Bufo Melanostictus Research Articles

Updated Checklist of Amphibians of Pulau Jerejak, Penang, Peninsular Malaysia

Abstract A survey was carried out at Jerejak Island, Penang, Peninsular Malaysia from August 2008 to January 2011 to inventorise the amphibians contained therein. Specimens were either captured at night by hand or fishnets and then identified. The results showed that a total of 10 species of amphibians from 5 families were recorded during this survey, three of which have not previously been recorded in the Jerejak Island. The most common anuran species were the Common Asiatic Toad (Duttaphrynus melanostictus) and the Common Treefrog (Polypedates leucomystax) while the least common amphibian was the White-lipped Frog (Hylarana labialis).

Reproductive activity and advertisement calls of the Asian common toad Duttaphrynus melanostictus (Amphibia, Anura, Bufonidae) from Bach Ma National Park, Vietnam

We examined differences in reproductive activities and intraspecific variations in advertisement calls of Duttaphrynus melanostictus (Schneider, 1799) that lives in a tropical region of central Vietnam. The snout-vent length (SVL) of sexually mature males ranged from 50.2 to 70.3 mm, while that of females ranged from 65.2 to 97.3 mm. Histological analyses of testes revealed that sperm was not present throughout the year, indicating discontinuous reproductive activity for adult males. Adult females were captured year-round, but reproductive females were observed only during months of the auxiliary rainy season (mainly April to July), indicating that females reproduce seasonally. We also estimated levels of within-male variation of each call property and the influences of the ambient temperature, humidity, and the SVL of calling males on acoustic features. The call rate and pulse rate showed intermediate levels of variation, whereas the dominant frequency and call duration were the most stereotyped properties. One-way analyses of variance for six acoustic properties showed that the call rate, pulse rate, and rise time significantly differed (p < 0.05) among localities. Advertisement calls were a series of groups of 56 to 244 (145 ± 54) pulses with an average call duration of 26.722 s, an average pulse rate of 11.69 pulses/s, and an average dominant frequency of 1.293 kHz. The results of the multiple regressions for possible effects of temperature, humidity, and SVL on the six acoustic properties indicated that the dominant frequency, pulse rate, call duration, and rise time were positively significant. In Bach Ma National Park, when air temperature in the recording area decreased to <16.6°C, and advertisement calls of adult males virtually ceased in all three populations.

Bufalin inhibits migration and invasion in human hepatocellular carcinoma SK-Hep1 cells through the inhibitions of NF-kB and matrix metalloproteinase-2/-9-signaling pathways

Metastasis plays an important role in mortality of cancer patients. Migration and invasion are the major characteristics of tumor metastasis. The induction of matrix metalloproteinases (MMPs) such as MMP-2 and -9 are particularly important for the invasiveness of various cancer cells. Bufalin, a class of toxic steroids, was purified from the skin glands of Bufo gargarizans or Bufo melanostictus; it is known to inhibit proliferation of human cancer cells. In this study, we investigated the antiinvasive mechanisms of bufalin in the human hepatocellular cancer cell line SK-Hep1. Bufalin significantly reduced serum-induced cell invasion and migration. Furthermore, bufalin markedly inhibited MMP-2 and -9 activity, mRNA expression and protein levels in SK-Hep1 cells. Bufalin attenuated phosphoinisitide-3-kinase (PI3K) and phosphorylation of AKT which was associated with reduced levels of nuclear factor kappa B (NF-κB). Bufalin also suppressed protein levels of FAK and Rho A, VEGF, MEKK3, MKK7, and uPA and it diminished NF-κB translocation. Based on these observations, we propose that bufalin is acts as an antiinvasive agent by inhibiting MMP-2 and -9 and involving PI3K/AKT and NF-κB pathways. Bufalin is a potential therapeutic agent that may have efficacy in preventing the invasion and metastasis of malignant liver tumors.

Changes in Plasma Sex Steroids in Relation to Reproductive Behavior of the Toad, Bufo melanostictus (Schn.)

Steroid hormones play a critical role in the regulation of reproductive behavior of many amphibians. Changes in the plasma concentration of steroid hormones (testosterone, estradiol, and progesterone) in relation to sexual behavior of males and females were studied in a natural population of the toad (Bufo melanostictus) during the breeding season. Changes in plasma sex steroids were positively correlated with the sexual behavior of male and female toads. Plasma testosterone was low in calling males but reached a peak during amplexus before declining in the spawning and post-spawning males. Plasma estradiol and progesterone increased with the onset of sexual behavior in solitary females ready to breed, reaching a peak during spawning before declining in post-spawning females. The study reveals that B. melanostictus belongs to the type of ‘associated reproductive pattern’ in which plasma steroid hormone concentration is highly correlated with the sexual behavior of male and female toads.

Phylogenetic Relationship of Duttaphrynus melanostictus From India and China as Revealed from the Study of 12S and 16S mtDNA Genes

Phylogenetic Relationship of Duttaphrynus melanostictus From India and China as Revealed from the Study of 12S and 16S mtDNA Genes

Prey selection by tiger frog larvae (Hoplobatrachus chinensis) of two sympatric anuran species' tadpoles

We examined the prey selection and behavioral responses of tiger frog Hoplobatrachus chinensis larvae exposed to unpalatable and palatable sympatric prey tadpoles, Bufo melanostictus and Pelophylax nigromaculatus. We found that after a short exposure to the toxic toad tadpoles B. melanostictus, predators may learn to decrease going after unpalatable prey, subsequently it seems they may express short-term behavioral memory in order to avoid the toxic prey. In general, H. chinensis showed no preference for either any of the two prey species, which may be the result of P. nigromaculatus using behavioral performance and chemical defense as antipredatation strategies. These results facilitate further investigation of other aspects of the behavioral ecology of these three anuran species and hint at some potentially interesting possibilities of memory in choice of prey which may suggest further study.

A comparative study of hepatic mitochondrial oxygen consumption in four vertebrates by using Clark-type electrode

The present study was undertaken to establish a comparative account on hepatic mitochondrial oxygen consumption of Clarias gariepinus (fish), Bufo melanostictus (amphibian), Gallus gallus (bird) and Rattus norvegicus (mammal) and to correlate it with their specific metabolic rate (SMR). Mitochondrial oxygen consumption was measured with a Clarke-type electrode with succinate and pyruvate/malate as substrates. ADP was used to start state-III respiration. The results show that rats and chickens have higher oxygen consumption rate than that of fish and toads. Similarly, a species and substrate specific difference was also noticed in P/O (phosphate utilized per oxygen atom) ratio and respiratory control index. In case of rat, a significant negative correlation was noticed between P/O ratio and SMR with succinate as substrate. It is surmised that the observed difference in the mitochondrial respiration and P/O ratio in the above vertebrates is due to the difference in their metabolic activities.

Efficacy and safety of Huachansu combined with chemotherapy in advanced gastric cancer: A meta-analysis

BackgroundHuachansu, a Chinese medicine that comes from dried toad venom from the skin glands of Bufo bufo gargarizans Cantor or Bufo melanostictus Schneider, has been used in treatment of cancers. We conducted a meta-analysis to evaluate the efficacy and safety of Huachansu combined with chemotherapy for advanced gastric cancer. MethodsThe main efficacy measures were total response rates, quality of life and one-year survival rate. We also assessed the safety of treatments by evaluating the rate of occurrence of gastrointestinal side effects, leucocytopenia and neurotoxicity. The pooled relative ratio (RR) with 95% confidence interval (95% CI) was calculated to estimate the efficacy and safety of Huachansu combined with chemotherapy. ResultsFifteen randomized controlled trails were eligible. The result showed that Huachansu combined with chemotherapy was superior to simple chemotherapy treatment in increasing of total response rate [RR=1.28; 95% CI: (1.11, 1.18)] and Karnofsky score [RR=1.31; 95% CI: (1.18, 1.45)], and reducing gastrointestinal side effects [RR=0.71; 95% CI: (0.61, 0.82)], and leucocytopenia [RR=0.75; 95% CI: (0.64, 0.87)]; there was no significant difference on one-year survival rate (RR=1.25; 95% CI: 0.73, 2.14) between combination group and simple chemotherapy group. ConclusionCompared with chemotherapy control group, Huchansu combined with chemotherapy provide benefits for advanced gastric cancer on improving the response rate, increasing Karnofsky score, reducing leucocytopenia and major side effects such as gastrointestinal side effects caused by chemotherapy.

A new species of Duttaphrynus (Anura:Bufonidae) from Northeast India.

A new species of montane toad Duttaphrynus is described from Nagaland state of Northeast India. The new species is diagnosable based on following combination of characters: absence of preorbital, postorbital and orbitotympanic ridges, elongated and broad parotid gland, first finger longer than second and presence of a mid-dorsal line. The tympanum is hidden under a skin fold (in male) or absent (in female). The species is compared with its congers from India and Indo-China. We propose to consider Duttaphrynus wokhaensis as junior synonym of Duttaphrynus melanostictus.

Sex, organal and seasonal differences of Belminthofauna of toad, bufo melanostictus (Schneider, 1799)

A total of 75 toads (39 males and 36 females), Bufo melanostictus, collected from different areas of Dhaka city from May 2009 to April 2010 was examined for helminth parasites. The results showed that seven parasite species were collected, and prevalence of infection was 100%. The parasite fauna was comprised of Trematoda: 8% was the prevalence of Ganeo kumaonensis, 8% of Halipegus eccentricus; Nematoda: 100% of Oswaldocruzia indica, 68% of Neopharyngodon sp., 40% of Rhabdias bufonis; Acanthocephala: 28% of Pseudoacanthocephalus bufonis; and Pentastomida: 4% of Raillietiella indica. Serious damage was observed in intestine (53.94%) then rectum (24.55%), stomach (12.73%) and lungs (8.78%). The males had a higher intensity (77.92) than the females (44.58). Seasonally, the highest intensity of infection was found in the rainy season (81.21) and lowest during the winter (39.87). DOI: http://dx.doi.org/10.3329/bjz.v40i2.14308 Bangladesh J. Zool. 40(2): 155-164, 2012

Malformations and mortality in the Asian Common Toad induced by exposure to pleurolophocercous cercariae (Trematoda: Cryptogonimidae)

Malformations and increased mortality due to infection by the digenetic trematode, Riberioa ondatrae have been reported for many species of amphibians. Severe malformations have also been reported in the Common Hourglass Tree Frog, Polypedates cruciger induced by pleurolophocercous cercariae in Sri Lanka in addition to the changes in the behaviour, development and survival of the host. We exposed pre-limb bud stage tadpoles (Gosner stages 25–26) of the Asian Common Toad, Duttaphrynus melanostictus to the same pleurolophocercous type cercariae under laboratory conditions. Molecular and morphological identification showed that these cercariae belonged Acanthostomum burminis infecting freshwater snakes as definitive hosts. These cercariae induced malformations (27.8%) and reduced survival to metamorphosis (53.8%). The magnitude of the effects increased with the dose of cercariae. Types of malformations were mainly axial, such as scoliosis and kyphosis. Severe limb malformations such as extra or missing limbs as reported for amphibians exposed to R. ondatrae were not observed in the D. melanostictus. Same authors reported a higher percentage of malformations previously when P. cruciger was exposed to the cercariae A. burminis compared to D. melanostictus. However, tadpoles of D. melanostictus, which are smaller compared to those of P. cruciger, experienced higher mortality than P. cruciger tadpoles. Trematode induced malformations and mortality in amphibians are highly variable and depend on multiple factors such as host species differences such as resistance to infection and tolerance, life-history characteristics such as size at metamorphosis and length of the metamorphosis period, and other factors such as size of the amphibian at the time of trematode exposure.

Development of polymorphic microsatellite markers of the common toad, Duttaphrynus melanostictus useful for genetic studies

Asian common toad, Duttaphrynus melanostictus, is a widespread habitat generalist species of Indian-subcontinent and South-East Asia. Here we report, 14 new microsatellite markers (SSRs) for this toad, developed from a SSR-enriched genomic library, and validated using 44 unrelated samples of Asian toad and ~20 samples belonging to four congeners. Nine SSRs were highly polymorphic; amplified 6–22 alleles/marker with Ho and PIC ranging from 0.477 to 0.955 and 0.518 to 0.918, respectively, while one was found to be monomorphic. Of the remaining four SSRs, two (DMccmb02, DMccmb17) amplified only two apparently fixed alleles in all samples, while the other two (DMccmb22, DMccmb29) amplified 1–4alleles/sample; these four SSRs probably represent duplicated loci. Interestingly, all the 14 SSRs showed cross-species transferability to varying extent, with four SSRs showing 100 % transferability. It is hoped that these SSRs will greatly facilitate population genetic studies on the Asian toad species across their geographic distribution.

Observation of Frog Species in State University of Malang as a Preliminary Effort on Frog Conservation

Frog is an amphibian which widely distributed around the world. Indonesia houses 450 species which represent 11% of frog species in the world. In Java Island living 42 species of frogs and toads. Frogs can be used as an environment indicator due to the presence of frog in a particular place indicates that the place is stay natural and unpolluted. State University of Malang Campus #1 which is located in the heart of Malang District has been developing rapidly, currently. Thus, it requires for the construction of new various facilities to support it widen activities. Extensive construction can be destructive even damaging to the habitat of frog, which potentially threat the frog life, if it does not take a consideration on the environmental impact, carefully. This study aimed to identify the species of frog which is survives in State University of Malang with consideration of the frog species found in 1995. Species identification conducted by observing the morphologicalcharacter. This study found four species with three species which are survive and as found in 1995; those are Duttaphrynus melanostictus, Polypedates leucomystax, and Kaloula baleta; and one new species that is Rana chalconota. This study also reveals there are four species which are extinct; those are Fejervarya cancrivora, Fejervarya limnocharis, Ingerophrynus biporcatus, and Occidoziga lima. This situation shows the decreasing amount of species from 7 to 4 within 17 years. This result indicates that there is a serious environmental degradation which caused the losing of frog habitats. Further research is needed to study the ecological condition changing in order to save the frog species.

Observation of Frog Species in State University of Malang as a Preliminary Effort on Frog Conservation

Frog is an amphibian which widely distributed around the world. Indonesia houses 450 species which represent 11% of frog species in the world. In Java Island living 42 species of frogs and toads. Frogs can be used as an environment indicator due to the presence of frog in a particular place indicates that the place is stay natural and unpolluted. State University of Malang Campus #1 which is located in the heart of Malang District has been developing rapidly, currently. Thus, it requires for the construction of new various facilities to support it widen activities. Extensive construction can be destructive even damaging to the habitat of frog, which potentially threat the frog life, if it does not take a consideration on the environmental impact, carefully. This study aimed to identify the species of frog which is survives in State University of Malang with consideration of the frog species found in 1995. Species identification conducted by observing the morphologicalcharacter. This study found four species with three species which are survive and as found in 1995; those are Duttaphrynus melanostictus, Polypedates leucomystax, and Kaloula baleta; and one new species that is Rana chalconota. This study also reveals there are four species which are extinct; those are Fejervarya cancrivora, Fejervarya limnocharis, Ingerophrynus biporcatus, and Occidoziga lima. This situation shows the decreasing amount of species from 7 to 4 within 17 years. This result indicates that there is a serious environmental degradation which caused the losing of frog habitats. Further research is needed to study the ecological condition changing in order to save the frog species.

Quality Assessment of Serially Ultradiluted and Agitated Drug Digitalis purpurea by Emission Spectroscopy and Clinical Analysis of Its Effect on the Heart Rate of Indian Bufo melanostictus.

The investigation of ultradiluted (homeopathic) drugs is extremely interesting and challenging, and from that point of view this study shows novelty. A study of in vivo changes in heart rate of the Indian Bufo melanostictus caused by commercially available serially ultra-diluted and agitated extract of Digitalis purpurea has been tried in order to understand their pharmacological role. RR interval (of ECG) was compared after intraperitoneal administration of serially diluted and agitated Digitalis purpurea extract, diluent rectified spirit, and Digoxin in anesthetized animals. The study revealed statistically significant changes in the heart rate after application of these drugs except in case of Digoxin and the 200th serial dilution of Digitalis purpurea. The duration of RR intervals after application of the drugs was corroborative of the effect of Digoxin and Digitalis purpurea extract up to 30th dilution. Emission spectra were obtained for the experimental ultra-diluted Digitalis purpurea extract and Digoxin to identify and characterize them. The observed RR pattern and emission spectra show an association. The quality assessment of the commercial ultra-diluted organic drugs obtained from natural products may be initiated by monitoring in vivo studies on animal models.

Chlorpyrifos-induced Histopathological Alterations in Duttaphrynus melanostictus (Schneider 1799) Tadpoles: Evidence from Empirical Trials

This paper reports the histopathological responses of the gill, liver and tail muscle tissues in tadpoles of the Asian Common Toad Duttaphrynus melanostictus (Schneider, 1799) exposed to chlorpyrifos a common organophosphorus pesticide. Tadpoles of Gosner stages 24-26 were continuously exposed to low, mid and high (500, 1000 and 1500 µgl–1) concentrations of chlorpyrifos for two weeks. Histological alterations in the tissues of the surviving larvae were microscopically examined both at the end of the exposure period and after a week following the final exposure. Several histological alterations were noted in the gills, liver and tail muscles of the larvae exposed to 1500 µgl–1 of chlorpyrifos. The gills of exposed larvae showed architectural distortion resulting from reduced primary and secondary gill lamellae and blood vessels, and alterations in the gill epithelium. In the liver sinusoidal congestion and dilation, cytoplasmic vacuolation and changes in hepatocytes such as hyperchromatic nuclei and nuclear fragmentation were observed. The tail muscle tissue suffered from severe atrophy and myotomal disintegration. Although histological alterations in the gill and liver tissues were noted only at the high concentration, changes in the muscle tissue i.e. muscle degeneration and atrophy, were apparent at both low and mid concentrations. The degree of damage in surviving larvae in a week following the final exposure was lower than that observed during the exposure, probably indicating recovery or resistance. The findings of the present study emphasize the need to investigate possible sublethal damage induced by pesticides in amphibians inhabiting agricultural habitats.

Mating Pattern, Spawning Behavior, and Sexual Size Dimorphism in the Tropical ToadBufo melanostictus(Schn.)

Pattern of mating, spawning behavior, and sexual size dimorphism were studied in the Indian Common Toad, Bufo melanostictus. Male and female B. melanostictus are sexually dimorphic in size. Adult males are significantly smaller than adult females, and the ratio of body size of females to males was 1:19. For breeding, the majority of males and females segregated on the basis of their size; larger males paired with larger females leaving smaller males to pair with smaller females. A positive correlation between the body sizes of mating partners provided evidence of size-assortative mating. Successful males were larger in length than their unsuccessful competitors. Operational sex ratio was skewed in the favor of males. A stereotypical spawning behavior was observed: At the initiation of spawning by the female, the male juxtaposed his cloaca with that of the female's followed by a few seconds of rigorous toe movements. Spawning was completed within 2–5 h. Subsequently, the female exhibited pseudo-spawning behavior once or twice. In the absence of more eggs, the male quickly released his clasp, and the pair separated.

Advertisement Call Variability in the Black-spined Toad &lt;I&gt;Bufo melanostictus&lt;/I&gt; (Anura: Bufonidae) during the Breeding Season in Lishui, Zhejiang, China

Advertisement Call Variability in the Black-spined Toad &lt;I&gt;Bufo melanostictus&lt;/I&gt; (Anura: Bufonidae) during the Breeding Season in Lishui, Zhejiang, China

166. Transformation of Bidder's Organ into Ovary in Tadpoles of Bufo melanostictus Chronically Exposed to Endocrine Disrupters.

166. Transformation of Bidder's Organ into Ovary in Tadpoles of Bufo melanostictus Chronically Exposed to Endocrine Disrupters.

Dose dependent interactions between estradiol‐17β and triiodothyronine with reference to different physiological and biochemical parameters on liver tissue of common Indian Toad (Bufo melanostictus)

To study the dose dependent interaction between two chemically differentand functionally distinct hormones i.e. estradiol‐17β and tri‐iodothyronine(T3), six different animal groups of common Indian toad (Bufo melanostictus)were injected intramuscularly each with single dose of individual treatment[Estradiol‐17β (E‐40 and E‐80); T3 (T‐20 and T‐40), and combination dose (E‐40+T‐20 and E‐80+T‐40)]. The data clearly demonstrated the interaction between these two hormones with respect to different physiochemical parameters in the liver tissue, such as alkaline phosphates activity, oxygen consumption rate, lipid, cholesterol, RNA, DNA, protein, free amino acid, totalsugar and ascorbic acid content with respect to different days after treatment(0, 1, 5 and 10) as well as different doses of hormonal treatments, individuallyand in combination. The comparative study indicated the dose dependenthormone‐hormone interaction between estradiol‐17β and T3 hormone with respect to synergistic effect on oxygen consumption rate, fluctuation in lipid and cholesterol content, antagonistic effect of T3 on activation of the transcriptional apparatus to synthesize RNA, free amino acid metabolism, antagonistic effect on protein metabolism suggesting the protein catabolic or anabolic nature of T3, the permissive effect of estradiol‐17β upon T3 on sugar content in liver tissue of toad.