Rana Tigrina Research Articles

Significance of Chronic Synthetic Pyrethroid Fenvalerate Administration on Carbohydrate metabolism in the Muscle, Brain and Intestine of Rana tigrina (Indian Bull frog)

Significance of Chronic Synthetic Pyrethroid Fenvalerate Administration on Carbohydrate metabolism in the Muscle, Brain and Intestine of Rana tigrina (Indian Bull frog)

Cardiotoxic Effect of Aqueous and Alcoholic Crude Extract of some Indigenous Plants on Rana Tigrina (Hoplobatrachus Tigerinus)

Cardiotoxic Effect of Aqueous and Alcoholic Crude Extract of some Indigenous Plants on Rana Tigrina (Hoplobatrachus Tigerinus)

Effect of Endosulfan on Intestine and Pancreas of frog Rana tigrina

Now a day's use of pesticides is the matter of great discussion for environmentalist to gain more relevant information on tolerance of organism to the pesticides. Rana tigrina were exposed to sub lethal concentration of Endosulfan (0.073 ml/lit) for 7 days regularly. The globlet cells of intestine was found to be swollen, enlargement of cells, cell shrinkage, damage of cell membrane, vacuoles formation and pancreas exocrine secreatory acini cells was found to be damaged and connective tissue was loosely bounded also observed on exposure to Endosulfan.

Changes in the Microbial Communities of Tiger Frog (Rana tigrina) Meat during Refrigerated Storage

Microbial activity is the major cause of the spoilage of aquatic meat products during storage. This study investigated the changes of the microbial compositions of the tiger frog (Rana tigrina) meat stored aerobically at 4°C for 12 days using 16S rRNA amplicon high-throughput sequencing analysis. The microbial diversity and species richness of the frog meat were abundant at the initial phase of storage but decreased substantially with prolongation of the storage time. Proteobacteria was the prevalent phylum identified from the frog meat, with a relative abundance of 40.29% at day 0 increasing to 96.77% at day 6 and 95.41% at day 12. At the genus level, Shewanella, Pseudomonas, and Acinetobacter were the three dominant genera in the spoiled samples and contributed to frog meat spoilage. Their proportions were 41.67, 28.48, and 5.94% at day 6 and 29.94, 23.48, and 18.44% at day 12, respectively. The present study is conducive to understanding the pattern and process of frog meat spoilage during refrigeration and could be used to develop efficient control measures to mitigate the predominant psychrotrophic spoilers in aerobically stored frog meat.

Phylogenetic position of Tremiorchis ranarum Mehra and Negi, 1926 (Trematoda: Plagiorchiidae) with remark on this monotypic genus.

Tremiorchis is a monotypic genus of digenetic trematode (Plagiorchiidae: Plagiorchiinae), infecting the frogs Rana tigrina (Hoplobatrachus tigerinus) and R. cyanophlyctis (Euphlyctis cyanophlyctis). Metacercaria use to infect Rana tigrina (Hoplobatrachus tigerinus) and R. cyanophlyctis (Euphlyctis cyanophlyctis) as intermediate hosts, while the cercaria stage found from apple snail, Pila virens. Adults of T. ranarum harbor mature frogs of H. tigerinus and E. cyanophlyctis. Besides the frequent infection of Tremiorchis, no DNA sequence data are currently available for this monotypic genus. The present communication, deals with the sequence data for nuclear ribosomal genes, 18S, small internal transcribed spacers (ITS1-5.8S-ITS2) and 28S to molecularly characterize T. ranarum. Besides this, phylogenetic relationship among the members of the Plagiorchiida is also discussed in detail. An attempt has also been made to provide detailed molecular affinities of T. ranarum with other trematode genera.

Acute toxic effects on hematological and histological features in adult frogs, Rana tigrina exposed to lambda-cyhalothrin pesticide

Acute toxic effects on hematological and histological features in adult frogs, Rana tigrina exposed to lambda-cyhalothrin pesticide

Acute toxic effects on hematological and histological features in adult frogs, Rana tigrina exposed to lambda-cyhalothrin pesticide

Acute toxic effects on hematological and histological features in adult frogs, Rana tigrina exposed to lambda-cyhalothrin pesticide

Transformation of Rana tigrina during Progressive Metamorphosis

We determined the kinetics of metamorphosis, apoptosis, and tail regression in Rana tigrina. Acid phosphatase activity (µMole Pi.hr-1.tail-1) in the growing and regressing tail attended six to thirty fold increase respectively. However total activity in the trunk was decreased through progressive growing stages of metamorphosis. Total protein content in the trunk of tadpoles at climax stage (XXI) was decrease (35%) from 2.6mg/ml to 1.7mg/ml. The tail of tadpole tissue has shown a two fold increase in total Ribonucleic Acid (RNA) content from stage III to stage XVIII. But there was again decrease in total RNA content at climax stage (stage XXI). This might be possible due to decreased protein synthetic status. When the experiment was performed in trunk homogenate the amount of total carbohydrate (mg/ml) was slightly increased from 37mg/ml to 38.6mg/ml. this might be due to increase in the activity of α-amylase enzymes in the viscera of developing tadpole when it reached the climax stage.

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This study focused on the quantitative determination of Cu metal in various physical parts of frogs by using the Atomic Absorption Spectrometry technique). Cu was measured (2.72±0.77 µg/g) in kidney of Rana tigrina captured from industrial site and in R. tigrina captured from non-industrial site (28.10±27.02 µg/g). Similarly, 27.56±9.92 µg/g Cu was discovered in liver of R. tigrina captured from industrial site and 6.56±3.06 µg/g in R. tigrina captured from non-industrial site. From the results of the present study, it may be concluded that frogs are good bio-indicator species to estimate the level of heavy metals pollution of various water sources in some countries.   Key words: Copper, bio-indicator, atomic absorption spectrometry (AAS), aquatic habitat.

Tiger frog virus ORF104R interacts with cellular VDAC2 to inhibit cell apoptosis

Ranaviruses belong to the family Iridoviridae, and have become a serious threat to both farmed and natural populations of fish and amphibians. Previous reports showed that ranaviruses could encode viral Bcl-2 family-like proteins (vBcl-2), which play a critical role in the regulation of cell apoptosis. However, the mechanism of ranaviruses vBcl-2 interactions with host protein in mediating apoptosis remains unknown. Tiger frog virus (TFV) belonging to the genus Ranavirus has been isolated from infected tadpoles of Rana tigrina rugulosa, and it causes a high mortality rate among tiger frog tadpoles cultured in southern China. This study elucidated the molecular mechanism underlying the interaction of TFV ORF104R with the VDAC2 protein to regulate cell apoptosis. TFV ORF104R is highly similar to other ranaviruses vBcl-2 and host Mcl-1 proteins, indicating that TFV ORF104R is a postulate vBcl-2 protein. Transcription and protein expression levels showed that TFV orf104r was a late viral gene. Western blot results suggested that TFV ORF104R was a viral structural protein. Subcellular localization analysis indicated that TFV ORF104R was predominantly colocalized with the mitochondria. Overexpressed TFV ORF104R could suppress the release of cytochrome C and the activities of caspase-9 and caspase-3. These results indicated that TFV ORF104R might play an important role in anti-apoptosis. Furthermore, the interaction between TFV ORF104R and VDAC2 was detected by co-immunoprecipitation in vitro. The above observations suggest that the molecular mechanism of TFV-regulated anti-apoptosis is through the interaction of TFV ORF104R with the VDAC2 protein. Our study provided a mechanistic basis for the ranaviruses vBcl-2-mediated inhibition of apoptosis and improved the understanding on how TFV subverts host defense mechanisms in vivo.

Gold in male reproductive tract of frog (Rana tigrina): a chronobiological study

Various elements present in the male reproductive tract and in semen of man and of different animals were reported (Skandhan and Abraham 1984; Skandhan and Pandya 1986; Skandhan et al. 2009). Prese...

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Some creatures are very sensitive to water borne pollutants and can be used as useful bio-indicators of water pollutions. The present study used these bio-indicators to determine the status regarding health of freshwater habitats. The study was carried out to quantify Zn in kidney and liver of Rana tigrina from different aquatic habitats (canal, fish pond and sewage water) by atomic absorption spectrometry (AAS). Liver and kidney tissues of R. tigrina from sewage habitat showed elevated levels of Zn compared to those collected from canal and fish pond water habitats. Hence, it may be concluded that R. tigrina is one of the good bio-indicator species for heavy metals. It can be used to assess the heavy metal load and pollution of aquatic ecosystems. Key words: Zinc, Rana tigrina, atomic absorption spectrometry (AAS), aquatic habitat.

Caveolae Restrict Tiger Frog Virus Release in HepG2 cells and Caveolae-Associated Proteins Incorporated into Virus Particles.

Caveolae are flask-shaped invaginations of the plasma membrane. Caveolae play important roles in the process of viruses entry into host cells, but the roles of caveolae at the late stage of virus infection were not completely understood. Tiger frog virus (TFV) has been isolated from the diseased tadpoles of the frog, Rana tigrina rugulosa, and causes high mortality of tiger frog tadpoles cultured in Southern China. In the present study, the roles of caveolae at the late stage of TFV infection were investigated. We showed that TFV virions were localized with the caveolae at the late stage of infection in HepG2 cells. Disruption of caveolae by methyl-β-cyclodextrin/nystatin or knockdown of caveolin-1 significantly increase the release of TFV. Moreover, the interaction between caveolin-1 and TFV major capsid protein was detected by co-immunoprecipitation. Those results suggested that caveolae restricted TFV release from the HepG2 cells. Caveolae-associated proteins (caveolin-1, caveolin-2, cavin-1, and cavin-2) were selectively incorporated into TFV virions. Different combinations of proteolytic and/or detergent treatments with virions showed that caveolae-associated proteins were located in viral capsid of TFV virons. Taken together, caveolae might be a restriction factor that affects virus release and caveolae-associated proteins were incorporated in TFV virions.

Tocolytic plant Tectona grandis Linn. extended study on other systemic effect.

Introduction: Tectona grandis Linn. (T.G) is one of the well-known Indian herbs. In Ayurveda, T.G stem extract has tocolytic effect. Our previous study conformed the tocolytic activity of the T.G stem extract on rat uterine contraction. Conventional tocolytic drugs are known to have cardiovascular, skeletal muscle toxicity. This study is undertaken to observe possible adverse effects of T.G stem extract especially on cardiovascular and musculoskeletal system. Materials and Methods: Wistar rats (Rattus norvegicus) and frogs (Rana tigrina) were used for this study. Animals were divided into six groups. Each group contains six animals. To study the effect on the blood vessels and skeletal muscles, frogs were used and for direct cardiac effects, Wistar rats were used. The results were compared with standard drugs such as sodium nitrate (2%), nifedipine, and magnesium sulfate. Results: After analysis by t-test, it was observed that T.G causes vasodilatation same as that of the sodium nitrate, without causing any cardiac toxicity seen with nifedipine and no neuromuscular blockade as seen with magnesium sulfate. Conclusion: T.G causes uterine tocolytic effect without causing any cardiac toxicity or neuromuscular blockage; however, further human studies are desirable.

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The natural pyrethroids are not so much effective against agricultural pests, thus artificial synthesis of synthetic pyrethroids like fenvalerate was carried out by scientists. Fenvalerate is a insecticide has been proved to be highly toxic to non-target species like amphibians. In our study, Healthy frogs, Rana tigrina weighing 50 ± 3 gms were collected from the pond, acclimated to the laboratory conditions in large glass aquaria with water. The frogs were divided into groups of ten animals. They were exposed to different pesticide concentrations of fenvalerate, both commercial and technical grade, according to biomass ratio as suggested by Doundroff et al., (1951). The Total carbohydrate content in the present study Control Muscle tissue has shown higher amount of carbohydrates than compare with Brain and Intestine tissue. Under fenvalerate intoxication, the frog tissues showed consistent decrement in all the periods and it may be due to rapid utilization to withstand pesticide of stress. Total protein contents in control frog tissues the Muscle showed higher concentration of proteins than Intestine and Brain. The protein content in experimental frog tissues showed depleted levels when compared to controls. The significant decrease in protein and it may be due rapid utilization to withstand pesticide of stress. Total free amino acid (FAA) showed enhanced Intestine FAA levels, the levels in fenvalerate exposed frog tissues were increased significantly when compared to control. These elevated levels may be due to improved protease activity or enhanced transamination. From the above observations fenvalerate seems to be hazards to the aquatic life, causing severe biochemical changes and permanent architectural changes investigated in the present investigation.

Frog intestinal perfusion to evaluate drug permeability: application to p-gp and cyp3a4 substrates.

To evaluate the reliability of using in situ frog intestinal perfusion technique for permeability assessment of carrier transported drugs which are also substrates for CYP enzymes. Single Pass Intestinal Perfusion (SPIP) studies were performed in frogs of the species Rana tigrina using established method for rats with some modifications after inducing anesthesia. Effective permeability coefficient (Peff) of losartan and midazolam was calculated in the presence and absence of inhibitors using the parallel-tube model. Peff of losartan when perfused alone was found to be 0.427 ± 0.27 × 10-4cm/s and when it was co-perfused with inhibitors, significant change in Peff was observed. Peff of midazolam when perfused alone was found to be 2.03 ± 0.07 × 10-4cm/s and when it was co-perfused with inhibitors, no significant change in Peff was observed. Comparison of Peff calculated in frog with that of other available models and also humans suggested that the Peff-values are comparable and reflected well with human intestinal permeability. It is possible to determine the Peff-value for compounds which are dual substrates of P-glycoprotein and CYP3A4 using in situ frog intestinal perfusion technique. The calculated Peff-values correlated well with reported Peff-values of probe drugs. comparison of the Peff-value of losartan obtained with that of reported human’s Peff and Caco 2 cell data, and comparison of the Peff-value of midazolam with that of reported rat’s Peff, we could conclude that SPIP from model can be reliably used in preclinical studies for permeability estimation. This model may represent a valuable alternative to the low speed and high cost of conventional animal models (typically rodents) for the assessment of intestinal permeability.

Assessment of heavy metals and metalloids in tissues of two frog species: Rana tigrina and Euphlyctis cyanophlyctis from industrial city Sialkot, Pakistan.

In the present study, we investigated the concentrations of Ni, Fe, Pb, Cu, Co, Zn, Cd, Mn, and Cr in selected body tissues (liver, stomach, kidney, heart, lungs, and skeletal muscles) of two frog species: Rana tigrina and Euphlyctis cyanophlyctis captured from industrial wastewater of Sialkot city known worldwide for its tanning industry. The both frog species had darker appearance, distinctively different wet body weight, and snout-vent length. The results revealed that the heavy metal concentrations were high in the samples collected from industrial sites as compared to non-industrial sites. The different tissues of R. tigrina and E. cyanophlyctis exhibited little significant differences from two sites. The concentrations of heavy metals were more in tissues of R. tigrina as compared to E. cyanophlyctis. Mean concentration of Cd, Fe, Ni, Mn, Cu, and Cr was comparatively greater in R. tigrina, whereas Pb and Co were higher in E. cyanophlyctis. The concentration of Cu and Cd in the liver and kidney were relatively more in both species as compared to other organs. Further, the results indicated that frogs collected from industrial sites showed decreased body length and weight, and greater metal accumulation. The results will help the authorities for the conservation of these frog species which are under the influence of heavy metal contamination.

Comparative histologic anatomy of vertebrate liver

Introduction: The aim of the study was to compare the histological features of the liver of vertebrates and demonstrate the possible histological evolutionary trend within the sub-phylum vertebrata . Materials and Methods: Liver tissues were dissected from the five vertebrate classes; the Fish (Cat-fish - Clarias gariepinus ), Amphibian (Rana tigrina), Reptilian (Lizard - Agama aculeata ), Avian (Domestic bird- Gallus gallus domesticus ), and Mammalian (Wistar rat- Rattus norvegicus ) were used for the study. The tissue samples were fixed in 10% formal-saline and processed using standard histological techniques. The liver tissues were studied with a light microscope and photomicrographs were obtained. Results: The results showed structural similarities of the liver across the sub-phylum, with noticeable differences in the hepato-sinusoidal arrangements, composition and association of the bile duct and presence of portal triad. The study observed a gradual transition of hepatocytes distributions in singles and clusters to cord-like as observed in Gallus gallus domesticus and Rattus norvegicus . The study also revealed incomplete portal triad in Clarias gariepinus and Rana tigrina . The observed variations were attributed to adaption, feeding mechanism, mode of nutrition and selective specialization, which are important ecological and biological changes involved in evolution. Conclusion: This study has established that evolutionary trends occurred within the subphylum vertebrata.

Evaluation of Frog as an Animal Model to Study the Fraction of Oral Dose Absorbed in Humans

To evaluate single pass intestinal perfusion method in frog for the prediction of absorption in man and also to evaluate the predictability of this model to classify drugs correctly under biopharmaceutical classification system (BCS). Single Pass Intestinal Perfusion (SPIP) studies were performed in Frogs of the species Rana tigrina using established method for rats with some modifications. Permeability was determined for each selected drug in 6 frogs and the results were presented as mean ± SD. Effective permeability coefficient (Peff) of 12 drugs was calculated. Permeability assessment of some carrier transported drugs which are also substrates for CYP enzymes was also done. Correlation of frog Peff with fraction of dose absorbed (FA) in man was done. Rank order comparison of permeabilities obtained in this study with two other absorption models (human jejunal perfusion, rat intestinal perfusion) was made. The calculated Peff values correlated well with reported Peff values of probe drugs. Good correlation was found between Peff of humans and frogs (r2=0.942). The permeability classification based on frog intestinal permeability of drugs is in high agreement with previously introduced classification, and all the compounds are placed in correct categories as they belong to. The results of this study provided a basis that, as a permeability model in the early drug discovery stage, the single pass intestinal perfusion frog model can be used for the biopharmaceutics classification system. This model may represent a valuable alternative to the low speed and high cost of conventional animal models (typically rodents) for the assessment of intestinal permeability.

Genome-wide analyses of proliferation-important genes of Iridovirus-tiger frog virus by RNAi

Tiger frog virus (TFV), a species of genus Ranavirus in the family Iridoviridae, is a nuclear cytoplasmic large DNA virus that infects aquatic vertebrates such as tiger frog (Rana tigrina rugulosa) and Chinese soft-shelled turtle (Trionyx sinensis). Based on the available genome sequences of TFV, the well-developed RNA interference (RNAi) technique, and the reliable cell line for infection model, we decided to analyze the functional importance of all predicted genes. Firstly, a relative quantitative cytopathogenic effect (Q-CPE) assay was established to monitor the viral proliferation in fish cells. Then, genome-wide RNAi screens of 95 small interference (si) RNAs against TFV were performed to characterize the functional importance of nearly all (95%) predicted TFV genes by Q-CPE scaling system. We identified 32 (33.7%) genes as essential, 50 (52.6%) genes as semi-essential and 13 (13.7%) genes as nonessential for TFV proliferation. Quantitative RT-PCR and titer assays of selected genes were performed to verify the screen results. Furthermore, the screened essential genes were analyzed for their genome distribution and conservative comparison within Ranavirus. Such a systematic screen for viral functional genes by cell phenotypes should provide further insights into understanding of the information in antiviral targets, and in viral replication and pathogenesis of iridovirus.