Mosquito traps designed to capture Aedes aegypti (Diptera: Culicidae) females: preliminary comparison of Adultrap, MosquiTRAP and backpack aspirator efficiency in a dengue-endemic area of Brazil

In this report, the efficiency of Adultrap under field conditions is compared to a CDC backpack aspirator and to MosquiTRAP. An urban dengue-endemic area of Rio de Janeiro was selected to evaluate the efficiency of mosquito traps in capturing Aedes aegypti females. Adultrap and aspirator captured similar numbers of Ae. aegypti females, with the former showing high specificity to gravid individuals (93.6%). A subsequent mark-release-recapture experiment was conducted to evaluate Adultrap and MosquiTRAP efficiency concomitantly. With a 6.34% recapture rate, MosquiTRAP captured a higher mean number of female Ae. aegypti per trap than Adultrap (Ç2 = 14.26; df = 1; p < 0,05). However, some MosquiTRAPs (28.12%) contained immature Ae. aegypti after 18 days of exposure in the field and could be pointed as an oviposition site for female mosquitoes. Both trapping methods, designed to collect gravid Ae. aegypti females, seem to be efficient, reliable and may aid routine Ae. aegypti surveillance.

Uptake of macromolecules by cercariae during skin penetration and transformation to schistosomula (Schistosoma mansoni)

Here, we observed the uptake of membrane-impermeant molecules by cercariae as they penetrate the skin and are transformed into schistosomula. We propose that membrane-impermeant molecules, Lucifer Yellow, Propidium iodide and Hoechst 33258 enter the parasite through both thenephridiopore and the surface membrane and then diffuse throughout the body of the parasite. We present a hypothesis that the internal cells of the body of the schistosomulum represent a new host-parasite interface, at which skin-derived growth factors may stimulate receptors on internal membranes during transformation of the cercariae into the schistosomulum.

Efficiency of diagnostic biomarkers among colonic schistosomiasis Egyptian patients

The schistosomal parasite plays a critical role in the development of malignant lesions in different organs. The pathogenesis of cancer is currently under intense investigation to identify reliable prognostic indices for disease detection. The objective of this paper is to evaluate certain biochemical parameters as diagnostic tools to efficiently differentiate between colonic carcinoma and colonic carcinoma associated with schistosomal infection among Egyptian patients. The parameters under investigation are interleukin 2 (IL-2), tumour necrosis factor alpha (TNF-α), carcinoembryonic antigen (CEA) levels, tissue telomerase, pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD) and lactate dehydrogenase (LDH) enzyme activities. The results revealed a significant elevation in the level of the tumour markers IL-2, TNF-α and CEA as well as the activities of LDH, telomerase and G-6-PD among non-bilharzial and bilharzial colonic cancer groups, with a more potent effect in bilharzial infection-associated colonic cancer. A significant inhibition in PK activity was recorded in the same manner as compared to normal tissues. The efficacy of this biomarker was also evaluated through detecting sensitivity, specificity, negative and positive predictive values. In conclusion, schistosomal colonic carcinoma patients displayed more drastic changes in all parameters under investigation. The combination of the selected parameters succeeded in serving as biomarkers to differentiate between the two malignant types.

Histopathological examination of nerve samples from pure neural leprosy patients: obtaining maximum information to improve diagnostic efficiency

Nerve biopsy examination is an important auxiliary procedure for diagnosing pure neural leprosy (PNL). When acid-fast bacilli (AFB) are not detected in the nerve sample, the value of other nonspecific histological alterations should be considered along with pertinent clinical, electroneuromyographical and laboratory data (the detection of Mycobacterium leprae DNA with polymerase chain reaction and the detection of serum anti-phenolic glycolipid 1 antibodies) to support a possible or probable PNL diagnosis. Three hundred forty nerve samples [144 from PNL patients and 196 from patients with non-leprosy peripheral neuropathies (NLN)] were examined. Both AFB-negative and AFB-positive PNL samples had more frequent histopathological alterations (epithelioid granulomas, mononuclear infiltrates, fibrosis, perineurial and subperineurial oedema and decreased numbers of myelinated fibres) than the NLN group. Multivariate analysis revealed that independently, mononuclear infiltrate and perineurial fibrosis were more common in the PNL group and were able to correctly classify AFB-negative PNL samples. These results indicate that even in the absence of AFB, these histopathological nerve alterations may justify a PNL diagnosis when observed in conjunction with pertinent clinical, epidemiological and laboratory data.

The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

Efficiency of the induced mating technique for Toxorhynchites theobaldi (Diptera, Culicidae)

Toxorhynchites mosquitoes play important ecological roles in aquatic microenvironments, and are frequently investigated as potential biological control agents of mosquito disease vectors. Establishment of Toxorhynchites laboratory colonies can be challenging because for some species, mating and insemination either do not occur or require a prohibitive amount of laboratory space for success. Consequently, artificial insemination techniques have been developed to assist with mass rearing of these species. Herein we describe an adapted protocol for colony establishment of T. theobaldi, a species with broad distribution in the Neotropics. The success of the technique and its implications are discussed.

Influence of soil pH on inorganic phosphorus sorption and desorption in a humid brazilian Ultisol

Liming is a common practice to raise soil pH and increase phosphorus (P) bioavailability in tropical regions. However, reports on the effect of liming on P sorption and bioavailability are controversial. The process of phosphorus desorption is more important than P sorption for defining P bioavailability. However few studies on the relationship between soil pH and P desorption are available, and even fewer in the tropical soils. The effects of soil pH on P sorption and desorption in an Ultisol from Bahia, Brazil, were investigated in this study. Phosphorus sorption decreased by up to 21 and 34 % with pH increases from 4.7 to 5.9 and 7.0, respectively. Decreasing Langmuir K parameter and decreasing partition coefficients (Kd) with increasing pH supported this trend. Phosphorus desorption was positively affected by increased soil pH by both the total amount of P desorbed and the ratio of desorbed P to initially sorbed P. A decreased K parameter and increased Kd value, particularly at the highest pH value and highest P-addition level, endorsed this phenomenon. Liming the soil had the double effect of reducing P sorption (up to 4.5 kg ha-1 of remaining P in solution) and enhancing P desorption (up to 2.7 kg ha-1 of additionally released P into solution).

Leaf senescence of common bean plants as affected by soil phosphorus supply

Responses of leaf senescence to P supply could constitute adaptive mechanisms for plant growth under P-limiting conditions. The aim of this study was to evaluate the effects of soil P supply on leaf senescence of common bean (Phaseolus vulgaris L.). Eight P levels, ranging from 5 to 640 mg kg-1 P, were applied to pots containing four bean plants of cultivar Carioca in 10 kg of an Oxic Haplustult soil. Attached leaves were counted weekly, abscised leaves were collected every other day, and seeds were harvested at maturity. The number of live leaves increased until 48 days after emergence (DAE) and decreased afterwards, irrespective of applied P levels. At lower applied P levels, the initial increase and the final decrease of leaf number was weak, whereas at higher applied P levels the leaf number increased intensively at the beginning of the growth cycle and decreased strongly after 48 DAE. Dry matter and P accumulated in senesced leaves increased as soil P levels increased until 61 DAE, but differences between P treatments narrowed thereafter. The greatest amounts of dry mass and P deposited by senesced leaves were observed at 48-54 DAE for high P levels, at 62-68 DAE for intermediate P levels and at 69-76 DAE for low P levels. These results indicate that soil P supply did not affect the stage of maximal leaf number and the beginning of leaf senescence of common bean plants, but the stage of greatest deposition of senesced leaves occurred earlier in the growth cycle as the soil P supply was raised.

Relationships between grain yield and accumulation of biomass, nitrogen and phosphorus in common bean cultivars

Shoot biomass is considered a relevant component for crop yield, but relationships between biological productivity and grain yield in legume crops are usually difficult to establish. Two field experiments were carried out to investigate the relationships between grain yield, biomass production and N and P accumulation at reproductive stages of common bean (Phaseolus vulgaris) cultivars. Nine and 18 cultivars were grown on 16 m² plots in 1998 and 1999, respectively, with four replications. Crop biomass was sampled at four growth stages (flowering R6, pod setting R7, beginning of pod filling R8, and mid-pod filling R8.5), grain yield was measured at maturity, and N and P concentrations were determined in plant tissues. In both years, bean cultivars differed in grain yield, in root mass at R6 and R7 stages, and in shoot mass at R6 and R8.5, whereas at R7 and R8 differences in shoot mass were significant in 1998 only. In both years, grain yield did not correlate with shoot mass at R6 and R7 and with root mass at R6. Grain yield correlated with shoot mass at R8 in 1999 but not in 1998, with shoot mass at R8.5 and with root mass at R7 in both years. Path coefficient analysis indicated that shoot mass at R8.5 had a direct effect on grain yield in both years, that root mass at R7 had a direct effect on grain yield in 1998, and that in 1999 the amounts of N and P in shoots at R8.5 had indirect effects on grain yield via shoot mass at R8.5. A combined analysis of both experiments revealed that biomass accumulation, N and P in shoots at R6 and R7 as well as root mass at R6 were similar in both years. In 1998 however bean accumulated more root mass at R7 and more biomass and N and P in shoots at R8 and R8.5, resulting in a 57 % higher grain yield in 1998. This indicates that grain yield of different common bean cultivars is not intrinsically associated with vegetative vigor at flowering and that mechanisms during pod filling can strongly influence the final crop yield. The establishment of a profuse root system during pod setting, associated with the continuous N and P acquisition during early pod filling, seems to be relevant for higher grain yields of common bean.

Soil phosphorus fractions in sandy soils amended with cattle manure for long periods

Phosphorus fractions were determined in soil samples from areas fertilized or not with farmyard cattle manure (FYM) and in samples of FYM used in the semi-arid region of Paraiba state, Brazil. Soil samples were taken from the 0-20; 20-40 and 40-60 cm layers of 18 cultivated areas, which, according to interviews with farmers, had been treated with 12 to 20 t ha-1 FYM annually, for the past 2 to 40 years. Soil samples were also collected from four unfertilized pasture areas as controls. Phosphorus in the soil samples was sequentially extracted with water (Pw), resin (Pres), NaHCO3 (Pi bic and Po bic), NaOH (Pi hid and Po hid), H2SO4 (Pacid) and, finally, by digestion with H2SO4/H2O2 (Presd). Nine FYM samples were extracted with water, resin, Mehlich-1, H2SO4, NaOH or digestion with H2SO4/H2O2, not sequentially, and the extracts analyzed for P. The sampled areas had homogeneous, sandy and P-deficient soils; increases in total soil P (Pt) above the mean value of the control areas (up to 274 mg kg-1 in the 0-20 cm layer of the most P-enriched samples) were therefore attributed to FYM applications, which was the only external P input in the region. Regression analysis was used to study the relationship between soil P fractions and Pt. The Pacid fraction, related to Ca-P forms, showed the greatest increases (p < 0.01) as a result of FYM applications, rising from 8.4 mg kg-1 in a non-fertilized sample to 43.8 mg kg-1 in the sample with the highest Pt content. The sum of Pw, Pres and Pi bic, considered as labile P, showed comparable increases with Pacid, while Pi hid showed the smallest increase due to FYM applications. Organic P forms also increased, more so the fraction Po hid, considered less labile, than the more labile one, Po bic. The residual P fraction was practically half of Pt, independently of the Pt value. Increases in labile P, Pacid and organic P were justified by the high average concentration of Pw (36 %), Pacid (34 %), and Po hid (30 %) in the FYM. Significant changes in the proportion of P forms among soil layers indicated the downward movement of P in organic forms.

Sewage sludge as nitrogen and phosphorus source for cane-plant and first ratoon crops

The use of sewage sludge in Brazilian agriculture was regulated by the resolution no. 375 Conama, in 2006. However, there is a lack of research to adequate the mineral N and P fertilizer doses to be applied in agricultural fields treated with this residue. In a field experiment, the effects of application rates of sewage sludge and mineral N and P fertilizers on the productivity and technical characteristics of the cane-plant and first ratoon (residual effect) crops were evaluated. Four doses of sewage sludge (0, 3.6, 7.2 and 10.8 t ha-1, dry base), of N (0, 30, 60 and 90 kg ha-1) and of P2O5 (0, 60, 120 and 180 kg ha-1) were combined in a factorial and laid out on randomized block design, a with two replications. To evaluate the residual effect of the sludge, 120 kg ha-1 N and 140 kg ha-1 of K2O were applied in all plots. Sludge application at cane planting, with or without N and/or P fertilizer increased the stalk yield from 84 up to 118 t ha-1, with no alteration in the sugarcane quality, compared with the application of NPK fertilizer alone, resulting in a stalk yield of 91 t ha-1. The study of the response surface for stalk yield on lowfertility soil was the basis for a recommendation of mineral N and P fertilizer doses for sugarcane implantation as related to sewage sludge application rates. It was also concluded that a sludge application of 10.8 t ha-1, which is the sludge dose established based on the N criterion according to the resolution Conama nº 375, could a) reduce the use of mineral N by 100 % and of P2O5 by 30 %, with increments of 22 % in stalk yield, as a direct effect of sludge application to cane plant crop, and b) increase the stalk yield in the second harvest (first ratoon) by up to 12 % and sugar yield by up to 11 %, by the residual effect of sludge application to sugar cane.

Effects of arbuscular mycorrhizal fungi and phosphorus fertilization on post vitro growth of micropropagated Zingiber officinale roscoe

The rhizomes of Zingiber officinale Roscoe (ginger) are widely used for their medicinal and flavoring properties, whereas the influence of root symbionts on their growth is poorly understood. In this study, the effects of phosphate fertilization and inoculation with a mixture of arbuscular mycorrhizal fungi (AMF) (isolates Glomus clarum RGS101A, Entrophospora colombiana SCT115A and Acaulospora koskei SPL102A) on survival, growth and development of micropropagated ginger were investigated. After transplanting to post vitro conditions, the ginger microplants were subjected to the following treatments: a) AMF mixture, b) P addition (25 mg kg-1), c) AMF + P, and d) non-mycorrhizal control without P addition. After eight months of growth, survival ranged from 86 to 100 % in the AMF and AMF+P treatments versus 71 % survival in control and P treatments. In the AMF, P and AMF+P treatments, the shoot, root and rhizome biomass production were significantly larger than in the control plants. In the non-mycorrhizal control plants the leaf number, leaf area, number of shoots/plants, and shoot length were significantly lower than in the AMF, P and AMF+P treatments. Root colonization ranged from 81 to 93 % and was not affected by P application. The data confirmed the response of several growth variables of micropropagated ginger to mycorrhizal colonization and P addition.

Seeds enriched with phosphorus and molybdenum improve the contribution of biological nitrogen fixation to common bean as estimated by 15n isotope dilution

Seeds with a high concentration of P or Mo can improve the growth and N accumulation of the common bean (Phaseolus vulgaris L.), but the effect of enriched seeds on biological N2 fixation has not been established yet. This study aimed to evaluate the effect of seeds enriched with P and Mo on growth and biological N2 fixation of the common bean by the 15N isotope dilution technique. An experiment was carried out in pots in a 2 x 3 x 2 x 2 factorial design in randomized blocks with four replications, comprising two levels of soil applied P (0 and 80 mg kg-1), three N sources (without N, inoculated with rhizobia, and mineral N), two seed P concentrations (low and high), and two seed Mo concentrations (low and high). Non-nodulating bean and sorghum were used as non-fixing crops. The substrate was 5.0 kg of a Red Latosol (Oxisol) previously enriched with 15N and mixed with 5.0 kg of sand. Plants were harvested 41 days after emergence. Seeds with high P concentration increased the growth and N in shoots, particularly in inoculated plants at lower applied P levels. Inoculated plants raised from high P seeds showed improved nodulation at both soil P levels. Higher soil P levels increased the percentage of N derived from the atmosphere (%Ndfa) in bean leaves. Inoculation with the selected strains increased the %Ndfa. High seed P increased the %Ndfa in inoculated plants at lower soil P levels. High seed Mo increased the %Ndfa at lower soil P levels in plants that did not receive inoculation or mineral N. It is concluded that high seed P concentration increases the growth, N accumulation and the contribution of the biological N2 fixation in the common bean, particularly in inoculated plants grown at lower soil P availability.

Diversity and efficiency of bradyrhizobium strains isolated from soil samples collected from around sesbania virgata roots using cowpea as trap species

The genetic diversity of ten Bradyrhizobium strains was evaluated for tolerance to high temperatures, to different salinity levels and for the efficiency of symbiosis with cowpea plants (Vigna unguiculata (L.) Walp.). Eight of these strains were isolated from nodules that appeared on cowpea after inoculation with suspensions of soil sampled from around the root system of Sesbania virgata (wand riverhemp) in ecosystems of South Minas Gerais. The other two strains used in our analyses as references, were from the Amazon and are currently recommended as cowpea inoculants. Genetic diversity was analyzed by amplifying repetitive DNA elements with the BOX primer, revealing high genetic diversity with each strain presenting a unique band profile. Leonard jar assays showed that the strains UFLA 03-30 and UFLA 03-38 had the highest N2-fixing potentials in symbiosis with cowpea. These strains had more shoot and nodule dry matter, more shoot N accumulation, and a higher relative efficiency than the strains recommended as inoculants. All strains grew in media of pH levels ranging from 4.0 to 9.0. The strains with the highest N2-fixing efficiencies in symbiosis with cowpea were also tolerant to the greatest number of antibiotics. However, these strains also had the lowest tolerance to high salt concentrations. All strains, with the exceptions of UFLA 03-84 and UFLA 03-37, tolerated temperatures of up to 40 ºC. The genetic and phenotypic characteristics of the eight strains isolated from soils of the same region were highly variable, as well as their symbiotic efficiencies, despite their common origin. This variability highlights the importance of including these tests in the selection of cowpea inoculant strains.

Mobility of inorganic and organic phosphorus forms under different levels of phosphate and poultry litter fertilization in soils

The eutrophication of aquifers is strongly linked to the mobility of P in soils. Although P mobility was considered irrelevant in a more distant past, more recent studies have shown that P, both in organic (Po) and inorganic forms (Pi), can be lost by leaching and eluviation through the soil profile, particularly in less weathered and/or sandier soils with low P adsorption capacity. The purpose of this study was to determine losses of P forms by leaching and eluviation from soil columns. Each column consisted of five PVC rings (diameter 5 cm, height 10 cm), filled with two soil types: a clayey Red-Yellow Latosol and a sandy loam Red-Yellow Latosol, which were exposed to water percolation. The soils were previously treated with four P rates (as KH2PO4 ) to reach 0, 12.5, 25.0 and 50 % of the maximum P adsorption capacity (MPAC). The P source was homogenized with the whole soil volume and incubated for 60 days. After this period the soils were placed in the columns; the soil of the top ring was mixed with five poultry litter rates of 0, 20, 40, 80, and 160 t ha-1 (dry weight basis). Treatments consisted of a 4 x 5 x 2 factorial scheme corresponding to four MPAC levels, five poultry litter rates, two soils, with three replications, arranged in a completely randomized block design. Deionized water was percolated through the columns 10 times in 35 days to simulate about 1,200 mm rainfall. In the leachate of each column the inorganic P (reactive P, Pi) and organic P forms (unreactive P, Po) were determined. At the end of the experiment, the columns were disassembled and P was extracted with the extractants Mehlich-1 (HCl 0.05 mol L-1 and H2SO4 0.0125 mol L-1) and Olsen (NaHCO3 0.5 mol L-1; pH 8.5) from the soil of each ring. The Pi and Po fractions were measured by the Olsen extractant. It was found that under higher poultry litter rates the losses of unreactive P (Po) were 6.4 times higher than of reactive P (Pi). Both the previous P fertilization and increasing poultry litter rates caused a vertical movement of P down the soil columns, as verified by P concentrations extracted by Mehlich-1 and NaHCO3 (Olsen). The environmental critical level (ECL), i.e., the P soil concentration above which P leaching increases exponentially, was 100 and 150 mg dm-3 by Mehlich-1 and 40 and 60 mg dm-3 by Olsen, for the sandy loam and clay soils, respectively. In highly weathered soils, where residual P is accumulated by successive crops, P leaching through the profile can be significant, particularly when poultry litter is applied as fertilizer.