Morphological, cytogenetic and allozymic variation withinCephalofovea(Onychophora: Peripatopsidae) with descriptions of three new species

Zoological Journal of the Linnean Society (1995), 114: 115–138. With 9 figures

Onychophora: past and present. Edited by M. H. Walker and D. B. Norman

Morphological, cytogenetic and allozymic variation within Cephalofovea (Onychophora: Peripatopsidae) with descriptions of three new species A. L. REID*, N. N. TAIT, D. A. BRISCOE Macquarie University, School of Biological Sciences, Sydney, N.S.W., Australia 2109 AND D. M. ROWELL Australian National University, Division of Botany and Zoology, Canberra A.C.T., Australia 2601

Variation within and between populations of Cephalofovea (Peripatopsidae) has been examined by allozyme, karyological and morphological analyses. Four groups are recognized on the basis of allozyme electrophoresis. One group includes specimens from the type locality of the only described species of the genus, C. tomahmontis. While karyotypic and morphological character states show considerable inter-group variation, the distributions of these states among groups are not concordant when different characters are compared. However, each group of populations is uniquely defined by the full suite of character states it possesses. The four groups are recognized here as distinct species with three species described as new. ADDITIONAL KEY WORDS:—Cephalofovea – sibling species – allozymes – chromosomes – taxonomy. CONTENTS Introduction . . . Material and methods . Results . . . . Allozymes . . Karyology . . Morphology . . Taxonomic conclusions Taxonomy . . . Incertae sedis . . Acknowledgements . References . . . Appendix . . . .

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* Correspondence to: Amanda Reid, Australian National University, Research School of Biological Sciences, Canberra A.C.T. 0200, Australia. 0024–4082/95/050115+24 $08.00/0

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© 1995 The Linnean Society of London

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A. L. REID ET AL. INTRODUCTION

The genus Cephalofovea Ruhberg et al., 1988 was erected for C. tomahmontis, a species of Onychophora found in the moist montane forests west of Sydney, New South Wales, Australia. The significant features of the original diagnosis of Cephalofovea include a characteristic dorsal head pit in both sexes and the presence of crural papillae on the underside of the first pair of oncopods in males. The head pit, a depression between and anterior to the eyes, corresponds to an eversible head structure in males. While some other Australian peripatopsids also possess pits (Tait & Briscoe, 1990), other features indicate that these belong to other, as yet undescribed genera. Since its description, more widespread sampling has extended the distribution of Cephalofovea some 150 km west of the type locality at Mt Tomah in the Blue Mountains to include sites through the Great Dividing Range and westward to the Canobolas Mountains. Specimens from localities within this range have been subjected to genetic and morphological analyses to determine the extent of intra- and inter-population variation and hence to establish whether all the populations are conspecific. MATERIAL AND METHODS

The study was based on material collected largely by the authors from 14 localities west of Sydney, New South Wales, Australia (Fig. 1, Table 1). Specimens include the type specimen and additional material, including paratypes from the type locality of C. tomahmontis. The number of specimens used for electrophoresis and karyology is shown in Table 1. Collection details of preserved material are included in the taxonomy section of this paper. Allozyme electrophoresis Specimens to be used for electrophoresis were decapitated and stored at −80°C prior to preparation of whole body tissue extracts. Protein extraction and electrophoresis methods are described in an accompanying paper (Briscoe & Tait, 1995: pp. 91–102). Most specimens were examined for 21 gene-enzyme systems. A reference homogenate derived from 30 Euperipatoides leuckartii (Saenger) from Mt Tomah was interspersed at regular intervals among the Cephalofovea samples across each gel. After measurement of all allozyme mobilities, the mobility of the most common allele (electromorph) found in the E. leuckartii sample was assigned a value of 1.000 and the Cephalofovea electromorph mobilities are expressed relative to this standard. Data analysis follows Briscoe & Tait (this volume), using the percentage fixed gene distance estimator of genetic distances (Richardson, Baverstock & Adams, 1986). Karyology Air dried chromosomal preparations were produced from testes as outlined in Rowell (1985) with the following modifications: (1) prior to fixation, testis tissue was treated for 10 min in a hypotonic solution consisting of insect saline diluted 2 : 1 with distilled water; (2) fixation was for 1–2 h. To ensure congruence of the data sets, the bodies of individuals used for karyotypic analysis were subsequently subjected to allozyme analysis.

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Figure 1. Collection sites of Cephalofovea specimens examined in this study. Lines indicate the 1000 m contour. Numbers refer to sites listed in Table 1.

Morphology Specimens were preserved following the method employed by Read (1988). Animals were anaesthetized by exposure to ethyl acetate vapour for 10 min, dipped quickly in 70% ethanol to render the cuticle less hydrophobic and soaked for approximately 1 h in cooled boiled water. Specimens were then fixed for 2 days in 4% formalin and stored in 70% ethanol. This standardized procedure resulted in extended, uncoiled specimens which could be easily measured and compared. At least two specimens of each sex from wellrepresented sites were dissected for examination of internal anatomy. Remaining body parts were utilized for scanning electron microscopy.

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TABLE 1. Collection sites of Cephalofovea and number of specimens used for allozyme electrophoresis and karyology ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — Nearest No. of specimens Site Latitude Longitude named locality +Allozymes Karyology 1. 33°29?S 150°28?E Mt Irvine 0 0 2. 33°44?S 150°27?E Hazelbrook 0 0 3. 33°33?S 150°25?E Mt Tomah* 11–24 3 4. 33°30?S 150°23?E Mt Wilson 3–7 1 5. 33°38?S 150°17?E Blackheath 0 0 6. 33°33?S 150°17?E Bell 2 0 7. 33°29?S 150°14?E Clarence 4–5 0 8. 33°29?S 150°02?E Rydal 6 1 9. 33°54?S 150°02?E Budthingeroo 1–3 0 10. 33°50?S 150°00?E Kanangra Boyd Nat. Pk. 6–8 1 11. ½33°23?S 149°55?S Sunny Corner 0 0 12. 33°58?S 149°48?E Black Springs 3 1 13. 33°14?S 149°47?E Mt Horrible 2 1 14. 33°21?S 148°59?E Mt Canobolas 4 1 ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — Nat. Pk.  National Park. *  Type locality of C. tomahmontis. + The lower number refers to the minimum number of specimens examined for all 21 gene-enzyme systems.

Scanning electron microscopy Specimens were dehydrated in an acetone series; 70%, 80%, 90% and 3×100% for 30 min each, critical point dried, gold coated and examined in a JSM 840 ( Japan Electron Optics Ltd. Japan) scanning electron microscope operated at 6 KV. Morphometrics and statistical methods The width of the head of each specimen, measured dorsally between the midpoints of each eye (HWE), was used as a size standard. This character was chosen as it was found to be highly correlated with total body length and appears to be less prone to variation depending on the degree of contraction of specimens. Eye diameter and width of each of the spinous pads on the third right oncopod were measured for each specimen, with pads numbered 1–3 from distal to proximal. Indices were calculated by dividing each measure by head width. Morphometric characters are expressed in descriptions as minimum-mean-maximum Statistical analyses were performed using ‘Statistix’ Version 2 (NH Analytical Software). To eliminate the effect of size variation between specimens, residual variables were calculated following the methods of Reid (1991), using HWE as a standard. For each comparative analysis, the regression was derived from the pooled set of data from all localities under consideration. Males and females were treated separately. Equations for the regression lines used to calculate residuals are given in the Appendix. Simple univariate analysis of variance (ANOVA) was used to test the significance of mean differences in residual variables (reflecting differences in sizes of individual body parts) among groups identified by electrophoresis. Variance between sample means was tested using an F test. A Bartlett’s test was used to examine the equality of within group variances. The null

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hypothesis in all analyses was that there was no difference in the values for a variable between groups. Character description Terms for morphology follow Ruhberg (1985), except that oncopod has been used in preference to leg to describe the distinctive onychophoran walking limb. Terminology used to describe the dermal papillae has been adapted from Read (1988). The term primary papillae refers to the large papillae aligned along the centre of the plical fold and bearing sensory spines. The secondary papillae are defined here as also aligned along the centre of the fold, but lacking sensory spines. Accessory papillae are much smaller and usually distributed along the edges of the plical folds. Plicae number and papillae counts were compared at equivalent positions on each specimen. Plicae number was counted between oncopods three and four and papillae were counted on plicae in line with oncopod number ten. Representative individuals from each population were photographed for examination for variation in colour and pigmentation pattern. Abbreviations used in text AM  Australian Museum, Sydney, NSW, Australia; ANIC  Australian National Insect Collection C.S.I.R.O. Division of Entomology, Canberra, ACT, Australia; EDI  eye diameter index; F  females; HWE  head width; J  juvenile; M  male; SP1I  spinous pad 1 index; SP2I  spinous pad 2 index; SP3I  spinous pad 3 index. RESULTS

Allozymes Differences in electrophoretic mobility among samples are interpreted as reflecting alternative mendelian alleles at the structural gene loci encoding the enzymes. In presumed heterozygotes, the number and intensity of bands observed were always consistent with the known secondary structure of the enzymes in eukaryotes (two bands for monomers, three bands for dimers, etc.), supporting this presumption (Richardson et al., 1986). Of the 21 loci screened, only three Idh c, Mdh c and Pk) were monomorphic across all samples, including the E. leuckartii standard. A further six loci (Aat A, Aat c, Fdp, Gapd, Mdh A, Pgam) were monomorphic within Cephalofovea but differed from E. leuckartii. For the remaining 12 potentially informative loci, alleles were shared between E. leuckartii and Cephalofovea populations for Acon (populations 3, 4, 6), Aldol populations 7, 8, 9, 10, 12, 13 and 14), Enol (populations 7, 8, 12, 13), Pgk (populations 3 and 4) an 6Pgd (population 6) (Table 2). Comparison of the allozyme profiles reveals that each Cephalofovea population differs from E. leuckartii at 16 or 17 of the 21 loci (76–81%) supporting the separation of these taxa at the generic level. Within each Cephalofovea population, average observed heterozygosity is relatively low for an invertebrate (cf. Selander, 1976), ranging from 0.014 to 0.161 with an overall mean of 0.078. In addition, there is considerable variation among populations (Table 2), with some alleles shared among geographically adjacent

2.7.4.3

4.1.2.13 4.2.1.11

1.1.1.8

2.7.1.1 1.1.1.42

5.3.1.8

1.1.1.44

5.3.1.9

2.7.2.3

2.7.5.1

Ak

Aldol Enol

aGpd

Hk IdhA

Mpi

6Pgd

Pgi

Pgk

Pgm

(30) (39) (1) (39) (1) (22) (38)

1.177 1.281 0.803 1.000 1.108 0.877

(2) (20) (4) (1) (17) (48)

1.409 (30)

0.947 (36)

1.221 1.723 1.818 0.769 1.079 0.560 0.872

0.731 (12) 1.000 (6) 0.708 (40) (6) (7) (1) (6)

0.877 (10)

1.000 (3) 1.108 (3)

1.281 (6)

1.409 (14)

0.947 (6)

0.560 (6) 0.872 (8)

1.221 1.723 1.818 0.769

0.731 (4) 1.000 (2) 0.708 (14) (1) (3) (4) (4)

0.877 (4)

1.108 (4)

1.281 (4)

1.000 (3) 1.409 (1)

0.947 (4)

0.560 (4) 0.872 (4)

0.769 (4)

0.500 0.708 1.221 1.723

1.000 (4)

(1) (15) (1) (14) (1) (12) (4) (12)

(15) (1) (16) (16)

0.877 (16)

0.894 0.947 1.594 1.980 2.472 1.177 1.281 0.759

0.769 1.079 0.750 0.872

1.000 (16) 1.723 (16)

0.708 (16)

0.731 (16)

(2) (4) (1) (5)

0.877 (6)

1.177 (2) 1.281 (4) 0.759 (4)

0.894 0.947 1.715 1.980

0.750 (2) 0.872 (6)

0.769 (6)

1.000 (4) 1.723 (2)

0.526 (1) 0.731 (1) 0.708 (4)

(8) (5) (3) (5) (3) (8)

0.877 (8)

0.803 (4) 0.889 (4)

1.177 (8)

0.560 0.752 0.872 0.947 1.070 1.594

0.769 (8)

1.000 (8) 1.723 (8)

0.611 (8)

0.731 (8)

(12) (1) (11) (12)

0.711 (12)

0.803 (8) 0.889 (4)

1.477 (12)

1.594 (12)

0.560 0.752 0.872 0.947

0.769 (12)

1.000 (12) 1.000 (12)

0.708 (12)

0.526 (12)

0.711 (4)

1.281 (2) 1.477 (2) 0.803 (4)

1.078 (4)

0.947 (4)

0.560 (4) 0.872 (4)

0.769 (4)

1.000 (4) 1.000 (4)

0.708 (4)

0.526 (4)

0.877 (6)

0.803 (1) 0.889 (5)

1.477 (6)

1.594 (6)

0.947 (6)

0.560 (6) 0.872 (6)

0.769 (6)

1.000 (6) 1.000 (6)

0.708 (6)

0.526 (6)

(8) (2) (2) (6) 1.281 (5) 1.477 (5) 0.889 (8)

0.947 1.024 1.594 1.767

0.750 (10) 0.872 (10)

0.769 (10)

1.000 (10) 1.000 (10)

0.708 (10)

0.526 (8)

0.877 (6) 0.969 (4) Obs h 0.057 0.042 0.083 0.087 0.143 0.083 0.014 0.083 0.029 0.161 ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– —

4.2.1.3

Acon

––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — Mt Tomah Mt Wilson Bell Kanangra Budthingeroo Mt Canobolas Rydal Mt Horrible Black Springs Clarence Enzyme E.C. No. 3 4 6 10 9 14 8 13 12 7

TABLE 2. Distribution of allozyme mobility states for twelve enzymes among ten populations of Cephalofovea. Each state is expressed relative to the mobility of the reference sample, E. leuckartii from Mt Tomah. Population numbers are shown in italics. Brackets refer to the number of times each allele was observed. Obs h  mean observed heterozygosity

120 A. L. REID ET AL.

VARIATION WITHIN CEPHALOFOVEA (ONYCHOPHORA) 1.000

1.409

1.000

1.108

121 1.221

populations. Alleles 6Pgd , 6Pgd , Pgk , Pgk and Aldol are confined to populations 3, 4 and 6 on the eastern edge of the Blue Mountains, alleles Enol 1.000, Pgi 1.477 and Acon 0.526 are found only in populations 7, 8, 12 and 13 further west, while Pgm 0.711 is restricted to populations 8 and 13. Similarly, allele Hk 0.750 is present in populations 7, 9 and 10 with Pgk 0.759, 6Pgd 1.715, 6Pgd 1.980 and 6Pgd 2.472 only in populations 9 and 10. Ak 0.611 is represented only in the isolated Mt Canobolas population, 14. These distributions do not support the hypothesis that the ten Cephalofovea populations are part of a large, freely interbreeding population. Rather they indicate major restriction of gene flow among some of the study populations. Further, there is little evidence for clines of gene frequency among populations; one gene pool appears to be replaced by another over relatively short geographical distances. For example, there are no fixed gene differences between populations 3 and 6 which are ½15 km apart but six fixed differences between 6 and 7 which are only ½10 km apart. Similarly, there are only two fixed differences between populations 12 and 13, separated by ½85 km but five fixed differences between populations 10 and 12 which are ½20 km apart. To identify pattern in the data set, the allozyme data were reduced to a matrix of genetic distances for all pairwise comparisons using the percent fixed gene difference measure of Richardson et al. (1986). Our sample sizes are too small to allow estimates of allele frequency to be made with acceptable accuracy. We have therefore only considered whether alleles are shared between populations. This approach yields a conservative estimate of genetic distance in the matrix, disregarding differences in frequency among populations. The matrix was then subjected to a complete linkage cluster analysis (Sneath & Sokal, 1973) to generate a dendrogram of putative relationships (Fig. 2). Four discrete groups of populations are apparent. Group I contains populations 3 (the type locality of C. tomahmontis), 4 and 6. Group II includes populations 9 and 10, Group III contains the population from Mt Canobolas, site 14, and Group IV contains populations 7, 8, 12 and 13.

Karyology Preliminary chromosome analysis lends support to the hypothesis that Cephalofovea populations represent distinct groups (Table 3). There is a major dimorphism among groups, with Group I and Group III populations sharing a chromosome number 2n  34, and Groups II and IV with chromosome number 2n  28. In some preparations, there is a distinct XY sex chromosome heteromorphism. Unfortunately, low levels of colchicine are not effective in producing fully-condensed metaphase spreads and higher levels are cytotoxic. Further, chromosome condensation is poorly coordinated between homologues. Accepting this limitation on our resolution, we cannot currently discern any further major chromosomal differences within either the 2n  34 or 2n  28 forms, nor determine whether the presence of a heteromorphic XY pair is characteristic of all Cephalofovea males.

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Figure 2. Dendrogram based on complete linkage clustering of percent fixed gene difference among ten Cephalofovea populations.

Morphology Variation in the distribution of crural papillae, shape of male head structure papillae, colour pattern and morphometrics was observed. Crural papillae The number of pairs of oncopods bearing crural papillae on male specimens varies among groups (Table 4). However, all specimens have crural papillae TABLE 3. Cephalofovea chromosome numbers for specimens collected from seven populations. Brackets refer to site numbers shown in Figure 1. *  none distinguishable ––––––––––––––––––––––––––––––––––––––––––––––––––– — Sex Group Population 2n determination I

Mt Tomah (3) 34 XY Mt Wilson (4) 34 * II Kanangra (10) 28 * III Mt Canobolas (14) 34 * IV Rydal (8) 28 * Black Springs (12) 28 * Mt Horrible (13) 28 XY ––––––––––––––––––––––––––––––––––––––––––––––––––– —

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TABLE 4. Crural papillae arrangement in male Cephalofovea. Number of specimens by locality with crural papillae on oncopod(s) 1, 1 & 2 or 1, 2 & 3. Brackets refer to site numbers shown in Figure 1 — –––––––––––––––––––––––––––––––––––––––––––––––––––––––––– Oncopod(s) 1 1 & 2 Group Population only only 1, 2 & 3 I

Mt Tomah (3) 11 1 Mt Wilson (4) 3 II Kanangra (10) 5 5 III Mt Canobolas (14) 1 7 IV Clarence (7) 7 1 Rydal (8) 15 3 Black Springs (12) 1 Mt Horrible (13) 3 — ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––

on the first pair of oncopods and none have crural papillae posterior to the third oncopod pair. In Groups I and IV, the majority of specimens have papillae only on the first pair of oncopods. In contrast, males of groups II and III always display crural papillae on the first two pairs of oncopods and frequently the first three pairs. These distributions are not size related and therefore, presumably independent of age and sexual maturity. Male head structure papillae Visible only when the head structure is everted, the shapes of papillae on the distal half of the structure of Groups I, II, and IV are essentially similar. They are dome-shaped (Fig 3A–D) and their bases display varying degrees of folding, which increases in larger, presumably older, specimens (compare A and C). The equivalent papillae seen in Group III (Mt Canobolas) specimens of varying sizes are flattened with no basal folding (Fig. 3E, F). Colour pattern Body colour and pattern are highly variable within Cephalofovea populations. In each population, two colour morphs, reflecting the dominant colour or hue, are found, one tan and the other greyish blue-violet. In Groups I, II and IV, in both colour morphs, there is a series of segmentally arranged Y-shaped mid-dorsal patches (Fig. 4A–E, G). These have been described in detail in Ruhberg et al. (1988). This pattern can be variably obscured due to the predominance of particular pigments with respect to others, seen in most extreme form in Figure 4D, E, G. The patterns shown in Figure 4D, E are found only on specimens from Mt Tomah (Group I, population 3) and Kanangra (Group II, population 10) and the pattern shown in Figure 4G occurs in low frequency in Clarence specimens (Group IV, population 7). The Y-shaped pattern is notably absent in specimens from Mt Canobolas (Group III). Figure 4F shows a blue-grey morph and Figure 4H shows a tan morph from this site. Other features of the colour pattern not given in Ruhberg et al. (1988) should be noted. Dorsal banding on the antennae rings is variable. Antennae rings are either (1) greyish blue and not banded or (2) grey and tan or tan

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Figure 3. A, Distal half of everted head structure C. tomahmontis male, HWE 1.62 mm (AM KS32325); B, Higher magnification of papillae, same specimen; C, Everted head structure C. cameroni sp. nov. male, HWE 1.68 mm (AM KS32341); D, Higher magnification of papillae, same specimen; E, Partially everted head structure, C. pavimenta sp. nov. male, HWE 1.35 mm (AM KS32351); F, Higher magnification of papillae, same specimen; G, Lateral head structure papilla C. cameroni sp. nov. male, specimen as in C; H, Lateral head structure papilla C. pavimenta sp. nov. male, specimen as for E. Scale bars, A–F  100 mm, G & H  10 mm.

VARIATION WITHIN CEPHALOFOVEA (ONYCHOPHORA)

Figure 4. Body pattern in Cephalofovea. A, C. clandestina sp. nov. female, HWE 1.66 mm (AM KS32337); B, C. tomahmontis female, HWE 1.39 mm (AM KS32324); C, C. tomahmontis female, HWE 1.22 mm (AM KS32324); D, C. clandestina sp. nov. female, HWE 1.48 mm (AM KS32335); E, C. clandestina sp. nov. female, HWE 1.48 mm (AM KS32334); F, C. pavimenta sp. nov. female, HWE 1.40 mm (AM KS32351); G, C. cameroni sp. nov. female, HWE 1.35 mm (AM KS32769); H, C. pavimenta sp. nov. Allotype, female, HWE 1.35 mm (AM KS32351). Scale bars1 mm.

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Figure 5. A, Enlarged papillae lateral to head pit C. cameroni sp. nov. male, HWE 1.51 mm (AM KS32341); B, Crural papilla oncopod 1 C. cameroni sp. nov. male HWE 1.68 mm (AM KS32341); C, Posterior ventral body C. pavimenta sp. nov. male HWE 0.90 mm (unreg); D, Posterior ventral body C. pavimenta sp. nov. female HWE 1.05 mm (unreg); E, Posterior accessory gland opening C. tomahmontis male HWE 1.62 mm (unreg). Scale bars A & E  10 mm; B, C & D  100 mm.

mottle rings alternate or (3) proximal half of each antennal ring tan banded or mottled. In (2) and (3) banding continues from the base approximately to the distal third of the antennae. Banding or mottle on the first three antennae rings and between the second and third ring often extends ventrally. In both sexes there is a pair of large papillae on either side of the head pit (Fig. 5A). These papillae in both sexes are usually tan and sometimes surrounded by a patch of tan papillae and:or tan integument. No females examined showed the pattern illustrated in Figure 3 of Ruhberg et al. (1988) of ten orange papillae framing the head pit. Papillae surrounding the anus are usually, but not always, tan. Morphometrics Bartlett’s test results for all pairwise analyses indicate that within group variances were not significantly different, justifying the comparison between

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groups using ANOVA. Analysis of variance between groups for the residuals from pooled regression lines revealed some significant morphometric differences (Table 5). In addition, for females, significant differences in size were found between Groups I (mean HWE 1.34) and IV (mean HWE 1.24) and between Groups II (mean HWE 1.39) and IV. TAXONOMIC CONCLUSIONS

The major features characterizing the four groups are summarized in Table 6. Each can be distinguished by a set of unique alleles and a varying combination of other characters. Group I includes all specimens form site 3, the type locality of C. tomahmontis, so specimens from sites 4 and 6 will also be referable to that name. The chromosomal differences between Groups II and IV versus Groups I and III can reasonably be assumed to form a barrier to gene flow, separating the former, geographically intermediate populations from the latter at the easternmost (Group I) and westernmost (Group III) extremes of the currently known distribution. Group III specimens are clearly morphologically distinct from C. tomahmontis (Group I). Although Groups II and IV do not differ markedly form C. tomahmontis (Group I) or from each other in qualitative morphological characters, they differ sufficiently electrophoretically to warrant formal taxonomic recognition. We therefore recognize four sibling species within Cephalofovea, three of which are described as new. The currently known distribution for these species is shown in Figure 9. TAXONOMY

Genus Cephalofovea Ruhberg et al., emended Cephalofovea Ruhberg et al. 1988: 120. Type species. Cephalofovea tomahmontis Ruhberg et al., 1988: 117–133, by monotypy. Diagnosis. Peripatopsidae with males possessing an eversible head structure consisting of a dome-shaped, fleshy crown. Distal half of crown with large, variously shaped, unpigmented papillae. Proximal part of crown unpigmented with scattered, pigmented papillae. Crural papillae present on first, first two or first three pairs of oncopods. Crural papillae absent on oncopods 4–15. Description. Antennae extending back to second oncopod pair, with 30 rings of uniform width or, in small specimens, alternating slightly narrower rings. Each antennal ring with single row of sensory spines. Distal 8–9 rings with sensory bulbs. Ventral surface of rings 4 or 5 to 13–16 (variable), counted for base of antennae, expanded to form pads bearing 2–3 rows of sensory spines. Males with eversible head structure consisting of fleshy dome shaped crown projecting anteriorly between antennae (Fig. 3). Distal half of everted crown with large variously shaped papillae with or without sensory spines. Papillae within this region with ribbed scales basally, and sharply demarcated whitish distal portion bearing fused or partially fused, blunt scales without ribs.

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TABLE 5. Cephalofovea males and females. Means and significance (Sig.) for analysis of variance between groups for residuals from pooled regression lines. n  sample size. ***P ³ 0.001, **P ³ 0.01, *P ³ 0.05, N:S  not significant ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — Variable n Group means Sig. (a) Males Groups I & II SP1 SP2 SP3 ED Groups I and III SP1 SP2 SP3 ED Groups I & IV SP1 SP2 SP3 ED Groups II & III SP1 SP2 SP3 ED Groups II & IV SP1 SP2 SP3 ED Groups III & IV SP1 SP2 SP3 ED

12 12 12 12 14 14 14 14 30 30 30 30 10 10 10 10 26 26 26 26 28 28 28 28

×Group I −0.005 −0.008 −0.004 0.001 ×Group I −0.002 −0.010 −0.001 0.003 ×Group I −0.004 −0.021 −0.002 0.002 ×Group II 0.007 0.002 0.009 0.005 ×Group II 0.004 −0.005 0.012 0.000 ×Group III −0.002 −0.005 0.000 −0.005

×Group II 0.010 0.015 0.007 −0.002 ×Group III 0.003 0.013 0.001 −0.004 ×Group IV 0.001 0.007 0.008 0.000 ×Group III −0.005 −0.001 −0.006 −0.003 ×Group IV 0.000 0.000 −0.002 0.000 ×Group IV 0.000 0.001 0.000 0.001

* * ** N:S N:S ** N:S N:S N:S *** N:S N:S N:S N:S * N:S N:S N:S ** N:S N:S N:S N:S N:S

(b) Females Groups I & II ×Group I ×Group II SP1 64 −0.002 0.002 N:S SP2 64 −0.012 0.012 *** SP3 64 −0.009 0.003 *** ED 64 0.005 −0.005 ** Groups I & III ×Group I ×Group III SP1 48 −0.005 0.010 *** SP2 48 −0.009 0.019 *** SP3 48 −0.005 0.010 ** ED 48 0.003 −0.005 N:S Groups I & IV ×Group I ×Group IV SP1 64 0.003 −0.003 * SP2 64 −0.005 0.005 N:S SP3 64 −0.001 0.001 N:S ED 64 0.000 0.000 N:S Groups II & III ×Group II ×Group III SP1 48 −0.004 0.007 * SP2 48 −0.003 0.005 N:S SP3 48 0.001 −0.001 N:S ED 48 −0.001 0.001 N:S Groups II & IV ×Group II ×Group IV SP1 64 0.005 −0.005 * SP2 64 0.004 0.004 N:S SP3 64 0.006 −0.006 ** ED 64 −0.004 0.004 * Groups III & IV ×Group III ×Group IV SP1 48 0.013 −0.006 *** SP2 48 0.010 −0.005 ** SP3 48 0.006 −0.003 * ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– —

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TABLE 6. Summary of diagnostic features of Cephalofovea spp. ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — Crural papillae Distal head Unique on Chromosomes structure Species Group alleles oncopods (2n) papillae Pattern C. tomahmontis

I

6Pgd 1.000, 6Pgd 1.409, Pgk 1.000, Pgk 1.108, Aldol 1.221

1 or 1 & 2

34

rounded

Y-shaped

C. clandestina sp. nov.

II

6Pgd 1.980, 6Pgd 1.715, 6Pgd 2.472, Pgk 0.759

1 & 2 or 1, 2 & 3

28

rounded

Y-shaped

C. pavimenta sp. nov.

III

Ak 0.611

1 & 2 or 1, 2 & 3

34

flattened

Y-absent

Enol 1.000, Acon 0.526, 1 or 1 & 2 28 rounded Y-shaped Pgi 1.477 ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– — C. cameroni sp. nov.

IV

Lateral and ventral papillae smaller with darkly pigmented, pointed apices. Proximal half of head structure with scattered pigmented papillae. Ventrally, proximal margin of crown with large, darkly pigmented papillae. When inverted, base of structure surrounding a depression or pit. Females with shallow depression between and anterior to eyes. Papillae within this region greatly reduced, crowded together. Both sexes with pair of enlarged, sometimes fused papillae on either side of depression (Fig. 5A). Inner jaw with 4–6 denticles, outer jaw with or without single small accessory tooth (Fig. 6A). Tongue with longitudinal row of 5 teeth (Fig. 6B), posterior 3 slightly larger than anterior pair. Large specimens often with a patch of 2–4 small teeth toward posterior end of median fold on either side of longitudinal row. Dorsum with 12 complete plicae and 2–5 anastomoses between oncopods 3 and 4. Wide and slightly narrower plical folds alternate. Number of papillae counted transversely from mid-dorsal line to junction of oncopod 10; males 10–14–20, females 9–19–32. Dorsal papillar arrangement: wider alternate folds with single primary papilla with short, narrow spine between a pair of primary papillae with longer, robust spines (Figs 6C & 7). At least one secondary papilla and various numbers of accessory papillae between primary papillae. Narrower folds with smaller, mainly secondary and accessory, papillae. Lateral primary papillae on both wide and narrow folds more elongate than those on rest of body. Additionally, one pair of more markedly elongate primary papillae between oncopods in line with junction of oncopods and body. Remaining integument with low smooth scales (Fig. 7A). Fifteen pairs of oncopods. Average length of last pair of oncopods approximately 75% length of preceding oncopod pair. Basal foot papillae absent. Spines on first and second spinous pads fine, evenly spaced, spines on third pad more widely spaced. A fourth broken spinous pad present. Nephridiopores with heavily ribbed scales basally, smooth margin surrounds a broad U-shaped opening. Body length posterior to genital opening approximately equal to length of last pair of oncopods. Papillae surrounding anal opening equal in size dorsally

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Figure 6. A, Outer (top) and inner (bottom) jaws C. tomahmontis female Allotype HWE 1.37 mm (ANIC). B, Mouth C. pavimenta sp. nov. Holotype, male HWE 1.58 mm (AM KS32338), lrt  longitudinal row of teeth on tongue; C, Diagram of papillae arrangement in Cephalofovea. Dots indicate sensory spines, large dots indicate robust spines. Median fine dot indicates short, delicate spine.

and ventrally. In both sexes, papillae posterior to genital opening large, conical (Fig. 5C, D). Females without ovipositor. Both male and female genital openings dimplelike, or approximately cruciform, not a distinct slit. Crural papillae and corresponding crural glands present on first pair of oncopods and variably present on second and third oncopod pairs. Papillae smooth and finely scaled basally and with a narrower, cylindrical portion distally, with broad, overlapping ribbed scales and smooth oval rim with longitudinal slitlike opening (Fig. 5B). Papillae protrude from 5th plicae counted from third spinous pad. Crural glands confined to oncopods, not enlarged within body cavity. Anterior accessory glands absent. Posterior accessory glands loosely coiled distally, open mid-ventrally, halfway between genital and anal openings (Fig. 5C, E).

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Figure 7. A, Integument C. tomahmontis female HWE 1.22 mm (AM KS32325). Large arrows indicate primary papillae with large robust spines. Small arrow indicates primary papilla with short delicate spine. Scale bar  100 mm; B, Primary papilla with large robust spine C. cameroni sp. nov. female HWE  1.31 mm (AM KS32341). Scale bar  10 mm; C, Primary papillae, delicate spine, specimen as in B. Scale bar 10  mm.

Male reproductive tract: vas deferens loops posteriorly immediately at junction of paired vasa efferentia Diameter of vas deferens approximately equal to diameter of proximal section of vasa efferentia (Fig. 8A). Female reproductive tract: ovarian walls thin, ova follicular, spermathecae and accessory organs present (Fig. 8B). Eggs heavily yolked. Embryos when present, found along entire length of paired uteri, each encased within a thin membrane. Embryos in uteri all appear to be at approximately at the same stage of development. Habitat. Rotting logs and leaf litter. Remarks. Specimens generally lie flat within rotting logs and are not prone to coiling when disturbed. The generic diagnosis has been modified to include characters found exclusively in Cephalofovea and no other genus. As a head pit is present in other undescribed genera, the diagnosis refers to the unique nature of the whole structure, readily determined when in the everted state. A number of specimens were found with spermatophores cupped within partially everted pits, as has been reported elsewhere (Ruhberg et al. 1988, Tait & Briscoe 1990). As crural papillae do not only occur on the first pair of oncopods, the diagnosis has been modified accordingly. Further characters may be added to the diagnosis when a full generic revision is completed. Ruhberg et al. mention that the uteri of C. tomahmontis do not unite at their distal region (Ruhberg et al., 1988, p. 127, Fig. 7). Approximately ten female specimens were dissected during the course of this study, including three from the type locality of C. tomahmontis. In all specimens, the paired uteri unite in the typical fashion to open via the gonopore. The accessory organs are not, as stated in Ruhberg et al. (1988), unique to Cephalofovea.

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Figure 8. A, Male reproductive tract. C. cameroni sp. nov. HWE 1.44 mm (AM KS32341), pa  posterior accessory gland, sv  seminal vesicle, t  testis, vd  vas deferens, ve  vas efferens. B, Female reproductive tract. C. cameroni sp. nov. HWE 1.26 mm (AM KS32341). ao  accessory organs, ctm  connective tissue muscle layer of ovary wall, e  egg, o  oviduct, st  spermatheca, u  uterus.

Cephalofovea tomahmontis Ruhberg et al., 1988. (Figures 3A, B; 4B, C; 5E; 6A; 7A; 9. Table 6.) Cephalofovea tomahmontis Ruhberg et al. 1988: 117–133, Figs 1–7. Material examined. AUSTRALIA: New South Wales – Mt Tomah 33°33?S 150°25?E Holotype 14.viii.1986, 1M (ANIC); Allotype 14.viii.1986, 1F (ANIC), Coll. N.N.T., V. Storch & families. Additional material. Mt Tomah 33°33?S 150°25?E, 1015 m, 15.v.1990, 3F (AM KS32324), Coll. N.N.T.; 23.viii.1991, 3M 6F (AM KS32325), Coll. N.N.T. & A.R.; 31.vii.1992, 5M 17F (AM KS32326), Coll. N.N.T. & Ray Cameron; Mt Wilson 33°30?S 150°23?E, 980 m, 23.i.1992, 4F 5J (AM KS32327), Coll. N.N.T. & Ray Cameron; 31.vii.1992, 2F (AM KS32328), Coll. N.N.T. & Ray Cameron. Diagnosis. With repeated segmental body pattern and distal half of male

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Figure 9. Distribution of Cephalofovea spp. identified in this study. , C. tomahmontis; Ž, C. clandestina sp. nov.; ž, C. cameroni sp. nov.; R, C. pavimenta sp. nov. Question marks indicate collection sites of indeterminate species.

head structure with rounded, scaled papillae mediodorsally. Chromosome number 2n  34, including a single X and Y chromosome. With a combination of genetic alleles shown in Table 2. The mobilities of the 6Pgd 1.000, 6Pgd 1.409, Pgk 1.000, Pgk 1.108 and Aldol 1.221 alleles are unique to this species. Measurements. (Type and additional material): HWE males 1.08–1.48–1.71 mm (Holotype 1.53 mm), females 0.99–1.34–1.58 mm (Allotype 1.37 mm); EDI males 0.05–0.06–0.08 mm, females 0.06–0.08–0.09 mm; SP1I males 0.04–0.05–0.06 mm, females 0.06–0.07–0.09 mm, SP2I males 0.06–0.07–0.09

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mm, females 0.08–0.11–0.14 mm; SP3I males 0.03–0.04–0.06 mm, females 0.03–0.05–0.07 mm. Remarks. Features distinguishing C. tomahmontis from C. clandestina sp. nov., C. cameroni sp. nov. and C. pavimenta sp. nov. are shown in Table 6. Cephalofovea clandestina sp. nov. (Figures 4A, D & E; 9. Table 6.) Material examined. AUSTRALIA: New South Wales – Kanangra Boyd National Park 33°50?S 150°00?E, 1140 m, Holotype 18.vii.1992, 1M (AM KS32329), Coll. N.N.T.; Allotype 18.vii.1992, 1F (AM KS32330), Coll. N.N.T.; Paratypes 6.ii.1990, 1F (AM KS32331), Coll. N.N.T. & A.R.; 8.v.1990, 1M 3F (AM KS32332), Coll. N.N.T.; 14.viii.1990, 5F 6J (AM KS32333), Coll. J. Miller; 22.viii.1991, 2F (AM KS32334), Coll. N.N.T.; 21.i.1992, 13F (AM KS32335), Coll. N.N.T. & Ray Cameron; 18.vii.1992, 2F (AM KS32336), Coll. N.N.T.; 14.viii.1992, 3M 4F (AM KS32337), Coll. N.N.T. Diagnosis. With repeated segmental body pattern and distal half of male head structure with rounded, scaled papillae mediodorsally. With 2n  28 chromosomes. With a combination of genetic alleles shown in Table 2. The mobilities of the 6Pgd 1.980, 6Pgd 1.715, 6Pgd 2.472 and Pgk 0.759 alleles are unique to this species. Measurements. (Type and additional material): HWE males 1.08–1.31–1.57 mm (Holotype 1.57 mm), females 1.08–1.39–1.66 mm (Allotype 1.66 mm); EDI males 0.05–0.06–0.07 mm, females 0.04–0.07–0.08 mm; SP1I males 0.06– 0.06–0.07 mm, females 0.05–0.07–0.10 mm; SP2I males 0.08–0.09–0.10 mm, females 0.10–0.13–0.15 mm; SP3I males 0.04–0.05–0.06 mm, females 0.04– 0.07–0.09 mm. Remarks. Features distinguishing C. clandestina sp. nov. from C. tomahmontis, C. cameroni sp. nov. and C. pavimenta sp. nov. are given in Table 6. Etymology. The specific name is derived from the Latin clandestinus meaning secret or hidden referring to this cryptic species.

Cephalofovea cameroni sp. nov. (Figures 3C, D, G; 4G; 5A, B; 7B, C; 8A, B; 9. Table 6.) Synonymy Cephalofovea tomahmontis Ruhberg et al., 1988; 120 (in part). Material examined. AUSTRALIA: New South Wales – Rydal 33°29?S 150°02?E, 900 m, Holotype 21.i.1992, 1M (AM KS32338), Coll. N.N.T. & Ray Cameron; Allotype 21.i.1992, 1F (AM KS32339), Coll. N.N.T. & Ray Cameron; Paratypes 16.ix.1989, 2M 1F (AM KS32340), Coll. N.N.T. & Ray Cameron; 21.i.1992, 11M 11F (AM KS32341), Coll. N.N.T. & Ray Cameron. Additional material. 6 km SSW of Mt Horrible 33°14?S 149°47?E, 1060 m, 23.ix.1987, 1F (AM KS32345), Coll. N.N.T.; 26.viii.1991, 1M 3F (AM KS32342), Coll. N.N.T.; 16.vii.1992, 2M 2F (AM KS32343), Coll. N.N.T.;

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Sunny Corner, 4.xii.1972 33°23?S 149°53?E, 1F (AM KS28192), Coll. J. Disney; x.1990, 1F (AM KS32346), Coll. G. S. Hunt; Rydal 33°29?S 150°02?E, 1.x.09, 1M 2F (AM KS32567, previously AMS K54138), Coll. Moreau; 1F (AM KS28202), Coll. Moreau; Clarence (near Lithgow) 33°29?S 150°14?E, 1080 m, 21.x.1992, 7M 6F (AM KS32769), Coll. A. R.; 1F (AM KS32352), Coll. Wells and Wellington; Lithgow 33°29?S 150°10?E, 17.iv.1972, 1F (AM KS28198), Coll. H. Disney & M. White; Black Springs 33°58?S 149°48?E, 1090 m, 22.i.1992, 6F 1J (AM KS32344), Coll. N.N.T. & Ray Cameron. Diagnosis. With segmentally repeated body pattern and distal half of male head structure with rounded, scaled papillae mediodorsally. With 2n  28 chromosomes. With a combination of genetic alleles shown in Table 2. The mobilities of the Enol 1.000, Acon 0.526 and Pgi 1.477 alleles are unique to this species. Measurements. (Type and additional material): HWE males 1.22–1.43–1.71 mm (Holotype 1.58 mm), females 0.90–1.24–1.62 mm (Allotype 1.31 mm); EDI males 0.04–0.06–0.07 mm, females 0.05–0.07–0.14 mm; SP1I males 0.03– 0.06–0.08 mm, females 0.05–0.07–0.09 mm; SP2I males 0.08–0.10–0.11 mm, females 0.07–0.12–0.15 mm; SP3I males 0.03–0.04–0.05 mm, females 0.04– 0.06–0.07 mm. Remarks. Features distinguishing C. cameroni sp. nov. from C. tomahmontis, C. clandestina sp. nov. and C. pavimenta sp. nov. are shown in Table 6. The two female and one male C. tomahmontis paratypes AMS K54138 (Ruhberg et al. 1988: 122, 2a) have been included with this species and given a new registration number, AM KS32567. Although indistinguishable morphologically from C. tomahmontis, specimens from Rydal clearly belong to a distinct species and it therefore seems likely that these preserved specimens, from the same locality as specimens examined electrophoretically in this study are referable to C. cameroni sp. nov. Etymology. This species has been named in honour of Mr Ray Cameron for his invaluable help in our study of Australian Onychophora which is greatly appreciated.

Cephalofovea pavimenta sp. nov. (Figures 3E, F, H; 4F, H; 5C, D; 6B; 9; Table 6) Material examined. AUSTRALIA: New South Wales – Mt Canobolas 33°21?S 148°59?E, 1395 m, Holotype 15.vii.1992, 1M (AM KS32347), Coll. N.N.T.; Allotype 15.vii.1992, 1F (AM KS32348), Coll. N.N.T.; Paratypes 6.ix.1989, 1M 2F (AM KS32349), Coll. N.N.T.; 27.viii.1991, 3F (AM KS32350), Coll. N.N.T.; 15.vii.1992, 6M 10F (AM KS32351), Coll. N.N.T. Diagnosis. Cephalofovea without repeated segmental colour pattern. Papillae comprising mediodorsal distal half of male head structure distally flattened, smooth without distinct scales. With 2n  34 chromosomes. With a combination of genetic alleles shown in Table 2. The mobility of the Ak 0.611 allele is unique to this species. Description. Measurements (type and additional material): HWE males 1.04–

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1.40–1.67 mm (Holotype 1.58 mm), females 1.08–1.30–1.53 mm (Allotype 1.35 mm); EDI males 0.04–0.05–0.07 mm, females 0.05–0.07–0.09 mm; SP1I males 0.03–0.06–0.08 mm, females 0.06–0.08–0.10 mm; SP2I males 0.08– 0.09–0.10 mm, females 0.11–0.13–0.15 mm; SP3I males 0.03–0.04–0.05 mm, females 0.04–0.06–0.10 mm. Colour and Pattern (preserved). Two colour morphs present both sexes. (1) Dark greyish-blue ground colour without pattern (Fig. 4F). Ventral pigment present, slightly paler than dorsum without pattern. Antennae not banded. Area between gonopore and anal opening pigmented as for rest of ventrum. (2) Tan ground colour with grey mottle dorsally, becoming darker dorsolaterally (Fig. 4H). Laterally, dorsal to oncopods a broad tan band without scalloped dorsal margin. With narrow dark grey band dorsal to an extending between oncopods. Ventrally pale tan with band grey mottle mid-ventrally extending to base of oncopods and along coxal furrows. Antennae rings dark bluish grey with traces of tan mottle dorsally. Ventrally with trace of tan mottle on antennae rings 2 (counting from base) and 3. Gonopore to anal opening tan medially, grey laterally. Gonopore papillae pale tan. Head structure. Males with eversible head structure consisting of a fleshy dome shaped crown which projects between antennae. Distal half of everted crown with large closely packed papillae (Fig. 3E, F). Mediodorsal papillae within this region scaled basally with regular margins (not folded) and smooth, flattened apices without scales. Papillae surrounding this region smaller, pointed, also with smooth apices without scales (Fig. 3H). Proximal half of head structure with scattered pigmented papillae dorsally. Laterally and ventrally, papillae with strongly hooked, darkly pigmented tips. Proximal ventral margin of crown with large dark greyish blue papillae. Remarks. Features distinguishing C. pavimenta sp. nov. from C. tomahmontis, C. clandestina sp. nov. and C. cameroni sp. nov. are shown in Table 6. The description of the colour pattern given in Ruhberg et al. (1988) is generally applicable to all species with the exception of C. pavimenta sp. nov. Only those features found to differ between the two C. pavimenta sp. nov. colour morphs or from those described in Ruhberg et al. 1988 are given here. Etymology. The specific name derived from the Latin pavimentum refers to the pavement-like appearance of the modified head structure papillae in males. Incertae sedis Cephalofovea sp. indet. Cephalofovea Ruhberg et al., 1988: 120. Material examined. AUSTRALIA: New South Wales – Mt Irvine ½33°29?S 150°28?E, 20.ix.1943, 5F (AM KS28211), Coll. Copland & Scrivaner; Hazelbrook ½33°44?S 150°27?E, 1F (AM KS28206), Coll. Abrahams; 1M 5F (AM KS32568); Blackheath ½33°38?S 150°17?E, 30.ix.1990, 1F (AM KS32353), Coll. M. le Breton; vi.07, 1M (AM KS28201), Coll. T.S. Remarks. Given the cryptic nature of species within this genus, until specimens from Blackheath, Hazelbrook and Mt Irvine have been compared

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electrophoretically, specimens from these localities are retained as doubtful species. C. tomahmontis type material listed in Ruhberg et al. includes 5 females and 1 male (AM KS32568) without locality data (Ruhberg et al. 1988: 122, 2b). Notes included with the specimens indicate that these specimens were used for figures 1, 2b, 2c, 5, 6 and 7 in Ruhberg et al. (1988). As additional species of Cephalofovea have now been recognized, the status of these paratypes is not known.

ACKNOWLEDGEMENTS

The authors wish to thank Dr Mike Gray (Australian Museum, Sydney) and Dr Bruce Halliday (CSIRO Division of Entomology, Canberra) for the loan of specimens, to Ray Cameron for his extended help with fieldwork and for technical assistance with electrophoresis and Dr George McKay (Macquarie University) for conducting the final analysis of the electrophoretic data, Angela Higgins (Australian National University) for technical assistance in the preparation of chromosome spreads, Jenny Norman and Annabell Gregor for printing photographs, Dr Hannelore Paxton for encouragement, advice and comments on the manuscript. A. Reid is supported by an Australian Postgraduate Research Award. The New South Wales National Parks and Wildlife Service and Forestry Commission issued us with permits to collect specimens in N.S.W. This is publication No. 156 of the Research Unit for Biodiversity and Bioresources.

REFERENCES Briscoe DA, Tait NN. 1995. Allozyme evidence for extensive and ancient radiations in Australian Onychophora. Zoological Journal of the Linnean Society 114: 91–102. Read VM St J. 1988. The application of scanning electron microscopy to the systematics of the neotropical Peripatidae (Onychophora). Zoological Journal of the Linnean Society 93: 187–223. Reid, A. 1991. Taxonomic review of the Australian Rossiinae (Cephalopoda: Sepiolidae), with a description of a new species, Neorossia leptodons, and redescription of N. caroli (Joubin 1902). Bulletin of Marine Science 49(3): 748–831. Richardson BJ, Baverstock PR, Adams M. 1986. Allozyme Electrophoresis. A handbook for Animal Systematics and Population Studies. Sydney: Academic Press. Rowell DM, 1985. Complex sex-linked fusion heterozygosity in the Australian huntsman spider Delena cancerides (Araneae: Sparassidae). Chromosoma 93: 169–176. ¨ kologie, Chorologie und Ruhberg H. 1985. Die Peripatopsidae (Onychophora). Systematik, O phylogenetische Aspekte. Zoologica 137: 1–183. Ruhberg H, Tait NN, Briscoe DA, Storch V. 1988. Cephalofovea tomahmontis n. gen; n. sp; an Australian peripatopsid (Onychophora) with a unique cephalic pit. Zoologischer Anzeiger 221: 117–133. Selander RK. 1976. Genetic variation in natural populations. In: Ayala FJ, ed. Molecular evolution. Massachusetts: Sinauer Associates, 21–45. Sneath PHA, Sokal RR. 1973. Numerical Taxonomy. The Principles and Practice of Numerical Classifications. San Francisco: WH Freeman. Tait NN, Briscoe DA. 1990. Sexual head structures in the Onychophora: unique modifications for sperm transfer. Journal of Natural History 24: 1517–1527.

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Regression equations for Cephalofovea males and females. Regression was used to calculate residuals (Y  a+bHWE) where Y  predicted value of that dependent variable, a intercept, b  slope, Sig.  significance of regression model: ***P ³ 0.001, **P ³ 0.01, *P ³ 0.05 and r2  variance proportion explained by the particular model. –—–––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– Dep. Males Females a b Sig. r2 a b Sig. variable r2 Groups I & II SP1 0.264 0.037 0.029 N:S 0.482 −0.008 0.008 *** SP2 0.275 0.043 0.051 N:S 0.524 −0.063 0.165 *** SP3 0.351 0.094 −0.024 * 0.223 −0.003 0.063 *** ED 0.521 0.008 0.055 ** 0.414 0.003 0.071 *** Groups I & III SP1 0.189 0.024 0.036 N:S 0.043 −0.022 0.093 *** SP2 0.238 0.055 0.044 N:S 0.250 0.025 0.097 *** SP3 0.009 0.055 0.000 N:S 0.165 0.010 0.050 *** ED 0.513 −0.004 0.061 ** 0.355 −0.006 0.079 *** Groups I & IV SP1 0.000 0.077 0.001 N:S 0.477 −0.007 0.076 *** SP2 0.059 0.091 0.027 N:S 0.393 0.013 0.102 *** SP3 0.033 0.043 0.010 N:S 0.312 0.008 0.048 *** ED 0.265 0.035 0.036 *** 0.417 −0.009 0.083 *** Groups II & III SP1 0.311 −0.002 0.006 N:S 0.377 −0.010 0.085 *** SP2 0.761 0.005 0.090 *** 0.735 −0.085 0.191 *** SP3 0.081 0.044 0.013 N:S 0.351 −0.002 0.084 *** ED 0.876 −0.070 0.108 *** 0.451 −0.004 0.073 *** Groups II & IV SP1 0.007 0.071 0.007 N:S 0.467 −0.014 0.082 *** SP2 0.377 0.045 0.064 *** 0.707 −0.073 0.178 *** SP3 0.100 0.031 0.020 N:S 0.453 −0.029 0.084 *** ED 0.486 −0.008 0.066 *** 0.393 0.003 0.070 *** Groups III & IV SP1 0.017 0.061 0.014 N:S 0.372 −0.028 0.095 *** SP2 0.284 0.070 0.047 *** 0.577 −0.039 0.153 *** SP3 0.381 0.005 0.037 *** 0.496 −0.034 0.087 *** ED 0.495 −0.016 0.070 *** 0.320 −0.009 0.081 *** ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– —