Sex Determination of Adult Humboldt Penguins Using
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J. Field Ornithol., 68(1):102-112 SEX DETERMINATION OF ADULT HUMBO•T PENGUINS USING MORPHOMETRIC CHARACTERS CAgLOS B. ZAVALAGA 1 AND ROSANA PAREDES ProyectoConservaci6n de Punta SanJuan WildlifeConservation Society ParqueJosg de Acosta187 Lima 17 Perd Abstract.-•TheHumboldt Penguin (Spheniscushumboldti) is one of the most endangered and least-studiedspecies of penguins.Studies of its breedingecology are needed to develop managementplans for its conservation,and sex determination is essentialto measurecertain reproductiveparameters. Humboldt Penguins,like other speciesof penguins,showed sexual sizedimorphism, with malesbeing heavierand larger than females.One discriminantfunc- tion wasderived from externalmeasurements of 297wild adult HumboldtPenguins of known sex (146 femalesand 151 males) at Punta SanJuan, Peru. The sex of 95% of these birds could be correctlydetermined using the discriminantfunction D• -- -38.98 + 3.16(WH) + 3.69(BL) where WH is width of the head and BL is bill length. In addition, the sexof 91% of the birdswere correctly classified by meansof the bill lengthusing the univariatefunction D,= -6.31 + BL. In both equations,if D > 0 the bird was classifiedas male, if D < 0 it was a female, and if D = 0, the sex could not be identified. The discriminant function D 1 wasnot accurateto sexa groupof captiveHumboldt Penguins (71% successfullyclassified) becausethere were differencesin sizeof the head betweenwild and captivebirds. However, weobtained 83% of casessuccessfully classified using the functionD 2 as bill lengthwas similar betweenwild and captivebirds. The discriminantfunction may not be applicableto other wild and captivepopulations of Humboldt Penguins. DETERMINACION DEL SEXO EN ADULTOS DE SPHENISCUS HUMBOLDTI UTILIZANDO CARACTERISTIC• MORFOM•TRICAS Sinopsis.--Dentrode las 17 especiesde pingtinos,el de Humboldt (Spheniscushumboldti) es uno de los m•s amenazadosy menos estudiados.Para 11evara cabo planes de manejo dirigidosa su conservaci6n,son necesariosestudios sobre su ecologiareproductiva, siendo la determinaci6ndel sexoesencial para medir algunosde susparimetros reproductivos. Los pingtinos de Humboldt son sexualmentedim6rficos en susmedidas corporales, siendo los machosmils pesados y grandesque las hembras.Una funci6n discriminantefue obtenidaa partir de lasmedidas de 297 adultosde sexoconocido (146 hembrasy 151machos) en Punta SanJuan, Per6. E1sexo de estasaves pudo ser identificadocorrectamente en un 95% de los casosutilizando la funci6n discriminante D 1 = -38.98 + 3.16 (WH) + 3.69 (BL), donde WH esel anchode la cabezay BL la longituddel pico. De igual manerase identific6 el sexo del 91% de los pingtinos por medio de la ecuaci6nunivariada D2 = -6.31 + BL. Para ambasecuaciones, si D > 0 el ave fue clasificadacomo macho,si D < 0 como hembray si D = 0, el ave no era clasificada. La funci6n discriminante D• no fue tan exitosa en la determinaci6ndel sexode pingtinos cautivos(s61o el 71% de los casosfue correctamente clasificado)debido a que estasaves exhibieron cabezas mils grandes que aquellasmedidas en el campo.Sin embargo,el sexodel 83% de lospinguinos pudo ser identificado utilizando la ecuacionD,debido a que la longituddel pico fue similarentre los ping/iinossilvestres y los cautivos.La funci6n discriminantedesarrollada no necesariamentepudiera aplicarsea otraspoblaciones de pingtinos de Humboldt ya seasilvestres o cautivos. The Humboldt Penguin (Spheniscushumboldti) is one of the most threatened and least-studiedspecies of penguins (Boersma 1991, Hays Currentaddress: Apartado Postal 180606, Lima 18, Perd. 102 Vol.68, No. I SexingHumboldt Penguins [ 103 1984). Becauseof a sharp decline in the wild population (Hays 1984), studiesof their ecologyand behavior are needed for developingeffective conservationplans. To achieve most of these studies,the ability to deter- mine the sex of individuals is essentialfor investigatingsex-specific be- havior (Davis 1988) or measuring certain reproductive parameters (CCAMLR 1994). Determining sex of captiveHumboldt Penguinsin zoos is also important in order to maximize successfulreproduction in captive birds (Cheney 1990, McGill and Perkins 1993, Steveson1993). Humboldt Penguins,like most seabirdspecies, lack plumage characters by which sexesmay be recognized. However, small differences in mor- phometric parameters reveal sufficient dimorphism to distinguishsexes (Murphy 1936:452,Scholten 1987). Charactersused to determinesex in penguinsinclude vent measurements(Boersma and Davies1987), sexual behavior (Davis 1988, Edgington 1989, Scholten 1992), morphometric differences in mated pairs (Edgington 1989) and temporal attendance during incubation (Kerry et al. 1993). Some of thesemethods have been used successfullyin the wild and in captivity,but only on reproductive individualsduring early parts of the breeding season.Cloacal examination has been used in wild Adfilie Penguins (Pygoscelisadeliae,, Lishman 1985, Sladen 1978), ChinstrapPenguins, (Pygoscelis antarctica; Lishman 1985) and captive Humboldt Penguins (Yamazaki et al. 1994). However, this technique requires trained researchers and specialized equipment to complete with minimal stressto the animal. Other methods,such as chro- mosome analysis,hormone analysis,and laparoscopyare effective but time-consumingand expensive (Edgington 1989). Sex determination by discriminant analysisof external measurements has been used successfullyin the wild in severalpenguin speciessuch as MagellanicPenguin (Spheniscusmagellanicus, Scolaro et al. 1983), Yellow- eyed Penguin (Megadyptesantipodes, Darby and Seddon 1990), all pygos- celid penguins (Amat et al. 1993, Kerry et al. 1992, Williams 1990), Little Blue Penguin (Eudyptulaminor novaehollandiae,Gales 1988), and others. Discriminant analysis uses morphometric differences between known male and female birds to calculate a function that predicts the sex of unknown individuals. This technique is reliable, practical, fast, inexpen- sive,non-invasive, and can be used outside of the breeding season. The objectivesof this studywere (1) to determine discriminantfunc- tion(s) using external measurementswith which to determine the sex of wild adult Humboldt Penguins at Punta San Juan, Peru and (2) to test the applicabilityof these functions on captiveHumboldt Penguins. MATERIALS AND METHODS Measurementsof wild penguins.--BetweenSeptember 1992 and June 1993, the bodies of 223 dead adult Humboldt Penguinswere recovered from the port of San Juan de Marcona, 3 km north of Punta San Juan (15ø22'S,75ø12'W), a 54-ha guano bird reserveon the southerncoast of Peru. This reserve holds one of the largest populations of Humboldt Penguinsin the country (Hays 1984). The birds were accidentallycaught 104] c. B. Zavalagaand R. Paredes j. FieldOrnithol. Winter 1997 and drowned in the nets of small boatsin the courseof normal fishing operationsbased at Punta SanJuan along 50-60 km of the coast (within 5 km offshore).Thus, the penguinscaught may originatefrom Punta San Juan as well as from neighboring colonies such as Sombrerillo, San Ni- colils, and San Fernando. Immediatelyafter collection,carcasses were weighedto the nearest100 g with a 10-kg Pesolaspring balance. Head and bill were measuredwith vernier calipersto the nearest0.1 mm and the flipper wasmeasured with a ruler to the nearest 0.1 cm. The measurementsincluded: total length of the head (LH) from supraoccipitalto the tip of the bill; width of the head (WH) in a crevicejust posteriorto the bulge behind the eyes;bill length (BL) from the edge of implantationof feathersto the tip of the culmen;bill depth (BD) measureddorso-ventrally at the nostrilsand flip- per length (FL), maximumflattened chord, from the humero-radialjoint to the tip of the flipper (Fig. 1). The sex of each individual was deter- mined by gonad examination after dissection. Discriminant functions were derived from the measurements taken from the 223 carcasses(analysis sample). These data were analyzedby stepwisediscriminant function analysiswith the SPSS/PC + 4.0.1 statis- tical package (Norusis 1990). The Wilks' lambda ratio was used as a cri- terion for variable selection.All other statisticalanalysis were carried out using the SYSTAT5.0 statisticalpackage (Wilkinson 1991). To test the accuracyof our results,we measured74 breedingwild pen- guins of known sex (validation sample). These animalswere banded and measuredbetween 1992 and 1994 at Punta San Juan as part of a long- term studyof the breeding ecologyof this species.The sex of thesebirds was identified from their position during copulation. Body masswas excluded from the discriminantanalysis because there was a significantamount of seasonalvariability (CV = 13.6%). Also, the majority of dead penguins were wet and their lungs were full of water (due to drowning) at the time of weighing, which would bias this mea- surement(Table 1). These penguinswere 4.8% heavierthan a sampleof 288 living penguins of known sex (the sex of these individualswas deter- mined from the discriminantfunction reported below) weighedat the sametime at Punta SanJuan (males:• -- 4711.06 g, SE -- 31.55, range -- 3450-6000, n = 165; females:• = 4047.39 g, SE -- 35.15, range = 2950-5400, n = 123; pooled:• = 4427.62 g, SE = 30.42, range = 2950- 6000, n -- 288). These differences were significant (z-test, z = 98, P < 0.05). All measurementswere normallydistributed (Kolmogorov-Smirnov one sample test, P > 0.05 in all tests)with some overlap between sexes.In order to determine a discriminant function, we established the same cri- FIGURE1. Variablesmeasured