies in- clude courtship in males (flash communication) (Wing 1991; Viviani 2001; Faust and Weston 2009; Faust 2010), nuptial gifts (Lewis et al. 2004a, 2004b), the utilization of ejaculate-derived proteins to nourish developing oocytes in certain species (Demary 2005) and the size of signal detection and emission organs (Lépez-Palafox et al. 2020). This paper documents the life cycle and larval morphology of a Photinus species for the first time. Photinus extensus Gorham, 1881 is an endemic species of Mexico. Its known distribution includes the state of Chiapas, Hidalgo, Mexico, Morelos and Mexico City (Zaragoza-Caballero et al. 2020). A reduced population of this firefly was also found in El Pedregal de San Angel Ecological Reserve, South of Mexico City. The life cycle of P extensus is herein presented, with descriptions of larval instar 6. Some characteristics of instars 1 and 3 are mentioned; the adult was described as well. Methods Collection Adults of P extensus were collected in the buffer zone of the El Pedregal de San Angel Ecological Reserve (19°19'28.82"N, 99°11'20.95"W); this zone is between the core zone and the urban area of Mexico City, it is totally in the territory of the Universidad Nacional Auténoma de México. El Pedregal de San Angel Ecological Reserve is lo- cated at the Southeast of Mexico City in the central Campus of the (UNAM) (Fig. 1). This community developed on a basaltic lava substrate approximately 1,670 years ago (Lot and Camarena 2009); the type of vegetation corresponds to a xerophytic shrub (Rzedowski 1978). The climate is sub-humid tempered with summer rains and an annual average precipitation of 833 mm; the annual average temperature is 15.5 °C (Orozco-Segovia et al. 2009). Fourteen adult females and 70 adult males of P exten- sus were collected on August the 3", 8", and 10° of 2018, between 19:30 and 21 h. Adult specimens were located by their bioluminescence in the undergrowth, where 32 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) 99°20'0"W 99°10'0"W 99°0'0"W 98°50'0"VW = fo) ran) rw) o re) — 19°30'0"N CONTOUR — 2201 msnm —— 2401 msnm —— 2601 msnm —— 2801 msnm —— 3001 msnm —— 3201 msnm —— 3401 msnm —— 3601 msnm 3801 msnm Kilometers 28 99°20'0"W 99°10'0"W 99°0'0"W 98°50'0"W Figure |. Map of Mexico City. Location of the study area where adults of P extensus were collected (indicated with a star). the dominant plant species is Pittocaulon praecox (Cav.) H. Rob. & Brettel (Asterales: Asteraceae). Other collections were made at the same site between June and July 2019, and five larvae were obtained. These developed into two female pupae and three males. Larvae, pupae, and adults were fixed in 70% ethanol for their preservation. Rearing in the laboratory To observe the reproductive activity of P extensus, adult specimens collected were divided into 14 groups consisting of five males and one female were placed in an 8 x 15 cm plastic container; peat moss substrate was added to simulate their natural en- vironment. After oviposition, eggs were placed over a gauze patch in a 50 x 100 mm Life cycle and immatures of Photinus extensus 33 Petri dish, and moistened every 48 hours with an antimycotic solution based on Ni- statine diluted in water (1/10). After eclosion, larvae werepartitioned into groups of five in separate Petri dishes (5x10 mm). To avoid dehydration, a filter paper layer was added and moistened every two days. This filter paper was replaced every week. Starting with the fourth larval instar, each larva was placed in a separate Petri dish (5x10 mm), with half of the dish covered with the filter paper, and the other half filled with sterilized dry sawdust. Following previous studies (Archangelsky and Bra- nham 2001; Archangelsky 2010), larvae were fed small pieces of earthworm Fisenia fetida (Savigny, 1826) (Haplotaxida: Lumbricidae). Dead or partially consumed prey were removed from the Petri dishes every two days to keep containers clean. Pupae were maintained in Petri dishes (5x10 mm) at room temperature with sterilized dry sawdust until they completed their development. Specimens representing the differ- ent larval instars, pupae, and adults, were preserved in 70% ethanol for reference and subsequent study. Morphological study Adults were identified using original descriptions, literature (Zaragoza-Caballero et al. 2000), and by comparison with photographs of type specimens (Natural His- tory Museum of London, BMNH) (Fig. 2A—F) and specimens identified by experts deposited at the Coleccién Nacional de Insectos (CNIN), Instituto de Biologia, UNAM, terminology of internal genitalia of females followed Silveira et al. (2022). Juvenile stages and adults of P extensus were examined under a Zeiss stereoscopic microscope (Discovery V8) with a 1x objective lens coupled with 16x eyepieces. Lar- val heads of each instar were separated from the body and immersed in 10% KOH solution, the mouthparts were dissected under a stereoscopic microscope and placed in glycerin on slides for observation. Description of the distribution of the setae was made for the last instar larva following Branham and Archangelsky (2000) and Arch- angelsky (2010). Pygopodial structure for the last instar larva was interpreted using Fu et al. (2012). Redescription of the adult of P extensus was made based on col- lected material. For the morphological description of larval instars, we followed the terminology of Novak (2018a) and Fu et al. (2012). A table 1 was made to compare morphological larval characters of Photinini (Pyractonema, Pyropyga, Lucidota, Lu- cidina, Phosphaenus and Photinus) (Branham and Archangelsky 2000; Archangelsky and Branham 2001; Archangelsky 2010; Novak 2018b), after Archangelsky (2010) and Kawashima (2017). Images were taken with an AxiocamMRC5 camera attached to a Zeiss Axio Zoom V16 microscope with an objective lens Plan NeoFluar Z, 1x10.25 FWD 56 at the Laboratorio de Microscopia y Fotografia de la Biodiversidad II, Instituto de Biologia, UNAM. Larvae were examined and imaged with a Hitachi SU1015 scanning electron microscope at the Laboratorio de Microscopia y Fotografia de la Biodiversidad I, Insti- tuto de Biologia, UNAM. 34 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) B.C.A.Col ITE, (2) ; B,C.A,Col.IIL. (2). ‘tty, Photinus 3 eine — Photinus : ) = ertlEnsud Moe NHMUK015011461 P MUKO15011462 Gortas O ii bem : i] . eaieoal) Sp. figured. Figure 2. Photinus extensus Gorham, type specimens NHM-London A general habitus of the adult male in dorsal view B ventral view C general habitus of the adult female in dorsal view D ventral view E male labels (lectotype) F female labels (paralectotype). Photos: Keita Matsumoto. Life cycle and immatures of Photinus extensus 35 Results Photinus extensus Gorham, 1881 Figs 3A—E, 4A-E [Adults] Redescription. Adult male (Fig. 3A, B) (n = 70). Length of body 16.1—20.3 mm; width 3.74.2 mm. Body brownish, except the pronotal disk with a central black spot and two red spots at the sides; protrochanters, coxae, seventh and eighth ventrites yel- lowish; light organ in sternites 5—6 and 7" sternite with diminished light spots. Head. \nterocular space flat, almost parallel, shagreen-like integument, brilliant and pilose; frons vertical, interantennal distance (0.17—0.22 mm; 0.2 + 0.01 mm) slightly wider than the antennal fossae (0.23—0.32 mm; 0.28 + 0.04 mm); eyes fine- ly faceted, semispherical, prominent, longer (1.1-1.18 mm; 1.17 = 0.03 mm) than wide (0.62—0.98 mm; 0.9 = 0.08 mm); antennae filiform, long (5.7—7.11 mm; 7.0 + 0.11 mm), one-and-a half times longer than pronotum, extending beyond the poste- rior coxae, scape claviform reaching a length of (0.69-0.98 mm; 0.79 = 0.10 mm), as long as the next two antennomeres together, the second short (0.19-0.79 mm; 0.28 + 0.05 mm), the third to the tenth (0.54—0.71 mm; 0.66 + 0.07 mm), the eleventh reaches (0.51—0.73 mm; 0.77 + 0.03 mm); frontoclypeal suture membranous, almost straight; clypeus trapezoidal, anterior margin concave, with setae along the margin; mandibles falcate, robust, with setae on the external base; maxillar palpomere ogival and robust, labial palpomere securiform. Thorax. Pronotum wider (3.95—4.4 mm; 4.04 + 0.08 mm) than long (2.8-3.4 mm; 3.01 + 0.27 mm), semicircular, with a longitudinal groove indistinct in the basal half, anterior margin rounded, posterior sinuate, posterior angles straight, sides narrowly ex- planate, with irregular glandular pores at the front and ordered on the posterior and lateral margins, surface brilliant, abundant pilosity, decumbent; scutellum spatulate, with the posterior margin rounded, surface brilliant, punctate and decumbent pilosity; long elytra, parallel, four and a half times longer (12—13.5 mm; 12.6 + 0.57 mm) than wide (2.4-2.8 mm; 2.58 + 0.19 mm), surface rugose, opaque, with two types of pilosity, one relatively long and erect, the other small and procumbent; mesothoracic respiratory spira- cles not tubular; long legs, pro, meso and metalegs similar to each other, femurs fusiform, tibiae channeled, a little dilated at the apex, external margin crenulate, two symmetric tibial spurs present in pro, meso and meta legs, tarsomeres laterally compressed, first met- atarsomere longer (0.6—-0.76 mm; 0.73 + 0.03 mm) than the next two metatarsomeres together (0.51—0.68 mm; 0.63 + 0.08 mm), fourth bifid, covering the fifth, claws simple. Abdomen. Sternites 5—6 longer than the preceding, with stigmatiform pores, poste- rior margin of sternite six cleaved, the seventh concave, the eighth ojival; posterior mar- gin of pygidium convex; aedeagus short, robust, with symmetrical basal piece (0.67— 0.7 mm; 0.68 + 0.05 mm), as long as lateral lobes (0.66—0.7 mm; 0.67 + 0.45 mm), with posterior margin concave, lateral lobes apically acute and convergent, median lobe cylindrical, with dorsal part membranous and ventral part with sclerosed base and api- cal half membranous, dorso-basal excrescences as long, oblique lobes, median orifice apical, lateral lobes narrowing towards the apex, apex blunt and wide (Fig. 3C, D, E). 36 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Figure 3. Photinus extensus Gorham A general habitus of the adult male in dorsal view B ventral view; Aedeagus: C dorsal view D lateral view E ventral view. Adult female (Fig. 4A, B) (n = 14). Length: 11.1-20.3 mm; width: 3.2-6.5 mm. Body brownish, except the pronotal disk with a central black spot and two red spots at the sides; procoxae, protrochanters, meso-coxae, meta-coxae, seventh and eighth sternites yellowish; light organ in fifth sternite. Life cycle and immatures of Photinus extensus 37 Head. \nterocular space flat, more or less parallel, shagreen integument, brilliant and pilose, frons vertical, interantennal distance (0.16—0.37 mm; 0.25 = 0.10 mm) wider than antennal fossae (0.16—0.37 mm; 0.44 + 0.16 mm) ; eyes small, finely facet- ed, semispherical, longer (0.65—0.94 mm; 0.67 = 0.25 mm) than wide (0.45—0.76 mm; 0.54 + 0.15 mm), antennae filiform, short (4.16—5.52 mm; 5.23 + 0.96 mm), as long as the length of pronotum, without extending beyond the posterior margin of metasternum; scape reaching a length of (0.57-0.64 mm; 0.58 + 0.34 mm), longer than the two next antennomeres together, the second short (0.2—-0.39 mm; 0.37 + 0.06 mm), from the third to the tenth (0.35—0.58 mm; 0.44 + 0.07 mm), the eleventh reaches (0.54—0.69 mm; 0.68 + 0.05 mm); frontoclypeal suture membranous, almost straight; clypeus trapezoidal, anterior margin concave, with setae along the margin; mandibles falcate, robust with setae on the external base; maxillar palpomere ogival and robust, labial palpomere securiform. Thorax. Pronotum wider (3.27—-4.97 mm; 4.21 + 0.86 mm) than long (2.21- 2.6 mm; 2.2 + 0.2 mm), semicircular, with a longitudinal groove indistinct on the basal half, anterior margin rounded, posterior margin straight, posterior angles straight, sides narrowly explanate, with glandular pores irregular at the front and ordered on the posterior and lateral margins, surface brilliant, pilosity abundant, decumbent, scutellum spatulate, with the posterior margin rounded, surface bril- liant, punctate and pilosity decumbent; elytra short, without covering the abdomen, two-and-a-half times longer (4.62—6.5 mm; 5.2 + 0.62 mm) than wide (1.91—2.71 mm; 2.22 + 0.42 mm), surface rugose, opaque, pilosity decumbent; divergent in the median margin, epipleura reduced, mesothoracic respiratory spiracles not tubular; legs similar to each other; tibiae and femurs flat, fusiform, tibiae channeled, a little dilated at the apex, external margin crenulate, tarsomeres laterally compressed, first metatarsomere (0.41—0.64 mm; 0.62 + 0.22 mm) slightly longer than the next two together (0.46—0.58 mm; 0.53 + 0.06 mm), the fourth bifid, covering part of the fifth, claws simple. Abdomen. Sternites 5—6 longer than the preceding, without stigmatiform pores, posterior margin of sternite six almost straight, the seventh cleaved, the eighth with a notched; posterior margin of pygidium convex. Internal genitalia with a short and rounded spermatophore-digesting gland, longer than spermatheca, bursa copulatrix with an elongated and weakly sclerotized plate. Ovipositor with valvifers free, two-and- a-half times longer (2.41—3.10 mm; 2.75 + 0.48 mm) than coxites (1-1.12 mm; 1.06 + 0.08 mm); coxites divergent posteriorly; styli minute, sclerotized; proctigerplate short with rounded posterior margin, well-sclerotized (Fig. 4C, D, E). Description of pre-imaginal stages. Egg (Fig. 5A, B). Semispherical shape, whitish, with a diameter of approximately 290-300 um (Fig. 5A). Surface with concavities that differ in size and shape (4— 10 um), evenly distributed, some present in aggregations (Fig. 5B). As time elapsed, the surface of the eggs became more transparent, allowing the observation of the larvae before hatching. 38 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Figure 4. Photinus extensus Gorham A general habitus of the adult female in dorsal view B ventral view C internal genitalia D ovipositor dorsal view E internal anatomy of the reproductive tract: spermatophore digesting (SDG) and spermateca (SPT). Life cycle and immatures of Photinus extensus 39 Figure 5. A Ege of Photinus extensus Gorham B surface with concavities that differ in size and shape (4-10 um), evenly distributed, some present in aggregations. Sixth instar larva (Figs 6A, B, 7A-H, 8A-G, 9A-F). Description. Elongate, ta- pering body, dorso-ventrally flattened, length 12.27—18.18 mm; integument of granu- lar appearance; tergites from protergum to abdominal segment IX divided by sagittal line in dorsal view. Tergites with two lateral pale stripes that run throughout the body to the VIII segment, more sclerotized than the sternites, with clearly visible setae on the posterior margin of tergites VII to X; the last tergum completely dark except the lateral margins paler; anterior margin of the first head segment with two fossae (sen- sorial or glandular) paler and bigger than the rest of the punctations of the segment. Membranous pleura except for a dark sclerotized area around the spiracles, without apparent setae. The ventral surface is flexible due to the intersegmental membranes. Mesothoracic and abdominal pleural areas of segments I—VIII with bilabiate spiracles. Head capsule. Prognathous; slightly visible when retracted into prothorax due to the transparency of the protergum; extensible neck membrane covered in extremely short spines forms a two-layer envelope around the head; partially retractable within the pro- thorax; completely sclerotized, small, wider (0.88—1.54 mm; 1.2+ 0.27 mm) than long (0.68-1.09 mm; 0.92 + 0.17 mm), flat, sides almost parallel; stemmata on each side, with an almost transparent spot located posteriorly to the stemmata; clypeus and labrum fused forming the clypeo-labrum covering base of the mandibles in dorsal view; maxillae and labium connate forming maxillolabial complex covering most of the ventral cephalic area; epicranial suture dark, U-shaped, with a very short epicranial stem, frontal arms V- shaped (Figs 7A, 8A). Epipharynx formed by two oval plates, without setae, that project centrally beyond the anterior margin of the head. Hypopharynx with short setation. Antenna. Irimerous, located on the distal margin of the epicranial plate; partially retractable into the antennal socket; three-segmented, basal antennomere and second antennomere (0.42—0.55 mm; 0.48 + 0.05 mm) elongated, and a third segment (the flagellum) short (0.24—0.31 mm; 0.26 + 0.03 mm); adjacent sensorial cone present; basal antennomere with two long setae in the anterior mid, almost entirely covered by moderately dense, second antennomere with long setae close to apex and entirely covered by dense smaller finer setae, third antennomere with long setae from base to apex, with short setae on the anterior margin (Fig. 7B, 8B). 40 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Figure 6. Photinus extensus Gorham A instar 6 in dorsal view B instar 6 in ventral view C instar 3 in dorsal view D instar 3 in ventral view E instar 1 in dorsal view F instar 1 in ventral view. Maxilla. Consisting of five parts, attached to lateral margins labium forming a maxillo-labial complex. Cardo elongate, irregular shape, with four setae in ventral sur- face, on long setae in posterior margin. Stipes elongated, ventrally covered with erect setae, with three long stout setae placed radially on the ventral apical region. Galea present, with two segments, the first longer and stouter than the apical, which is tri- angular (Fig. 7C, D). Lacinia covered with brush of long setae on outer lateral mar- gin. Maxillae with three-segmented palpi, basal segment long (0.57—0.71 mm; 0.66 + 0.06 mm) covered by setae in mid-region, segment II wider (0.30—0.40 mm; 0.35 + 0.04 mm) than long (0.12—0.16 mm; 0.14 + 0.01 mm); apical segment cylindri- cal (0.10—0.14 mm; 0.12 + 0.01 mm) with numerous setae from base to mid region; (Hies7 ERS aD): Life cycle and immatures of Photinus extensus 4] Labium. Closely attached to maxilla, formed by prementum, mentum and post- mentum. Prementum heart-shaped, surface covered with numerous short setae and two long setae close apex; labial palpi with two segments, basal palp subquadrate with few setae in mid-region, distal palp conical without setae; mentum with one pair of setae on anterior third and one pair of setae on posterior third; postmentum elongate, slightly sclerotized at the medial base, laterally united by membranes to the cardines; with a setae on each side near the base. Mandible. Symmetrical, falcate, strongly sclerotized, with an internal channel opening subapically on outer edge. Penicillus well-developed. Retinaculum short and rounded, present only as a blunt protuberance on basal third of the mandible. Densely covered by fine setae on the external margins basely, basal half on inner margin of mandible covered with a brush of stout setae, being longest on the retinaculous protu- berance (8E, F); mesal margin serrate. Thorax. Protergum wider (2.43-3.81 mm; 3.14 = 0.57 mm) than long (1.54— 2.59 mm; 2.11 + 0.43 mm), subsemicircular, wider posteriorly, rounded at poste- rolateral corners, covering the retracted head. Meso- and metatergum subrectangular three times wider than long, delimited by a pleural suture elongate barely evident from the laterotergites. Lateral areas of meso and metathorax scarcely sclerotized, composed of two laterotergites, the anterior with a well-developed spiracle on the mesothorax. Episterna extending from the anterior part to the lateral part of the coxae; epimeron forming a little sclerotized stripe, parallel to the coxae. Legs. Pentamerous, the first pair of forelegs slightly shorter than the second and third. Coxae short (0.76-1.16 mm; 0.98 = 0.18 mm), cylindric, widely separated at the base, decumbent; coxal-trochanteric membrane reaching about 1/3 of the coxal length. Trochanters pentagonal, joining the femur obliquely (0.51—0.82 mm; 0.71 + 0.13 mm). Femur narrow and cylindrical in lateral view. Tibiotarsus narrowing dis- tally with stout setae. All legs with a double row of long setae in the inner margin, numerous short setae in the outer margin: pretarsus claw-like with two setae at base (Figs 7G, H, 8G). Abdomen. Tergites IN-—IX of similar length (0.86—1.36 mm; 1.09 + 0.17 mm), width almost constant; segments I-VHI (or I-VII) wider than long, bearing a pair of long stout setae posterolaterally, with laterotergites at each side, with sclerotized plates containing the spiracles; ventral area of segments I—VIII with sternal areas almost squared, slightly pigmented, sternites with two long setae in mid-region; sternal medial area margined by laterosternites — sometimes pigmented, elongate, narrow, and paired, delimited by laterotergites dorsally, and ventrally by a medial sternal plate; ventral area of segment IX with a simple plate, without area differentiation; light organ present and segment VIII indistinct; abdomen ending with a series of eversible filaments (pygopo- dia) bifurcate at the apex, at least 30 pygopodia arise from 12 basal stalks which may branch more than once (the dorso and ventrolateral stalks branch into three); densely packed recurved hooks occur on the ventrolateral surface of each exerted pygopod and completely covering at apex, with toothed scales on the dorsolateral surface only on anterior half. (Fig. 9A—G). Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Figure 7. Sixth instar of Photinus extensus Gorham A SEM image of the head retracted and anterior part of prothorax in lateroanterior view B antennae C prementum, maxillary palpus and galea (lateroventral) D distal part of galea E, F maxillary palpi in ventrolateral view G right metathoracic leg in ventrolateral view H pretarsus. Life cycle and immatures of Photinus extensus 43 Third instar larva (Fig. 6C, D). Description. Similar to sixth instar. Length 11.45-13.78 mm. Head capsule. Wider (0.82-0.96 mm; 0.92+ 0.05 mm) than long (0.57—-0.85 mm; 0.71 + 0.12 mm). Antenna. Basal antennomere and second antennomere (0.28—-0.35 mm; 0.32 + 0.02 mm), and a third segment (the flagellum) (0.15-0.21 mm; 0.18 = 0.02 mm). Maxilla. Maxillae with three-segmented palpi, basal segment long (0.34—0.54 mm; 0.43 + 0.07 mm) covered by setae in mid region, segment II wider (0.30-0.43 mm; 0.36 + 0.06 mm) than long (0.12—0.17 mm; 0.14 + 0.02 mm); apical segment cylindrical (0.10—0.13 mm; 0.11 + 0.01 mm). Thorax. Protergum wider (1.36-1.53 mm; 1.4 + 0.06 mm) than long (1.18-1.68 mm; 1.3 + 0.20 mm), trapezoidal. Legs. Coxae short (0.77—0.97 mm; 0.86 + 0.08 mm), fe- mur obliquely (0.43—0.60 mm; 0.52 + 0.07 mm). Abdomen. Tergites III-IX (0.68- 0.85 mm; 0.77 + 0.045 mm) ventral area of segments I—VIII with sternal areas almost squared, slightly pigmented, sternites with two long setae in mid-region. First instar larva (Fig. 6E, F). Description. Similar to sixth instar. Length 3.72—8.80 mm; after hatching, the first instar body does not appear sclerotized to the degree found in later instars. Head capsule. Wider (0.48—0.72 mm; 0.64 0.10 mm) than long (0.24-0.51 mm; 0.38 + 0.12 mm), s (Figs 6A and 7A). AW. ANY ; Ay wey Figure 8. Photinus extensus Gorham, head structures of sixth instar larva A head capsule ventral view B right antenna, dorsal view C maxillary palpus and galea, ventral view D maxillary palpus, ventral view E mandible dorsal view F mandible ventral view and G leg dorsal view. 44 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Figure 9. Sixth instar of Photinus extensus Gorham A, B SEM image of pygopodium in general view and detailed C-F detail of the surface of pygopodium. Antenna. Basal antennomere and second antennomere (0.12—0.28 mm; 0.19 + 0.06 mm) elongated, and a third segment (the flagellum) short (0.08—0.17 mm; 0.12 = 0.03 mm). Maxilla. Maxillae with three-segmented palpi; basal segment long 0.24—0.47 mm; 0.31 + 0.08 mm) and well-defined, segment II wider (0.12- 0.17 mm; 0.15 = 0.01 mm) than long (0.05—0.07 mm; 0.06 = 0.007 mm); apical segment (0.03—0.047 mm; 0.037 + 0.007 mm). Labium. Postmentum elongate, slightly sclerotized at the medial base, laterally united by membranes to the car- dines; with a setae on each side near the base. Life cycle and immatures of Photinus extensus 45 Figure 10. Photinus extensus Gorham A pupa in dorsal view B pupa in ventral view. Thorax. Protergum wider (0.73—1.47 mm; 0.94 = 0.21 mm) than long (0.51- 0.75 mm; 0.53 + 0.09 mm). Legs. Coxae short (0.44—0.65 mm; 0.53 + 0.08 mm), femur obliquely (0.21—0.34 mm; 0.27 = 0.05 mm). Abdomen. Tergites III-IX (0.31- 1.2 mm; 0.74 + 0.28 mm). Pupa, male (Fig. 10A, B). Length 17-23 mm; width 6-7 mm. Body elongate, curved, ventrally concave, pale yellowish (sternites I-VHI slightly pigmented at the ends). Head. Totally covered by the pronotum in dorsal view. Large eyes, located at the sides of the head; antennae in front of the eyes, nearer the frontal center, mouthparts visible in ventral view. Thorax. Pronotum wider than long, semicircular, totally covering the head. Meso and metanotum shorter, subrectangular, bearing the elytra sidewards. All pairs of legs free, visible in ventral view. Spiracles present in the pleural areas of mesothorax. Abdomen. Abdominal segments subrectangular, wider than long, spiracles present on abdominal pleural areas of segments I-VIII. Light organ on sternites V—VI. Life cycle (Fig. 11). In their natural habitat, adults of P extensus are active from early July, when the first males can be observed. Bioluminescent activity begins at dusk, 46 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Egg 11-70 days Larva (six instar larva) Adult 10 days 286-323 days Pupa 7 days Figure I 1. Life cycle of Photinus extensus Gorham. at approximately 20:00 h, and diminishes considerably an hour later. Male flight does not exceed 2 m in height. Females are brachypterous and perch in the undergrowth approximately 50 cm from the ground. Males flash every 4.5 seconds, flying in an arc when illuminated. When males detect a female, they wait for an intense flash as a re- sponse, which is brief. The flash intervals are of 10 to 20 seconds. Males react by flying lower and towards the female. Groups of 3 to 5 males commonly compete with each other to get the female first, to mate with her. Two types of competition were observed among males: 1) a mating ball: four or more males cover the copulating pair and try to dislodge the copulating male to gain access to the female, and 2) males using their pronotum as a lever to pry a copulating male from the female. In the laboratory, copu- lation was observed to last from between 2 to more than 4 hours. During oviposition, females bend their abdomen and place the apical part of it on the substrate. Eggs are laid superficially or buried, randomly distributed, individually, Life cycle and immatures of Photinus extensus 47 or in groups (up to aggregates of 50). The number of eggs deposited by each female varied from three to 198. Eggs emit a faint bioluminescence since they are oviposited, which is only perceptible to the human eye in complete darkness. In total, 956 eggs from 13 females were obtained. Under lab condition P extensus completed its development in approximately 12 months, from oviposition to imago. The egg stage under laboratory conditions had a duration of 11 to 70 days, with mortality of n = 144 eggs (15%). Photinus extensus undergoes six similar larval instars that differ in both size and color (Fig. 4). Cannibalism among larvae during rearing was not observed. The only food larvae consumed was the earthworms provided. ‘There was no synchronization among larvae during the progression of larval instars, which started at the end of August until the beginning of July. In captive conditions, the process of ecdysis from one stage to the other varied among individuals. ‘The first larval instar had a duration of 14 to 153 days, where mortality was 60% (n = 491) among the eggs that hatched. The second larval instar had a duration of 14 to 172 days, with mortality of 61% (n = 199). The third larval instar had a duration of 15 to 140 days; mortality was 71% (n = 87). The fourth larval instar had a duration of 17 to 140 days, with a mortality of 52% (n = 20). The fifth larval instar had a duration of 24 to 192 days (n = 18) and the last larval instar had a duration of 53 days (n = 8). Pupation had a duration of 7 days in July, and according to the observations in the field, pupae were found under pyroclastic rocks. Discussion Photinus extensus has six instars; they are very similar and only differ in size, color and the degree of the body sclerotized and presence of setae. Instar III differs from I and VI by a trapezoidal pronotum (Fig. 5) and an exposed head. The periods of time of the instar I-V are variable in different specimens, ranging from 14 and 192 days; the last instar period in different specimens is constant, approximately 50 days; the pupa is completely developed in seven days. Frequently, the identification and description of larvae in the tribe Photinini is based on the characters present in the final larval instar, mostly body shape, color pat- tern, head capsule features, and the morphology of mouthparts (Archangelsky 2010). There are both similarities and differences between larvae belonging to the tribe Pho- tinini, including P extensus (see Table 1). The larval character suite found in P extensus is most similar to those found in Pyractonema, Lucidota, and Pyropyga. The major ex- ception to this is the number of segments in the maxillary palp. Although this last char- acter is shared with the genus Phosphaneus, it differs in the opening of the mandible channel. P extensus differs from Lucidina in the number of segments in the maxillary palp, and the number of retinacula of the mandible. There are some patterns among the larvae described in Photinini; the shape of the body is narrow and parallel and the pronotal shape is semicircular or semioval and the opening of the mandible channel is at the inner margin subapical. 48 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Table |. Morphological larval characters of known Photinini genera (after Archangelsky 2010, Kawashi- ma 2017 and Novak 2018b). Character | Pyractonema Pyropyga Lucidota atra Lucidina accensa Phosphaenus Photinus nigripennis | nigricans (Say) Olivier Gorham hemipterus (Goeze) extensus Solier Gorham Body shape __| Narrow, parallel | Narrow, parallel | Narrow, parallel | Wide, suboval 4.2 Oblong and narrow | Narrow, parallel 5.7-6.1x longer longer than narrow ~4,5x longer than ~5x longer than than narrow narrow narrow Ratio: body 3.43.7 3.43.7 3.14 29, length/thoracic length Cephalic Short and wide, | Short and wide, | Short and wide, |Subquadrate, moderately | Rectangular, retractable | Short and wide, capsule retractable into | retractable into | retractable into | flattened dorso-ventrally. into the thorax retractable into the thorax the thorax the thorax the thorax Antennae Wide, partially | Wide, partially | Wide, partially Long and thin, Long and thin, partially | Long and thin, retracted into | retracted into | retracted into the | completely retracted into] retracted into the head | partially retracted the head the head head the head into the head Opening of At the exterior | At the exterior | At the exterior | At the exterior margin, | At the exterior margin, | At the exterior the mandible margin, margin, margin, subapical subapical margin, channel subapical subapical subapical subapical Number of 1 retinacula of the mandible Maxillar palp Two palpomere Tew palpomere Four palpomere Shape of subcircular subcircular subcircular subcircular subcircular suboval pronotum Shape of sub oval sub oval sub oval trapezoidal sub oval rectangular mesonotum Shape of sub oval sub oval sub oval rectangular sub oval rectangular metanotum Thorax color Dark with Dark with Dark with Almost blackish dorssally dark reddish- | Dark, with pale three pale, three pale, three pale, brown, lateral and hind | brown, ventrally pink/ stripes longitudinal, | longitudinal | longitudinal and | margins more or less ochre/light brown and subparallel | and subparallel | subparallel lines | paler than the ground with darker plates lines lines Subrectangular on laterotergites and sternum Shape of the | Subrectangular, Suboval Suboval, Subrectangular I-VI, IX Subrectangular abdominal except VII-IX posterior margin subrectangular, VIII tergites subsquared of tergites V— suboval VII straight Abdomen With aclear | Dark with pale | Dark with three | Almost blackish brown, | pink/ochre/light brown | Pale with a little color line on each lateral areas _| pale longitudinal | lateral and hind margins pink stripe in the side. Dark segments VI-IX. (segments I— VID) and subparallel lines, at the inner interior margin more or lesspaler than the ground I-VI, segments VII-X pale yellowish to milky white second third in segments IV, the rest of them pale The distribution of the setae is different between genera (Table 2), chaetotaxy will be useful in Lampyridae as a tool to distinguish between immatures, also as a source of informative phylogenetics characters (Ballantyne et al. 2019; Riley et al. 2021; Vaz et al. 2021). However, the few studies on the morphology of immatures in Lampyri- dae and the lack of knowledge of chaetotaxy hinder comparison between fireflies and makes the elaboration of more detailed hypotheses very difficult. Life cycle and immatures of Photinus extensus Table 2. Chaetotaxy larval characters. Photinus extensus Gorham Pyractonema nigripennis Solier 49 Lucidota atra Olivier Hypopharynx Margin covered by dense pubescence. Surface covered by dense No information. pubescence. Mandible With patches of dense pubescence in the With patches of dense Mid-region covered by only one row basal part in ventral Margin covered by dense pubescence. With patches of dense pubescence in the basal part in ventral view. Mid-region covered by a single row of long setae. pubescence in ventral view. Mid-region covered by only one row of setae. of long setae. One long seta close to the apex. With 13-15 setae in ventral surface. Basal palp covered by setae. Palp II with some setae. Last palp without setae. Without setae. Basal palp with long setae from the mid region to the apex. Palp II with shorter setae. Distal palp without setae. Basal palp with some setae. Distal palp without setae. Dorsal and ventral surface with many setae. Basal palp with few long setae. Distal palp with one long seta. With two basal regions of very fine setae, with longer setae on the palp segment. With two long setae in the basal middle. No information. All antennomeres covered by setae. Antennomere III with many short setae. With a double row of long setae in the inner margin. Pretarsus with two setae at the base. Basal antennomer with mid region covered by setae. Anterior region with longer setae. Antennomere II covered evenly by long setae. Antenomere III with short setae. With a double row of long setae in the inner margin. Pretarsus with two setae at the base. Cardo With 4 long setae in ventral surface and one long seta in the posterior margin. Maxillary Basal palp covered by setae in the mid region. palpomeres _| Distal palp with setae from base to mid region. Labial Basal palp with few setae in mid region. Distal palpomeres palp without setae. Prementum Surface with many setae. Two long setae close to apex. Submentum With two long setae in mid region. Antennomeres | Apex with two long setae. Antennomere II] with long setae from base to apex. Legs With a double row of long setae in the inner margin. Outer margin with many setae. Pretarsus with two setae at the base. Abdomen Sternites with two long setae in mid-region. Sternites with two long setae in mid-region. No information. The larval characters have shown to be important to clarify the phylogenetic rela- tionships as Archangelsky (2010) mentioned. However, it seems to be a difficult task to get a good dataset of larval characters of Lampyridae. The life cycle, morphology, and behavior of the species of Photinus are similar. Photinus carolinus Green, 1956, P ignitus Fall, 1927, P marginellus LeConte, 1852, P pyralis (Linnaeus, 1767), P greeni Lloyd, 1969, and P extensus are the known species that produce spermatophores due to the prolonged time of copulation ( Cratsley et al. 2003; South and Lewis 2012). The length of the pupal stage varies slightly in Photinus. The pupal stage of P extensus and P carolinus has a duration of six days (Faust 2010). Nevertheless, the pupa of P caro- linus is present during May, while that of P extensus is present in July. Another difference is that the pupae of P extensus observed in the field were found under pyroclastic rocks, in contrast with P carolinus, which has been reported to occur under leaf-litter, near rot- ten logs, or moss (Faust 2010). Until now, little information has come to light about the pupae in other genera of Photinini (Archangelsky and Branham 2001). In other genera like Aspisoma Laporte, 1833, of Cratomorphini, the pupal stage is similar, occurring during a short period between six and ten days (Costa et al. 1988; Archangelsky 2004). 50 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Conclusion The complete life cycle of Photinus extensus, including descriptions of egg, larvae, and pupa, was documented for the first time. Larvae were reared in laboratory conditions to the adult stage from eggs. The six instar of P extensus are very similar; they differ only in size and in the sclerotized degree. Among Photinini larvae there are not many differences, differing in the number of segments in the maxillary palp and in the number of retinacula of the mandible; the shape of the body and pronotum, and the opening of the mandible channel fol- low a similar pattern. Life cycle information is essential to carry out protection and conservation actions for insects that are very sensitive to environmental changes, like fireflies. For example, the species that do not produce light are easily overlooked and the information about their life cycle is deficient. This results in “Data Deficient” cat- egorization in evaluations of extinction risk. Thus, more studies are needed in which the life history, habitat associations, and microhabitat are detailed (Fallon et al. 2021). Also, it is necessary to understand that requirements of larvae are different to those of adults to have an integral vision in the actions of protection of the fireflies. Acknowledgements We thank Susana Guzman (IBUNAM) for help with the use of the Zeiss equipment, and Berenit Mendoza (IBUNAM) for the electron micrographs. We are grateful to Marc A. Branham and Sarah Sander Lower for their work correcting the English text in the manuscript and their valuable comments. We also thank Simone Policena Rosa, Luiz Silveira and Oliver Keller for helping to improve the manuscript. We thank the Natural History Museum of London, Michael Geiser and Keita Matsumoto for pro- viding photographs of the lectotype. MLZG, DEDL, PCR and IGGC field work; IGGC and DEDL designed the breeding method; DEDL, IGGC, MGR and VVB laboratory work; SZC and IGGC identified the species; VVB, MGR, GMRM and MAR produced the photographs. SLP and DEDL constructed tables 1 and 2. All au- thors wrote the manuscript, revised it, and discussed the data. References Archangelsky M (2004) Description of the last larval instar and pupa of Aspisoma fenestrata Blanchard, 1837 (Coleoptera: Lampyridae) with brief notes on its biology. Tijdschrift voor Entomologie 147(1): 49-55. https://doi.org/10.1163/22119434-900000138 Archangelsky M (2010) Larval and pupal morphology of Pyractonema nigripennis Solier (Co- leoptera: Lampyridae: Photinini) and comparative notes with other Photinini larvae. Zootaxa 2601(1): 37-44. https://doi.org/10.11646/zootaxa.2601.1.3 Life cycle and immatures of Photinus extensus 51 Archangelsky M, Branham MA (2001) Description of last instar and pupa of Pyropyga nigri- cans (Coleoptera: Lampyridae, Photinini) and comparison with larvae of other Photinini genera. Canadian Entomologist 133(2): 155-164. https://doi.org/10.4039/Ent133155-2 Ballantyne LA, Lambkin CL, Ho J-Z, Jusoh WFA, Nada B, Nak-Eiam S, Thancharoen A, Wattana- chaiyingcharoen W, Yiu V (2019) The Luciolinae of S. E. Asia and the Australopacific region: a revisionary checklist (Coleoptera: Lampyridae) including description of three new genera and 13 new species. Zootaxa 4687(1): 1174. https://doi.org/10.11646/zootaxa.4687.1.1 Beutel RG (1995) Phylogenetic analysis of Elateriformia (Coleoptera: Polyphaga) based on larval characters. Journal of Zoological Systematics and Evolutionary Research 33(3—4): 145-171. https://doi.org/10.1111/j.1439-0469.1995.tb00969.x Branham MA (2010) Lampyridae Latreille, 1817. In: Leschen RAB, Beutel GR, Lawrence JF (Eds) Coleoptera, Beetles. Volume 2: Morphology and Systematics (Elateroidea, Bostrichi- formia, Cucujiformia partim). Walter de Gruyter, Berlin, Germany, 141-149. https://doi. org/10.1515/9783110911213.141 Branham MA, Archangelsky M (2000) Description of the last larval instar and pupa of Lucidota atra (G.A.Olivier, 1790) (Coleoptera: Lampyridae), with a discussion of abdominal seg- ment homology across life stages. Proceedings of the Entomological Society of Washington 102(4): 869-877. Bugnion E (1929) Le ver-luisant provencal et la Luciole nicoise. Association des Naturalistes de Nice et des Alpes-Maritimes; Memoire Supplement au “Riviera Scientifique’, 131 pp. Buschman LL (2017) Analysis of courtship flash behavoiur in two Photuris fireflies (Coleoptera: Lampyridae) with field validation and rearing notes. In: Day JC (Ed.) Lampyrid: The Jour- nal of Bioluminescent Beetle Research. Abingdon, CreateSpace Independent Publishing Platform, 1-19. Costa C (2000) Estado de conocimiento de los Coleoptera neotropicales. In: Martin-Piera F, Morrone JJ, Melic A (Eds) Hacia un proyecto CYTED para el inventario y estimacion de la diversidad entomolégica en iberoamérica: Pribes 2000. Sociedad Entomoldgica Aragonesa, Zaragoza, 99-114. Costa C, Vanin SA, Casari-Chen SA (1988) Larvas de Coleoptera do Brasil. Museu de Zo- ologia, Universidad de Sao Paulo, Brazil, 282 pp. https://doi.org/10.5962/bh1. title. 100233 Cratsley CK, Rooney JA, Lewis SM (2003) Limits to nuptial gift production by male fireflies, Photinus ignitus. Journal of Insect Behavior 16(3): 361-370. https://doi. org/10.1023/A:1024876009281 Demary K (2005) Sperm storage and viability in Photinus fireflies. The Journal of Physiology 7: 837-841. https://doi.org/10.1016/j.jinsphys.2005.04.001 Fallon CE, Walker AC, Lewis S, Cicero J, Faust L, Heckscher CM, Pérez-Hernandez CX, Pfeiffer B, Jepsen S (2021) Evaluating firefly extinction risk: Initial red list assess- ments for North America. PLoS ONE 16(11): 1-18. https://doi.org/10.1371/journal. pone.0259379 Faust LF (2010) Natural history and flash repertoire of the synchronous firefly Photinus caroli- nus (Coleoptera: Lampyridae) in the great Smoky Mountains National Park. The Florida Entomologist 93(2): 208-217. https://doi.org/10.1653/024.093.0210 52 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Faust LF (2017) Fireflies, glow-worms, and lightning bugs: identification and natural history of the fireflies of the Eastern and Central United States and Canada. Athens (GA), University of Georgia Press, USA, 356 pp. Faust LE, Weston PA (2009) Degree-day prediction of adult emergence of Photinus carolinus (Coleoptera: Lampyridae). Environmental Entomology 38(5): 1505-1512. https://doi. org/10.1603/022.038.0519 Ferreira VS, Keller O, Branham MA (2020) Multilocus phylogeny support the nonbiolu- minescent firefly Chespirito as a new subfamily in the Lampyridae (Coleoptera: Elat- eroidea). Insect Systematics and Diversity 4(6): 1-13. https://doi.org/10.1093/isd/ ixaa014 Fu X, Ballantyne LA, Lambkin CL (2012) The external larval morphology of aquatic and ter- restrial Luciolinae fireflies (Coleoptera: Lampyridae). Zootaxa 3405(1): 1-34. https://doi. org/10.11646/zootaxa.3405.1.1 Kawashima I (2017) Larval morphology of the lampyrine species, Lucidina accensa Gorham (Coleoptera: Lampyridae: Lampyrinae) from Honsht, Japan. Japanese journal of system- atic entomology = HA BL Ray RF SK, 23(1): 129-134. Koken M, Guzman-Alvarez JR, Gil-Tapetado D, Romo Bedate MA, Laurent G, Rubio L, Ro- vira Comas S, Wolffler N, Verfaillie EK De Cock R (2022) Quick spreading of populations of an exotic firefly throughout Spain and their recent arrival in the french Pyrenees. Insects 13(2): e148. https://doi.org/10.3390/insects 13020148 Lewis SM, Cratsley CK, Demary K (2004a) Mate recognition and choice in Photinus fireflies. Annales Zoologici Fennici 41: 809-821. Lewis SM, Crastley CK, Rooney JA (2004b) Nuptial Gifts and Sexual Selection in Photinus Fireflies. Integrative and Comparative Biology 44(3): 234-237. https://doi.org/10.1093/ icb/44.3.234 Lewis SM, Wong CH, Owens A, Fallon C, Jepsen S, Thancharoen A, Wu C, De Cock R, Novak M, Lépez-Palafox T, Khoo V, Reed JM (2020) A Global Perspective on Firefly Extinction Threats. Bioscience 70(2): 157-167. https://doi.org/10.1093/biosci/biz157 Lloyd JE (2002) Family 62. Lampyridae. In: Arnett Jr RH, Thomas MC, Skelley PE, Frank JH (Eds) American Beetles vol. 2: Polyphaga: Scarabaeoidea through Curculionoidea. CRC Press, Boca Raton, Florida, 187-196. Lépez-Palafox T, Macias-Ordofez R, Cordero CR (2020) ‘The size of signal detection and emis- sion organs in a synchronous firefly: Sexual dimorphism, allometry and assortative mating. Peer] 8: €10127. https://doi.org/10.7717/peerj.10127 Lot A, Camarena P (2009) El Pedregal de San Angel de la ciudad de México: reserva ecoldgica urbana de la Universidad Nacional. In: Lot A, Cano-Santana Z (Eds) Biodiversidad del ecosistema del Pedregal de San Angel. Libro conmemorativo del 25, 19-25. Madruga O, Branham MA (2020) Description of life cycle and preimaginal stages of Alecton discoidalis Laporte, 1833 (Coleoptera: Lampyridae) under laboratory conditions. Zootaxa 4816(1): 81-91. https://doi.org/10.11646/zootaxa.4816.1.4 Martin GJ, Stanger-Hall KE Branham MA, Da Silveira LFL, Lower SE, Hall DW, Li XY, Lemmon AR, Lemmon EM, Bybee SM (2019) Higher-Level Phylogeny and Reclassifica- tion of Lampyridae (Coleoptera: Elateroidea). Insect Systematics and Diversity 3(6): 1-15. https://doi.org/10.1093/isd/ixz024 Life cycle and immatures of Photinus extensus a0 McDermott FA (1964) The taxonomy of the Lampyridae. Transactions of the American Ento- mological Society 90: 1-72. Novak M (2018a) Redescription of immature stages of central European fireflies, Part 2: Lamprohiza splendidula (Linnaeus, 1767) larva, pupa and notes on its life cycle and be- haviour (Coleoptera: Lampyridae). Zootaxa 4378(4): 516-532. https://doi.org/10.11646/ zootaxa.4378.4.4 Novak M (2018b) Redescription of immature stages of central European fireflies, Part 3: Phosphaenus emipterus (Goeze, 1777) larva, pupa and notes on its life cycle and behavior, with a key to three Central European lampyrid larvae (Coleoptera: Lampyridae). Zootaxa 4382(3): 450-464. https://doi.org/10.11646/zootaxa.4382.3.2 Ohba N (2004) Flash communication systems of Japanese fireflies. Integrative and Compara- tive Biology 44(3): 225-233. https://doi.org/10.1093/icb/44.3.225 Orozco-Segovia A, Gamboa de Buen A, Barrandas-Miranda VL (2009) La diversidad funcional de los ecosistemas. In: Lot A, Cano-Zantana Z (Eds) Biodiversidad del Ecosistema en el Pedregal del San Angel. Universidad Nacional Auténoma de México, México, 295-316. Riley WB, Rosa SP, Lima da Silveira LF (2021) A comprehensive review and call for studies on firefly larvae. Peer] 9: 1-24. https://doi.org/10.7717/peerj.12121 Rzedowski J (1978) Vegetacion de México, Limusa, México, 247-273. Silveira LFLd, Lima W, Fonseca CRVd, McHugh J (2022) Haplocauda, a new genus of fireflies endemic to the Amazon rainforest (Coleoptera: Lampyridae). Insects 13(1): e58. https:// doi.org/10.3390/insects13010058 South A, Lewis SM (2012) Effects of male ejaculate on female reproductive output and lon- gevity in Photinus fireflies. Canadian Journal of Zoology 90(5): 677-681. https://doi. org/10.1139/22012-031 Vaz S, Lima da Silveira LE Policena RS (2020) Morphology and life cycle of a new species of Psilocladus Blanchard, 1846 (Coleoptera, Lampyridae, Psilocladinae), the first known bromeliad-inhabiting firefly. Papéis Avulsos de Zoologia 24(special issue): 1-15. https:// doi.org/10.11606/1807-0205/2020.60.special-issue.24 Vaz SNC, Guerrazzi MC, Rocha M, Faust LE, Gabriel Khattar G, Mermudes JRM, Silveira LFL (2021) On the intertidal firefly genus Micronaspis Green,1948, with a new species and a phylogeny of Cratomorphini based on adult and larval traits (Coleoptera: Lampyri- dae). Zoologischer Anzeiger 292: 64-91. https://doi.org/10.1016/j.jcz.2021.01.002 Viviani VR (2001) Fireflies (Coleoptera: Lampyridae) from Southeastern Brazil: habitats, life history, and bioluminescence. Annals of the Entomological Society of America 94(1): 129- 145. https://doi.org/10.1603/0013-8746(2001)094[0129:FCLFSB]2.0.CO;2 Wing SR (1991) Timing of Photinus collustrans reproductive activity: finding a mate in time (Coleoptera: Lampyridae). Coleopterists Bulletin 45: 57-74. Zaragoza-Caballero S (2000) Cantharoidea (Coleoptera) de México. IV. Nuevos Photinus (Lampyridae) del estado de Morelos. Dugesiana 7(1): 1-17. Zaragoza-Caballero S (2005) Nuevas especies de Photinus (Coleoptera: Lampyridae: Photinini) de Jalisco, México. Folia Entomologica Mexicana 44(1): 75-82. Zaragoza-Caballero S (2007) A new species of Photinus (Coleoptera: Lampyridae: Photinini) from Jalisco, Mexico, with comments on intraspecific aedeagal variability and a key to the species of the subgenus Paraphotinus. Zootaxa 1437(1): 61-67. 54 Martin L. Zurita-Garcia et al. / ZooKeys 1104: 29-54 (2022) Zaragoza-Caballero S (2015) Nuevas especies de Photinus (Coleoptera: Lampyridae: Photinini) del bosque tropical caducifolio de Pacifico mexicano. Revista Mexicana de Biodiversidad 86(3): 638-65 1. https://doi.org/10.1016/j.rmb.2015.08.001 Zaragoza-Caballero S (2017) Nuevos Photinus Laporte, 1832 (Coleoptera: Lampyridae: Photinini). Dugesiana 24(2): 221-230. https://doi.org/10.32870/dugesiana.v28i2.7161 Zaragoza-Caballero S, Viholas A (2018) Photinus immigrans sp. nov. (Coleoptera: Lampyridae: Photinini): Primer registro del género Photinus en Catalufia, Espana. Revista gaditana de Entomologia 9(1): 273-286. Zaragoza-Caballero S, Lopez-Pérez S, Vega-Badillo V, Dominguez-Leén DE, Rodriguez-Mirén GM, Gonzdlez-Ramirez M, Gutiérrez-Carranza IG, Cifuentes-Ruiz P, Zurita-Garcia ML (2020) Luciérnagas del centro de México (Coleoptera: Lampyridae): descripcién de 37 es- pecies nuevas. Revista Mexicana de Biodiversidad 91(0): 1-70. https://doi.org/10.22201/ ib.20078706e.2020.91.3104