ABSTRACT
Bufonid parotoid macrogland secretion contains several low molecular mass molecules, such as alkaloids and steroids. Nevertheless, its protein content is poorly understood. Herein, we applied a sample preparation methodology that allows the analysis of viscous matrices in order to examine its proteins. Methods: Duttaphrynus melanostictus parotoid macrogland secretion was submitted to ion-exchange batch sample preparation, yielding two fractions: salt-displaced fraction and acid-displaced fraction. Each sample was then fractionated by anionic-exchange chromatography, followed by in-solution proteomic analysis. Results: Forty-two proteins could be identified, such as acyl-CoA-binding protein, alcohol dehydrogenase, calmodulin, galectin and histone. Moreover, de novo analyses yielded 153 peptides, whereas BLAST analyses corroborated some of the proteomic-identified proteins. Furthermore, the de novo peptide analyses indicate the presence of proteins related to apoptosis, cellular structure, catalysis and transport processes. Conclusions: Proper sample preparation allowed the proteomic and de novo identification of different proteins in the D. melanostictus parotoid macrogland secretion. These results may increase the knowledge about the universe of molecules that compose amphibian skin secretion, as well as to understand their biological/physiological role in the granular gland.(AU)
Subject(s)
Animals , Steroids , Bufonidae/parasitology , Proteomics , AlkaloidsABSTRACT
Abstract Although a group of soft ticks (Argasidae) associated with amphibians was recently discovered in Brazilian rainforests, parasitism by these ticks on cold-blooded animals remains less common than on mammal and bird species. In this study, we identified ticks that were collected from toads that had been caught in December 2016 and January 2017, at Itinguçú waterfall (22°54'05" S; 43°53'30" W) in the municipality of Itaguaí, state of Rio de Janeiro. Tick specimens were identified using a morphological and molecular approach. In total, twelve larvae of Ornithodoros ticks were collected from three individuals of Rhinella ornata and were identified as Ornithodoros faccinii. Our results include a longer 16S rRNA mitochondrial sequence for O. faccinii that supports its phylogenetic relatedness to Ornithodoros saraivai, and we report this tick species parasitizing Rhinella toads for the first time in Brazil.
Resumo Embora um grupo de carrapatos moles (Argasidae) associado a anfíbios tenha sido recentemente descoberto nas florestas brasileiras, o parasitismo por esses carrapatos em animais de sangue frio permanece menos comum do que nas espécies de mamíferos e aves. Neste estudo, identificamos carrapatos que foram coletados de sapos capturados em dezembro de 2016 e janeiro de 2017, na cachoeira de Itinguçú (22°54'05" S; 43°53'30" W) no município de Itaguaí, estado do Rio de Janeiro. Os espécimes de carrapatos foram identificados usando uma abordagem morfológica e molecular. No total, doze larvas de carrapatos Ornithodoros foram coletadas de três indivíduos de Rhinella ornata e foram identificadas como Ornithodoros faccinii. Nossos resultados incluem uma maior seqüência mitocondrial 16S rRNA para O. faccinii que suporta sua relação filogenética com Ornithodoros saraivai e relatamos esta espécie de carrapato parasitando sapos Rhinella pela primeira vez no Brasil.
Subject(s)
Animals , Tick Infestations/parasitology , Bufonidae/parasitology , Ornithodoros/genetics , Phylogeny , Brazil , RNA, Ribosomal, 16S , Ornithodoros/anatomy & histology , Ornithodoros/classificationABSTRACT
We sampled ticks from specimens of the rococo toad Rhinella schneideriby flannel dragging on two Islands located in the São Francisco River near the Três Marias hydroelectric dam, southeastern Brazil. A total of 120 toads was examined, of which 63 (52.5%) were parasitized only by Amblyomma rotundatumtotaling 96 larvae, 163 nymphs and 134 females. The burden of parasitism ranged from one to 43 ticks, with a mean intensity of infestation of 6.2±5.5 ticks per host. The tick A. rotundatumexhibited highly aggregated distribution. Peak abundance of larvae and nymphs occurred in the dry season (May to September), whereas peak abundance of females occurred in the wet season (October to April). We collected most ticks near the head and hind limbs of R. schneideri. The finding of two engorged A. rotundatumnymphs in the same resting places of two toads and the absence of this species in the dragged areas suggest a nidicolous behavior at the studied site.
Sapos da espécie Rhinella schneideri foram capturados e examinados para coleta das fases parasitárias, assim como arrasto de flanela para coleta das fases de vida livres de carrapatos em duas ilhas localizadas no rio São Francisco , próximas à represa Três Marias, região sudeste do Brasil. No total, 120 indivíduos foram examinados, dos quais 63 (52,5%) estavam parasitados por Amblyomma rotundatum totalizando 96 larvas, 163 ninfas e 134 fêmeas. A abundância do parasitismo variou de 1 a 43 carrapatos, com uma intensidade média de infestação de 6,2±5,5 carrapatos/hospedeiro. A infestação por A. rotundatumapresentou uma distribuição altamente agregada. O pico de abundância de larvas e ninfas ocorreu na estação seca (maio a setembro), enquanto o pico de abundância de fêmeas ocorreu na estação chuvosa (outubro a abril). A maioria dos carrapatos foi coletada na região da cabeça e membros posteriores. A presença de duas ninfas ingurgitadas de A. rotundatum nos mesmos lugares de descanso de dois sapos e a ausência desta espécie na coleta por arrasto de flanela sugere um comportamento nidicola no local estudado.
Subject(s)
Animals , Bufonidae/parasitology , Host-Parasite Interactions , Tick Infestations/veterinary , Parasite Load/veterinary , Parasitic Diseases/parasitology , Hemorrhage/veterinary , Weight LossABSTRACT
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.
Subject(s)
Animals , Female , Male , Bufonidae/parasitology , Cestoda/classification , Brazil , Cestoda/anatomy & histology , Cestoda/ultrastructure , Imaging, Three-Dimensional , Microscopy, Electron, ScanningABSTRACT
A total of 108 toads and frogs were collected by hand from January to August 1994 from different sites in the southwest of Iran, mainly from Khouzestan province. Among 79 green toads [Bufo viridis], 93.7% were infected with one or more parasite species. There was no significant difference between snout-ventral lengths [SVL] of infected [n = 74] and uninfected [n = 5] toads [P = 0.323; Student's t-test], also between the rate of infection in males and females [P = 0.392; chi 2 test]. Helminth parasites of B. viridis were the monogenean, Polystoma viridis, the digenean, Haplometra cylindracea the cyclophyllid cestodes, Nematotaenia dispar; Diplopylidium acanthotetra cysticercoid and the nematodes, Cosmocerca ornata, Cosmocerca commutata, Rhabdias bufonis and Aplectana sp. There was no significant difference between the rate of infection with P. viridis [chi 2 = 1.92, P = 0.166] and N. dispar [chi 2 = 0.032, P = 0.859] in males and females. Males [n = 12], however, were significantly more infected [chi 2 = 5.52, P = 0.019] with R. bufonis than females [n = 8]. SVL of B. viridis infected with N. dispar and uninfected samples were significantly different [P = 0.016]. No significant difference was shown for infection with P. viridis [P>0.05; Student's t-test]. Among 17 marsh frogs [Rana ridibunda ridibunda], 58.8% were infected with helminth parasites. There was no significant difference between SVL of infected [n = 10] and uninfected [n = 7] frogs [P = 0.238; Student's t-test] and also between uninfected males and females using [P = 0.201; chi 2 test]. Metacercarian cysts of digenetic trematode Codonocephalus urniger and the nematode Aplectana sp. were found in Rana ridibunda ridibunda specimens. Twelve true tree frogs, Hiyla arborea savignyi were examined. The monogenean, Polystoma viridis, the cyclophyllid cestode, Nematotaenia dispar and the nematode, Aplectana sp. were found in the tree frogs
Subject(s)
Animals , Helminths , Bufonidae/parasitology , Ranidae/parasitology , Anura/classification , Anura/parasitologyABSTRACT
A large pleomorphic trypanosome [identified as Trypanosoma mega] was described from the toad Bufo viridis collected from Siwah Oasis at the Western Desert of Egypt. The prevalence of the trypanosome was 83.3%. Three trypanosome forms were described [small, intermediate and large stumpy form]. Observations were also made on the lysed [diffused] trypanosomes. This was the first record of T. mega from B. viridis in Egypt, which represents a new host and new geographical location. The measurements of the present trypanosome were given and compared with related forms previously described from Egypt