Survey of Bioluminescent Coleoptera in the Atlantic Rain Forest of Serra da Paranapiacaba in São Paulo State (Brazil)

Brazil is the country with the largest number of bioluminescent beetle species in the world. However, estimates suggest that this number could much be higher, since many species remain to be discovered. In this work we made a survey of the species of bioluminescent beetles in Serra de Paranapiacaba – the largest remnant of Atlantic Forest in São Paulo State. The survey was done at Intervales State Park, Carlos Botelho State Park and municipality of Tapiraı́ and the following species were collected: Aspisoma lineatum, Aspisoma physonotum, Aspisoma fenestrata, Cratomorphus besckey, Cratomorphus distinctus, Photinus penai, Photinus sp1, Photinus sp9, Ethra aff. malledicta or axilaris, Ethra aff. adicta , Lucidotini incertae sedis, Cladodes flabellicornis, Cladodes demoulini, Amydetes lucernuta, Bicellonycha sp8, Bicellonycha ornaticollis, Pyrogaster lunifer, Pyrogaster moestus, Pyrogaster sp2, Pyrogaster sp5, Pyrogaster sp6, Photuris lugubris, Photuris sp1, Photuris sp7, Stenophrixotrix sp1, Brasilocerus sp1, Pseudophengodes sp1, Hapsodrilus pyrotis, Hypsiophtalmus sp1, Ptesimopsia luculenta, Pyroptesis cincticollis, Pyrearinus brevicolis, Pyrearinus candelarius, Pyrearinus micatus, Pyrophorus divergens. Our data show that Serra de Paranapiacaba is the second richest area in São Paulo state, especially in elaterids, with unique species typical of this area and species common to other investigated sites such as the Biological Station of Boracéia (in Salesópolis county) and the urbanized areas in the between Campinas SorocabaSão Paulo, originally covered to the Atlantic Rainforest.


Introduction
Brazil has the largest diversity of luminescent beetles in the world, about 500 described species, corresponding about 23% of described species in the world (Costa 2000;Viviani et al. 2010). These species are distributed mainly in the three main families of Elateroidea: Lampyridae, Phengodidae and Elateridae. Furthermore, two species of luminescent Staphylinidae were reported (Costa et al. 1986;Rosa 2010). These species are distributed mainly in the following biomes: Atlantic Rainforest, Cerrado (Savannah), Pantanal (Marshies) and Amazon Rainforest. Among these ecosystems, the Atlantic Rainforest is one of the richest and also most threatened ones, currently with only around 7-8% of the original cover or 11.4 -16% when considering the the small woods (Ribeiro et al. 2009).
Despite their biodiversity, taxonomic and systematic studies on bioluminescent beetles in Brazil are still scarce. Detailed modern taxonomical studies are found mainly for Elateridae by Costa e collaborators (Costa et al. 1988, Costa 1971a, 1971b1972, 1975. However, the families Lampyridae and Phengodidae lack recent reviews and their systematics remains troublesome. Some studies on the biology and ecology were performed for some species of Elateridae (Costa 1975, Costa et al. 1988) and Phengodidae (Costa et al. 1999, Viviani & Bechara 1997. In the family Lampyridae, bionomic studies were done for two semi-aquatic species of Aspisoma spp. (Costa et al. 1988, Viviani 1989, Aspisoma lineatum Gyllenhal, 1817 (Viviani et al., 2012), some lampyrid species of São Paulo State (Viviani 2001) and for Photuris fulvipes Blanchard 1837 (Rosa 2007). Recently, the genus Amydetes was revised with the description of several new species (Da Silveira and Mermudes, 2014).
Considering the growing importance of fireflies and bioluminescent beetles as potential environmental indicators (Viviani et al. 2010), mainly for the impact of artificial night-lighting (Lloyd 2006;Viviani et al., 2010;Hagen et al., 2014), and their importance for scientific studies and in biotechnology as source of bioluminescent reagents (Viviani 2007), it is urgent to make biodiversity surveys to aid conservation and bioprospection programs.
The Atlantic Forest is, after the Amazon, the second largest forest type in South America. The last corridor of this forest is located mostly in São Paulo state, being composed of a series of protected areas with more than 17.300 km 2 of forests (Galleti Jr et al. 2008). Considering the lack of knowledge about bioluminescent beetles in Brazil, in the nineties, and especially from 2006 we made surveys in the Atlantic rain forest. We already made lists of species occurring near the urban areas of Campinas, Rio Claro, Sorocaba and Votorantim cities in São Paulo State, which were mostly covered by seasonal Atlantic rain forest in the past (Viviani 2001, Viviani et al. 2010 and more recently about the bioluminescent species occurring in the hotspot of Biological Station of Boracéia located in the second largest remnant of Atlantic rain forest of Serra do Mar, along the coastline (Viviani & Santos 2012).
In this study we present a first survey made from 2007-2012 in the largest and most preserved remnant of Atlantic rain forest in São Paulo State, which is located along of Serra da Paranapiacaba, including three main investigated sites: (I) Cachoeira do Chá in the municipality of Tapiraí at its Northern border; (II) Parque Carlos Botelho in the middle and (III) Parque Intervales in the Southern part.

Materials and Methods
Habitats description. The investigated sites are located in a largest continuous remnant of Atlantic rain forest along the scarp chain Serra da Paranapiacaba, which is a subdomain of the Serra do Mar scarp chain, located about 100 km from the coast between the hydrographic basins: Sorocaba-Médio Tietê, Paranapanema and Ribeira-Southern Shore. The main kind of forest is dense mountain ombrophyl forest ( Fig.1) with the following phytophysionomic divisions: (I) mountain forest located over the mountains which include tall trees; (II) humid forest located between mountains over water bodies, which includes similar vegetation to the former habitat but with the presence of species better adapted to humid environments; (III) marshy areas at the borders of water bodies and streams which display typical grasses, and (IV) open fields surrounding the reservation or along thoroughfares opened in the forest. The following sites were investigated: Cachoeira do Chá, Tapiraí. (24°01'47.50"S -47°3429.47"W, 595m asl). This is a trail located along a stream at the Northern border of the large remnant, borthered by the route SP-79 and located 15 km from Tapiraí municipality. In this site the mountains display an escarpment ending at Ribeira River Valley.
Carlos Botelho. (24°03'46.48"S -47°59'30.31"W, 763m asl). This park features a rugged terrain in the upper level followed by escarpment ending at the Ribeira River Valley in the lower part ( Fig.1). Located in the north, at higher elevations there is a watershed of Ribeira River Valley Basin and another (Taquaral river) of Paranapanema River Basin. The mountains have dense rain forest (Montana forest), whereas in the lower part of the drive at the southeast, the vegetation is composed mainly by lowland tropical rain forest (Submontana forest). The survey sites within this unit are located near the headquarters, in the Northern part of the Park in the municipality of São Miguel Arcanjo.
Fazenda Intervales. (24°16'12.46"S -48°25'17.11"W, 826m asl). This site is located on the Serra da Paranapiacaba, and also on the Planalto de Guapiara, with very rough relief cut by headwater streams. The study site in this reserve was located between Guapiara and Ribeirão Grande municipalities. The present forest coverage is characterized as Montana forest as described above and the differential in this area is the geological and geomorphological formation, with many valleys and hills. The investigated areas were all located in the protected area, with the trails in the northwestern part of the Park. Adult lampyrids and elaterids were located by their own light at night and collected in flight with entomological net, or in the bushes or in the litterfall, during the period from August to April. They were also collected on the foliage during the day, especially diurnal lampyrids. Luminescent click-beetles and even some lampyrids such as Cratomorphus distinctus Olivier 1895 firefly could also be attracted to green chemiluminescent light sticks. Adult male phengodids were collected, in rare occasions, on soil and on the grass. Firefly larvae were collected on the undergrowth, in litterfall and soil at night, by the location of their glows (Viviani 2001, Viviani et al. 2010. Phengodid larvae were also collected at night in the soil, in embankments, and eventually on the underbrush by location of their luminescence (Viviani & Bechara 1997).
Identification. Specimens of fireflies and their larvae were identified first by comparison with scientific collection of Professor Viviani at UFSCar, which was previously identified by comparison with the collections of Museu de Zoologia da Universidade de São Paulo (São Paulo, Brazil), Natural History Museum of Paris (France) and British Museum (London,UK). Some species were identified by Dra. S. P. Rosa during her visit at the Natural History Museum of Paris (France). Elateridae species were previously identified by Dra. C. Costa (MZUSP) and Dra. S. P. Rosa. Several lampyrid and phengodid species could not be identified, and were catalogued by the name of the genus followed by a specific number (separated in morph species), according to Viviani & Bechara (1997) and Viviani (2001). The specimens were deposited in the collection in UFSCar under curatorship of V. Viviani.
The richness of each family of luminous beetles in each conservation unit is summarized in Figure 2. Lampyridae (Fig. 3). The number of observed lampyrid species was 10 in Carlos Botelho and 14 in the Intervales.
In Tapiraí only five species were found (see Table 1). The smaller number found in Tapiraí could be due to the lower frequency and smaller scanned area of collecting made in this site. The following species were unique to Carlos Botelho: Aspisoma lineatum, Photinus sp 1 , Ethra aff maledicta or axillaris, Lucidotini incertae sedis, Cladodes flabellicornis, Pyrogaster lunifer, Pyrogaster sp 5 and Pyrogaster sp 6 . Seven species were found at Intervales but were not been found in Carlos Botelho and Tapiraí: Aspisoma physonotum, Aspisoma fenestrata, Amydetes lucernuta, Bicellonychia ornaticollis, Pyrogaster moestus, Photuris sp 1 and Photuris sp 7 . There is a unique species which was collected only in Tapiraí -Photinus sp 9 -which displays a different bioluminescent signal from other Photinus species collected in Brazil, being characterized by a bimodal flash with bioluminescence spectrum peaking at 561 nm which is slightly blue-shifted in relation to other species of the genus found in the region (Viviani et al. 2010). Phengodidae.
Only four species of phengodids were found in Paranapiacaba. In Carlos Botelho only the arboreal Stenophrixotrix sp 1 larvae were found. In Tapiraí Brasilocerus sp 2 and Phrixotrix hirtus were found and in Intervales Pseudophengodes sp 1 . The arboreal Stenophrixotrix sp 1 was common to all three areas. When compared with Boraceia, the number of species and abundance of phengodids was lower.
The following species were found in both places: Brasilocerus sp2, Phrixotrix hirtus and Stenophrixotrix.
Elateridae (Fig. 4) Considering that the surveys were conducted in the same morphoclimatic domain and continuous geographic Mountain Tropical Rain Forest, it was surprising that several of these species were not found more widely distributed among these areas. This could be due to two reasons: (1) there is more heterogeneity in the habitats than assumed, and the same kind of specific habitats were not sampled in the three localities, or (2) because the survey was not conducted simultaneously in the three areas. In this case, the seasonality of adults of a given species may vary between areas.
The frequency of species per family in places where they are visibly more abundant is shown on Fig. 5. There is an apparent trend for greater richness for all families in Mountain Forest. More refined analysis with quantification of abundance is however required, to confirm whether the diversity of habitats is significant or not. Marshy areas seem to harbor a lower number of species.

Conclusions
The Serra de Paranapiacaba displays the second largest diversity of luminescent beetles in the Atlantic rain forest morphoclimatic domain of São Paulo state, with 34 species. Some species are common to sampled areas of semi-seasonal forest close to urbanized areas of inner São Paulo state and to the second largest and most preserved area of Biological Station of Boracéia. This area is especially rich in elaterid luminescent beetles, but displays poorer diversity of Phengodidae.