Overcoming environmental and morphological constraints: Egg size and pelvic kinesis in the smallest tortoise, Homo...

See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/236981903 Overcoming environmental and morphological constraints: Egg size and pelvic kinesis in the smallest tortoise, Homo... Article in Canadian Journal of Zoology October 2005 DOI: 10.1139/z05-132 CITATIONS 28 READS 100 3 authors: Margaretha Hofmeyr University of the Western Cape 87 PUBLICATIONS 664 CITATIONS Brian Henen 116 PUBLICATIONS 1,379 CITATIONS SEE PROFILE SEE PROFILE Victor Loehr Homopus Research Foundation 51 PUBLICATIONS 306 CITATIONS SEE PROFILE Some of the authors of this publication are also working on these related projects: Tortoise ecology and conservation View project Tortoise and terrapin phylogeography View project All content following this page was uploaded by Margaretha Hofmeyr on 21 July 2014. The user has requested enhancement of the downloaded file.

1343 Overcoming environmental and morphological constraints: egg size and pelvic kinesis in the smallest tortoise, Homopus signatus M.D. Hofmeyr, B.T. Henen, and V.J.T. Loehr Abstract: The small tortoises of southern Africa include the only testudinid taxa that produce single-egg clutches. This group includes the world s smallest tortoise, Homopus signatus (Gmelin, 1789), which inhabits a harsh, arid environment. Climate and body size may influence reproductive output, so we hypothesized that the east west aridity gradient in southern Africa affects egg and clutch size of the small indigenous tortoises, and that the morphology of H. signatus constrains egg size, preventing the formation of optimal eggs. Here we show that aridity and unpredictable rainfall determine which of these tortoise taxa produce single-egg clutches. Taxa in less predictable environments produce larger eggs relative to body size than do taxa in more predictable environments. Homopus signatus produces the largest egg relative to body size, probably to enhance offspring survival in its harsh environment. Body size, pelvic aperture size, and the narrow anal gap of H. signatus appear to constrain egg size. Despite these constraints, females produce rigidshelled eggs larger than the pelvic canal and use pelvic kinesis to pass eggs at oviposition; both features are unknown in other chelonians and emphasize the selective advantage of large eggs to H. signatus. Résumé : Les petites tortues du sud de l Afrique contiennent les seuls taxons de testudinidés qui produisent une seule couvée. Ce groupe comprend les plus petites tortues du monde, Homopus signatus (Gmelin, 1789), qui habitent des milieux rudes et arides. Comme le climat et la taille corporelle influencent vraisemblablement le rendement reproductif, nous avons émis l hypothèse que le gradient est ouest d aridité dans le sud de l Afrique affecte la taille des oeufs et celle des couvées des petites tortues indigènes et que la morphologie d H. signatus limite la taille des oeufs, empêchant ainsi la formation d oeufs optimaux. Nous démontrons que l aridité et les précipitations imprévisibles déterminent lesquels de ces taxons produisent une seule couvée. Les taxons qui vivent dans les milieux moins prévisibles produisent des oeufs plus gros relativement à leur taille que les taxons des milieux plus prévisibles. Homopus signatus produit les oeufs les plus gros, compte tenu de sa taille, probablement afin d améliorer la survie des ses rejetons dans un environnement rude. La taille du corps, la dimension de l ouverture pelvienne et l étroitesse du passage anal semblent restreindre la taille des oeufs. Malgré ces contraintes, les femelles produisent des oeufs à coquille rigide plus grands que leur canal pelvien et utilisent la cinétique du pelvis pour faire passer les oeufs lors de la ponte. Ces caractéristiques sont inconnues chez les autres chéloniens, ce qui souligne l avantage sélectif des gros oeufs chez H. signatus. [Traduit par la Rédaction] Hofmeyr et al. 1352 Introduction Natural selection typically favours resource allocations that optimize maternal and offspring fitness. Successful allocation schemes vary considerably, however, as attested by the large variety of life-history strategies (Congdon et al. 1982; Stearns 1992; Kuchling 1999). Variation in life histories can be attributed, largely, to the multitude of selective pressures on allocation schemes, including maternal and environmental effects. Often, life histories demonstrate allocation trade-offs between offspring number and size (Elgar and Heaphy 1989; Iverson and Smith 1993), and large offspring may have increased fitness (Janzen 1993; but see Congdon et al. 1999). Females should allocate enough resources to produce viable offspring. Yet, if allocating additional resources to individual offspring negligibly enhances offspring fitness, females should produce more, not larger, offspring. Under these conditions, optimal egg size theory states that, if resource availability changes, egg size should vary little in comparison to clutch size (Smith and Fretwell 1974; Congdon 1989; Congdon and Gibbons 1990). Body size constrains offspring number and size in small chelonians, and morphological constraints may prevent females from producing optimal eggs (Congdon and Gibbons Received 3 May 2005. Accepted 1 September 2005. Published on the NRC Research Press Web site at http://cjz.nrc.ca on 18 October 2005. M.D. Hofmeyr 1 and B.T. Henen. Chelonian Biodiversity and Conservation Southern Africa, Department of Biodiversity and Conservation Biology, University of the Western Cape, PB X17, Bellville 7535, South Africa. V.J.T. Loehr. Chelonian Biodiversity and Conservation Southern Africa, Department of Biodiversity and Conservation Biology, University of the Western Cape, PB X17, Bellville 7535, South Africa, and Homopus Research Foundation, Nipkowplein 24, 3402 EC IJsselstein, the Netherlands. 1 Corresponding author (e-mail: mdhofmeyr@uwc.ac.za). Can. J. Zool. 83: 1343 1352 (2005) doi: 10.1139/Z05-132

1344 Can. J. Zool. Vol. 83, 2005 1987). In some small-bodied chelonians, egg width and pelvic canal width correlate similarly to body size, and the width of the large females eggs exceeds the pelvic canal width of the small females. These results suggest that pelvic canal size can constrain egg width (Congdon and Gibbons 1987). Additionally, sexual dimorphisms in the pelvic apertures of some tortoise species (Long and Rose 1989) imply a selective advantage of large pelvic apertures for females. The posterior shell opening between the plastron and carapace (anal gap) may also constrain egg size in turtles and tortoises (Rose and Judd 1991; Clark et al. 2001), although shell kinesis or shell hinges may ameliorate this constraint. Environmental variation and unpredictability can significantly influence resource allocation schemes and life histories (Congdon et al. 1982; Stearns 1992; Kuchling 1999). Chelonians at high latitudes often produce larger but fewer clutches than do taxa at lower latitudes, a trend probably related to the short favourable nesting and incubation season at high latitudes (Iverson et al. 1993; Kuchling 1999, among others). The bet-hedging concept addresses the risk of producing too many offspring at one time (Murphy 1968). Since fitness in unpredictable environments can be increased by producing fewer than the maximum number of offspring (Philippi and Seger 1989), the bet-hedging strategy results in smaller reproductive allocations that occur more frequently (Louw and Seely 1982; Lovegrove 1993). Southern Africa s rich tortoise fauna developed from one or more waves of Eurasian immigrations during the Oligocene and Miocene. By the early Miocene, three African lineages could be distinguished: Stigmochelys, Centrochelys, and the Ethiopian endemics (Lapparent de Broin 2000). Each of these lineages diversified into several species through the Miocene and Pliocene (Lapparent de Broin 2000 and references therein) when a strong east west aridity gradient developed in southern Africa (Partridge 1997). The extant genera of the Ethiopian endemics are small to mediumsized tortoises and four of these genera, together with the Malagasy genus Pyxis, contain the only tortoise taxa that produce single-egg clutches (Ernst and Barbour 1989; Boycott and Bourquin 2000). Evaluating reproductive allocations in the single-egg clutching taxa and their congeners (i.e., the small endemics ) may elucidate evolutionary forces that influenced the life histories of these tortoises. For 13 of these taxa, we assessed the effects of body size and environment on egg size, clutch size, and clutch volume. When considering environmental effects, we tested for effects of latitude, rainfall, and rainfall predictability. The southern African endemics include the world s smallest tortoise, the speckled padloper (Homopus signatus (Gmelin, 1789)) (Boycott and Bourquin 2000). Homopus signatus produces one egg at a time (Loehr et al. 2004) and inhabits rocky terrain in the harsh arid environs of southern Africa (Boycott and Bourquin 2000). The combination of a small body size and arid climate poses a great physiological challenge, especially for females who must allocate resources to eggs while ensuring their own survival. Singleegg clutching tortoises cannot trade-off egg size with clutch size, but they may still trade-off egg size with clutch frequency. Although females should benefit from producing optimal eggs, a larger egg may enhance hatchling fitness in an arid environment. In a small species such as H. signatus, morphological constraints may prevent females from producing optimal eggs. To evaluate egg-size optimality and morphological constraints in the smallest tortoise, we related egg size to body size, pelvic canal size, and the anal gap of live and preserved H. signatus. We also evaluated sexual dimorphisms that may facilitate the production of large eggs. Understanding the reproductive allocations of small tortoises in arid environments should provide new insights into the evolution of life histories. Materials and methods Body size and egg size We used published and unpublished life-history data on tortoise genera that contain single-egg clutching taxa. These taxa include Homopus femoralis Boulenger, 1888; Homopus areolatus (Thunberg 1787); Homopus boulengeri Duerden, 1906; Homopus sp.; Homopus signatus; Psammobates geometricus (L., 1758); Psammobates tentorius tentorius (Bell, 1828); Psammobates tentorius verroxii (Smith, 1839); Psammobates tentorius trimeni (Boulenger, 1886); Psammobates oculiferus (Kuhl, 1820); Chersina angulata (Schweigger, 1812); Malacochersus tornieri (Siebenrock, 1903); and Pyxis arachnoides Bell, 1827. We will refer to this group as the small endemics. Tortoise variables included female averages for body mass (BM), straight carapace length (SCL), egg length (EL), egg width (EW), and clutch size (CS, midrange values). Because egg mass data were not available for all taxa, we used EL and EW measures to calculate egg volume: EV = π EL EW 2 /6 (Coleman 1991). BM but not body volume (BV) data were available for all taxa, so we calculated relative egg volume (REV) as the ratio of EV to BM. Environmental variables included latitude, annual rainfall (AR), and rainfall predictability (RP = 100 the coefficient of variation of AR, %). For our analyses of environmental parameters, we used midrange values and minimum values for each taxon. Rainfall and RP values were not available for M. tornieri and P. arachnoides, so we evaluated these effects for 11 of the 13 taxa. Homopus signatus morphometrics We studied the reproduction of wild Homopus signatus signatus females during their breeding season over 2 consecutive years at Springbok, South Africa (August to October 2000 and 2001; see Loehr et al. 2004). Females were captured opportunistically and promptly weighed (±0.1 g; Soehnle Ultra digital balance) before using vernier callipers (±0.01 cm) to measure SCL (nuchal to supracaudal), plastron length (PL), maximum shell height (SH, typically at the third vertebral scute), and maximum shell width (SW). For this analysis, we considered only the gravid females (N = 31). We supplemented measurements on wild specimens with measurements from preserved H. signatus females (N = 20) and males (N = 15) from the South African Museum. For preserved specimens, we measured SCL, SW, SH, posterior shell height (SHP, at the anterior edge of the fifth vertebral scute, just dorsal to the iliosacral joint), and anal gap (AG, the midline distance between the supracaudal scute and su-

Hofmeyr et al. 1345 Fig. 1. X-ray radiograph of a gravid Homopus signatus female. The large, oval-shaped egg, orientated at an angle to the vertebral column (at the midline), has to be expelled through the pelvic canal. The pelvic canal is visible below, between the partially withdrawn hind limbs. Brightness ( 20) and contrast (+20) of the digital image were enhanced with Adobe Photoshop version 6.0. magnification (Graham and Petokas 1989). For object-tofilm distances, one half of SHP was used to correct PW measurements and one half of SW was used to correct PH measurements. Three museum specimens were gravid and the elevation of an egg from the plastron, as seen in lateral radiographs, was used to correct egg size magnification in the dorsoventral radiographs. In dorsoventral radiographs, the perpendicular distance of an egg to the line representing the lateral-view film cassette was used to correct egg size in the lateral radiographs. Corrected egg sizes from the two views were very similar (P > 0.5, coefficient of variation 1.0%), validating our method. The correction for radiographic magnification of wild tortoise measurements was made using the ratio of true PL to radiographic PL. This method gave similar results to the Graham and Petokas (1989) method (see Loehr et al. 2004). The corrected EL and EW measures were used to estimate EV. Since the long axis of an egg may not be parallel to the film plane, radiographic measures of EL may be considered unreliable. However, vertical inclination of the egg should be small in H. signatus. Homopus signatus are dorsoventrally flattened and produce a very wide egg. The low SH and large egg should restrict vertical inclination of the egg. The nearly identical EL results from lateral and dorsoventral radiographs confirmed that egg inclination had negligible effects. ture of the anal scutes). For wild and preserved tortoises, shell volume (SV) was estimated using a modified formula for an ellipsoid (SV = π/6 SCL SH SW, cm 3 ; Loehr et al. 2004). Homopus signatus egg and pelvis size We radiographed wild H. signatus females dorsoventrally (Fig. 1; 50 kvp (kilovolt peak) for 0.25 s at 50 ma) to determine the width of the pelvic canal (PW), and to measure EL and EW of gravid females (Loehr et al. 2004). Museum specimens were radiographed dorsoventrally and laterally (35 45 kvp, focal distance = 800 mm) to determine PW and pelvic canal height (PH) of males and females, and egg dimensions (EL and EW) of gravid females. As a proxy for measuring ilial inclination, we measured the distance of the ilial heads and acetabula to the distal edge of the supracaudal scute in the museum radiographs. For lateral radiographs, the tortoise midline was positioned parallel to the film, with the widest part of the carapace touching the film cassette. We measured PW at the largest gap between the ilia, which bow outward midway along their dorsoventral length. PH was measured from the floor (puboischium near the ilia) to the ceiling (iliosacral joint). For radiographs of museum specimens, we used focal distance and object-to-film distances to correct for radiographic Statistics and allometry We report results as mean, 95% confidence interval, and sample size (N), and considered statistical tests significant at P < 0.05. Data were first tested for normality and equal variance, and if transformations failed, we used nonparametric tests (e.g., Spearman s rank order correlations, r S ). Simple linear regressions were used to evaluate scaling phenomena and reproductive relationships to environmental variables. We also evaluated scaling phenomena on a log log basis to test for isometric (slope = 1) or allometric (slope 1) relationships; log log slopes were compared with 1.0 using Student s t tests. All of our findings were allometric with log log slopes < 1 (i.e., the variables did not increase in direct proportion to one another). We used forward stepwise regressions, confirmed with backward stepwise regressions, to assess which independent variables should be included in multiple regression models. Analysis of covariance (ANCOVA) was used to assess group differences when dependent variables correlated with body size. In ANCOVA, we tested for equal slopes and if slopes did not differ, we tested for equal elevations of regression lines (Zar 1999). We wanted to use multiple regression analysis to compensate for the effects of body size while assessing the effects of female body condition on egg size. However, multicollinearity between BM and BV caused us to use residual analyses (Clark et al. 2001). Residuals of BM to BV provide a measure of body condition. Residuals of EV on BV remove the effect of body size on egg size. We scaled EV residuals against BM residuals to determine if female body condition influenced egg size. Before committing resources to eggs at ovulation, egg precursors represent nutrient reserves to females (Henen 1997), so we report results relating EV residu-

1346 Can. J. Zool. Vol. 83, 2005 Table 1. Female reproductive and environmental parameters of African tortoise genera that include single-egg clutching taxa. Species BM (g) SCL (mm) EV (cm 3 ) REV (%) CS AR (mm) RP (%) Lat ( S) Homopus femoralis 393.0 a 135.0 14.2 a,b 3.62 1 3 c 250 750 65 80 28 33 Homopus areolatus 229.4 d 107.4 8.94 d 3.90 1 3 d 250 1000 70 85 31 35 Homopus boulengeri 177.1 a 102.0 10.79 e,f 6.09 1 c 100 500 60 80 30 34 Homopus sp. 234.8 g 108.8 15.17 g 6.46 h 1 g <100 30 50 26.5 28 Homopus signatus 151.4 i 92.6 11.34 i 7.49 1 i <100 250 50 75 29 33 Psammobates geometricus 473.9 j 127.0 9.32 j 1.97 1 5 j 500 750 75 85 33.5 34.5 Psammobates tentorius tentorius 508.1 k 130.5 10.65 k 2.10 1 3 k 150 500 65 85 31 34 Psammobates tentorius verroxii 331.1 a 121.0 10.56 l 3.19 1 2 c,g 100 500 40 75 23 32 Psammobates tentorius trimeni 250.6 a 113.2 10.56 l 4.21 1 c,m <100 200 30 75 27 31 Psammobates oculiferus 317.9 a 119.0 16.43 n 5.17 1 n 100 500 30 80 17 30 Chersina angulata 907.8 j 170.1 24.22 j 2.67 h 1 j,m <100 800 50 85 28 35 Malacochersus tornieri 382.5 o 150.7 15.96 o 4.17 h 1 m,o na na na Pyxis arachnoides 301.2 j 116.1 13.46 p 4.47 1 p na na na Note: Data is organized by genus and relative egg volume; sources are indicated by letter superscripts. BM is average body mass; SCL is average straight carapace length and has the same source as BM; EV is average egg volume calculated from egg width and length; REV is relative egg volume (EV/BM); CS is clutch size; AR is annual rainfall over the range of the taxon (Lovegrove 1993; Schulze 1997); RP, rainfall predictability = 100 the coefficient of variation of AR (derived from Lovegrove 1993; Schulze 1997); Lat is latitude (Swingland and Klemens 1989; Boycott and Bourquin 2000). a Western Cape Nature Conservation Board herpetological records (accessed 2003). b Branch 1999. c Boycott and Bourquin 2000. d B.T. Henen, M.D. Hofmeyr, and G. Kuchling, unpublished data. e Loveridge and Williams 1957. f Haagner 1990. g A. Schleicher, unpublished data. h Egg measures from captive animals. i Loehr et al. 2004. j M.D. Hofmeyr, B.T. Henen, and G. Kuchling, unpublished data. k T.E.J. Leuteritz, unpublished data. l Branch 1989. m Species occasionally lay two-egg clutches. n Rall 1990. o F.A.C. Schmidt, unpublished data. p Durrell et al. 1989. als to BM residuals inclusive of egg mass. We obtained similar results when egg mass was excluded. Results Egg and clutch size Although BM among the small endemics varied sixfold, EV varied less than threefold (Table 1). Body size had an allometric effect on EV and clutch volume (for BM and SCL: all r 2 > 0.37, F [1,11] > 6.7, and P < 0.025). The effect of body size was stronger when we considered only the single-egg clutching taxa (for BM and SCL: all r 2 > 0.74, F [1,6] > 18.0, and P < 0.01). In contrast, body size did not influence CS in the small endemics (P = 0.32). EV of the single-egg clutching taxa and their congeners correlated inversely with CS (r S = 0.61, P = 0.024, N = 13). The inverse relationship between EV and CS remained after removing the effect of body size on egg size via multiple regression (EV = 0.017BM 3.9CS + 12.8, r 2 = 0.82, F [2,10] = 22.5, P < 0.001; BM: P < 0.001; CS: P < 0.005). Environmental parameters also affected reproduction. Stepwise regression for egg size eliminated latitude and AR from the model, and retained BM (P < 0.005) and RP (P < 0.05). Using multiple regressions to account for body size, RP was inversely related to EV (r 2 = 0.71, F [2,8] = 10.0, P < 0.01; BM: P < 0.005; RP: P < 0.05) and EW (r 2 = 0.72, F [2,8] = 10.4, P < 0.01; BM: P < 0.005; RP: P < 0.05). The regression for EL on body size and RP was not significant overall (P = 0.08) or for BM (P = 0.09), but was significant for RP (P < 0.05). For clutch volume, stepwise regression removed latitude and RP but retained AR and BM (clutch volume = 0.019AR + 0.014BM + 6.8, r 2 = 0.71, P < 0.01; AR: P < 0.05; BM: P = 0.07). Stepwise regression for CS removed body size, latitude, and RP from the model but retained AR (P < 0.01). CS increased with increasing AR (CS = 0.0026AR + 0.61, r 2 = 0.57, F [1,9] = 11.9, P < 0.01) and the effect was stronger when considering the lowest AR values instead of midrange values, over each taxon s distribution (r 2 = 0.86, F [1,9] = 55.1, P < 0.001). Although the effect of RP on CS was not as strong as the effect of AR on CS, CS increased significantly with RP (CS = 0.039RP 1.0, r 2 = 0.51, F [1,9] = 9.3, P < 0.05). The effect of RP on CS was stronger when we considered the lowest RP instead of midrange values, over each taxon s distribution (r 2 = 0.60, F [1,9] = 13.3, P < 0.01). Egg size and pelvic constraint in Homopus signatus Homopus signatus females produce large eggs (Fig. 1); EL and EW were 37% and 27%, respectively, of SCL. The relative EV averaged 7.5% of BM, exceeding relative egg size for the other taxa (Table 1). Egg size varied with maternal size in wild H. signatus, with EV and EW scaling

Hofmeyr et al. 1347 Fig. 2. Relationship of egg length (EL), egg width (EW), and egg volume (EV) to straight carapace length for wild H. signatus. The relationships were significant only for EV and EW (EV = 0.19SCL 6.6, r 2 = 0.58; EW = 0.18SCL + 8.3, r 2 = 0.61; both F [1,29] > 39.5 and P < 0.001; N = 31). Egg size (mm or cm ) 3 45 35 25 15 EW 5 80 90 100 110 120 Carapace length (mm) Fig. 3. Relationship (log log) of egg width (EW, circles) and pelvic canal width (PW, triangles) to straight carapace length for wild H. signatus (log EW = 0.69 log SCL + 0.036, r 2 = 0.62, F [1,29] = 46.5, P < 0.001; log PW = 0.70 log SCL 0.018, r 2 = 0.41, F [1,29] = 20.2, P < 0.001; N = 31). Regression slopes were similar (t 58 = 0.043, P > 0.5) but elevations differed (t 59 = 6.82, P < 0.001). Egg or pelvis width (log, mm) 1.50 1.45 1.40 1.35 1.30 allometrically with SCL (Fig. 2). The coefficient of variation was similar for EL (5.5%) and EW (6.4%), yet EL did not scale to carapace length. Although H. signatus produce rigid-shelled, nonpliable eggs, 93.5% of the eggs were wider than their pelvic canals. Eggs exceeded the corresponding PWs by 2.1 ± 0.47 mm or 9.5% (range = 4.0% to 20.7%; N = 31). EW and PW scaled similarly to SCL (Fig. 3) and the slopes of the log log regressions were significantly less than 1 (P < 0.05). The regressions had similar slopes, but the elevation was significantly higher for EW. The widest part of the pelvic canal was approximately 45% (16% 107%, N = 20) wider than the narrow gap between the ilial heads. EW scaled allometrically with PW (Fig. 4) and the regression line approaches the isometric line for PW. EW PW EL EV 1.25 1.90 1.95 2.00 2.05 2.10 Carapace length (log, mm) Fig. 4. Relationship of egg width (EW) to pelvic canal width (PW) for wild H. signatus females (EW = 0.63PW + 10.5, r 2 = 0.52, F [1,29] = 31.6, P < 0.001; N = 31). For females with wide pelvic girdles, EW approached the isometric line for PW. Egg and pelvis width (mm) 30 27 24 21 EW PW isometric line 18 19 21 23 25 27 Pelvic canal width (mm) Fig. 5. Relationship of (A) pelvic canal width (PW, circles), pelvic canal height (PH, triangles) and (B) anal gap (AG, inverted triangles) to straight carapace length for museum specimens of H. signatus males (open symbols; N = 15) and females (solid symbols; N = 20). For males, PW and PH scaled allometrically (PW = 0.12SCL + 6.1, r 2 = 0.34, F [1,13] = 6.80, P < 0.05; PH = 0.098SCL + 7.6, r 2 = 0.43, F [1,13] = 9.84, P < 0.01). For females, only PW scaled with SCL (PW = 0.24SCL + 0.081, r 2 = 0.30, F [1,18] = 7.71, P < 0.05). AG scaled to SCL for males (AG = 0.35SCL 15.0, r 2 = 0.61, P < 0.001) and females (AG = 0.32SCL 17.2, r 2 = 0.23, P < 0.05). Pelvis width or height (mm) Anal gap (mm) 28 24 20 16 20 16 12 8 A B Male Female 4 65 70 75 80 85 90 95 100 105 Carapace length (mm) Sexual dimorphism The relationship of PW to SCL did not differ between wild and museum females (ANCOVA: similar slopes and elevations, P > 0.5), validating our measurements on museum specimens. Homopus signatus females were significantly

1348 Can. J. Zool. Vol. 83, 2005 larger than males (Fig. 5; SCL: females = 92.1 ± 1.7, N = 20; males = 78.6 ± 2.4, N = 15; t 33 = 9.97, P < 0.001) and pelvic girdles showed pronounced sexual dimorphisms. The width of the pelvic canal did not differ from PH for either sex (P > 0.8; Fig. 5A). For males, PW and PH scaled with SCL, while in females, PW but not PH scaled with SCL. Pelvic canals were considerably wider for females than for males (ANCOVA elevations: t 32 = 5.11, P < 0.001). The AG of males and females scaled on SCL (Fig. 5B). AGs were similar for sexes (males: 12.7 ± 4.15 mm, N = 15; females: 12.3 ± 5.30 mm, N = 20), but after correcting for SCL, males had larger AGs (ANCOVA elevations: t 32 = 3.82, P < 0.001). The AG of females (range 7.1 17.8 mm) was much narrower than EW of wild tortoises (24.9 ± 0.54 mm, N = 35, range = 22.2 28.6 mm) and museum specimens (23.5 26.0 mm, N = 3). For H. signatus females, the narrowest egg was 25% 210% (4.6 15.1 mm) wider than any measured AG. The posterior portion of the shell, at the pelvic girdle (SHP), was significantly higher in females than in males (ANCOVA: slopes similar (t 31 = 0.26, P > 0.5), but elevations differed (t 32 = 3.55, P < 0.002)). In the side-view radiographs of H. signatus, the ilia of male pelves were more vertically oriented than were female ilia. Although both ilialacetabular and ilial-supracaudal distances scaled to SCL for males and females, only the ilial-supracaudal distance differed between sexes; male distances were larger (ANCOVA elevations: t 32 = 4.43, P < 0.001). Body size and condition Eggs represent a relatively large proportion of body size for small females compared with that for large females (range = 6.2% 10.6% of SV; EV = 0.040SV + 5.7, r 2 = 0.56, F [1,29] = 37.0, P < 0.001; slope significantly smaller than one for the log log regression, P < 0.05). Additionally, H. signatus females in better condition, i.e., BM relative to BV, tend to produce larger eggs (Fig. 6). The increased EV was effected through increased EW (EW residual: r 2 > 0.26, F [1,29] = 10.3 and P < 0.005); EL did not scale to female volume (P = 0.11). Discussion Fig. 6. Correlation of relative egg size (egg volume (EV) to body volume residual) to body condition as relative body mass (body mass (BM) to body volume residual) in wild H. signatus females (EV residual = 0.073(BM residual) 6.3 10 6, r 2 = 0.33, F [1,29] = 14.4, P < 0.001; N = 31). Egg volume residual (cm ) 3 4 3 2 1 0-1 -2-3 -4-30 -20-10 0 10 20 30 Body mass residual (g) Body size and egg size The single-egg clutching tortoises and their congeners are small to medium sized, and the larger tortoises tend to make larger eggs than do the smaller tortoises. However, the smaller tortoises invest proportionately more resources into an egg than do the larger tortoises. Within these taxa, there is a trade-off between egg size and CS. That is, relative to body size, taxa that invest more resources in individual offspring produce fewer offspring per clutch. A similar trade-off between egg size and number has been shown at the generic and family levels for other chelonians (Elgar and Heaphy 1989; Rowe 1994). Of the few nontestudinid chelonians that produce single-egg clutches, most inhabit humid or tropical regions (e.g., Platemys platycephala (Schneider, 1792), Rhinoclemmys punctularia (Daudin, 1802), and Kinosternon angustipons Legler, 1965; Ernst and Barbour 1989), not arid and unpredictable habitats. These species have medium to small body sizes, so a relatively large egg may be linked partially to a small body size. The smallest turtle that produces single-egg clutches, K. angustipons, also produces a relatively large egg (SCL = 120 mm, EV = 10.1 cm 3 ; Ernst and Barbour 1989). Within the small endemics, body size had no direct effect on CS. Factors other than body size thus determine which of these taxa produce single-egg clutches. CS seems determined by AR (Table 1); females in mesic habitats produce smaller eggs and larger clutches than do females from arid zones. Primary production decreases in parallel with the east west aridity gradient in southern Africa (Schulze 1997) and high productivity probably enhances the viability of small eggs. Small eggs often lead to small hatchlings that should feed, grow, and survive more readily among the high productivity of mesic habitats (e.g., contrast H. areolatus with more arid zone Homopus spp.). In contrast, the xeric species seem to benefit from larger eggs; in arid regions, larger eggs may be more viable than small eggs. Arid regions have less predictable rainfall (Table 1; Louw and Seely 1982), so arid zone females may benefit by producing large eggs with nutrient reserves that facilitate hatchling survival during the first unpredictable months. Large eggs and large hatchlings have relatively small surface to volume ratios, improving their ability to counter desiccation. However, the consistent clutch volumes among congeners (ca. 6% 8% and 4% 6% of BM for Homopus spp. and Psammobates spp., respectively) suggest there is a constraint on the allocations per clutch. This type of constraint may force arid zone species to limit CS in exchange for producing large eggs. In the smallest tortoise, H. signatus, the large size of females compared with males and the effect of maternal size on egg size indicate that body size is an important constraint in female reproduction. The large female size may facilitate digging nests deep enough to minimize physical constraints, e.g., high temperatures or evaporation rates, on incubating eggs; females dig nests under vegetation with eggs covered by about 4 cm of sand (wild and captive H. signatus; Loehr 1999 and unpublished data). The eggs of H. signatus are more than twice the EV of the smallest turtles, Sternotherus depressus Tinkle and Webb, 1955 (SCL = 115 mm; EV =

Hofmeyr et al. 1349 4.5 cm 3 ) and Glyptemys muhlenbergii (Schoepff, 1801) (SCL = 115 mm; EV = 3.5 cm 3 ; Ernst and Barbour 1989), but these turtles produce more than one egg per clutch. Among the small endemics, H. signatus eggs are large on both absolute and relative scales (Table 1), emphasizing the importance of large eggs to female H. signatus. The large range in H. signatus egg size suggests that small females eggs must confer a greater fitness advantage than deferring reproduction until females attain a larger body size. However, small females must pay a disproportionate price; their eggs represent a relatively large investment in comparison to investments for large H. signatus females. Good body condition may help smaller females compensate for their limiting size. If more resources (e.g., body reserves) are available, H. signatus females of all sizes will make bigger eggs. Morphological constraints on egg size Egg size in wild H. signatus scaled allometrically with maternal size, but female size appears to constrain EW more than EL. EW and PW in H. signatus scaled similarly with SCL, indicating that the size of the pelvic canal aperture constrains egg size, as has been indicated for some other small chelonians (e.g., Chrysemys picta (Schneider, 1783) and Deirochelys reticularia (Latreille in Sonnini and Latreille, 1801); Congdon and Gibbons 1987). In H. signatus, the width of the pelvic canal did not differ from PH. The eggs are essentially circular in cross section, so there was no asymmetry in how the pelvic canal might limit the passing egg. Unlike the regressions for C. picta and D. reticularia, however,ewinh. signatus exceeded the width of the pelvic canal. This is the first record of a chelonian with eggs wider than the pelvic canal. PW may constrain EW in C. picta and D. reticularia (Congdon and Gibbons 1987), but their canal width was reported as the shortest distance between the ilia. The eggs of C. picta and D. reticularia were 4% 12% narrower than the narrowest ilial gap (Congdon and Gibbons 1987). We measured PW at the largest gap between the ilia. The outward bow of the ilia accommodates the widest part of the egg at oviposition. For H. signatus, EW exceeded the widest opening between the ilia, which was approximately 45% wider than the gap between the ilia heads. Consequently, the pelvic constraint in H. signatus appears much greater than pelvic constraints reported for other chelonians. Homopus signatus make rigid-shelled, nonpliable eggs, which must pass through the pelvic canal and AG at oviposition. To pass an egg, the small AG of H. signatus females must open proportionately more than their pelvic canal opens. Although our H. signatus findings represent the only report of a chelonian egg wider than its pelvic canal, several other chelonian taxa have eggs wider than their AGs. This feature is prominent in the small endemics (Homopus spp., Psammobates spp., and C. angulata; M.D. Hofmeyr and B.T. Henen, unpublished data), Gopherus berlandieri (Agassiz, 1857) (Rose and Judd 1991) and Sternotherus odoratus (Latreille in Sonnini and Latreille, 1801) (Clark et al. 2001). It appears that a small AG has a strong selective value in these species. Sexual differences between conspecific males and females may indicate different selection pressures due to different needs of the two sexes. Relative to body size, the AG of female H. signatus was smaller than the AG of conspecific males. A small AG may confer an antipredator benefit, but females have to overcome this constraint when laying eggs. The large tail of males probably requires a large shell opening, particularly to manoeuvre the tail and penis during copulation. Testudo horsfieldii Gray, 1844 males also have larger posterior shell openings relative to the openings of conspecific females. Bonnet et al. (2001) postulated that T. horsfieldii males need large posterior shell openings to facilitate tail movement during copulation, and that large hind limb openings improve male mobility. Pelvic apertures in female H. signatus were larger than pelvic apertures in conspecific males. Similar differences were reported for G. berlandieri, Kinosternon flavescens (Agassiz, 1857), and Terrapene ornata (Agassiz, 1857) (Long and Rose 1989), indicating that natural selection favours larger pelvic apertures in females of these species. The pelvic girdle serves a locomotory function in H. signatus males and females. However, female pelvic girdles must accommodate a large egg at oviposition, necessitating a wider pelvic aperture than that required by males. Lateral views of H. signatus males and females indicate shape differences between sexes, with females having a higher carapace at the pelvic region than males. In this dorsoventrally flattened species, female shape allows more space for a large egg. Additionally, the ilia of H. signatus females tilt more to the posterior than do male ilia. This inclination of female ilia may facilitate passing the large egg by directing the long axis of the egg towards the AG on the posterior ventral surface. Shell and pelvic kinesis Shell kinesis in posterior elements of the female plastron and carapace probably facilitates laying large eggs (Rose and Judd 1991; Pritchard 1993). In species with shell kinesis developing ontogenetically, fibrous connections (syndesmoses) replace bony sutures of the kinetic bony elements (Pritchard 1993). In H. signatus, plastral and carapacial kinesis occur at oviposition, with plastral depression occurring posteriorly (xiphiplastron and hypoplastron) in concert with elevation of the pygal and peripheral bones of the carapace (Greig 1976; V.J.T. Loehr, unpublished data). Separation of the posterior scutes (supracaudal and posterior marginal, vertebral and pleural scutes), accompanied with stretching of the associated connective tissue, can be seen during the lifting of the carapace at oviposition (Greig 1976; V.J.T. Loehr, unpublished data). The severe kinesis emphasizes the selective advantage of producing such large eggs. Phylogeny may significantly influence the ability of chelonian pelves to accommodate large eggs. In pleurodiran chelonians, but not in cryptodires, the pelvis is fused (sutured) to the carapace and plastron (Ernst and Barbour 1989; Pough et al. 1999). In H. signatus, a cryptodire, the paucity of intact pelves in dry specimens supports the idea of syndesmotic connections for pelvic connections to the shell and at the pubic symphysis. Similar to mammalian parturition, the pelvis of H. signatus must exhibit extreme kinesis at oviposition, accommodating the egg and other tissues (e.g., rectum). This kinesis probably includes loosening of the pubic

1350 Can. J. Zool. Vol. 83, 2005 symphysis and freeing of the ilial heads from the sacral ribs and large suprasacral fossae. The mechanism that controls pelvic, plastral and carapacial kinesis during H. signatus oviposition is not known. In the marine turtle Caretta caretta (L., 1758) (Guillette et al. 1991), plasma estradiol appears to prepare prostaglandin receptors before nesting, and prostaglandins are involved in egg expulsion (uterine contractions and cervical relaxation). During mammalian parturition, relaxin loosens fibrous connections of the pelvis, especially the pubic symphysis, and softens the cervix (Koob 1998). Relaxin serves similar functions in avian oviposition and elasmobranch oviposition or parturition (Koob 1998). Hormones may coordinate shell and pelvic kinesis in H. signatus oviposition. Reproductive strategy and environmental effects Climatic conditions in southern Africa progressively aridified since the Oligocene (Partridge 1997) and a strong east-west aridity gradient started to develop in the late Miocene. Currently, mean AR ranges from more than 1000 mm year 1 in the east to less than 100 mm year 1 in the west, while rainfall is less variable in the east than in the northwest (CV of <20% in the east to >80% in the northwest; Lovegrove 1993; Schulze 1997). The distribution of multiple-egg and single-egg clutching taxa of the small endemics is consistent with the development of mesic and xeric conditions in southern Africa. It appears that aridity and unpredictable rainfall determined which taxa produce single-egg clutches. In arid and unpredictable environments, it can be prudent for females not to commit all of their resources to one clutch because females need to retain some reserves for their own survival. Within this scenario, tortoises producing single-egg clutches can respond to environmental conditions by adjusting clutch frequency. When resources are abundant, these tortoises can increase clutch frequency without compromising their reserves. By spreading their reproductive commitments over time, females may place eggs into different environmental conditions, increasing the likelihood that some eggs will hatch into favourable conditions (Congdon et al. 1982; Stearns 1992; Wallis et al. 1999). This strategy addresses the risk of producing too many offspring at one time, and the development of single-egg clutches among the small endemics is consistent with the bet-hedging concept. Although information is scarce, at least four of the single-egg clutching taxa can produce more than one clutch in a reproductive season (M. tornieri, Schmalz and Stein 1994; Homopus sp., Schleicher and Loehr 2001; H. signatus, Loehr et al. 2004; C. angulata, Hofmeyr 2004). There is probably a minimal size for viable eggs in arid and unpredictable environments. Among the small endemic taxa, EV increases where rainfall is less predictable, implying a selective advantage for large eggs in unpredictable habitats. The definition of what constitutes an optimal egg remains elusive (Congdon and Gibbons 1990), so egg size studies have been limited primarily to qualitative tests of predictions from optimal egg size theory. It appears that natural selection has not optimized egg size in H. signatus; egg size increased with maternal size and females in good body condition produced larger eggs than did females in poor condition. However, some evidence suggests that the largest H. signatus eggs approach an optimal size. First, EW and PW scaled less than proportionately to SCL. Additionally, EW scaled allometrically with PW and the regression line approaches the isometric line for PW. These factors, plus the small and similar coefficients of variation for EW and EL, may mean that some H. signatus females produce eggs that approach optimality. In H. signatus, heavy eggs produce heavy hatchlings (V.J.T. Loehr, unpublished data). The additional mass may be composed of somatic tissue as well as reserves for hatchling maintenance and growth (Congdon and Gibbons 1990). Large C. picta eggs have greater hatching success (Gutzke and Packard 1985), large turtle and tortoise hatchlings become large juveniles (Spotila et al. 1994; Janzen and Morjan 2002), and large Chelydra serpentina (L. 1758) hatchlings may have greater survivorship (Janzen 1993). Greater hatchling survivorship would improve maternal fitness, but large egg size does not always improve hatchling survival (Congdon et al. 1999). In the arid and unpredictable environment of H. signatus, a large hatchling size may be advantageous. Hatching may occur before enough rain falls to stimulate new plant growth during autumn or winter. In this event, hatchlings will require yolk or body reserves to survive unfavourable conditions. When food is available, larger hatchlings may acquire more water and other nutrient reserves that help them survive the subsequent hot, dry season, which can last 6 months or longer. The large eggs of H. signatus are central to a suite of extreme characters that confer selective advantages in a harsh and relatively unpredictable environment. Acknowledgements We are grateful to T.E.J. Leuteritz, F.A.C. Schmidt, A. Schleicher, and Western Cape Nature Conservation Board (E.H.W. Baard and A. de Villiers) for use of unpublished morphometric data. We thank K. le Roux of the Old Paarlweg Animal Hospital and staff of Dr. Van Niekerk hospital in Springbok for assistance in radiographing tortoises, and D. Hamerton for assistance with South African Museum specimens. We thank K. Nagy and G. Kuchling for useful comments on a draft manuscript. Field efforts were permitted by Northern Cape Nature Conservation (permit Nos. NNO 1/10/2 019/2001 and NNO 1/10/2 137/99). Research funding came from the National Research Foundation (South Africa), Royal Society (London), University of the Western Cape, Dutch Foundation for the Advancement of Herpetology, Tortoise Trust USA, Dutch Turtle/Tortoise Society, Seneca Park Zoo, and Basel Zoo. Research was completed according to the ASIH-SSAR-HL Guidelines for Use of Live Amphibians and Reptiles in Field Research and was authorized by the Research Ethics Committee, University of the Western Cape (permit 96/10/15). References Bonnet, X., Lagarde, F., Henen, B.T., Corbin, J., Nagy, K.A., Naulleau, G., et al. 2001. Sexual dimorphism in steppe tortoises (Testudo horsfieldii): influence of the environment and sexual selection on body shape and mobility. Biol. J. Linn. Soc. 72: 357 372.

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