Cutaneous Bacteria of Confiscated Telmatobius culeus in Lima, Peru.

The Lake Titicaca frog is endangered due to threats such as water pollution, introduced species, and overharversting for markets, where people consume them as frog juice. This study, conducted June to November 2012, aimed to determinate the bacteria microflora living on the skin of frogs confiscated from the La Parada market, Lima, Peru, and housed individually in the Laboratory of Wildlife at the Faculty of Veterinary Medicine and Zootechnic of the Universidad Peruana Cayetano Heredia in Lima, Peru. Samples collected with sterile swabs and cultured on blood, tryptic soy, and MacConkey agars were investigated using commercially available test kits, to investigate the commonly encountered bacterial and potentially zoonotic microorganisms associated with their consumption. We found three species of zoonotic concern in the genus Vibro: Vibrio alginolyticus, Vibro cholerae, and Vibro fluvialis. Other Gram-negative species cultured included two different colonies of Aeromonas hydrophila, or Aeromonas caviae or Aeromonas sobria; Pseudomona luteola; one example of Weeksella virosa or Empedobacter brevis; and Citrobacter freundii. Gram-positive bacteria detected were Staphylococcus spp., Micrococcus spp., and Erysipelothrix rhusiopathiae. We recommend against the consumption of this frog due to the pathogens it may carry that could cause serious illness among consumers and in vendors who handle animals.

Rediscovery of the enigmatic Andean frog Telmatobius halli Noble (Anura: Telmatobiidae), re-description of the tadpole and comments on new adult's characters, type locality and conservation status.

We report the rediscovery of Telmatobius halli (Hall's water frog), which had not been found since its description (over 80 years) since its type locality was not clearly established. "Aguas Calientes" near Ollagüe is hypothesized as the original type locality where Frank Gregory Hall collected the type material in 1935. The tadpole is re-described, and new data on the external and internal morphology of adults is provided. These new morphological data are compared with Telmatobius spp. inhabiting geographically close to T. halli in Chile and Bolivia. In addition, comments on its ecology, conservation, and taxonomic status in relation with other Telmatobius spp. inhabiting nearby areas in Ascotán and Carcote salt pans are provided. No evidence of Batrachochytrium dendrobatidis and Ranavirus infection was found in T. halli and a sympatric amphibian species. Our work supports the validity of T. halli and suggests this species should be considered as Data Deficient in the IUCN Red List assessment until taxonomic issues are resolved.

Aspectos ecológicos de Telmatobius atacamensis (Anura: Telmatobiidae), un microendemismo de la Puna, Salta-Argentina

Resumen Telmatobius atacamensis es un anuro microendémico amenazado de la Puna salteña. En el presente estudio se identificaron los núcleos de reproducción caracterizando sus poblaciones. Para ello se realizó un muestreo anual en los ríos Los Patos y San Antonio, estableciéndose un total de diez sitios, registrándose pH, temperatura y conductividad eléctrica en los cuerpos de agua. Se empleó la técnica de encuentro visual (TEV) para la detección de los individuos. Se establecieron tres categorías de desarrollo larval, prometamórfico (G1), premetamórfico (G2) y clímax metamórfico (G3). Se identificaron seis núcleos reproductivos, cuatro en río San Antonio y dos en río Los Patos. Los individuos se localizaron en refugios subacuáticos con un rango reducido de conductividad eléctrica y uno amplio de temperatura mientras que el pH fue alcalino. Ambos ríos mostraron diferencias en cuanto a la frecuencia relativa de categorías de estadio larval. Las abundancias de esta etapa fue significativamente diferente entre ambos (ANOVA F= 109.41 p<0.000) siendo mayor para el río Los Patos. Los individuos adultos fueron registrados en ambos ríos durante todo el ciclo, con diferencias significativas en sus abundancias (T= 19.83 p<0.0001), resultando mayor para el río Los Patos. El presente trabajo aportaría información relevante para la evaluación del estado actual de conservación y la elaboración objetiva de estrategias para el diseño de monitores efectivos, comparaciones poblacionales futuras y protección de la especie.

Abstract Telmatobius atacamensis is a microendemic threatened anuran of the Puna of Salta, Argentina. In this study, we identify reproduction nuclei and we characterize the anurans populations. To do this, an annual sampling was made in Los Patos and San Antonio rivers and we selected ten sampling sites. We measured pH, temperature and electrical conductivity at each site. The visual encounter survey (VES) was used in order to identify the individuals. Three categories of larval development were established: prometamorphic (G1), premetamorphic (G2) and metamorphic climax (G3). We found a total of six reproductive nuclei, four in San Antonio River and two in Los Patos River. The individuals were located in underwater shelters with a low range of electrical conductivity and in a wide range of temperature while the pH was alkaline. The relative abundance at the larval stages were significantly different between the rivers (ANOVA F= 109.41 p<0.000), the highest abundance was detected in Los Patos River. The adults were sampled in both rivers during the year with significant differences in their abundance (T= 19.83 p<0.0001), being the highest in Los Patos River. This study would contribute significant information to the assessment of the current conservation status and the objective development of strategies for the design of effective monitoring, further population comparisons and species protection.

Evaluación de la abundancia relativa de Telmatobius culeus en la zona litoral del lago Titicaca, Perú / Assessment of relative abundance of Telmatobius culeus in littoral zone of Lake Titicaca, Peru

El estudio se realizó en 13 zonas del litoral peruano del lago Titicaca, durante la época seca, desde junio hasta agosto de 2017. En cada zona se determinó la abundancia relativa y uso de microhábitat de Telmatobius culeus. En las zonas de evaluación se utilizó buceo con esnórquel para realizar en cada una tres transectos de 100 m x 2 m, con un total de 38 transectos evaluados. Se registraron 45 individuos en solo seis zonas de evaluación. El mayor número de individuos se encontraron en la zona de Perka Norte, con una abundancia relativa de 10.33 individuos/hora/02 personas a una profundidad de 1.5 m; en siete zonas no se registró ningún individuo. Los individuos fueron encontrados en los siguientes micro hábitats: 17 fueron encontrados sobre la vegetación acuática, 11 en arena combinado con vegetación acuática, 9 en roca combinado con vegetación acuática, 5 en fango combinado con vegetación acuática, 2 en roca y 1 en arena. a la vez se realizó análisis de diferentes parámetros de calidad de agua en cada zona de estudio, como: temperatura, pH, alcalinidad, dureza y oxígeno.

The Telmatobius culeus relative abundance and its microhabitat use was determined. In 13 areas of Peruvian littoral of Lake Titicaca was carried out, during the dry season, since June to August 2017. In each the evaluation areas, snorkeling was used to perform three 100 x 2 m transects. A total of 38 transects were evaluated. 45 individuals were registered in only six evaluation zones. The largest number of individuals were found in the North Perka area, with a relative abundance of 10.33 individuals/hour/02 people, at a depth of 1.5 m; in seven zones no individual was registered. Individuals were found in the following micro habitats: 17 were found on aquatic vegetation, 11 in sand combined with aquatic vegetation, 9 in rock combined with aquatic vegetation, 5 in mud combined with aquatic vegetation, 2 in rock and 1 in sand. At the same time, analyzes of different water quality parameters were performed in each study area, such as: temperature, pH, alkalinity, hardness and oxygen.

Widespread occurrence of the antifungal cutaneous bacterium Janthinobacterium lividum on Andean water frogs threatened by fungal disease.

Amphibian diversity has declined due to the infectious disease chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd). Coexistence between amphibian hosts and this pathogen in some locations is attributed to the presence of the cutaneous bacterium Janthinobacterium lividum (Jliv). This microbe inhibits the growth of Bd on the host, reduces morbidity, and improves survival. Andean water frogs in the genus Telmatobius seem to be particularly vulnerable to the disease yet populations of T. intermedius and T. marmoratus persist in southern and central Peru. We investigated the presence of Jliv on these 2 frog species and assessed the relationship of Jliv presence with prevalence and intensity of Bd infection. By sampling 125 frogs from 7 streams (3323-3950 m a.s.l.) and 27 from a city market, we found spatial variation in the mutualism among populations (range 0-40% proportion of Jliv-positives). Overall, 57% of frogs were infected with Bd, 12.5% of frogs hosted both Jliv and Bd, while 7.2% hosted just Jliv. We found that the probability of an individual being infected with Bd was independent of the presence of Jliv; however, we did detect a protective effect of Jliv with respect to intensity of infection. The extent of Jliv distribution in the high Andes stands in stark contrast to the rarity of Jliv on frogs in lower elevation cloud forest biomes.

Redescription of Hedruris mucronifer (Nematoda: Hedruridae) parasite of Telmatobius schreiteri (Anura, Telmatobiidae) from Tucumán, Argentina.

The species Hedruris mucronifer Schuurmans Stekhoven, 1952 parasite of Telmatobius schreiteri from Tafí del Valle, Tucumán, Argentina is herein redescribed. The main morphological characters used for its identification are illustrated. The lectotype and paralectotype are designated, respectively. The specific epithet of the host is rectified.

Development and characterization of 22 polymorphic microsatellites of the Andean frog Telmatobius chusmisensis (Anura, Telmatobius) and cross amplification in seven Chilean species of the genus.

The genus Telmatobius Wiegmann, 1834 is composed of a wide variety of species and is one of the most diverse native components of the high-altitude Andean environments. The species of the genus present in Chile are considered as endangered, critically endangered or data deficient. We isolated and evaluated 44 microsatellites in 80 individuals of 8 species of Telmatobius present in Chile, obtaining 22 polymorphic microsatellite loci for Telmatobius chusmisensis. The cross-amplification test was successful in all other species tested. For the first time, microsatellite markers are described for Telmatobius. The description of these primers will be useful for further genetic studies for T. chusmisensis and other species of the same genus; allowing further analyses of population structuring, dispersal patterns, recent demographic history and population effective size. This information is also significant to undertake conservation actions for the species of the genus Telmatobius, since most species have conservation issues.

The taxonomic status of two Telmatobius frog species (Anura: Telmatobiidae) from the western Andean slopes of northernmost Chile.

On the basis of molecular and morphological evidence, we evaluated the taxonomic identity of two species of Andean frogs of the genus Telmatobius: Telmatobius pefauri and T. zapahuirensis, present in the western Andean slopes at the northern extreme of Chile. We also investigated the taxonomic assignment of five populations of Telmatobius recently discovered around the type localities of these two species. The results indicate that T. pefauri inhabits, not only Murmuntani its type locality, but also the montane localities of Belén, Copaquilla, Lupica, Saxamar and Socoroma. Our study also shows that T. pefauri and T. zapahuirensis are the same taxon. Therefore, Telmatobius zapahuirensis Veloso, Sallaberry, Navarro, Iturra, Valencia, Penna & Díaz, 1982 would be a subjective junior synonym of Telmatobius pefauri Veloso & Trueb, 1976.

Long-term monitoring of tropical alpine habitat change, Andean anurans, and chytrid fungus in the Cordillera Vilcanota, Peru: Results from a decade of study.

The Cordillera Vilcanota in southern Peru is the second largest glacierized range in the tropics and home to one of the largest high-alpine lakes, Sibinacocha (4,860 m). Here, Telmatobius marmoratus (marbled water frog), Rhinella spinulosa (Andean toad), and Pleurodema marmoratum (marbled four-eyed frog) have expanded their range vertically within the past century to inhabit newly formed ponds created by ongoing deglaciation. These anuran populations, geographically among the highest (5,200-5,400 m) recorded globally, are being impacted by the chytrid fungus Batrachochytrium dendrobatidis (Bd), and the disease it causes, chytridiomycosis. In this study, we report results from over a decade of monitoring these three anuran species, their habitat, and Bd infection status. Our observations reveal dynamic changes in habitat including ongoing rapid deglaciation (18.4 m/year widening of a corridor between retreating glaciers from 2005 to 2015), new pond formation, changes in vegetation in amphibian habitat, and widespread occurrence of Bd in amphibians in seven sites. Three of these sites have tested positive for Bd over a 9- to 12-year period. In addition, we observed a widespread reduction in T. marmoratus encounters in the Vilcanota in 2008, 2009, and 2012, while encounters increased in 2013 and 2015. Despite the rapid and dynamic changes in habitat under a warming climate, continued presence of Bd in the environment for over a decade, and a reduction in one of three anuran species, we document that these anurans continue to breed and survive in this high Andean environment. High variability in anuran encounters across sites and plasticity in these populations across habitats, sites, and years are all factors that could favor repopulation postdecline. Preserving the connectivity of wetlands in the Cordillera Vilcanota is therefore essential in ensuring that anurans continue to breed and adapt as climate change continues to reshape the environment.

Batrachochytrium dendrobatidis in Confiscated Telmatobius in Lima, Peru.

The Peruvian Andes are the home of 27 species of frogs of the genus Telmatobius, many of which are critically endangered. Illegal trade of adult frogs for purported medical properties likely represents the major threat facing these species. This activity, besides reducing their populations, may contribute to the dissemination of the fungus Batrachochytrium dendrobatidis (Bd), which causes chytridiomycosis, a disease posing a threat to many populations of amphibians. We screened frogs confiscated by the Administration of Forestry and Wildlife in Lima, Peru, for Bd. We used real-time PCR to diagnose Bd at the Laboratory of Wildlife, Faculty of Veterinary Medicine and Zootecnics, Universidad Peruana Cayetano Heredia, in Lima and Pisces Molecular Laboratory in Boulder, Colorado, US. Of 62 samples collected during this study, 60% (37) were PCR positive for Bd, confirming that illegal trade of amphibians can pose a risk for disseminating Bd.

Independent Evolution of Suction Feeding in Neobatrachia: Feeding Mechanisms in Two Species of Telmatobius (Anura:Telmatobiidae).

The most common feeding mechanism among aquatic vertebrates as fishes, turtles, and salamanders is inertial suction. However, among the more than 6,400 species of anurans, suction feeding occurs only in pipids. Pipidae is a small basal lineage relative to Neobatrachia, an enormous clade that contains about 96% of extant anurans. The Andean neobatrachian frogs of the genus Telmatobius include strictly aquatic and semiaquatic species. Diet analyses indicate that some species of Telmatobius feed on strictly aquatic prey, but until now their feeding mechanisms have been unknown. Herein, the feeding mechanisms in two species of Telmatobius, that represent the two predominant modes of life in the genus, are explored. The semiaquatic T. oxycephalus and the fully aquatic T. rubigo are studied using high-speed cinematography and standard anatomical techniques to provide a qualitative approach to feeding behavior and a detailed morphological description of the mouth, tongue, hyoid and related muscles. T. oxycephalus uses similar mechanisms of aquatic prey capture as do the vast majority of anurans that are capable of forage in water, whereas the fully aquatic T. rubigo is an inertial suction feeder. This is the first report of an objective record of this unique feeding behavior in a Neobatrachian. Several morphological characters seem to be related with this function and are convergent with those of pipids.

The complete mitochondrial genome of the Chilean endemic frog Telmatobius chusmisensis Formas, Cuevas & Nuñez, 2006 (Anura, Telmatobiidae).

Telmatobius chusmisensis is an endemic frog of northern Chile that is only known at its type locality, Chusmisa. In this study, the complete mitochondrial genome of Telmatobius chusmisensis was assembled using high-throughput sequencing data, yielding a circular genome of 19 312 bp with a nucleotide composition of A = 30.8%, C = 24.4%, G = 13.6% and T = 31.2%. Its gene composition and structure were similar to other anuran genomes available: 13 protein-coding genes, two rRNA genes, 22 tRNA genes and the D-loop region. Phylogenetic analysis using Bayesian inference (BI) and maximum likelihood (ML) showed that Telmatobius chusmisensis, T. bolivianus and T. vellardi are a highly supported monophyletic group. This genome information will allow us to gain a better understanding into phylogenetic and phylogeographic relationships in the genus Telmatobius.

Evolution and Conservation on Top of the World: Phylogeography of the Marbled Water Frog (Telmatobius marmoratus Species Complex; Anura, Telmatobiidae) in Protected Areas of Chile.

The Andean Altiplano has served as a complex setting throughout its history, driving dynamic processes of diversification in several taxa. We investigated phylogeographic processes in the Telmatobius marmoratus species complex occurring in this region by studying the geographic patterns of genetic variability, genealogies, and historical migration, using the cytochrome b (cyt-b) gene as a marker. DNA sequences from Telmatobius gigas and Telmatobius culeus, Bolivian species with an uncertain taxonomic status, were also included. Additionally, we evaluated the phylogenetic diversity (PD) represented within Chilean protected areas and the complementary contribution from unprotected populations. Phylogenetic reconstructions from 148 cyt-b sequences revealed 4 main clades, one of which corresponded to T. culeus. T. gigas was part of T. marmoratus clade indicating paraphyletic relationships. Haplotypes from Chilean and Bolivian sites were not reciprocally monophyletic. Geographic distribution of lineages, spatial Bayesian analysis, and migration patterns indicated that T. marmoratus displays a weaker geographic structure than expected based on habitat distribution and physiological requirements. Demographic and statistical phylogeography analyses pointed out to a scenario of recent population expansion and high connectivity events of a more recent age than the post Last Glacial Maximum, probably associated to more humid events in Altiplano. PD of T. marmoratus populations within protected areas represents 55.6% of the total estimated PD. The unprotected populations that would contribute the most to PD are Caquena and Quebe (21%). Recent evolutionary processes and paleoclimatic changes, potentially driving shifts in habitat connectivity levels and population sizes, could explain the phylogeographic patterns recovered herein.