Janaina da Costa de Noronha

Antiproliferative activity of Rhinella marina and Rhaebo guttatus venom extracts from Southern Amazon

The venom of amphibians is a fascinating source of active substances. In view of their medical importance and aiming to explore the amazing Brazilian biodiversity, we conducted bioprospecting of antiproliferative activity in extracts of Rhinella marina and Rhaebo guttatus toads occurring in the Southern Amazon of Mato Grosso, Brazil. LC-MS and HPLC analysis of the venom extracts of R. marina revealed four bufadienolides (telocinobufagin, marinobufagin, bufalin and resibufogenin. R. guttatus venom extracts contained only marinobufagin. First, R. marina and R. guttatus venom extracts were evaluated for cytotoxicity against tumor cell lines by the MTT assay. All extracts revealed cytotoxicity, where R. marina extracts were comparable to doxorubicin (IC₅₀ values ranging from 0.01 to 0.23 μg/mL). Only extracts of R. guttatus toad venom caused membrane disruption of human erythrocytes. The extracts were investigated for selective activity by determining their effect on stimulated human peripheral blood mononuclear cells (PBMC) with the Alamar Blue™ assay. The extracts were up to 80-fold more selective against leukemia cells when compared to dividing leukocytes. Aiming to confirm these antiproliferative effects, BrdU incorporation into DNA was measured in HL-60 treated cells with R. marina venom extracts. These extracts decreased BrdU incorporation at both concentrations tested. In summary, nine extracts of R. marina and R. guttatus venom showed pronounced lethal and discriminating effects on tumor lines, especially those from R. marina, highlighting toad parotoid gland secretions as a promising source for novel lead anticancer chemicals.

Floral resource partitioning by ants and bees in a jambolan Syzygium jambolanum (Myrtaceae) agroforestry system in Brazilian Meridional Amazon

The fruit production of flowering plants critically depends on the pollination services provided by animals that compete for flower resources. The output of competitive interactions between ants and bees for inflorescences of jambolan Syzygium jambolanum (Myrtaceae) in an agroforestry system in Brazilian Meridional Amazonian are an interesting system of investigation due the possibility to control variables experimentally. In 20 S. jambolanum individuals we performed 300 treatments in different inflorescences of two strata (upper and lower) as follows: (1) ants exclusion, (2) bees exclusion, and (3) control group where ants and bees could access the inflorescences. There was no difference in the number of inflorescences, volume of nectar and sugar concentration between the strata. Also the visitors considered are distributed equally in the tree’s stratum. When bees were prevented from access the inflorescences, ants dominated more inflorescences only in the lower stratum. On the contrary, when ants were excluded, bees visited more inflorescences only in the upper stratum. We conclude that ants prevent the access to bees and vice versa as the result of different ability of resource utilization and foraging strategies. Thus, preventing the access of ants to the floral nectar could increase the level of nectar available to pollinators of S. jambolanum, thereby increasing productivity and reducing economic losses.

Amphibia, Anura, Microhylidae, Chiasmocleis avilapiresae Peloso and Sturaro, 2008 and Chiasmocleis bassleri Dunn, 1949: New Distribution records

This note reports the presence of Chiasmocleis bassleeri and C. avilapiresae in the north of Mato Grosso state. It extends the distribution of C. avilapiresae and C. bassleri 140 km from the nearest known locality (Aripuana, MT, Brazil) and 1170 and 1350 km from the type-locality (Floresta Nacional de Caxiuana, state of Para, Brazil and Rio Utoquinia to Rio Tapiche, Peru, respectively).

Predation of bat (Molossus molossus: Molossidae) by the centipede Scolopendra viridicornis (Scolopendridae) in Southern Amazonia

Centipedes are opportunistic carnivore predators, and large species can feed on a wide variety of vertebrates, including bats. The aim of this study was to report the third record of bat predation by centipedes worldwide, the first record in the Amazon region, while covering aspects of foraging, capture and handling of prey. We observed the occurence in a fortuitous encounter at Cristalino State Park, located in the Amazon region of the state of Mato Grosso, Brazil. The attack took place in a small wooden structure, at about three meters from the floor, and was observed for 20 minutes. During the observation, the centipede stung the neck and abdominal region of the bat several times, grabbing the prey with its 15 pairs of front legs while hanging from the ceiling with its hind legs. This type of observation suggests that vertebrates can be important preys for invertebrates such as giant centipedes, both for its nutritional composition and for the amount of energy available in a single prey.

Amblyomma tick species infesting amphibians and reptiles in the seasonally dry Amazon forest, with new host records for Amblyomma rotundatum (Acari: Ixodida: Ixodidae)

Abstract Our study yielded a list of ticks found on amphibians and reptiles at five sites within seasonally dry Amazon forest in Mato Grosso State, central Brazil, in addition to new host records for A. rotundatum. We collected 431 tick specimens: 79 larvae, 115 nymphs, 38 females, and 199 males belonging to A. humerale, A. rotundatum, and Amblyomma sp., on 39 anurans and 21 reptiles. The toads R. guttatus and R. margaritifera and the frog L. pentadactylus are new hosts for A. rotundatum.

Antiplasmodial and Cytotoxic Activities of Toad Venoms from Southern Amazon, Brazil

The drug-resistance of malaria parasites is the main problem in the disease control. The huge Brazilian biodiversity promotes the search for new compounds, where the animal kingdom is proving to be a promising source of bioactive compounds. The main objective of this study was to evaluate the antiplasmodial and cytotoxic activity of the compounds obtained from the toad venoms of Brazilian Amazon. Toad venoms were collected from the secretion of Rhinella marina and Rhaebo guttatus in Mato Grosso State, Brazil. The powder was extracted at room temperature, yielding 2 extracts (RG and RM) and a substance (‘1’) identified as a bufadienolide, named telocinobufagin. Growth inhibition, intraerythrocytic development, and parasite morphology were evaluated in culture by microscopic observations of Giemsa-stained thin blood films. Cytotoxicity was determined against HepG2 and BGM cells by MTT and neutral red assays. The 2 extracts and the pure substance (‘1’) tested were active against chloroquine-resistant Plasmodium falciparum strain, demonstrating lower IC50 values. In cytotoxic tests, the 2 extracts and substance ‘1’ showed pronounced lethal effects on chloroquine-resistant P. faciparum strain and low cytotoxic effect, highlighting toad parotoid gland secretions as a promising source of novel lead antiplasmodial compounds.

Amphibia, Anura, Hylidae, Phyllomedusa boliviana Boulenger, 1902 and Phyllomedusa camba De la Riva, 2000: distribution extension in central Brazil

This note reports the presence of Phyllomedusa camba in the municipality of Cotriguacu (Amazon forest) and P. boliviana in the municipality of Poxoreu (Cerrado), state of Mato Grosso, central Brazil. This represents the first record for Phyllomedusa boliviana and a distribution extension of P. camba in the state Mato Grosso.

Reproductive behaviour of the glass frog Hyalinobatrachium cappellei (Anura: Centrolenidae) in the Southern Amazon

ABSTRACT Frog species of the Centrolenidae family exhibit a high variation in reproductive behaviours. Herein we describe the reproductive, ecological, acoustic and behavioural features of Hyalinobatrachium cappellei in Southern Amazon. Two hypotheses were also evaluated: (1) are ecological characteristics of the reproductive site related to male size and quantity of eggs and clutches? And (2) do males with one or more existing clutches at reproductive sites accumulate more new clutches than those males initially without them? This study was conducted at two streams, with sampling at 1 km each in Southern Amazon. Species reproduction occurred during the wet season and was considered prolonged breeding. Although male vocalization occurred on both the upper and underside of leaves, clutches were most often deposited on the underside of leaves. Males emitted a courtship call upon female approach. The process of female approach until oviposition lasted approximately eight hours. After oviposition, females left the breeding site while males remained near the clutch, covering it with the belly in ventral brooding behaviour. Thus, egg attendance is reported for the first time for this species. No intraspecific or interspecific aggressive behaviour was recorded. There was no relationship between habitat characteristics of the reproductive sites and male size, number of eggs and clutches. However, after 10 days of monitoring, we found evidences that parental males had a higher number of clutches than those males initially without clutches. Similar to other glass frog species, the behavioural characteristics of H. cappellei make this species an excellent model to use in evaluating the cost-effectiveness of parental care and the importance of egg attendance for offspring survival. Therefore, these data contribute towards a better understanding of the complex phylogenetic and biogeographic relationships between centrolenid lineages.

Marinobufagin, a molecule from poisonous frogs, causes biochemical, morphological and cell cycle changes in human neoplasms and vegetal cells

Skin toad secretion present physiologically active molecules to protect them against microorganisms, predators and infections. This work detailed the antiproliferative action of marinobufagin on tumor and normal lines, investigate its mechanism on HL-60 leukemia cells and its toxic effects on Allium cepa meristematic cells. Initially, cytotoxic action was assessed by colorimetric assays. Next, HL-60 cells were analyzed by morphological and flow cytometry techniques and growing A. cepa roots were examined after 72 h exposure. Marinobufagin presented high antiproliferative action against all human tumor lines [IC50 values ranging from 0.15 (leukemia) to 7.35 (larynx) μM] and it failed against human erythrocytes and murine lines. Human normal peripheral blood mononuclear cells (PBMC) were up to 72.5-fold less sensitive [IC50: 10.88 μM] to marinobufagin than HL-60 line, but DNA strand breaks were no detected. Leukemia treaded cells exhibited cell viability reduction, DNA fragmentation, phosphatidylserine externalization, binucleation, nuclear condensation and cytoplasmic vacuoles. Marinobufagin also reduced the growth of A. cepa roots (EC50: 7.5 μM) and mitotic index, caused cell cycle arrest and chromosomal alterations (micronuclei, delays and C-metaphases) in meristematic cells. So, to find out partially targeted natural molecules on human leukemia cells, like marinobufagin, is an amazing and stimulating way to continue the battle against cancer.