**1. Introduction**

46 Zoology

Yamashita, N.; Stoner, K.E.; Riba-Hernández, P.; Dominy, N.J. & Lucas, P.W. (2005). Light

1045-2249

levels used during feeding by primate species with different colour vision phenotypes. *Behavioural Ecology and Sociobiology*, Vol.58, No.6, pp. 618-629, ISSN

> Species diversity does have a pivotal role in the study and perception of biodiversity (Boero, 2010). One of the goals of Zoology is the study of the animal diversity, that is, the animal species, and Taxonomy is one of the basic disciplines to achieve it. Currently, biodiversity research requires a multidisciplinary approach (Boero, 2009), many different disciplines being involved, such as morphology, molecular biology, ecology, ethology…, that provide new characteristics to be considered. The study of biodiversity should proceed with the contribution of integrative taxonomy (Boero, 2010), taking into account, in addition to the traditional taxonomy, other disciplines of great utility, such as the study of the behaviour.

> In this context, behaviour and sounds are relevant characteristics to discover new taxa (Valdecasas, 2011). Sound production in insects is widespread and has been recorded in different orders. It is involved in different behaviours, the most important of which are defence against predators, aggression and mating or sexual behaviour. Four types of senses are used by insects in their sexual behaviour: tactile, visual, chemosensory and acoustics. The acoustic sense is that generally have received more attention, this stimulus is often heard by humans and their production involves the movement of specialized structures that can be seen, usually directly. The types of sounds produced and producing mechanisms can be framed primarily in three categories: stridulation, vibration and percussion. The sounds of insects have been classified in the context of behaviour in several types: call or proclamation, courtship, aggregation, aggression, mating and sounds of interaction (Lewis, 1984; Ragge & Reynolds, 1998).

The Acoustic Behaviour as a Tool for Biodiversity and Phylogenetic Studies: Case of

et al., 2003; Lorier, 1996; Lorier et al., 2010; Riede, 1987).

Lavalleja Departments (Assis-Pujol, 1997a, 1998).

Uruguay, and specimens captured in Uruguay.

Measurements are expressed in mm (Table 2).

**2. Materials and methods** 

coated with pure gold.

the *Rhammatocerus* Species Inhabiting Uruguay (Orthoptera, Acrididae, Gomphocerinae) 49

Neotropical Gomphocerinae has been studied to a very limited extent (COPR, 1982; García

*Rhammatocerus pictus* (Brunner, 1900) and *Rhammatocerus brunneri* (Giglio-Tos, 1895) are robust species inhabiting wet high pastures in northern and central South America, between about latitudes 15 and 40 South. *Rhammatocerus pictus* is distributed in central and southern Brazil (Rio Grande do Sul), Paraguay, Chile (Malleco), northern and central Argentina, Bolivia and Uruguay (Assis-Pujol, 1998; Carbonell et al., 2006) and *Rhammatocerus brunneri* in central-southern Brazil, Paraguay, Bolivia and Uruguay (Assis-Pujol, 1997a). In Uruguay, both species occupy similar habitats (low humid areas covered with tall and dense grass and low hill slopes). They are distributed in the country (Assis-Pujol, 1998) and are more frequently found north of Río Negro. Both species are sympatric in Artigas, Rivera and

These two species are systematically close to each other, having been separated by Assis– Pujol (1998) just on the basis of morphological criteria: colour of hind femora and tibiae and

Our objective is to establish the real status of these two taxa at the light of their sound production and acoustic behaviour, describing the songs produced in different behavioural

This study on the sound production and stridulatory structures of *Rhammatocerus pictus* and *Rhammatocerus brunneri* was conducted with males and females proceeding from the "Colección de Entomología de la Facultad de Ciencias de la Universidad de la República",

To study the structure of the stridulatory apparatus, 10 specimens of each sex and species (Table 1) were observed under a binocular microscope (Olympus SZH provided with 10X ocular lenses, 0.66-4X zoom objective, 2X lens and graduated eye piece) as well as with a Jeol 6100 scanning microscope, equipped with SEI (secondary electron images), working at an acceleration voltage of 15 kV and at 21 mm (working distance). Images were captured with the LINK ISIS program. Because the samples are hard and have not risk of being dehydrated, they only had to be cleaned as proposed by Clemente et al. (1989) and, then,

Measurements were taken by means of a sliding stage mounted on the stereoscopic binocular microscope, the displacement of which was measured by an attached dial calliper in combination with a graduated eye piece. The accuracy of the dial calliper was 0.05 mm.

For this study, the shape of the file and pegs have been taken into account, as well as the number of pegs in the file, femur length (HFL), file length (FL), peg density all along the file (PD) and in its middle area (PDM) and file length / femur length ratio (FLx100/HFL).

situations, the behavioural units identified and the sound-morphological structures.

**2.1 Microscopy techniques and characters considered in stridulatory file study** 

the spermatheca shape. Their acoustic behaviour is up to date unknown.

The sound on Orthoptera serves to promote social relations in the broadest sense of the term. This type of acoustic behaviour cannot be studied in isolation and cannot be understood except within the framework of the general behaviour of the species that is not just the sound production. From this point of view, we show that an exact knowledge of the physical elements of the sounds, as well as the morphology of these sound-producing organs is essential (Busnel, 1954). The sounds produced by the Orthoptera are important from a taxonomic point of view and has an important role as a mechanism of identification. They are of great value to establish the real status of local populations that may have few morphological differences (Blondheim, 1990; García et al., 1995).

The process of acoustic communication in reproduction is amply documented in Ensiferan insects (crickets and katydids): males of singing Ensifera (Orthoptera) emit specific calling songs used for species recognition, while courtship songs are generally less specific and could be mostly under the influence of sexual selection. In contrast, singing Caelifera (Orthoptera) perform more diverse behaviour prior to mating. In the subfamily Gomphocerinae (Acrididae) more specifically, many species emit calling songs, which are sufficiently specific to be used for species identification in the field, but they also perform very complex, often multimodal, courtship behaviour involving sequences of acoustic, vibrational and/or visual signals (Nattier et al., 2011; Ragge & Reynolds, 1998).

Neotropical Gomphocerinae form a group of grasshoppers whose taxonomy, systematics and biology are still poorly known (Otte, 1979; Otte & Jago, 1979). Within this group, *Rhammatocerus* Saussure, 1861 is widely distributed, from southern USA to central Argentine. Its species have a great economic importance; many of them are important crop and grazing pests (Carbonell et al., 2006; Cigliano & Lange, 1998; Salto et al., 2003), especially in Brazil and Colombia (Lecoq & Assis-Pujol, 1998). The genus *Rhammatocerus* is related to other genera such as *Parapellopedon* Jago, 1971 and *Cauratettix* Roberts, 1937.

Since its description, the most important studies on this genus are due to Jago (1971) and Carbonell (1995), but no taxonomical revisions of its entire species have been performed. At present, the genus is composed of 13 species (Assis-Pujol, 1997a 1997b, 1998; Carbonell, 1995), some of them still not clearly defined (Carbonell pers. comm.). The group is characterized by a high intraspecific variation and certain heterogeneity of its external morphology. So, other characters than morphological have been searched to clearly separate the species. As regards the species identification, in tribe Scyllinini, like in other Gomphocerinae, the genitalia allow differentiation between close species in only rare cases (Carbonell, 1995). The phallic complex is an important morphological character among Acrididae but, in Gomphocerinae, it has not a practical value (Carbonell, 1995). The female genitalia, especially the spermatheca, have revealed its utility in identifying some species (Assis-Pujol & Lecoq, 2000). Furthermore, molecular studies have not provided information helping species identification (Loreto et al., 2008).

As pointed before, the sounds produced by Orthoptera are of great taxonomic value. They play a well-known role as identification system during mating, and for this reason they are of great value for establishing the real status of local populations that display small morphological differences (Blondheim, 1990; García et al., 1995). The study of sounds produced by Gomphocerinae has repeatedly demonstrated its utility to solve species identification problems (Ragge & Reynolds, 1998); however the acoustic behaviour of Neotropical Gomphocerinae has been studied to a very limited extent (COPR, 1982; García et al., 2003; Lorier, 1996; Lorier et al., 2010; Riede, 1987).

*Rhammatocerus pictus* (Brunner, 1900) and *Rhammatocerus brunneri* (Giglio-Tos, 1895) are robust species inhabiting wet high pastures in northern and central South America, between about latitudes 15 and 40 South. *Rhammatocerus pictus* is distributed in central and southern Brazil (Rio Grande do Sul), Paraguay, Chile (Malleco), northern and central Argentina, Bolivia and Uruguay (Assis-Pujol, 1998; Carbonell et al., 2006) and *Rhammatocerus brunneri* in central-southern Brazil, Paraguay, Bolivia and Uruguay (Assis-Pujol, 1997a). In Uruguay, both species occupy similar habitats (low humid areas covered with tall and dense grass and low hill slopes). They are distributed in the country (Assis-Pujol, 1998) and are more frequently found north of Río Negro. Both species are sympatric in Artigas, Rivera and Lavalleja Departments (Assis-Pujol, 1997a, 1998).

These two species are systematically close to each other, having been separated by Assis– Pujol (1998) just on the basis of morphological criteria: colour of hind femora and tibiae and the spermatheca shape. Their acoustic behaviour is up to date unknown.

Our objective is to establish the real status of these two taxa at the light of their sound production and acoustic behaviour, describing the songs produced in different behavioural situations, the behavioural units identified and the sound-morphological structures.
