Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20%. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24%) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

Fitness evaluation of two Brazilian Aedes aegypti field populations with distinct levels of resistance to the organophosphate temephos

In Brazil, decades of dengue vector control using organophosphates and pyrethroids have led to dissemination of resistance. Although these insecticides have been employed for decades against Aedes aegypti in the country, knowledge of the impact of temephos resistance on vector viability is limited. We evaluated several fitness parameters in two Brazilian Ae. aegypti populations, both classified as deltamethrin resistant but with distinct resistant ratios (RR) for temephos. The insecticide-susceptible Rockefeller strain was used as an experimental control. The population presenting the higher temephos resistance level, Aparecida de Goiânia, state of Goiás (RR95 of 19.2), exhibited deficiency in the following four parameters: blood meal acceptance, amount of ingested blood, number of eggs and frequency of inseminated females. Mosquitoes from Boa Vista, state of Roraima, the population with lower temephos resistance level (RR95 of 7.4), presented impairment in only two parameters, blood meal acceptance and frequency of inseminated females. These results indicate that the overall fitness handicap was proportional to temephos resistance levels. However, it is unlikely that these disabilities can be attributed solely to temephos resistance, since both populations are also resistant to deltamethrin and harbour the kdr allele, which indicates resistance to pyrethroids. The effects of reduced fitness in resistant populations are discussed.

Susceptibility and resistance to deltamethrin of wild and domestic populations of Triatoma infestans (Reduviidae: Triatominae) in Bolivia: new discoveries

Bolivia is a high-endemic country for Chagas disease, for which the principal vector is Triatoma infestans (Triatominae). This is a mainly domestic species that is also found in the wild environment. Recently, an increasing number of studies have shown the importance of Triatominae resistance to insecticides, especially in Bolivia. Data regarding the susceptibility/resistance of wild and domestic populations of T. infestans to deltamethrin are presented. For the first time, domestic populations of the department of Santa Cruz were tested, showing low resistance. Although most of the wild populations were found to be susceptible to deltamethrin, three populations from three departments showed a mortality rate of less than 100%. This result is emphasised here.

Monitoring resistance to Bacillus thuringiensis subsp. israelensis in the field by performing bioassays with each Cry toxin separately

Bacillus thuringiensis subsp. israelensis (Bti) is increasingly used worldwide for mosquito control and is the only larvicide used in the French Rhône-Alpes region since decades. The artificial selection of mosquitoes with field-persistent Bti collected in breeding sites from this region led to a moderate level of resistance to Bti, but to relatively high levels of resistance to individual Bti Cry toxins. Based on this observation, we developed a bioassay procedure using each Bti Cry toxin separately to detect cryptic Bti-resistance evolving in field mosquito populations. Although no resistance to Bti was detected in none of the three mosquito species tested (Aedes rusticus, Aedes sticticus and Aedes vexans), an increased tolerance to Cry4Aa (3.5-fold) and Cry11Aa toxins (8-fold) was found in one Ae. sticticus population compared to other populations of the same species, suggesting that resistance to Bti may be arising in this population. This study confirms previous works showing a lack of Bti resistance in field mosquito populations treated for decades with this bioinsecticide. It also provides a first panorama of their susceptibility status to individual Bti Cry toxins. In combination with bioassays with Bti, bioassays with separate Cry toxins allow a more sensitive monitoring of Bti-resistance in the field.

Emerging Plasmodium vivax resistance to chloroquine in South America: an overview

The global emergence of Plasmodium vivax strains resistant to chloroquine (CQ) since the late 1980s is complicating the current international efforts for malaria control and elimination. Furthermore, CQ-resistant vivax malaria has already reached an alarming prevalence in Indonesia, East Timor and Papua New Guinea. More recently, in vivo studies have documented CQ-resistant P. vivax infections in Guyana, Peru and Brazil. Here, we summarise the available data on CQ resistance across P. vivax-endemic areas of Latin America by combining published in vivo and in vitro studies. We also review the current knowledge regarding the molecular mechanisms of CQ resistance in P. vivax and the prospects for developing and standardising reliable molecular markers of drug resistance. Finally, we discuss how the Worldwide Antimalarial Resistance Network, an international collaborative effort involving malaria experts from all continents, might contribute to the current regional efforts to map CQ-resistant vivax malaria in South America.

A novel ABCG-like transporter of Trypanosoma cruziis involved in natural resistance to benznidazole

Benznidazole (BZ) is one of the two drugs used for Chagas disease treatment. Nevertheless therapeutic failures of BZ have been reported, which were mostly attributed to variable drug susceptibility among Trypanosoma cruzistrains. ATP-binding cassette (ABC) transporters are involved in a variety of translocation processes and some members have been implicated in drug resistance. Here we report the characterisation of the first T. cruzi ABCG transporter gene, named TcABCG1, which is over-expressed in parasite strains naturally resistant to BZ. Comparison ofTcABCG1 gene sequence of two TcI BZ-resistant strains with CL Brener BZ-susceptible strain showed several single nucleotide polymorphisms, which determined 11 amino acid changes. CL Brener transfected with TcI transporter genes showed 40-47% increased resistance to BZ, whereas no statistical significant increment in drug resistance was observed when CL Brener was transfected with the homologous gene. Only in the parasites transfected with TcI genes there was 2-2.6-fold increased abundance of TcABCG1transporter protein. The analysis in wild type strains also suggests that the level of TcABCG1transporter is related to BZ natural resistance. The characteristics of untranslated regions of TcABCG1genes of BZ-susceptible and resistant strains were investigated by computational tools.

The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis

Coagulase-negative staphylococci, particularly Staphylococcus epidermidis, can be regarded as potential reservoirs of resistance genes for pathogenic strains, e.g., Staphylococcus aureus. The aim of this study was to assess the prevalence of different resistance phenotypes to macrolide, lincosamide, and streptogramins B (MLSB) antibiotics among erythromycin-resistant S. epidermidis, together with the evaluation of genes promoting the following different types of MLSB resistance:ermA, ermB, ermC,msrA, mphC, and linA/A’. Susceptibility to spiramycin was also examined. Among 75 erythromycin-resistantS. epidermidis isolates, the most frequent phenotypes were macrolides and streptogramins B (MSB) and constitutive MLSB (cMLSB). Moreover, all strains with the cMLSB phenotype and the majority of inducible MLSB (iMLSB) isolates were resistant to spiramycin, whereas strains with the MSB phenotype were sensitive to this antibiotic. The D-shape zone of inhibition around the clindamycin disc near the spiramycin disc was found for some spiramycin-resistant strains with the iMLSB phenotype, suggesting an induction of resistance to clindamycin by this 16-membered macrolide. The most frequently isolated gene was ermC, irrespective of the MLSB resistance phenotype, whereas the most often noted gene combination wasermC, mphC, linA/A’. The results obtained showed that the genes responsible for different mechanisms of MLSB resistance in S. epidermidis generally coexist, often without the phenotypic expression of each of them.

Fitness cost of resistance to Bacillus thuringiensis in velvetbean caterpillar Anticarsia gemmatalis Hübner (Lepidoptera, Noctuidae)

Selection pressure to obtain resistant genotypes can result in fitness cost. In this study, we report the effects of the selection pressure of a commercial formulation of Bacillus thuringiensis on biological aspects of a Dipel-resistant strain of velvetbean caterpillar, Anticarsia gemmatalis Hübner. Comparisons of Dipel-resistant and susceptible individuals revealed significant differences in pupal weight and larval development time. Both strains (Dipel-resistant and susceptible) were susceptible to Cry1Ac toxin expressed in foliar cotton tissues. Resistant and susceptible strains showed low survival rates of 22.5% and 51.2%, respectively, when fed with Greene diet containing Bt-cotton. Larvae bioassayed after three laboratory generations presented lower survival and less instar numbers than individuals maintained in the laboratory for more than 144 generations. Pupal weight was 9.4% lower and larval development time was 1.9 days longer in the resistant population than in the susceptible strain. Other parameters, such as duration of pupal stage, adult longevity, number of eggs per female, oviposition period, and egg fertility, remained unaffected.

Spatial variability of physical attributes of an alfisol under different hillslope curvatures

The influence of relief forms has been studied by several authors and explains the variability in the soil attributes of a landscape. Soil physical attributes depend on relief forms, and their assessment is important in mechanized agricultural systems, such as of sugarcane. This study aimed to characterize the spatial variability in the physical soil attributes and their relationship to the hillslope curvatures in an Alfisol developed from sandstone and growing sugarcane. Grids of 100 x 100 m were delimited in a convex and a concave area. The grids had a regular spacing of 10 x 10 m, and the crossing points of this spacing determined a total of 121 georeferenced sampling points. Samples were collected to determine the physical attributes related to soil aggregates, porosity, bulk density, resistance to penetration and moisture within the 0-0.2 and 0.2-0.4 m depth. Statistical analyses, geostatistics and Student's t-tests were performed with the means of the areas. All attributes, except aggregates > 2 mm in the 0-0.2 m depth and macroporosity at both depths, showed significant differences between the hillslope curvatures. The convex area showed the highest values of the mean weighted diameter, mean geometric diameter, aggregates > 2 mm, 1-2 mm aggregates, total porosity and moisture and lower values of bulk density and resistance to penetration in both depth compared to the concave area. The number of soil attributes with greater spatial variability was higher in the concave area.

Physical properties and organic carbon content of a Rhodic Kandiudox fertilized with pig slurry and poultry litter

The impact of pig slurry and poultry litter fertilization on soils depends on the conditions of use and the amounts applied. This study evaluated the effect of organic fertilizers after different application periods in different areas on the physical properties and organic carbon contents of a Rhodic Kandiudox, in Concordia, Santa Catarina, in Southern Brazil. The treatments consisted of different land uses and periods of pig and poultry litter fertilization: silage maize (M7 years), silage maize (M20 years), annual ryegrass pasture (P3 years), annual ryegrass pasture (P15 years), perennial pasture (PP20 years), yerba mate tea (Mt20 years), native forest (NF), and native pasture without manure application (P0). The 0-5, 5-10 and 10-20 cm soil layers were sampled and analyzed for total organic carbon, total nitrogen and soil physical properties such as density, porosity, aggregation, degree of flocculation, and penetration resistance. The organic carbon levels in the cultivated areas treated with organic fertilizer were even lower than in native forest soil. The organic fertilizers and studied management systems reduced the flocculation degree of the clay particles, and low macroporosity was observed in some areas. Despite these changes, a good soil physical structure was maintained, e.g., soil density and resistance to penetration were below the critical limits, whereas aggregate stability was high, which is important to reduce water erosion in these areas with rugged terrain in western Santa Catarina, used for pig and poultry farming.

Effects of tillage systems on physical properties of a cohesive yellow argisol in the northern state of Espírito Santo, Brazil

Tillage systems are a key element of the technology of crop production, both with a view to crop yield and from the perspective of soil conservation and sustainability of the production system. The aim of this paper was to evaluate the effects of five tillage systems on the physical properties of a cohesive Yellow Argisol. The experiment was installed in the field on January 21, 2011 and lasted 260 days, in an area previously used as pasture with Brachiaria grass without liming or fertilization, but irrigated by a low pressure spray system. The treatments, in five replications and in a randomized block design, consisted of: 1) disk plow (twice) + disk harrow + ridge-furrow tillage (raising a ridge along the planting row), 135 days after transplanting (DP + RID); 2) disk plow (twice) + disk harrow (DP no RID); 3) subsoiler (SB); 4) disk plow (twice) + disk harrow + scarification with three shanks along the plant row (DP + SPR); and 5) disk plow (twice) + disk harrow + scarification with three shanks in the total area (DP + STA). In all tillage systems, furrows were mechanically opened for the papaya plants. After the treatments, the mechanical resistance to penetration was determined, followed by soil moisture, mean weight diameter (MWD), geometric mean diameter (GMD), bulk density (BD), macroporosity (Ma), microporosity (Mi), and number of fruits per plant. There were differences in penetration resistance (PR) between treatments. The subsoiler was more effective to decrease RP to a distance of 0.35 m from the plants, perpendicular to the plant row. The scarifier resulted in a lower PR than DP or SB, even at the depth of 0.40 m, and it was more effective at greater distances perpendicular to the plant. All tillage systems induced a PR between 2.0 and 3.0 MPa at the depth with the highest concentration of papaya tree roots (0-0.25 m), improving the physical conditions to this depth. There was no statistical difference among the treatments for BD, Ma, Mi, MWD, and GMD at a depth of 0.20 m. The disk plow changed the physical properties of the soil most intensely to a depth of 0.20 m. The use of scarification, reduced tillage with a forest subsoiler, or ridge-furrow tillage did not improve the physical properties in the rhizosphere. Reduced tillage with a forest subsoiler resulted in a lower number of fruits per plant than all other treatments, which did not differ from each other.

Evaluation of peanut genotypes for resistance to Tomato spotted wilt virus by mechanical and thrips inoculation

The objective of this work was to evaluate the reactions of three peanut breeding lines (IC-10, IC-34, and ICGV 86388) to Tomato spotted wilt virus (TSWV) by mechanical and thrips inoculation, under greenhouse conditions, and compare them to the reactions of cultivars SunOleic, Georgia Green, and the breeding line C11-2-39. TSWV infection by mechanical inoculation was visually assessed using an index ranging from 0 (no symptoms) to 4 (apical death). Enzyme-linked immunosorbent assay was used to confirm TSWV infection from both mechanical and thrips inoculations. IC-10, IC-34, ICGV 86388, and C11-2-39 were more resistant than the cultivars SunOleic and Georgia Green based on mechanical inoculation. Upon thrips inoculation only IC-34 and ICGV-86388 were infected by TSWV, as demonstrated by reverse transcription polymerase chain reaction (RT-PCR), although no symptoms of infection were observed. The peanut breeding lines IC-10, IC-34, and ICGV 86388 show higher level of resistance to TSWV than cultivar Georgia Green considered a standard for TSWV resistance.

Validation of microsatellite markers for assisted selection of soybean resistance to cyst nematode races 3 and 14

The objective of this work was to validate microsatellite markers associated with resistance to soybean cyst nematode (Heterodera glycines Ichinohe) races 3 and 14, in soybean (Glycine max L.) genotypes, for use in marker-assisted selection (MAS) programs. Microsatellites of soybean linkage groups A2, D2 and G were tested in two populations, and their selection efficiencies were determined. The populations were 65 F2:3 families from Msoy8001 (resistant) x Conquista (susceptible) cross, and 66 F2:3 families of S5995 (resistant) x Renascença (susceptible) cross, evaluated for resistance to races 3 and 14, respectively. Families with female index up to 30% were considered moderately resistant. Markers of A2 and G linkage groups were associated with resistance to race 3. Markers Satt309 and GMENOD2B explained the greatest proportion of phenotypic variance in the different groups. The combinations Satt309+GMENOD2B and Satt309+Satt187 presented 100% selection efficiency. Resistance to race 14 was associated with markers of G linkage group, and selection efficiency in the Satt309+Satt356 combination was 100%. The selection differential obtained by phenotypic and marker assisted selection showed that both can result in similar gains.

Inheritance pattern and selection criteria for resistance to soybean cyst nematode races 3 and 9

The objective of this work was to determine soybean resistance inheritance to Heterodera glycines Ichinohe (soybean cyst nematode - SCN) races 3 and 9, as well as to evaluate the efficiency of direct and indirect selection in a soybean population of 112 recombinant inbred lines (RIL) derived from the resistant cultivar Hartwig. The experiment was conducted in a completely randomized design, in Londrina, PR, Brazil. The estimated narrow-sense heritabilities for resistance to races 3 and 9 were 80.67 and 77.97%. The genetic correlation coefficient (r g = 0.17; p<0.01) shows that some genetic components of resistance to these two races are inherited together. The greatest genetic gain by indirect selection was obtained to race 9, selecting to race 3 due to simpler inheritance of resistance to race 9 and not because these two races share common resistance genes. The resistance of cultivar Hartwig to races 3 and 9 is determined by 4 and 2 genes, respectively. One of these genes confers resistance to both races, explaining a fraction of the significant genetic correlation found between resistance to these SCN races. The inheritance pattern described indicates that selection for resistance to SCN must be performed for each race individually.

Inheritance of resistance to soybean cyst nematode races 3 and 14 in soybean RIL and F2 populations

The objective of this work was to evaluate the soybean inheritance of resistance to cyst nematode races 3 and 14. The following populations where evaluated: one population of recombinant inbred lines (RILs) [Hartwig (resistant) x Y23 (susceptible line)] for races 3, 14 and 9; one population of families F2:3 [M-SOY 8001 (resistant) x MB/BR 46 - Conquista (susceptible)] for race 3; and one population of families F2:3 [(S5995 (resistant) x BRSMG Renascença (susceptible)] for race 14. In RIL populations, four epistatic genes were identified which conditioned resistance to race 14, and three epistatic ones for resistance to races 3 and 9. The lack of one gene provided moderate resistance under all situations. The highest number of genes for resistance to race 14 points out that genes responsible for lower effects might be involved. In population F2:3 from M-SOY 8001 x MB/BR 46 - Conquista, one recessive gene for moderate resistance and two recessive genes complete resistance to race 3 were identified. Two recessive genes conditioning moderate resistance to race 14 were identified in population F2:3 from the crossing S5995 x BRSMG Renascença. These results will be useful in designing crossings, involving these parentals, with higher possibility to accumulating genes that provide resistance to several SCN races.