Biology of Amblyomma aureolatum (Pallas, 1772) (Acari: Ixodidae) on some laboratory hosts in Brazil

The ixodid Amblyomma aureolatum is suspected to play a role in the epidemiology of wild life-cycle hemoparasites, which frequently infect dogs in rural and hunting areas in Brazil. Little is known about its bionomics. The objective of the present study was to evaluate some bionomic aspects of A. aureolatum ticks in Brazil. One engorged female, collected from a dog (Canis familiaris) in São Sebastião das Águas Claras, State of Minas Gerais, was used to establish a colony in the laboratory. Subsequently its parasitic stage progeny were fed on domestic dogs and laboratory animals. The free-living stages were incubated at 27ºC ± 2°C and minimum 70% relative humidity in a BOD incubator. The egg incubation period ranged from 31 to 34 days; the parasitic period of larvae ranged from 4 to 6 days and ecdysis to nymphs occurred from day 19 up to day 22. The parasitic period of nymphs ranged from 5 to 8 days and the period of ecdysis to adults from 31 to 33 days. The parasitic period of adults ranged from 11 to 15 days, the pre-oviposition period from 6 to 12 days, and the oviposition period from 9 to 38 days. The total duration of the life cycle ranged from 116 to 168 days.

Drug susceptibility testing of Mycobacterium tuberculosis by the broth microdilution method with 7H9 broth

In this study, we have evaluated the broth microdilution method (BMM) for susceptibility testing of Mycobacterium tuberculosis. A total of 43 clinical isolates of M. tuberculosis and H37Rv as a control strain were studied. All isolates were tested by the proportion method and the BMM for isoniazid (INH), rifampicin (RIF), streptomycin (STR), and ethambutol (ETM). The proportion method was carried out according to the National Committee for Clinical Laboratory Standards (NCCLS) on Löwenstein-Jensen (LJ) medium. The BMM was carried out using 7H9 broth with 96 well-plates. All strains were tested at 3.2-0.05 µg/ml, 16-0.25 µg/ml, 32-0.5 µg/ml, and 32-0.5 µg/ml concentrations for INH, RIF, STR, and ETM, respectively. When the BMM was compared with the proportion method, sensitivity was 100, 100, 96.9, and 90.2%, while specificity was 100, 85.7, 90.9, and 100% for INH, RIF, STR, and ETM, respectively. The plates were examined 7, 10, 14, and 21 days after incubation. The majority of the result were obtained at 14th days after incubation, while the proportion method result were ended in 21-28 days. According to our results, it may be suggested that the BMM is suitable for early determining of multidrug-resistance-M. tuberculosis strains in developed or developing countries.

Assessment of schistosomiasis, through school surveys, in the Forest Zone of Pernambuco, Brazil

This work had the objective of assessing the present epidemiological situation regarding schistosomiasis through performing Kato-Katz coproscopic tests on representative samples of schoolchildren from each of the 43 municipality of endemic area of the state of Pernambuco, Brazil. The methodology is recommended by the World Health Organization to conduct sampled surveys among children at elementary school levels, ideal target group for baseline surveys: (i) schools are accessible; (ii) the greatest prevalence of schistosomiasis is found within this group; (iii) the data gathered from this age group can be used for intervention within the community as a whole. The following infection indicators were utilized: positivity (percentage of individuals examined with eggs of Schistosoma mansoni in the feces) and severity (geometric mean number of eggs per gram of feces, epg). These indicators allowed the area in general and the municipalities in particular to be categorized into prevalence and severity classes for S. mansoni. The prevalence classes were: low (<10%), medium (> 10 and < 50%), and high (> 50%); the severity classes were: low (1-99 epg), moderate (100-399 epg), and severe (> 400 epg). For the geohelminthic diseases, the following indicators were used: positivity for each geohelminth (percentage of individuals examined with eggs of geohelminths), and cumulative positivity (percentage of individuals examined with eggs of at least one geohelminth). The municipalities were categorized by means of their cumulative positivity into the following geohelminth prevalence classes (WHO 2002): low (< 50%), medium (> 50 and < 70%), and high (> 70%). The study covered 271 schools in 179 different localities, thus giving a total of 11,234 examinations performed. The overall positivity for S. mansoni was 14.4% and the egg count for this parasite in the feces gave a geometric mean of 67.9 epg which suggests a low general state of infection. These results allow this mesoregion to be categorized as presenting medium prevalence and low severity of schistosomiasis. The overall positivity rates for the geohelminths, Ascaris lumbricoides, Ancylostomidae, and Trichuris trichiura were, respectively, 30.4, 10.1, and 27.8%; the cumulative positivity was 45.4%. These results allow this mesoregion to be categorized as presenting low prevalence of geohelminthic diseases. The data show some municipalities in Pernambuco with prevalence greater than 20%, while others presented parasite loads greater than 100 epg. These indicators attest to the significant morbidity due to schistosomiasis regarding to the severity of infections established in young populations.

Spoligotyping of clinical Mycobacterium tuberculosis isolates from the state of Minas Gerais, Brazil

We performed spoligotyping on 114 strains of the Mycobacterium tuberculosis (Mtb) complex that had been isolated from patients in Minas Gerais Health Units during 2004. A total of 82/114 (72%) clinical isolates were clustered and 32/114 (28%) were unique. Seven shared types containing nine strains were newly created. A total of nine patterns corresponded to unreported orphan strains, as evaluated against all of the strains recorded in the SITVIT2 proprietary database in the Institut Pasteur de la Guadeloupe. The major clades were composed of isolates that belong to the following genotypes: Latin-America and Mediterranean (63/114, 55.3%) (the ill-defined T superfamily) (12/114, 10.5%), Haarlem (8/114, 7%), X clade (6/114, 5.3%), S clade (3/114, 2.6%) and the East-African Indian and Manu types, each with 1/114 (0.9%) isolates. A considerable number of strains (n = 20, 17.5%) showed patterns that did not fall within any of the previously described major clades. We conclude the bulk of tuberculosis (TB) (92/114, 80.7%) in our location is recent evolutionary strains that belong to the principal genetic groups 2/3. Further studies on epidemiology of TB are required to understand Mtb biodiversity and TB transmission in this region.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Leprosy frequency in the world, 1999-2010

In 1991, the World Health Organization (WHO) committed to reducing the prevalence of leprosy to below 1 in 10,000 inhabitants by 2000. Significant improvements in leprosy control have occurred, but leprosy remains a public health problem in many countries due to its high incidence and rate of transmission. This paper reviews data published by the WHO in the years 2000, 2005 and 2010. These data sets included 148 countries or territories that reported to the WHO at least once. Only four countries reported higher prevalence rates in 2010 than in 2000 and eight reported higher case detection rate (CDR) in 2009 than in 1999. Prevalence rate reductions were greater for the first five-year period examined, while CDR reductions were greater in the second five-year period. Thirty-six countries and territories reported at least one prevalence value higher than 1 per 10,000 inhabitants and 32 reported at least one CDR value higher than 9 per 100,000 inhabitants. A total of 39 countries fit at least one of these criteria and all were located in tropical regions.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

Larval development of Physocephala (Diptera, Conopidae) in the bumble bee Bombus morio (Hymenoptera, Apidae)

Larval development of Physocephala (Diptera, Conopidae) in the bumble bee Bombus morio (Hymenoptera, Apidae). In the summer of 2012, a high incidence of conopid larvae was observed in a sample of female B. morio collected in remaining fragments of semidecidual forest and Cerrado, in the municipality of Sorocaba, state of São Paulo, Brazil. The larval development of conopid flies was studied, beginning at the larval instars (LO to L3) and PUP, until the emergence of the imago under laboratory conditions and inside the host. At the first instar, or LO, the microtype larvae measured less than 1 mm in length. During the transition from L1 to L3, the larvae grew in length. At L3, the larvae doubled their length (4 mm) and then started to develop both in length and width, reaching the PUP stage with 10 mm in length and 7 mm in width. The main characteristic that differentiates L3 from the early instars is the larger body size and the beginning of posterior spiracle development. The development from PUP to puparium took less than 24h. The bees died ten days after the fly oviposition, or just before full PUP development. The early development stages (egg-LO to L1) were critical for larva survival. The pupa was visible between the intersegmental sternites and, 32 days after pupation, a female imago of Physocephala sp. emerged from one bee. The puparium and the fly measured approximately 10 mm in length. In a single day of collection, up to 45% of the bumble bees collected were parasitized by conopid flies.

Cation dynamics in soils with different salinity levels growing irrigated rice

Salinity levels in soils of the Outer Coastal Plain of Rio Grande do Sul, Brazil, can be high, due to excess of Na in the irrigation water, evapotranspiration and soil development from marine sediments. The cultivation of irrigated rice could be an alternative, since ion uptake as well as leaching by the establishment of a water layer could mitigate the effects of soil salinity. This study aimed to evaluate the dynamics of basic cations in the solution of Albaqualf soils with different salinity levels growing irrigated rice. The plow layer contained exchangeable Na percentages (ESP) of 5.6, 9.0, 21.2 and 32.7 %. The plant stand, dry matter, Na, K and Ca + Mg uptake at full flowering and grain yield were evaluated. The levels of Na, K, Ca + Mg and electrical conductivity (EC) in the soil solution were also measured weekly during the rice cycle at four soil depths, in the water layer and irrigation water. The Na, K and Ca + Mg uptake by rice at full flowering was used to estimate ion depletion from the layer under root influence. Soil salinity induced a reduction in the rice stand, especially in the soil with ESP of 32.7 %, resulting in lower cation uptake and very low yield at that site. As observed in the water layer and irrigation water, the Na, K, Ca + Mg and EC levels in the soil solution decreased with time at depths of 5, 10 and 20 cm, regardless of the original soil salinity, showing that cation dynamics in the plow layer was determined by leaching and root uptake, rather than by the effect of evapoconcentration of basic cations in the soil surface layer.

Contribution of macroporosity to water flux of a soil under different tillage systems

In view of the importance of the macroporosity for the water transport properties of soils, its quantitative assessment is a challenging task. Measurements of hydraulic conductivity (K) at different soil water tensions and the quantification of water-conducting macropores (θM) of a soil under different tillage systems could help understand the effects on the soil porous system and related hydraulic properties. The purpose of this study was to assess the effects of Conventional Tillage (CT), Chisel Plow (CP) and No Tillage (NT) on θM and on K; and to quantify the contribution of macroporosity to total water flux in a loam soil. A tension disc infiltrometer was used at two soil water pressure heads (-5 cm, and 0) to infer θM and K, during fallow. Macroporosity was determined based on the flow contribution between 0 and -5 cm water potentials (K0, K5, respectively), according to the Hagen-Poiseuille equation. The K0 values were statistically higher for CT than for NT and CP. The K5 values did not differ statistically among treatments. The mean K values varied between 0.20 and 3.70 cm/h. For CT, θM was significantly greater than for CP and NT, following the same trend as K0. No differences in θM were detected between CP and NT. With CT, the formation of water-conducting macropores with persistence until post-harvest was possible, while under CP preparation, the water-conducting macropores were not persistent. These results support the idea that tillage affects the soil water movement mainly by the resulting water-conducting macropores. Future studies on tillage effects on water movement should focus on macroporosity.

Impacts of deforestation on water balance components of a watershed on the Brazilian East Coast

The Brazilian East coast was intensely affected by deforestation, which drastically cut back the original biome. The possible impacts of this process on water resources are still unknown. The purpose of this study was an evaluation of the impacts of deforestation on the main water balance components of the Galo creek watershed, in the State of Espírito Santo, on the East coast of Brazil. Considering the real conditions of the watershed, the SWAT model was calibrated with data from 1997 to 2000 and validated for the period between 2001 and 2003. The calibration and validation processes were evaluated by the Nash-Sutcliffe efficiency coefficient and by the statistical parameters (determination coefficient, slope coefficient and F test) of the regression model adjusted for estimated and measured flow data. After calibration and validation of the model, new simulations were carried out for three different land use scenarios: a scenario in compliance with the law (C1), assuming the preservation of PPAs (permanent preservation areas); an optimistic scenario (C2), which considers the watershed to be almost entirely covered by native vegetation; and a pessimistic scenario (C3), in which the watershed would be almost entirely covered by pasture. The scenarios C1, C2 and C3 represent a soil cover of native forest of 76, 97 and 0 %, respectively. The results were compared with the simulation, considering the real scenario (C0) with 54 % forest cover. The Nash-Sutcliffe coefficients were 0.65 and 0.70 for calibration and validation, respectively, indicating satisfactory results in the flow simulation. A mean reduction of 10 % of the native forest cover would cause a mean annual increase of approximately 11.5 mm in total runoff at the watershed outlet. Reforestation would ensure minimum flows in the dry period and regulate the maximum flow of the main watercourse of the watershed.

Dietary effects on muscle fatty acid composition of finished heifers

The effects of diet on Longissimus muscle fatty acid composition was determined using 24 crossbred heifers of Simmental vs. Nelore and Limousin vs. Nelore. The experimental diets were: 1) corn and yeast (CY); 2) corn, cottonseed meal + meat and bones meal (CMB); 3) cassava hull and yeast (CHY); 4) cassava hull, cottonseed meal + meat and bones meal (CHMB). Feeding CHMB diets resulted in lower lipid and higher cholesterol contents (P<0.05) for both crosses. Most of the identified fatty acids were monounsaturated, and the highest percentage was found to oleic acid (C18:1w9), with values ranging from 32.54 to 46.42%. Among the saturated fatty acids the palmitic acid (C16:0) showed the highest percentage, with its contents ranging between 19.40 and 32.44%. The highest polyunsaturated/saturated fatty acid ratio was of 0.30, and the lowest was of 0.08. Feeding CY diets resulted in lower cholesterol and higher polyunsaturated fatty acid contents of the Longissimus muscle.

Crescimento e esporulação de isolados de Verticillium lecanii sob diferentes fatores ambientais

O objetivo deste trabalho foi avaliar o crescimento e a esporulação de dois isolados (JAB 02 e JAB 45) de Verticillium lecanii (Zimm.) Viégas, submetidos a diferentes valores de pH (4, 5, 6, 7, 8, 9) inicial do meio de cultivo sólido, de temperatura (19, 22, 25, 28 e 31ºC) e fotoperíodo (0, 12 e 24 horas). Após 20 dias de cultivo, não se observou efeito do pH inicial do meio no crescimento e esporulação dos isolados, exceto no pH 4,0 que reduziu o crescimento. A ausência de iluminação proporcionou a melhor condição para o crescimento de JAB 02 (32,70 mm) e JAB 45 (32,67 mm). Não houve efeito do fotoperíodo na produção de esporos. As melhores temperaturas para o crescimento de JAB 02 foram 19, 22 e 25ºC e para JAB 45 foram 19 e 25ºC. Este último isolado apresentou melhor esporulação a 19 e 22ºC, mas a temperatura não influenciou a esporulação de JAB 02. Não houve desenvolvimento dos isolados a 31ºC.

In vitro culture of Spondias mombin L. nodal segments

Spondias mombin L. shoot cultures were initiated from nodal explants taken from plants propagated by seeds. Explants coming from 4-6 months old plants, previously disinfected, were cultivated on WPM medium supplemented with a wide range of concentrations of BAP (0.0, 0.22, 0.44, 2.22 and 4.44 muM) and NAA (0.0, 0.27 and 2.70 muM). After four weeks, the responses obtained were axillary shoot and root formation. The first response were preferentially induced with the medium containing only BAP, regardless of the BAP concentration. The addition of NAA on medium reduced significantly axillary shoot formation and induced rhizogenesis. Roots were formed on nodal explant basis, preferentially on medium supplemented with 4.44 muM NAA. The medium supplemented with BAP reduced significantly root formation.

Monte Carlo derivation of filtered tungsten anode X-ray spectra for dose computation in digital mammography

Abstract Objective: Derive filtered tungsten X-ray spectra used in digital mammography systems by means of Monte Carlo simulations. Materials and Methods: Filtered spectra for rhodium filter were obtained for tube potentials between 26 and 32 kV. The half-value layer (HVL) of simulated filtered spectra were compared with those obtained experimentally with a solid state detector Unfors model 8202031-H Xi R/F & MAM Detector Platinum and 8201023-C Xi Base unit Platinum Plus w mAs in a Hologic Selenia Dimensions system using a direct radiography mode. Results: Calculated HVL values showed good agreement as compared with those obtained experimentally. The greatest relative difference between the Monte Carlo calculated HVL values and experimental HVL values was 4%. Conclusion: The results show that the filtered tungsten anode X-ray spectra and the EGSnrc Monte Carlo code can be used for mean glandular dose determination in mammography.