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We describe the advertisement call of the Nanling horned frog, Boulenophrys nanlingensis for the first time, based on recordings obtained from four individuals. One of these individuals, which was identified using its dorsum pattern, was recorded twice after nine months. Distinct shifts in the temporal parameters and call rate were observed from calls of the re-captured individual, which was suspected to be related to temperature and social context. However, due to the limited sample size, further research is needed to confirm these findings. We highlight the potential of mark-recapture method using dorsum pattern for studying and monitoring the Nanling horned frog and other megophyinid frogs.

Based on debilitating recent budget cuts for science, Brazilian researchers had to find alternative ways to continue scientific production. Here we provide a perspective for the use of citizen-science data deposited in the iNaturalist platform as an alternative source of data to support biodiversity research. Observations contributed by volunteers can be analyzed at large spatial and temporal scales and can respond to questions in behavioral and population ecology. We analyzed this potential through the example of Brazilian amphibians, a group that is less studied worldwide than birds. In fact, to our knowledge, only two studies have been published that are based on citizen-science data for Brazilian amphibians. At the time of writing, the iNaturalist platform has over 14,800 research grade observations from Brazil, representing 698 species, a number increasing daily. Compared to other species-rich countries, volunteer-collected datasets from Brazil cover a relatively high taxonomic diversity (61%), providing a plethora of valuable data. Despite this potential, there are large spatial gaps in sampling in Brazil. Here we encourage established and budding herpetologists not only to use the platform to retrieve data, but also to contribute to iNaturalist actively, with new observations, as well as by identifying species in existing records.

The present study describes a rich amphibian and reptilian assemblage from the early Miocene locality Echzell, Germany. It consists of one allocaudate, five salamander, five frog, one gecko, chamaeleonids, anguine lizards, one lacertid, one skink and five snake taxa. The entire herpetofauna of Echzell is represented by genera and/or families very broadly known from the early Miocene of Europe. Contrary to other early Miocene herpetofaunas, the Echzell assemblage includes surprisingly only one form of crocodile-newts (Chelotriton). The Echzell Palaeobatrachus robustus represents the youngest record of the species and extends its stratigraphic range to the late early Miocene. Regarding chameleons, the frontal is partly preserved, but represents the first described frontal of the extinct species Chamaeleo andrusovi. The only anguine lizard that can be identified in the assemblage is represented by a new genus and species Smithosaurus echzellensis. Our phylogenetic analyses consistently recovered it as the sister taxon to either [Ophisauriscus quadrupes + Ophisaurus holeci] + [Anguis + Ophisaurus] (in the first analysis) or [Anguis + Ophisaurus] (in the second analysis). However, the results are based on limited fossil material – the parietal – and the support for the clade is very low. Thus, the interpretation of the Smithosaurus relationship among anguines needs to be taken with caution and has to be tested in further studies. Among snakes, Natrix longivertebrata represents the oldest record of the species and extends the stratigraphic range of this fossil snake back to the early Miocene. In addition, we provide here a broader comparison of the Echzell amphibian and reptilian assemblage with their European records for the MN3 and MN4 biostratigraphical units. Besides that, the entire herpetofauna of Echzell includes very broadly known early Miocene European forms. Remains of other groups of the same period such as Bufonidae, Hylidae, Pelodytidae, Amphisbaenia, Varanidae, Cordylidae, Pseudopus, are not found in the material available to us. We also conclude that the amphibian and reptilian fossil record across MN3–MN4 is significantly biased by taphonomic and/or environmental conditions. The amphibian and reptilian assemblage of Echzell is rich in forms living in humid and warm environments with forested areas, permanent water bodies and also some open habitats. The following climatic parameters can be reconstructed based on the herpetofauna: a mean annual temperature of 17.4–28.8 °C, minimal warm month temperature 18–28.3 °C, minimal cold month temperature 8–22.2 °C, and mean annual precipitation with a value of 791±254 mm.

Many animals interact with the air-water interface during locomotion. Such location either involves moving through the water's surface or moving atop the water surface. This dissertation aims to investigate both of these forms of locomotion in frogs. First we quantified the kinematics of skittering, jumping on top the water's surface without sinking, in two species of frog, Acris crepitans and Euphlyctis cyanophlyctis. We found that what was described as "skittering" locomotion in Acris crepitans is actually more akin to porpoising. A. crepitans begins and ends each jump during their interfacial behavior under the water surface. These frogs may be unable to perform true skittering locomotion due to not being able to retract their hindlimbs fast enough. E. cyanophlyctis, however, does stay above the water surface during this mode of locomotion. We found that Euphlyctis is highly maneuverable during skittering locomotion compared to other inertial based water-surface traversing animals. Not only can they turn up to 80° between subsequent jumps, they also perform this behavior in close proximity to each other without collision. Next, we investigated control mechanisms used by frogs when jumping from water. Prior research has identified frogs of the genus Euphlyctis as high jumpers. But previous studies only considered their maximal performance. Here, we investigated how these frogs modulate propulsive force in order to control their jump height. We linked the frog limb kinematics to the jump force by modeling the added mass produced by the foot's motion. Doctor of Philosophy There are many animals that move across or through the water's surface. Most of them are very small and light and can thus be supported by surface tension. Larger animals instead must produce the force needed to stay afloat by moving quickly. Previous research has looked at the physics involved in running on the water surface in basilisk lizards and grebes. However, the ability of frogs to jump on the water surface (a behavior known as "skittering") has never been studied. In this dissertation, we examine the water-surface traversal of two frog species, Acris crepitans and Euphlyctis spp., using high-speed videography. Unlike previous human observations in the literature, we found that Acris does not stay atop the water's surface during its jumping behavior and instead begins and ends each jump under the water. This is similar to porpoising in animals like dolphins. Euphlyctis, however, does stay above the water during this jumping behavior. We found that these frogs can turn sharply between jumps which has not been observed in any other large water-runners. Additionally, we studied the ability of Euphlyctis to jump high in the air starting from floating on the water surface. These frogs are interesting to study because they can jump unusually high compared to other species of similar size and shape. We found that when shown insects at different heights, these frogs can control their jump height and only jump as high as necessary. By tracking the frogs' limbs during jumping we investigated several possible ways these frogs controlled their jump height.

Amphibian skin contains wound-healing peptides, antimicrobial peptides, and insulin-releasing peptides, which give their skin a strong regeneration ability to adapt to a complex and harsh living environment. In the current research, a novel wound-healing promoting peptide, PM-7, was identified from the skin secretions of Polypedates megacephalus, which has an amino acid sequence of FLNWRRILFLKVVR and shares no structural similarity with any peptides described before. It displays the activity of promoting wound healing in mice. Moreover, PM-7 exhibits the function of enhancing proliferation and migration in HUVEC and HSF cells by affecting the MAPK signaling pathway. Considering its favorable traits as a novel peptide that significantly promotes wound healing, PM-7 can be a potential candidate in the development of novel wound-repairing drugs.

Imazethapyr is an herbicide that is used in a variety of crops worldwide, including soybean and corn. The aim of the present study was to evaluate the biomarkers responses of adult Leptodactylus latinasus exposed to the formulation Pivot® H (10.59% imazethapyr) in the laboratory at concentrations and under conditions that simulate two potential field exposure scenarios: an immersion in field runoff (Scenario 1: 10 mg/L) and a direct exposure to the droplets emitted by spray noozles (Scenario 2: 1000 mg/L). In both scenarios, the experimental procedure involved completely immersing the frogs over a period of 15 s. Different endpoints were evaluated at several ecotoxicological levels 48 and 96 h after the herbicide exposure. These included individual (biometric indices and behavior alterations), histological (liver pigments and lesions), biochemical (catalase, glutathione system and cholinesterase activities) and genotoxic effects (micronuclei induction and nuclear abnormalities). Forty-eight hours after imazethapyr exposure, frogs submitted to Scenario 1 presented an inhibition of liver glutathione-S-transferase activity, whereas histological alterations and increased hepatic cholinesterase levels were observed in frogs exposed under Scenario 2. Ninety-six hours after exposure to the imazethapyr formulation, frogs from the Scenario 1 treatment presented a decrease in liver melanin and hemosiderin, increased hepatic catalase activity and micronuclei induction. For their part, frogs exposed to Scenario 2 presented a decrease in the hepatosomatic index, an increase in liver alterations, melanin reduction and micronuclei induction. The multivariate analysis enables correlations to be made between biomarkers of different organizational level in exposed anurans. Our result indicates that real exposure to imazethapyr formulations under field conditions may pose a risk to Leptodactylus latinasus populations living in the agroecosystems., Facultad de Ciencias Naturales y Museo

Land use in Mexico has dramatically changed in recent decades since deforested lands have been repurposed for agriculture. We evaluated the amphibian taxonomic and functional diversity of a heterogeneous landscape with ten land cover/use types in west-central Mexico. Taxonomic diversity was evaluated with q-order indices, and functional diversity was calculated with three multivariate functional diversity indices by land cover/use. The relationship between amphibian diversity, habitat structure, and environmental variables was analyzed using multidimensional distance-based analyses. Our results showed that most native land cover types exhibited a similar species richness (low values) among the studied crops, except for the riparian habitat surrounded by tropical dry forest (high richness) and secondary vegetation (intermediate richness). Regarding functional diversity, the riparian habitat surrounded by tropical dry forest, sugar cane field, and secondary vegetation had the highest values. The secondary vegetation had more functional groups than other land cover/use types. Despite the lack of a clear spatial pattern of amphibian taxonomic and functional diversity, we determined that attributes such as herbaceous cover and water availability are essential to maintain both facets with high amphibian diversity in the land cover/use types (e.g., secondary vegetation and sugar cane).

People spend much of their time in imaginary worlds, and have beliefs about the events that are likely in those worlds, and the laws that govern them. Such beliefs are likely affected by people’s intuitive theories of the real world. In three studies, people judged the effort required to cast spells that cause physical violations. People ranked the actions of spells congruently with intuitive physics. For example, people judge that it requires more effort to conjure up a frog than to levitate it one foot off the ground. A second study manipulated the target and extent of the spells, and demonstrated with a continuous measure that people are sensitive to this manipulation even between participants. A pre-registered third study replicated the results of Study 2. These results suggest that people’s intuitive theories partly account for how they think about imaginary worlds.

Amid global biodiversity loss, it is important to find practical tools and solutions in order to protect biodiversity. Ecotourism is the fastest-growing sector of the international travel industry and can be a powerful conservation tool that encourages people to protect the natural environment. Traditionally, frogs have not generated much attention among ecotourists, partly because they are easily overshadowed by other more charismatic species or habitat attractions. With almost a third of the nearly 7000 known amphibian species listed as threatened by the International Union for Conservation of Nature (IUCN), their protection is crucial. Frogging is a well-known term within the frog conservation society, describing the activity of searching for frogs in the wild. This can be combined with other ecotourism activities to attract tourists and create an interest in the conservation of frogs while having fun at the same time. The aim was to determine the ecotourism potential of frogs in South Africa, primarily by distributing questionnaires to tourists to retrieve information on whether they would be interested in participating in frog-related ecotourism activities within the South African National Parks. For this research, a quantitative research approach was followed, namely non-probability sampling, to which convenience sampling was applied. An online survey (questionnaire) was designed to collect the data for the research. The survey outcome was satisfactory, as potential tourists indicated that they would like to participate in frog-related activities. The project offers the opportunity to conserve frogs, educate tourists, and create job opportunities within the local communities. It will also create a new tourism product for the South African National Parks.Conservation implications: The contribution of this research to conservation lies in the opportunity to benefit frog conservation through ecotourism.

Endemic anurans are particularly vulnerable to environmental changes, and are susceptible to population declines because of their restricted distribution ranges. The Murree Hills Frog Nanorana vicina and Hazara Torrent Frog Allopaa hazarensis are associated with the torrential streams and nearby clear water pools in subtropical chir pine forest and other forest types, at elevations higher than 1000 m in Pakistan. In this study, we have provided data on the extent of movement of these frog species for the first time. We installed radio transmitters on a total of 13 Murree Hills Frogs and 13 Hazara Torrent Frogs during eight consecutive days in September 2017 and 2018. Our results showed that these frogs did not move long distances along the stream or away from the stream into the forest. All the radio-tracked frogs showed movement of < 3 m. We found a significant differences only in the distance moved by Murree Hills Frogs between the two years studied. Based on our findings, we propose a movement paradigm that focuses on conservation implications for these endemic frogs.

Reptiles and amphibians are experiencing declines across the globe. In Monteverde, Costa Rica, these declines and their underlying causes have been relatively well studied since the early 1990s, and many protected areas have been set aside to conserve these species. However, thorough surveys of the herpetofaunal diversity in these areas have been scarce over the last 20 years. We conducted a survey of all reptile and amphibian species at the University of Georgia Costa Rica (UGACR), a field station in San Luis de Monteverde. Herein, we present an annotated checklist of the 48 species (35 reptiles and 13 amphibians) that we encountered. While we did not find any exceptionally rare or endangered species, the number of species we encountered is disproportionately high given the small plot of land occupied by UGACR. This underscores the importance of conducting regular diversity surveys in biodiversity hotspots as a means to better inform conservation efforts.

An annotated inventory of the herpetofauna of Lawachara National Park in Bangladesh is presented, based primarily on original field observations recorded during a six-year survey of the park. A total of 71 species are reported, including 19 Anura, one Apoda, two Chelonii, and 49 Squamata. The course of the survey revealed 16 range extensions including 11 new country records for Bangladesh. Eight of the 16 range extensions including six of the 11 country records are reported here for the first time. Deleted from previous Lawachara National Park checklists are 23 species that had been erroneously reported due to misidentification, a species split, or the subsumption of the species within another taxa.

Made available in DSpace on 2020-12-12T02:22:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 In recent years, the taxonomic knowledge of Pseudopaludicola has increased and it is currently possible to elucidate aspects of the species within this genus such as their geographic distributions. Here we accessed new data on geographic distribution of P. boliviana and three long-legged species of the P. saltica group (P. saltica, P. murundu, and P. jaredi). Based on morphometric, acoustic and genetic data, we accessed the specific status of long-legged specimens from Chapada dos Veadeiros highlands and attributed them to the P. murundu. We also performed a phylogenetic analysis including several populations of the three species. Based on this novel dataset, we discussed the geographical distributions of P. saltica, P. murundu, and P. jaredi, providing new records for them. Pseudopaludicola boliviana is one of the five species of the P. pusilla group and it is thought to be a widely distributed species based on its morphology. Hence, we evaluated the geographic distribution of P. boliviana, using novel acoustic data of specimens from Argentina, Northern Brazil, and Bolivia. Laboratório de História Natural de Anfíbios Brasileiros (LaHNAB) Departamento de Biologia Animal Instituto de Biologia Universidade Estadual de Campinas (UNICAMP Laboratório de Taxonomia e Sistemática de Anuros Neotropicais (LTSAN) Instituto de Ciências Exatas e Naturais do Pontal Universidade Federal de Uberlândia (UFU) Instituto de Biologia Universidade Estadual de Campinas (UNICAMP Laboratório de Herpetologia Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP, Rio Claro Laboratório de Citogenética Animal Departamento de Biologia Instituto de Biociências Universidade Estadual Paulista (UNESP, Rio Claro Laboratorio de Herpetología Universidad Nacional del Nordeste Laboratório de Herpetologia Departamento de Biodiversidade e Centro de Aquicultura (CAUNESP) Instituto de Biociências Universidade Estadual Paulista (UNESP, Rio Claro Laboratório de Citogenética Animal Departamento de Biologia Instituto de Biociências Universidade Estadual Paulista (UNESP, Rio Claro

The recent detection of a novel amoebozoan parasite (Entamoeba sp. CT1) killing invasive cane toads (Rhinella marina) in tropical Australia raises concerns of potential spill-over into native anuran populations. Considering the vulnerability of anuran communities globally, Entamoeba sp. CT1 may pose a serious threat to anuran biodiversity. Through PCR-based detection and molecular identification, we investigated the prevalence of Entamoeba spp. in the faeces and colon tissue of cane toads (Rhinella marina) and eleven native Australian frog species from a single locality in the Northern Territory. No Entamoeba DNA was detected in samples of native frog faeces (N = 57) or colons (N = 17). Entamoeba DNA was detected in 24% of 45 cane toads (95%CI 14.08–38.82). Both E. ranarum and Entamoeba sp. CT1 were present in cane toads. The failure of faecal samples to indicate Entamoeba spp. in infected cane toads may be due to cysts in faeces being shed intermittently, degraded before analysis, or impervious to lysis prior to DNA isolation. Our results suggest that native frogs do not carry the pathogen in an area where 20–30% of cane toads are infected with Entamoeba sp. CT1. We demonstrate the importance of recognising PCR inhibition prior to molecular diagnostics, and the apparent inadequacy of faecal samples for the detection of Entamoeba spp. in anurans., Graphical abstract Image 1, Highlights • Entamoeba spp. were detected in cane toads, but not in faeces or colons of native frogs. • PCR inhibition of anuran faecal samples. • Additional purification, the presence of inhibition was reduced.

Mexico City houses one of the most populous urban areas of the world, and the modification of its natural habitat likely influences the biological diversity found there. In particular, amphibians and reptiles are likely affected by these modifications. Herein, we present an updated list of the species of amphibians and reptiles that inhabit Mexico City. Mexico City harbors 65 species of amphibians and reptiles, which represent 21 families and 33 genera. These include 18 species of amphibians (nine anurans and nine salamanders) and 47 species of reptiles (14 lizards, 30 snakes [one introduced], and three turtles [one introduced]). Forty-eight of the amphibian and reptile species in Mexico City are endemic to Mexico, with two endemic to Mexico City. The most diverse region of Mexico City is the Forests and Ravines region, which is home to 43 species. Eleven species of amphibians and reptiles in Mexico City are IUCN listed, 16 are placed in a protected category by SEMARNAT (Secretaria del Medio Ambiente y Recursos Naturales), and 27 species are categorized as high risk by the EVS (Environmental Viability Score). Mexico City shares almost 94% of its species with the State of Mexico.

Despite being one of the smallest states in Mexico, the high diversity of habitats in Morelos has led to the development of a rich biota made up of a mixture of species typical of the Neovolcanic Axis and the Sierra Madre del Sur. However, recent expansion of cities in Morelos is likely to have consequences for the state’s herpetofauna. Here a checklist of the amphibians and reptiles of Morelos is provided with a summary of their conservation status and overlap with its neighboring states. Morelos is home to 139 species of amphibians and reptiles representing 32 families and 75 genera. Twenty-six of the 38 species of amphibians and 70 of the 101 species of reptiles that inhabit Morelos are endemic to Mexico. Fourteen species of amphibians and reptiles from Morelos are IUCN listed (i.e., Vulnerable, Near Threatened, or Endangered), 22 are placed in a protected category by SEMARNAT, and 41 are categorized as high risk by the EVS. The Tropical Deciduous Forest vegetation type hosts the greatest number of amphibian and reptile species in Morelos (84 species). Morelos shares the largest proportion of its herpetofauna with the State of Mexico (79.3%), Puebla (77.0%), and Guerrero (74.8%).

A contemporary review of the studies on the trophic spectrum and feeding ecology of the Bulgarian frogs and toads is presented. The analysis of the Bulgarian herpetological literature showed that currently, there are 18 specialized studies on the diet of the Bulgarian anuran species. Of all 16 anuran species, occurring in Bulgaria, 9 (56.25%) have been studied in connection with their trophic spectrum. For four species (25.00%), there is very little data, and for 7 (43.75%), such studies have not yet been done. All Bulgarian species of amphibians are zoophagous, mainly insectivorous. All studied species are general feeders (polyphages), except Bombina variegata, which shows a slight preference to Coleoptera and Bufo bufo and Bufotes viridis, which show slight preference to Formicidae and Coleoptera. Cannibalism was recorded only in Pelophylax ridibundus. The highest values of the trophic niches breadths are recorded in P. ridibundus, R. graeca, R. temporaria, and R. dalmatina. The trophic niches breadths of the other anuran species found in Bulgaria have significantly lower values, the lowest being in Hyla orientalis.

The ultrastructural characteristics of the mature spermatozoon of Diplodiscus amphichrus (Digenea, Paramphistomoidea, Diplodiscidae) and their ultrastructural organisation were examined by means of transmission electron microscopy. Live digeneans were collected from the Chinese edible frog Hoplobatrachus rugulosus in Thailand. The male gamete of D. amphichrus is a filiform cell with two axonemes of the 9 + ’1’ trepaxonematan pattern, nucleus, one mitochondrion, parallel cortical microtubules, a well-developed lateral expansion, external ornamentation of the plasma membrane, spine-like bodies, and granules of glycogen. These ultrastructural characteristics have already been described in other paramphistomoids studied so far except for the cladorchiids, which present some differences. Two characteristics of the male gamete of D. amphichrus were found for the first time in a digenean: (i) the appearance of the initial part of the lateral expansion before the axonemes in the anterior extremity and (ii) the separation between the second axoneme and the nucleus in the posterior tip. Our results are compared with the available data in the Digenea and particularly with other paramphistomoids.

Eupsophus migueli is considered a microendemic endangered species inhabiting the temperate Nothofagus forests of the Mahuidanche Range of southern Chile. However, this categorization is based on scarce data about its distribution and natural history. In order to assess these parameters, this article reports new geographic records obtained through intensive fieldwork between 2011 and 2016. Considering this, an updated distribution map for E. migueli is proposed, and new data about natural history traits and habitat use are provided. The information obtained in this study is discussed considering the zoogeographical importance of E. migueli, and confirms the species IUCN conservation status.

Colima is the fourth smallest Mexican state, covering only 0.3% of the surface area of Mexico, but due to the remarkable diversity of physiographic and environmental conditions present in Colima it contains a high biological diversity. We generated an up-to-date herpetofaunal checklist for Colima, with a summary of the conservation status of Colima’s amphibians and reptiles. Our checklist contains a total of 153 species of amphibians and reptiles (three introduced). Thirty-nine are amphibians and 114 are reptiles. More than half of Colima’s herpetofauna are Mexican endemics (66.7% of amphibians, 67.5% of reptiles). Less than 25% of the amphibian and reptile species in Colima are in protected categories according to the IUCN Red List and SEMARNAT. The reptiles in the Marine and Revillagigedo Archipelago regions are the most threatened taxa of the Colima herpetofauna. Colima shares > 80% of its herpetofauna with its neighboring states, Jalisco and Michoacán.

Haemogregarine (Apicomplexa: Adeleorina) blood parasites are commonly reported from anuran hosts. Dactylosomatidae (Jakowska and Nigrelli, 1955) is a group of haemogregarines comprising Dactylosoma Labbé, 1894 and Babesiosoma Jakowska and Nigrelli, 1956. Currently Dactylosoma and Babesiosoma contain five re-cognised species each. In the current study, a total of 643 anurans, comprising 38 species, 20 genera, and 13 families were collected from South Africa (n = 618) and Belgium (n = 25), and their blood screened for the presence of dactylosomatid parasites. Three anuran species were found infected namely, Ptychadena anchietae (Bocage, 1868) and Sclerophrys gutturalis (Power, 1927) from South Africa, and Pelophylax lessonae (Camerano, 1882) from Belgium. Based on morphological characteristics, morphometrics and molecular results a new dactylosomatid, Dactylosoma kermiti n. sp. is described form Pty. anchietae and Scl. gutturalis. The species of Dactylosoma isolated from Pel. lessonae could not, based on morphological or molecular analysis, be identified to species level. Phylogenetic analysis shows species of Dactylosoma infecting anurans as a monophyletic group separate from the other haemogregarine groups. Additionally, the mosquitoes Uranotaenia (Pseudoficalbia) ma-shonaensis Theobald, 1901 and U. (Pfc.) montana Ingram and De Meillon, 1927 were observed feeding on Scl. gutturalis in situ and possible dividing stages of this new parasite were observed in the mosquitoes. This study is the first to describe a dactylosomatid parasite based on morphological and molecular data from Africa as well as observe potential stages in possible dipteran vectors. This paper forms part of a VLIR-UOS TEAM funded project (ZEIN21013PR396), co-funded by the National ResearchFoundation of South Africa (NRF project CPRR160429163437, grant105979, NJ Smit, PI). The financial assistance of the NRF towards ECNsupported by the DAAD-NRF doctoral scholarship (Grant UID: 108803), and the VLIR-OUS university scholarship (ID 0620854/Contract000000076310), is acknowledged

Summary Two species of digenetic trematodes of the genus Pleurogenoides viz., P. cyanophlyctiShinad & Prasadan (2018a) and P. euphlyctiShinad & Prasadan (2018b) have been described from India. Information regarding the molecular data of various species of the genus Pleurogenoides Travassos, 1921 is virtually lacking. This study addresses the application of molecular markers to validate the phylogenetic position of P. cyanophlycti and P. euphlycti. In the present study, two species P. cyanophlycti and P. euphlycti were collected between January 2016 to October 2017, infecting the freshwater frogs inhabiting the Western Ghats, India. In the present study, the two species were identifi ed morphologically and by PCR amplification of the 28S ribosomal RNA gene. Phylogenetic tree results clearly demonstrate that both P. cyanophlycti and P. euphlycti belongs to the family Pleurogenidae Looss, 1899. Based on these results, we presented and discussed the phylogenetic relationships of P. cyanophlycti and P. euphlycti within family Pleurogenidae from India. Phylogenetic analyses showed that P. cyanophlycti and P. euphlycti cluster according to their vertebrate host and revealed an important congruence between the phylogenetic trees of Pleurogenoides and of their vertebrate hosts. P. cyanophlycti and P. euphlycti clearly constitute a separate, sister branch with other species of the genera, Pleurogenoides, Pleurogenes (=Candidotrema), Prosotocus and Brandesia. The present study firstly provides important information about the molecular study and phylogenetic analysis of P. cyanophlycti and P. euphlycti. This study will also serve as a baseline for Pleurogenoides species identifi cation for further studies.

Given its topicality, it is tempting to suppose that one may find important insights into the politics of late 5th C. Athens in Aristophanes’ comedies. The problem, I contend, is when scholars think they can discern Aristophanes’ own political views simply by supposing that some character in the play (or the chorus leader in the parabasis) directly presents the author’s views. As tempting as such an inference sometimes is, it is one that should be made with extreme caution. For each example of what might seem to some scholars as serious political advice, one may find many other instances that cannot possibly be taken to represent Aristophanes’ real views in the lines he has written. In this discussion, I take up just one case of political speech in an Aristophanic play, Frogs, and argue (contrary to most existing scholarship) that it should not be interpreted as didacticism. Instead, I argue that Aristophanes gives samples of political advocacy from the most extreme poles of contemporary ideology, in such a way as to highlight how dangerous and foolish such policies would be. Aristophanes was mocking, not endorsing, the follies that would soon prove to be so ruinous for Athens.

Different frog-bromeliad associations are frequently reported from various habitats of Central and South America. Here, we present the first data on the association of two hylid species (Dendropsophus nanus and Scinax nasicus) with the bromeliad Bromelia balansae in the Brazilian Pantanal. Both treefrog species use rosettes of B. balansae as a diurnal shelter and foraging ground. Made available in DSpace on 2020-09-02T04:38:51Z (GMT). No. of bitstreams: 1 AnuranBromeliaBalansaes2020.pdf: 1995547 bytes, checksum: 57925c299270528867b44189a6311058 (MD5) Previous issue date: 2020

The behavioral study is essential to improve the understanding of the natural history of species, especially when it involves predatory organisms that are essential parts in the structuring of biological communities. In the case of anuran amphibians, there are already reports of some species that tend to present generalist and opportunistic behaviors, with the amphibians of the Bufonidae family being good representatives to demonstrate behaviors of this character. Therefore, the objective of this communication was to describe an unprecedented event involving the scavenging act for the species R. diptycha in an urban environment, which is an unusual behavior for anuran amphibians. Along with the report, a compilation of events involving predator/prey interactions with amphibians was carried out, making it possible to evidence the Bufonidae family as the most representative among those that were sampled. El estudio del comportamiento es esencial para mejorar la comprensión de la historia natural de las especies, especialmente cuando se trata de organismos depredadores que son partes esenciales en la estructuración de las comunidades biológicas. En el caso de los anfibios anuros, ya existen reportes de algunas especies que tienden a presentar comportamientos generalistas y oportunistas, siendo los anfibios de la familia Bufonidae buenos representantes para demostrar comportamientos de este carácter. Por lo tanto, el objetivo de esta comunicación fue describir un evento sin precedentes que involucra el acto de carroñeo de la especie R. diptycha en un ambiente urbano, lo cual es un comportamiento inusual para los anfibios anuros. Junto con el informe se realizó una recopilación de eventos de interacciones depredador/presa con anfibios, lo que permitió evidenciar a la familia Bufonidae como la más representativa entre las muestreadas. O estudo comportamental é fundamental para melhorar a compreensão da história natural das espécies, principalmente quando envolve organismos predadores que são peças essenciais na estruturação de comunidades biológicas. Tratando-se dos anfíbios anuros, já se tem relatos de algumas espécies que tendem a apresentar comportamentos generalistas e oportunistas, sendo os anfíbios da família Bufonidae bons representantes para evidenciar comportamentos desse caráter. Portanto, objetivou-se com a presente comunicação descrever um evento inédito envolvendo o ato necrófago para a espécie R. diptycha em ambiente urbano, sendo este um comportamento incomum para anfíbios anuros. Juntamente ao relato, foi realizada uma compilação de eventos envolvendo interações predador/presa com anfíbios, sendo possível evidenciar a família Bufonidae como a mais representativa entre as que foram amostradas.

Ribosomes are essential nanomachines responsible for protein production. Although ribosomes are present in every living cell, ribosome biogenesis dysfunction diseases, called ribosomopathies, impact particular tissues specifically. Here, we evaluate the importance of the box C/D snoRNA-associated ribosomal RNA methyltransferase fibrillarin (Fbl) in the early embryonic development of Xenopus laevis. We report that in developing embryos, the neural plate, neural crest cells (NCCs), and NCC derivatives are rich in fbl transcripts. Fbl knockdown leads to striking morphological defects affecting the eyes and craniofacial skeleton, due to lack of NCC survival caused by massive p53-dependent apoptosis. Fbl is required for efficient pre-rRNA processing and 18S rRNA production, which explains the early developmental defects. Using RiboMethSeq, we systematically reinvestigated ribosomal RNA 2’-O methylation in X. laevis, confirming all 89 previously mapped sites and identifying 15 novel putative positions in 18S and 28S rRNA. Twenty-three positions, including 10 of the new ones, were validated orthogonally by low dNTP primer extension. Bioinformatic screening of the X. laevis transcriptome revealed candidate box C/D snoRNAs for all methylated positions. Mapping of 2’-O methylation at six developmental stages in individual embryos indicated a trend towards reduced methylation at specific positions during development. We conclude that fibrillarin knockdown in early Xenopus embryos causes reduced production of functional ribosomal subunits, thus impairing NCC formation and migration., Author summary Ribosomes are essential nanomachines responsible for protein production in all cells. Ribosomopathies are diseases caused by improper ribosome formation due to mutations in ribosomal proteins or ribosome assembly factors. Such diseases primarily affect the brain and blood, and it is unclear how malfunctioning of a process as general as ribosome formation can lead to tissue-specific diseases. Here we have examined how fibrillarin, an enzyme which modifies ribosomal RNA by adding methyl groups at specific sites, affects early embryonic development in the frog Xenopus laevis. We have revealed its importance in the maturation of cells forming an embryonic structure called the neural crest. Fibrillarin depletion leads to reduced eye size and abnormal head shape, reminiscent of other conditions such as Treacher Collins syndrome. Molecularly, the observed phenotypes are explainable by increased p53-dependent programmed cell death triggered by inhibition of certain pre-rRNA processing steps. Our systematic investigation of the ribosomal RNA 2’-O methylation repertoire across development has further revealed hypomodification at a late stage of development, which might play a role in late developmental transitions involving differential translation by compositionally different ribosomes.

This study could not have been possible without the team that uncovered this specimen, Farish A Jenkins Jr., William W. Amaral, William R. Downs III, Stephen M. Gatesy, Neil H. Shubin Niels Bonde and Lars B. Clemmensen. We thank Harvard University and Bent Lindow from the Natural History Museum of Denmark. Thanks to Alexandre Guillaume and Vincent Cheng for reviewing and bringing improvements to the original manuscript and Carla Tomás for laboratory support. Alfred Lemierre and an anonymous reviewer are thanked for their comments on the manuscript. During the Triassic, Batrachia diverged into ancestors of frogs (Salientia) and salamanders (Caudata). Fossils of Triassic batrachians are rare and found only in a few outcrops, such as the Middle Sakamena Formation of Madagascar (Induan). Only three Triassic taxa have been described, the two early frogs Triadobatrachus and Czatkiobatrachus and the early salamander Triassurus. Here we describe a right ilium, collected in 1991, attributed to the first batrachian from the Late Triassic Carlsberg Fjord Member (Ørsted Dal Formation, Fleming Fjord Group) in the Jameson Land Basin, located in central East Greenland. The fossil specimen only displays the proximal part of a right ilium, missing its shaft. After a thorough comparison with several clades (lizards, temnospondyls, salamanders and frogs), we consider the specimen as a lissamphibian sharing feature with salientians and anurans: squarish acetabular region, deeply concave acetabular surface, laterally projecting acetabular rim, flat mesial surface. It is the youngest Triassic specimen of Batrachia to date and one of the northernmost of the Late Triassic. publishersversion publishersversion published

Thesis (Masters by research (Medical Science)) --University of South Australia, 2022. Includes bibliographical references (pages 129-134) The spread of the frog killing Batrachochytrium dendrobatidis (Bd) fungus is contributing to the world’s sixth mass extinction, with frog populations undergoing unprecedented declines. This thesis describes the isolation of microorganisms that have the ability to inhibit the growth of this pathogen. The study involved the isolation of microorganisms from the skin of native frog species and a subsequent series of in-vitro challenge assays. It also included a frog captivity study to determine how the loss of the frogs’ natural environment and changes in water salinity effect the frog skin microbiota. Two probiotic candidates were identified from the studies for future probiotic trials. This body of work is the first step in the development of an effective bioaugmentation strategy to combat the spread of Bd in South Australia.

We present a list of amphibian and reptile species encountered from 16–20 July 2016 in the upper drainage of Río Guázaro, Parque Nacional Santa Fé, Veraguas, Panama. We collected 10 amphibian and 10 reptile species and observed three additional amphibian and one additional reptile species. Our collected material provides the first records from Veraguas province for the amphibians Bolitoglossa schizodactyla Wake & Brame, 1966 and Diasporus aff. quidditus (Lynch, 2001), as well as for the reptiles Basiliscus plumifrons Cope, 1876, Micrurus stewarti Barbour & Amaral, 1928, and Sibon argus (Cope, 1876). Although relatively small in species and specimen numbers, our collection represents the first documented herpetological inventory on the largely unexplored Caribbean versant of Veraguas in 50 years.

Emerging infectious diseases are increasing globally and are an additional challenge to species dealing with native parasites and pathogens. Therefore, understanding the combined effects of infectious agents on hosts is important for species’ conservation and population management. Amphibians are hosts to many parasites and pathogens, including endemic trematode flatworms (e.g., Echinostoma spp.) and the novel pathogenic amphibian chytrid fungus (Batrachochytrium dendrobatidis [Bd]). Our study examined how exposure to trematodes during larval development influenced the consequences of Bd pathogen exposure through critical life events. We found that prior exposure to trematode parasites negatively impacted metamorphosis but did not influence the effect of Bd infection on terrestrial growth and survival. Bd infection alone, however, resulted in significant mortality during overwintering—an annual occurrence for most temperate amphibians. The results of our study indicated overwintering mortality from Bd could provide an explanation for enigmatic declines and highlights the importance of examining the long-term consequences of novel parasite exposure.

Sri Lanka is an amphibian hotspot of global significance. Its anuran fauna is dominated by the shrub frogs of the genusPseudophilautus. Except for one small clade of four species in Peninsular India, these cool-wet adapted frogs, numbering some 59 extant species, are distributed mainly across the montane and lowland rain forests of the island. With species described primarily by morphological means, the diversification has never yet been subjected to a molecular species delimitation analysis, a procedure now routinely applied in taxonomy. Here we test the species boundaries ofPseudophilautusin the context of the phylogenetic species concept (PSC). We use all the putative species for which credible molecular data are available (nDNA–Rag-1; mt-DNA– 12S rRNA, 16S rRNA) to build a well resolved phylogeny, which is subjected to species delimitation analyses. The ABGD, bPTP, mPTP and bGMYC species delimitation methods applied to the 16S rRNA frog barcoding gene (for all species), 12S rRNA and Rag-1 nDNA groupedP.procaxandP.abundus;P.hallidayiandP.fergusonianus;P.reticulatusandP.pappilosus;P.pleurotaeniaandP.hoipolloi;P.hoffmaniandP.asankai;P.silvaticusandP.limbus;P.dilmahandP.hankeni;P.fulvusandP.silus.. Surprisingly, all analyses recovered 14 unidentified potential new species as well. The geophylogeny affirms a distribution across the island’s aseasonal ‘wet zone’ and its three principal hill ranges, suggestive of allopatric speciation playing a dominant role, especially between mountain masses. Among the species that are merged by the delimitation analyses, a pattern leading towards a model of parapatric speciation emerges–ongoing speciation in the presence of gene flow. This delimitation analysis reinforces the species hypotheses, paving the way to a reasonable understanding of Sri LankanPseudophilautus, enabling both deeper analyses and conservation efforts of this remarkable diversification.http://zoobank.org/urn:lsid:zoobank.org:pub:DA869B6B-870A-4ED3-BF5D-5AA3F69DDD27.

Since the establishment of the Nam Xuan Lac Habitat and Species Conservation Area in 2003 in Bac Kan Province, northern Vietnam, only two herpetological studies have been conducted: One recorded 14 species of amphibians from Ban Thi-Xuan Lac area in 2004 and a recent study reported 32 species of amphibians from this protected area in 2019. As a result of our field surveys in 2020 and 2021, a total of 23 species of amphibians was recorded from the Nam Xuan Lac Habitat and Species Conservation Area. Eight of them are recorded for the first time from Bac Kan Province, comprising one species of Microhylidae, two species of Megophryidae, one species of Dicroglossidae, two species of Ranidae and two species of Rhacophoridae. Besides morphological descriptions, we provide ecological notes of newly-recorded species of amphibians from Bac Kan Province.

To determine trends in either frog distribution or abundance in the State of Louisiana, we reviewed and analyzed frog call data from the Louisiana Amphibian Monitoring Program (LAMP). The data were collected between 1997 and 2017 using North American Amphibian Monitoring Program protocols. Louisiana was divided into three survey regions for administration and analysis: the Florida Parishes, and 2 areas west of the Florida parishes called North and South. Fifty-four routes were surveyed with over 12,792 stops and 1,066 hours of observation. Observers heard 26 species of the 31 species reported to be in Louisiana. Three of the species not heard were natives with ranges that did not overlap with survey routes. The other two species were introduced species, the Rio Grande Chirping Frog (Eleutherodactylus cystignathoides) and the Cuban Treefrog (Osteopilus septentrionalis). Both seem to be limited to urban areas with little to no route coverage. The 15 most commonly occurring species were examined in detail using the percentage of stops at which they observed along a given survey and their call indices. Most species exhibited a multimodal, concave, or convex pattern of abundance over a 15-year period. Among LAMP survey regions, none of the species had synchronous population trends. Only one group of species, winter callers, regularly co-occur. Based on the species lists, the North region could be seen as a subset of the South. However, based on relative abundance, the North was more similar to Florida parishes for both the winter and summer survey runs. Our analyses demonstrate that long-term monitoring (10 years or more) may be necessary to determine population and occupancy trends, and that frog species may have different local demographic patterns across large geographic areas.

Os nanomateriais (NMs) têm sido usados em um número crescente de produtos comerciais e essa rápida expansão pode levar à sua liberação em ambientes aquáticos. Atualmente a acelerada produção dos diversos tipos de produtos convencionais em nanoescala tem sido fundamental para o progresso econômico de vários países. Muitos tipos de nanopartículas e nanomateriais já estão sendo utilizados e outras variedades estão previstas para aparecer no futuro, no entanto, é escasso o conhecimento sobre o impacto dos NMs na biota, principalmente nos anfíbios. Sendo assim, há a necessidade de classificar e compreender melhor os NMs. A primeira parte de nosso estudo revisou o uso histórico de espécies de anfíbios como sistemas modelo em estudos nanoecotoxicológicos, no qual resumimos os dados disponíveis na literatura científica sobre os efeitos genotóxicos, mutagênicos, histopatológicos, embriotóxicos e reprodutivos de NMs em diferentes grupos de anfíbios. A interação, bioacumulação, modo de ação (MoA) e ecotoxicidade de NMs em anfíbios também foram revisados. Os estudos nanoecotoxicológicos foram realizados com 12 espécies de anfíbios, sendo oito da ordem Anura e três da ordem Caudata. Xenopus laevis foi a espécie mais estudada. Os estudos foram conduzidos principalmente com NMs inorgânicos (72%) em comparação com os orgânicos. A nanoecotoxicidade depende do comportamento do NM e da transformação em meio ambiente, bem como dos estágios de desenvolvimento dos anfíbios. O MoA de NMs em anfíbios foi associado principalmente à produção de espécies reativas de oxigênio (ROS), estresse oxidativo, efeitos genotóxicos e mutagênicos. Os resultados enfatizaram a necessidade de mais estudos que testem a ecotoxicidade de diferentes NMs, concentrações e períodos de exposição em abordagens ambientalmente relevantes. Além disso, há a necessidade de protocolos padrão para testes nanoecotoxicológicos com anfíbios. Os dados revisados mostraram que os anfíbios são organismos adequados para avaliar o impacto ambiental dos NMs e indicaram importantes lacunas na pesquisa sobre a ecotoxicidade dos NMs em ecossistemas de água doce e recomendações para pesquisas futuras. Nesse sentido, observamos a partir do primeiro capítulo que entre as nanopartículas mais produzidas e consumidas mundialmente estão as nanopartículas de dióxido de titânio (NPs de TiO2), cuja liberação no meio ambiente pode induzir efeitos tóxicos já reconhecidos em invertebrados aquáticos e vertebrados. No entanto, o conhecimento de seu impacto sobre os anfíbios neotropicais permanece limitado. Assim, o segundo capítulo teve como objetivo avaliar a toxicidade de NPs de TiO2 e sua contraparte dissolvida, dióxido de titânio (TiO2), em uma espécie de anfíbio anuro neotropical (Dendropsophus minutus). Danos ao DNA, parâmetros biométricos e mudanças comportamentais foram analisados em girinos após exposição a três concentrações ambientalmente relevantes (0,1, 1,0 e 10 mg L-1) de NPs de TiO2 e TiO2 por 7 dias. Como resultado, verificamos danos ao DNA em girinos de D. minutus expostos a ambas as formas de Ti em comparação ao grupo controle. Também identificamos uma redução semelhante no tamanho total, comprimento do corpo, largura e altura da musculatura da cauda em girinos expostos a NPs de TiO2 e TiO2 em comparação com girinos não expostos. Em relação ao teste comportamental, os girinos apresentaram menor mobilidade e ficaram mais distantes dos coespecíficos (menos agregados) quando expostos aos NPs. Portanto, o uso simultâneo de múltiplos biomarcadores foi essencial para avaliar os efeitos adversos do nanomaterial e estabelecer uma abordagem confiável para o biomonitoramento de ecossistemas aquáticos. Nosso estudo aumenta o conhecimento sobre os efeitos genotóxicos, morfológicos e comportamentais dos NPs de TiO2 e TiO2 em anfíbios anuros, contribuindo para estudos futuros na avaliação de risco ambiental de nanomateriais. Nanomaterials (NMs) have been used in an increasing number of commercial products, and its rapid expansion could lead to their release into aquatic environments. The accelerated production of the various types of conventional products at nanoscale has been fundamental for the economic progress of several countries. Many types of nanoparticles and nanomaterials are already being used, and other varieties are expected to appear in the future. However, there is little knowledge about the impact of NMs on biota, especially on amphibians. Therefore, there is a need to better classify and understand NMs. The first part of our study has reviewed the historical use of amphibian species as model systems in nanoecotoxicological studies, which summarized data available in the scientific literature on the genotoxic, mutagenic, histopathological, embryotoxic, and reproductive effects of NMs in different groups of amphibians. The interaction, bioaccumulation, mode of action (MoA), and ecotoxicity of NMs in amphibians were also reviewed. The nanoecotoxicological studies were carried out with 12 species of amphibians, eight from the Anura order and three from the Caudata order. Xenopus laevis was the most studied species. Studies were conducted primarily with inorganic NMs (72%) compared to organic ones. Nanoecotoxicity depends on the behavior of the NM, the transformation in the environment, and the developmental stages of the amphibians. The MoA of NMs in amphibians was mainly associated with the production of reactive oxygen species (ROS), oxidative stress, genotoxic and mutagenic effects. The results emphasized the need for more studies that test the ecotoxicity of different NMs, concentrations, and exposure periods in environmentally relevant approaches. In addition, there is a need for standard protocols for nanoecotoxicological testing with amphibians. The reviewed data showed that amphibians are suitable organisms to assess the environmental impact of NMs and indicated essential gaps in research on the ecotoxicity of NMs in freshwater ecosystems, as well as recommendations for future research. In this sense, we observed from the first chapter that the most produced and consumed nanoparticles worldwide are titanium dioxide nanoparticles (TiO2 NPs); whose release into the environment are known to induce toxic effects in aquatic invertebrates and vertebrates. However, knowledge of its impact on neotropical amphibians remains limited. Thus, the second chapter of this thesis aimed to evaluate the toxicity of TiO2 NPs and its dissolved counterpart, titanium dioxide (TiO2), in a species of anuran neotropical amphibian (Dendropsophus minutus). DNA damage, biometric parameters, and behavioral changes were analyzed in tadpoles after exposure to three environmentally relevant concentrations (0.1, 1.0, and 10 mg L-1) of TiO2 and TiO2 NPs for 7 days. As a result, we verified DNA damage in D. minutus tadpoles exposed to both forms of Ti compared to the control group. We also identified a similar reduction in overall size, body length, width, and height of the tail musculature in tadpoles exposed to TiO2 and TiO2 NPs compared to unexposed tadpoles. Regarding the behavioral test, the tadpoles showed less mobility and were more distant from the conspecifics (less aggregated) when exposed to NPs. Therefore, the simultaneous use of multiple biomarkers was essential to assess the adverse effects of the nanomaterial and establish a reliable approach for biomonitoring of aquatic ecosystems. Our study increases the knowledge about the genotoxic, morphological, and behavioral effects of TiO2 and TiO2 NPs in anuran amphibians, contributing to future studies in the environmental risk assessment of nanomaterials.

Coronaviruses are pathogens recognized for having an animal origin, commonly associated with terrestrial environments. However, in a few cases, there are reports of their presence in aquatic organisms like fish, frogs, waterfowl, and marine mammals. None of these cases has led to human health effects when contact with these infected organisms has taken place, whether they were alive or dead. Aquatic birds seem to be the main group carrying and circulating these types of viruses among healthy bird populations. Although the route of infection for COVID-19 by water or aquatic organisms has not yet been observed in the wild, the relevance of its study is highlighted because there are cases of other viral infections known to have been transferred to humans by aquatic biota. It is encouraging to know that aquatic species, such as fish, marine mammals, and amphibians, show very few coronavirus cases. Some other aquatic animals may also be a possible source of cure or treatment against, as some evidence with algae and aquatic invertebrates suggest.

In tropical regions presenting severe seasonality, the reproductive activity of many species occurs during the most appropriate periods of the year. Therefore, several physiological and behavioral adjustments are needed during their reproductive cycle. This study aims to evaluate possible changes in histologic and morphometric parameters of the ovaries as well in estrogen and progesterone plasma levels of Leptodactylus macrosternum females and their relationship with temperature and precipitation. Specimens were collected in the Horto Florestal Olho d’Água da Bica - HFOB (06°49´20’’S/36°15´85’’W) area in the municipality of Cuité, brazilian semiarid, every 15 days, only between the months of May to August and November to December. The quantification of population density of oocyte types and dosages of progesterone and estrogen levels were used to determine the reproductive activity of the species. Population densities of oogonia and oocyte II did not vary significantly during the months in which the animals were sampled. Population densities of oocyte I was significantly higher during the months of June (p= 0.04) and August (p= 0.03). The highest values for the population density of oocyte III and IV were found in the months of May (p=0.04) and June (p=0.04). The amounts of estrogen concentrations (pg / ml) and progesterone (pg / ml) did not change significantly over the sampling periods. The population density of the types of oocytes III and IV showed dependence on the variation in rainfall. En las regiones tropicales que presentan una estacionalidad severa, la actividad reproductiva de muchas especies ocurre durante los períodos más apropiados del año. Por lo tanto, se necesitan varios ajustes fisiológicos y de comportamiento durante su ciclo reproductivo. Este estudio tiene como objetivo evaluar los posibles cambios en los parámetros histológicos y morfométricos de los ovarios, así como en los niveles plasmáticos de estrógeno y progesterona de las hembras de Leptodactylus macrosternum y su relación con la temperatura y la precipitación. Los especímenes fueron recolectados en el área Horto Florestal Olho d'Água da Bica - HFOB (06 ° 49´20''S / 36 ° 15´85''W) en el municipio de Cuité, semiárido brasileño, cada 15 días, ocurrieron entre los meses de mayo a agosto y de noviembre a diciembre. Se utilizó la cuantificación de la densidad poblacional de los tipos de ovocitos y las dosis de progesterona y niveles de estrógenos para determinar la actividad reproductiva de la espécie. Las densidades de población de ovogonias y ovocitos II no variaron significativamente durante el experimento. La densidad poblacional de ovocito I fue significativamente mayor durante los meses de junio (p = 0.04) y agosto (p = 0.03). Los valores más altos para la densidad poblacional de ovocito III y IV se encontraron en los meses de mayo (p = 0.04) y junio (p = 0,04). Las cantidades de concentraciones de estrógeno (pg / ml) y progesterona (pg / ml) no cambiaron significativamente durante los períodos de muestreo. La densidad de población de los tipos de ovocitos III y IV mostró dependencia de la variación de las precipitaciones. Em regiões tropicais com forte sazonalidade, a atividade reprodutiva de muitas espécies ocorre nos períodos mais adequados do ano. Portanto, vários ajustes fisiológicos e comportamentais são necessários durante seu ciclo reprodutivo. Este estudo tem como objetivo avaliar possíveis alterações nos parâmetros histológicos e morfométricos dos ovários, bem como nos níveis plasmáticos de estrogênio e progesterona de fêmeas de Leptodactylus macrosternum e sua relação com a temperatura e precipitação. Os espécimes foram coletados na área do Horto Florestal Olho d'Água da Bica - HFOB (06 ° 49´20''S / 36 ° 15´85''W) no município de Cuité, semiárido brasileiro, a cada 15 dias, apenas entre os meses de maio a agosto e de novembro a dezembro. A quantificação da densidade populacional dos tipos de ovócitos e dosagens de progesterona e níveis de estrogênio foram utilizados para determinar a atividade reprodutiva das espécies. As densidades populacionais de oogônios e oócitos II não variaram significativamente durante os meses em que os animais foram amostrados. As densidades populacionais do oócito I foram significativamente maiores durante os meses de junho (p = 0,04) e agosto (p = 0,03). Os maiores valores para a densidade populacional do oócito III e IV foram encontrados nos meses de maio (p = 0,04 ) e junho (p = 0,04). As quantidades de concentrações de estrogênio (pg / ml) e progesterona (pg / ml) não se alteraram significativamente ao longo dos períodos de amostragem. A densidade populacional dos tipos de ovócitos III e IV mostrou dependência da variação da precipitação.

Glucosinolates are token stimuli in host selection of many crucifer specialist insects, but the underlying molecular basis for host selection in these insects remains enigmatic. Using a combination of behavioral, electrophysiological, and molecular methods, we investigate glucosinolate receptors in the cabbage butterfly Pieris rapae. Sinigrin, as a potent feeding stimulant, elicited activity in larval maxillary lateral sensilla styloconica, as well as in adult medial tarsal sensilla. Two P. rapae gustatory receptor genes PrapGr28 and PrapGr15 were identified with high expression in female tarsi, and the subsequent functional analyses showed that Xenopus oocytes only expressing PrapGr28 had specific responses to sinigrin; when ectopically expressed in Drosophila sugar sensing neurons, PrapGr28 conferred sinigrin sensitivity to these neurons. RNA interference experiments further showed that knockdown of PrapGr28 reduced the sensitivity of adult medial tarsal sensilla to sinigrin. Taken together, we conclude that PrapGr28 is a gustatory receptor tuned to sinigrin in P. rapae, which paves the way for revealing the molecular basis of the relationships between crucifer plants and their specialist insects., Author summary Preference of crucifer specialist insects to glucosinolates is well known in the field of insect-plant interactions, but its molecular basis is unclear. This study uses an integrative approach to investigate the molecular basis of glucosinolate detection by gustatory receptor neurons in the larval mouthparts and adult forelegs of the cabbage butterfly Pieris rapae, and finally reveal that PrapGr28 is a bitter receptor tuned to sinigrin. The current work takes a significant step towards identifying gustatory receptors tuned to glucosinolates, crucial recognition signals in crucifer host plants, providing insights into co-evolution of herbivorous insects and their host plants.

Boron (B) is essential for vascular plants. Rapeseed (Brassica napus) is the second leading crop source for vegetable oil worldwide, but its production is critically dependent on B supplies. BnaA3.NIP5;1 was identified as a B-efficient candidate gene in B. napus in our previous QTL fine mapping. However, the molecular mechanism through which this gene improves low-B tolerance remains elusive. Here, we report genetic variation in BnaA3.NIP5;1 gene, which encodes a boric acid channel, is a key determinant of low-B tolerance in B. napus. Transgenic lines with increased BnaA3.NIP5;1 expression exhibited improved low-B tolerance in both the seedling and maturity stages. BnaA3.NIP5;1 is preferentially polar-localized in the distal plasma membrane of lateral root cap (LRC) cells and transports B into the root tips to promote root growth under B-deficiency conditions. Further analysis revealed that a CTTTC tandem repeat in the 5’UTR of BnaA3.NIP5;1 altered the expression level of the gene, which is tightly associated with plant growth and seed yield. Field tests with natural populations and near-isogenic lines (NILs) confirmed that the varieties carried BnaA3.NIP5;1Q allele significantly improved seed yield. Taken together, our results provide novel insights into the low-B tolerance of B. napus, and the elite allele of BnaA3.NIP5;1 could serve as a direct target for breeding low-B-tolerant cultivars., Author summary Boron (B) deficiency severely rapeseed (Brassica napus) yields in most high rainfall areas worldwide, and genetic improvement is an effective strategy for addressing the problem. Here we show that BnaA3.NIP5;1, encoding a boric acid channel, is a key determinant of the low-B tolerance in B. napus. Our results demonstrate that BnaA3.NIP5;1 is preferentially located in the distal side plasma membrane of lateral root cap (LRC) cells and transports B into meristem zone to promote root growth under B limitation, which provide insights into the LRC’s function in mineral nutrition. A CTTTC tandem repeat in the 5’UTR of BnaA3.NIP5;1 altered the expression level of the gene, which is tightly associated with plant growth and seed yield under low-B conditions. The functional gene and elite allele could be useful in rapeseed breeding.

The formation and maintenance of microtubules requires their polymerisation, but little is known about how this polymerisation is regulated in cells. Focussing on the essential microtubule bundles in axons of Drosophila and Xenopus neurons, we show that the plus-end scaffold Eb1, the polymerase XMAP215/Msps and the lattice-binder Tau co-operate interdependently to promote microtubule polymerisation and bundle organisation during axon development and maintenance. Eb1 and XMAP215/Msps promote each other’s localisation at polymerising microtubule plus-ends. Tau outcompetes Eb1-binding along microtubule lattices, thus preventing depletion of Eb1 tip pools. The three factors genetically interact and show shared mutant phenotypes: reductions in axon growth, comet sizes, comet numbers and comet velocities, as well as prominent deterioration of parallel microtubule bundles into disorganised curled conformations. This microtubule curling is caused by Eb1 plus-end depletion which impairs spectraplakin-mediated guidance of extending microtubules into parallel bundles. Our demonstration that Eb1, XMAP215/Msps and Tau co-operate during the regulation of microtubule polymerisation and bundle organisation, offers new conceptual explanations for developmental and degenerative axon pathologies., Author summary Axons are the up-to-meter-long processes of nerve cells that form the cables wiring our nervous system. Once established, they must survive for a century in humans. Improper extension of axons leads to neurodevelopmental defects, and age- or disease-related neurodegeneration usually starts in axons. Axonal architecture and function depend on bundles of filamentous polymers, called microtubules. These bundles run all along the axonal core, and their disruption correlates with axon decay. How these axonal microtubule bundles are formed and dynamically maintained is little understood. We bridge this knowledge gap by studying how different classes of microtubule-binding proteins may regulate these processes. Here we show how three proteins of very different function, Eb1, XMAP215 and Tau, cooperate intricately to promote the polymerisation processes that form new microtubules during axon development and maintenance. If either protein is dysfunctional, polymerisation is slowed down and newly forming microtubules fail to align into proper bundles. These findings provide new explanations for the decay of microtubule bundles, hence axons. To unravel these mechanisms, we used the fruit fly as a powerful organism for biomedical discoveries. We then showed that the same mechanisms act in frog axons, suggesting they might apply also to humans.

We report the presence of Alsodes verrucosus (Philippi, 1902) in Cayutué, Los Lagos Region, Chile, extending this species’ geographic distribution 65 km south of the previously southernmost record. An updated distribution map is provided for A. verrucosus and previous records in the literature are discussed.

The Lesser Sundas Archipelago is comprised of two parallel chains of islands that extend between the Asian continental shelf (Sundaland) and Australo-Papuan continental shelf (Sahul). These islands have served as stepping stones for taxa dispersing between the Asian and Australo-Papuan biogeographical realms. While the oceanic barriers have prevented many species from colonizing the archipelago, a number of terrestrial vertebrate species have colonized the islands either by rafting/swimming or by human introduction. Here, we examine phylogeographic structure within the Lesser Sundas for three snake, two lizard and two frog species that each has a Sunda Shelf origin. These species are suspected to have recently colonized the archipelago, though all have inhabited the Lesser Sundas for over 100years. We sequenced mtDNA from 231 samples to test whether there is sufficiently deep genetic structure within any of these taxa to reject human-mediated introduction. Additionally, we tested for genetic signatures of population expansion consistent with recent introduction and estimated the ages of Lesser Sundas clades, if any exist. Our results show little to no genetic structure between populations on different islands in five species and moderate structure in two species. Nucleotide diversity is low for all species, and the ages of the most recent common ancestor for species with monophyletic Lesser Sundas lineages date to the Holocene or late Pleistocene. These results support the hypothesis that these species entered the archipelago relatively recently and either naturally colonized or were introduced by humans to most of the largerislands in the archipelago within a short time span.

Multiple pathogens coexist in nature, and hence, host species often encounter several pathogens simultaneously. The sequence in which the host encounters the parasites influences interactions between parasites and host pathology. Here, the effects of infection by two cercaria (larvae of trematodes) types, pleurolophocercous cercaria of Acanthostomum burminis and a furcocercous cercaria, on the tadpoles of common hourglass tree frog (Polypedates cruciger) were examined. Ten days posthatch, tadpoles (Gosner stage 27/28) were used for infection exposures. First, in a single infection each cercaria type was introduced to the tadpoles separately. Second, coinfection of the two cercaria was carried out by alternating the sequences of exposure. For all the experiments, appropriate controls were instituted. Tadpoles of all groups exposed to parasites had lower survival levels compared to controls. Among the four groups exposed, the highest survival was observed in the coinfection when furcocercous was introduced first (82.5%). The lowest survival was observed in the coinfection when the A. burminis cercaria was introduced first (65.0%). In the coinfections, when A. burminis was introduced prior to furcocercous, survival of the tadpoles was reduced by 17.0% compared to the exposures of furcocercous prior to A. burminis. Prior infection with A. burminis induced negative effect on the host with an increased infection severity, while prior infection with furcocercous had reduced infection severity than lone exposures. These results suggest that furcocercous infections can be beneficial for hosts challenged with A. burminis provided that A. burminis exposure occurs second. None of the treatments had an effect on the growth of the tadpoles, but lengthening of developmental period was observed in some exposures. All exposed tadpoles developed malformations which were exclusively axial—kyphosis and scoliosis. However, there was no difference in the number of malformed individuals in the single infection (19.0%–25.0%) compared to coinfection (20.0%–22.5%) or between coinfections. The results suggest that the sequence of parasite exposure affects host–parasite interactions and hence the disease outcomes. Understanding the effects of coinfection on disease outcomes for hosts provides insight into disease dynamics.

We studied the species of amphibians from Serranía de Las Quinchas, in the mid Magdalena Valley region of Colombia, comparing the data from a recent fieldwork with museum records and literature. We present a list of 50 species of amphibians (Anura and Caudata). In our survey, we recorded 36 species, of which 6 represented new records. One of the most interesting record is that of the genus Oedipina, since this is the first time the genus is recorded for the Cordillera Oriental. The number of species is what would be expected in a humid tropical forest of the mid-Magdalena river valley, given the co-occurrence of amphibian faunas distributed in the sub-Andean, Caribbean, and Chocó biogeographic regions.

The spatial distribution of species has long sparked interest among ecologists and biogeographers, increasingly so in studies of species responses to climate change. However, field studies on spatial patterns of distribution, useful to inform conservation actions at local scales, are still lacking for many regions, especially the tropics. We studied elevational trends and species‐area relationships among anurans in wetland habitats within Volcanoes National Park (VNP) in Rwanda, part of the biodiverse Albertine Rift region. In VNP, wetlands are key sites for anuran reproduction, and anurans are likely threatened by wetland desiccation which has occurred for the last few decades. Between 2012 and 2017, we sampled anuran communities in ten VNP wetlands located along an elevational gradient of c. 600 m (from 2,546 to 3,188 m a.s.l.) and found at least eight species, including at least two Albertine Rift Endemics. We show that species richness, diversity, and abundance likely decline with a decrease in wetland size and with an increase in elevation, though additional sampling (e.g., at night) might be needed to derive definite conclusions. Larger wetlands at lower elevations contained most species and individuals, which indicates the potential threat of wetland size reduction (through desiccation) for anuran conservation. However, we also found that wetlands differed in species composition and that some species (e.g., Sclerophrys kisoloensis) were likely restricted in distribution to only a few of the smaller wetlands—suggesting that the conservation of each individual wetland should be prioritized, regardless of size. We propose that all wetlands in VNP require additional conservation measures, which should be based on knowledge gathered through long‐term monitoring of anuran communities and research on drivers of wetland decline. Only such extended research will allow us to understand the response of anurans in VNP to threats such as climate change and wetland desiccation.

Composición de la dieta de la rana de Kárpatos (Pelophylax cerigensis): ¿qué come la rana más amenazada de Europa? La rana de Kárpatos (Pelophylax cerigensis) es considerada la rana más amenazada de Europa. Aquí evaluamos su ecología alimentaria y examinamos 76 individuos de las dos poblaciones conocidas usando el método del lavado de estómago. También medimos el peso corporal, la longitud desde el hocico hasta la cloaca y el ancho de la boca de las ranas y el ancho y largo de las presas. La dieta de Pelophylax cerigensis, compuesta principalmente por Coleoptera, Aranean, Isopoda e Hymenoptera, es similar a la de otras especies de ranas verdes de las zonas adyacentes. Las dos poblaciones difieren en el tamaño corporal, pero presentan valores similares de abundancia y frecuencia de presas. Parece que P. cerigensis sigue una estricta estrategia de alimentación. El estudio de la disponibilidad de presas en sus hábitats aportará información valiosa., The Karpathos marsh frog (Pelophylax cerigensis) is considered the most endangered frog in Europe. Here we assess its feeding ecology and examine 76 individuals from the two known populations using the stomach flushing method. We also measured body weight, snout–vent length, mouth width and prey width and length. Pelophylax cerigensis follows the feeding pattern of green frogs of the adjacent areas, with Coleoptera, Araneae, Isopoda and Hymenoptera being the main prey groups. The two populations differed in body size but had similar values of prey abundance and frequency. It seems that P. cerigensis follows a strict feeding strategy. Further research on prey availability in its habitats will provide valuable insight.

International audience; : Hydrological models have become practical tools to understand impacts in water resources and to support the development of management policies. Precipitation is a major driving force of hydrological processes and is one of the main input datasets for hydrological models. However, gauge measurements have several issues, such as incomplete areal coverage and deficiencies over most tropical regions. The precipitation data obtained by remote sensing is an alternative in areas where data is scarce or not available, like in the tropics. However, the feasibility of using satellite-based precipitation products for simulating streamflow needs to be verified, for different hydrological models and basins. Congo River Basin is one of the least studied major river basins in the world and suffers from the scarcity and difficulty in accessing rain gauge data, which makes satellite precipitation estimates necessary for hydrological studies. In this study, we analyzed twenty-three satellite-based precipitation products, acquired from Frequent Rainfall Observations on GridS (FROGS) daily precipitation database. We evaluated these precipitation products over the Congo River Basin using the hydrological model SWAT (Solid & Water Assessment Tool) for streamflow and water balance components at basin scale. Our findings showed that the products based on satellite-only source tend to overestimate the rainy season peaks in comparison with the 3B42_V7 product. On the other hand, the satellite products that consider gauge calibration presented better agreements between each other. The hydrological model was able to reproduce the general precipitation products characteristics, while the gauge-adjusted satellite products performed better than those without gauge adjustments. Thus, the overall precipitation patterns have a crucial effect on model’s performance and leads to different streamflow and water balance components values. The choice of rainfall product has a significant importance in the interpretation of the simulated hydrological cycle.

Intracellular parasites of the phylum Apicomplexa are dependent on the scavenging of essential amino acids from their hosts. We previously identified a large family of apicomplexan-specific plasma membrane-localized amino acid transporters, the ApiATs, and showed that the Toxoplasma gondii transporter TgApiAT1 functions in the selective uptake of arginine. TgApiAT1 is essential for parasite virulence, but dispensable for parasite growth in medium containing high concentrations of arginine, indicating the presence of at least one other arginine transporter. Here we identify TgApiAT6-1 as the second arginine transporter. Using a combination of parasite assays and heterologous characterisation of TgApiAT6-1 in Xenopus laevis oocytes, we demonstrate that TgApiAT6-1 is a general cationic amino acid transporter that mediates both the high-affinity uptake of lysine and the low-affinity uptake of arginine. TgApiAT6-1 is the primary lysine transporter in the disease-causing tachyzoite stage of T. gondii and is essential for parasite proliferation. We demonstrate that the uptake of cationic amino acids by TgApiAT6-1 is ‘trans-stimulated’ by cationic and neutral amino acids and is likely promoted by an inwardly negative membrane potential. These findings demonstrate that T. gondii has evolved overlapping transport mechanisms for the uptake of essential cationic amino acids, and we draw together our findings into a comprehensive model that highlights the finely-tuned, regulated processes that mediate cationic amino acid scavenging by these intracellular parasites., Author summary The causative agent of toxoplasmosis, Toxoplasma gondii, is a versatile intracellular parasite that can proliferate within nucleated cells of warm-blooded organisms. In order to survive, T. gondii parasites must scavenge the cationic amino acids lysine and arginine from their hosts. In a previous study, we demonstrated that a plasma membrane-localized protein called TgApiAT1 facilitates the uptake of arginine into the parasite. We found that parasites lacking TgApiAT1 could proliferate when cultured in medium containing high concentrations of arginine, suggesting the existence of an additional uptake pathway for arginine. In the present study, we demonstrate that this second uptake pathway is mediated by TgApiAT6-1, a protein belonging to the same solute transporter family as TgApiAT1. We show that TgApiAT6-1 is the major lysine transporter of the parasite, and that it is critical for parasite proliferation. Furthermore, we demonstrate that TgApiAT6-1 can transport arginine into parasites under conditions in which arginine concentrations are high and lysine concentrations are comparatively lower. These data support a model for the finely-tuned acquisition of essential cationic amino acids that involves multiple transporters, and which likely contributes to these parasites being able to survive and proliferate within a wide variety of host cell types.

Onchocerciasis (river blindness), caused by the filarial worm Onchocerca volvulus, is a neglected tropical disease mostly affecting sub-Saharan Africa and is responsible for >1.3 million years lived with disability. Current control relies almost entirely on ivermectin, which suppresses symptoms caused by the first-stage larvae (microfilariae) but does not kill the long-lived adults. Here, we evaluated emodepside, a semi-synthetic cyclooctadepsipeptide registered for deworming applications in companion animals, for activity against adult filariae (i.e., as a macrofilaricide). We demonstrate the equivalence of emodepside activity on SLO-1 potassium channels in Onchocerca volvulus and Onchocerca ochengi, its sister species from cattle. Evaluation of emodepside in cattle as single or 7-day treatments at two doses (0.15 and 0.75 mg/kg) revealed rapid activity against microfilariae, prolonged suppression of female worm fecundity, and macrofilaricidal effects by 18 months post treatment. The drug was well tolerated, causing only transiently increased blood glucose. Female adult worms were mostly paralyzed; however, some retained metabolic activity even in the multiple high-dose group. These data support ongoing clinical development of emodepside to treat river blindness., Author summary Onchocerciasis (river blindness), caused by the parasitic worm Onchocerca volvulus, is a devastating neglected tropical disease affecting sub-Saharan Africa with an overall impact of >1.3 million years lived with disability. Current control relies mainly on a single drug, ivermectin, which suppresses symptoms caused by the first-stage larvae (microfilariae) but does not kill the long-lived adults. The identification of a drug that can safely eliminate adult worms (i.e., a macrofilaricide) is a major research objective for onchocerciasis. We evaluated the anthelminthic activity of emodepside, a veterinary wormer, in cattle infected with a close relative of O. volvulus (Onchocerca ochengi) before conducting pharmacokinetic modelling to estimate drug distribution in humans. Emodepside as single or 7-day treatments at two doses produced rapid activity against O. ochengi microfilariae, prolonged suppression of female worm fecundity, and paralysed most female worms by 18 months, although some remained metabolically active even in the multiple high-dose treatment group. The drug was well tolerated, causing only transiently increased blood glucose. Thus, emodepside shows slow but significant efficacy against adult O. ochengi in naturally infected cattle, meeting the criteria for a safe macrofilaricidal drug. Our data support the ongoing clinical development of emodepside for the treatment of human onchocerciasis.

River regulation or flow modification—the hydrological alteration of flow by dams and diversions—has been implicated as a cause of fundamental change to downstream aquatic ecosystems. Flow modification changes the patterns and functionality of the natural flow regime and has the potential to restrict population connectivity and gene flow in river‐dependent organisms. Since population connectivity and the maintenance of genetic diversity are fundamental drivers of long‐term persistence, understanding the extent flow modification impacts these critical attributes of genetic health is an important goal for long‐term conservation. Foothill yellow‐legged frogs (Rana boylii) were historically abundant throughout many western rivers but have declined since the onset of flow regulation. However, the extent to which R. boylii populations in rivers with altered flow regimes are maintaining connectivity and genetic diversity is unknown. Here, we use population genomics to investigate the impacts of flow alteration on R. boylii to explore their potential for long‐term persistence under continued flow modification. We found R. boylii in rivers with flow modification showed striking patterns of isolation and trajectories of genetic diversity loss relative to unregulated rivers. For example, flow modification explained the greatest amount of variance in population genetic differentiation compared with other covariates including geographic distance. Importantly, patterns of connectivity and genetic diversity loss were observed regardless of flow alteration level but were most prominent in locations with the greatest flow modification intensity. Although our results do not bode well for long‐term persistence of R. boylii populations under current flow regulation regimes, they do highlight the power of genetic monitoring for assessing population health in aquatic organisms.