Sergio Echeverrigaray

Phylogeography of the Bothrops jararaca complex (Serpentes: Viperidae): past fragmentation and island colonization in the Brazilian Atlantic Forest

The Brazilian Atlantic Forest is one of the worlds major biodiversity hotspots and is threatened by a severe habitat loss. Yet little is known about the processes that originated its remarkable richness of endemic species. Here we present results of a large‐scale survey of the genetic variation at the mitochondrial cytochrome b gene of the pitviper, jararaca lancehead (Bothrops jararaca), and two closely related insular species (Bothrops insularis and Bothrops alcatraz), endemic of this region. Phylogenetic and network analyses revealed the existence of two well‐supported clades, exhibiting a southern and a northern distribution. The divergence time of these two phylogroups was estimated at 3.8 million years ago, in the Pliocene, a period of intense climatic changes and frequent fragmentation of the tropical rainforest. Our data also suggest that the two groups underwent a large size expansion between 50 000 and 100 000 years ago. However, the southern group showed a more marked signal of population size fluctuation than the northern group, corroborating evidences that southern forests may have suffered a more pronounced reduction in area in the late Pleistocene. The insular species B. alcatraz and B. insularis presented very low diversity, each one sharing haplotypes with mainland individuals placed in different subclades. Despite their marked morphological and behavioural uniqueness, these two insular species seem to have originated very recently and most likely from distinct costal B. jararaca populations, possibly associated with late Pleistocene or Holocene sea level fluctuations.

Micropropagation of Cunila galioides, a popular medicinal plant of south Brazil

Axillary buds of field plants of Cunila galioides Benth. were used to evaluate the effect of growth regulators and culture media on the in vitro shoot proliferation and growing. The highest multiplication rate was obtained using Murashige and Skoog (MS) medium supplemented with 8.8 μM of benzyladenine. Repeated subcultures of shoot tips and single nodes at 4-week intervals for eight months on the above medium enabled mass multiplication of shoots without any evidence of decline. The best conditions for rooting were MS medium plus 0.5 to 2.5 μM of indolebutyric acid. The rooted plants were successfully transferred to soil, exhibiting a normal development.

The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC)

Nodal segments from micropropagated plants were used to evaluate the effect of growth regulators on the in vitro shoot proliferation and rooting of Lavandula vera DC. The highest multiplication rate was obtained using MS medium supplemented with 1.0 mg l-1 of TDZ (2.25 μM) or BA (2 μM). Hyperhydricity occurred at high concentrations of these growth regulators. Rooting of the plantlets was obtained in all the media evaluated. However, rooting rates and root growth increased with increased concentrations of NAA and the reduction of the salt strength of the media. The plantlets were successfully transferred to soil and grown to maturity, exhibiting a normal development, with high uniformity and no evidences of somaclonal variation.

Nematicidal activity of monoterpenoids against the root-knot nematode Meloidogyne incognita.

ABSTRACT Nematicidal activity of 22 monoterpenoids were evaluated in vitro and in pot experiments. Twenty of the twenty-two monoterpenoids significantly reduced hatching, and 11 reduced J2 mobility of the root-knot nematode Meloidogyne incognita at a concentration of 250 mg/liter. In general, compounds with hydroxyl and carbonyl groups exhibited higher nematicidal activity than other terpenoids. Borneol, carveol, citral, geraniol, and alpha-terpineol showed the highest nematicidal activity among the in vitro tested monoterpenoids. These compounds exhibited a dose dependent effect, and drastically reduced eggs hatching and J2 viability at low concentrations. These monoterpenoids, at 100 and 250 mg/kg concentration, diminished root galling of tomato plants in pot experiments. The results suggest that the selected monoterpenoids, and essential oils with high concentration of these compounds, are potential nematicides against Meloidogyne.

Genetic diversity among Brazilian cultivars and landraces of tomato Lycopersicon esculentum Mill. revealed by RAPD markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.

Aeromonas associated diarrhoeal disease in south Brazil: prevalence, virulence factors and antimicrobial resistance

Aeromonas were isolated from 27 (6.6%) of 408 patients admitted with acute gastroenteritis in two hospitals at Rio Grande do Sul, Brazil. Isolates were classified as A. hydrophila (51.8%), A. caviae (40.8%), and A. veronii biotype sobria (7.4%). The highest prevalence of Aeromonas associated infections occurred in lactants and children. Virulence genes (aerA -aerolysin/hemolysin, ahpA -serine-protease, satA - glycerophospholipid-cholesterol acyltransferase, lipA -lipase, and ahyB -elastase) and virulence factors (hemolytic, proteolitic, lipolitic activities, and biofilm formation) were identified in most A. hydrophila and A. veronii biotype sobria isolates, with lower frequencies on A. caviae. All Aeromonas isolates were resistant to ampicillin, ticarcillin/clavulanic acid, cephalotin, and cephazolin, and most of them (>70%) exhibited resistance to imipenem, carbenicillin, amoxillin/sulbactan, and piperacillin. Multiple-resistance, more than four antibiotics, was evidenced in 29.6% of the isolates. The most efficient antibiotics were the quinolones (ciprofloxacin and norfloxacin), and the aminoglycosides (amikacin and netilmicin).

Pathogenic factors and antimicrobial resistance of Staphylococcus epidermidis associated with nosocomial infections occurring in intensive care units

Nosocomial infections constitute an important problem in hospitals, intensive care units (ICU) having the highest incidence of this type of infection. Staphylococci, especially Staphylococcus epidermidis and Staphylococcus aureus, are among the most important microorganisms associated with nosocomial infections. S. epidermidis is a common skin resident, and can be introduced into the clinical environment by patients and hospital staff. The situation in hospitals is aggravated by the emergence of multiresistant strains. We evaluated 98 hospital S. epidermidis isolates collected at neonatal, pediatric and adult ICUs and 20 S. epidermidis control skin resident isolates from healthy volunteers, for resistance to ten antibiotics and chemotherapeutic agents, and other pathogenicity factors. A high frequency (76.5%) of multiresistance was detected in clinical isolates, whereas community isolates were resistant to penicillin and ampicillin only. The frequency of multiresistant strains was 67.7% in the neonatal ICU, 66.6% in the pediatric ICU and 60.8% in the adult ICU, the lower frequency of multiresistant isolates in the adult ICU indicates a higher incidence of community strains in this unit. There were significantly higher frequencies of hemolytic, proteolytic and biofilm-forming isolates in the clinical isolates than the community isolates, indicating a higher incidence of strains with pathogenic potential in the hospital environment. Except for slight correlation with hemolytic activity there was no correlation between antibiotic multiresistance and pathogenicity factors.

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccards coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.

Random amplified polymorphic DNA (RAPD) markers variability among cultivars and landraces of common beans (Phaseolus vulgaris L.) of south-Brazil

To evaluate the variability among cultivars and landraces of common bean(Phaseolus vulgaris L.), 15 cultivars and 18 landraces of common bean (Phaseolus vulgarisL.), a undefined species of Phaseolus,two landraces of Vigna angularis L., and a landrace of soybean (Glycine maxL.), were screened with fifteen oligonucleotide primers in PCR reactions. An average of 20.3 RAPD bands were scored per primer. A total of 304 amplification products were scored of which 88.8% were polymorphic among Phaseolus genotypes. Based on the RAPD markers, four major clusters were formed. Three clusters corresponded to the soybean, to the two Vigna angularis landraces, and to the Phaseolus sp. landrace, respectively. The fourth cluster include all the landraces and cultivars of Phaseolus vulgaris. This large group could be separated into three subgroups that were correlated with the phaseolin patterns and the average seed weight of the genotypes. The analysis shows that most of the landraces collected in South Brazil (17 out of 18) belong to the Andean gene pool, and most of the cultivars (13 out of 15) belong to the Middle American gene pool.

Essential oil composition of south Brazilian populations of Cunila galioides and its relation with the geographic distribution

Abstract Air-dried samples of 20 Brazilian populations of Cunila galioides were extracted by steam distillation and analyzed using GS and GS–MS. A total of 40 volatile compounds were detected and identified. Fourteen compounds were presented in high concentration, (>10% of the essential oils) in at least one of the populations, and were used to analyze the relationship between the populations. Three affinity groups, which can be considered as chemotypes, were formed by the average linkage cluster analysis. The citral group was characterized by high concentrations of neral and geranial (28 ± 1.49% and 40.5 ± 1.73%, respectively), the ocimene group by high concentration of trans-β-ocimene (33.05 ± 2.54%), and the menthene group by the presence of 1,8-cineole (10.69 ± 2.93%), trans-p-2,8-menthadiene-1-ol (11.46 ± 1.54%), 1,3,8-menthatriene (10.39 ± 1.41%), and 1,5,8-p-menthatriene (7.75 ± 2.32%), as their main constituents. Geographically, the chemotypes containing citral originated from the northeast plateau of Rio Grande do Sul, whereas chemotypes containing ocimene originated from the grasslands of high altitudes of the Atlantic range, and those of the menthene group were found in a transition area between the two regions. The chemovariation observed appear to be genetically determined.