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ISSCA

International Society for the Study

and Conservation of Amphibians

(International Society of Batrachology)

SEAT

Laboratoire des Reptiles et Amphibiens, Muséum national d'Histoire naturelle,

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BOARD FOR 1995

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Source : MNHN, Paris

AIRNTTES

INTERNATIONAL JOURNAL OF BATRACHOLOGY

June 1995 Volume 13, N° 2

Alytes, 1995, 13 (2): 45-51. 45

Una nueva especie de Telmatobius

(Anura, Leptodactylidae)

de la ceja de montaña de La Paz (Bolivia)

E. O. LAviLLa * & P. ERGUETA S. **

* Instituto de Herpetologia, CONICET, Fundaciôn Miguel Lillo, Miguel Lillo 251,

4000 Tucumän, Argentina

** Colecciôn Boliviana de Fauna, Instituto de Ecologia, Universidad Mayor de San Andrés, La Paz, Bolivia

A new species of Telmatobius is reported from the humid regions of the

“ceja de montaña” of Kkota Pata, Department of La Paz, Bolivia. This species

is characterized by its large eyes and the structure of the skin, with wide and

protruded semicircular warts.

Bibliothèque Centrale Muséi

00111601 0

En la batracofauna de Bolivia se han registrado hasta el momento tres especies de

Telmatobius que son habitantes estrictos de cursos de agua en bosques de montañas:

Telmatobius bolivianus Parker, 1940, T. verrucosus Werner, 1899 y T. yuracare De La Riva,

1994. Por otra parte, T. simonsi Parker, 1940, descripta originalmente para Sucre, en âreas

de valles mesotérmicos, fue citada por DE LA Riva (1990, 1994) en regiones boscosas de

La Siberia, en el Departamento Cochabamba, aunque esta identificaciôn es dudosa.

Entre las formas estrictamente silvicolas, el estado taxonomico de T. verrucosus es

confuso. VELLARD (1951) consideré a T. bolivianus como un sinénimo mäs reciente de esta

especie, y posteriormente (VELLARD, 1970), consider a ambas como subespecies de

Telmatobius marmoratus. En un trabajo reciente (LAvVILLA & DE LA Riva, 1993), basados

en caracteres larvales, se determiné que Telmatobius bolivianus es una buena especie, y la

posiciôn de T. verrucosus es todavia un interrogante abierto.

En un estudio sobre los anfibios de la regiôn de Kkota Pata, en el Departamento La

Paz, encontramos una poblaciôn de Telmatobius en un arroyo de la ceja de montaña, con

Source : MNHN, Paris

46 ALYTES 13 (2)

caracteres notablemente diferentes a los de los restantes Telmatobius conocidos, y en

especial de aquellos que habitan los cuerpos de agua de los bosques de montaña en Bolivia,

lo que nos leva a describirla como nueva.

Finalmente, conviene destacar que las selvas de montaña del oeste de América de Sur

parecen albergar un interesante conjunto de novedades dentro del género Telmatobius, tal

como se desprende de la sintesis presentada por LAURENT (1980) para Argentina y de los

trabajos de DE LA Riva (1990, 1994) para Bolivia y WIENS (1993) para el norte de Perü.

MÉTODOS

La descripciôn que se presenta a continuaciôn ha considerado los caracteres cläsicos

de morfologia externa. El material estudiado fue fijado en formol 10 % en camara hümeda

durante 24 horas y conservado en alcohol etilico de 70°. Un ejemplar macho adulto fue

teñido y diafanizado siguiendo la técnica propuesta por WassERSUG (1976); aunque en el

presente trabajo no se presenta una descripciôn del esqueleto, un caräcter osteolôgico, la

estructura del hümero, ha sido empleado para diferenciar la nueva especie de Telmatobius

Juracare.

Las medidas fueron tomadas bajo lupa binocular, empleando un calibre con precision

de 0,02 mm.

RESULTADOS

Telmatobius jahuira sp. nov.

(fig. 1)

Holotipo. — Coleccién Boliviana de Fauna (CBF) 01675, macho adulto, colectado por

E. LAviLLA y P. ERGUETA el 21 de abril de 1992 en el Rio Chairo, en las proximidades de

la Mina Copacabana (aproximadamente 16°16'S 67°50'W), Kkota Pata, Departamento La

Paz, Bolivia.

Paratipos. — Alotipo: CBF 01676, hembra adulta, mismos datos que el holotipo. Otro

paratipo: CBF 01571, macho adulto, colectado por R. HINOJOsA y S. OTAZU el 21 de abril

1992 en Kkota Pata, Departamento La Paz, Bolivia.

Etimologia del nombre especifico. — Jahuira es un vocablo aymara que significa rio, y

se lo emplea en alusiôn a los häbitos de esta especie. Aqui es empleado como sustan-

tivo.

Diagnosis. — Telmatobius jahuira est caracterizada por el notable desarrollo de sus ojos,

grandes y sobresalientes, y la piel dorsal con verrugas grandes, hemisféricas, separadas

entre si y pigmentadas de negro, lo que hace que la librea aparezca con numerosas

manchas aproximadamente circulares. Las diferencias con los otros Telmatobius que son

habitantes exclusivos de ambientes selväticos estän puntualizadas en la discusion.

Source : MNHN, Paris

LAVILLA & ERGUETA 47

Fig. 1. — (a) Detalle de la cabeza en vista lateral de Telmatobius jahuira. (b) Vista general del holotipo

de Telmatobius jahuira (longitud hocico-cloaca: 55,6 mm).

Descripciôn del holotipo. — Longitud hocico-ano: 55,6 mm. La cabeza es deprimida, con

la regiôn gular plana; es mâs ancha (21,3 mm) que larga (18,1 mm), con indice cefälico de

0,85.

El hocico es redondeado en vistas dorsal y lateral, y su longitud es menor (4,6 mm)

que el diâmetro del ojo (6,5 mm). El canthus rostralis, marcado, es redondeado y la region

loreal es côncava. Los labios estân levemente engrosados, no son glandulares y se

proyectan por sobre la mandibula inferior en todo su perimetro.

Source : MNHN, Paris

48 ALYTES 13 (2)

Los ojos son proporcionalmente grandes (corresponden a aproximadamente el 36 %

de la longitud de la cabeza) y son muy sobresalientes. La comisura posterior de los

pärpados estä mâs lejos de la boca (6,2 mm) que el margen ventral de los orificios nasales

(4,3 mm). La pupila es redonda y la membrana nictitante es transparente, con una banda

pigmentada, en la que alternan secciones grises y amarillas, en el margen libre. El diämetro

del ojo es mayor que la distancia del ojo a la narina (3,7 mm), y la distancia interocular

anterior (tomada entre las comisuras anteriores de los pärpados) equivale al 47 % del

ancho de la cabeza. Los ojos tienen posicin mäs lateral que frontal.

Desde la regiôn posterior del ojo se extiende, hacia aträs y hacia abajo, un pliegue

supratimpänico engrosado y glandular, que termina a la altura de la implantacion del

miembro anterior. Por debajo de este pliegue y por deträs de la boca se ubica una glândula

postcomisural muy marcada, redondeada, elevada, rugosa y sin cornificaciones.

El timpano y el anillo timpänico no estän diferenciados externamente y la region

timpänica es glandular. Los orificios nasales son subcirculares, rebordeados y no estän

elevados. La distancia internasal (3,5 mm) es levemente menor que la distancia naso-ocular

(3,7 mm), y 2,8 veces menor que la distancia interocular anterior. La lengua es redonda,

entera y libre posteriormente.

La region dorsal del cuerpo y la cabeza presentan numerosas gländulas subcirculares,

proporcionalmente grandes, elevadas y sin cornificaciones, que estän separadas entre si

por äreas rugosas. La regiôn dorsal de los miembros anteriores y posteriores es rugosa y

carece de acümulos glandulares elevados. La region ventral es uniformemente lisa, y no

existen cornificaciones nupciales en el pecho.

El orificio cloacal es posterodorsal (ubicado a la altura de la region media de los

muslos) y no existe un pliegue cloacal definido; en su lugar, toda la region pericloacal

aparece estrechamente plisada. El pliegue supracloacal es breve y recto.

Los dedos de las manos poseen falanges terminales redondeadas y levemente

expandidas. El tubérculo metacarpal interno es oval, estrecho, protuberante y 1,4 veces

mäs largo que el externo, suboval y ancho. Entre ambos tubérculos metacarpales existe un

tubérculo grande, que puede ser interpretado como un tercer tubérculo metacarpal, como

un tubérculo palmar hipertrofiado o como un tubérculo supernumerario del dedo I.

Los tubérculos subarticulares son redondeados, sobresalientes y se disponen segün la

formula I (1), II (1), III (2), IV (2). La palma de la mano es lisa, con escasos tubérculos

redondeados y pequeños. La palmeadura estä reducida a un reborde cutäneo en el margen

externo del pollex. La longitud relativa de los dedos es III > IV > I > II.

Los dedos de los pies Ilevan falanges terminales redondeadas y no dilatadas. El

tubérculo metatarsal interno es oblongo y mayor que el externo, que es oval. No existen

tubérculos plantares, y los tubérculos subarticulares, que en general son ovales,

sobresalientes y enteros, se disponen segün la formula I (1), I (1), III (2), IV (3), V (2).

La palmeadura est mejor desarrollada entre los dedos II-III y III-IV, y alcanza su menor

desarrollo entre los dedos I-IT; en todos los casos, alcanza el extremo de los dedos por

medio de rebordes cutäneos. El pliegue tarsal esta medianamente desarrollado en el

margen interno del hallux, y por deträs del tubérculo metatarsal interno se continüa como

una linea clara. En el margen externo del dedo IV existe un pliegue cutäneo que se extiende

Source : MNHN, Paris

LAVILLA & ERGUETA 49

desde el tubérculo subarticular basal hasta el extremo del dedo. Cuando los fémures son

colocados en ängulo recto con respecto al eje axial del cuerpo, los talones se superponen

y cuando la pata es Ilevada hacia adelante, la articulaciôn tibio-tarsal alcanza la mitad del

ojo. La relaciôn entre las diversas regiones del miembro posterior y la longitud total del

cuerpo son: fémur, 53,2 %; tibia-fibula, 54,1 %; tarso, 27,5 %; pie, 58,4 %.

Los caracteres sexuales secundarios se restringen a las callosidades nupciales que se

ubican en el margen interno y la cara dorsal del pollex, y se presentan como escasas

espinas côrneas que dejan amplios espacios no queratinizados entre si.

Coloraciôn en vida: dorso de cabeza, cuerpo y miembros verde oliva; manchas

aproximadamente circulares negras, de limites netos, en el dorso del cuerpo y la cabeza;

regiôn dorsal de los miembros con manchas irregulares; ventralmente grisäceos, con

manchas amarillo-naranja, ms notorias en los miembros anteriores y posteriores y sobre

el pecho.

Coloracion en fijador: dorsalmente gris oscuro con manchas aproximadamente

circulares negras; ventralmente, gris mediano, con manchas beige.

Notas sobre los paratipos. — Se señalän sélo aquellos caracteres morfolôgicos que divergen

del holotipo, y las medidas correspondientes son presentadas mâs abajo.

Alotipo CBF 01676. — El perfil de la region gular es redondeado. Las manchas

negras del dorso del cuerpo y de la cabeza son mâs numerosas, mayores y algunas pueden

presentar contornos irregulares, por coalescencia de manchas proximas. No existen

rebordes cutäneos en el pollex, y los tubérculos palmares son mâs numerosos y estän mejor

definidos que en el holotipo; existe un tubérculo supernumerario en el dedo I (derecha);

no existen callosidades nupciales. En la pata existe un tubérculo plantar entre los

tubérculos metatarsales.

Otro paratipo CBF 01571. — Con excepciôn de los caracteres morfométricos (ver mäâs

abajo), no existen diferencias morfolôgicas de importancia con el holotipo.

Medidas. — Las medidas estän expresadas en milimetros; el primer valor corresponde al

holotipo, el segundo al alotipo y el tercero al otro paratipo. Longitud del cuerpo: 55.6;

58.9; 55.1. Longitud de la cabeza: 18.1; 18.8; 17.1. Ancho de la cabeza: 21.3; 21.2; 19.

Longitud del hocico: 4.6; 4.6; 4.1. Distancia naso-ocular: 3.7; 3.9; 3.8. Distancia internasal:

3.5; 3.9; 3.7. Distancia interocular anterior: 10.0; 9.2; 9.2. Distancia interocular posterio:

17.9; 18.1; 17.3. Tubérculo metacarpal interno: 5.3; 4.5; 5.6. Tubérculo metacarpal externo:

3.8; 3.8; 4.2. Diämetro del ojo: 6.5; 6.8; 6.8. Diämetro del orificio nasal: 0.68; 0.70; 0.56.

Distancia ojo-boca: 6.2; 5.9; 5.7. Distancia nariz-boca: 4.3; 4.6; 3.9. Longitud del fémur:

29.6; 31.6; 28.9. Longitud de la tibia: 30.1; 32.0; 30.1. Longitud del tarso: 15.3; 15.4; 14.8.

Longitud del pie: 32.5; 34.2; 32.2.

Habitat. — La localidad tipo se encuentra en la region de ceja de montaña del

Departamento La Paz, y el clima regional se caracteriza por presentar un régimen hümedo

a perhümedo. Los datos obtenidos de la estacin meteorolôgica de Chururaqui, en el

vecino valle de Zongo, muestran precipitaciones de 3250 mm anuales.

Holotipo y alotipo fueron coleccionados en las primeras horas de la tarde; estaban

inactivos, en el agua y bajo piedras planas, grandes. El esfuerzo de büsqueda indicaria que

se trata de una especie poco abundante en la localidad tipo.

Source : MNHN, Paris

50 ALYTES 13 (2)

DIsCUSIÔN

Las cuatro especies de Telmatobius reportadas para las selvas de montaña de Bolivia

son, como señaläramos en la introducciôn, Telmatobius bolivianus, T. verrucosus, T.

yuracare y la nueva especie que describimos aqui, T. jahuira.

Telmatobius jahuira se diferencia de T. verrucosus (segün las descripciones disponibles

de WERNER, 1899 y VELLARD, 1951) por presentar: (1) el hocico mäs corto que el diämetro

del ojo; (2) el canthus rostralis mâs marcado; (3) las narinas mâs cerca del ojo que del

hocico; (4) el primer dedo de la mano mäs largo que el segundo; (5) la membrana

interdigital poco desarrollada entre los dedos II-III y III-IV de la pata; (6) el pliegue tarsal

vestigial; (7) la mandibula superior proyectada sobre la inferior en todo su perimetro; (8)

un patrôn de coloracién dorsal con color de base verde oliva y manchas circulares negras.

Telmatobius jahuira difiere de T. bolivianus (segün la descripciones de PARKER, 1940

y VELLARD, 1951) por presentar: (1) el hocico redondeado y mäs corto que el diâmetro

ocular; (2) los dedos de las manos con falanges terminales redondeadas, levemente

expandidas; (3) el primer dedo de la mano mäs largo que el segundo; (4) los dedos II y

III de las manos sin reborde cutäneos; (5) los extremos de los dedos de las patas no

dilatados; (6) la membrana interdigital poco desarrollada entre los dedos II-IIT y III-I

(?) el pliegue tarsal vestigial; (8) la articulaciôn tibio-tarsal alcanzando a la mitad del ojo;

(9) la piel con gländulas muy desarrolladas; (10) un patrôn de coloraciôn diferente; (11)

la estructura, abundancia y disposicién de las espinas corneas en los pulgares de los

machos.

Telmatobius jahuira difiere de T. yuracare (segün la descripciôn de DE LA Riva, 1994),

entre otros caracteres, por: (1) carecer de espina humeral en los machos; (2) la orientacion

de los ojos; (3) la estructura de la piel del dorso del cuerpo; (4) la textura de la piel del

dorso del cuerpo; (5) el patrôn de coloraciôn.

Cualquiera sea la posiciôn taxonémica definitiva de los taxa yungueños del género

Telmatobius en Bolivia, se observa claramente que T. jahuira es una especie diferente de

las previamente descriptas.

Desafortunadamente no contamos aün con larvas de esta especie, por lo que no

podemos atribuirla a ninguno de los dos grupos conocidos (LAviLLA, 1985).

LITERATURA CITADA

DE LA Riva, I. 1990. — Lista preliminar comentada de los anfibios de Bolivia, con datos sobre su

distribuciôn. Boll. Mus. reg. Sci. nat. Torino, 8: 261-319.

= 1994. — À new aquatic frog of the genus Te/matobius (Anura: Leptodactylidae) from Bolivian

cloud forests. Herpetologica, 50 (1): 38-45.

LAURENT, R. F., 1980. — Herpetofauna of the forest remnants of North-Western Argentina. National

Geographic Research Reports, 1977 Projects: 417-427.

Source : MNHN, Paris

LAVILLA & ERGUETA 51

Lavizca, E. O., 1985. — Diagnosis genérica y agrupaciôn de las especies de Telmatobius (Anura:

Leptodactylidae) en base a caracteres larvales. Physis, (B), 43 (105): 63-67.

LaviLLa, E. O. & DE La Riva, L, 1993. - La larva de Telmatobius bolivianus (Anura,

Leptodactylidae). Alytes, 11 (2): 37-46.

PARKER, H. W., 1940. — Percy Sladen Trust Expedition to Lake Titicaca under the leadership of Mr.

H. Cary Gilson, M. A. XII. Amphibia. Trans. linn. Soc. London, 3 (1): 203-216.

VELLARD, J., 1951. — Estudios sobre batracios andinos. I. El grupo Telmatobius y formas afines.

Mem. Mus. Hist. nat. “Javier Prado”, 1: 1-89, 8 läm.

WASsERSUG, R. J., 1976. — A procedure for differential staining of cartilage and bone in whole

formalin-fixed vertebrates. Stain. Tech., 51: 131-134.

WERNER, F., 1899. — Beschreibung neuer Reptilien und Batrachier. Zool. Anz., 22 (602): 479-484.

WIENS, J. J., 1993. — Systematics of the leptodactylid frog genus Telmatobius in the Andes of

Northern Peru. Occas. Pap. Mus. nat. Hist. Univ. Kansas, 162: 1-176.

Corresponding editor: Jaime E. PÉFAUR.

Source : MNHN, Paris

Alytes, 1995, 13 (2): 52-66.

Reassessment of central

Peruvian Telmatobiinae

(genera Batrachophrynus and Telmatobius).

IL. Allozymes and phylogenetic relationships

Ulrich SNscH & Norbert JURASKE

Institut für Biologie, Universität Koblenz-Landau, Rheinau 1, 56075 Koblenz, Germany

Three hypotheses on the phylogenetic relationships among central Peru-

vian Telmatobiinae were tested: (1) the common ancestor of the two

Batrachophrynus species diverged from the telmatobiine stock independentiy

from the common ancestor of the present Telmatobius; (2) B. macrostomus

and B. brachydactylus separated independently from the lineage leading to the

present Telmatobius; (3) the separation of the common Batrachophrynus

ancestor from Telmatobius occurred after the differentiation of the Telmato-

bius stock into geographical lineages. Allozymes clearly indicate the mono-

phvly of Batrachophrynus, and that the southern Peruvian T. culeus is more

closely related to the central Peruvian T. jelskii and T. rimac than to either of

the Batrachophrynus species. AIl available evidence supports the first hy-

pothesis.

INTRODUCTION

Three genera of central Peruvian Telmatobiinae are currently recognized: Telmatobius

Wiegmann, 1835, Batrachophrynus Peters, 1873 and Lynchophrys Laurent, 1983 (FROST,

1985; DUELLMAN, 1993). À taxonomic reassessment of the six species assigned to these

genera indicated that the monotypic genus Lynchophrys is not valid and that Lynchophrys

brachydactyla should be referred to as Batrachophrynus brachydactylus (SiNscH et al.,

1995). In contrast, osteological evidence (LyNCH, 1978) and morphometric data (SINSCH et

al., 1995) continue to support the validity of the genera Batrachophrynus and Telmatobius.

The corresponding phylogenetic hypothesis assumes a monophyletic origin of both genera

(Cet, 1986).

LAURENT (1983), however, proposed an alternative hypothesis. He assumed an early

separation of B. macrostomus, and an independent, but later, separation of B. brachy-

dactylus from the Telmatobius stock. If this was true, the morphometric and osteological

similarities between the two Batrachophrynus species would be convergences and the genus

would be paraphyletic. There is a third possibility, considering the conspicuous general

similarity between the two genera: the separation of the Batrachophrynus ancestor from the

Source : MNHN, Paris

SINSCH & JURASKE 53

central Peruvian Telmatobius stock may have occurred after the splitting of Telmatobius

into a northern and a southern lineage. In this case, the genus Batrachophrynus would be

invalid and its two species would have to be included in the genus Telmatobius.

The aim of our study was to test these alternative hypotheses by analyzing the genetic

similarity among four central Peruvian and a southern Peruvian species of Telmatobiinae.

The stream-dwelling T. jelskii (Peters, 1873), T. rimac Schmidt, 1954 and B. brachydactylus

Peters, 1873, and the lake-dwelling B. macrostomus Peters, 1873, inhabit neighbouring

regions within the Departments of Cerro de Pasco, Junin and Lima. The Titicaca frog T.

culeus (Garman, 1875), which represents another evolutionary lineage within the genus

Telmatobius (VELLARD, 1951, 1953, 1955), was included as a potential outgroup.

Horizontal starch gel electrophoresis of the proteins of blood, muscle and liver

homogenates was used to assess allelic variation in homologous loci among all populations

and species. Our reconstruction of phylogenetic relationships within this Andean group of

Telmatobiinae clearly supports CErs (1986) hypothesis of a monophyletic origin of

Batrachophrynus and Telmatobius.

MATERIAL AND METHODS

We examined 111 frogs representing five species of Telmatobiinae (Leptodactylidae):

(1) Batrachophrynus brachydactylus: 11 males and 11 females from a brook near Ondores

(Dep. Junin, Perü); (2) B. macrostomus: 6 males, 7 females and 8 juveniles from Junin Lake

(Dep. Junin); (3) Telmatobius jelskii: (a) 5 males, 2 females and 14 juveniles from Rio

Shullcas, Palian near Huancayo (Dep. Junin), (b) 6 males and 3 females from Cuyrohuasi

near Tarma (Dep. Junin); (4) T. rimac: (a) 7 males, 7 females and 8 juveniles from Rio

Chillén, Obrojillo near Canta (Dep. Lima, Perü), (b) 9 males, 4 females and 2 juveniles

from Quebrada Huaytara, Canta (Dep. Lima); (5) T. culeus: 1 male from Titicaca Lake

(Dep. Puno, Perü).

Following the morphological classification of specimens (SINsCH, 1986, 1990; SiNscH

et al., 1995), we collected blood samples (about 60 ul per individual) from the vena

angularis of the living frogs (NÔLLER, 1959). Samples were centrifuged at 11,500 rpm for

3.33 min and the cell fraction was dissolved in 20 ul homogenate buffer (tris-EDTA-

NADP at pH 7.0) and stored at -18°C until use. The frogs were sacrificed and liver and

muscle samples were taken and mechanically homogenated in 50-150 pl in tris-EDTA-

NADP buffer at pH 7.0. AIl samples were collected in Perü in 1992 except for the blood

sample of the T. culeus specimen. This individual was kept in the Museum of Natural

History in Bonn, Germany, and was included to look for fixed alleles in the Telmatobius

and Batrachophrynus lineages.

Gels for horizontal electrophoresis were prepared at 12 % with SIGMA starch. Four

buffer systems were used at constant 55 mA and 4°C: (1) tris-citrate (electrode: pH 8.0; gel:

PH 8.7), duration of electrophoresis: 5 h; (2) tris-malate (electrode and gel: pH 8.4), 4h;

(3) tris (electrode and gel: pH 8.6), 5 h; (4) tris-borate (electrode and gel: pH 9), 4 h. Each

Source : MNHN, Paris

s4 ALYTES 13 (2)

gel was sliced into five 2 mm thick slabs for staining. Procedures for staining were those

described by PASTEUR et al. (1988), SHAW & PRASAD (1970) and SHERIF (1990).

Allozymes examined were representative of 12 enzyme systems controlled by a total

of 22 presumptive loci: aspartateamino transferase (1 locus, abbreviation: AAT, E.C. No.

2.6.1.1); adenylate kinase (1, AK, 2.7.4.3); esterase (4, EST, 3.1.1.1); glucosephosphate

isomerase (2, GPI, 5.3.1.9); hexanol dehydrogenase (2, HDH, 1.1.1.56); isocitrate

dehydrogenase (2, IDH, 1.1.1.42); lactate dehydrogenase (2, LDH, 1.1.1.27); malate

dehydrogenase (2, MDH, 1.1.1.37); malic enzyme (1, ME, 1.1.1.40), peptidase (3, PEP,

3.4.1.1); 6-phosphogluconate dehydrogenase (1, 6-PGD, 1.1.1.44); phosphoglucomutase

€, PGM, 2.7.5.1). In addition, we scored the non-enzymatic hemoglobin (Hb).

Multiple loci were numbered from cathode to anode. Presumptive alleles were

designated numerically according to their mobility relative to the most common

electromorph (assigned 100) of the T. jelskii population from Palian. Faster moving

electromorphs were assigned higher values (above 100), slower moving ones lower values

(below 100). For reference, each electrophoretic run included samples of the T. jelskii

population from Palian. Statistical analyses included the calculation of allele frequencies,

of Neïs genetic distance (NEI, 1972), Cavalli-Sforza’s chord distance (CAVALLI-SFORZA &

EbwaRDs, 1967) and Reynolds’s genetic distance (REYNOLDSs et al., 1983) by the program

GENDIST 3.4 (package PHYLIP; FELSENSTEIN, 1985). Average heterozygosity per locus

(Ho = observed frequency; He = expected frequency), proportion of polymorphic loci

(P %), and the mean number of alleles per locus (A) were calculated for each sample

except T. culeus. We used the G-test to detect deviations of observed heterozygosity from

the Hardy-Weinberg equilibrium.

Reconstruction of phylogenetic relationships between the examined populations is

based on four algorithms applied to allele frequencies: (1) UPGMA method (program

NEIGHBOR 3.41); (2) Fitch-Margoliash method assuming equal rates of evolutionary

change in all lineages (KITSCH 3.41); (3) Fitch-Margoliash method without evolutionary

clock (FITCH 3.41); (4) maximum likelihood method (CONTML 4.42). All calculations

are based on the cited programs in the package PHYLIP (FELSENSTEIN, 1985).

RESULTS

ALLELIC VARIATIONS OF PROTEINS

A total of 23 presumptive loci (enzymes: AAT, AK, EST, GPI, HDH, IDH, LDH,

MDH, ME, PEP, 6PGD, PGM; non-enzymatic protein: Hb) was scored in 6 populations

of four central Peruvian telmatobiine species and in one specimen of T. culeus (Table I).

Five loci were monomorphic in all samples: HDH1, LDH2, PEP2.1, 6PGD and PGMI1.

Allele frequencies are listed in Table I. The observed heterozygosity significantly deviated

from the expected heterozygosity of the Hardy-Weinberg equilibrium in all populations

because of a deficit of heterozygotes at all loci (G-test, P < 0.001). The following account

of the loci demonstrates that fixed alleles at the LDH1 locus permit an unequivocal

Source : MNHN, Paris

SINSCH & JURASKE 55

distinction between the genera Batrachophrynus and Telmatobius, and that the southern

Peruvian T. culeus differs from all central Peruvian populations by the presence of an

unusual allele at the MDHI locus.

Aspartateamino transaminase

Following electrophoresis in the tris-malate system, we regularly detected activity in

muscle and liver samples. The Batrachophrynus species are monomorphic for allele 100,

whereas the Telmatobius populations possess a second, more slowly migrating AAT (allele

60). Heterozygotes were not observed.

Adenylate kinase

All blood and liver samples show stainable activity following electrophoresis in the

tris-borate system. The frequencies of the main alleles 100 and 120 are similar in all

samples. One specimen of B. brachydactylus possessed a third allele 75. Heterozygotes were

not observed.

Esterase

We detected four loci which can easily be distinguished by their specific activity in

different tissues and their electrophoretic mobility in the tris system. The most slowly

moving esterase (EST1) usually shows low activity in the liver samples and sometimes also

in the blood samples, whereas EST2 is active in all samples but stains most strongly in the

liver samples. The faster moving esterases (EST3, EST4) produce stainable bands with

about the same activity in all samples. The frequencies of the main alleles 90 and 100 of

the EST1 locus are similar in all populations. Two B. brachydactylus showed a third allele.

Two alleles 90 and 100 of EST2 locus are present in both Batrachophrynus species and in

T. rimac, whereas in T. jelskii this locus is monomorphic. With the exception of two

heterozygotes of the constitution 50/100 (B. brachydactylus) at the EST1 locus, all other

specimens are homozygotes for the four esterase loci.

Glucosephosphate isomerase

In the tris-borate system an anodally migrating GPI was active in blood and liver

samples. Allele 100 dominates in all populations, except for those of T. rimac which are

almost monomorphic for allele 73. No heterozygotes were detected.

Hexanol dehydrogenase

The tris-malate system resolved two systems of HDH, the monomorphic HDH1 locus

being active at similar levels in all tissues and the polymorphic HDH2 locus with stainable

Source : MNHN, Paris

56 ALYTES 13 (2)

activity in muscle and liver samples. The frequency of allele 100 is greater in the

Batrachophrynus species than in the Te/matobius species where allele 75 dominates. A

single heterozygote of the constitution 75/100 (B. macrostomus) was detected.

Isocitrate dehydrogenase

Two polymorphic loci coding for enzymes of considerably different electrophoretic

mobility were detected in the tris-citrate system. Both loci were almost exclusively active

in muscle and liver samples. In Telmatobius the IDH1 locus is monomorphic for the allele

100, whereas in Batrachophrynus a second, rare allele 93 is present. Four alleles are found

at the IDH2 locus with allele 100 dominating in Te/matobius. In Batrachophrynus allele 90

is the most common one. A single heterozygote of the constitution 100/125 (8.

brachydactylus) was detected.

Lactate dehydrogenase

Following electrophoresis in the tris-citrate system, only one band stained in the

muscle and liver samples of all species, whereas in the blood samples up to five bands

appeared. The common stainable band of all tissues is the tetramer of the unit coded for

by the LDHI locus. The five banded pattern was detected only in the blood of the two

Batrachophrynus species, whereas in the blood of the Te/matobius species at most three

bands stained, corresponding to slowly moving tetramers. The LDHI1 locus is diagnostic

for the distinction of the two genera: all Telmatobius are fixed for the allele 100, all

Batrachophrynus for the allele 33. Pure tetramers of the unit coded for by the LDH2 locus

were found in both Batrachophrynus with the same electrophoretic mobility (allele 100).

The poor resolution of the one or two bands of mixed tetramers in Telmatobius does not

permit a reliable estimate of the position of the non-expressed pure LDH2-tetramer.

Malate dehydrogenase

The best resolution of the bands corresponding to two polymorphic MDH loci was

found in the tris-borate system. The MDHI1 locus produces a slowly moving protein which

was exclusively active in blood and liver samples. In contrast, the faster moving product

of the MDH2 locus was present in all tissues, but, especially in the muscle samples,

subbands frequently appeared which were not present in the other tissues of the same

individual. The MDHI locus is diagnostic for the southern Andean T. culeus which

possesses allele 125, whereas only alleles 71 and 100 are present in all central Peruvian

populations. The frequencies of allele 71 are extremely low in both Batrachophrynus, but

considerably greater in Telmatobius. Three alleles are found at the MDH2 locus, the

usually dominating allele 100, the less frequent allele 85 and the rare allele 115. Only one

heterozygote of the constitution 85/100 (T. rimac, Obrojillo) was detected at the MDH2

locus.

Source : MNHN, Paris

SINSCH & JURASKE 5:

Malic enzyme

The polymorphic ME locus accounted for regular activity in all samples following

electrophoresis in the tris-malate system. Allele 100 dominates in all populations and

reaches almost monomorphic frequencies in T. jelskü. T. rimac and the two Batracho-

phrynus differ from this species by a considerably higher frequency of allele 60 and the

presence of the rare allele 20. At this locus we detected 12 heterozygotes: six of the

constitution 20/60 (3 B. brachydactylus, 1 B. macrostomus, 2 T. rimac from Obrojillo), and

another six of the constitution 60/100 (1 B. brachydactylus, 1 B. macrostomus, 3 T. rimac

from Obrojillo, 1 T. rimac from Huaytara).

Peptidase

Following electrophoresis in the tris-citrate system, we identified three PEP loci which

were monomorphic in most populations. The peptidase of the PEP1 locus specifically

digested the dipeptide VAL-LEU and had greater activity in blood and liver samples than

in muscle tissue. The other two peptidases used the tripeptide LEU-GLY-GLY as a

substrate, but the activity of the PEP2.1 locus was restricted to muscle and liver samples

of B. brachydactylus and T. rimac and one specimen of T. jelskii, whereas the PEP2.2 locus

was exclusively active in the blood samples of all populations (except T. culeus). The PEP1

locus is monomorphic in all but one species: B. macrostomus possesses a second allele 83

in low frequency. The PEP2.2 locus is monomorphic in all Telmatobius populations, but

in the two Batrachophrynus species a second allele 115 occurs in low frequency. No

heterozygotes were found at any of the loci.

6-phosphogluconate dehydrogenase

Following electrophoresis in the tris-citrate system, in all tissues we found activity

corresponding to the same allele of a monomorphic locus.

Phosphoglucomutase

Enzyme systems corresponding to two loci were resolved in the tris-malate system.

The monomorphic PGMI locus stained with similar activity in all tissues, whereas the

polymorphic PGM2 locus was detectable exclusively in the muscle and liver samples. In

Telmatobius and B. brachydactylus allele 80 dominates, in B. macrostomus allele 100. A

total of 6 heterozygotes was found: three of the constitution 80/100 (T. rimac from

Obrojillo), one of 80/113 and two of 100/113 (T. jelskii from Palian).

Hemoglobin

In the tris system the distinction of bands corresponding to hemoglobin was best. We

found two alleles present in all central Peruvian populations, allele 100 dominating in the

Telmatobius, allele 120 in the Batrachophrynus. Heterozygotes were not detected.

Source : MNHN, Paris

58 ALYTES 13 (2)

Table L. - Allele frequencies at the polymorphic loci in 7 samples of 5 species of Andean

Telmatobiinae (genera Batrachophrynus and Telmatobius). P %: relative frequency of

polymorphic loci; A: average number of alleles per locus; HL: relative frequency of expected

heterozygosity; H: relative frequency of observed heterozygosity.

(Palian) | (Huaytara) | (Obrojillo)

N=15 à 4

Allles | N=22 | N=21 | N=9 | N=21 N=1

AAT 60] - = 0.143 0.059 | 0.938 0.952 | no activity

100] 1.000 1.000 | 0.857 0.941 0.062 0.048 | detectable

AK 75| 0.052 = = -. . 7 A

100| 0.789 0.900 0.571 0.824 0.813 0833 |roaciiy

120] 0.159 0.100 0.429 0.176 0.187 0167» | 401e0table

ESTI 50| 0.077 = - = . = pe

90! 0.154 0.231 = 0.286 0.083 0.125 =.

100! 0.769 0.769 1.000 0.714 0.917 0.875 1.000

EST2 90| 0.105 0.053 = = 0.125 0.111 .

100! 0.895 0.947 1.000 1.000 0.875 0.889 1.000

EST3 94! O.111 0.421 0.429 0.063 0.125 0.167 .

100! 0.889 0.579 0.571 0.937 0.875 0.833 1.000

EST4 92 - 0.053 = 0.067 0.125 0.412 =

100 1.000 0.947 1.000 | 0.933 0.875 0.589 1.000

GPI 73] 0091 0.050 | 0.333 0.278 1.000 0.950 =

100] 0.909 0.950 | 0.667 0.722 5 0.050 1.000

HDH 75] 0.545 0.395 0.750 0.850 0.875 0.579 no activity

100! 0.455 0.605 0.250 0.150 0.125 0.421 detectable

IDH 93) 0.091 0.050 = = = = no activity

100! 0.909 0.950 1.000 1.000 1.000 1.000 detectable

IDH 90! 0.600 0.400 = = = 0.071 L

100| 0.300 = 1.000 0.667 1.000 0.929 | no activity

11 - 0.400 . 2 - - detectable

125] 0.100 0.200 - 0.333 - -

LDHI 33| 1.000 1.000 . . . =. =.

100 - - 1.000 1.000 1.000 1.000 1.000

MDHI 71] 0.050 0.067 0.444 0.188 0.688 0.556 -

100| 0.950 0.933 0.556 0.912 0.312 0.444 =

125 - - - - - - 1.000

MDH2 85| 0111 0.167 0.500 0.400 0.563 0.262 =

100| 0.833 0.750 = 0.600 0.437 0.643 1.000

115, 0.056 0.083 0.500 - - 0.095 "

ME 20| 0.167 0.026 - - ! 0.053 -

60| 0.286 0.474 “ 0.063 0.438 0.395 -

100| 0.547 0.500 1.000 0.937 0.562 0.552 1.000

PEPI 83 = 0.125 =

100] 1.000 | 0.875 1.000 1.000 1.000 1.000 1.000

PEP2.2 100] 0.714 | 0750 | 1000 | 1.000 | 1.000 | 1.000 [noactiviy

115] 0.286 0:250 - - - - detectable

PGM2 80] 0611 0.400 0.857 0.500 1.000 0.750 ee

0.389 0.600 - 0.333 - 0.250 |n0 activity

18] - - 0.143 0.167 £ s detectable

Hb 100] 0.136 1.000

120| 0.864 “

P% 0.61

A 1.80

He 0:30

He 0.04

Source : MNHN, Paris

SINSCH & JURASKE

Table II. - Matrix of genetic distances among six samples of four central Peruvian species of Andean

Telmatobiinae (genera Batrachophrynus and Telmatobius).

A. Neï's genetic distance

B. Cavalli-Sforza's chord distance

Species + B. T. jelskir T. jelskii T: rimac

Species + 3. T. jelskaii T. jelshai T: rimac

Population | macrostomus (Cuyro.) (Palian) (Obrojillo)

Dachpdacphs| 20801 02237 0.1204 0.3177 02613

e E 02304 0.1417 0.3464 0.2874

macrostomus

ne E 0.0661 0.1055 0.1296

(Cuyro.)

ARE - 0.1387 0.1232

(Palian)

T: rimac : NS

(Huaytara)

T. rimac

(Huaytara)

from the data published in WIENS (1993).

Population | macrostomus (Cuyro.) (Palian) (Huaytara) (Obrojillo)

Brachdacyus| 22211 0.5993 0.3667 0.6996 0.5434

B: ; 0.6621 0.4227 0.7768 0.6131

macrostomus

T. jelskii - 0.2349 0.3092 0.3408

(Cuyro.)

T. jelski - 0.3123 0.2641

(Palian) l :

T. rimac p 0.803

(Huaytara) :

C. Reynolds’s genetic distance

Species + B. T. jelskii T. rimac

Population | macrostomus (Palian) (Obrojillo)

Brachydactytus | 0895 0.4575 0.3009 0.5494

B. n 0.4443 0.3174 0.5458

macrostomus

T.jelskii

- 0.2238 03512

(Cuyro.)

T. jelskii ; 03840

(Palian) À

T. rimac

Table III. - Matrix of Neï's genetic distances among four north Peruvian Te/matobius species, calculated

Species Telmatobius latirostris | Telmatobius necopinus | Telmatobius truebae

Telmatobius brevipes 0.9246 0.8504 0.8702

Telmatobius latirostris - 0.4479 0.3411

Telmatobius necopinus - 0.4973

Source : MNHN, Paris

60 ALYTES 13 (2)

Batrachophrynus Telmatobius

brachy. macro. jelskii rimac

_—. € P H 0

(=)

où

[e}

Neïs genetic distance

0.15

Fig. 1. — UPGMA dendrogram of genetic similarity among six samples of four central Peruvian

species of Andean Telmatobiinae, based on the matrix of Neï’s genetic distances (Table II A).

Batrachophrynus Telmatobius

brachy. macro. jelskii rimac

600) P H 0

0.05

0.10

0.15

0.20

Cavalli Sforza's chord distance

0.25

0.30

Fig. 2. — UPGMA dendrogram of genetic similarity among six samples of four central Peruvian

species of Andean Telmatobiinae, based on the matrix of Cavalli-Sforza’s chord distances (Table

II B).

Source : MNHN, Paris

SINSCH & JURASKE 61

Batrachophrynus Telmatobius

brachy. macro. jelskii rimac

Ci PM. 9

0.00

e e e

2 2 5

ao o a

Reynold's genetic distance

ES)

0.25

Fig. 3. —- UPGMA dendrogram of genetic similarity among six samples of four central Peruvian

species of Andean Telmatobiinae, based on the matrix of Reynolds’s genetic distances (Table II

©).

Telmatobius rimac

Huaytara

Obrojito 87%

Batrachophrynus Telmatobius

macrostomus jelskii

Pelian

Batrachophrynus

brachydactylus

Fig. 4. — Genetic relationships among six samples of four central Peruvian species of Andean

Telmatobiinae. Due to the absence of an outgroup the tree is arbitrarily rooted at the mean

distance between the most similar populations of Batrachophrynus and Telmatobius. Maximum

likelihood method, based on the allele frequencies (Table I); best tree out of 202 examined in five

runs: In likelihood = 88.3.

Source : MNHN, Paris

62 ALYTES 13 (2)

PHYLOGENETIC RELATIONSHIPS

Allele frequencies (Table I) obtained for the six populations of four central Peruvian

telmatobiine species were used for the calculation of three measures of genetic distances

(Table IT). The reconstruction of the phylogenetic relationships by four commonly used

algorithms was based on either allele frequencies or distance matrices (figs. 1-4).

Independent of the algorithm used for the calculation of the genetic distances among

the populations sampled, the matrices obtained shared the following features: (1) the

genetic differentiation between the two populations of T. rimac and between the two

Batrachophrynus species was low and at the same level; (2) the level of differentiation

between the two T. jelskii populations was 2-3 times greater than between the T. rimac

populations, and the Batrachophrynus species; (3) the Telmatobius species were genetically

more similar to each other than any to the Batrachophrynus species.

All algorithms based on distance matrices produced identical groupings of popula-

tions. Therefore, only the UPGMA-dendrograms are shown as representative examples

(figs. 1-3). The main clusters corresponded to the genera Batrachophrynus and Telmatobius

and conspecific populations were placed together. The unrooted maximum likelihood tree

based on allele frequencies shows a similar grouping of the populations (fig. 4). However,

the T. jelskii populations are placed on different branches of the Te/matobius lineage.

In addition to the four central Peruvian species, we also included the southern

Peruvian T. culeus into the analysis. As the data on the allozymes expressed in this species

are based on one individual, a quantitative approach to the phylogenetic relationships is

not possible yet. Nevertheless, a qualitative reconstruction based on the presence or

absence of alleles was attempted (fig. 5). Four monomorphic loci and several common

alleles at the polymorphic loci justify the common root of the dendrogram. The distinction

between Batrachophrynus and Telmatobius is based on fixed alleles at the LDHI1 locus. T.

culeus differs from T. jelskii and T. rimac by fixed alleles at the MDHI locus. In B.

brachydactylus and B. macrostomus, and in T. jelskii and T. rimac, respectively, we were

unable to find a locus with different fixed alleles. Species distinction is based on the

presence and absence, respectively, of rare alleles at the EST2-locus and the PEP1-locus.

DISCUSSION

The radiation of a late tertiary stock of telmatobiine frogs into the recently uplifted

cordilleran environments led to a differentiation of about 30 presently known species (CE,

1986; DUELLMAN, 1993; FRosT, 1985; WiIENS, 1993). Allopatric populations which inhabit

numerous interandean valleys and streams of the Pacific or Atlantic hydrographic systems

give an idea of the mechanisms of speciation at work. The geomorphologically

complicated Late Pleistocene landscape favoured the formation of disjunct populations

(Ce, 1986). Progressive genetic changes in isolated gene pools probably promoted

allopatric speciation in the Andean Telmatobiinae. The tendency for homozygosity at

almost all loci in the Andean populations already studied is a strong indication of small

Source : MNHN, Paris

SINSCH & JURASKE 63

population size and interrupted gene flow between populations. Large genetic distances

between populations assigned to T. jelskii and inhabiting different river systems

demonstrate that allopatric speciation is still the norm at the present time.

Early attempts to analyze relationships between the Andean Telmatobiinae (VELLARD,

1951, 1953, 1955) mainly reflect groupings assigned by biogeographical convenience to a

poor data base. LyNCH (1978) was the first to use a cladistic approach on a set of

morphological and osteological data. He placed the Andean genera Batrachophrynus and

Telmatobius, along with additional seven genera (A/sodes, Atelognathus, Eupsophus,

Hylorina, Insuetophrynus, Limnomedusa and Somuncuria), into the tribe Telmatobiini

Fitzinger, 1843. All cladograms (based on differing numbers of OTUs) emphasized that

the common ancestor of the lineage leading to the two Batrachophrynus species separated

at a very early stage from the lineage leading to the present Telmatobius species.

Nevertheless, D'UELLMAN’S (1979: 424) statement: “the systematic relationships of the

species of Telmatobius presently are too poorly known to assess fully the historical

biogeography of the group” remains valid in spite of increased knowledge on the Andean

Telmatobiinae accumulated since VELLARD’s pioneer work.

Our attempt to reconstruct phylogenetic relationships of the central Peruvian

Telmatobiinae is based on allozymes. The results correspond to those on northern

Peruvian Telmatobius (21 individuals, 4 species, 19 loci) in terms of low heterozygosity at

most loci (WIEns, 1993). However, the Neï’s distances among the species which we studied

are generally lower than those among four northern Peruvian Telmatobius species (Table

II, calculated from Table 5 in Wiens, 1993). This discrepancy is probably due to

differences in method (e. g., number and kind of loci scored, number of individuals, buffer

systems, resolution of gels). All dendrograms and trees derived from processing allele

frequencies and distance matrices indicate the same sequence of speciation events among

the four species of Batrachophrynus and Telmatobius we examined.

Allozymes clearly support monophyly in Batrachophrynus. The gene pools of B.

brachydactylus and B. macrostomus were surprisingly similar and the genetic distance of

0.03 between the two species is the lowest interspecific Neï’s distance ever reported for

Amphibia (usually 0.1 to 3.0; AvisE & AQUADRO, 1982). LAURENT’s (1983) hypothesis of

an independent derivation from the Telmatobius stock is not supported. This hypothesis

apparently resulted from a misinterpretation of convergent morphological traits in the

stream-inhabiting B. brachydactylus and Telmatobius species.

We were unable to detect any fixed genetic difference between the two Batrachophry-

nus species and even allele frequencies are very similar. Taken alone, allozymes would

suggest that B. brachydactylus and B. macrostomus are conspecific. On the other hand,

these taxa are morphologically well defined, extremely different in size, and they live in

different habitats (PETERS, 1873; SINSCH, 1990; SinscH et al., 1995). Intermediate

individuals between the brachydactylus phenotype and the macrostomus phenotype are not

known. Personal field observations in the area of Lake Junin, where both taxa occur

sympatrically, did not yield any evidence of interbreeding. In conclusion, despite the low

differentiation of the Batrachophrynus gene pool for the allozymes we studied (comparable

to studies on bird allozymes), we do not doubt the specific status of both taxa which we

consider sister species.

Source : MNHN, Paris

64 ALYTES 13 (2)

Batrachophrynus Telmatobius

brachy. macro. jelskii rimac culeus

EST2 (90)

PEP1 (83)

MDH1 (125)

LDH1 (33)

LDH1 (100)

HDH1

PEP2.1

6PGD

PGMi

Fig. 5. — Proposal for the phylogenetic relationships among five species of Andean Telmatobiinae,

based on the presence (dot) and absence (circle), respectively, of alleles at different allozyme loci.

The cluster formed by Telmatobius samples agrees with the morphometric assignment

of populations to the species T. jelskii and T. rimac. The genetic distance between the

conspecific samples correlates with the corresponding geographical distance of localities.

Finally, the low genetic distance between T. jelskii and T. rimac as well as the absence of

diagnostic fixed alleles indicates a close phylogenetic relationship.

The evaluation of fixed alleles and of presence/absence of rare alleles supports the

same tree structure as the quantitative treatment of allele frequencies. Moreover, it allows

for a proposal on the relationships of T. culeus with the central Peruvian taxa (fig. 5). Yet,

the large geographical distance between the current distribution ranges of T. jelskii and T.

rimac on one hand, and T. culeus on the other, indicates a long period of independent

evolution. Nevertheless, the fixed allele at the LDHI1 locus clearly suggests that T. culeus

is a member of the same Te/matobius lineage as are the central Peruvian species. This

shared character state distinguishes Te/matobius from Batrachophrynus and favours the

hypothesis of an early separation put forward by LyNCH (1978). The geographically

distant T. culeus genetically and morphometrically resembles the Batrachophrynus species

more than the neighboring central Peruvian Te/matobius species do. This may indicate that

Batrachophrynus species represent the remnants of an early invasion into the central

Peruvian Andes, whereas the Telmatobius species reached this region during a second, later

invasion. The limited present range of distribution of Batrachophrynus, and the suspicious

Source : MNHN, Paris

SINSCH & JURASKE 65

absence of streams occupied by both B. brachydactylus and a stream-inhabiting

Telmatobius species, suggest that Telmatobius is competitively superior to Batrachophry-

nus. Future field work in the contact zone between B. brachydactylus and T. jelskii in the

streams of the Junin area should reveal whether the two can coexist in the same stream.

Our study supports the taxonomic distinction of the central Peruvian Telmatobiinae

into two genera Batrachophrynus and Telmatobius (PETERS, 1873). The geographical

distribution of Batrachophrynus and Telmatobius species indicates the result of competition

between early and late invaders of this region rather than phylogenetic proximity.

However, the genetic distance between the members of these genera is low. Data from

additional Telmatobius species and from an appropriate outgroup (Alsodes, considered as

sister taxon of Telmatobius) are needed for a final decision on the relationships between

Batrachophrynus and Telmatobius.

RESUMEN

Se revisan tres hipôtesis sobre’el origen filogenético de los Telmatobiinae del Perû

central: (1) el antepasado comün de las dos especies de Batrachophrynus se separé del stock

de Telmatobius antes de su diferenciaciôn en las especies recientes; (2) B. macrostomus se

separé primero, B. brachydactylus mâs tarde del stock de Telmatobius; (3) la separaciôn del

antepasado comün de Batrachophrynus del stock de Telmatobius ocurrié despues de su

diferenciacion. Alocimas demuestran claramente que B. brachydactylus y B. macrostomus

son familiares muy cercanos, y que T. culeus del sur del Perû es relacionado mâs cercano

con T. jelskii y T. rimac del Perû central que con las especies de Batrachophrynus. En

conclusién, todas las pruebas disponibles apoyan la primera hipôtesis.

ACKNOWLEDGEMENTS

We are grateful to Lic. J. CoRDOVA, curator of the herpetological section of the Museo de la

Historia Natural de la Universidad Nacional de San Marcos, Lima, permitting us access to the

Telmatobiinae of the local collection. M. ANTIGNANI, J. ICOCHEA and A. SALAS helped us collect frogs

in the field, W. BÔHME generously permitted us to take blood samples from his T. culeus specimen,

and B. NiLow provided technical assistance. The paper benefited from the comments of E. BALLETTO,

W. R. HEYER and of an anonymous referee on an earlier draft.

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CavaLLi-SrorZaA, L. L. & EDWARDS, A. W. F., 1967. — Phylogenetic analysis: models and estimation

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Cr, J. M., 1986. — Speciation and adaptive radiation in Andean Telmatobius frogs. In: F.

VUILLEUMIER & M. MONASTERIO (eds.), High altitude tropical geogeography, New York, Oxford

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Source : MNHN, Paris

66 ALYTES 13 (2)

DuELLMAN, W. E., 1979. — The herpetofauna of the Andes: patterns of distribution, origin,

differentiation and present communities. In: W. E. DUELLMAN (ed.), The South American

herpetofauna: its origin, evolution and dispersal, Monogr. Mus. nat. Hist. Univ. Kansas, 7:

371-459,

Kansas, 21: 1-372.

FELSENSTEIN, J., 1985. — Confidence limits in phylogenies: an approach using the bootstrap.

Evolution, 39: 783-791.

Frost, D. E., 1985. — Amphibian species of the world. A taxonomic and geographic reference.

Lawrence, Allen Press and Assoc. Syst. Collections: 1-732.

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Cambridge, 3: 273-278.

LAURENT, R. F., 1983. — Heterogenidad del género Batrachophrynus Peters (Leptodactylidae). Acta

zool. lill., 37: 107-113.

LAviLLA, E. O., 1988. — Lower Telmatobiinae (Anura: Leptodactylidae): generic diagnoses based on

larval characters. Occ. Pap. Mus. nat. Hist. Univ. Kansas, 124: 1-19.

LyNCH, J. D., 1978. — A re-assessment of the telmatobiine leptodactylid frogs of Patagonia. Occ.

Pap. Mus. nat. Hist. Univ. Kansas, T2: 1-57.

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NÔLLER, H. G., 1959. — Eine einfache Technik der Blutentnahme beim Frosch. Pflügers Arch.

Physiol., 269: 98-100.

PASTEUR, N., PASTEUR, G., BONHOMME, F., CATALAN, J. & BRITTON-DAVIDIAN, J., 1988. - Practical

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REYNOLDS, J. B., WEIR, B. S. & COCHERHAM, C. C., 1983. — Estimation of coancestry coefficient:

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Corresponding editors: Alain Dusois & W. Ronald HEYER.

Source : MNHN, Paris

Alytes, 1995, 13 (2): 67-76. 67

Advertisement, aggressive,

and possible seismic signals

of the frog Leptodactylus syphax

(Amphibia, Leptodactylidae)

Adäo J. CARDOsO * & W. Ronald HEYER **

* Departamento de Zoologia, Universidade Estadual de Campinas,

Caixa Postal 6109, CEP 13081-970 Campinas, SP, Brasil

** Department of Vertebrate Zoology, National Museum of Natural History,

Smithsonian Institution, Washington, DC 20560, U.S.A.

Advertisement calls from three geographically isolated populations of

Leptodactylus syphax are remarkably similar. Aggressive calls of L. syphax

sound to the human ear very different from the advertisement calls, although

the basic structural components are similar. À male L. syphax responded with

aggressive calls to a playback of his own advertisement call. The same male

responded to playbacks of his agressive calls with increased rate of aggres-

sive calls and foot pounding behavior. The foot pounding produced audible

clicks and, by its nature, seismic signals. This is the second known instance of

Leptodactylus species producing seismic signals, each produced differently,

however. It is not known whether L. syphax interprets the seismic signals.

Seismic signalling in frogs may be much more common than currently

believed.

INTRODUCTION

Leptodactylus syphax Bokermann, 1969 is restricted to rocky granitic outcroppings

and is known from a few disjunct, widely separated, localities in Brazil (fig. 1). The first

author recently recorded calls from three of these disjunct populations. We analyze the

advertisement calls of the frogs from these three populations to determine whether there

is any significant variation among them. At one locality, the first author was fortunate to

observe and record Leptodactylus syphax aggressive calls and foot-pounding behavior. The

foot-pounding may involve seismic communication, previously reported for the first time

in frogs by Lewis & NARINS (1985). We describe and comment on all of these calls and

behaviors.

Source : MNHN, Paris

68 ALYTES 13 (2)

Fig. 1. — Known distribution of Leptodactylus syphax in South America. Triangles: sites from which

recordings are analysed in this paper (westernmost triangle: Barra do Bugres, Mato Grosso

State; northernmost triangle: Säo Raimundo Nonato, Piaui State; southernmost triangle:

Alpinépolis, Minas Gerais State). Square: site from previously published recording by W. C. À.

BOKERMANN (Chapada dos Guimaräes, Mato Grosso State). Dots: other known localities (note

that southernmost dot, the locality of Serra do Espinhaço, Minas Gerais State, was incorrectly

placed in northeastern Brazil in HEYER, 1970, fig. 21).

Source : MNHN, Paris

CARDOSO & HEYER 69

METHODS AND MATERIALS

Recordings were made using a Uher Report 4000 reel-to-reel tape recorder. The

recording information is:

(1) Tape ASN/AJC (Archivo Sonoro Neotropical/Adäo J. CARDoso) 13, cut 6, Brazil,

Minas Gerais State, Alpinépolis, Fazenda Salto; no voucher specimen; recorded by A. J.

CARDOS0; 11 October 1981; 21.00 hours; 22°C air temperature. Ten advertisement calls are

analyzed from this individual.

(2) Tape ASN/AJC 84, cut 2, Brazil, Mato Grosso State, Barra do Bugres, Reserva

Biolôgica Serra das Araras; no voucher specimen; recorded by A. J. CARDOsO; 19

November 1988; 20.30 hours; 26°C air temperature. Sixteen advertisement calls are

analyzed from this individual.

(3) Tape ASN/AJC 101, cut 2, Brazil, Piaui State, Säo Raimundo Nonato, Parna,

Serra da Capivara, localidade Caldeiräo; voucher specimen ZUEC 8829 (Universidade

Estadual de Campinas); recorded by A. J. CARDoso; 4 March 1990; 20.00 hours; 27.5°C

air temperature. Ten advertisement calls, 9 aggressive calls, and 1 foot pounding are

analyzed from this individual.

The recordings were analyzed with “Canary” software from the Cornell Laboratory

of Ornithology on a Macintosh Ilci computer. The sampling rate used to convert the

analogue signals to digital format was 22,254.5 Hz with 8-bit precision. Filter bandwidths

of 353 Hz and frame lengths of 256 points were used for both audiospectrogram and

spectrogram analyses.

Call terminology follows that defined in HEYER et al. (1990).

RESULTS

ALPINOPOLIS DATA

Three individuals were calling at the recording site. The calling males were separated

from each other by a distance greater than 100 m, far from water, in an area characterized

by large rocks, among which crevices were abundant. The individual recorded was calling

near one of these crevices, into which it fled after being approached to within about 10 m.

Only the advertisement call was recorded; no playback was presented to the frog.

The advertisement call (fig. 2 A), is given at an average rate of 0.8 per second. Call

duration ranges from 59 to 64 ms. The call is frequency modulated with a rapid rise time;

the broadcast frequency range sweeps from 390 to 2110 Hz with maximum broadcast

intensity between 1310 and 1330 Hz. The call is strongly partially pulsed, typically with

3 almost completely defined pulses. Harmonics are present (not particularly visible on

fig. 2 À, but spectrogram analyses of calls [not shown] indicate their presence).

Source : MNHN, Paris

70 ALYTES 13 (2)

6 7 4 Ë ë

|°

le ge je ë Sr sl

Frequency (kHz)

6 -

lo) ; ë

j É

4 7 F É

j £

f # É.

î:

DE FAR Re PR

1 2 3

Time (s)

Fig. 2. — Audiospectrograms of advertisement calls of Leptodactylus syphax. Upper figure (A)

recorded from Alpinépolis; middle figure (B) recorded from Barra do Bugres; lower figure (C)

recorded from Säo Raimundo do Nonato.

0 ne

Source : MNHN, Paris

CaARDOSsO & HEYER 71

BARRA DO BUGRES DATA

A single individual was calling from this locality, which was characterized by large

sheets of rock and no water systems obvious in the area. Advertisement calls only were

recorded. The individual stopped calling when approached within about 20 m and did not

respond to playback of its call when broadcast from near the calling site.

The advertisement call (fig. 2 B) is given at an average rate of 1.5 per second. Call

duration ranges from 53 to 60 ms. The call is frequency modulated with a rapid rise time;

the broadcast frequency range sweeps from 380 to 2300 Hz with maximum broadcast

intensity between 1800 and 1850 Hz. The call is partially pulsed, with 3 to 5 weakly defined

pulses (the recording has a low frequency component making precise determination of the

number of pulses difficult). Harmonics are present (energy analyses of calls [not shown]

indicate their presence).

SAO RAIMUNDO NONATO DATA

Two individuals were calling at the site, about 200 m from each other. The individual

recorded was calling from an extensive horizontal rock fissure. The opening of the crevice

was about 30 cm high and 5 m long. The crevice extended about 4 m into the rock wall,

on the face of a waterfall, which at the time had little running water.

Initially the advertisement call was recorded without the monitor on, such that the

frog did not hear its own voice. At this time, the calling frog was about 4 m from the

microphone. After the initial recording was made, the monitor button was engaged and the

frog began to hear its own voice from the tape recorder speaker. Immediately after hearing

its own voice, the individual started to emit aggressive calls intermixed with advertisement

calls and jumped to within about 2 m of the tape recorder. The emissions were given at

a very variable rate with considerable irregularity in the bursts of call types. After a while

of recording under these conditions, a section of tape with a series of aggressive calls was

played back to the frog. On hearing the playback of these aggressive calls, the frog

increased its rate of aggressive calls and beat its forelimbs on the ground, thereby causing

the foot pounding sound. The tape recorder was then stopped, recording was begun anew

with the monitor engaged, such that the frog could hear its own sounds from the tape

recorder speaker, including the foot pounding sounds. Soon thereafter, the frog jumped to

the side of the tape recorder, emitted various sounds, and abruptly stopped calling. The

frog was then collected to serve as a voucher for the recordings.

The advertisement call (figs. 2 C, 3), is given at an average rate of 1.2 per second. Call

duration ranges from 56 to 64 ms. The call is frequency modulated with a rapid rise time;

the broadcast frequency sweeps from 390 to 2060 Hz with maximum broadcast intensity

between 1640 and 1680 Hz. The call is either composed of two extremely well-defined

pulses (almost notes), the first with about 3 weakly defined partial pulses, or composed of

about 4 weakly defined partial pulses. Harmonics are present.

Source : MNHN, Paris

Amplitude

Magnitude (dB)

ALYTES 13 (2)

r r — r

20 40 60 80 100

Time (ms)

1695

-80

-100

3390

5085 6780

-120 1 0

8475

-140 NV A ha

r r r r T

2 4 6 8 10

Frequency (kHz)

Fig. 3. — Wave form and energy analysis of advertisement call of Leptodactylus syphax recorded from

Säo Raimundo do Nonato. Upper figure shows wave form (the pulse above the 80 milliseconds

label is interpreted as microphone ringing, not a part of the call); bracket above wave form

indicates portion of call used for spectrogram analysis of lower figure.

Source : MNHN, Paris

CaRDOsO & HEYER 73

The aggressive call (figs. 4, 5) is given at an average rate of 0.7 per second. Call

duration ranges from 162 to 206 ms. The call is frequency modulated in a complex fashion.

There is an initial low intensity fast rise in frequency, the fundamental rising from 220-

480 Hz to 920-1010 Hz, followed by a falling frequency, steeper initially, the fundamental

from about 920-1010 Hz falling to 310-520 Hz. The call is partially pulsed with about

3 weakly defined pulses. There are at least 3 well-defined harmonics which have about as

much broadcast energy as the fundamental.

There is no apparent transition in call when a male switches from advertisement to

aggressive calls (and vice versa); the male utters either one kind or the other (fig. 5).

The pounding of the front foot results in a faint, but audible click (fig. 4), that has

most energy at about 1800 Hz. Foot pounding, by its nature, produces seismic signals as

well.

DISCUSSION

The calls from the three isolated populations of Leptodactylus syphax studied are

remarkably similar, differing only in details that might be expected to occur among

individuals from a single population (see GERHARDT, 1988, for a general discussion and

RyaAN, 1980, for a specific example analyzing fundamental frequency). The calls reported

here are also similar to the call from Chapada dos Guimaräes, Mato Grosso State,

recorded by Werner C. A. BOKERMANN, previously reported (HEYER, 1979), with one

exception. The previously analyzed recording from Chapada dos Guimarâes gave very

little indication of harmonic structure. However, harmonic structure is evident in the wave

forms of the calls analyzed herein (e. g., fig. 3, above), and the spectrogram analysis

indicates the presence of at least 3 harmonics in addition to the fundamental frequency

(fig. 3, below). These differences in harmonic expression may be due to differences of

recording and analytic equipment rather than actual call differences.

The modest differences in advertisement calls among the geographic samples analyzed

are somewhat surprising. We do not know whether these similarities may be due to recent

isolation of the presently disjunct populations of L. syphax or due to selection for

stabilization of the advertisement call among all populations.

The advertisement and aggressive calls are very different sounding (and appearing

when analyzed) calls. They sound as though they were calls of two different species of

frogs. The advertisement and aggressive calls differ in duration and mode of frequency

modulation. The calls do share the characteristics of being frequency modulated, having

overlapping broadcast frequencies, and having harmonic structure. These similarities

suggest that the same physical structural complex is involved in producing both calls and

the differences are produced by a combination of behavioral:controls regulating the

duration of the call and manipulating tension of the laryngeal musculature which causes

changes in the tension of the vocal cords resulting in differences of the physical structure

of the emissions. These behavioral changes are not trivial, however. Lewis & NARINS

(1985) reported similar results for Leptodactylus albilabris. While the advertisement call of

Source : MNHN, Paris

74 ALYTES 13 (2)

Frequency (kHz)

Time (s)

Fig. 4. — Audiospectrogram of two aggressive calls and foot-pounding of Leptodactylus syphax

recorded from Säo Raimundo Nonato. The arrow indicates the foot-pounding sound.

3,2

Ë À es

; 7

0 abat at

Time (s)

Fig. 5. — Audiospectrogram of continuous recording of a male Leptodactylus syphax from Säo

Raimundo Nonato, uttering aggressive calls (A) followed immediately by advertisement calls (B)

with no intermediate call structure between the two call types.

Source : MNHN, Paris

CARDOSO & HEYER 75

L. albilabris is short and rises from 1000 to 2300 Hz, the male-male interaction call is

longer and descends from 2300 to 1000 Hz. Perhaps this pattern of frequency modulation

reversal and time differences in advertisement and aggressive calls is common to all

Leptodactylus species with rising whistle-like advertisement calls.

Lewis & NaRINs (1985) and NaRINs (1990) reported that Leptodactylus albilabris

produces and is capable of receiving and interpreting seismic signals. Lewis & NARINS

(1985) speculated that the different arrival times of the simultaneously produced seismic

and airborne waves could provide a temporal clue to the distance from the source and

could be used to help males establish and maintain territories. NARINS speculated that L.

albilabris might be able to integrate the seismic and air-borne advertisement calls “to better

communicate when high-level background noise obscures the acoustic channel” (NARINS,

1990: 273). This could also pertain to L. syphax, as the habitats they call from have noisy

waterfalls during rainy periods.

The mechanism for producing seismic signals in L. albilabris was reported to be the

rapidly expanding vocal sac contacting the ground. Leptodactylus syphax produce seismic

signals by beating their forefeet on the ground. In contrast to the seismic signals of L.

albilabris, which are not audible to the human ear, the foot pounding of L. syphax is

weakly audible to the human ear, and is certainly within the frequency range of the

advertisement call of L. syphax. We assume the audible nature of the foot pounding of L.

syphax is possibly due to the presence of horny spines on the inner thumb of the male. In

contrast to L. albilabris, the seismic signals made by L. syphax are not produced

simultaneously with advertisement or aggressive calls. We do not know whether L. syphax

is processing the air-borne click portion of the foot-pounding, the seismic signals, neither,

or both. However, we report here the second known instance in frogs producing seismic

signals, both within the genus Leptodactylus, but by very different methods. Obviously,

this foot-pounding behavior of L. syphax merits further study as well as detailed study of

other frogs to determine whether seismic signalling is much more common than currently

believed.

ACKNOWLEDGMENTS

A short-term visitor grant to the first author from the Office of Fellowships and Grants,

Smithsonian Institution, provided the initial collaboration that led to this paper. Additional support

to the first author was from the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico

(CNPq) and to the second author from the Museu de Zoologia da Universidade de Säo Paulo, the

Neotropical Lowlands Research Program of the Smithsonian Institution’s International Environ-

mental Sciences Program, and the Director’s Office, National Museum of Natural History.

Ronald I. CROMBIE, Smithsonian Institution, reviewed the manuscript for us. Two anonymous

reviewers suggested several improvements.

Source : MNHN, Paris

76 ALYTES 13 (2)

LITERATURE CITED

GerHarDT, H. C., 1988. — Acoustic properties used in call recognition by frogs and toads. Jn: B.

FRiTzsCH, M. J. RyAN, W. WILCZYNSKI, T. E. HETHERINGTON & W. WALKOWIAK (eds.), The

evolution of the amphibian auditory system, New York, John Wiley & Sons: 455-483.

HEYER, W. R., 1979. — Systematics of the pentadactylus species group of the frog genus Leptodactylus

(Amphibia: Leptodactylidae). Smithsonian Contrib. Zool., 301: 1-43.

HEYER, W. R., RAND, A. S., CRUZ, C. A. G., PEIXOTO, O. L. & NELSON, C. E., 1990. — Frogs of

Boracéia. Arg. Zool., 31: 231-410.

Lewis, E. R. & Nains, P. M., 1985. — Do frogs communicate with seismic signals? Science, 227:

187-189.

NaRINS, P. M., 1990. — Seismic communication in anuran amphibians. BioScience, 40: 268-274.

RYAN, M. J., 1980. — Female mate choice in a Neotropical frog. Science, 209: 523-525.

Corresponding editor: Walter HôDL.

Source : MNHN, Paris

Alytes, 1995, 13 (2): 77-80. 77

Captive maintenance of adults

and juveniles of the genus Triturus

during the terrestrial phase

Verina WAIGHTS

Department of Biology, The Open University, Walton Hall, Milton Keynes, MK7 6AA, United Kingdom

Two methods for maintaining newts during the terrestrial phase, such that

regular recapture could be effected, were compared. The first method, which

has been used previously by other workers for plethodontids, led to failure to

thrive and absence of breeding condition during the following spring. The

second method, which sought to mimic the natural terrestrial habitat of

species of the genus Triturus, énabled efts to reach sexual maturity in one year

and adults to subsequently come into breeding condition.

INTRODUCTION

The development of a method to maintain newts in the terrestrial phase, whereby

individuals can easily be recaptured, is important for several reasons. Little is known about

growth of juveniles or adults during this phase in the wild, mainly due to the difficulty of

locating individuals (GRIFFITHS, 1984). Enhanced feeding of efts may lead to sexual

maturity within one year which may assist reproductive studies on captive populations

(BAKER, 1988; ELEBERT, 1991). However, raising juveniles to sexual maturity in one year

may not be desirable for programmes where the newts are to be re-introduced into the

wild, as it may impose unnatural selection pressures.

Newts, captured aquatically, can rapidly lose breeding condition. Males often react to

the stress of capture by rapid regression of their crests or tail filaments and both sexes may

cease courtship behaviour. VERRELL (1982) found that these reactions to stress in the male

could be overcome in Notophthalmus viridescens by enhanced feeding but I found this

regime to be unsuccessful for both Triturus montandoni and Triturus helveticus (unpub-

lished data). Maintenance of adults during the terrestrial phase, enabling them to come

into breeding condition the following spring, may facilitate studies of courtship behaviour

in the laboratory.

Workers in North America have maintained plethodontids terrestrially, which

enabled them to observe courtship behaviour for eight months of the year (SEVER &

Houcx, 1985). This method has also been successfully used to rear Notophthalmus efts

(VERRELL, 1983).

Below, two methods are described, which were investigated for maintaining Triturus

species during the terrestrial phase; the first method is based on that used for

plethodontids and the second method is an attempt to mimic the natural terrestrial habitat

of Triturus species.

Source : MNHN, Paris

78 ALYTES 13 (2)

METHODS AND RESULTS

FIRST METHOD

Adult Triturus helveticus were collected from a pond in Bedfordshire, England, and

allowed to mate in tanks (30 cm x 60 cm x 38 cm high, size 1) at 14°C and on a natural

photoperiod. The resulting larvae were fed on Daphnia and Tubifex. At metamorphosis,

the efts were transferred to small, round glass dishes (10 cm diameter X 4 cm high), lined

with moist paper towel, containing crumpled paper for them to hide under and covered

with ‘“parafilm” to maintain the humidity and to prevent their escape. Each week the

moist towel was renewed. The efts were kept at a density of six to a dish and maintained

at 10-12°C with a 12L:12D photoperiod and fed on fruit flies (Drosophila) ad libitum.

After initially emerging from the water onto rocks, the adult newts were observed to

re-enter the water several times before their skins reverted to the terrestrial velvety

condition. They were then transferred to transparent boxes (17 cm x 31 cm x 9 cm high),

at a density of four to a box and maintained under the same conditions as the efts.

The efts failed to thrive; they grew fast initially but then remained at a small size and

subsequently died. None reached sexual maturity. The adults tended to lose weight during

the terrestrial phase, appearing very thin and dark skinned; none came into breeding

condition the following spring.

SECOND METHOD

Triturus alpestris adults were collected in France in 1989, in breeding condition. They

were housed in aquaria (size 1) and maintained at 12°C on an artificial photoperiod that

replicated the natural photoperiod (condition A). The resulting larvae were fed on Daphnia

and Tubifex. Ten larvae metamorphosed during late summer.

Triturus montandoni adults were collected in Poland in 1990, in breeding condition.

However, when they reached the laboratory, the males’ tail filaments had regressed and

they failed to court. The females, which had been ovipositing prior to capture, failed to

deposit any more eggs. Within a few weeks the adults left the water, via rocks emerging

from it, and were transferred to a terrarium.

The efts and adults were kept in terraria consisting of transparent plastic tanks

(21 cm x 40 cm x 25 cm high) covered with a plastic lid containing mesh over the air

holes. A layer of earth, 5 cm deep, was put in the bottom of the tank. Dry leaf litter, which

contained small invertebrates (wood lice, ants, beetles, etc.), was placed on top of the earth

until the tank was half-full, followed by several large stones and some pieces of bark for

the newts to hide under. The terraria were maintained so that the soil base was always

moist and the leaf litter dry. Each terrarium contained up to ten efts or six adults, which

were fed on Drosophila (flies and maggots) and white worms (Enchytraeus albidus) ad

libitum, and maintained at 19-23°C with a natural photoperiod. One corner of the

terrarium was used to maintain the white worms, which were replenished regularly.

Source : MNHN, Paris

WAIGHTS 79

In December, the terraria were transferred to condition À, to simulate winter, and fed

as above. In February, when Triturus vulgaris were migrating to the ponds locally, the

adult T. montandoni were transferred to aquaria (size 1), filled to a depth of 15 cm, and

kept in an unheated shed with a natural photoperiod. The T. alpestris efts had also thrived

and their skins now appeared damp, so they were transferred to tanks identical to the

above. The newts were placed on bricks above the water level.

In the terraria, the newts were often found clustered together under a piece of bark

and the efts were found inside the curled leaves. The adults were also found buried in the

soil, however they could still be seen foraging during the late afternoon and evening.

Within a few days of transfer, the adult T. montandoni became aquatic. Subsequently,

five of the six came into breeding condition, courted and reproduced. Six juvenile T.

alpestris became aquatic during the first day. They also came into breeding condition but,

as all the juveniles were female, no courtship was observed.

DISCUSSION

Looking at the results described above, it appears that the regime used in North

America so successfully for plethodontids is inappropriate for Triturus spp. In the first

method, although the absolute humidity may change as the week progresses due to the

moist towel drying out, the humidity is uniform throughout the box. Little is known about

the humidity preferences of terrestrial newts; therefore, the humidity achieved in the box

may be unsuitable. The diet in this method is also very uniform. SMITH (1951) described

the terrestrial diet of newts as consisting of worms, slugs, snails and insects; therefore, it

is possible that the newts are being deprived of essential nutrients when fed solely on a

single species of insect.

The second method was equally successful for maintaining both efts and adults. The

terrarium was set up so that a humidity gradient existed within it and the newts appeared

able to find an appropriate microhabitat. The diet was more varied, the newts were fed on

Drosophila (flies and maggots), white worms and any invertebrates in the soil and leaf

litter. This diet may be closer to that found in the wild and is therefore more likely

to supply all the nutrients necessary for spermatogenesis and somatic growth. This

method also gave the newts a period of “summer” terrestrial conditions which has been

shown to be a requirement for complete spermatogenesis to occur (SAEZ et al., 1992). The

lack of this “summer” period in the first method may have contributed to the decline of

the newts and may indicate that efts also require these higher temperatures for

development.

Development of a method that facilitates recapture in the terrestrial phase may allow

investigation of differential feeding and growth rates during this phase. Age accounts for

only a small proportion of the total variance in adult body size (HALLIDAY & VERRELL,

1988) and therefore a study of juvenile growth during this phase may help to elucidate

further the variation in subsequent adult body size. BAKER (1992) has shown that crest

height is related to body condition in Triturus cristatus. The above method will enable a

Source : MNHN, Paris

80 ALYTES 13 (2)

study to be undertaken to investigate the relationship between growth and body condition

in the terrestrial phase to the development of secondary sexual characteristics and

subsequent reproductive success in the aquatic phase.

RÉSUMÉ

Deux méthodes pour le maintien en captivité de tritons pendant leur phase terrestre

sont décrites et comparées. La première méthode, qui a été déjà utilisée avec succès avec

des Pléthodontidés, a abouti à un échec en ce qui concerne la croissance des animaux et

leur aptitude à se reproduire au printemps suivant. La deuxième méthode, qui cherche à

reproduire l'habitat terrestre naturel des espèces du genre Triturus, a permis aux jeunes

tritons d’atteindre la maturité sexuelle en un an et aux adultes de retrouver leur condition

reproductive au printemps.

ACKNOWLEDGEMENTS

I thank Herbert MACGREGOR and Leigh GiLLETT for providing the animals, Fred TOATES and

Alain Duois for translating the Résumé into French, John BAKER and Tim HALLIDAY for much

helpful advice and Günter GOLLMANN and two anonymous referees for assistance in revising the

manuscript. This work was supported by the Open University.

LITERATURE CITED

BAKER, J. M. R., 1988. — Maintenance and breeding of Triturus karelini. Brit. herp. Soc. Bull., 25:

42-44.

ee 1992. — Body condition and tail height in great crested newts, Triturus cristatus. Anim. Behav.,

43: 157-159.

ELEBERT, E., 1991. — Precocious newts. Brit. herp. Soc. Bull., 35: 17-19.

GRIFRITHS, R. A., 1984. — Seasonal behaviour and intrahabitat movements in an urban population

of smooth newts, Triturus vulgaris (Amphibia: Salamandridae). J. Zool., Lond., 203: 241-251.

HALLIDAY, T. R. & VERRELL, P. A., 1988. — Body size and age in amphibians and reptiles. J. Herp.,

22 (3): 253-265.

SAEZ, F. J., FRAILE, B., PAZ DE MIGUEL, M. & PANIAGUA, R., 1992. — Effects of low temperature

on testicular cells in the marbled newt, Triturus marmoratus (Caudata, Salamandridae). Herp.

J., 2: 125-132.

Sever, D. M. & Houck, L., 1985. — Spermatophore formation in Desmognathus ochrophaeus

(Amphibia: Plethodontidae). Copeia, 1985: 394-402.

SmirH, M., 1951. — The British amphibians and reptiles. London, Collins: 1-318.

VERRELL, P. A., 1982. — A note on the maintenance of the red spotted newt in captivity. Brit. herp.

Soc. Bull., 5: 28-29.

ee 1983. — A note on the breeding of the red-spotted newt in captivity. Brit. herp. Soc. Bull. 6:

48-49.

Corresponding editor: Günter GOLLMANN.

Eu) Source : MNHN, Paris

AINTTES

International Journal of Batrachology

published by ISSCA

EDITORIAL BOARD FOR 1995

Chief Editor: Alain Durois (Laboratoire des Reptiles et Amphibiens, Muséum national d'Histoire

naturelle, 25 rue Cuvier, 75005 Paris, France). )

Depurx Editor: Janalee P. CaLDwELL (Oklahoma Museum of Natural History, University of

Oklahoma, Norman, Oklahoma 73019, U.S.A.).

Editorial Board: Ronald G. ALTIG (Mississippi State University, U.S.A.}: Emilio BALLETTO (Torino,

Italy): Alain COLLENOT (Paris, France), Günter GOLLMANN (Wien, Austria), Tim HALLIDAY

(Milton Keynes, United Kingdom); W. Ronald HEvER (Washington, U.S.A.); Walter HôDL

(Wien, Austria): Pierre JoLY (Lyon, France): Masafumi Maïsur (Kyoto, Japan); Jaime

E. PÉFaUR (Mérida, Venezuela): J. Dale RoBERTs (Perth, Australia); Ulrich Sixsc (Koblenz,

Germany): Marvalee H. WakE (Berkeley, U.S.A.).

Technical Editorial Team (Paris, France): Alain DuBoIs (texts); Roger BoUR (tables); Annemarie

OuLer (figures)

Index Editors: Annemarie OHLER (Paris, France); Stephen J. RicHaRDs (Townsville, Australia).

GUIDE FOR AUTHORS

Alytes publishes original papers in English, French or Spanish, in any discipline dealing with

amphibians. Beside articles and notes reporting results of original research, consideration is given for

publication to synthetic review articles, book reviews, comments and replies, and to papers based

upon original high quality illustrations (such as color or black and white photographs), showing

beautiful or rare species, interesting behaviors, etc.

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be typewritten or printed double-spaced on one side of the paper. The manuscript should be organized

as follows: English abstract, introduction, material and methods, results, discussion, conclusion,

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not exceed 16 x 24 cm. The size of the lettering should ensure its legibility after reduction. The

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References in the text are to be written in capital letters (SOMEONE, 1948: So & So, 1987;

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BOURRET, R., 1942. - Les batraciens de l'Indochine. Hanoï, Institut Océanographique de l'Indochine:

ix + 1-547, pl. I-IV.

Grar, J.-D. & POLLs PELAZ, M., 1989. - Evolutionary genetics of the Rana esculenta complex. In:

R. M. DaAWLEY & J. P. BOGART (eds.), Evolution and ecology of unisexual vertebrates, Albany, The

New York State Museum: 289-302.

INGER, R. F., Voris, H. K. & Voris, H. H., 1974. - Genetic variation and population ecology of some

Southeast Asian frogs of the genera Bufo and Rana. Biochem. Genet., 12: 121-145.

Manuscripts should be submitted in triplicate either to Alain DuBoIs (address above) if dealing

with amphibian morphology, systematics, biogeography, evolution, genetics or developmental

biology, or to Janalee P. CALDWELL (address above) if dealing with amphibian population genetics,

ecology. ethology or life history. Acceptance for publication will be decided by the editors following

review by at least two referees.

If possible, after acceptance, a copy of the final manuscript on a floppy disk (3 % or 5 W)

should be sent to the Chief Editor. We welcome the following formats of text processing: (1)

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No page charges are requested from the author(s), but the publication of color photographs is

charged. For each published paper, 25 free reprints are offered by Alyies to the author(s). Additional

reprints may be purchased.

Published with the support of AALRAM

(Association des Amis du Laboratoire des Reptiles et Amphibiens

du Muséum National d'Histoire Naturelle, Paris, France).

Directeur de la Publication: Alain DuRoïs.

Numéro de Commission Paritaire: 64851.

Alytes, 1995, 13 (2): 45-80.

Contents

E. O. LaviLA & P. ERGUETA S.

Una nueva especie de Telmatobius (Anura, Leptodactylidae)

de la ceja de montaña de La Paz (Bolivia)

Ulrich SNsCH & Norbert JURASKE

Reassessment of central Peruvian Telmatobiinae

(genera Batrachophrynus and Telmatobius).

11. Allozymes and phylogenetic relationships ........................ 52-66

Adäo J. CARDOsO & W. Ronald HEYER

Advertisement, aggressive, and possible seismic signals

of the frog Leptodactylus syphax (Amphibia, Leptodactylidae) ....... 67-76

Verina WAIGHTS

Captive maintenance of adults and juveniles

of the genus Zriturus during the terrestrial phase ... 77-80

Alytes is printed on acid-free paper.

Alytes is indexed in Biosis, Cambridge Scientific Abstracts, Current Awarèness in Biological

Sciences, Pascal, Referativny Zhurnal and The Zoological Record.

Imprimerie F. Paillart, Abbeville, France.

Dépôt légal: 2°" trimestre 1995.

Source : MNHN, Paris