Anurofauna em uma área de ecótono entre Cerrado e Floresta Estacional: diversidade, distribuição e a influência de características ambientais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Lanfrediella amphicirrus gen. nov. sp. nov. Nematotaeniidae (Cestoda: Cyclophyllidea), a tapeworm parasite of Rhinella marina (Linnaeus, 1758) (Amphibia: Bufonidae)

The family Nematotaeniidae, tapeworms commonly found in the small intestines of amphibians and reptiles, includes 27 recognised species distributed among four genera: Bitegmen Jones, Cylindrotaenia Jewell, Distoichometra Dickey and Nematotaenia Lühe. The taxonomy of these cestodes is poorly defined, due in part to the difficulties of observing many anatomical traits. This study presents and describes a new genus and species of nematotaeniid parasite found in cane toads (Rhinella marina) from eastern Brazilian Amazonia. The cestodes were collected during the necropsy of 20 hosts captured in the urban area of Belém, Pará. The specimens were fixed and processed for light microscopy, scanning electron microscopy (SEM) and three-dimensional (3D) reconstruction. Samples were also collected for molecular analyses. The specimens presented a cylindrical body, two testes and paruterine organs. However, they could not be allocated to any of the four existing nematotaeniid genera due to the presence of two each of dorsal compact medullary testes, cirri, cirrus pouches, genital pores, ovaries and vitelline glands per mature segment. Lanfrediella amphicirrus gen. nov. sp. nov. is the first nematotaeniid studied using Historesin analysis, SEM and 3D reconstruction, and it is the second taxon for which molecular data have been deposited in GenBank.

Diversificação das espécies de Rhinella do grupo crucifer (Anura, Bufonidae) na Mata Atlântica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversidade, distribuição e diversificação do gênero Brachycephalus Fitzinger, 1826 (Anura: Brachycephalidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Temperature effects on baroreflex control of heart rate in the toad, Rhinella schneideri

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Orexin in the toad Rhinella schneideri: The location of orexinergic neurons and the role of orexin in ventilatory responses to hypercarbia and hypoxia

Recent reports have suggested that orexins, also known as hypocretins, play an important role in the modulation of respiratory control in mammals, but there are no data available describing the role of the orexinergic system in the peripheral and central chemoreception of non-mammalian vertebrates. Therefore, the present study was designed to examine the localization of orexin-immunoreactive neurons in the brain of toads (Rhinella schneideri) and to investigate the contribution of orexin receptor-1 (OX1R) to the hypoxic and hypercarbic ventilatory responses of these animals during light and dark phases. Our results demonstrated that the orexinergic neurons of R. schneideri are located in the suprachiasmatic nucleus of the diencephalon. Additionally, the intracerebroventricular injection of SB-334867 (OX1R selective antagonist) attenuated the ventilatory response to hypercarbia during the dark phase by acting on tidal volume and breathing frequency, while during the light phase, SB-334867 attenuated the ventilatory response to hypoxia by acting on tidal volume only. We conclude that in the toad R. schneideri, orexinergic neurons are located in the suprachiasmatic nucleus and that OX1R contributes to hypercarbic and hypoxic chemoreflexes.

Avian Use of Harvested Crop Fields in North Dakota During Spring Migration

Broad-spectrum herbicide applications and improved harvesting efficiency of crops have reduced the availability of weed seeds and waste grains for game and nongame wildlife. Over the last decade, corn and soybean plantings have steadily increased in the Prairie Pothole Region (PPR) of North Dakota, while sunflower plantings have declined. The PPR is an important corridor for migratory birds, and changes in food availabilities at stopover habitats may affect how food resources are used. In early spring 2003 and 2004, we compared bird use of harvested fields of sunflower, soybeans, small grains, and corn in the PPR of North Dakota. Across both years and all crop types, we observed 20,400 birds comprising 29 species. Flocks of Lapland Longspurs (Calcarius lapponicus) and Horned Larks (Eremophila alpestris) and flocks of Red-winged Blackbirds (Agelaius phoeniceus) made up 60% and 15%, respectively, of the bird counts. We found that species richness and bird densities were higher in harvested sunflower fields and cornfields than in harvested small-grain and soybean fields, with soybean fields harboring the fewest species and lowest bird density. Blackbird densities tended to be lower in fields tilled after fall harvest than in fields not tilled. These results suggest that some granivorous bird populations in the Northern Great Plains could be positively affected by planting of row crops with postharvest vertical structure (e.g., sunflower, corn) and use of no-till land management practices.

AN OVERVIEW OF DEPREDATING BIRD DAMAGE CONTROL IN CALIFORNIA

To many people, California is synonomous with Disneyland, freeways, Los Angeles smog, Yosemite, the California missions, or for you bird aficionados, the California Condor. But do you think about California when you eat strawberry shortcake? You should -- California leads the nation in strawberry production. How about artichokes? California produces over 98% of the artichokes raised in the United States. Dates? California produces over 99% of the dates in the United States. Yes, California is all of these, and it is much more. California may well be the most diverse state in the United States. Within its 100.2 million acres, California has the lowest place in the U.S. in Death Valley and one of the highest mountains with Mt. Whitney. Because California is such a diverse state and has a wide variety of micro- climates, it supports a uniquely diverse agriculture. Agriculture uses only about 36 million acres of its total 100.2 million acres, and most of the cash return from crops is produced on 8,6 million acres that are irrigated. California produces about 250 crops and livestock commodities (excluding nursery crops) and provides the U.S. with about 25% of its table foods. California leads the nation in the production of 46 commercial crops and livestock commodities; its farmers and ranchers marketed $8.6 billion of crop and livestock products in 1975, and the state’s harvested farm production in 1975 set a new record at 51.1 million tons. HISTORY OF BIRD PROBLEMS Records such as this are not achieved without some risk. Crops growing in Cali- fornia have always had competition from many types of vertebrate pests. The wide variety of crops grown in California and the varied climates and situations in which they are grown has resulted in many different species of birds damaging crops. Birds have compet- ed with man for his crops since the dawn of agriculture. McAtee (1932) cited examples of bird damage that occurred in a wide variety of crops in California during the early 1900s. During the 1920s, many requests for Information and relief from damage caused by a wide variety of birds, culminated in the assignment, in May 1929, of two biologists, S. E. Piper and Johnson Neff, of the former U.S. Bio- logical Survey, to initiate field studies in California. In cooperation with the Cali- fornia Department of Food and Agriculture and County Agricultural Commissioners, the study was to determine the problems and devise control procedures relative to bird depredations. Piper and Neff found such damage as Horned Larks pulling sprouting crops, House Finches disbudding deciduous fruit trees and devouring mature fruit. Blackbirds were a problem in the rice crop. Early controls were varied and, for the most part, lacked effectiveness. Flagging of fields was common to deter Horned Larks. Windmill devices were tried to frighten birds. Shooting to kill birds was common; scarecrows were.used. The six-year study brought forth the basis of most of the depredating bird control techniques still in use in California. At the end of the study, these two biologists compiled a book called “Procedure and Methods in Controlling Birds Injurious to Crops in California.” This was and still is the “Bible” for bird damage control techniques used in California. The thorough investigations conducted by these biologists resulted in techniques that have remained valid in California for over 40 years.

The breathing pattern and the ventilatory response to aquatic and aerial hypoxia and hypercarbia in the frog Pipa carvalhoi

Anuran amphibians are known to exhibit an intermittent pattern of pulmonary ventilation and to exhibit an increased ventilatory response to hypoxia and hypercarbia. However, only a few species have been studied to date. The aquatic frog Pipa carvalhoi inhabits lakes, ponds and marshes that are rich in nutrients but low in O-2. There are no studies of the respiratory pattern of this species and its ventilation during hypoxia or hypercarbia. Accordingly, the aim of the present study was to characterize the breathing pattern and the ventilatory response to aquatic and aerial hypoxia and hypercarbia in this species. With this purpose, pulmonary ventilation (V-1) was directly measured by the pneumotachograph method during normocapnic normoxia to determine the basal respiratory pattern and during aerial and aquatic hypercarbia (5% CO2) and hypoxia (5% O-2). Our data demonstrate that P. carvalhoi exhibits a periodic breathing pattern composed of single events (single breaths) of pulmonary ventilation separated by periods of apnea. The animals had an enhanced V-1 during aerial hypoxia, but not during aquatic hypoxia. This increase was strictly the result of an increase in the breathing frequency. A pronounced increase in V-1 was observed if the animals were simultaneously exposed to aerial and aquatic hypercarbia, whereas small or no ventilatory responses were observed during separately administered aerial or aquatic hypercarbia. P. carvalhoi primarily inhabits an aquatic environment. Nevertheless, it does not respond to low O-2 levels in water, although it does so in air. The observed ventilatory responses to hypercarbia may indicate that this species is similar to other anurans in possessing central chemoreceptors. (C) 2012 Elsevier Inc. All rights reserved.

Molecular phylogeny and morphometric analyses reveal deep divergence between Amazonia and Atlantic Forest species of Dendrophryniscus

Dendrophryniscus is an early diverging clade of bufonids represented by few small-bodied species distributed in Amazonia and the Atlantic Forest. We used mitochondrial (414 bp of 12S, 575 bp of 16S genes) and nuclear DNA (785 bp of RAG-1) to investigate phylogenetic relationships and the timing of diversification within the genus. These molecular data were gathered from 23 specimens from 19 populations, including eight out of the 10 nominal species of the genus as well as Rhinella boulengeri. Analyses also included sequences of representatives of 18 other bufonid genera that were publically available. We also examined morphological characters to analyze differences within Dendrophryniscus. We found deep genetic divergence between an Amazonian and an Atlantic Forest clade, dating back to Eocene. Morphological data corroborate this distinction. We thus propose to assign the Amazonian species to a new genus, Amazonella. The species currently named R. boulengeri, which has been previously assigned to the genus Rhamphophryne, is shown to be closely related to Dendrophryniscus species. Our findings illustrate cryptic trends in bufonid morphological evolution, and point to a deep history of persistence and diversification within the Amazonian and Atlantic rainforests. We discuss our results in light of available paleoecological data and the biogeographic patterns observed in other similarly distributed groups. (C) 2011 Elsevier Inc. All rights reserved.

The occurrence of defensive alkaloids in non-integumentary tissues of the Brazilian red-belly toad Melanophryniscus simplex (Bufonidae)

The red-belly toads (Melanophryniscus) of southern South America secrete defensive alkaloids from dermal granular glands. To date, all information on Melanophryniscus alkaloids has been obtained by extraction from either skins or whole organisms; however, in other amphibians, tetrodotoxins, samandarines, and bufadienolides have been detected in both skin and other organs, which raise the possibility that lipophilic alkaloids may occur in non-integumentary tissues in Melanophryniscus as well. To test this hypothesis, we studied the distribution of alkaloids in the skin, skeletal muscle, liver, and mature oocytes of the red-belly toad M. simplex from three localities in southern Brazil. Gas chromatography and mass spectrometry of skin extracts from 11 individuals of M. simplex resulted in the detection of 47 alkaloids (including isomers), 9 unclassified and 38 from 12 known structural classes. Each alkaloid that was present in the skin of an individual was also present in the same relative proportion in that individual's skeletal muscle, liver, and oocytes. The most abundant and widely distributed alkaloids were the pumiliotoxins 251D, 267C, and 323A, 5,8-disubstituted indolizidines 207A and 223D, 5,6,8-trisubstituted indolizidine 231B, 3,5-disubstituted pyrrolizidines cis-223B and cis- and trans-251K, and izidine 211C. We report the first record of piperidines in Melanophryniscus, bringing the total number of alkaloid classes detected in this genus to 16. Alkaloid composition differed significantly among the three study sites. The functional significance of defensive chemicals in non-integumentary tissues is unknown.

Antimicrobial capacity of plasma from anurans of the Atlantic Forest

The viability and interpretation of techniques for the evaluation of immunocompetence of animals in their natural environment has been largely debated. One of these methods is based on testing the antimicrobial capacity of the blood and/or plasma in vitro, which could rapidly and effectively assess the immunological conditions of natural populations. We tested the applicability of the antimicrobial capacity of plasma (ACP) assay in anuran amphibians from the Atlantic Forest. The assay was performed by measuring both the turbidity (in a spectrophotometer) and the colony forming units (CFU) of the remaining bacteria (Escherichia coli) following exposure to amphibian plasma. Although both assays were correlated, the ACP assay by spectrophotometry showed 10 times lower intra-assay variation. We also found interspecific variation in ACP, as well as the maintenance of ACP values in males from the same population, collected in different breeding seasons. Thus, the estimation of ACP by spectrophotometry provides a convenient and accurate method for evaluating innate immunocompetence in comparative and ecophysiological studies of anuran amphibians.

Novel Features of the Prenatal Horn Bud Development in Cattle (Bos taurus).

Whereas the genetic background of horn growth in cattle has been studied extensively, little is known about the morphological changes in the developing fetal horn bud. In this study we histologically analyzed the development of horn buds of bovine fetuses between ~70 and ~268 days of pregnancy and compared them with biopsies taken from the frontal skin of the same fetuses. In addition we compared the samples from the wild type (horned) fetuses with samples taken from the horn bud region of age-matched genetically hornless (polled) fetuses. In summary, the horn bud with multiple layers of vacuolated keratinocytes is histologically visible early in fetal life already at around day 70 of gestation and can be easily differentiated from the much thinner epidermis of the frontal skin. However, at the gestation day (gd) 212 the epidermis above the horn bud shows a similar morphology to the epidermis of the frontal skin and the outstanding layers of vacuolated keratinocytes have disappeared. Immature hair follicles are seen in the frontal skin at gd 115 whereas hair follicles below the horn bud are not present until gd 155. Interestingly, thick nerve bundles appear in the dermis below the horn bud at gd 115. These nerve fibers grow in size over time and are prominent shortly before birth. Prominent nerve bundles are not present in the frontal skin of wild type or in polled fetuses at any time, indicating that the horn bud is a very sensitive area. The samples from the horn bud region from polled fetuses are histologically equivalent to samples taken from the frontal skin in horned species. This is the first study that presents unique histological data on bovine prenatal horn bud differentiation at different developmental stages which creates knowledge for a better understanding of recent molecular findings.

A 1.8-kb insertion in the 3'-UTR of RXFP2 is associated with polledness in sheep

Sheep breeds show a broad spectrum of different horn phenotypes. In most modern production breeds, sheep are polled (absence of horns), whereas horns occur mainly in indigenous breeds. Previous studies mapped the responsible locus to the region of the RXFP2 gene on ovine chromosome 10. A 4-kb region of the 3'-end of RXFP2 was amplified in horned and polled animals from seven Swiss sheep breeds. Sequence analysis identified a 1833-bp genomic insertion located in the 3'-UTR region of RXFP2 present in polled animals only. An efficient PCR-based genotyping method to determine the polled genotype of individual sheep is presented. Comparative sequence analyses revealed evidence that the polled-associated insertion adds a potential antisense RNA sequence of EEF1A1 to the 3'-end of RXFP2 transcripts.

High-fidelity piezoelectric loudspeaker

This project reports on a literature review about piezoelectric loudspeakers and on an experimental research about how to improve some features of a particular horned piezoelectric tweeter. The work involves an investigation of the performance and principle of operation of piezoelectric loudspeakers to understand how the sound is generated and what its main parameters are. Also, previous research papers about how to improve this type of speakers are reported. The knowledge gained was used to reconsider and re-purpose a particular piezoelectric transducer. After characterising the original state of the device with acoustical and electroacoustical measurements, some improvements were implemented. Moreover, interesting conclusions were reached based on the results of the tests that were carried out. A structural study with a scanning laser was then completed. These sections demonstrated the need for providing the speaker with a rear suspension that guides the vibration of the membrane. Finally, an inverse filter was designed in order to get a flat output response. After simulating the results with Matlab, validating experiments were run in the anechoic chamber with great success. RESUMEN. Este proyecto consta de un estudio detallado sobre piezoelectricidad y altavoces piezoeléctricos, así como de una parte experimental consistente en mejorar algunas características de un altavoz piezoeléctrico particular: un tweeter de bocina. El estudio profundiza en cuáles son los principios de funcionamiento y los principales parámetros de este tipo de altavoces. Con el conocimiento adquirido a partir de trabajos de previos sobre el tema e investigación bibliográfica se ha llevado a cabo la parte experimental. Esta parte ha requerido de una serie de medidas acústicas y electroacústicas para, primero, caracterizar el altavoz en su estado original y para posteriormente buscar y validar posibles mejoras, principalmente en la respuesta en frecuencia. Además, se ha realizado un estudio estructural del diafragma a partir de medidas tomadas con un vibrómetro laser Doppler. De estos tres procesos se concluyó que el altavoz bajo estudio tiene un problema en el soporte del cristal piezoeléctrico y se demostró la necesidad de equipar el dispositivo de una suspensión trasera que controle el movimiento del diafragma. Finalmente, se ha diseñado e implementado en Matlab un filtro inverso, con el objetivo de conseguir una respuesta plana a la salida del altavoz. Su funcionamiento fue validado en la cámara anecoica satisfactoriamente.