Schistosoma mansoni: a method for inducing resistance to praziquantel using infected Biomphalaria glabrata snails

To elucidate the mechanisms of antischistosoma resistance, drug-resistant Schistosoma mansoni laboratory isolates are essential. We developed a new method for inducing resistance to praziquantel (PZQ) using successive drug treatments of Biomphalaria glabrata snails infected with S. mansoni. Infected B. glabrata were treated three times with 100 mg/kg PZQ for five consecutive days with a one-week interval between them. After the treatment, the cercariae (LE-PZQ) produced from these snails and the LE strains (susceptible) were used to infect mice. Forty-five days after infection, mice were treated with 200, 400 or 800 mg/kg PZQ. Thirty days post-treatment, we observed that the mean number of worms recovered by perfusion was significantly higher in the group of mice infected with the LE-PZQ isolate treated with 200 and 400 mg/kg in comparison to the LE strain with the same treatment. Moreover, there was a significant difference between the ED50 (effective dose required to kill 50% of the worms) of the LE-PZQ isolate (362 mg/kg) and the LE strain (68 mg/kg). In the in vitro assays, the worms of the LE-PZQ isolate were also less susceptible to PZQ. Thus, the use of infected snails as an experimental model for development of resistance to S. mansoni is effective, fast, simple and cheap.

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.

Structural quality of soils cultivated with coffee and pasture in an environmental protection area

Studies of soils in Environmental Protection Areas (EPAs) are of great importance, because they are an essential component of ecosystems, directly interfering in environmental sustainability. The objective of this study was to evaluate the structural quality of soil cultivated with coffee and used as pasture in the Capituva's River microbasin, which is located in the Environmental Protection Area in Coqueiral, south of the state of Minas Gerais. Uniaxial compression test (preconsolidation test) and soil resistance to penetration were used. Undisturbed samples were taken from the surface layer (0-5 cm) of the soils in the area: a typic dystrophic Red Latosol (LVd - Oxisol), a typic eutrophic Red Argisol (PVe - Ultisol), and a typic dystrophic Haplic Cambisol (CXbd - Inceptisol). A significant linear positive correlation was observed between the results of the preconsolidation test and soil resistance to penetration. Load bearing capacity of soil could be estimated accordingly by means of penetration resistance for LVd, PVe, and CXbd. Cambisol - CXbd showed lower loading support capacity and resistance to penetration than LVd and PVe, due to the better crop management in this soil that resulted in higher physical quality which accounts for higher production and environmental sustainability.

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.

Effect of cropping systems in no-till farming on the quality of a Brazilian Oxisol

The no-till system with complex cropping sequences may improve the structural quality and carbon (C) sequestration in soils of the tropics. Thus, the objective of this study was to evaluate the effects of cropping sequences after eight years under the no-till system on the physical properties and C sequestration in an Oxisol in the municipality of Jaboticabal, Sao Paulo, Brazil. A randomized split-block design with three replications was used. The treatments were combinations of three summer cropping sequences - corn/corn (Zea mays L.) (CC), soybean/soybean (Glycine max L. Merryll) (SS), and soybean-corn (SC); and seven winter crops - corn, sunflower (Helianthus annuus L.), oilseed radish (Raphanus sativus L.), pearl millet (Pennisetum americanum (L.) Leeke), pigeon pea (Cajanus cajan (L.) Millsp), grain sorghum (Sorghum bicolor (L.) Moench), and sunn hemp (Crotalaria juncea L.). Soil samples were taken at the 0-10 cm depth after eight years of experimentation. Soil under SC and CC had higher mean weight diameter (3.63 and 3.55 mm, respectively) and geometric mean diameter (3.55 and 2.92 mm) of the aggregates compared to soil under SS (3.18 and 2.46 mm). The CC resulted in the highest soil organic C content (17.07 g kg-1), soil C stock (15.70 Mg ha-1), and rate of C sequestration (0.70 Mg ha-1 yr-1) among the summer crops. Among the winter crops, soil under pigeon pea had the highest total porosity (0.50 m³ m-3), and that under sunn hemp had the highest water stable aggregates (93.74 %). In addition, sunn hemp did not differ from grain sorghum and contained the highest soil organic C content (16.82 g kg-1) and also had the highest rate of C sequestration (0.67 Mg ha-1 yr-1). The soil resistance to penetration was the lower limit of the least limiting water range, while the upper limit was air-filled porosity for soil bulk densities higher than 1.39 kg dm-3 for all cropping sequences. Within the SC sequence, soil under corn and pigeon pea increased least limiting water range by formation of biopores because soil resistance to penetration decreased with the increase in soil bulk density.

Quantification of the least limiting water range in an oxisol using two methodological strategies

The least limiting water range (LLWR) has been used as an indicator of soil physical quality as it represents, in a single parameter, the soil physical properties directly linked to plant growth, with the exception of temperature. The usual procedure for obtaining the LLWR involves determination of the water retention curve (WRC) and the soil resistance to penetration curve (SRC) in soil samples with undisturbed structure in the laboratory. Determination of the WRC and SRC using field measurements (in situ ) is preferable, but requires appropriate instrumentation. The objective of this study was to determine the LLWR from the data collected for determination of WRC and SRC in situ using portable electronic instruments, and to compare those determinations with the ones made in the laboratory. Samples were taken from the 0.0-0.1 m layer of a Latossolo Vermelho distrófico (Oxisol). Two methods were used for quantification of the LLWR: the traditional, with measurements made in soil samples with undisturbed structure; and in situ , with measurements of water content (θ), soil water potential (Ψ), and soil resistance to penetration (SR) through the use of sensors. The in situ measurements of θ, Ψ and SR were taken over a period of four days of soil drying. At the same time, samples with undisturbed structure were collected for determination of bulk density (BD). Due to the limitations of measurement of Ψ by tensiometer, additional determinations of θ were made with a psychrometer (in the laboratory) at the Ψ of -1500 kPa. The results show that it is possible to determine the LLWR by the θ, Ψ and SR measurements using the suggested approach and instrumentation. The quality of fit of the SRC was similar in both strategies. In contrast, the θ and Ψ in situ measurements, associated with those measured with a psychrometer, produced a better WRC description. The estimates of the LLWR were similar in both methodological strategies. The quantification of LLWR in situ can be achieved in 10 % of the time required for the traditional method.

ASPECTS OF THE SILVOPASTORAL SYSTEM CORRELATED WITH PROPERTIES OF A TYPIC QUARTZIPSAMMENT (ENTISOL) IN MATO GROSSO DO SUL, BRAZIL

In Brazil, grazing mismanagement may lead to soil and pasture degradation. To impede this process, integrated cropping systems such as silvopasture have been an effective alternative, allied with precision agriculture based on soil mapping for site-specific management. In this study, we aimed to define the soil property that best sheds light on the variability of eucalyptus and forage yield. The experiment was conducted in the 2011/12 crop year in Ribas do Rio Pardo, Mato Grosso do Sul State, Brazil. We analyzed linear and spatial correlations between eucalyptus traits and physical properties of a Typic Quartzipsamment at two depths (0.00-0.10 and 0.10-0.20 m). For that purpose, we set up a geostatistical grid for collection at 72 points. Gravimetric moisture in the 0.00-0.10 m layer is an important index of soil physical quality, showing correlation to eucalyptus circumference at breast height (CBH) in a Typic Quartzipsamment. With an increase in resistance to penetration in the soil surface layer, there is an increase in eucalyptus height and in neutral detergent fiber content in the forage crop. From a spatial point of view, the height of eucalyptus and the neutral detergent fiber of forage can be estimated by co-kriging analysis with soil resistance to penetration. Resistance to penetration values above 2.3 MPa indicated higher yielding sites.