Rodrigo A. F. Redondo

Analysis of the first and second internal transcribed spacer sequences of the ribosomal DNA in Biomphalaria tenagophila complex (Mollusca: Planorbidae)

The first and second internal transcribed spacer regions (ITS1 and ITS2) of the ribosomal DNA of Biomphalaria tenagophila complex (B. tenagophila, B. occidentalis, and B. t. guaibensis) were sequenced and compared. The alignment lengths of these regions were about 655 bp and 481 bp, respectively. Phylogenetic relationships among the Biomphalaria species were inferred by Maximum Parsimony and Neighbor-joining methods. The phylogenetic trees produced, in most of the cases, were in accordance with morphological systematics and other molecular data previously obtained by polymerase chain reaction and restriction fragment length polymorphism analysis. The present results provide support for the proposal that B. tenagophila represents a complex comprising B. tenagophila, B. occidentalis and B. t. guaibensis.

Dominant culturable bacterial microbiota in the digestive tract of the American black vulture (Coragyps atratus Bechstein 1793) and search for antagonistic substances

Strict and facultative culturable anaerobic bacteria from the digestive tract of six American black vultures (Coragyps atratus Bechstein 1793) were isolated and identified. After capture, the birds received a non-contaminated diet for one week to eliminate possible allochthonous microorganisms. Then, specimens collected from tongue, stomach and intestines were weighed, submitted to decimal dilution in an anaerobic chamber, inoculated into culture media and incubated aerobically and anaerobically at 37oC for enumeration, isolation and identification. Isolated bacteria were submitted to tests to detect possible antagonisms between them. The total bacterial population along the digestive tract ranged from 3.46 ± 0.39 log CFU/g in the stomach to 10.75 ± 0.37 log CFU/g in the distal intestine. Some bacteria were isolated for the first time from the digestive tract of C. atratus: Actinomyces bovis, Lactobacillus cellobiosus, Micrococcus luteus, Neisseria sicca, Clostridium bifermentans, Enterobacter agglomerans, Peptostreptococcus sp., Sarcina sp., Serratia odorifera, and Shigella flexneri. Associations between microorganisms were observed during isolation on two occasions, one involving A. bovis and N. sicca, and the other involving A. bovis and a Gram-negative rod. Hetero-, iso- and autoantagonisms were observed, suggesting the ecological role of these indigenous microorganisms in terms of population auto-control and environmental barrier in the digestive tract of carrion-feeding birds.

Prospection, structural analysis and phylogenetic relationships of endogenous γ-phospholipase A2 inhibitors in Brazilian Bothrops snakes (Viperidae, Crotalinae)

During the last 20 years, there have been an increasing number of reports on endogenous phospholipase A(2) inhibitors (PLIs) in the sera of snakes. These studies have demonstrated the existence of three different structural classes of PLIs (alpha, beta and gamma). The gamma class members are potent inhibitors of phospholipases A(2) (PLA(2)) from the venom of Viperidae snakes. These enzymes, together with the mammalian pro-inflammatory PLA(2), belong to the IIA class of the PLA(2)-superfamily. Although coming from distinct sources, these phospholipases A(2) share main structural features. For this reason, gammaPLIs have been considered as potential models for the development of selective inhibitors of pro-inflammatory PLA(2) in humans. In spite of the rich diversity of the ophidian fauna in Brazil, only two gammaPLI representatives, from Crotalus durissus terrificus and Lachesis muta, have been described in Brazilian snakes so far. Here we investigated the presence of transcripts of novel gammaPLIs in six Bothrops species (Viperidae, Crotalinae) commonly found in our country: Bothrops alternatus, Bothrops erythromelas, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi. gammaPLI transcripts were present in every species analysed. The deduced mature proteins possessed 181 amino acid residues following a 19-residue signal peptide, similar to the gammaPLIs from C. d. terrificus taken as our model, with the exception of the deduced proteins from B. erythromelas and B. neuwiedi snakes. In these particular cases, an insertion of 4-amino acid residues was consistently present. A Bayesian tree was obtained for the Brazilian Bothrops gammaPLIs, showing four clusters: (1) L. muta and B. jararacussu, (2) B. alternatus, (3) B. erythromelas and B. neuwiedi, (4) B. jararaca and B. moojeni. Detailed structural analysis and further comparisons of these novel Bothrops inhibitors with gammaPLIs from New and Old World snakes are provided.

CYBB, an NADPH-oxidase gene: restricted diversity in humans and evidence for differential long-term purifying selection on transmembrane and cytosolic domains†‡

CYBB encodes the gp91‐phox protein of the phagocytic NADPH oxidase; the innate immunity‐related enzymatic complex responsible for the respiratory burst. Mutations in CYBB can cause chronic granulomatous disease (CGD), a primary immunodeficiency characterized by ineffective microbicidal activity, for which over 150 family‐specific mutations have been described. It is also plausible that common SNPs in CYBB alter the expression or function of gp91‐phox, determining differences in susceptibility to complex disorders such as autoimmune or infectious diseases. We have resequenced the exons, UTRs, and intronic regions of CYBB in 102 ethnically diverse individuals and genotyped nine tag‐SNPs in 942 individuals from 52 worldwide populations. The 28 observed SNPs (none of which nonsynonymous) reside on 28 haplotypes that can be collapsed into five clades. CYBB shows lower diversity than other X‐chromosome genes and most of the between‐population genetic variance was observed among Africans and non‐Africans. The African population shows the highest diversity and the lowest linkage disequilibrium (LD). Because there is extensive shared LD among non‐Africans, tag‐SNPs can be effectively employed in gene‐centric association studies and are portable across Eurasian and Native American populations. Comparison of CYBB coding sequences among mammals evidences the action of long‐term purifying selection, which is stronger on the C‐terminal cytosolic domain than on the N‐terminal transmembrane domain of gp91‐phox. Hum Mutat 29(5), 623–632, 2008. Published 2008 Wiley‐Liss, Inc.

Remaining genetic diversity in Brazilian Merganser (Mergus octosetaceus)

The Brazilian Merganser is a very rare and threatened species that nowadays inhabits only a few protected areas and their surroundings in the Brazilian territory. In order to estimate the remaining genetic diversity and population structure in this species, two mitochondrial genes were sequenced in 39 individuals belonging to two populations and in one individual collected in Argentina in 1950. We found a highly significant divergence between two major remaining populations of Mergus octosetaceus, which suggests a historical population structure in this species. Furthermore, two deeply divergent lineages were found in a single location, which could due to current or historical secondary contact. Based on the available genetic data, we point out future directions which would contribute to design strategies for conservation and management of this threatened species.