Identification and lineage genotyping of South American trypanosomes using fluorescent fragment length barcoding

Trypanosoma cruzi and Trypanosoma rangeli are human-infective blood parasites, largely restricted to Central and South America. They also infect a wide range of wild and domestic mammals and are transmitted by a numerous species of triatomine bugs. There are significant overlaps in the host and geographical ranges of both species. The two species consist of a number of distinct phylogenetic lineages. A range of PCR-based techniques have been developed to differentiate between these species and to assign their isolates into lineages. However, the existence of at least six and five lineages within T. cruzi and T. rangeli, respectively, makes identification of the full range of isolates difficult and time consuming. Here we have applied fluorescent fragment length barcoding (FFLB) to the problem of identifying and genotyping T. cruzi, T. rangeli and other South American trypanosomes. This technique discriminates species on the basis of length polymorphism of regions of the rDNA locus. FFLB was able to differentiate many trypanosome species known from South American mammals: T. cruzi cruzi. T. cruzi marinkellei, T. dionisii-like, T. evansi, T. lewisi, T. rangeli, T. theileri and T. vivax. Furthermore, all five T. rangeli lineages and many T. cruzi lineages could be identified, except the hybrid lineages TcV and TcVI that could not be distinguished from lineages III and II respectively. This method also allowed identification of mixed infections of T. cruzi and T. rangeli lineages in naturally infected triatomine bugs. The ability of FFLB to genotype multiple lineages of T. cruzi and T. rangeli together with other trypanosome species, using the same primer sets is an advantage over other currently available techniques. Overall, these results demonstrate that FFLB is a useful method for species diagnosis, genotyping and understanding the epidemiology of American trypanosomes. (C) 2010 Elsevier B.V. All rights reserved.

Phylogenetic analysis of Trypanosoma vivax supports the separation of South American/West African from East African isolates and a new T-vivax-like genotype infecting a nyala antelope from Mozambique

In this study, we addressed the phylogenetic and taxonomic relationships of Trypanosoma vivax and related trypanosomes nested in the subgenus Duttonella through combined morphological and phylogeographical analyses. We previously demonstrated that the clade T. vivax harbours a homogeneous clade comprising West African/South American isolates and the heterogeneous East African isolates. Herein we characterized a trypanosome isolated from a nyala antelope (Tragelaphus angasi) wild-caught in Mozambique (East Africa) and diagnosed as T. vivax-like based on biological, morphological and molecular data. Phylogenetic relationships, phylogeographical patterns and estimates of genetic divergence were based on SSU and ITS rDNA sequences of T. vivax from Brazil and Venezuela (South America), Nigeria (West Africa), and from T. vivax-like trypanosomes from Mozambique, Kenya and Tanzania (East Africa). Despite being well-supported within the T. vivax clade, the nyala trypanosome was highly divergent from all other T. vivax and T. vivax-like trypanosomes, even those from East Africa. Considering its host origin, morphological features, behaviour in experimentally infected goats, phylogenetic placement, and genetic divergence this isolate represents a new genotype of trypanosome closely phylogenetically related to T. vivax. This study corroborated the high complexity and the existence of distinct genotypes yet undescribed within the subgenus Duttonella.

Evolutionary history of trypanosomes from South American caiman (Caiman yacare) and African crocodiles inferred by phylogenetic analyses using SSU rDNA and gGAPDH genes

In this study, using a combined data set of SSU rDNA and gGAPDH gene sequences, we provide phylogenetic evidence that supports Clustering of crocodilian trypanosomes from the Brazilian Caiman yacare (Alligatoridae) and Trypanosoma grayi, a species that Circulates between African crocodiles (Crocodilydae) and tsetse flies. In a survey of trypanosomes in Caiman yacare from the Brazilian Pantanal, the prevalence of trypanosome infection was 35% as determined by microhaematocrit and haemoculture, and 9 cultures were obtained. The morphology of trypomastigotes from caiman blood and tissue imprints was compared with those described for other crocodilian trypanosomes. Differences in morphology and growth behaviour of caiman trypanosomes were corroborated by molecular polymorphism that revealed 2 genotypes. Eight isolates were ascribed to genotype Cay01 and 1 to genotype Cay02. Phylogenetic inferences based on concatenated SSU rDNA and gGAPDII sequences showed that caiman isolates are closely related to T. grayi, constituting a well-supported monophyletic assemblage (clade T. grayi). Divergence time estimates based on clade composition, and biogeographical and geological events were used to discuss the relationships between the evolutionary histories of crocodilian trypanosomes and their hosts.

The South American Plasmodium falciparum var gene repertoire is limited, highly shared and possibly lacks several antigenic types

The Plasmodium falciparum var gene family encodes large variant antigens, which are important virulence factors, and also targets of the humoral host response. The frequently observed mild outcomes of falciparum malaria in many places of the Amazon area prompted us to ask whether a globally restricted variant (var) gene repertoire is present in currently circulating and older isolates of this area. By exhaustive analysis of var gene tags from 89 isolates and clones taken during many years from all over the Brazilian Amazon, we estimate that there are probably no more than 350-430 distinct sequence types, less than for any similar sized area studied so far. Detailed analysis of the var tags from genetically distinct clones obtained from single isolates revealed restricted and redundant repertoires suggesting either a low incidence of infective bites or restricted variant gene diversity in inoculated parasites. Additionally, we found a structuring of var gene repertoires observed as a higher pairwise typing sharing in isolates from the same microregion compared to isolates from different regions. Fine analysis of translated var tags revealed that certain Distinct Sequence Identifiers (DSIDs) were differently represented in Brazilian/South American isolates when compared to datasets from other continents. By global alignment of worldwide var DBL alpha sequences and sorting in groups with more than 76% identity, 125 clusters were formed and more than half of all genes were found in nine clusters with 50 or more sequences. While Brazilian/South American sequences were represented only in 64 groups, African sequences were found in the majority of clusters. DSID type 1 related sequences accumulated almost completely in one single cluster, indicating that limited recombination occurs in these specific var gene types. These data demonstrate the so far highest pairwise type sharing values for the var gene family in isolates from all over an entire subcontinent. The apparent lack of specific sequences types suggests that the P. falciparum transmission dynamics in the whole Amazon are probably different from any other endemic region studied and possibly interfere with the parasite`s ability to efficiently diversify its variant gene repertoires. (C) 2010 Elsevier B.V. All rights reserved.

The basement of the Punta del Este Terrane (Uruguay): an African Mesoproterozoic fragment at the eastern border of the South American Rio de La Plata craton

The Punta del Este Terrane (eastern Uruguay) lies in a complex Neoproterozoic (Brasiliano/Pan-African) orogenic zone considered to contain a suture between South American terranes to the west of Major Gercino-Sierra Ballena Suture Zone and eastern African affinities terranes. Zircon cores from Punta del Este Terrane basement orthogneisses have U-Pb ages of ca. 1,000 Ma, which indicate an lineage with the Namaqua Belt in Southwestern Africa. U-Pb zircon ages also provide the following information on the Punta del Este terrane: the orthogneisses containing the ca. 1,000 Ma inheritance formed at ca. 750 Ma; in contrast to the related terranes now in Africa, reworking of the Punta del Este Terrane during Brasiliano/Pan-African orogenesis was very intense, reaching granulite facies at ca. 640 Ma. The termination of the Brasiliano/Pan-African orogeny is marked by formation of acid volcanic and volcanoclastic rocks at ca. 570 Ma (Sierra de Aguirre Formation), formation of late sedimentary basins (San Carlos Formation) and then intrusion at ca. 535 Ma of post-tectonic granitoids (Santa Teresa and Jos, Ignacio batholiths). The Punta del Este Terrane and unrelated western terranes represented by the Dom Feliciano Belt and the Rio de La Plata Craton were in their present positions by ca. 535 Ma.

Orbitally driven east-west antiphasing of South American precipitation

The variations of tropical precipitation are antiphased between the hemispheres on orbital timescales. This antiphasing arises through the alternating strength of incoming solar radiation in the two hemispheres, which affects monsoon intensity and hence the position of the meridional atmospheric circulation of the Hadley cells(1-4). Here we compare an oxygen isotopic record recovered from a speleothem from northeast Brazil for the past 26,000 years with existing reconstructions of precipitation in tropical South America(5-8). During the Holocene, we identify a similar, but zonally oriented, antiphasing of precipitation within the same hemisphere: northeast Brazil experiences humid conditions during low summer insolation and aridity when summer insolation is high, whereas the rest of southern tropical South America shows opposite characteristics. Simulations with a general circulation model that incorporates isotopic variations support this pattern as well as the link to insolation-driven monsoon activity. Our results suggest that convective heating over tropical South America and associated adjustments in large-scale subsidence over northeast Brazil lead to a remote forcing of the South American monsoon, which determines most of the precipitation changes in the region on orbital timescales.

Reproductive Patterns and Birth Seasonality in a South-American Breeding Colony of Common Marmosets, Callithrix jacchus

SOUSA,M.B.C. et al. Reproductive Patterns and Birth Seasonality in a South-American Breeding Colony of Common Marmosets, Callithrix jacchus. Primates, v.40, n.2, p. 327-336, Apr. 1999.

Free- and peptide-based dietary arginine supplementation for the South American fish pacu (Piaractus mesopotamicus)

Arginine was hypothesized to be a model compound in the present study on molecular forms of indispensable amino acid (IAA) dietary supplementation. Juvenile South American pacu (Piaractus mesopotamicus) were fed diets containing arginine in a protein base (casein-wheat gluten or casein-gelatin), or the casein-wheat gluten base supplemented with dipeptide or free arginine at two levels (5 and 10 g kg(-1)). Growth and protein efficiency ratios were significantly affected by diets, but not by arginine molecular form. Three free dispensable amino acids (DAA) and four IAA in plasma were affected by diet, but plasma arginine concentrations did not differ. Plasma urea concentrations, being very low in the pacu, and hepatic arginase activities, were not affected by diet (P = 0.10-0.11), but together with plasma ornithine, mirrored the growth data. Molecular form of arginine supplementation, free or dipeptide, significantly changed several free IAA (Phe, Leu, Ile, His) and urea, with a higher mean plasma concentration in dipeptide fed fish. The dietary treatments, or molecular form of the arginine supplementation, did not change proximate composition, except that calcium levels decreased with higher dietary arginine supplementation level. The present study indicates that dipeptides can provide IAA to pacu, and that arginine supplemented in this form is utilized as efficiently as in free form.

A comparative study of population traits between two South American populations of the striped-legged hermit crab Clibanarius vittatus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Growth, fat content and fatty acid profile of South American catfish, surubim (Pseudoplatystoma fasciatum) juveniles fed live, commercial and formulated diets

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of different dietary lipid sources on the survival, growth, and fatty acid composition of South American catfish, Pseudoplatystoma fasciatum, Surubim, juveniles

The present study examines the effect of four semi-purified diets (casein-gelatin based) where the source of fatty acids was free (esterified) oleic acid and linoleic acid (LA) (LOA diet), linseed and olive oil (predominantly LA and linolenic acid) (LO diet), cod liver oil (rich in highly unsaturated fatty acids) (CLO diet), and soybean lecithin (phospholipids; mostly LA) (LE diet) on the growth of juvenile South American catfish (surubim, Pseudoplatystoma fasciatum, Pimelodidae) (0.98 +/- 0.04 g individual weight). Fish were fed at a restricted-readjusted feeding rate for 8 wk. At the end of the experiment, LE-diet-fed fish grew significantly larger than those of the other three groups (P < 0.05). Considerable cannibalism was observed in all the treatments. It is suggested that the quantitative growth performance may possibly change under other conditions, with less or no cannibalism. Survival did not differ significantly among the fish fed four different diets. Muscle and liver lipid contents did not vary among dietary treatments (P > 0.05), but whole-body lipid concentrations were affected by dietary treatments. Fish fed LE diet contained significantly lower lipid level than those fed three other diets (P < 0.05). Muscle and liver fatty acid profiles reflected dietary fatty acid composition. Arachidonic acid level was significantly higher in muscle and liver of fish fed LOA and LE diets than in those fed LO and CLO diets. The results suggest that the efficiency of elongation and desaturation of 18C fatty acids depends on the dietary lipid source, and South American catfish has considerable capacity to transform linoleate to arachidonate.

Biology and fertility life table of the South American fruit fly Anastrepha fraterculus on grape

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

African-Derived mitochondria in South American native cattle breeds (Bos taurus): Evidence of a new taurine mitochondrial lineage

This article reports the nucleotide diversity within the control region of 42 mitochondrial chromosomes belonging to five South American native cattle breeds (Bos taurus). Analysis of these data in conjunction with B. taurus and B. indicus sequences from Africa, Europe, the Near East, India, and Japan allowed the recognition of eight new mitochondrial haplotypes and their relative positions in a phylogenetic network. The structure of genetic variation among different hypothetical groupings was tested through the molecular variance decomposition, which was best explained by haplotype group components. Haplotypes surveyed were classified as European-related and African-related. Unexpectedly, two haplotypes within the African cluster were more divergent from the African consensus than the latter from the European consensus. A neighbor-joining tree shows the position of two haplotypes compared to European/African mitochondrial lineage splitting. This different and putatively ancestral mitochondrial lineage (AA) is supported by the calibration of sequence divergence based on the Bos-Bison separation. The European/African mitochondria divergence might be subsequent (67,100 years before present) to that between AA and Africans (84,700 years before present), also preceding domestication times. These genetic data could reflect the haplotype distribution of Iberian cattle five centuries ago.

Restriction fragment length polymorphism (RFLP) in Exon 2 of the BoLA-DRB3 gene in South American cattle

The Bola-DRB3 gene participates in the development of the immune response and is highly polymorphic. For these reasons, it has been a candidate gene in studies of the genetic basis of disease resistance and in population genetic analysis. South American native cattle breeds have been widely replaced by improved exotic breeds leading to a loss of genetic resources. In particular South American native breeds have high levels of fertility and disease resistance. This work describes genetic variability in the BoLA-DRB3 gene in native (Caracu, Pantaneiro, Argentinean Creole) and exotic (Holstein, Jersey, Nelore, Gir) cattle breeds in Brazil and Argentina. PCR-RFLP alleles were identified by combining the restriction patterns for the BoLA-DRB3.2 locus obtained with RsaI, BstY, and HaeIII restriction enzymes. Allelic frequencies and deviations from the Hardy-Weinberg equilibrium were also calculated. Analysis of the 24 BoLA-DRB3 PCR-RFLP alleles identified showed differences in the allele distributions among breeds.

Gastro-intestinal transit time in South American deer

The study of diet and physiological peculiarities of the digestive system of neotropical deer is not well known and the literature shows inconsistencies. To better understand the digestive system of these mammals the difference in the gastro-intestinal transit time of four species of neotropical deer (Mazama americana, Mazama gouazoubira, Mazama nana, Blastocerus dichotomus), kept ill captivity, was evaluated. Four plants (Neonotonia wightii, Morus albans, Medicago sativa, Leucaena leucocephala) were utilized and two variables were measured, mean time of the beginning of the elimination (BE) and mean time of permanence (TP). The results obtained for BE indicated similarity among the deer species, with significant differences between M. gouazoubira (mean = 13.62 hr) and M. Americana (mean = 19.25 hr). For the plants, the BE was faster with N. wightii and L. leucocephala, and slower for M. sativa. The TP results for B. dichotomus showed longer time when compared to the other species, whereas M. gouazoubira had a lower permanence. Overall, N. wightii had the highest retention time in the digestive tract of all the deer species studied. Associated with this observation, N. wightii had the highest quantity of plant fiber of the plants tested. In a similar fashion M. sativa showed the lowest TP in the digestive tract of the deer and had the lowest quantity of acid detergent fiber. The data from this study showed that, within species, the shape of the excretion curve of the plants was similar when the animals consumed N. wightii or M. sativa. Blastocerus dichotomus and M. gouazoubira had the highest and the lowest gastro-intestinal transit time, respectively. This suggests that these species characterize different abilities to digest high fiber food, and consequently, represent the two extremities in the morphophysiological adaptation within the deer species evaluated. This information is vital because it is important to know the digestive physiology to define the diet of captive animals, particularly regarding the quantity and quality of fiber. Zoo Biol 25:47-57, 2006. (C) 2005 Wiley-Liss, Inc.