Search

Supplementary figures, text and tablesMahony_et_al_A_new_hip_pocket_frog_from_eastern_Australia_Supplementary FigS1 DFA morph plot 5June2021 both sexesMahony_et_al_A_new_hip_pocket_frog_from_eastern_Australia_Supplementary_FigS2_Assa darlingtoni_photos of most regional locations_no_overlap with Fig 8Mahony_et_al_A_new_hip_pocket_frog_from_eastern_Australia_Supplementary_FigS3_Assa_call_DF_regressed_against_temperatureMahony_et_al_A_new_hip_pocket_frog_SUPPLEMENTARY_TABLES_S1-6Mahony_et_al_A_new_hip_pocket_frog_SUPPLEMENTARY_TEXT_TABLE_S6_FIGURE_S3_effect of temperature on call traitsMahony_et_al_A-new_hip_pocket_frog_SUPPLEMENTARY TEXT_TABLE S7_Conservation Assessment

Assa wollumbin sp. nov. Mount Wollumbin hip-pocket frog Figs 12, 13. Holotype: AMS R185959 adult male, collected on the forest floor at Wollumbin (Mt Warning) National Park, 28 o 23��� 55��� S, 153 o 17��� 01��� E, north-eastern New South Wales, by Stephen Mahony, on 17 October 2014. Diagnosis. Assigned to Assa based on the generic definition of Tyler (1972). Assa wollumbin sp. nov. can be distinguished by apomorphic nucleotide states at 30 sites in the ND2 gene (Tables 2, 3). Measurements of holotype (mm). SVL 15.8; HL 4.7; HW 6.3; ED 1.9: EN 1.1; IND 2.2; FL 10.2, TL 6.6; HHL 4.3; FLL 2.6. Description of holotype (Fig. 12). AMS R185959. An adult male. Small (SVL 15.8 mm), body pear-shaped, flattened dorso-ventrally, with a prominent dorso-lateral margin demarcated by a strong contrast in colouration and the presence of a line of small tubercles that extends from behind the eye to the posterior third of the dorsum. The pouch opening is on the dorso-lateral line in the anterior groin but is usually hidden by the thigh when the animal is at rest (Fig. 14E���F). Brood pouch openings on both upper lateral posterior flanks. Head moderately wide (HW/HL = 1.4). Canthus rostralis slightly concave; nostrils closer to tip of snout than to eyes; internarial distance greater than distance from external naris to eye (IND/EN = 1.8); snout slightly pointed when viewed from above; downward wedge shaped when viewed laterally. Eyes moderately long (ED/HL = 0.4), prominent, not protruding in dorsal and ventral views; pupil horizontal and elliptical. Tympanum present but obscured by granular skin, and substantially smaller than the eye. Glandular ridge of enlarged tubercles from rear of jaw to axil. Skin of dorsal and lateral surfaces including limbs, finely granular. Ventral surfaces including limbs coarsely granular covered with small tubercles and lateral edges and margin of jaw with more prominent tubercles. Urostyle prominent and rounded and upturned above the cloaca. Posterior, hidden surface of the thighs with many moderately sized rounded tubercles and strongly contrasting in colour with the dorsal and ventral surfaces. Arms moderately long, thin, with a row of moderate sized tubercles on the posterior dorso-ventral margin of the forearm. Fingers short without webbing; finger relative lengths 3> 2> 4> 1; finger one greatly reduced, only one phalanx; tips of all fingers slightly expanded laterally and smooth and rounded below; palmar surfaces with a large round and flattened tubercle at the outer heel; subarticular tubercles on each phalanx of the fingers. Legs moderately long (TL/SV = 0.4), thigh robust and muscular. Row of prominent tubercles on the posterior dorso-lateral margin of the tarsus and heel. Foot relatively long (FL/SVL = 0.65). Toes unwebbed, relative lengths 4> 3> 5> 2> 1; tips of all toes simple slightly expanded laterally and smooth and rounded below; rounded subarticular tubercles at base of each phalanx; small rounded outer metatarsal tubercle. Variation. A small frog, females are slightly larger than males (SVL: males range 13.1���19.3 mm, females range 14.7���20.5 mm) (Table 5). Head moderately wide to wide (HW/HL: range 1.2���1.7). Eye size variable (ED/HL: range 0.3���0.5); internarial distance substantially greater than distance from eye to naris (IND/EN: range 1.6���2.5). Hind limbs moderately long (TL/SVL: range 0.4���0.5). Distinct dorso-lateral ridge formed by rounded tubercles that vary in size between individuals from small to prominent. Some individuals with one or two larger rounded tubercles coincident with the v-shaped markings on the dorsum. Tubercles along the posterior dorso-lateral margins of the tibia and tarsus, and larger tubercles at the elbow and heel, more prominent in some than others. Colour and pattern. Highly variable dorsal and ventral colouration and patterning, no obvious differences between the sexes (Figs 12, 13). Base colour of the dorsal surface of head, body, legs and arms highly variable from light cream with an orange tinge to reddish brown to a darker brown and brown-black (Figs 12, 13). In many individuals with a lighter colouration there are distinct darker brown to black markings, which in many individuals occur as two anterior direct v-shaped darker chevrons on the upper and lower dorsum. The lower marking is continuous with a transverse dark bar on the thigh and foot when folded next to the body (Figs 12, 13). In other individuals there is a broad darker mid-dorsal stripe with distinct broad lighter cream and black margins (Fig. 13F). The mid-dorsal stripe may be mottled. A faint interorbital bar that does not extend up onto the orbit in most individuals, and in those individuals with a mid-dorsal stripe it forms the anterior margin. The face may be the same colour as flanks to a darker hue and may be immaculate or with ill-defined black mottling. Canthus rostralis with or without a darker marking between the anterior margins of the eye to the nostril. A dark vertical bar beneath the eye to the maxilla in most specimens, may be obscured in darker individuals, and occasionally with a cream lined edge. In most individuals there is a distinct delineation between the dorsal surface and the flanks marked by a ridge that is accentuated in individuals where the colouration of the dorsum and flanks is highly contrasting. Often there is a thin lighter coloured line on the ventral side of the ridge. Colouration of the upper lateral surface is dark brown to black diffusing into a lighter brown towards the ventral surface. Ventral surfaces including groin, hind and forelimbs, hands and feet, posterior surface of thigh and chin are the same colour within an individual (Fig. 13B, G���I). However, the colour differs considerably among individuals, from darker brown-black with small lighter coloured granules that are more prominent on the edges of the mandibles, to light burnt-yellow with darker flecks. Iris black with a fine golden inner rim and fine golden flecks dorsally. Reproductive mode and tadpole description. The occurrence of the hip-pockets in the male is a distinctive form of sexual dimorphism, and the opening of the pouch can be identified readily in the field with the use of a hand lens. The lateral pouch opening in the inguinal region is up to 3 mm long, runs laterally and is inclined posteriorly at a slight angle on the upper dorso-lateral margin. The pouch is continuous with the dermis and extends forward into the lymph sac beneath the lateral and ventral surfaces. The right and left pouches are independent and do not join. There is no evidence of a nuptial pad on the dorsal surface of the fingers or prepollex in any of the male specimens that were collected while calling and reproductively active. The mating sequence has been observed on more than ten occasions during a study of male parental care in Assa conducted at Wollumbin (MJM pers. obs.). When calling, males expand the skin below the chin, however the vocal sac is not loose (see Fig. 14A). Males call from secluded positions usually beneath dense leaf litter, from curled-up leaves and in crevices among small rocks (Fig. 14B). Males establish a calling position and females are attracted to the male who does not move away from the calling position.After a female has been attracted to the calling position, the male rapidly moves to an inguinal embrace. After embrace the female moves away, with male attached, to the site of oviposition which is up to 20 cm from where the male called. This indicates that the male is not calling from a microhabitat that represents a preferred oviposition site. Oviposition occurs beneath leaf litter or in a similar hidden location that is moist but not in standing water. The small clutch of large eggs is placed closely, the eggs adhere to one another and there is usually at least two layers (Fig. 14C, D). Embryonic development proceeds for a period of at least seven days, and the developing embryo can be seen through the thick transparent jelly coat (Fig. 14D). The adult male takes up a position about two or three body lengths away from the clutch, also under the leaf litter and in a position where he can view the clutch (Fig. 14E). At times the male will move to the clutch and move over it. We can only speculate on the cues that the males rely on to indicate the time of embryo hatching. However, the embryos do wriggle vigorously at times and this may provide some cue for the male. At hatching the male moves over the clutch, and the jelly liquifies. The embryos wriggle vigorously, with the head arched downward. The male rotates slowly within the clutch, and the legs are held slightly away from the body so that a crevice is created between his flanks and legs. The flanks are slightly concave. Newly hatched tadpoles wriggle up the crevice to the opening of the pouch in the inguinal region (Fig. 14F). Apart from rotating in the liquified clutch the male does not assist tadpoles into the pouch. Our observations are that most carrying males (n=12) have approximately equal number of tadpoles in the left and right pouch. Males have a rounded and robust body habitus when carrying tadpoles, and the pouch opening is no more distinct than in non-carrying males. Metamorphosis occurs in the pouches and the young emerge, usually hind limbs first (Fig. 14F), without any prompting or assistance, such as body contractions by the male. Our observations of embryonic and tadpole development (n = 6) in A. wollumbin sp. nov. are very similar to those described in detail for A. darlingtoni across its range by Anstis (2013). However, we observed two aspects of reproductive behaviour not described previously. First, we found several cases in which a male carried two cohorts of tadpoles, and the difference in development stage was such that it can only be that the male had incorporated two clutches that were fertilised well separated in time. In these cases, the brooding male was actively calling when collected. Second, when a female approached a male to enter into amplexus we did not see any evidence that the female inspected the male prior to embrace as described by Ehmann & Swan (1985). The two observations are linked in that we observed no evidence that the female inspects the male to see if he is holding tadpoles in his pouches prior to mating, and as such this is direct evidence that a male carrying young will mate a second time in the one breeding season. Call. Male advertisement calls have a duration of about 1.2 s (mean 1.24 s; range 0.8��� 1.8 s) and comprise a series of fully modulated repeated notes of the same duration (mean 0.046 s; range 0.045���0.05) separated by an inter-note interval that is slightly longer (mean 0.046 s; range 0.045���0.05) (Fig. 6). Inter-note time is even across most of the call but is slightly longer prior to the last note. The number of notes ranges from 5 to 14 (Table 6). There is only very slight amplitude modulation in the call with the first few notes being lower in energy than the subsequent notes. The call is broadband with a relatively high dominant frequency (mean 3743, range 3593���3992 Hz), and the lower mean 5% and upper mean 95% frequencies (3093 and 4218 Hz respectively), with the lowest and highest frequencies from 2906 to 14514 Hz (Fig. 6). There is no evidence of frequency modulation across the call or within notes (Fig. 6). Temporal and structural attributes of the call are affected by temperature. Call duration, inter-call duration and NRR are significantly negatively correlated with temperature (Table 6), while the number of notes in the call is positively correlated with temperature (Supplementary Table S2). Structural and temporal attributes of the call can also be affected by behavioural context. Males synchronise calls in Assa darlingtoni and can add or delete notes from the call and alter the timing between calls and adjust the note repetition rate (Clulow et al. 2017).). We have observed synchronous calling in A. wollumbin sp. nov. The courtship call is made when a male is approached closely by a female, at amplexus and possibly at times when the males are in very close proximity (Etymology. Referring to the distribution, which is primarily on Wollumbin, a noun in apposition. Wollumbin is the name given by the First Nation Traditional Custodians for the Tweed, and the mountain that is the cone of the Tweed volcano (also known as Mount Warning). Distribution. Known only from Wollumbin (Mt Warning) in north-eastern NSW (Fig. 8). The discovery of two species of Assa, in close proximity in the Tweed Volcano region, with one on the central volcanic cone Wollumbin and the other on the surrounding caldera and its slopes, is unexpected. The distance between sampling locations on Wollumbin and those on the caldera on four side are between 12 and 17 km (Fig. 8). The caldera is about 32 km in diameter, and Wollumbin is in the centre, however there is significant relief within this small distance with Wollumbin (1068 m above sea level [asl]) and the higher parts of the caldera (1040 m asl), are separated by the caldera valley below 100 m asl. The natural barrier to gene flow between A. wollumbin sp. nov. on Wollumbin and A. darlingtoni on the caldera to the east, south, west and north are unknown. A preference for cool temperate forest at higher altitudes and avoidance of lower and hotter forests does not appear to be the likely barrier. While Assa are commonly found in the cooler temperate forests at higher altitude, they also occur at lower altitudes in subtropical rainforest in the Currumbin Valley (160 m asl), at Terania Creek, Nightcap Range (220 m asl) and on Wollumbin (420 m asl). Prior to European settlement and clearing of parts of the Tweed Valley, the natural vegetation was continuous between Wollumbin and the higher altitudes of the caldera rim (McDonald 2010, McDonald & Hunter 2010). Indeed, there is an almost continuous closed forest connection across this barrier in several locations at the present time, and this is especially the case along the small streams where the riparian zones have an almost complete rainforest understorey. Regardless of the removal of forest connections in the past 200 years, the level of genetic differentiation uncovered means that the break in gene flow between A. wollumbin sp. nov. and A. darlingtoni greatly pre-dates these events. It is possible that A. wollumbin sp. nov. and A. darlingtoni occur sympatrically in locations that have not been sampled and that a reproductive isolating mechanism prevents gene flow, or that the current geographic barriers are not what drove the speciation. We found no evidence of sympatry in the genetic screening between the species, however the extent of our sampling on Wollumbin and its surrounding foothills is limited (Fig. 8). We are uncertain about the species identity of Assa from three locations (Mount Nullum and Wollumbin State Forest) within the Tweed Caldera within close proximity of Wollumbin (Fig. 8), and identification of samples from these locations would provide a firmer understanding of the distribution of the two species within the caldera., Published as part of Mahony, Michael J., Hines, Harry B., Mahony, Stephen V., Moses, Bede, Catalano, Sarah R., Myers, Steven & Donnellan, Stephen C., 2021, A new hip-pocket frog from mid-eastern Australia (Anura: Myobatrachidae: Assa), pp. 451-486 in Zootaxa 5057 (4) on pages 474-479, DOI: 10.11646/zootaxa.5057.4.1, http://zenodo.org/record/5705530, {"references":["Tyler, M. J. (1972) A new genus for the Australian leptodactylid frog Crinia darlingtoni. Zoologische Mededelingen. Leiden, 47, 193 - 201.","Anstis, M. (2013) Tadpoles and frogs of Australia. New Holland Publishers, Sydney, 827 pp.","Ehmann, H. & Swan, G. (1985). Reproduction and development in the marsupial frog, Assa darlingtoni (Leptodactylidae, Anura). In: G. Grigg, R. Shine & H. Ehmann (Eds), Biology of Australasian frogs and reptiles. Surrey Beatty & Sons Pty Ltd, Chipping Norton and NSW in association with the Royal Zoological Society of New South Wales, New South Wales, pp. 279 - 285.","Clulow, S., Mahony, M. J, Elliott, L., Humfeld, S. & Gerhardt, H. C. (2017) Near-synchronous calling in the hip-pocket frog Assa darlingtoni. Bioacoustics, 26, 249 - 258. https: // doi. org / 10.1080 / 09524622.2016.1260054","McDonald, W. & Hunter, J. (2010) Part 2. The Natural Environment. Chapter 11. The Vegetation. In: Kitching, R, Braithwaite, R. & Cavanaugh, J. (Eds.), Remnants of Gondwana. A Natural and Social History of the Gondwana Rainforests of Australia. Surrey Beatty & Sons Pty Ltd, Chipping Norton, pp. 129 - 134."]}

The hip-pocket frog (Assa darlingtoni), a small terrestrial myobatrachid frog found in mid-eastern Australia, has a highly derived, unusual, reproductive mode involving a unique form of male parental care. Males have subcutaneous pouches that open near the hip, and the developing tadpoles are carried in these pouches to post metamorphosis. It is found on several isolated mountain ranges in closed forest habitats, associated with high rainfall and temperate or sub-tropical climates. We established genetic relationships among specimens sampled across the range using phylogenetic analyses of thousands of single nucleotide polymorphisms (SNPs) from the nuclear genome and mitochondrial ND2 gene nucleotide sequences. These analyses uncovered two lineages that are genetically distinct in both nDNA and mtDNA analyses and that have low levels of divergence in male advertisement calls and are morphologically cryptic. Our data support separate species status for each lineage, based on the molecular genetic data. The first, which we name as a new species, Assa wollumbin sp. nov., is restricted to a single mountain, Wollumbin (= Mount Warning), the eroded cone of an ancient shield volcano—the Tweed Volcano. The second, the nominal species A. darlingtoni, has a wider distribution in five geographically disjunct subpopulations along 430 km of the Great Dividing Range in south-eastern Queensland and north-eastern New South Wales. The distributions of the two species closely approach within 15 km of each other on the central plug and rim of the caldera of the Tweed Volcano. Assa wollumbin sp. nov. meets the conservation criteria for Critically Endangered [A3(e), B2(a,b)]. When all subpopulations of A. darlingtoni are combined the conservation assessment is Endangered [A3(e), B2(a,b)]. Because of the fragmented nature of the distribution of A. darlingtoni, combined with the genetic evidence of concordant sub-structuring, we also conducted a conservation assessment on the five subpopulations. Two were assessed as Critically Endangered (D’Aguilar Range and Conondale/Blackall Ranges), and the remainder as Endangered (Dorrigo Plateau, McPherson Ranges, and Gibraltar Ranges/Washpool).

Litoria spaldingi (Hosmer, 1964) Northern Creek Frog (Figs 6, 9, 10) Hyla spaldingi Hosmer, 1964 Holotype. An adult female, WAM R 23886 (formerly AMNH No. 67835), collected at Elizabeth River, 50 miles south of Darwin, Northern Territory, on 23 September 1960 by William Hosmer. Material examined. Specimen details are listed in the Appendix I. Type measurements. Hosmer (1964) presented a detailed description of the holotype (Fig. 9), but few measurements. Measurements (mm): SVL = 51.3, HL 19.4, HW = 18.1, TD = 3.9, ED = 4.7, EN = 5.3, IOD = 8.9, IND = 4.9, FLL = 12.6, Fin3L = 9.9, TL = 31.8, Toe4L = 27.3. Diagnosis. Morphologically shares the external features of L. watjulumensis (see above), but is distinguished from that species by divergence in mitochondrial and nuclear DNA markers (Figs 2, 3). From a genetic perspective, apomorphic nucleotide states at 12 and three sites in the mitochondrial ND4 and the nuclear PTPN12 genes respectively reliably diagnose L. spaldingi from L. watjulumensis (Table 5). Description including variation. Assessment of morphological variation is based on 16 females and 31 males (Table 2). SVL females mean = 47 mm, males mean = 38 mm. Head slightly longer than broad (HL/HW 1.17), and approximately one third snout to vent length (HL/SVL 0.39). Snout prominent, pointed when viewed from above blunt when viewed in profile. Nostrils more lateral than superior, closer to tip of snout than to eye. Distance between eye and naris equal to internarial span (EN/IND 1.03). Canthus rostralis well defined and straight. Eye size moderate, its diameter equivalent eye to naris distance. Pupil horizontal when constricted. Tympanum distinct, circular, length slightly greater than half eye diameter (TD/ED 0.62). Supratympanic fold absent or poorly developed. Vomerine teeth short straight plates bridging the gap between the choanae. Tongue approximately rectangular. Fingers long, slender, unwebbed. Subarticular and palmar tubercles prominent. Terminal discs not extending beyond lateral extremities of penultimate phalanx. Dark brown nuptial pad on upper and inner surface of the proximal half of the first finger. Fingers in order of length 3>4>1>2. Hindlimb length moderate (TL/SVL 0.67). Toes in order of length 4>5=3>2>1. Webbing reaches base of second most distal phalanx on toe 4 and penultimate phalanx on other toes. Subarticular tubercles prominent. Small oval inner metatarsal tubercle present. Terminal toe discs not wider than toes (Fig. 9). Dorsum either smooth or with low and infrequent tubercles sometimes forming lines parallel to long axis of the body. Limbs with low tubercules, sometimes smooth. Abdomen, undersurface of thighs, and lateral aspect of body mildly granular. Pectoral fold absent in a majority of specimens and when present is indistinct. Small vocal sac present. Vocal slit at base of mandible aligned along posterior-anterior axis, approximately one quarter length of mandible. Colour in life. Dorsum pale to light brown, mottled with dark brown, imparting an overall impression of a more-or-less uniformly light brown or dark brown animal (Fig. 10). In calling males yellow is suffused along the dorso-lateral edge of the dorsum from the eye to the groin and onto the lower back, onto upper forelimb and throat. Upper surfaces of limbs with same colour as dorsum, rarely with dark flecks. Face pale to light brown with dark brown stripe beginning at tip of snout and confined to upper edge of canthus rostralis, becoming more prominent after the nostril, continuing past the eye to mid-body with width same as eye and encompassing tympanum. In calling males, face often suffused with yellow. In some individuals, dark blotches extend from dorsal end of mid-lateral stripe to groin. Upper and lower lips either immaculate or have prominent light brown or white and dark brown patching. Dorsal surface of head often lighter than rest of dorsum in heavily patterned animals. Rear of thighs mostly dark brown, often same colour as facial stripe with numerous yellow and light brown circular to linear shaped spots and patches occupying between 20–50% of rear of thigh (Fig. 6). Yellow marks more frequent closer to knee, light brown marks dominate closer to vent. Vent same colour as surrounding dorsal colours. Fore of thighs either unpatterned or with same colour and pattern as rear of thigh but separated by intrusion of dorsal colour along dorsal surface of thigh. Groin with dark brown and yellow patching. Fore of lower limbs edged with dark brown in some individuals. Outer margin of foot well demarked by distinct border between lighter upper surface and dark brown plantar surface. Abdomen plain white, lower abdomen sometimes suffused with light brown flecking, otherwise immaculate. Throat white, sometimes with small indistinct dark mottling, otherwise unpigmented. Upper iris red margined with a distinct white band that appears continuous with upper margin of head stripe, lower iris same dark brown as head stripe. Distribution and habitat. Occurs in the IBRA regions of the Daly Basin, Darwin Coastal, Pine Creek, Arnhem Coast, Arnhem Plateau, Central Arnhem, Gulf Coastal, Gulf Fall and Uplands, Mount Isa Inlier, and Gulf Plains (Fig. 1). The most western locality is in the Litchfield National Park, NT (NTM R21723), and the most eastern and southern is the East Leichhardt River near the East Leichhardt Dam, Qld (SAMA R63646). One voucher, NTM R31013, a poorly preserved juvenile from the Murranji Stock Route, is the only record from the Sturt Plain bioregion. In Kakadu National Park and in the vicinity of Darwin, L. spaldingi is associated with riparian forests (Woinarski & Gambold 1992, Reynolds et al. 2010). Breeding biology. Tyler et al. (1983) provide a detailed account [as L. wotjulumensis] of the breeding biology, variation in egg and larval morphology and development of frogs from Magela Creek, a tributary of the East Alliga-tor River on the western periphery of the Arnhem Land plateau. Males called from the edge of water from October to March and breeding occurred early in the wet season. Spawn were found in temporary pools on sandy or gravelly soils. Advertisement call. Litoria spaldingi produces calls in long series that can last at least one minute (the maximum recording time of the FrogID app). Calls are similar to those of L. watjulumensis and exhibit similar levels of complexity and variation, but available recordings are dominated by short calls and long calls. These are similar to the short calls and long calls described above for L. watjulumensis. Clucks (see description of L. watjulumensis calls) were rarely recorded, but this may reflect a lack of social interactions between males on recorded sequences of L. spaldingi. Although some audio files contained only long or short calls, it is not possible to exclude the possibility that both call types were present before or after the limited segments on these audio files, because both call types occurred on several other recordings (Fig. 8B). Of 164 calls scored for call type, 135 (82%) are short calls and 29 (18%) are long calls. This difference is not attributable to sampling bias among contributors to FrogID because a similar ratio was observed on individual audio files containing both call types (e.g. Fig. 8B). Short calls are produced at intervals of 0.4–12.4 seconds (mean = 1.68, SD = 1.53, n = 96), and have a length of 0.29–0.69 seconds (mean = 0.47, SD = 0.07, n = 74). They typically comprise two discrete components: the first is a series of introductory notes produced at 0.05– 0.17 s intervals (mean = 0.09, SD = 0.03, n = 28). Only one of 78 short calls had no introductory notes (1.3%), four had one introductory note (5.1%), 14 had two introductory notes (18%), 55 had three introductory notes (70.5%) and four calls had four introductory notes (5.1%). Introductory notes are pulsatile, but amplitude modulation does not approach 100% except in rare cases (Fig. 8B). The second component of short calls is a rapidly-repeated series of ~7–12 terminal notes produced at much shorter intervals than short notes (approximately 0.006 – 0.010 s, but inter-note interval was difficult to measure accurately in most calls). A conspicuous feature of the terminal part of short calls is the rapid reduction in inter-note intervals across the course of this component. To the human ear short calls sound like a series of sharp clicks, followed by a short buzz during which note repetition rate increases rapidly. Terminal notes are also pulsatile, and there are some examples of 100% amplitude modulation producing distinct pulses. Amplitude of introductory notes is variable (Fig. 8B), but in terminal components amplitude starts very low and increases gradually, reaching maximum amplitude at the end of the call. Long calls are at least five times the duration of short calls (4.2– 12.2 s; mean = 8.2, SD = 2.2, n = 23), and lack introductory notes. Twenty-three long calls contained 56–129 notes (mean = 94.1, SD = 24.8) produced at a note repetition rate of 10.9–14.4 notes/s (mean = 12.9, SD = 1.1, n = 12). Amplitude is low at the start and increases gradually, typically reaching maximum amplitude approximately half to one third of the way through the call, then remains steady before dropping abruptly at the end of the call (Fig. 8B). However, one long call did not reach maximum amplitude until approximately three quarters of the way through the call. Notes are pulsatile but rarely achieve 100% amplitude modulation, although this is variable among calls. In some long calls each note is divided into two components: a pulsatile introductory component followed by 1–2 discrete pulses. Note repetition rate increases rapidly during the first ~20–25% of the call (Fig. 8B), and the call sounds to the human ear like a series of sharp clucks repeated increasingly rapidly and grading into a long, harsh chattering sound. Sixteen of 29 long calls terminate with 1–3 discrete notes reminiscent of short calls, or of the second component of short calls, i.e. a short ‘buzz’. Frequency is broadly distributed in both short and long calls (Fig. 8B) so the calls have a harsh quality. Dominant frequency of both call types is similar: 2490–3372 kHz (n = 19) in short calls and 2688–3007 kHz (n = 11) in long calls, but one long call had a dominant frequency that was carried by a poorly defined lower harmonic at 1540 kHz. Hoskin et al. (2015) present audio files of the advertisement calls of several males from Berry Springs and western Arnhemland. Comparisons between L. watjulumensis and L. spaldingi calls. The calls of L. watjulumensis and L. spaldingi are similar, and notable for their complexity. For convenience we have recognised three discrete call types: clucks, short calls and long calls. However, each of these call types exhibits variation in note structure within and between calls, and observations in the field indicate that some structural features of calls are adjusted by males according to social factors (Tyler & Doughty 2009). Given the unusually high variation in structural features of calls, and small sample sizes available for analysis (calls from just one male L. watjulumensis were available), robust comparisons between calls of L. watjulumensis and L. spaldingi were not feasible and consequently taxonomic implications of most differences observed are uncertain. One exception may be the striking difference in patterns of note rate adjustment during long calls. In long calls of L. watjulumensis, note repetition rate is relatively uniform during the first half of the call, then doubles abruptly (between two consecutive notes) and remains at the higher rate (with rare longer intervals between several notes) for the remainder of the call. In contrast, note rate in long calls of L. spaldingi gradually increase from the start of the call. Differences between these patterns are evident in Fig. 8. However, further studies are required to determine whether these patterns are consistent, and if so whether this feature of the species’ advertisement calls may be a useful diagnostic character in the field. Remarks. In his 1963 description of H. spaldingi, Hosmer cited Copland’s (1957) monograph but did not mention H. latopalmata watjulumensis, despite the latter occurring approximately 400 km WSW from the nearest record of his new species. Hosmer mentions examining 76 specimens of L. latopalmata, but provides no registration numbers or indication of the collection they were from (presumably AMNH). Based on their small adult body size, these were likely L. latopalmata from the eastern coast. Subsequently Tyler’s description of L. (Hyla) coplandi Tyler 1968, is the first time that adequate geographical representative material was used to assess morphological variation in L. watjulumensis sensu lato., Published as part of Donnellan, Stephen C., Catalano, Sarah R., Pederson, Stephen, Mitchell, Kieren J., Suhendran, Aidan, Price, Luke C., Doughty, Paul & Richards, Stephen J., 2021, Revision of the Litoria watjulumensis (Anura: Pelodryadidae) group from the Australian monsoonal tropics, including the resurrection of L. spaldingi, pp. 211-240 in Zootaxa 4933 (2) on pages 225-230, DOI: 10.11646/zootaxa.4933.2.3, http://zenodo.org/record/4550230, {"references":["Hosmer, W. (1964) A new frog of the genus Hyla from Northern Territory, Australia. American Museum Novitates, 2182, 1 - 7.","Woinarski, J. C. Z. & Gambold, N. (1992) Gradient analysis of a tropical herpetofauna: distribution patterns of terrestrial reptiles and amphibians in Stage III of Kakadu National Park, Australia. Wildlife Research, 19, 105. https: // doi. org / 10.1071 / WR 9920105","Reynolds, S. J., Dostine, P. L. & Griffiths A. D. (2010) Trial of a survey methodology to monitor frog assemblages in the Darwin region. Report number 26 / 2010 D. Northern Territory Government, Northern Territory of Australia. [unknown pagination, ISBN 978 - 1 - 921519 - 86 - 4]","Tyler, M. J., Crook, G. A. & Davies, M. (1983) Reproductive biology of the frogs of the Magela Creek System, Northern Territory. Records of the South Australian Museum, 18, 415 - 440.","Hoskin, C., Grigg, G., Stewart, D. & McDonald, S. (2015) Frogs of Australia-a complete electronic field guide to Australian frogs. Ug Media, Australia. Available from: ugmedia. com. au / field-guides / frogs-of-australia / (accessed 19 January 2021)","Tyler, M. J. & Doughty, P. (2009) Frogs of Western Australia. 4 th Edition. Western Australian Museum Press, Perth, 160 pp.","Copland, S. J. (1957) Australian tree frogs of the genus Hyla. Proceedings of the Linnean Society of New South Wales, 82, 9 - 108.","Tyler, M. J. (1968) A taxonomic study of hylid frogs of the Hyla lesueuri complex occurring in northwestern Australia. Records of the South Australian Museum, 15, 711 - 727."]}

Litoria watjulumensis (Copland, 1957) Wotjulum Frog (Figs 5, 6, 7) Hyla latoplamata watjulumensis Copland, 1957 Lectotype. WAM R 11896, an adult female, collected at Wotjulum, Western Australia (16°11’ S, 123°37’ E) by A.M. Douglas. Material examined. Specimens from WA and NT are listed in the Appendix I. Lectotype measurements (mm): SVL = 46.6, HL = 19.0, HW = 16.1, TD = 3,5, ED = 5.1, EN = 5.1, IOD = 8.0, IND = 4.5, FLL = 11.3, Fin3L = 7.2, TL = 31.7, Toe4L = 22.3. Diagnosis. Distinguished from other Australian Litoria (except L. spaldingi) by a combination of: vomerine teeth present; distinct tympanum; strong head stripe, covering tympanum; grey to brown colouration in life, with dark variegations and infused with yellow which is prominent in males (Fig. 5); black shin; inner thigh colouration comprises variously shaped yellow to orange or light brown spots on a darker brown to black background (Fig. 6); fingers without webbing; fully webbed feet; and moderately expanded terminal discs on all digits (Fig. 7), and a complex advertisement call that comprises sporadic chicken-like clucks, followed by a long series of rapid evenly spaced notes followed by more clucks. Morphologically shares the external features of L. spaldingi and has a similarly structured advertisement call, but is distinguished from that species by divergence in mitochondrial and nuclear DNA markers (Figs 2, 3). From a genetic perspective, apomorphic nucleotide states at nine and three sites in the mitochondrial ND4 and the nuclear PTPN12 genes respectively reliably diagnose L. watjulumensis from L. spaldingi (Table 5). A) B) Description including variation. Assessment of morphological variation is based on 35 females and 39 males (Table 2). Mean SVL: females = 49 mm, males = 36 mm. Head slightly longer than broad (HL/HW 1.24) and approximately one-third of SVL (HL/SVL 0.39). Snout prominent, blunt when viewed from above and in profile. Nostrils more lateral than superior, closer to tip of snout than to eye. Distance between eye and naris equal to internarial span (EN/IND 1.03). Canthus rostralis well defined and straight. Eye relatively large, its diameter greater than eye to naris distance. Pupil horizontal when constricted. Tympanum distinct, circular, length slightly greater than half eye diameter (TD/ED 0.66). Vomerine teeth short straight plates bridging the gap between the choanae. Tongue approximately rectangular. Fingers long, slender, unwebbed. Subarticular and palmar tubercles prominent. Terminal discs not wider than fingers. Dark brown nuptial pad on upper and inner surface of the proximal half of the first finger. Fingers in order of length 3>4>1>2. Hindlimb length moderate (TL/SVL 0.64). Toes in order of length 4>5=3>2>1. Webbing reaches base of second most distal phalanx on toe 4 and penultimate phalanx on other toes. Subarticular tubercles prominent. Small oval inner metatarsal tubercle prominent. Terminal toe discs not wider than toes. Dorsum either smooth or with low and infrequent tubercules sometimes forming lines parallel to long axis of the body. Limbs with low tubercules, sometimes smooth. Abdomen, undersurface of thighs, and lateral aspect of body mildly granular. Pectoral fold absent in a majority of specimens and when present is indistinct. Vocal slit at base of mandible aligned along posterior-anterior axis, approximately one third length of mandible. Colour in life. Dorsum pale to light brown, often heavily mottled with dark brown and in some individuals additional red mottling, imparting an overall impression of a more-or-less uniformly light or dark brown animal (Fig. 5). Upper surfaces of limbs with same colour as dorsum, rarely with dark flecks. In calling males, yellow is suffused along the dorso-lateral edge of the dorsum from the eye to the groin and onto the lower dorsum. Head pale to light brown with dark brown stripe beginning at tip of snout becoming more prominent after the nostril but confined to upper edge of canthus rostralis, continuing past the eye to mid-body, with width same as eye and encompassing tympanum. In calling males, head often suffused with yellow. In some individuals, dark blotches from dorsal end of side stripe to groin. In all individuals lower lip has prominent light and dark brown patching. For some individuals the upper lip has the same patterning as the lower lip, otherwise upper lip is immaculate. Dorsal surface of the head often lighter than rest of the dorsum due to reduced frequency and extent of darker mottling. Rear of thighs mostly dark brown, often same colour as head stripe with numerous yellow, circular to linearshaped spots and patches occupying between 20–70% of the rear of the thigh (Fig. 6). Vent same colour as surrounding dorsal colours. Fore of thighs and groin with same colour and pattern as rear of thigh but separated from rear thigh pattern by intrusion of dorsal edge of thigh. Outer margin of foot with lighter upper surface distinctly bordered by dark brown of plantar surface. Abdomen plain white, lower abdomen sometimes suffused with light brown flecking, otherwise immaculate. Throat sometimes with small indistinct dark brown mottling otherwise unpigmented. Upper iris red margined with a distinct white band that appears continuous with upper margin of facial stripe, lower iris same dark brown colourations as facial stripe. Distribution and habitat. Occurs in the IBRA (Interim Biological Regionalisation of Australia) regions of Dampierland, Northern Kimberley, Central Kimberley, Victoria Bonaparte, Ord Victoria Plain, and Tanami (Fig. 1). The most western locality is the Stewart River, Kimbolton (WAM R51854), the most eastern is in the Victoria River catchment in Gregory National Park (NTM R37297 –8), and the most southern is Slatey Creek in the Sturt Creek catchment in the north-western Tanami (WAM R132982). Breeding biology. Francis (2013) described the breeding biology from the east Kimberley region of Western Australia where it breeds in elevated rocky pools early in the wet season. Anstis (2013) presented a combined description of variation in embryos and larvae of L. watjulumensis and L. spaldingi, only noting where particular individual sampled localities departed in individual trait states from the remainder. However, Anstis did not note any difference between groups of the sites that we could allocate to L. spaldingi vs L. watjulumensis based on geography. Advertisement call. Tyler & Doughty (2009) describe the call of L. watjulumensis as very complex, comprising sporadic chicken-like clucks, followed by a rapid series of evenly spaced notes (3–4 per second) which abruptly double in rate without a pause for up to 30 seconds followed by more clucks. The clucks are typically given in response to other male calls (Tyler & Doughty 2009). Hoskin et al. (2015) present an audio file of the advertisement call of a male from Kununurra. Our analysis is based on a sequence of 100 calls from one male (WAM R162508) of 165 s duration. Calls are complex and there is substantial variation in structural components within the sequence. However, calls can be grouped for convenience into three major types: 1) ‘clucks’ (53%), 2) ‘short calls’ (39%) and 3) ‘long calls’ (8%). Clucks are sharp notes produced singly or in short bursts, with each note lasting 0.02– 0.04 s (mean = 0.03, SD = 0.005, n = 38). Clucks may be pulsatile, with the extent of amplitude modulation of pulses, including the proportion of pulses achieving 100% amplitude modulation, varying both within and between clucks; or they may comprise a single pulse. Short calls last 0.35– 1.00 s (mean = 0.48, SD = 0.13, n = 30) and comprise two discrete components. The first is a series of 3–8 (mean = 3.73, SD = 1.16, n = 41) slowly repeated introductory cluck notes produced at intervals of about 0.04– 0.05 s, although this is variable. Twenty-two (53.7%) short calls have three introductory notes, and 15 (36.6%) have four introductory notes; just four short calls have between five and eight introductory notes. Introductory notes are pulsatile and frequently exhibit 100% amplitude modulation between 2–3 distinct pulses; the final pulse is normally significantly longer than preceding pulses. The second component of short calls is a rapidly repeated series of about 4–7 terminal notes produced at short intervals that are difficult to measure accurately. Terminal notes are finely pulsed and in total this second component of short calls lasts 0.13– 0.19 s (mean = 0.16, SD = 0.02, n = 28). Note repetition rate does not shift noticeably during the second component of short calls. Short calls sound like a series of clucks followed by a short, harsh buzz. Long calls are a train of rapidly repeated notes that is at least eight times longer than short calls (8.6– 16.4 s; mean = 12.1, SD = 2.8, n = 8). Eight long calls contain 82–142 notes (mean = 109.4, SD = 20.7) produced at an overall repetition rate of 8.4–10.0 notes/s (mean = 9.1, SD = 0.6). However, although note rate does not change noticeably during the first half of the call, it nearly doubles from 6.7–7.1 notes/s (mean = 6.91, SD = 0.19) to 11.5–13.2 notes/s (mean = 12.57, SD = 0.54) at a sharp boundary (between two consecutive notes) between 48.2 and 71.0% of the way through the call (mean = 61.70%, SD = 9.08) (Fig. 8A). Amplitude is low at the start and increases rapidly for the first ~10-15% of the call then more gradually until it reaches maximum amplitude near the end of the call. Notes are very finely pulsed, and there may be a ‘supplementary’ terminal pulse that is separated from preceding pulses by about 0.004 s, although this is variable among calls. Notes within long calls are similar to cluck calls but are generally longer. The length of 20 notes measured from before, and 20 from after, the switch from ‘slow’ to ‘fast’ note rate in the first recorded long call of WAM R162508 are 0.04– 0.05 s (mean = 0.048, SD = 0.004, n = 20) and 0.04- 0.5 s (mean = 0.049, SD = 0.003, n = 20), versus note length of 0.02– 0.04 s in cluck calls. Frequency is broadly distributed in the three call types (Fig. 8A) so they have a harsh quality. Dominant frequency of the call types overlaps broadly: 2760–3210 kHz (mean = 3023, SD = 154.4, n = 15) in clucks, 2731–3121 kHz (mean = 2981, SD = 97.92, n = 28) in short calls and 2857–3002 kHz (n = 8) in long calls. Figure 8 illustrates a long call followed by a burst of clucks, then two short calls, five clucks, and 18 short calls followed by nine clucks., Published as part of Donnellan, Stephen C., Catalano, Sarah R., Pederson, Stephen, Mitchell, Kieren J., Suhendran, Aidan, Price, Luke C., Doughty, Paul & Richards, Stephen J., 2021, Revision of the Litoria watjulumensis (Anura: Pelodryadidae) group from the Australian monsoonal tropics, including the resurrection of L. spaldingi, pp. 211-240 in Zootaxa 4933 (2) on pages 220-225, DOI: 10.11646/zootaxa.4933.2.3, http://zenodo.org/record/4550230, {"references":["Copland, S. J. (1957) Australian tree frogs of the genus Hyla. Proceedings of the Linnean Society of New South Wales, 82, 9 - 108.","Francis, J. (2013) Ecological studies of anuran larvae in temporary freshwaters of the Australian wet-dry tropics. PhD Thesis, Scholl of Animal Biology, University of Western Australia, Perth, xiii + 236 pp.","Tyler, M. J. & Doughty, P. (2009) Frogs of Western Australia. 4 th Edition. Western Australian Museum Press, Perth, 160 pp.","Hoskin, C., Grigg, G., Stewart, D. & McDonald, S. (2015) Frogs of Australia-a complete electronic field guide to Australian frogs. Ug Media, Australia. Available from: ugmedia. com. au / field-guides / frogs-of-australia / (accessed 19 January 2021)"]}

Cyclorana platycephala (G��nther, 1873) Eastern Water-holding Frog Figs. 5���9 Chiroleptes platycephalus G��nther, 1873 Holotype: BMNH 1947.2. 18.42, adult female collected at Fort Bourke, NSW, Australia. Synonymy (by Tyler 1990) Cyclorana slevini Loveridge, 1950 Material examined. Details of the 143 specimens used to document variation are presented in Appendix 1. Holotype meristics. G��nther (1873) presented a detailed description of the holotype but few measurements. We include images of the holotype (Fig. 6) and comprehensive set of measurements (mm): SVL = 52.9, FA = 12.8, HW = 25.5, HL 22.4, IO = 9.3, IN = 3.0, ED = 3.5, EN = 4.4, SN = 5.6. Redefinition of Cyclorana platycephala Diagnosis. A medium to large robust burrowing species (SVL: males 43���63 mm, females 47���72 mm). Clearly assigned to the Cyclorana by its phylogenetic placement in molecular genetic analyses (Figs. 2) and a combination of its morphological characters, burrowing behaviour, tadpoles and call structure. Distinguished from all other congeners, except C. occidentalis, by the more dorsally-tilted eyes, fully webbed toes (no more than half-webbed in all other Cyclorana), and the oral disc of tadpoles (Anstis 2013). For distinction from C. occidentalis see Diagnosis for C. occidentalis below. Variation. See also morphological measurements of all preserved specimens (Appendix 1; Table 2) and details of morphological variation (Table 3). Head broad, somewhat depressed, and wider than long. Snout short, rounded to truncate in dorsal view and in profile, mouth wide. Eyes prominent and dorsally oriented, diameter greater than eye-to-nostril distance, with some difference noted in the northern populations (average ED/EN males/females: eastern populations = 1.36 �� 0.18; northern 1.6 �� 0.24). Nostrils open mainly dorsally, with slight lateral tilt, slightly closer to tip of snout than to eyes, distance between them very slightly less than eye to nostril (average IN/EN 0.92 �� 0.12). Canthus rostralis curved, or slightly so in anterior and lateral views (Fig. 7), may be almost nonexistent in some very short-snouted northern specimens. Slight ridge from eye to nostril in some. Tympanum distinct, diameter less than eye diameter (average Tymp/ED = 0.80 �� 0.07); annulus slightly raised, supra-tympanic skin fold distinct or indistinct and extends from edge of eye, above tympanum and around its posterior rim, terminating above forearm. Toes fully webbed, fingers unwebbed, thumb opposable. Dorsal skin mostly smooth with or without granular sides, scattered low tubercles present in some and occasionally a few short skin folds (see Table 3). Ventral surface smooth. Tongue very broad, attached anteriorly, free behind. A prominent vomerine tooth plate on either side of midline, each narrowly separated and laterally adjacent to medial edge of choanae; each tilts slightly posteriorly towards medial edge; ventral edge denticulate with 5���7 small pointed tips; choana almost equal in width to vomerine tooth plate (Fig. 8 A,B). Arms fairly short (FA/SVL = 0.23 �� 0.08), hindlimbs short and robust (TibL/SVL 0.39 �� 0.04). Fingers unwebbed, decreasing in order of length 3 �� 4 �� 1 �� 2; subarticular tubercles well defined, inner metacarpal tubercle large, oval and fleshy, tapering anteriorly; toes decreasing in order of 4 �� 5 �� 3 �� 2 �� 1, fully webbed to tips but tapering to broad lateral fringes towards tip of fourth toe; subarticular tubercles poorly defined and small, inner metatarsal tubercle large, ovoid and partly or fully unattached beneath medial edge; tips of fingers and toes slightly fleshy, no wider than digit and no circum-marginal grooves (Fig. 8 G,H). Colour and skin texture in life. Dorsum ranges from grey, often with green areas and sometimes pinkish tinges, to dull yellow-brown (with or without darker flecks or patches) or uniform green. Specimens from the grey and brown clay soils of the Bourke/Nyngan/Gilgandra regions in western NSW usually have smooth skin, and adults tend to have a grey dorsum with green areas (with or without scattered flecks or pinkish tinges) or are sometimes all dull green. Specimens from south-western Qld are dull yellow-brown with or without reddish tinges (Fig. 9). Some specimens have a paler green or cream vertebral stripe (Fig. 9 G). Limbs and upper lip usually with darker flecks or mottling. Further north in the Boulia region in south-western Qld, the skin is more granular, with scattered tubercles, and frogs in life tend to be dull yellow-brown (some with dull reddish tinges). Specimens found in the heavy grey clay soil areas from the north-eastern locality east of Clermont, Qld are immaculate light grey with faint greenish tinges and smooth skin (Meyer & Agnew 2013). Those from the Barkly Tableland, NT mostly have smooth grey skin (29 % slightly granular), with a pink wash over the eyelids, along either side of the vertebral region and on the dorsal surface of the limbs (Fig. 9 H, K, N), and some have scattered olive-grey flecks (Fig. 9 B, E). Belly white; throat dark grey in calling, adult males. Iris golden. Colour in preservative. Dorsal colour grey, grey-brown or brown, with or without faint darker flecks or mottling; some specimens (3 %) with pale-grey vertebral stripe. No specimens from the northern populations have a pale vertebral stripe. Limbs and upper lip usually mottled or flecked. Belly cream or white, throat dark grey in adult males. Tadpoles. Tadpoles were described by Anstis (2013). Photographs in life of a specimen from eastern and northern regions and morphometric measurements are provided here for comparison (Fig. 10; Table 4). Maximum size: TL to 109 mm, BL to 37 mm (stage 41, Bourke, NSW), but many only grow to about 70���80 mm in total length. Tadpoles of C. platycephala have moderately large to very large, plump bodies with dorsolateral eyes and moderately arched fins. Colour in life. Tadpoles in earlier stages 25���26, as observed in the field from an eastern population, have a fine layer of gold over dorsum and may have a darker line along either side of vertebral region with a darker area over base of body. As they grow the dorsum becomes more uniformly pigmented with a layer of gold, merging with opaque silver-white and a copper-gold or greenish sheen over sides of body and venter. Iris gold. Tail muscle in fully-grown tadpoles light or darker grey-brown, with gold flecks over sides. Fins transparent to dusky dark grey, densely speckled with gold and often green iridophores. Tadpoles from northern populations on the Barkly Tableland were only observed in later stages, but were dark olive-brown over the dorsum merging with a green sheen and dull opaque silver-white over sides and on venter. Colour in preservative. All gold and silver-white pigment is lost in preservative and tadpoles are dull and more translucent with darker internal organs visible. Oral disc. The key characters summarised from Anstis (2013) are: broad anterior medial gap in border of marginal papillae, two anterior and three posterior tooth rows with medial gap in second anterior and first posterior rows, robust jaw sheaths, upper broadly arched. Metamorphosis. The duration of larval life ranges from 30���50 days (Anstis 2013), but in hotter regions in summer may be much less. Metamorphosis has been observed in late October to May in eastern populations from NSW (spring to autumn), and in late summer (February) in the Barkly Tableland, NT, but in areas of low and unpredictable rainfall, could occur at any time from spring to autumn. Metamorphs strongly resemble the adult in colour and skin texture, but the tympanum is not distinct initially. Fourteen recent metamorphs from tadpoles raised in captivity from eastern populations had a mean SVL of 27 mm (22.0���38.0 mm) and two from the Barkly Tablelands were 26.0 and 25.5 mm (Anstis 2013). SVL of recent metamorphs observed in the field were similar to those raised in captivity, but none were measured. Habitat. Tadpoles of C. platycephala have been observed in muddy, grey or brown water and clearer ephemeral water bodies in clay-soil areas, where they tend to be whitish-gold (muddy water) or yellow-ochre (clearer water) when fully grown, with a silver-white belly. Some have also been observed in the field with black tails at night and early morning (Meyer & Agnew 2013; M. Anstis, unpubl. data).Those at more advanced stages from the Barkly Tableland were dull grey to olive-brown above in clearer water over black, clay substrate. Etymology. The specific epithet is Latin for ���flat��� (platy) and ���headed��� (cephala). Distribution and habitat. Occurs across a wide area of eastern Australia in lower rainfall inland areas of NSW, Qld, southern NT and northern SA (Fig. 1). There are specimens from more than 10 Interim Biogeographic Regionalisation of Australia (IBRA) bioregions (Thackway & Cresswell 1995; Department of Environment 2012), but principally, in decreasing order of number of specimens, from: Channel Country, Mulga Lands, Darling Riverine Plains, Stony Plains, Cobar Peneplain and Riverina Bioregions (Atlas of Living Australia website www.ala.org.au, accessed 1 Sep 2015). They commonly occur in clay-based soils where water is more likely to be held for longer periods at the surface after heavy rain. Within the Brigalow Belt, for example, they avoid sandy cypress and sandstone areas in favour of adjoining communities such as brigalow, belah and coolabah on clay soil (H.B. Hines & E. Meyer, unpubl. data). After active periods, they burrow into clay soil, where they remain cocooned in their sloughed skin within a small cavity, sometimes for several years (van Beurden 1982). Advertisement call. The call is a long continuous pulse train, rising in amplitude mid-call, then declining relatively gradually, resulting in a nasal ��� maaaaaaaah��� repeated every 1���2 seconds (Fig. 5; Table 5). Breeding behaviour. An explosive breeder, with breeding occurring in ephemeral wetlands within a few hours of heavy rainfall in warmer months. Males call while partly submerged or afloat in water. Calling, and presumably breeding, drops off considerably or ceases within a day or two of the rainfall event. In NSW, Lemckert & Mahony (2008) showed calling records for this species in October, November, March and April. In Qld, calls have been heard in November, December, February and March, with amplectant pairs observed in February (H.B. Hines, unpubl. data). Small tadpoles at stage 25 have been collected in October (M. Anstis, unpubl. data). From the detailed studies on the natural history of C. platycephala undertaken in the Coonamble-Walgett district of New South Wales (van Beurden 1982), breeding does not seem to occur following heavy rainfall in winter., Published as part of Anstis, Marion, Price, Luke C., Roberts, J. Dale, Catalano, Sarah R., Hines, Harry B., Doughty, Paul & Donnellan, Stephen C., 2016, Revision of the water-holding frogs, Cyclorana platycephala (Anura: Hylidae), from arid Australia, including a description of a new species, pp. 451-479 in Zootaxa 4126 (4) on pages 463-469, DOI: 10.11646/zootaxa.4126.4.1, http://zenodo.org/record/271746, {"references":["Gunther, A. (1873) Description of two new species of frogs from Australia. Annals of the Magazine of Natural History, Series 4, 11, 349 - 350. http: // dx. doi. org / 10.1080 / 00222937308696827","Tyler, M. J. (1990) Geographic distribution of the fossorial hylid frog Cyclorana platycephala (Gunther) and the taxonomic status of C. slevini Loveridge. Transactions of the Royal Society of South Australia, 114, 81 - 85.","Loveridge, A. (1950) New frogs of the genera Cyclorana and Hyla from south-eastern Australia. Proceedings of the Biological Society of Washington, 63, 131 - 138.","Anstis, M. (2013) Tadpoles and frogs of Australia. New Holland Publishers, Sydney, 832 pp.","Meyer, E. & Agnew, L. (2013) A range extension of the water holding frog (Cyclorana platycephala) in Queensland. Queensland Naturalist, 51, 19 - 22.","Thackway, R. & Cresswell, I. D. (1995) An interim biogeographic regionalisation of Australia. Australian Nature Conservation Agency, Reserve Systems Unit, Canberra, 88 pp.","van Beurden, E. (1982) Desert adaptations of Cyclorana platycephalus: a holistic approach to desert adaptation in frogs. In: Barker, W. R. & Greenslade, P. J. M. (Eds.), Evolution of the flora and fauna of Arid Australia. Peacock Publications, South Australia, pp. 235 - 240.","Lemckert, F. & Mahony, M. (2008) Core calling periods of the frogs of temperate New South Wales, Australia. Herpetological Conservation and Biology, 3, 71 - 76."]}

Cyclorana occidentalis sp. nov. Western Water-holding Frog Figs. 6���9 Holotype. WAM R 111826, adult female collected 55 km E Newman, Pilbara, WA (23.4908 ��S, 120.3172 ��E) by P. Doughty on 4 October 2005. Material examined. Details of the 41 specimens used to document variation are listed in Appendix 1. Diagnosis. A medium to large robust burrowing species, SVL males 42���60 mm; females 44���66 mm. Clearly assigned to the Cyclorana by its phylogenetic placement in molecular genetic analyses (Fig. 2) and a combination of its morphological characters, burrowing behaviour, tadpoles and call structure. Distinguished from all other species of Cyclorana except C. platycephala by the more dorsally-tilted eyes, fully webbed toes (no more than half-webbed in all other Cyclorana) and the oral disc of tadpoles (Anstis 2013). Distinguished from C. platycephala by its western Australian distribution (Fig. 1), the consistently greater number of tubercles and skin folds, yellow-brown dorsal colour (never green or pink as in C. platycephala), by molecular genetic analyses and the call. The call of C. occidentalis increases in amplitude steadily over the first �� of the call, compared with amplitude peaking about mid-call in C. platycephala). The only species of Cyclorana sympatric with C. occidentalis is C. maini from which C. occidentalis can be separated by a combination of larger body size, fully webbed toes (one-third webbed in C. maini), more prominent dorsal tubercles and skin folds, broader head with more dorsally-tilted eyes, dorsum rarely with numerous darker patches (as in C. maini), oral disc of the tadpoles and the call. Calls of C. maini, as described by Tyler & Martin (1976), have a higher pulse rate (244 pulse.s - 1), higher frequency (1922 Hz) and the call is longer (814 ms) than the call of C occidentalis. Description of holotype. Moderately large and robust in habitus (SVL 64 mm). Head broad, slightly flattened and slightly wider than long (HW/HL = 1.06); snout broadly rounded in dorsal view, obtusely rounded in profile; eyes prominent and dorsolateral in position, inclined anterolaterally with a dorsal tilt; eyelids granular; transverse diameter slightly greater than eye-to-nostril distance (ED/EN = 1.12). Canthus rostralis poorly defined and very slightly curved in anterior and lateral views (Fig. 6 A, B); nostrils slightly closer to tip of snout than to eye, distance between them less than eye to nostril (IN/EN 0.74); tympanum distinct, in anterior view projecting dorsolaterally, annulus slightly raised, diameter slightly less than eye (TYMP /ED = 0.96); supratympanic fold distinct and sloping ventrally posterior to tympanum, terminating above forearm. Tongue very broad, attached anteriorly and free behind; a prominent vomerine tooth plate either side of midline, each laterally adjacent to medial edges of choanae and narrowly separated; each tilting slightly posteriorly towards medial edge; ventral edges denticulate, with four to six small pointed tips; choana about equal in length to the width of a vomerine tooth plate; mouth could not be opened wide enough to photograph, so Fig. 8 shows features of mouth of WAM R 165306, which agrees with description of holotype above. Arms short (FA/SVL = 0.21); hindlimbs short and robust (TL/SVL = 0.37). Tips of fingers and toes slightly fleshy, no wider than digit, no circum-marginal grooves; fingers slender and unwebbed, decreasing in length in order 3 �� 4 �� 1 �� 2; thumb opposable; inner metacarpal tubercle large and prominent, palmar tubercle broad and flattened; subarticular tubercles distinct and projecting, especially on first finger; toes slender, decreasing in length in order of 4 �� 5 �� 3 �� 2 �� 1 and fully webbed to tips, tapering into broad lateral fringes towards tip of longest (fourth) toe; inner metatarsal tubercle prominent, shovel-shaped and medial edge free of attachment to foot beneath; outer metatarsal tubercle indistinct, broad and flat; subarticular tubercles very small and indistinct. Dorsum with numerous smoothly rounded, low tubercles and a few short skin folds; a long dorsolateral skin fold extends from posterior of tympanum to more than midway along flanks; eyelids granular; ventral surface granular. Colour in life. Dorsum yellow-ochre with diffuse, faintly darker yellow and grey flecks, increasing posteriorly and over legs (Fig. 9). Dorsal colour merges into cream over sides of body. Undersurface white. Iris golden. Colour in preservative. All yellow pigment is lost; dorsum grey, eyelids with subdued darker spots, eye to tympanum darker; limbs and flanks mottled; undersurface cream with light grey spots on throat. Variation. Details of all specimens examined for this description of variation are presented in Appendix 1. See also morphological measurements of all preserved specimens (Table 2) and details of morphological variation (Table 3). SVL males 42���60 mm, females 46���70 mm. TibL/SVL range is 0.34���0.43 (mean 0.39 �� 0.02). The dorsal skin of all but one frog (WAM R 154929) bears tubercles to varying degrees (dense in 51 %), and all but one (WAM R 63834) have a long dorsolateral skin fold beginning posterior to the tympanum. Most (76 %) have additional short skin folds (Table 3). Ventral skin smooth (23 %) to granular (77 %). The snout is mostly rounded in dorsal view and in profile, 38 % are truncate in dorsal view and in profile. The canthus rostralis is slightly curved or curved in anterior and lateral views (Fig. 7 I, M), with a slight ridge from eye to naris in some. Tadpoles. Maximum size in life: TL to 82 mm, BL to 32 mm (stage 39, Carnarvon, WA). Tadpoles of C. occidentalis are generalised pond-dwellers and have a moderately large to large, plump body with moderately arched fins (Anstis 2013). Photographs of tadpoles in life and a drawing of the oral disc and morphometric comparisons are given in Fig. 9 and Table 3. As tadpoles have a similar morphology to those of C. platycephala, no drawing of the tadpole is included. Photographs of tadpoles in life and a drawing of the oral disc are included (Fig. 10), and morphometric comparisons of 10 tadpoles of C. occidentalis with samples of C. platycephala from eastern and northern populations (Table 4; Fig. 4). Colour in life. Tadpoles in earlier stages 25���26 are translucent gold with darker areas over naris to eye, brain and vertebral region. Dorsal tail muscle with alternating gold and black patches anteriorly (when first collected). As tadpoles grow, yellow-gold pigment increases in coverage and density over dorsum and merges into dense, opaque silver-white over sides of body and venter. Iris dense gold. Tail muscle in fully-grown tadpoles light golden, fins transparent, densely speckled with gold. Colour was similar in all specimens raised or freshly collected. Colour in preservative. All gold and silver-white pigment is lost in preservative and tadpoles are dull and translucent with darker internal body areas visible. Oral disc. As for C. platycephala, but the jaw sheaths may be slightly less robust. Metamorphosis. Recorded in January, but probably occurs at any time from January to March in north-western Australia, depending on the timing of summer rains. Metamorphs strongly resemble the adult in colour and skin texture (Fig. 10), but the tympanum is not distinct initially. Four had a mean SVL of 30 mm (26.0���32.0). Observations of recent metamorphs in the field indicated the SVL was relatively similar to those raised in captivity, but none was measured. Habitat. The light yellow-golden colour of C. occidentalis tadpoles found in the Wooramel and Carnarvon districts of WA is consistent with the yellow-brown opaque muddy water of the large claypans and floodplains where they occurred. Comparisons with tadpoles of other species. Tadpoles of C. occidentalis could be confused with those of C. maini and Neobatrachus because of similarities in maximum size and colour. Cyclorana occidentalis tadpoles have less heavily keratinised jaw sheaths (robust) with a more broadly arched upper jaw sheath than those of C. maini, which have very heavily keratinised jaw sheaths (massive) and a more narrowly arched upper jaw sheath (Anstis 2013). Neobatrachus tadpoles differ from Cyclorana in having three or four anterior tooth rows (two in Cyclorana), dorsal eyes and massive jaw sheaths. From stage 38, C. occidentalis have more extensive webbing between the toes than any of the other sympatric species above. Etymology. The species epithet is Latin for ���western��� and refers to its distribution in the western third of the Australian continent. Distribution and habitat. This species inhabits arid or semi-arid areas of central WA, south from about Karratha in the north to the Kalgoorlie area, and from the coast inland to Well 26 on the Canning Stock Route in the north and Mavis Rock in the south (Fig. 1). Most specimens are from the Pilbara, Murchison, Gascoyne, Yalgoo and Carnarvon IBRA regions (Department of the Environment 2012; Atlas of Living Australia website at www.ala.org.au �� accessed 1 Sep 2015). Several specimens from Barrow Island, eastern WA and the south-western and north-western NT fall outside of the distribution we present in Fig. 1. WAM R 40026 from Barrow Island could not be located in the WAM collection, and with no subsequent sightings or calls of this species recorded from the island despite extensive survey work, we view this record as suspect. NTM R 30138 from the ���Victoria River District��� could not be located at the NTM so we have not been able to confirm its identity. We assessed specimens attributed to ��� C. platycephala ���, WAM R 21320 from Warburton, WAM R 21535 and R 96360 ��� 8 from Palm Valley, AMS R 21117 from 240 km W Alice Springs, but all are either C. maini Tyler & Martin or Platyplectrum spenceri Parker. These records were not included in Fig. 1. Based on records of frogs from museum collections viewable on line (Atlas of Living Australia 2015), the absence of C. occidentalis or C. platycephala from the intervening area of these species ranges is not readily explained by lack of sampling, as other arid frogs such as Neobatrachus Peters, Notaden G��nther, Platyplectrum spenceri and other Cyclorana species are recorded from much of this area. Therefore we view the geographic separation between taxa to be real, possibly caused by differences in soil type or historical factors, for example, relatively recent range expansion, as evidenced by lack of mitochondrial phylogeographic structure within C. occidentalis, and C. verrucosa relative to that observed in C. platycephala (Fig. 2). Within its broad range in the western deserts, C. occidentalis appears to have a preference for areas with heavy clay soils and has been encountered in ephemeral pools along creek lines. It avoids desiccation by aestivating underground and forming a cocoon of sloughed skin (Withers 1995). Advertisement call. The call is a nasal ���waaaaaaaaah��� increasing in amplitude over the first �� of the call, decreasing rapidly after that, and repeated ��� every 1.5 seconds���often in long calling sequences (see details Table 5, Fig. 5). A call can be heard at: http://museum.wa.gov.au/explore/frogwatch/frogs/water-holding-frog. Breeding behaviour. MA observed males and gravid females at night beside water bodies at Carnarvon WA on 12 January 2011 and also found tadpoles in stages 26���28 at this time. JDR observed calling males in a pool in a drainage line following heavy rains near Meekatharra, December 19 ���20, 1988 where calls were recorded., Published as part of Anstis, Marion, Price, Luke C., Roberts, J. Dale, Catalano, Sarah R., Hines, Harry B., Doughty, Paul & Donnellan, Stephen C., 2016, Revision of the water-holding frogs, Cyclorana platycephala (Anura: Hylidae), from arid Australia, including a description of a new species, pp. 451-479 in Zootaxa 4126 (4) on pages 469-472, DOI: 10.11646/zootaxa.4126.4.1, http://zenodo.org/record/271746, {"references":["Anstis, M. (2013) Tadpoles and frogs of Australia. New Holland Publishers, Sydney, 832 pp.","Tyler, M. J. & Martin, A. A. (1976) Taxonomic studies of some Australian leptodactylid frogs of the genus Cyclorana Steindachner. Records of the South Australian Museum, 17, 261 - 276.","Withers, P. C. (1995) Cocoon formation and structure in the aestivating Australian desert frogs, Neobatrachus and Cyclorana. Australian Journal of Zoology, 43, 429 - 441. http: // dx. doi. org / 10.1071 / ZO 9950429"]}

Anstis, Marion, Price, Luke C., Roberts, J. Dale, Catalano, Sarah R., Hines, Harry B., Doughty, Paul, Donnellan, Stephen C. (2016): Revision of the water-holding frogs, Cyclorana platycephala (Anura: Hylidae), from arid Australia, including a description of a new species. Zootaxa 4126 (4): 451-479, DOI: http://doi.org/10.11646/zootaxa.4126.4.1

Characterization of the mobility-lifetime product is critical to the development of new materials for semiconductor radiation detectors. An optical technique has been developed that allows for the direct determination of the minority carrier diffusion length, drift length, and mobility-lifetime product from a single image of the recombination luminescence in semiconductor materials. Excess carriers are generated using the electron beam in a scanning electron microscope. The charge is then drifted by applying an electric field, and the subsequent recombination luminescence is imaged by an optical microscope on a high-sensitivity CCD camera. The challenge in applying this technique to new materials for nuclear radiation detectors is the requirement for thick samples and the resultant need to characterize transport in three dimensions. In this work, initial research was performed on the simulations and analysis of experimental data required to characterize thick layers of high purity GaAs for nuclear radiation detectors. The first models were applied to extract values for surface recombination velocity, which plays a key role in determining the excess carrier distribution in bulk materials. Cathodoluminescence of one promising high Z material, BiFeO3, is performed. http://archive.org/details/transportimaging109454760 US Navy (USN) author. Approved for public release; distribution is unlimited.