The food preference of Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015, a species that seems to be spreading across Brazil’s human-modified environments, was investigated. Extensive experiments led to the conclusion that the generalized diet of this species may have facilitated its dispersal. The analysis of 132 feeding records of 44 geoplaninid species revealed a tendency for closely related species to feed on individuals from similar taxonomic groups, suggesting that in this group behavioral evolution is more conserved than phylogenetic diversification.

Diet, flatworm, Geoplaninae, predation, soil fauna.

Land planarians, or Geoplanidae (Platyhelminthes: Tricladida), are nocturnal predators that are common in humid forests (Graff 1899, Froehlich 1966, Winsor et al. 1998), although avoiding wet places (Froehlich 1955).

The general knowledge about these organisms is still far from satisfactory, especially with regards to their biology (Winsor et al.1998, 2004, Prasniski and Leal-Zanchet 2009). In part, this might owe to the fact that researchers tend to give more attention to taxonomic topics than to the behavioral aspects of their subjects (Ogren 1995). Additionally, the difficulties involved in rearing flatworms are a complicating factor (Mcdonald and Jones 2013).

Studies on the feeding preferences of land planarians have, for the most part, focused on species that are found out of their natural distribution range. They have received this kind of attention as a result of their potentially detrimental impact on native faunas. This is the case of the bipaliinids Bipalium kewense (Moseley, 1878), B. adventitium Hyman, 1943, the rhynchodeminids Platydemus manokwari Beauchamp, 1962, Arthurdendyus triangulatus (Dendy, 1894), Endeavouria septemlineata (Hyman, 1939) and the geoplaninid Obama nungara Carbayo et al., 2016 (Dindal 1970, Ducey et al. 1999, Sugiura et al. 2006, Sugiura 2009, Blackshaw 1997, Zaborski 2002, Boll and Leal-Zanchet 2015, Carbayo et al. 2016). The food preferences of the Neotropical Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015, have also been studied (Boll and Leal-Zanchet 2015). The distributional area of this species was seemingly modified by human intervention within the Neotropical region, and its likely geographical areas of expansion include a northern population found at Parque Ecológico do Tietê (São Paulo, São Paulo state, Brazil). Formerly a flood plain of the Tietê River, the plain was converted into a man-created forest area in the 1970s. The nearest locality where the species is found is located ca. 500 km to the south.

Additional studies including data on the diet of native or exotic land planarians can be found in the literature. These data might provide insights on the food preferences of congeners, even though their diet has not been addressed yet.

In the present study, (a) further experiments were carried out to study the diet preferences of the northern population of O. anthropophila, and (b) data on their diet was compiled from field and laboratory observations, as well as from the literature, and mapped onto a genus-level classification of land planarians of the subfamily Geoplaninae, in order to evaluate the behavioral evolution of these animals during their phylogenetic diversification.

We focused on land planarians of the subfamily Geoplaninae. Data on their diet were obtained from three sources: (a) sets of trials and experiments under laboratory conditions using O. anthropophila, (b) our own occasional observations in nature and under laboratory conditions, and (c) published literature.

Nineteen specimens of O. anthropophila were collected during the day between May, 2011 and February, 2012, from the Parque Ecológico do Tietê, in the city of São Paulo, state of São Paulo, Brazil (23°29’06”S 46°31’12”W). The 1400 ha. park is a managed area comprised of meadows, patches of flowers and trees, mainly exotic species. At the time of capture, most individuals were photographed with a digital camera and the length of each specimen was measured. Each specimen was then placed in a labeled cylindrical plastic container with a cotton ball moistened with mineral water, and 2-3 pieces of tree bark for refuge. The containers measured either 10 x 5 cm (diameter x height) (those were allocated for specimens less than 2 cm long), or 11 × 7 cm (allocated for specimens larger than 2 cm). The containers were kept in a cardboard box to provide darkness, at room temperature and humidity, in the Laboratório de Ecologia e Evolução. The containers were cleaned twice a week, with the cotton balls substituted with fresh ones. Five different species of invertebrates that are abundant at the sampling site and coexist with O. anthropophila, were offered as potential prey. Some prey were known to be food items of O. anthropophila (land planarians and mollusks) (Froehlich 1956, Boll et al. 2015), whereas other prey had not been previously recorded as such (Diplopoda). Prey that could not be identified at the species level was identified by its common name and at a higher taxonomic level.

In order to stimulate readiness to feed (Sugiura 2009), specimens were starved for seven days (in a few cases four days) between one set of trials and the next. A trial consisted of putting a specimen of O. anthropophila and a potential prey item together in a 9-14 cm diameter glass Petri dish dampened with a few drops of mineral water. The trials were conducted separately, during the day, and at room temperature, with all available flatworms. In each case, a specimen of any one of the following five potential prey species was offered: diplopods (Arthropoda), slugs (Limacidae, Mollusca), and individuals from three geoplaninid land planarian species, Issoca rezendei (Schirch, 1929), Geoplana quagga Marcus, 1951 and Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006). Interest in the prey was observed and recorded for 30 minutes (Sony Handycam HDR – XR550V camcorder). Feeding was assumed to have occurred if the prey was either totally or partially ingested. If there was no interest, the same procedure was repeated with the same flatworm and a second potential prey by another species. If the situation persisted, the procedure was repeated again and again until all five potential prey items had been offered. In case of ingestion, the trial was considered completed, and no further offerings were made on the same day. The flatworm was then returned to the darkened container. In the first trial, the sequence of prey offerings was always as follows: a millipede, the slug Deroceras laeve (Müller, 1774), and then sequentially three land planarians, I. rezendei, G. quagga, and L. ernesti. A set of trials was considered concluded after the fifth offering was completed, or when prey had been ingested. The following week, a new set of trials took place with all flatworms available; first by offering the second prey of the above-established sequence, followed by the third, fourth, fifth and first, in this sequence. One hundred and two trials were performed in this manner. With the exception of one specimen, the number of days each snail survived in the lab was registered (Table 1). Causes of death were not investigated in this study.

Additionally, 25 experiments involving O. anthropohila with ten different options of prey were carried out. In this case, flatworms that had survived previous trials were starved for 10 days before being offered prey, in accordance with the established procedure.

A summary was compiled with the available diet records and prevailing conditions of diet, both from our own field and lab notes taken since the 1950s (Tables 2, 3), as well as from the literature (Suppl. material 1, 2). Our field notes were originated from several encounters with flatworms feeding on prey. Laboratory notes refer to anecdotal feeding experiments conducted with land flatworms prior to killing them for taxonomic studies. Such experiments consisted of putting a flatworm specimen together with an available potential prey item (which had been found nearby in the field) on a Petri dish or in a small box, followed by an observation of its behavior for 1-5 minutes. If the planarian displayed interest in the prey by attacking it and protruding the pharynx, the prey was taken away for further identification and was considered a food item. In most cases, anecdotal experiments were conducted just once with a land planarian species. Temperature, humidity or times of the day were not controlled.

Specimens of O. anthropophila survived up to 150 days under laboratory conditions (median 20 days; range 6–150 days; Table 1). Four out of the five different types of prey in the initial trials were ingested: the three geoplaninids, and the gastropod. The millipede was always rejected. Predation rate varied from 56.7% (I. rezendei, Fig. 1) to 27.3% (Geoplana quagga) (Table 1; Suppl. material 3). In the additional experiments, O. anthropophila also accepted two additional species of Geoplaninae (Geobia subterranea Schultze & Müller, 1857, and Endeavouria septemlinetata (Hyman, 1939), a ribbon worm (Nemertea) and an exotic gastropod (Bradybaena similaris (Ferussac, 1821)) (Table 2). In constrast, it rejected one species of land planarian (Obama burmeisteri (Schultze & Müller, 1857)), and one species each of veronicellid gastropods, millipedes, woodlice, Mycetophilidae larva, and Microlepidoptera and earthworm larvae, respectively (Table 3).

Data were obtained on the diets of 44 species of Geoplaninae. A total of 132 records (45 new, Tables 1, 2; 87 from the literature, Suppl. material 1) account for the consumed prey, and 244 records mention prey that were not eaten (46 new, Tables 1, 3; 198 from literature, Suppl. material 2).

Nine species of Obama predate on gastropods, and six on other land flatworms. However, O. anthropophila preyed upon nine species across three phyla, including three introduced species, Deroceras laeve (Müller, 1774) from North America (Hammond 1996), Bradybaena similaris (Férussac, 1821) from Asia (Carvalho et al. 2008) and E. septemlineata, probably from Hawaii (Hyman 1939) (Tables 1, 2). Interestingly, according to anecdotal observations (see Table 3), O. carrierei, O. carrierei var. brownish, O. ferussaci, O. josefi and O. trigueira only prey on other geoplaninids (Tables 2, Suppl. material 1) as observed by Boll and Leal-Zanchet (2016).

Matuxia tuxaua (E.M. Froehlich, 1955) and M. matuta (E.M. Froehlich, 1955) feed on beetle larvae, and four species of Luteostriata as well as I. rezendei feed exclusively on woodlice, rejecting any other kind of prey (Prasniski and Leal-Zanchet 2009; Tables 2, Suppl. material 2).

Two species of Cephaloflexa feed on harvestmen, while one of them, C. bergi, also takes other arthropods, such as Diptera and woodlice (Tables 2, Suppl. material 1). Three species of Xerapoa consume small arthropods (Suppl. material 1). Finally, Geoplana cf. vaginuloides (Darwin, 1844) and G. cf. chita (E.M. Froehlich, 1956), two of three taxonomically well-established species of Geoplana (G. pulchella is the third), prey on snails (Tables 2, Suppl. material 1).

Table 1Download as CSV 

Survival and predation rate (indicated as preyed/offered) of 19 specimens of Obama anthropophila when fed with one out of five different types of prey species under laboratory conditions. A specimen not given as prey is indicated with a hyphen. See Material and Methods for details.

Specimen identification	Specimen survival (days)	Issoca rezendei (Geoplaninae)	Luteostriata ernesti (Geoplaninae)	Geoplana quagga (Geoplaninae)	Deroceras laeve (Gastropoda)	Millipede (Diplopoda)	# trials
A	150	3/6	1/2	2/6	2/6	0/5	25
C	76	4/6	1/2	1/5	1/2	0/3	18
B	50	2/3	0/1	1/2	2/3	0/2	11
D	50	2/3	2/2	1/3	0/1	0/1	10
X	89	1/2	0/1	–	1/1	0/1	5
J	6	0/1	0/1	0/1	0/1	0/1	5
E	20	0/1	0/1	0/1	0/1	0/1	5
Alpha	70	0/1	2/2	–	–	0/1	4
P	13	0/1	–	0/1	0/1	0/1	4
Z	19	1/1	1/1	–	0/1	–	3
N	?	1/1	–	0/1	–	–	2
B2	20	1/1	0/1	–	–	–	2
B1	19	0/1	–	0/1	–	–	2
O	6	1/1	–	–	–	–	1
L	6	–	–	–	1/1	–	1
I	6	–	1/1	–	–	–	1
H	~20	1/1	–	–	–	–	1
G	20	–	–	1/1	–	–	1
F	~20	–	–	–	1/1	–	1
		56.7% (17/30)	53.3% (8/15)	27.3% (6/22)	42.1% (8/19)	0.0% (0/16)	102

Table 2.Download as CSV 

Feeding preferences of species of Geoplaninae land planarians presented here for the first time. Observation conditions, either in nature (nat) or in the laboratory (lab), and references are given in brackets; a hyphen indicates observation conditions as unavailable.

Land flatworm species	Invertebrate consumed
Cephaloflexa bergi (Graff, 1899)	Harvestman Progonyleptoidellus striatus (Roewer, 1913) (Gonyleptidae, Opiliones) [lab]
	Harvestman Neosadocus sp. (Gonyleptidae, Opiliones) [nat]
	Harvestman Mischonyx cuspidatus (Roewer, 1913) (Gonyleptidae, Opiliones) [lab]
	Unidentified insect larva [nat]
	Unidentified harvestman (Fig. 4) [nat]
Cephaloflexa bergi (Graff, 1899)	Cricket Gryllus sp. (Orthoptera, Insecta) [lab]
	Unidentified woodlice (Oniscidea, Crustacea) [-]
	Larva of Tenebrio sp. (Coleoptera) [lab]
Geobia subterranea Schultze & Müller, 1857	Unidentified earthworm (Oligochaeta, Annelida) [nat] (Fig. 5)
Geoplana beckeri Froehlich, 1959, incertae sedis	Unidentified earthworm (Oligochaeta, Annelida) [-]
Geoplana cf. chita Froehlich, 1956	Unidentified land snail (Gastropoda) [nat] (Fig. 6)
Geoplana goetschi Riester, 1938 incertae sedis	Unidentified land leech (Hirudinea, Annelida) [nat]
Geoplana quagga Marcus, 1951, incertae sedis	Torn land slug Limax sp. (Gastropoda) [lab]
	Land planarian Dolichoplana striata Moseley, 1877 (Geoplanidae) [lab]
	Unidentified woodlice (Oniscidea, Crustacea) [lab]
Imbira marcusi Carbayo et al., 2013	Unidentified earthworm (Oligochaeta) [nat] (Fig. 7)
Issoca rezendei (Schirch, 1929)	Unidentified woodlice (Oniscidea, Isopoda) [lab]
Luteostriata caissara (E. M. Froehlich, 1955)	Unidentified woodlice (Oniscidea, Isopoda) [lab]
Luteostriata ernesti (Leal-Zanchet & E. M. Froehlich, 2006)	Unidentified woodlice (Oniscidea, Isopoda) [lab]
Luteostriata sp.	Unidentified woodlice (Oniscidea, Isopoda) [lab]
Notogynaphallia plumbea (Froehlich, 1959)	Unidentified larva of an insect [nat]
Notogynaphallia sexstriata (Graff, 1899)	Insecta: larva of an insect [nat] (Fig. 8)
Obama anthropophila Amaral, Leal-Zanchet & Carbayo, 2015	Land planarian Issoca rezendei (Schirch, 1929) (Geoplanidae) [lab] (Fig. 1, Suppl. material 1)
	Land planarian Luteostriata ernesti (Leal-Zanchet & Froehlich, 2006) (Geoplanidae) [lab] (Suppl. material 1)
	Land planarian Geoplana quagga Marcus, 1951, (Geoplanidae) [lab]
	Land planarian Geobia subterranea Schultze & Müller, 1857 (Geoplanidae) [lab] (1/2)
	Land planarian Endeavouria septemlineata (Hyman, 1939) (Geoplanidae) [lab] (1/4)
	Unidentified Terrestrial ribbon worm (Nemertea) [lab] (1/1)
	Land slug Bradybaena similaris (Gastropoda) (1/2)
	Land slug Deroceras laeve (Müller, 1774) (Gastropoda) [lab] (Suppl. material 1)
	Land snail Bradybaena similaris (Férussac, 1821) (Gastropoda) [lab]
Obama burmeisteri (Schultze & Müller, 1857)	Land snail Achatina fulica Bowdich, 1822 (Gastropoda) [nat] (Fig. 2)
	Land slug Bradybaena similaris (Férussac, 1821) (Gastropoda) [lab]
Obama baptistae (Oliveira & Leal-Zanchet, 2012)	Land snail (Gastropoda) [lab]
Obama carinata (Riester, 1938)	Land snail (Gastropoda) [lab]
	Land slug Veronicellidae (Gastropoda) [lab, nat] (Fig. 3)
Obama evelinae (Marcus, 1951)	Unidentified land snail [nat] (Fig. 9)
Obama ferussaci (Graff, 1897)	Land planarian Cephaloflexa bergi (Graff, 1899) (Geoplanidae) [nat]
Paraba franciscana (Leal-Zanchet & Carbayo, 2001)	Unidentified land leech (Hirudinea, Annelida) [nat]
Paraba phocaica (Marcus, 1951)	Land snail Happia sp. (Systrophiilidae, Gastropoda) [nat]
Pasipha tapetilla (Marcus, 1951)	Unidentified millipede [nat]

Figures 1–9. 

Photographs of land planarians feeding under laboratory conditions (1) and in nature (2–9). Orange arrowheads point to prey; blue arrowheads point to the predator flatworm. (1) Obama anthropophila capturing Issoca rezendei. Note the body margins bending onto the prey, thus preventing it from escaping even before being eaten (inset); (2) a young specimen of Obama burmeisteri feeding on the giant African snail (Ilhabela, SP); (3) Obama carinata handling a slug (Veronicellidae) (Parque Estadual da Serra da Cantareira, São Paulo, SP); (4) Cephaloflexa bergi eating a harvestman (Reserva Biológica Augusto Ruschi, ES); (5) Geobia subterranea eating an earthworm (Caxambu, MG); (6) Geoplana cf. chita disturbed while eating a snail to show the shell (São Sebastião, SP); (7) two specimens of Imbira marcusi simultaneously attacking an earthworm (Parque Estadual Intervales, SP); (8) Notogynaphallia cf. sexstriata eating an insect larva (Parque Estadual da Serra do Mar, SP); (9) Obama evelinae eating a land snail (Reserva Natural Municipal Nascentes de Paranapiacaba, Santo André, SP).

Table 3.Download as CSV 

Invertebrates not eaten by species of Geoplaninae land planarians in laboratory conditions, presented here for the first time. If available, number of experiments is given in parentheses.

Land flatworm species	Invertebrate not eaten
Geoplana quagga Marcus, 1951, incertae sedis	Unidentified woodlice (Oniscidea, Crustacea)
	Unidentified millipede (Diplopoda)
	Unidentified springtail (Collembola, Hexapoda)
	Unidentified earthworm (Oligochaeta)
	Unidentified ant (Formicidae, Insecta)
	Unidentified land snail (Gastropoda)
Imbira marcusi Carbayo et al., 2013	Unidentified land snail (Gastropoda)
	Unidentified springtail (Collembola, Hexapoda)
Issoca rezendei (Schirch, 1929)	Unidentified earthworm (Oligochaeta)
	Unidentified land snail (Gastropoda)
	Unidentified torn slug (Gastropoda)
	Unidentified torn springtail (Collembola, Hexapoda)
	Unidentified millipede (Diplopoda)
Luteostriata ernesti (Leal-Zanchet & E. M. Froehlich, 2006)	Unidentified ant (Formicidae, Insecta)
	Unidentified land snail (Gastropoda)
	Unidentified torn springtail (Collembola, Hexapoda)
	Unidentified earthworm (Oligochaeta)
	Unidentified millipede (Diplopoda)
	Unidentified land slug (Veronicellidae, Gastropoda)
	Unidentified earthworm (Oligochaeta)
	Unidentified woodlice (Oniscidea, Crustacea)
	Unidentified millipede (Diplopoda)
	Unidentified larva of Mycetophilidae (Insecta)
	Unidentified larva of Microlepydoptera (Insecta)
Obama anthropophila	Land planarian Obama burmeisteri (Schultze & Müller, 1857) (4)
	Unidentified Veronicellidae (Gastropoda) (1)
	Unidentified larva of Microlepidoptera (Insecta) (1)
	Unidentified larva of Mycetophilidae (Insecta) (1)
	Unidentified land isopod (Oniscidea) (3)
	Unidentified earthworm (Oligochaeta) (4)
Obama braunsi (Graff, 1899)	Unidentified earthworm (Oligochaeta)
	Unidentified woodlice (Oniscidea, Crustacea)
Obama burmeisteri (Schultze & Müller, 1857)	Unidentified woodlice (Oniscidea, Crustacea)
	Unidentified ant (Formicidae, Insecta)
Obama nungara Carbayo, Álvarez-Presas, Jones & Riutort, 2016	Unidentified, very juvenile land snail (Helicidae, Gastropoda)
Pasipha pasipha (Marcus, 1951)	Unidentified land snail (Gastropoda)
	Unidentified earthworm (Oligochaeta)
	Unidentified woodlice (Oniscidea, Crustacea)
	Unidentified ant (Formicidae, Insecta)
Xerapoa pseudorhynchodemus (Riester, 1938)	Unidentified earthworm (Oligochaeta)
	Unidentified torn land slug (Gastropoda)
	Unidentified land snail (Gastropoda)
	Unidentified springtail (Collembola, Hexapoda)
	Unidentified millipede (Diplopoda)
	Unidentified ant (Formicidae, Insecta)

Apparently, geoplaninids prey on a wide range of taxonomic groups, including snails, slugs (Mollusca: Gastropoda), acari, harvestmen (Arthropoda: Arachnida), woodlice (Arthropoda: Crustacea), adult and larval stages of insects (Arthropoda: Hexapoda), land planarians (Platyhelminthes: Tricladida), land leeches, earthworms (Annelida: Oligochaeta), and terrestrial ribbon worms (Nemertea). Some land flatworm species only feed on organisms of one taxonomic group (e.g., Gastropoda, Opiliones), whereas others display wider diet breadth. The former are called ‘specialists’, and the latter ‘generalists’, according to the Ali and Agrawal (2012) grouping of insect herbivores.

We are grateful to the Administration of Parque Ecológico do Tietê for allowing invertebrate sampling. We thank José Horacio Grau for making Fig. 4 available. We thank Ana Laura Santos Almeida, Celso Barbieri, Júlio Pedroni, Marcos Santos Silva and Ricardo Miranda for their help collecting. Thanks to Ana Cristina Vasconcelos, Vinícius Gasparino and Geison Castro (EACH, USP) for histological processing, and Marcos Santos Silva (EACH, USP) for data on prey. We are grateful to Luiz Ricardo L. Simone (MZUSP) for mollusk identification, to José Einicker Lamas, Camila Conti, Marcelo Duarte and Nalva (MZUSP) for insect identification, and to Marcos Ryotaro Hara and Guilherme Pagoti (EACH-USP) for harvestmen identification. We thank Hugh Jones for his kind revision of a previous version of the manuscript. We are indebted to the anonymous reviewers for their valuable comments and suggestions to the manuscript, and to Ramon A. Clark, Christopher Quinn and Domingo Lago-Barcia for revision of the English text. AC acknowledges USP for an undergraduate fellowship. FC has financial support from FAPESP (proc. 2014/13661-8).