ABSTRACT
Adiponectin receptor 1 (AdipoR1) and Adiponectin receptor 2 (AdipoR2) can bind to adiponectin (AdipoQ) secreted by adipose tissue to participate in various physiological functions of the body. In order to explore the role of AdipoR1 and AdipoR2 in amphibians infected by Aeromonas hydrophila (Ah), the genes adipor1 and adipor2 of Rana dybowskii were cloned by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed by bioinformatics. The tissue expression difference of adipor1 and adipor2 was analyzed by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and an inflammatory model of R. dybowskii infected by Ah was constructed. The histopathological changes were observed by hematoxylin-eosin staining (HE staining); the expression profiles of adipor1 and adipor2 after infection were dynamically detected by qRT-PCR and Western blotting. The results show that AdipoR1 and AdipoR2 are cell membrane proteins with seven transmembrane domains. Phylogenetic tree also shows that AdipoR1 and AdipoR2 cluster with the amphibians in the same branch. qRT-PCR and Western blotting results show that adipor1 and adipor2 were up-regulated at different levels of transcription and translation upon Ah infection, but the response time and level were different. It is speculated that AdipoR1 and AdipoR2 participate in the process of bacterial immune response, providing a basis for further exploring the biological functions of AdipoR1 and AdipoR2 in amphibians.
Subject(s)
Animals , Receptors, Adiponectin/metabolism , Phylogeny , Adiponectin/metabolism , Cloning, Molecular , Ranidae/geneticsABSTRACT
Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.
Subject(s)
Heat-Shock Proteins/genetics , Phylogeny , Amino Acid Sequence , HSP70 Heat-Shock Proteins/metabolism , Stress, PhysiologicalABSTRACT
RESUMEN La composición de la dieta constituye información básica sobre la historia natural de la especie. A pesar de la cantidad de datos adquiridos en los últimos años, aún queda mucho por conocer, especialmente para especies geográficamente extendidas. Aquí compilamos los elementos dietéticos disponibles de Leptodactylus vastus y reportamos el primer evento de depredación de Rupirana cardosoi por un juvenil de L. vastus. El hecho de que estas especies sean sintópicas en la región probablemente resultó en este nuevo evento de depredación. A diferencia de las observaciones previas de depredación de L. vastus, los especímenes que observamos no presentan una diferencia notable en el tamaño corporal, pero L. vastus fue capaz de casi tragar R. cardosoi, en coherencia con los hallazgos de que el tamaño de la boca está relacionado con la selección de presas en los anuros. Además, nuestra revisión de la literatura mostró que L. vastus es un depredador generalista y oportunista, que se alimenta de pequeños vertebrados (Amphibia, Squamata y Mammalia).
ABSTRACT Diet composition constitutes basic information on the natural history of the species. Despite the amount of data acquired in the last years, much remains to be known specially for geographically widespread species. Here we compiled the available dietary items of Leptodactylus vastus and report the first predation event upon Rupirana cardosoi by a juvenile L. vastus. The fact these species are syntopic in the region probably resulted in this novel predation event. Different from previous L. vastus predation observations, the specimens we observed do not present a striking difference in body size, but L. vastus was able to almost swallow L. cardosoi, coherent with findings that mouth size is related to prey selection in anurans. Also, our literature review showed that L. vastus is a generalist and opportunistic predator, that prey upon small vertebrates (Amphibia, Squamata, and Mammalia).
ABSTRACT
Leptin receptor overlapping transcript (LepROT) plays multiple roles in the regulation of immune systems. However, very little information is available about the anti-infectious mechanisms of amphibians LepROT. In this study, the cDNA sequence of the Rana dybowskii LepROT gene was determined by using RT-PCR and bioinformatics analysis. Then, the Aeromonas hydrophila (Ah) and lipopolysaccharides (LPS) infected models of R. dybowskii was constructed to obtain histopathological characteristics. Constitutive expression of LepROT mRNA and NF-κB signaling pathway were detected by real-time quantitative PCR. The full-length cDNA of LepROT gene was 396 bp and encoded 131 amino acids. Amino acid sequence analysis revealed LepROT shares 93.74% and 86.39% identity with homologues from other amphibians and mammals respectively, and the LepROT gene was quite conserved among different species. After infection, the relative expression levels of LepROT, NF-κB, IKKα and IKKβ mRNA were all significantly upregulated (P < 0.01), but showed a diverse temporal pattern of up-regulation in different tissues. Therefore, it was proposed that the LepROT gene of R. dybowskii might activate the NF-κB signaling pathway to exert anti-infectious effects, thus providing evidence for further extending the biological function of LepROT.
Subject(s)
Animals , Cloning, Molecular , DNA, Complementary , Gene Expression Profiling , Gene Expression Regulation , Mammals/metabolism , NF-kappa B/genetics , Phylogeny , RNA, Messenger/genetics , Ranidae/geneticsABSTRACT
Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I （<italic>CO</italic>Ⅰ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>， <italic>R. chensinensis</italic>， <italic>R. amurensis</italic>， <italic>R. culaiensi</italic>s， and <italic>R. huanrenesis</italic>， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>，<italic> Odorrana</italic>， <italic>Nidirana</italic>， <italic>Hylarana</italic>， and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>， <italic>R. zhengi</italic>， <italic>R. coreana</italic>， and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.
ABSTRACT
The evolution, structure and antigenic epitopes prediction of Rana dybowskii antimicrobial peptide dybowskin-1ST were carried out using bioinformatics software available online. Its antibacterial mechanism and structural properties were analyzed, and its activity was verified by applying wound healing assay in mice and bacteriostatic assay in vitro. This provides the theoretical basis for the improvement of parental peptide and the development of novel derivative peptides. The software MEGA_X were used to conduct homology alignment and to construct a phylogenetic tree. The online software ProtParam, ProtScale, PeptideCutter, signal, TMHMM Server were respectively used to predict the physicochemical parameters, hydrophilia/hydrophobicity, shear sites, signal peptides, and transmembrane domains of dybowskin-1ST. The online software SOPMA, Jpred4, DNAstar Protean were used to predict the secondary structure of dybowskin-1ST, and SWISS-MODEL, I-TASSER were used to predict the tertiary structure. ABCpred and SYFPEITHI were respectively used to predict its B-and T-cell epitopes. The effect of dybowskin-1ST on the wound healing was observed on experimental mice. Kirby-Bauer method and dilution method were used to determine the bacteriostatic activity of dybowskin-1ST. The dybowskin-1ST consists of 59 amino acid residues, of which leucine accounts for 16.9%, with a molecular formula of C₃₁₈H₅₁₀N₈₀O₉₃S₂. Its theoretical isoelectric point is 5.10 and the charge is -2. The dybowskin-1ST and dybowskin-1CDYa are closely related phylogenetically. The secondary structure of dybowskin-1ST predicted by the three methods were similar, which consisted of α-helix (44.07%), extended strand (16.95%), β-turns (3.39%), and random coil (35.39%). The prediction of tertiary structure showed that dybowskin-1ST was mainly composed of α-helix, and it was regarded as a hydrophilic protein with signal peptide sequence. Subcellular localization analysis showed that the probability of secreting the mitochondrial targeted peptides was 0.944. Dybowskin-1ST is an extracellular protein with no transmembrane structure region, but contains seven phosphorylation sites, three T-cell epitopes and eight B-cell epitopes. The dybowskin-1ST promoted wound healing and effectively inhibited the growth of Escherichia coli and Staphylococcus aureus. However, it had limited antibacterial activity against fungi and drug-resistant bacteria. Although the structure of dybowskin-1ST is rich in α-helix, the verification experiments showed that its antibacterial ability needs to be enhanced. The reason may be that it is a negatively charged and hydrophilic protein, and amino acid modification with the aim of increasing the number of positive charges and changing the hydrophobicity may be used to obtain derived peptides with enhanced activity.
Subject(s)
Animals , Mice , Amino Acid Sequence , Phylogeny , Pore Forming Cytotoxic Proteins , Protein Structure, Secondary , RanidaeABSTRACT
The aim of this study was to investigate the expression of MHCⅠ gene in different tissues of Rana dybowskii under the stress of Aeromonas hydrophila (Ah), and to provide evidence for revealing the anti-infective immune response mechanism of amphibians. The experimental animal model of Aeromonas hydrophila infection was first constructed, and the pathological changes were observed by HE staining. The MHCⅠ gene α1+α2 peptide binding region of Rana dybowskii was cloned by RT-PCR and analyzed by bioinformatics. Real-time PCR was used to detect the transcription level of MHCⅠ in different tissues under Ah stress. After Ah infection, the skin, liver and muscle tissues showed signs of cell structure disappearance and texture disorder. The MHCⅠ gene α1+α2 peptide binding region fragment was 494 bp, encoding 164 amino acids, and homology with amphibians. Above 77%, the homology with mammals was as low as 14.96%, indicating that the α1+α2 region of MHC gene was less conserved among different species. The results of real-time PCR show that the liver, spleen and kidney of the experimental group were under Ah stress. The transcript levels of MHCⅠ gene in skin and muscle tissues were higher than those in the control group at 72 h, but the time to peak of each tissue was different (P<0.01), indicating that the response time of MHCⅠ gene in different tissues was different under Ah stress. This study provides a reference for further exploring the immune function of MHC molecules in anti-infection.
Subject(s)
Animals , Aeromonas hydrophila , Gene Expression Profiling , Gene Expression Regulation , Allergy and Immunology , Gram-Negative Bacterial Infections , Allergy and Immunology , Liver , Metabolism , Ranidae , Genetics , Allergy and Immunology , Microbiology , Skin , MetabolismABSTRACT
Se revisa el estado taxonómico de Gastrotheca peruana usando métodos filogenéticos en base a secuencias de 16S rRNA. Los árboles de máxima verosimilitud y Bayesiano mostraron que las variantes génicas de G. peruana forman dos clados que no son hermanos. Uno de estos clados se distribuye en el norte de Perú, incluyendo un individuo procedente de la localidad típica de G. peruana dissimilis. El segundo clado está restringido al centro de Perú y contiene individuos de las localidades tipo de dos formas nominales, G. p. peruana y G. p. junensis, y es hermano de G. aratia. De esta forma, reconocemos dos especies dentro de lo que actualmente se conoce como G. peruana. Restringimos Gastrotheca peruana a las poblaciones del centro de Perú (departamentos de Ancash, Lima, Pasco y Junín) y asignamos Gastrotheca dissimilis a las poblaciones de los departamentos de La Libertad y Cajamarca.
The taxonomic status of Gastrotheca peruana is evaluated using phylogenetic methods and 16S gene sequences. The maximum likelihood and Bayesian trees showed that the genetic variants of G. peruana form two clades that are not sister to each other. One of these clades is present in northern Perú and includes a specimen collected at the type locality of G. peruana dissimilis. The other clade is restricted to central Peru and contains individuals from type localities of two nominal forms, G. p. peruana and G. p. junensis, and is sister to G. aratia. Our results suggest that G. peruana sensu lato currently encompass two species. We restrict Gastrotheca peruana to populations from central Perú (departments of Ancash, Lima, Pasco and Junín) and assign populations from departments of La Libertad and Cajamarca to Gastrotheca dissimilis.
ABSTRACT
El estudio se realizó en 13 zonas del litoral peruano del lago Titicaca, durante la época seca, desde junio hasta agosto de 2017. En cada zona se determinó la abundancia relativa y uso de microhábitat de Telmatobius culeus. En las zonas de evaluación se utilizó buceo con esnórquel para realizar en cada una tres transectos de 100 m x 2 m, con un total de 38 transectos evaluados. Se registraron 45 individuos en solo seis zonas de evaluación. El mayor número de individuos se encontraron en la zona de Perka Norte, con una abundancia relativa de 10.33 individuos/hora/02 personas a una profundidad de 1.5 m; en siete zonas no se registró ningún individuo. Los individuos fueron encontrados en los siguientes micro hábitats: 17 fueron encontrados sobre la vegetación acuática, 11 en arena combinado con vegetación acuática, 9 en roca combinado con vegetación acuática, 5 en fango combinado con vegetación acuática, 2 en roca y 1 en arena. a la vez se realizó análisis de diferentes parámetros de calidad de agua en cada zona de estudio, como: temperatura, pH, alcalinidad, dureza y oxígeno.
The Telmatobius culeus relative abundance and its microhabitat use was determined. In 13 areas of Peruvian littoral of Lake Titicaca was carried out, during the dry season, since June to August 2017. In each the evaluation areas, snorkeling was used to perform three 100 x 2 m transects. A total of 38 transects were evaluated. 45 individuals were registered in only six evaluation zones. The largest number of individuals were found in the North Perka area, with a relative abundance of 10.33 individuals/hour/02 people, at a depth of 1.5 m; in seven zones no individual was registered. Individuals were found in the following micro habitats: 17 were found on aquatic vegetation, 11 in sand combined with aquatic vegetation, 9 in rock combined with aquatic vegetation, 5 in mud combined with aquatic vegetation, 2 in rock and 1 in sand. At the same time, analyzes of different water quality parameters were performed in each study area, such as: temperature, pH, alkalinity, hardness and oxygen.
ABSTRACT
ABSTRACT Atelopus varius was believed to be extinct in Costa Rica until the rediscovery of a population within the Las Tablas Protected Zone (LTPZ) in 2008. This rediscovery represented a research and conservation opportunity, including contributing new information about the species' natural history. There are few reported species that prey on A. varius. This report presents a new predation record by a species of water bug (Abedus spp.) on an adult harlequin frog (A. varius). This is only the second confirmed predator for the species. These water bugs have been reported to prey on anurans during reproductive seasons, but in this case, the event occurred on A. varius out of their reproductive season. The effects of water bug predation on the only known reproductive population of this Critically Endangered species needs to be assessed to consider appropriate conservation measures to prevent further decline of the species.
RESUMEN Atelopus varius se consideró extinto en Costa Rica, hasta el redescubrimiento de una población en la Zona Protectora Las Tablas (ZPLT). Este hallazgo representa una oportunidad de investigación y conservación, incluyendo contribuir nueva información acerca de la historia natural de la especie. Hay pocas especies reportadas que depredan A. varius. Este reporte presenta un registro de depredación del chinche de agua (Abedus spp.), sobre un adulto de Sapo Arlequín (A. varius), como el segundo depredador confirmado de la especie. Usualmente, los chinches de agua depredan anuros durante su época reproductiva, pero este evento ocurrió fuera de la época reproductiva de A. varius. Los efectos de la depredación de chinches acuáticos sobre la única población reproductiva conocida de esta especie Críticamente Amenazada necesitan ser evaluados para considerar acciones de conservación.
ABSTRACT
The ovum oil of forest frog has various health beneficial functions. In the current research, we evaluated the hypolipidemic effects of the low-cholesterol ovum oil from the forest frog and its combination with stigmasterol in rats.
ABSTRACT
Objective:To establish an effective classification and identification method for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) maps of Rana dybowskii,its analogues and counterfeits based on cluster analysis and multivariate statistical analysis. Method:SDS-PAGE maps of 18 batches of R. dybowskii,its analogues and 2 counterfeits were obtained by SDS-PAGE method. SDS-PAGE maps were transformed into data matrix. NTSYSpc 2.10e statistical analysis software was used for cluster analysis,and SMICA-P 14.1 software was used for multivariate statistical analysis. Unsupervised Principal Component Analysis (PCA),Supervised Partial Least Squares Discriminant Analysis (PLS-DA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) were performed for multivariate analysis and evaluation. Result:SDS-PAGE maps technology combined with cluster analysis and multivariate statistical analysis could accurately classify and identify R. dybowskii,its analogues and counterfeits. Cluster analysis could cluster four kinds of medicinal materials into four branches except No.1 medicinal materials. PCA results were superior to cluster analysis. Supervised PLS-DA and OPLS-DA results in multivariate statistical analysis were superior to unsupervised PCA. The classification and identification efficiencies of OPLS-DA were better than those of unsupervised PCA. OPLS-DA aggregated R. dybowskii,its analogues and 2 counterfeits into four groups. Six different protein components were obtained by comprehensive analysis of variable importance in projection (VIP) value, and OPLS-DA Bi load diagram,with relative molecular weights were 51.363,35.838,14.565,17.563,15.358 and 21.696 kDa,respectively. Conclusion:SDS-PAGE maps combined with cluster analysis and multivariate statistical analysis can be used as an effective method to classify and identify R. dybowskii,its analogues and counterfeits. This study provides a reference for the quality evaluation and screening of R. dybowskii.
ABSTRACT
Ranae Oviductus has a high economic and social value, but its adulterants are more numerous, which causes a great confusion to the market. Using DNA bar code technology based on COI sequence for PCR amplification and sequencing of the identified Rana dybowskii, R. chensinensis, R. huanrensis and R. amurensiss, the COI gene database of four species of Rana was established, and comparing the measured sequence with the sequence of GenBank, four kinds of Rana were identified. The MEGA (molecular evolutionary genetics analysis) 7 .0 software was used to calculate the genetic distance of K2P and construct the NJ (neighbor-joining) system cluster tree. The sequence of the four species of Rana measured were clustered into one group with the sequence of the four kinds of Rana downloaded from GenBank, but separated from the two outer groups downloaded from GenBank. The COI gene of the R. dybowskii was likely to have regional differences, however this technique failed to distinguish male and female Rana. The results showed that DNA bar code technology could accurately identify the base of original animal of R. oviductus. It indicates that DNA bar code COI provides a new method for the identification of R. oviductus.
ABSTRACT
Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles performance, jeopardizing their survival.
Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.
ABSTRACT
Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L–1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles’ cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles’ performance, jeopardizing their survival.
Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L–1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration – MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.
Subject(s)
Animals , Heart/drug effects , Insecticides/toxicity , Methyl Parathion/toxicity , Rana catesbeiana/physiology , Heart/physiopathology , Larva/drug effects , Larva/growth & development , Larva/physiology , Rana catesbeiana/growth & development , SwimmingABSTRACT
Background: Different species of frogs had been used by many researchers for evaluation of analgesic drugs e.g., Rana pipiens and African claw frog. In our study, we used Rana tigrina, which was never used for evaluation of analgesic activity of drugs. So by doing this project, we judged usefulness of R. tigrina to evaluate analgesic drugs. Methods: Animals used were R. tigrina of either sex weighing 100-150 g. Glass flask with porous platform was used for observation of frog. All groups were treated with 4% NaCl solution S.C. on abdomen). Characteristic parameter i.e., number of eye blinkings (this parameter was observed during the pilot study after 4% NaCl S.C. injection on lower third of frogs abdominal wall) were observed before and after drug administration. Each observation was for 5 mins. Centrally and periphrally acting drugs effect was tested on the number of blinks and buccal oscillations. Results: Centrally acting drug inhibit rise in number of eye blinkings and buccal oscillation significantly. Piroxicam diclofenac sodium and ketoprofen decreased rise in number of blinks; but, it was not significant as compared with control. Conclusion: This animal may be used alternative to existing methods of evaluating analgesics in Indian setup.
ABSTRACT
Geobatrachus walkeri is a frog that belongs to a monotypic genus and is endemic to the Sierra Nevada de Santa Marta, Colombia. The species inhabits different microhabitats between 2 000 to 3 500m altitude, including the leaf litter of a pine plantation. To understand its reproductive ecology, we conducted eight frog samplings, covering the rainy and dry seasons, and two habitat types (secondary native forest and pine plantation) during 2010-2011. For this study, we also included data obtained from five previous similar samplings undertaken during 2008-2009. The pine leaf litter was the main microhabitat where frogs were found; we heard choruses of six-ten calling males during all sampled months, and observed the frogs having diurnal and nocturnal activity. Regardless of the year of study, the population consisted of neonates recruited several times of the year, a large number of juveniles with a wide range of body sizes, and fewer adults with a narrower range of body size. The histological analyses of the gonads showed that the size at maturity was near 18mm SVL for males and females, and those adult males and females were reproductive active during all sampling months, suggesting a continuous reproductive activity pattern. However, during the dry season, the seminiferous tubules showed a drastically diminished spermatic epithelium although containing abundant luminal spermatozoa, which suggest a reduction in the sperm production at the end of this season. Similarly, frogs of all age categories were significantly more abundant during the early dry season, whereas were significantly less abundant with the advancement of the dryness suggesting that the intensity of the dry season could temporally stop the activity and reproduction of this population.
Geobatrachus walkeri es una rana que pertenece a un género monotípico y es endémica de la Sierra Nevada de Santa Marta, Colombia. La especie habita en diferentes microhábitats entre los 2 000 y 3 500m de altitud, incluyendo la hojarasca de una plantación de pinos. Para entender su ecología reproductiva se realizaron ocho muestreos de individuos, que abarcaron las estaciones de lluvias y de sequía y dos tipos de hábitats (bosque secundario y plantación de pino) durante 2010-2011. También se incluyeron los datos de cinco muestreos similares obtenidos previamente durante el período 2008-2009. El principal microhábitat donde las ranas se encontraron fue la hoja-rasca de pino, allí escuchamos coros de seis-diez machos durante todos los meses muestreados, y observamos que las ranas tienen actividad diurna y nocturna. Sin importar el año de estudio, la población se compone de neonatos reclutados en varios meses del año, un gran número de juveniles con un amplio rango de tamaños corporales, y un menor número de adultos con un rango más reducido de tamaño corporal. Los análisis histológicos de las gónadas mostraron que el tamaño a la madurez se adquiere cerca de los 18mm LRC para machos y hembras, y que los machos y hembras adultos fueron reproductivos durante todo el muestreo, lo que sugiere un patrón continuo de actividad reproductiva. Sin embargo, durante la estación seca, los túbulos seminíferos presentan un epitelio espermático drásticamente disminuido, aunque contienen abundantes espermatozoides luminales, lo que sugiere una reducción en la producción de espermatozoides en el final de la estación seca. Del mismo modo, las ranas de todas las categorías de edad fueron significativamente más abundantes durante el inicio de la estación seca, mientras que fueron significativamente menos abundantes con el avance de la sequedad, lo que sugiere que una mayor intensidad de la temporada de sequía podría detener temporalmente la actividad y la reproducción en esta población.
Subject(s)
Animals , Female , Male , Anura/physiology , Ecosystem , Sexual Maturation/physiology , Anura/classification , Colombia , Gonads/anatomy & histology , Gonads/growth & development , Reproduction/physiology , SeasonsABSTRACT
Background: Rana Plaza collapse is the worst and deadliest man-made industrial disaster in the history of garment sector in the world. Around 1200 people died and thousands more were injured. Most of the victims of the disaster were treated in Enam Medical College Hospital (EMCH). We conducted this study to give an overview on the disaster victims and services provided by EMCH. Objective: The study was done to observe the length of time between accident and admission in EMCH, length of time between admission and discharge, to observe the numbers, types and sites of injuries to the victims, medical measures given to the victims and finally to observe the status of the victims at the time of discharge. Materials and Methods: This descriptive type of observational study was carried out among the victims of Rana Plaza tragedy during the period of May to October 2013. All admitted patients in EMCH were included in the study. Purposive nonprobability sampling technique was applied in this research work. Data were collected from the hospital record. After collection, data were manually compiled, edited and analyzed. Results: Among 621 victims treated in different wards, 276 (44.45%) were admitted to the hospital on the day of accident. Among the admitted patients, 255 (41.06%) stayed in the hospital for 1--3 days,133 (21.42%) for 4--7 days and 88 (14.17%) for more than 10 days. Fracture and dislocation were present in 32.70% patients, lacerated injury in 18.20%, abrasion in 15.78%, bruise in 13.53%, incised wound in 15.45% and punctured wound in 4.34% patients. Single injury was present in 56.68% and rest had multiple injuries. Lower limbs were the most (33.01%) affected part of the body followed by head and neck (22.06%), upper limbs (18.52%), thorax (17.55%) and abdomen (8.86%). Two hundred seventy two patients (43.80%) were improved after treatment and 56 (9.02%) were fully cured; 23.83% of the victims were referred to higher medical centers for special measures. Conclusion: The collapse of Rana Plaza has fueled a greater call for reform and safety in garment industry. At the same time, all health care centers should adopt a broad-based approach to disaster-preparedness by providing a framework to ensure a well-coordinated response to mass casualty event.
ABSTRACT
A new trichostome ciliate, Balantidium honghuensis n. sp., was isolated from the recta of Rana nigromaculata and R. limnocharis during parasite surveys in Honghu Lake, Hubei Province, central China in summer of 2010 and 2011. Its detailed morphometric characters based on LM and SEM studies were described herein. The organism is oval in shape and thickly ciliated. The vestibulum is "V" shaped and occupies about 1/3 to 2/5 of the body length. The vestibular and nearby regions possess strong peripheral fibers which form a marked axial fiber about the cytopharynx. More than 10 contractile vacuoles are distributed along the periphery of the latter body. Comparisions were made between this new species and B. sinensis Nie, 1935. They were discriminated from each other in terms of general body forms, body size, and vestibulum shapes. Besides, special attention was paid to its high-speed daughter swarmers which we believed to be the infective stage of B. honghuensis. Possible infection routes of anura amphibian balantidia were discussed.
Subject(s)
Animals , Balantidiasis/parasitology , Balantidium/classification , China , Ranidae/parasitology , Rectum/parasitologyABSTRACT
The most important use of neuromuscular blockers is as adjuvant to general anaesthesia where adequate muscle relaxation can be achieved at lighter plane. They also reduce reflex muscle contraction in the region undergoing surgery and assist maintenance of controlled ventilation during anaesthesia. They are particularly helpful in abdominal and thoracic surgery, intubations and endoscopies, orthopedic manipulation, etc. Thus, the risk of respiratory and cardiovascular depression is minimized, and post anesthetic recovery is shortened. The potency ratio of two commonly used neuromuscular agents depolarizing succinylcholine and non depolarizing pancuronium. Double pith a frog and fasten it to a frog board with ventral side up. The sternum was cut through just above the xiphisternum at its base and a pair of muscle attached to it were dissected out and transferred to a dish containing frog Ringer solution at room temperature. All the drug containing solutions were freshly prepared before the experiments Succinyl choline, Pancuronium (1,10,100mg/ml and 1mg/dl) respectively Acetyl choline (10,100mg/ml and 1mg/dl). Acetylcholine solution in various strength were prepared starting from 0.1% to 0.0001%.NMJ blocker Pancuronium was added to the biophase in addition to selected dose (128mg or 256mg) and the contraction of muscle till the 70-80% of inhibition is produced and the difference from sub maximal contractions. The median ED50 was interpolated from the figure taking 50%of inhibition from Height of contraction in mm. The ‘t’ test was performed to compare the ED50 value were interpolated from the regression line to find out the ED50 of the drug. The median doses (ED50) of both of them were calculated graphically and compared. The mean ED50 value of succinylcholine was found to be 1.59 ± 0.08μg (95% confidential limit was from 1.53 to 1.66μg). The ED50 of pancuronium was found to be 0.52 ± 0.10μg with 95% confidence limit being from 0.44 to 0.60μg. The ED50 value of the two drugs was very significantly different (P < 0.001). The potency ratio of pancuronium to succinylcholine was 0.32.