Pelophylax Bedriagae Research Articles

Adeno-associated viral tools to trace neural development and connectivity across amphibians.

Amphibians, by virtue of their phylogenetic position, provide invaluable insights on nervous system evolution, development, and remodeling. The genetic toolkit for amphibians, however, remains limited. Recombinant adeno-associated viral vectors (AAVs) are a powerful alternative to transgenesis for labeling and manipulating neurons. Although successful in mammals, AAVs have never been shown to transduce amphibian cells efficiently. We screened AAVs in three amphibian species-the frogs Xenopus laevis and Pelophylax bedriagae and the salamander Pleurodeles waltl-and identified at least two AAV serotypes per species that transduce neurons. In developing amphibians, AAVs labeled groups of neurons generated at the same time during development. In the mature brain, AAVrg retrogradely traced long-range projections. Our study introduces AAVs as a tool for amphibian research, establishes a generalizable workflow for AAV screening in new species, and expands opportunities for cross-species comparisons of nervous system development, function, and evolution.

Hazard assessment of microplastics and heavy metals contamination in Levant frogs (Pelophylax bedriagae): A bioindicator in Western Iran

The increasing pollution of aquatic ecosystems caused by microplastics (MPs) and heavy metal ions worldwide threatens the life of aquatic organisms, including amphibians. In this study, we investigated the presence and accumulation of MPs and heavy metal ions in the upper gastrointestinal tract (GIT) of the Levant frog (Pelophylax bedriagae) as a bioindicator in contaminated sites of Western Iran. A total of 192 adult frogs from 16 locations in the west and northwest of Iran were collected. We measured the accumulation of MPs and heavy metal ions in the collected frogs and determined the characteristics of MPs in the samples. Our findings revealed widespread MPs and heavy metal ions contamination in the frog GIT across all sampled stations. We found 742 MP particles in the digestive system of frogs, with the highest (7.09 MP/individual) and lowest (2.37 MP/individual) number observed in stations 10 and 9, respectively. Fibers were the most common MPs and polyester (PES) (42.5%) and polyethylene terephthalate (PET) (17.2%) were found to be the most common polymers in the GIT of frogs. The average accumulation of heavy metal ions showed significant differences (P < 0.05) among 16 sampling stations, and zinc (II) and mercury (II) showed the highest and lowest concentrations in frogs. We also found moderate to strong positive correlations between the detected MPs and heavy metal ions in the frog samples across all sampling stations. Our findings confirm the contamination of frogs by MPs and heavy metal ions and the potential capacity of MPs to increase the toxicity of heavy metals in P. bedriagae as a bioindicator in contaminated sites.

Investigation of heavy metals in tissues and habitats of three edible frogs from Türkiye.

The soil, water, and organisms have been contaminated by heavy metals due to human activities and industrialization, which has produced a major environmental problem that has a deleterious effect on human health and food quality. Frogs, one of the good bioindicators for environmental pollution, are also among the alternative essential protein sources for humans. In Türkiye, three of these frogs are edible: Pelophylax ridibundus, Pelophylax bedriagae, and Pelophylax caralitanus, also known as Anatolian water frogs. Hence, to assess the possible health risks that might result from consuming frog legs in addition aquatic habitat of Anatolian water frogs, the water, sediment, and frog tissue samples (muscle and liver) were obtained from 11 different provinces covering all regions of Türkiye and analyzed to determine Cd, Cu, Cr, Zn, Pb, and As concentrations. The results revealed considerable variations in heavy metal concentrations among frog tissues, influenced by the sampling sites and species (ANOVA: p < 0.05). The Estimated Daily Intake (EDI) values, calculated based on the average serving size, were also lower than the Provisional Tolerable Daily Intake (PTDI) levels for adult consumers. Furthermore, the study computed the Target Hazard Quotient (THQ) values for heavy metals, all of which were below the critical value of 1, indicating that consuming the hind leg muscles from these frog species would not pose an adverse health risk for humans.

Skeletochronology of the Levant Green Frog, Pelophylax bedriagae (Camerano, 1882), in Southern Iran

Skeletochronology is a reliable method for estimating the age and growth rate of many ectothermic vertebrates, including amphibians. The Levant green frog, Pelophylax bedriagae (Ranidae), is a relatively large frog distributed in the western and northern parts of Iran. The age structure of Pelophylax bedriagae was determined by examining 52 females and 39 males collected from Kohgilouyeh and Bouyerahmad Provinces (Yasouj and Basht). The age range of the two populations is three to 10 years. The maximum age for males and females is 10 and eight years, respectively. The highest age distribution for males is six years (17.6%) and four and six years (15.4%) for females. On average, males (6.2±1.4 years) are older than females (5.2±1.3 years). Male and female body size is correlated with age within both populations. However, despite being significantly different in average body size, the average age of the two populations (Yasouj vs. Basht) is not significantly different.

Skeletal description of Pelophylax caralitanus (Anura: Ranidae): Comparison with other ranids.

A description of the skeletal development of ranids is scarce despite being well known in the family Ranidae. Herein, the description of several species of two genera representing the family Ranidae from Turkey is studied wherein the larval and adult cranial skeletons of Pelophylax caralitanus are described and compared with that of the water frog (Pelophylax bedriagae) and mountain frog (Rana macrocnemis). The data are based on cleared and double-stained specimens of 20 Gosner developmental Stages (26-46). The first element to ossify in P. caralitanus is the parasphenoid (Stage 30), followed by the exoccipitals (Stage 34) and prootics (Stage 35). The metamorphic climax commences at Stage 42 during which several modifications to the chondrocranium will unfold. In addition, ranids demonstrate remarkable differences between the water (Pelophylax sp.) and mountain (Rana sp.) frog species: (a) the ventromedial bridge between the corpora of the suprarostral cartilage in the larval upper jaw, (b) the number of processes of the central corpus in the hyobranchial skeleton, and (c) the differences in the ossification time and sequence between Pelophylax sp. and Rana sp. A detailed description of the larval osteological features of P. caralitanus conforms to the current phylogenetic position and provides a model for comparison with other ranids.

Microplastic prevalence in anatolian water frogs (Pelophylax spp.)

Frogs are on the verge of extinction due to various biotic and abiotic stressors. Rivers, lakes, ponds, wetlands, and ditches that make up their habitats are exposed to different anthropogenic pollutants. Today, plastics stand out among these pollutants due to their widespread use; however, the information on microplastic (MP) accumulation in frogs is insufficient. In the present study, adult frog samples were collected from 19 different stations in Türkiye, including marsh frogs (Pelophylax ridibundus) from 18 stations and levantine frogs (Pelophylax bedriagae) from 1 station. MP was found in 147 (82.4%) of the 176 frogs that were analyzed. MP abundance in frogs varied between 0.20 and 18.93 MP individual−1. The characterization of MPs was determined predominantly as follows; PET (70.1%) in polymer type, fiber (92.2%) in shape, navy blue/blue (76.1%) in color, and >90% were smaller than 300 μm. No significant relationship with frog's weight (correlation coefficient = 0.01, P = 0.812) or length (correlation coefficient = 0.06, P = 0.473) and MP abundance was detected. The outcomes of this survey might be a baseline to assess the ecological risks posed by MPs and to guide future experimental research.

Hematology of the Levant Green Frog, Pelophylax bedriagae (Amphibia: Ranidae) in southern Iran

In this study, 68 specimens of the Levant Green Frog, Pelophylax bedriagae, were collected from agricultural fields around Yasouj and Noor-Abad cities, from August 2018 to May 2019 in southern Iran to investigate the blood cell morphology. Blood specimens were taken from the heart, or abdominal and facial veins. Smears were dried in room temperature, fixed by methanol and stained using Giemsa method. Blood cells (erythrocytes and leucocytes) were counted and measured using light microscopy under an optical micrometer. Based on the findings of this study, erythrocytes from Noor-Abad population has significantly greater values from those in Yasouj population in all of the six metric characters (P < 0.05). RBC’s in male frogs are also greater than those in females in all six studied characters (P < 0.05). All studied metric characters in lymphocytes are also different between tow localities and sexes (P < 0.05).

Characterization of testicular histology and spermatogenesis in the Levantine frog, Pelophylax bedriagae (Amphibia: Anura: Ranidae)

Amphibians occupy a position of great interest in terms of vertebrate evolution. Additionally, amphibians are known as a transitional group between amniotes and anamniotes. However, there are few studies on the gametogenesis of anamniotes vertebrates, especially anurans. Therefore, the purpose of this study was to analyze the histological feature of germ cells and their arrangement in the testis of Levantine frog, Pelophylax bedriagae (Camerano, 1882). Spermatogenic cells were organized in spermatocysts. Each spermatocyst contained cells at the same stage of the spermatogenic cycle. Identification of each cellular type in seminiferous tubule was carried out according to the size and morphology of cells and the degree of nuclear material compaction. Spermatogonia were large cells localized at the base of the seminiferous epithelium. Primary spermatocytes were examined in different phases of first meiotic division and distinguished from other cell types by their dark spherical nuclei or looser chromatin. Two types of spermatids, spherical and elongated cells, were observed. Seminiferous tubules were surrounded by peritubular myoid cells, and they contained no lumen. The lack of lumen in the seminiferous tubules and the cystic spermatogenesis probably provide synchronously production of a large number of sperms. The location of hyaluronic acid was also determined in interstitial tissue between seminiferous tubules to probably provide testicular integrity and viscoelasticity.

Prevalence and pathology of Lernaea cyprinacea (Crustacea: Lernaeidae) parasitizing tadpoles of Pelophylax bedriagae (Anura: Ranidae) in Antalya, Turkey

Prevalence and pathology of Lernaea cyprinacea (Crustacea: Lernaeidae) parasitizing tadpoles of Pelophylax bedriagae (Anura: Ranidae) in Antalya, Turkey. This is a report of the frst record of the occurrence of Lernaea cyprinacea on tadpoles of Pelophylax bedriagae, a frog species native from Karpuzçay Creek, Antalya, Turkey. A total of 20 tadpoles of P. bedriagae were examined and 14 (70%) specimens were infected. A maximum of two parasites per host was found, primarily around the cloaca. In general, slight to moderate hyperemia was noticed around the parasite. Typically, granuloma form in the tissues around the parasite. Histopathological examination revealed that hemorrhages and pathological lesions such as dermal erosions, and a chronic infammatory reaction in skin and muscle were present at the site of parasite attachment.

Life history of the Levant water frog, Pelophylax bedriagae (Amphibia: Anura: Ranidae) in western Iran

In the present study, we studied the development and morphology of different larval stages of Pelophylax bedriagae (Camerano) at two rearing temperatures (20 and 24 °C). Eggs were collected from a breeding site in western Iran. Diagnostic morphological characters are provided for Gosner (1960) larval stages 1–46. The larvae hatched about seven days after egg deposition in the laboratory. A principal diagnostic feature, the formation of the funnel-shaped oral disc, became discernible about ten days after hatching at Gosner stage 21 and degenerated at Gosner stage 42. Based on our results, the longest metamorphosis time was observed at 20 °C whilst the shortest metamorphosis time occurred at 24 °C. The largest body length of larval P. bedriagae measured about 54 mm in 70 days after egg deposition. Compared with the majority of other Palearctic anurans, it appears that embryonic and larval development is usually slow in P. bedriagae.

Histological and histochemical study on the mesonephric kidney of &lt;i&gt;Pelophylax&lt;/i&gt; &lt;i&gt;bedriagae&lt;/i&gt; (Anura: Ranidae)

The purpose of this study was to determine the histological structure of the kidney of Pelophylax bedriagae and the distribution of hyaluronic acid (HA) in the kidney tissue. The kidneys of P. bedriagae are long, brown, ribbon-like structures covered with connective tissue. Nephrons, the functional units of the kidney, consist of the renal corpuscle, proximal and distal tubules, collecting tubule, and collecting duct. The renal corpuscle is composed of the glomerulus and Bowman?s capsule. There is no structure similar to Henle?s loop, which is found in higher vertebrates. The visceral layer of Bowman?s capsule is composed of podocytes surrounding the glomerular capillaries. The parietal layer of Bowman?s capsule is lined with a simple squamous epithelium. The proximal tubule is formed by cubic epithelial cells with a brush border, and the distal tubule is covered with a simple cubic epithelium without a brush border. The collecting ducts consist of columnar or cubic cells, and they are larger than the proximal and distal tubules. Many melanomacrophage centers were observed in the kidney parenchyma. The localization of HA was determined to be in the interstitium surrounding the collecting ducts. HA probably plays a significant role in renal water handling and electrolyte balance due to its ability to retain water and bind cations.

Age structures and growth parameters of the Levantine frog, Pelophylax bedriagae, at different localities in Denizli, Turkey

Skeletochronology is a reliable tool for assessing several parameters in amphibian populations. We used skeletochronology to determine the age structure, growth rate, age at first reproduction, and longevity of Levantine frog Pelophylax bedriagae populations from different localities in Denizli, Turkey. All examined individuals (N = 161) exhibited Lines of Arrested Growth in the bone cross-sections. Age structure and age at first reproduction were similar among localities and sexes, while longevity and growth rates showed significant variation among localities. Obtained results were compared with literature data on age-related and grow parameters in Pelophylax bedriagae and cognate species.

New insights on the phylogenetic position and population genetic structure of the Critically Endangered Karpathos marsh frog Pelophylax cerigensis (Amphibia: Anura: Ranidae)

The genus Pelophylax , which currently comprises 26 species, is a well studied group due to its complex history and high diversification, although some phylogenies remain unresolved. Here we assess the phylogenetic position and the population genetic structure of the Critically Endangered Karpathos frog, Pelophylax cerigensis , endemic to Karpathos Island. A total of 42 cytb sequences were examined including specimens from Rhodes Island, and amplified fragment length polymorphisms (AFLPs) were generated to investigate the genetic structure and connectivity of the only two known populations on Karpathos. Molecular analyses reveal two major waterfrog lineages in the eastern Mediterranean: Clade A comprises Pelophylax bedriagae from the Middle East and the island of Cyprus, while clade B includes both P. bedriagae from the east Aegean Sea and P. cerigensis . Specimens from Karpathos and Rhodes constitute a single clade, contrasting previous studies, thus indicating the occurrence of Karpathos frog also to the neighboring Rhodes. The AFLP markers revealed low but statistically significant levels of genetic divergence between the two Karpathos’ populations and similar levels of genetic diversity. Our results suggest that the current taxonomy of the species should be re-evaluated. We also strongly recommend the need of conservation actions to maintain the levels of diversity in the declining population of the Karpathos frog.

Taxonomic survey of water frog populations of Pelophylax bedriagae (Anura: Ranidae) in western Iran: a morphometric and bioacoustic approach

We explored the population systems of Pelophylax bedriagae (Camerano, 1882) in northwestern, western and southwestern Iran. Statistical significance for a hypothesis of sexual dimorphism of the frogs was tested using t-test and multivariate analysis of variances (MANOVA). Due to lack of sexual differences between the male and female frogs, the data of both sexes were pooled for further analysis. Taxonomic relationships among local populations were investigated using discriminate analysis (by stepwise method) with iterative sample assignment. The multivariate morphometric analyses revealed that the range of P. bedriagae extends to western Iran. The next step was to analyze the advertisement call (five standard parameters: call duration, number of pulse groups per call, duration of pulse groups, interval between pulse groups, and intercall interval) of the water frogs inhabiting three sites in western Iran. Rigorous application of the bioacoustic method removes all doubt that these three sites are inhabited by the same species, which on the basis of priority of nomenclature should be called P. bedriagae.

Histopathological analysis of carbaryl-induced toxicity in the spleen of Levantine frog, Pelophylax bedriagae (Anura: Ranidae).

This study describes the histopathological effects of carbaryl in the spleen of Levantine frog, Pelophylax bedriage. Due to its primary role in immune system, it is important to research the toxic effects of pesticides, which play an important role in environmental pollution, on spleen. To that end, adult frogs were exposed to carbaryl for 96h. Experimental groups contained low dose, (0.05mg/g), medium dose (0.1mg/g) and high dose (0.2mg/g). After following exposure to carbaryl, the frogs were euthanised and dissected. In low-dose group, there were no important changes in spleen tissue. In medium-dose group, prominence in haemorrhage just below the capsule and an increase in the melanomacrophage number were determined. In high-dose group, in addition to increases in the melanomacrophages, separations in capsule, haemorrhage below capsule and within splenic tissue, sinusoidal enlargement, congestion in dilated sinusoid, hypertrophic plasma cells and fibrosis were determined as important histological lesions in exposed frogs. This study clearly showed that carbaryl caused important histopathological damages in splenic tissue of Pelophylax bedriagae. In view of these findings, it can be said that this insecticide has the capacity to disrupt spleen' functions.

First bioacoustic and morphological data for the presence of Pelophylax bedriagae in Bulgaria

Abstract Water frog mating calls from two localities were studied and analyzed. Recordings were made in the summer of 2010 at the Arkutino swamp near the town of Primorsko and at the Vurbitza River near the town of Momchilgrad. A total of 154 calls were analyzed and the results suggested the presence of both the Marsh frog (Pelophylax ridibundus) and the Levant frog (Pelophylax bedriagae) in both sites, with the former being more frequent in Vurbitza River, and the latter – in Arkutino. At Vurbitza, we also captured and measured 2 specimens, which morphological characteristics differed from P. ridibundus and matched those of P. bedriagae. These are the first localities for P. bedriagae in Bulgaria.

Natural history and conservation of the rediscovered Hula painted frog, Latonia nigriventer

Dramatic global amphibian declines have recently led to an increased concern for many species of this animal class. The enigmatic Hula painted frog (Latonia nigriventer), the first amphibian to be declared extinct but unexpectedly rediscovered in 2011, has remained one of the rarest and most poorly understood amphibians worldwide. Gathering basic biological information on this species, along with an understanding of its disease-related threats remains fundamental for developing risk assessments and conservation strategies. Our surveys in recent years confirmed that L. nigriventer is a localised species with elusive habits. The species appears to follow an opportunistic breeding phenology and has a tadpole morphology similar to its well-studied sister group Discoglossus. However, the adults’ extended annual presence in the aquatic habitat is a major difference from species of Discoglossus. We detected the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), in northern Israel and on Hula painted frogs but did not observe any signs of chytridiomycosis in this species. Our preliminary data on aspects of the innate immunity of L. nigriventer suggest that the skin mucosome of this species contains antimicrobial peptides and a bacterial community differing from other syntopic frogs (Pelophylax bedriagae). The combined knowledge of both natural history and innate immunity of L. nigriventer provides valuable insights to direct future research and conservation management of this critically endangered frog species.

A Note on the Karyotype of the Amphibian Pelophylax Bedriagae from Jordan

A Note on the Karyotype of the Amphibian Pelophylax Bedriagae from Jordan

Life Cycle of Tree Frogs (Hyla savygnyi) in Semi-Arid Habitats in Northern Israel

The life cycle of tree frogs (Hyla savygnyi) in localities of various habitats in northern Israel in the Upper Galilee and Golan Heights (annual rainfall range of 500-1,000 mm) and the Hula Valley, ranging from 212 to 740 m above sea level (ASL), was studied. Tree frogs were observed around winter rain pools, springs and streams. Fifty-one different breeding places were monitored. Only ponds and springs having stable non-flowing water are used by tree frogs for reproduction and are where larvae can metamorphose. The male call in the breeding places attracts the female, who then comes to the male, and breeding occurs underwater. Breeding time in Israel varies according to location and geographical and ecological conditions. It takes place in March-April in northern Israel, the Upper Galilee, the Golan Heights and the Hula Valley. In various breeding places, larvae grow between April and June. Apparently tree frog larvae adapt to breeding places where stable water is found, such as in ponds or springs, but not to breeding places with flowing water such as streams or rivers. Various Anuran and Urodela larvae were found in the same breeding sites where the larvae of H. savygnyi were found, including Pseudepidalea Viridis, Rana bedriagae, Pelobates syriacus, Salamandra infraimmaculata and Triturs vittatus. Following metamorphosis, H. savygnyi adapt to terrestrial life in semi-arid habitats, and is found on plants or in hiding places under rocks and in holes to prevent dehydration particularly during hot and dry weather. The ability to change color helped the frogs to hide in various substrates. The rate of water loss of terrestrial H. savygnyi during dehydration is around 50% of body weight. Plasma osmolality increased from 200 mOsm/kg to about 450 mOsm/kg, which helped the tree frog to survive in semi-arid habitats.

Helminth parasites of the levantine frog (Pelophylax bedriagae Camerano, 1882) from the western part of Turkey

Summary Fifty-four Pelophylax bedriagae (Levantine Frog) from Turkey (İzmir and Manisa Provinces) were examined for helminth parasites. The frogs were collected between 2012 and 2014 years. Eight species of helminth parasites were recorded: 3 species of Nematoda (Rhabdias bufonis, Cosmocerca ornata, Oswaldocruzia filiformis), 3 species of Digenea (Diplodiscus subclavatus, Haematoloechus bre-viansa, Gorgoderina vitelliloba), 1 species of Acanthocephala (Acanthocephalus ranae) and 1 species of Hirudinea (Hirudo medicinalis). Pelophylax bedriagae is a new host record for these parasite species.