Integrated multiphase ecological risk assessment of heavy metals for migratory water birds in wetland ecosystem: A case study of Dongzhangwu Wetland, China.

Wetland is an important feature of the landscape that provides beneficial services. However, with the ever-increasing heavy metal load, the quality of wetlands is deteriorating. Dongzhangwu Wetland in Hebei, China, was taken as our study site. It provides breeding and foraging grounds to migratory water birds such as Little Egret (Egretta garzetta), Great Egret (Ardea alba), and Grey Heron (Ardea cinerea). The current study aimed to quantify heavy metals exposure hazard and risk to the water migratory birds by employing a non-destructive approach. Oral intake was considered the main exposure route to calculate total exposure via multiple phases. The concentrations of Cr, Zn, Cu, Pb, As, Ni, Mn, and Cd were investigated in water, soil and food compartments of three different habitat components (Longhe River, Natural Pond and Fish Pond). The results showed: (1) The trend of potential daily dose (PDD) was Mn > Zn > Cr > Pb > Ni > Cu > As > Cd while for hazard quotient (HQ) was Cr > Pb > Cu > Zn > As > Ni > Mn > Cd, making Cr, Pb, Cu, Zn, and As the priority pollutants in all habitats, with Natural Pond eliciting the highest exposure. (2) The cumulative heavy metal exposure, explained by the integrated nemerow risk index, characterized all three habitats for all the birds in high exposure risk category. (3) The exposure frequency index identified that all the birds are frequently exposed to heavy metals from multiple phases in all three habitats. (4) Little Egret is at the highest pedagogy of exposure from single or multiple heavy metal(s) in all three habitats. A rigorous management plan for identified priority pollutants is required to improve wetland functioning and ecological services. Decision-makers could use the developed tissue residue objectives for protecting Egret species in Dongzhangwu Wetland as benchmarks.

Displasia Fibrosa tratada com Técnica de Masquelet: relato de caso / Fibrous Dysplasia treated with Masquelet Technique: case report

Introdução: A displasia fibrosa (DF) do osso é uma desordem congênita, rara, que corresponde de 5 a 10% dos tumores ósseos benignos, não hereditária, que cursa com amplo espectro de apresentação, variando do assintomático à dor óssea, fraturas de repetição, deformidades ósseas (fêmur em cajado de pastor e fácies leonina) e compressão de nervos cranianos. Histologicamente é composta de estroma fibroso celular de baixo a moderado grau circundando trabéculas ósseas de formato irregular sem borda osteoblástica. Todos os casos contêm a mutação GNAS1. A DF apresenta duas formas: a monostótica, mais comum (70-80%), e a poliostótica, mais rara (20-30%), que quando acompanhada de manchas café-com-leite e puberdade precoce constitui a síndrome de McCune-Albright ou Síndrome de Mazabraud em casos mais raros. O tratamento pode ser feito com medicamentos como bifosfonato ou de forma cirúrgica, objetivando-se a correção das lesões com curetagem e enxertia óssea ou como iremos mostrar a seguir, pela Técnica de Masquelet. Este trabalho relata o caso de um menino de 20 anos de idade cujos sinais e sintomas conduziam ao diagnóstico de DF sendo realizado tratamento com Técnica de Masquelet e follow up de 18 meses. Além disso, faz revisão de literatura sobre uma doença pouco comum, com variada gama de diagnósticos diferenciais. Objetivo: relatar um caso de displasia fibrosa com tratamento cirúrgico de enxerto autólogo de fíbula pela Técnica de Masquelet. Método: relato de caso de paciente do Ambulatório de Especialidade do Hospital do Servidor Público Municipal, de 20 anos de idade, que foi acompanhado por 1 ano e meio apresentando um tumor ósseo na tíbia compatível com diagnóstico de displasia fibrosa, que ao longo desse período foi submetido à Técnica de Masquelet. Conclusão: É pouco descrito na literatura o tratamento de displasia fibrosa pela Técnica de Masquelet, que mostrou ter ótimo resultado funcional para o paciente estudado. Palavras-chave: Displasia Fibrosa Óssea. Displasia Fibrosa. Técnica de Masquelet. Técnica de Membrana Induzida.

Interaction between live vaccines for coccidiosis and phytogenic compounds in the diet of broilers.

This study aimed to determine whether live vaccines alone or associated with phytogenic compounds supplied via feed could improve performance, intestinal health, serum biochemistry, and coccidiostatic action in broilers up to 42 days of age, challenged with Eimeria spp., compared with a traditional anticoccidial program. We used 800 birds divided into five treatments of eight repetitions each (n = 20): NC - Negative control (absence of additives in the feed and without coccidian challenge); PC - Positive control (absence of additives and with coccidian challenge at 21 days); AAP - Anticoccidial additives program, including salinomycin and nicarbazine, with coccidian challenge at 21 days of life; VAC - Vaccination on day 1 of life (Hipracox HIPRA®) against coccidiosis and subsequent coccidian challenge at 21 days; VAC + BPC - Vaccination (day 1 of life), the addition of 200 ppm of blend phytogenic compound (BPC) based on carvacrol and cinnamaldehyde to the diet, with coccidian challenge at 21 days of life. The birds in the AAP group more significant weight gain and greater body weight at 21 days than the NC and 42 days of age than the NC and VAC + BPC groups. The excretion of Eimeria oocysts in the excreta of birds was higher in PC than in NC, and in the litter, PC had higher counts than NC, AAP, and VAC + BPC. A higher mean score of intestinal lesions for Eimeria acervulina was observed in VAC and VAC + BPC birds, while for Eimeria maxima, the highest score was found in PC. Higher serum levels of total protein due to the increase in globulins were observed in birds of AAP and VAC than NC. Our findings suggest that vaccination for coccidiosis shortly after the birth of chicks allows performance similar to the conventional coccidiosis control program via diet.

Quaternary climatic fluctuations influence the demographic history of two species of sky-island endemic amphibians in the Neotropics.

We evaluated the role of Quaternary climatic fluctuations on the demographic history and population structure of amphibian species endemic to the 'campo rupestre' in the Neotropics, evaluating their distributional shifts, demographic changes, and lineage formation from the end of Pleistocene to present. We chose two anurans endemic to the high-elevation 'campo rupestre' in the Espinhaço Range (ER) in northeastern and southeastern Brazil (Bokermannohyla alvarengai and Bokermannohyla oxente), as models to test the role of Quaternary climatic fluctuations over their distribution range in this region. We collected tissue samples throughout their distribution range and used statistical phylogeography to examine processes of divergence and population demography. We generated spatial-temporal reconstructions using Bayesian inference in a coalescent framework in combination with hind-cast projections of species distribution models (SDMs). We also used the results and literature information to test alternative diversification scenarios via approximate Bayesian computation (ABC). Our results show that Quaternary climatic fluctuations influenced the geographic ranges of both species showing population expansion during the last glacial maximum (LGM) and range contraction during interglacial periods, as inferred from selected ABC models and from past projections of SDMs. We recovered Pleistocene diversification for both species occuring in distinctly unique periods for each taxon. An older and range-restricted lineage was recovered in a geographically isolated geological massif, deserving conservation and further taxonomic study. The diversification and distribution of these amphibian species endemic to the Neotropical 'campo rupestre' were influenced by Quaternary climatic fluctuations. The expansion of cold adapted species restricted to higher elevations during glacial periods and their concomitant retraction during interglacial periods may have been crucial for producing patterns of species richness and endemism along elevation gradients in tropical and subtropical domains. Such processes may influence the evolution of the biota distributed in heterogeneous landscapes with varied topography.

Biallelic UBE4A loss-of-function variants cause intellectual disability and global developmental delay.

PURPOSE: To identify novel genes associated with intellectual disability (ID) in four unrelated families. METHODS: Here, through exome sequencing and international collaboration, we report eight individuals from four unrelated families of diverse geographic origin with biallelic loss-of-function variants in UBE4A. RESULTS: Eight evaluated individuals presented with syndromic intellectual disability and global developmental delay. Other clinical features included hypotonia, short stature, seizures, and behavior disorder. Characteristic features were appreciated in some individuals but not all; in some cases, features became more apparent with age. We demonstrated that UBE4A loss-of-function variants reduced RNA expression and protein levels in clinical samples. Mice generated to mimic patient-specific Ube4a loss-of-function variant exhibited muscular and neurological/behavioral abnormalities, some of which are suggestive of the clinical abnormalities seen in the affected individuals. CONCLUSION: These data indicate that biallelic loss-of-function variants in UBE4A cause a novel intellectual disability syndrome, suggesting that UBE4A enzyme activity is required for normal development and neurological function.

Phytogenic blend protective effects against microbes but affects health and production in broilers.

The aim of this study was to determine whether addition of a phytogenic blend in the feed of broilers to replace conventional antimicrobials as a performance enhancer would improve or maintain productive efficiency. The phytogenic blend was based on curcuminoids, cinnamaldehyde and glycerol monolaurate. We used 480 birds divided into three groups with eight repetitions per group and 20 birds per repetition. The groups were identified as antimicrobial-treated: basal feed with antibiotics and coccidiostatic agents; phytogenic blend: basal feed with blend; and control, only basal feed. Zootechnical performance was measured on days from 1 to 42, with body weight measured at days 1, 7, 21 and 42. We collected excreta for parasitological analysis and total bacterial counts to determine if the phytogenic blend had kept the bacteria and coccidia in counts smaller or similar to that resulting from use of conventional performance enhancer. Other variables were also measured to complement our research, i.e., if the consumption of bend is good for the health of the birds (without causing toxicity and negatively altering the metabolism and intestinal morphometry) and does not interfere in the quality of the meat. Because the bacteria are often opportunistic, we challenged all birds at 23 days of age with high doses of oral oocysts (28,000 oocysts). Birds supplemented with the blend showed inferior performance compared to birds in the control and antimicrobial treated group (P < 0.05). We found a smaller number of oocysts of Eimeria spp. in the excreta at 42 days in the treatment with blend and antimicrobial treated group (P < 0.05). In terms of total bacterial counts, there were lower counts in the birds of the blend group than in the control group (P < 0.05). The blend increased the yellow intensity and the luminosity of the meat (P < 0.05), as well as cooking weight losses (P < 0.05) compared those of the control. We observed higher total levels of saturated fatty acids in meat from the blend and antimicrobial treated group (P < 0.05), as well as lower levels of monounsaturated fatty acids in the blend group (P < 0.05). The inclusion of a phytogenic blend to replace conventional antimicrobials and anticoccidial agents in the diet of chickens was able to control bacteria as well as coccidia; however, it ends up harming health and production.

Microencapsulated carvacrol and cinnamaldehyde replace growth-promoting antibiotics: Effect on performance and meat quality in broiler chickens.

The aim was to evaluate the use of mixture of microencapsulated carvacrol and cinnamaldehyde as a replacement for growth-promoting antibiotics in broiler diets on performance, intestinal quality, organ development, carcass yields and cuts, and meat quality. In the trial were used 600 male chicks, allocated in a completely randomized design, with five treatments and eight replicates of 15 birds, reared up to 41 days of age. The treatments were: Negative Control (NC), Positive Control (PC) 30 mg/kg of virginiamycin, NC+100 mg/kg of essential oils, NC+200 mg/kg of essential oils and NC+400 mg/kg of essential oils. Essential oils were composed by a micro-encapsulated blend, with of 60% cinnamaldehyde, 30% carvacrol and 10% carrier. Birds received essential oils achieved performance equivalent to those birds received PC diets, having better development than NC broilers. No differences were found on relative organ weight, intestinal mucosa and meat quality parameters, however, higher villus:cript ratio was found in PC, NC+200 and NC+400 groups. Meat crude protein and yellowness were influenced by inclusion of carvacrol and cinnamaldehyde. It was concluded microencapsulated carvacrol and cinnamaldehyde can replace growth-promoting antibiotic in broiler diets, ensuring performance, intestinal integrity and broiler meat quality.

Sarcomere Length Nonuniformity and Force Regulation in Myofibrils and Sarcomeres.

The smallest contractile unit in striated muscles is the sarcomere. Although some of the classic features of contraction assume a uniform behavior of sarcomeres within myofibrils, the occurrence of sarcomere length nonuniformities has been well recognized for years, but it is yet not well understood. In the past years, there has been a great advance in experiments using isolated myofibrils and sarcomeres that has allowed scientists to directly evaluate sarcomere length nonuniformity. This review will focus on studies conducted with these preparations to develop the hypotheses that 1) force production in myofibrils is largely altered and regulated by intersarcomere dynamics and that 2) the mechanical work of one sarcomere in a myofibril is transmitted to other sarcomeres in series. We evaluated studies looking into myofibril activation, relaxation, and force changes produced during activation. We conclude that force production in myofibrils is largely regulated by intersarcomere dynamics, which arises from the cooperative work of the contractile and elastic elements within a myofibril.

Sarcomere length non-uniformities dictate force production along the descending limb of the force-length relation.

The force-length relation is one of the most defining features of muscle contraction, and yet a topic of debate in the literature. The sliding filament theory predicts that the force produced by muscle fibres is proportional to the degree of overlap between myosin and actin filaments, producing a linear descending limb of the active force-length relation. However, several studies have shown forces that are larger than predicted, especially at long sarcomere lengths (SLs). Studies have been conducted with muscle fibres, preparations containing thousands of sarcomeres that make measurements of individual SL challenging. The aim of this study was to evaluate force production and sarcomere dynamics in isolated myofibrils and single sarcomeres from the rabbit psoas muscle to enhance our understanding of the theoretically predicted force-length relation. Contractions at varying SLs along the plateau (SL = 2.25-2.39 µm) and the descending limb (SL > 2.39 µm) of the force-length relation were induced in sarcomeres and myofibrils, and different modes of force measurements were used. Our results show that when forces are measured in single sarcomeres, the experimental force-length relation follows theoretical predictions. When forces are measured in myofibrils with large SL dispersions, there is an extension of the plateau and forces elevated above the predicted levels along the descending limb. We also found an increase in SL non-uniformity and slowed rates of force production at long lengths in myofibrils but not in single sarcomere preparations. We conclude that the deviation of the descending limb of the force-length relation is correlated with the degree of SL non-uniformity and slowed force development.

Unraveling the species diversity and relationships in the Leptodactylus mystaceus complex (Anura: Leptodactylidae), with the description of three new Brazilian species.

Members of the Leptodactylus mystaceus species complex are widely distributed in forests and open formations of South America east of the Andes. Species of the complex are morphologically similar or indistinguishable among each other, but acoustic data have been the cornerstone for species discrimination across their geographic ranges. In this paper, we re-examine the monophyly, species diversity, and relationships in the L. mystaceus complex on the basis of morphology, coloration, acoustics, and DNA sequences. Morphological and color patterns originally used to the allocation of species to the L. mystaceus complex are also reassessed. Our results revealed three new species from the Brazilian Atlantic Forest and Cerrado, which are named and described herein, based mainly on acoustic and molecular data. Populations assigned to the lineage widely distributed across the South American Dry Diagonal (DD), reported in this study as L. cf. mystaceus, is likely paraphyletic with respect to the nominal species (Amazonian lineage), but additional data are still needed to address the taxonomic status of the DD lineage.

Amphibians from Serra do Cipó, Minas Gerais, Brasil. VI: A New Species of the Physalemus deimaticus Group (Anura, Leptodactylidae).

We describe a new species of Physalaemus assigned to the Physalaemus signifer Clade, and it is morphologically similar to P. rupestris, from the highlands of the Serra do Cipó in the southern Espinhaço Range, State of Minas Gerais, Southeastern Brazil. The new species is diagnosed by using the following combination of character states: presence of an arrow-shaped blotch on the dorsum of body; presence of a median longitudinal light stripe over urostyle region; belly and ventral surface of thigh marbled with dark gray irregularly shaped blotches on a bluish background; absence of reddish coloration over axillary and inguinal regions in live individuals; bright orange dots scattered over head, upper lip, and dorsum of body in live individuals; ventral surface of hand and foot red in live individuals; small size (adult male SVL=16.2-18.2 mm); presence of brown, not divided, nuptial pad in males; END/ED in males ranging from 0.85-0.93; supernumerary tubercles on foot absent; tarsal fold absent; tarsal tubercle absent; texture of posterior region of belly and ventral surface of thigh smooth; advertisement call composed of two note types (note A + B); advertisement call duration of 0.80-1.28 s; note A with ascending amplitude until mid-note then descending towards the end of the note; note B with pulses arranged in 5-7 groups; and dominant frequency of note A from 1734.4-2765.6 Hz and of note B from 1507.3-2859.4 Hz. A phylogenetic analysis based on mitochondrial DNA sequences recovered the new species nested within the Physalaemus deimaticus species group. Additionally, we redescribe the call of Physalaemus rupestris and provide a review of the geographic distribution and conservation status of the species belonging to the P. deimaticus species group.

Different gene expression profiles in iPSC-derived motor neurons from ALS8 patients with variable clinical courses suggest mitigating pathways for neurodegeneration.

Amyotrophic lateral sclerosis type 8 (ALS8) is an autosomal dominant form of ALS, which is caused by pathogenic variants in the VAPB gene. Here we investigated five ALS8 patients, classified as 'severe' and 'mild' from a gigantic Brazilian kindred, carrying the same VAPB mutation but displaying different clinical courses. Copy number variation and whole exome sequencing analyses in such individuals ruled out previously described genetic modifiers of pathogenicity. After deriving induced pluripotent stem cells (iPSCs) for each patient (N = 5) and controls (N = 3), motor neurons were differentiated, and high-throughput RNA-Seq gene expression measurements were performed. Functional cell death and oxidative metabolism assays were also carried out in patients' iPSC-derived motor neurons. The degree of cell death and mitochondrial oxidative metabolism were similar in iPSC-derived motor neurons from mild patients and controls and were distinct from those of severe patients. Similar findings were obtained when RNA-Seq from such cells was performed. Overall, 43 genes were upregulated and 66 downregulated in the two mild ALS8 patients when compared with severe ALS8 individuals and controls. Interestingly, significantly enriched pathways found among differentially expressed genes, such as protein translation and protein targeting to the endoplasmic reticulum (ER), are known to be associated with neurodegenerative processes. Taken together, the mitigating mechanisms here presented appear to maintain motor neuron survival by keeping translational activity and protein targeting to the ER in such cells. As ALS8 physiopathology has been associated with proteostasis mechanisms in ER-mitochondria contact sites, such differentially expressed genes appear to relate to the bypass of VAPB deficiency.

A grave drum at rocky springs: the vocalization of Bokermannohyla alvarengai (Anura: Hylidae) from the Espinhaço Range, southeastern Brazil, with notes on its natural history.

Communication in anurans is mediated mainly by acoustic signals, which are relevant for mating, territoriality, and other social interactions (Wells 2007). Although these signals are usually conspicuous and relatively stereotyped, some Neotropical frogs display noticeable diversity and complexity in their calls. For instance, Bokermannohyla, a genus of gladiator treefrog from the Atlantic Forest, Cerrado, and Caatinga Biomes of Brazil, often have complex vocal repertoires (Carvalho et al. 2012; Nali Prado 2014), whilst some congeners have simple and stereotyped calls emitted continuously (Carvalho et al. 2013; Taucce et al. 2015; Giaretta et al. 2016; Rocha et al. 2016).

The load dependence and the force-velocity relation in intact myosin filaments from skeletal and smooth muscles.

In the present study we evaluated the load dependence of force produced by isolated muscle myosin filaments interacting with fluorescently labeled actin filaments, using for the first time whole native myosin filaments. We used a newly developed approach that allowed the use of physiological levels of ATP. Single filaments composed of either skeletal or smooth muscle myosin and single filaments of actin were attached between pairs of nano-fabricated cantilevers of known stiffness. The filaments were brought into contact to produce force, which caused sliding of the actin filaments over the myosin filaments. We applied load to the system by either pushing or pulling the filaments during interactions and observed that increasing the load increased the force produced by myosin and decreasing the load decreased the force. We also performed additional experiments in which we clamped the filaments at predetermined levels of force, which caused the filaments to slide to adjust the different loads, allowing us to measure the velocity of length changes to construct a force-velocity relation. Force values were in the range observed previously with myosin filaments and molecules. The force-velocity curves for skeletal and smooth muscle myosins resembled the relations observed for muscle fibers. The technique can be used to investigate many issues of interest and debate in the field of muscle biophysics.

Anurans of the Caparaó National Park and surroundings, southeast Brazil / Anuros do Parque Nacional do Caparaó e arredores, sudeste brasileiro

Abstract The Atlantic Forest (AF) is one of the biodiversity hotspots of the world, and the most fragmented biome of Brazil. This biome includes different phytophysiognomies, as riparian, slope, cloudy forests, and grasslands. Such complexity, allied to huge latitudinal and high elevational range, provides diverse habitats and conditions for amphibian speciation. As a result, there are over 600 amphibian species known to occur in the AF. Within this biome the Caparaó National Park (CNP) is relevant, as it includes the highest peak of the biome, the Pico da Bandeira at almost 3,000 m above sea level, as well as different phytophysiognomies as rocky fields and humid forests. In spite of that, its amphibian fauna is still poorly described. We inventoried amphibians at the CNP and surrounding areas from 2016 to 2018 and recorded 47 anuran species, of which two are locally endemic and at least six have not been described yet. Additionally, we compiled data from previous surveys (2004 to 2008) and secondary data from scientific collections. All together, we registered a total of 61 anuran species from 12 families for the CNP and surroundings, placing this area among the 10 amphibian richest sites in the AF. Some of these species are represented by only one or two collected specimens and have not been registered in the CNP since the 1980's, such as Thoropa lutzi and Hylodes vanzolinii. These species could be examples of population declines or even past local extinctions, highlighting the need of further sampling efforts in that highly biodiverse site.

Resumo A Mata Atlântica (MA) é considerada um dos hotspots de biodiversidade do mundo, sendo o bioma mais degradado no Brasil. Este bioma compreende muitas fitofisionomias, como floresta ombrófila, semidecidual, e campos de altitude. Esta complexidade, aliada à enorme variação latitudinal e altitudinal, disponibiliza habitats e condições diversas à especiação dos anfíbios. Como resultado, atualmente são conhecidas mais de 600 espécies de anfíbios que ocorrem na MA. Dentro deste bioma o Parque Nacional do Caparaó (PNC) é relevante, pois inclui o ponto mais alto da Mata Atlântica, o Pico da Bandeira com cerca de 3000 m de elevação, assim como diferentes fitofisionomias, como campos rupestres e florestas úmidas. Apesar disso, sua fauna de anfíbios é ainda pobremente descrita. Realizamos inventários de anfíbios no PNC e entorno de 2016 a 2018 e registramos 47 espécies de anuros, das quais duas são endêmicas e pelo menos seis ainda não foram descritas. Adicionalmente, compilamos dados de coletas anteriores (2004 a 2008) e dados secundários de coleções científicas. Ao juntar todos os dados registramos um total de 61 espécies de anuros de 12 famílias para o PNC e arredores, colocando esta área entre as 10 áreas mais ricas em anfíbios da MA. Algumas destas espécies estão representadas apenas por um ou dois exemplares em coleções e não foram registradas no PNC desde a década de 1980, como Thoropa lutzi e Hylodes vanzolinii. Estas espécies podem ser exemplos de declínios populacionais ou mesmo de extinções locais, destacando a necessidade de maiores esforços amostrais neste local altamente biodiverso.

The tadpole of Physalaemus orophilus from the Atlantic rainforest of southeastern Brazil (Amphibia, Anura, Leptodactylidae).

The neotropical genus Physalaemus Fitzinger is currently composed by 48 species (Cardozo Pereyra 2018). Recently, a phylogenetic analysis aiming to investigate the internal relationships of the genus recovered two major clades, the Physalaemus cuvieri and P. signifer clades (Lourenço et al. 2015). The following species groups were retrieved in the first clade: P. biligonigerus, P. cuvieri, P. gracilis, P. henselii, and P. olfersii groups (Lourenço et al. 2015). This proposal redefined the P. olfersii group including P. olfersii (Lichtenstein Martens), P. soaresi Izecksohn, P. maximus Feio, Pombal, Caramaschi, P. feioi Cassini, Cruz, Caramaschi and P. lateristriga (Steindachner). The authors also allocated tentatively P. orophilus Cassini, Cruz, Caramaschi, and P. insperatus Cruz, Cassini, Caramaschi in the P. olfersii group due to their morphological similarity with the other species (Cruz et al. 2008; Cassini et al. 2010). Otherwise, Physalaemus aguirrei Bokermann was not recovered nested within this group, contradicting what was suggested in a previous phenetic analysis (Nascimento et al. 2005). Members of the P. olfersii group inhabit the Atlantic rainforest and most of them have a similar advertisement calls with pulsed notes, without frequency modulation and harmonic structure (Giaretta et al. 2009; Cassini et al. 2010; Lourenço et al. 2015). Regarding their larval stage, only P. soaresi, P. maximus, and P. olfersii have their tadpoles described (Weber et al. 2005; Baêta et al. 2007; Giaretta et al. 2009). Physalaemus orophilus occurs in montane Atlantic Forest sites at the eastern slope of the Espinhaço Range in the State of Minas Gerais, southeastern Brazil (Cassini et al. 2010). Herein, we describe the tadpole of P. orophilus from Quadrilátero Ferrífero mountain region, southern limit of the Espinhaço Range and compared it to the known tadpoles of the P. olfersii group.

Notes on vocalizations of Brazilian amphibians IV: advertisement calls of 20 Atlantic Forest frog species.

Bioacoustics is a powerful tool used for anuran species diagnoses, given that advertisement calls are signals related to specific recognition and mate attraction. Thus, call descriptions can support species taxonomy. In spite of that, call descriptions are lacking for many species, delaying advances in biodiversity research. Here, we describe the advertisement calls of 20 anuran species from the Brazilian Atlantic Forest. We accessed 50 digital recordings deposited in the Fonoteca Neotropical Jacques Vielliard. Acoustic analyses were carried out in the software Raven pro 1.5. We provide a general comparison of call structure among species inside taxonomic groups and genera. The vocalizations described here belong to poorly known species, which are representatives of six families: Brachycephalidae, Bufonidae, Ceratophryidae, Cycloramphidae, Hylidae, and Phyllomedusidae. Despite this, still there are 163 species of anurans from Atlantic Forest with calls not formally described. Our work represents an important step in providing data for a taxonomic perspective and improving the knowledge of the Atlantic Forest anuran diversity.

Modelling Highly Biodiverse Areas in Brazil.

Traditional conservation techniques for mapping highly biodiverse areas assume there to be satisfactory knowledge about the geographic distribution of biodiversity. There are, however, large gaps in biological sampling and hence knowledge shortfalls. This problem is even more pronounced in the tropics. Indeed, the use of only a few taxonomic groups or environmental surrogates for modelling biodiversity is not viable in mega-diverse countries, such as Brazil. To overcome these limitations, we developed a comprehensive spatial model that includes phylogenetic information and other several biodiversity dimensions aimed at mapping areas with high relevance for biodiversity conservation. Our model applies a genetic algorithm tool for identifying the smallest possible region within a unique biota that contains the most number of species and phylogenetic diversity, as well as the highest endemicity and phylogenetic endemism. The model successfully pinpoints small highly biodiverse areas alongside regions with knowledge shortfalls where further sampling should be conducted. Our results suggest that conservation strategies should consider several taxonomic groups, the multiple dimensions of biodiversity, and associated sampling uncertainties.

Almost a hundred years later, the advertisement call of Aparasphenodon brunoi Miranda-Ribeiro 1920 (Anura: Hylidae) from the Atlantic Forest.

The Neotropical genus Aparasphenodon Miranda-Ribeiro is composed of five recognized species commonly referred as "casque-headed frogs" (Frost 2018). Aparasphenodon brunoi Miranda-Ribeiro occurs in Atlantic Forest lowlands from the states of São Paulo to Bahia, Brazil (Feio et al. 1998; Mollo Neto Teixeira Jr. 2012; Ruas et al. 2013). The biology of most Aparasphenodon species is poorly known. Within the genus, until now only Aparasphenodon arapapa Pimenta, Napoli Haddad has its call formally described (Lourenço-de-Moraes et al., 2013, Forti et al., 2018, Guerra et al., 2018). Herein, we describe the advertisement call of Aparasphenodon brunoi Miranda-Ribeiro.

Past vicariance promoting deep genetic divergence in an endemic frog species of the Espinhaço Range in Brazil: The historical biogeography of Bokermannohyla saxicola (Hylidae).

The highland endemic frog Bokermannohyla saxicola occurs within the Espinhaço Range, the most extensive and continuous orogenic belt of the Brazilian territory, located in southeastern Brazil. We used mitochondrial DNA markers to test for spatial structure, to investigate the likely influence of past vicariant events, to evaluate demographic dynamics along the species range, and to understand the role of habitat discontinuities in promoting connectivity and diversity along the range. We found four major monophyletic lineages, each one associated with distinct mountain tops. The divergence time found between the four main clades clearly pre-dated the Pleistocene, except for the most recent separation. We observed no signs of population expansion for most of the sampling sites along the range, and a higher genetic diversity in the most continuous and central highland plateau, compared to smaller marginal regions. The Espinhaço Range harbors four deeply divergent lineages of B. saxicola within areas restricted by barriers for millions of years. These relatively isolated populations were kept apart by discontinuities represented by lowland habitats between mountain tops. Most of the lineage divergences occurred earlier than the Pleistocene, thus they cannot be solely explained by climatic oscillations of this epoch. However, within-lineage divergence times were all dated from the Pleistocene, suggesting an important effect in population dynamics. We also suggest that some marginal populations like those from Serra Negra and Serra de Itacambira can be the result of recent colonization events. Finally, in the southern Espinhaço region, the most continuous central highland area shows greater genetic diversity than the marginal discontinuous areas, where we have also observed a significant correlation between genetic and geographic distances. Bokermannohyla saxicola is a good model to study the biogeography of the Espinhaço Range because its high genetic structure reflects ancient as well as recent geological/climatic events, with important implications for conservation.