A cryptic Bornean torrent frog of the genus Meristogenys, which is divergent genetically and morphologically from all known congeners, is described from mountain streams of western Sarawak, East Malaysia (Borneo). The species occurs sympatrically with the type species of the genus, M. jerboa, but apparently differs from it in adult coloration and larval morphology, such as keratodont formulae and glands in tail fins. Females of the new species possess much larger and fewer eggs than in sympatric M. jerboa, suggesting significantly different reproductive traits between these species. A key to larvae of known species of the genus is provided.
We describe a new species of cascade frog of the genus Rana, from west Malaysia. Rana monjerai, new species is a medium-sized frog of the subgenus Odorrana (SVL of males, 38-43 mm; of one female, 75 mm), and is distinguished from all other members of this subgenus by the combination of: white lip stripe, dorsolateral fold, full web on the fourth toe, vomerine teeth, gular vocal pouch and relatively large tympanum in males, no dorsal marking, no clear light spots on rear of thigh, first finger subequal to second, finely tuberculated dorsum, and unpigmented ova. The significance of finding this species from peninsular Malaysia is discussed.
We describe a new species of torrent-dwelling ranid frog of the genus Meristogenys from the Crocker Range, western Sabah, northern Borneo. The new species, Meristogenys maryatiae, differs from congeners by the combination of: small body, males 31-37 mm and females 65-66 mm in snout-vent length; head narrower than long; eyes moderate, diameter subequal to snout; iris unicolored; legs long; ventral surface of tibia without heavy pigmentation; rear of thigh blotched dark brown and cream; toes fully webbed; outer metatarsal tubercle present; larval dental formula 7(4-7)/6(1).
Infection with Chloromyxum careni Mutschmann, 1999 was found in the Asian horned frog Megophrys nasuta from Malaysia and Indonesia. Kidney was the only organ infected. Coelozoic plasmodia up to 300 μm were localized in Bowman's space, embracing the glomerulus from all sides, or rarely in lumina of renal tubules. Plasmodia are polysporic, containing disporic pansporoblasts. Myxospores observed by light microscopy are colorless, variable in shape and size, measuring 6.0-8.5 × 5.0-6.5 μm, composed of two symmetrical valves joined by a meridian suture, containing four pyriform polar capsules 3.0-4.0 × 2.5-3.0 μm and a single sporoplasm. Each valve possesses 14-24 (median 21) fine longitudinal ridges clearly visible only in scanning electron microscopy. Rarely, atypical spores with a markedly pointed posterior pole and only 6-10 surface ridges are present in plasmodia together with typical spores. Both small subunit (SSU) and large subunit (LSU) rRNA gene sequences possess extremely long GU-rich inserts. In all SSU and LSU rDNA-based phylogenetic analyses, C. careni clustered as a distinct basal branch to the Myxobolus+Myxidium lieberkuehni clade, out of the marine Chloromyxum clade containing Chloromyxum leydigi, the type species of the genus. These morphological and phylogenetic data suggest erection of a new genus for the C. careni lineage, but we conservatively treat it as a Chloromyxum sensu lato until more information is available.
Boesenbergia rotunda, traditionally used to relieve stomach, abdomen, joint, muscle, and rheumatic pain was also reported for its antinociceptive effect on a mouse model. However, the possible pain relief effect of Boesenbergia rotunda ethanolic extract (BREE) via the inhibition to the neural pain pathway remains to be elucidated. This study investigated the inhibitory effect of BREE on compound action potentials (CAPs) and the possible involvement of the opioid receptors. The changes in the CAPs amplitudes of the frog’s sciatic nerves were evaluated following the exposure to three different dosages of BREE (1, 3 and 10 mg/ml and morphine (3 mg/ml). In another set of experiment, the nerves were pretreated with a non-selective opioid receptor antagonist, naloxone (0.1 mg/ml), before exposing the nerve to BREE (1 mg/ml) to investigate the involvement of opioid receptors in the CAPs inhibitory mechanism. The outcome showed a reduction in the CAPs amplitudes when treated with BREE (1, 3 and 10 mg/ml) whereby the effect was reversible. The CAPs inhibition by BREE was absent when the opioid receptors were blocked. Taken together, these findings suggest that BREE-induced CAPs amplitude reduction involves the activation of opioid receptors.
A survey was conducted to catalogue the diversity of anurans in District IV of the Muda Agriculture Development Authority Scheme (MADA) in Kedah Darul Aman, Malaysia, from July 1996 to January 1997. Eight species of anurans from three families were present in the study area. Of these, the Common Grass Frog (Fejevarya limnocharis) was the most abundant, followed by Mangrove Frog (Fejevarya cancrivora), Long-legged Frog (Hylarana macrodactyla), and Common Toad (Duttaphrynus melanostictus). Puddle Frog (Occidozyga lima), Taiwanese Giant Frog (Hoplobatrachus rugulosus), and Banded Bullfrog (Kaluola pulchra) were rare during the sampling period, and only one Paddy Frog (Hylarana erythraea) was captured. A simple identification key for the anurans of this area is included for use by scientists and laymen alike.
Bukit Perangin Forest Reserve, Kedah Malaysia is a popular forest park well known for its clear rivers and streams, interesting sedimentary rock formations and a rich biodiversity in flora and fauna. However to date no report on the amphibians and reptiles of this forest has ever been published. Therefore, from January 2009 to May 2010, herpetofauna surveys were conducted at Bukit Wang Recreational Forest, which forms a part of the Bukit Perangin Forest Reserve. The results of the survey showed that a total of 15 species of amphibians belonging to 6 families and 15 species of reptiles
belonging to 7 families were found in the study area. These constitute the first records for the amphibians and reptiles in this forest. The herpetofauna checklist showed that the most abundant amphibians were from the family Ranidae while the most abundant reptiles were from the family Gekkonidae.
Rakaman panggilan katak Borneo daripada genus Hylarana telah dijalankan di tujuh lokasi kajian di Sarawak, Malaysia. Sebanyak 12 individu daripada lima spesies katak Borneo telah berjaya dirakam dan dianalisis. Daripada 38 pembolehubah panggilan, hanya 27 daripadanya dapat menerangkan ciri-ciri panggilan daripada katak Hylarana Borneo. Saiz katak jantan mempengaruhi secara signifikan tenaga panggilan antara 90:50% amplitud puncak, tempoh nadi di antara 50%, nisbah tempoh panggilan untuk mencapai setengah modulasi frekuensi dan kadar ulangan not. Saiz katak Hylarana Borneo sedikit mempengaruhi frekuensi asas dan dominan panggilan. Di samping itu, taburan pembolehubah panggilan katak dan dendogram Euclid menyokong identiti taksonomi bagi setiap spesies katak Borneo Hylarana.
The tiger frog Hoplobatrachus rugulosus (Wiegmann 1834) is a large robust dicroglossid frog widely distributed in southern China, Malaysia, Myanmar, Vietnam and Thailand. The escaped bred tiger frog introduced from Thailand hybridized with Chinese native population may have affected the genetic diversity of local Chinese tiger frogs. However, previous microsatellite loci of this species do not offer enough information to construct the genetic map. Here, we reported 33 new microsatellite loci from transcriptome sequencing for H. rugulosus. Alleles ranged between 1 and 10 per locus and only one locus (HRT001) was monomorphic. The polymorphic information content, observed and expected heterozygosity were 0-0.794, 0-0.969 and 0-0.831, respectively. None of the loci was observed in linkage disequilibrium and two loci (HRT023 and HRT068) deviated from Hardy-Weinberg equilibrium after Bonferroni correction for multiple tests. These transcriptome-derived microsatellite markers will be usedto study the genetic divergence and construct the genetic map in H. rugulosus.
he Chinese tiger frog Hoplobatrachus rugulosus is widely distributed in southern China, Malaysia, Myanmar, Thailand, and Vietnam. It is listed in Appendix II of CITES as the only Class II nationally-protected frog in China. The bred tiger frog known as the Thailand tiger frog, is also identified as H. rugulosus. Our analysis of the Cyt b gene showed high genetic divergence (13.8%) between wild and bred samples of tiger frog. Unexpected genetic divergence of the complete mt genome (14.0%) was also observed between wild and bred samples of tiger frog. Yet, the nuclear genes (NCX1, Rag1, Rhod, Tyr) showed little divergence between them. Despite this and their very similar morphology, the features of the mitochondrial genome including genetic divergence of other genes, different three-dimensional structures of ND5 proteins, and gene rearrangements indicate that H. rugulosus may be a cryptic species complex. Using Bayesian inference, maximum likelihood, and maximum parsimony analyses, Hoplobatrachus was resolved as a sister clade to Euphlyctis, and H. rugulosus (BT) as a sister clade to H. rugulosus (WT). We suggest that we should prevent Thailand tiger frogs (bred type) from escaping into wild environments lest they produce hybrids with Chinese tiger frogs (wild type).
Previously, only one species of torrent frog (Amolops larutensis) was thought to occur throughout Peninsular Malaysia. However, genomic work has demonstrated that populations from eastern Peninsular Malaysia form two separate lineages that are genetically distinct from A. larutensis that is now restricted to the western half of Peninsular Malaysia. This study demonstrates that all three lineages can be morphologically distinguished from each other, thereby providing additional support for the recognition of the eastern lineages as two distinct species. These lineages are described herein as Amolops gerutu sp. nov. from the eastern states of Kelantan, Terengganu, and Pahang, and A. australis sp. nov. from the southern-most state of Johor. In general, these two new species form a clade that is sister to A. larutensis and can be readily distinguished from it by having: (1) considerably denser and more pronounced dorsal tubercles, and (2) the posterodorsal surface of thighs having dense, dark stippling as opposed to broad vermiculations. Although differences in other morphometric characters were detected, their utility as diagnostic characters should be applied with caution due to the large intraspecific variation that overlaps among different species in many of the characters we measured. As such, we advocate for the use of tuberculation and pattern of the posterodorsal portion of the thighs as primary diagnostic characters. These characters can readily distinguish A. larutensis from the two new species. To differentiate A. australis sp. nov. from A. gerutu sp. nov. and A. larutensis, body size can be a good diagnostic character as A. australis sp. nov. is significantly smaller in both males (mean = 31.04 ± 1.59 mm) and females (mean = 46.48 ± 3.2 mm). Additionally, we show a strong positive correlation between body size and elevation, with populations from montane forests (>900 m asl) being considerably larger than populations at lower elevations.
The present study was conducted to elucidate the genetic divergence and the phylogenetic relationships in the F. limnocharis complex from Bangladesh and other Asian countries such as Sri Lanka, Thailand, Malaysia, Taiwan and Japan by allozyme analyses. We used a total of 95 frogs of the F. limnocharis complex from these countries and F. cancrivora from the Philippines as an outgroup. Based on body size, the F. limnocharis complex from Bangladesh was divided into three distinct groups: large, medium and small types. Allozyme analyses were carried out with 28 loci encoding 20 enzymes and two blood proteins by horizontal starch-gel electrophoresis. When genetic distance was calculated, distinct divergence was found among the three types: mean genetic distance was 0.782 between the small and medium types, 1.458 between the large and medium types, and 1.520 between the large and small types. Phylogenetic trees based on genetic distance showed that all populations of Bangladesh small type strongly formed a cluster and were found to be most closely related to the Sri Lanka population; that all populations of Bangladesh large type formed a very strong cluster and were grouped with several populations from Thailand, Malaysia, Japan, and Taiwan; and that the medium type was segregated from all other groups. This may imply that each of the three types is a different species, and that the medium type is possibly an undescribed taxon.
Glutathione-S-Transferase (GST) and metallothionein are important biomarker endpoints in studying the effect of Cd exposure. The purpose of this research was to study the correlation between hepatic GST and metallothionein with hepatic Cd in wild Fejervarya limnocharis exposed to environmental Cd. Results showed that frogs from contaminated sites had significantly higher hepatic metallothionein (3.58 mg/kg wet weight) and GST activity (0.259 μmol/min/mg total protein) than those from the reference site (2.36 mg/kg wet weight and 0.157 μmol/min/mg total protein respectively). There was a significantly positive correlation between hepatic Cd and GST activity (r = 0.802, p = 0.009) but not between hepatic Cd and metallothionein (r = 0.548, p = 0.139). The results concluded that while frogs from the contaminated site had higher GST and metallothionein, only GST showed significant positive correlation with hepatic Cd levels, indicating that hepatic GST activity may be used as a biomarker endpoint.
A survey was carried out at Gunung Panti Forest Reserve, Johor from 3-7 August 2006, 2-5 June 2008, and 28-31 July 2008 to inventory the herpetofauna therein. An updated checklist for the area which incorporates findings from previous studies is provided. In total, 37 species of frogs, 1 turtle, 27 lizards, and 11 snakes have been recorded from Gunung Panti Forest Reserve, Johor.
Recordings of frog calls from the genus Hylarana were conducted at seven study sites in Sarawak, Malaysia. The results showed that each species differs in terms of call characteristics, in which a high number of pulsed note belongs to H. baramica, while high repetition note belongs to the sibling species, H. glandulosa. Higher pitched and number of amplitude spectrum, however, belong to H. signata. Pulsed note and note repetition differed significantly among species, except for spectrum amplitude. Advertisement calls for each species were described in detail. This study provide a baseline data on advertisement call characteristics of frogs from the genus Hylarana for future studies of these frogs particularly and the Oriental frogs generally.
The sound produced by the anurans including toads can be a survival strategy for the selection of fittest males by their conspecific females and a form of effective prezygotic mechanism. Most of the published works on anuran sound production were on true frogs or ranids, yet little is known about sound characteristics of toads. Here we describe the properties of mating calls from six selected toad species from six study sites in Sarawak. Males' mating calls were recorded and analyzed using Sound Ruler Acoustic Analysis ver 0.9.6.0. Call characteristic were subsequently compared for toad species differentiation. The results showed that each species differ in call characteristics, with pulsating note, rate of note repetition and pitch being the most apparent characters. The highest number of pulse notes belongs to Duttaphrynus melanostictus while Ansonia spinulifer exhibited highest pitch and rate of note repetition. These species' differences in mating call characteristics provide an additional method for anuran species classification besides morphological and molecular DNA data. It is also a very useful guide during field survey as the sound is audible and identification can be made without the need of capturing the toads.
Mesocoelium malayanum sp. n. is described from the frog Rana macrodon, in Malaysia. Elongate body, broader anteriorly, measuring 1.900 (1.679-2.070) mm long by 0.404 (0.380-0.437) wide, tegument aspinose oral sucker 0.212 (0.200-0.228) by 0.202 (9.191-0.205), acetabulum 0.141 (0.132-0.150) by 0.139 (0.123-0.146), prepharynx present, oesophagus 0.115 (0.096-0.137), caeca reaching posterior 1/3 of body, anterior testis 0.097 (0.087-0.110) by 0.091 (0.087-0.100) dorsal to acetabulum, posterior testis 0.094 (0.087-0.101) by 0.092 (0.091-0.100), cirrus pouch 0.121 (0.111-0.130) by 0.047 (0.041-0.055), genital pore at left of midline of oesophagus just anterior to intestinal bifurcation, ovary 0.110 (0.091-0.127) by 0.089 (0.085-0.096) on left of body and posterior to acetabulum, vitelline glands with single follicles extending from intestinal bifurcation to ends of caeca, excretory vesicle I-shaped and eggs 0.040 (0.037-0.046) by 0.023 (0.022-0.024). Although morphologically related to M. maroccanum and M. meggitti, M. malayanum is considered to be a new species.
The genus Rana, notably diversified in Oriental regions from China to Southeast Asia, includes a group of cascade frogs assigned to subgenera Odorrana and Eburana. Among them, R. ishikawae and the R. narina complex represent the northernmost members occurring from Taiwan to the Ryukyu Archipelago of Japan. Relationships of these frogs with the continental members, as well as the history of their invasions to islands, have been unclear. The taxonomic status of Odorrana and related genera varies among authors and no phylogenetic reassessment has been done. Using partial sequences of mitochondrial 12S and 16S rRNA genes, we estimated phylogenetic relationships among 17 species of the section Hylarana including Odorrana and Eburana, and related species from the Ryukyus, Taiwan, China, Thailand, Malaysia, and Indonesia. We estimate that (1) Odorrana is monophyletic and encompasses species of Eburana and R. hosii, which is now placed in Chalcorana, (2) the ancestor of R. ishikawae separated from other Rana in the middle to late Miocene prior to its entry to the Ryukyu Archipelago, (3) the ancestor of the R. narina complex later diversified in continental Asia, and invaded the Ryukyu Archipelago through Taiwan, (4) the R. narina complex attained its current distribution within the Ryukyus through niche segregations, and (5) vicariance of R. hosii between Malay Peninsula and Borneo occurred much later than the divergence events in the R. narina complex. Current subgeneric classification of Rana, at least of Southeast Asian members, requires full reassessment in the light of phylogenetic relationships.
Crude glycoside extracts from the plant, Sarcolobus globosus, were tested on the rat phrenic nerve-diaphragm, chick biventer cervicis and frog rectus abdominis preparations. Nerve-stimulated twitches were inhibited by the extract. The muscle paralysis was not similar to that by curare-like blockers as it was not reversed by neostigmine or by a tetanus. Although contractures to acetylcholine or carbachol were not affected by 0.6 mg/ml of the extract, higher concentration of the extracts (3 mg/ml) depressed the log dose-response curve of acetylcholine and carbachol. The results suggest that the neuromuscular blocking effect of the extracts is either dose-dependent or due to a mixture of toxins with presynaptic or postsynaptic actions.