Seroprevalence of Toxoplasma gondii antibodies in humans from rural Western Amazon, Brazil

Antibodies to Toxoplasma gondii were assayed in sera of 266 humans from 71 farms located at Rond?nia State, Western Amazon, Brazil, by the modified agglutination test (MAT) and the indirect immunofluorescent antibody test (IFAT). Antibodies were found in 195 humans (73.3%), with MAT titers of 1:25 in 11, 1:50 in 11, 1:100 in 16, 1:200 in 27, 1:400 in 38, 1:800 in 37, 1:1,600 in 22, and 1:3,200 or higher in 33. From the 71 farms visited, 69 had seropositive humans. Prevalence of anti-T. gondii antibodies increased with age of the people (P < 0.05), and no difference was observed in the occurrence by gender (P > 0.05). A sanitary questionnaire was applied in each farm, and statistical association between the serologic status and several variables were analyzed. Home-grown vegetable consumption and origin of drinking water (well or river) were the independent variables that displayed significant association (P = 0.002 and 0.02, respectively). Higher values of occurrence were found in people with consumption of home-grown vegetables (76.1%) and people that drink well water (75.4%) compared with people that did not consume this type of food (61.9%) and drink river water (55.2%). By IFAT (> or = 1:16), 194 of 266 (73%) humans were seropositive and there was a good correlation between MAT and IFAT.     

Seroprevalence of Toxoplasma gondii antibodies in cats and pigs from rural Western Amazon, Brazil

Antibodies to Toxoplasma gondii were assayed in sera of 63 cats and 80 pigs from 71 farms located at Rond?nia State, Western Amazon, Brazil, by the modified agglutination test (MAT) and the indirect immunofluorescent antibody test (IFAT). Antibodies (MAT > or = 1: 25) were found in 55 of 63 cats (87.3%) with titers of 1:25 in 2, 1:50 in 2, 1:100 in 7, 1:200 in 1, 1:400 in 2, 1:800 in 9, 1:1,600 in 6, and 1:3,200 or higher in 26 cats. By IFAT, antibodies were found in 55 cats (87.3%) with titers of 1:25 in 2, 1:50 in 1, 1:100 in 4, 1:200 in 4, 1: 400 in 1, 1:800 in 13, 1:1,600 in 12, and 1:3,200 or higher in 18 cats. In pig sera, by MAT, antibodies were found in 30 of 80 pigs (37.5%) with titers of 1:25 in 2, 1:50 in 3, 1:100 in 2, 1:200 in 8, 1:400 in 3, 1:800 in 5, 1:1,600 in 3, and 1:3,200 or higher in 4 pigs. By using the IFAT (titers > or = 1:64), antibodies were found in 35 (43.7%) pigs. The ingestion of undercooked tissues of infected pigs can be a source of T. gondii infection for humans and cats. However, the high seroprevalence of T. gondii in cats from the Amazon seems most likely to be indicative of high contamination of the environment by oocysts.     

Full-genome sequences of hepatitis B virus subgenotype D3 isolates from the Brazilian Amazon Region

The Brazilian Amazon Region is a highly endemic area for hepatitis B virus (HBV). However, little is known regarding the genetic variability of the strains circulating in this geographical region. Here, we describe the first full-length genomes of HBV isolated in the Brazilian Amazon Region; these genomes are also the first complete HBV subgenotype D3 genomes reported for Brazil. The genomes of the five Brazilian isolates were all 3,182 base pairs in length and the isolates were classified as belonging to subgenotype D3, subtypes ayw2 (n = 3) and ayw3 (n = 2). Phylogenetic analysis suggested that the Brazilian sequences are not likely to be closely related to European D3 sequences. Such results will contribute to further epidemiological and evolutionary studies of HBV.     

Selenium Levels in the Whole Blood of Children and Teenagers from Two Riparian Communities at the Madeira River Basin in the Western Brazilian Amazon

Selenium (Se) is an essential micronutrient that exerts multiple functions in the organism, and both its deficiency and excess can cause health impairments. Thus, it is important to monitor its levels in the population, especially in vulnerable groups, such as children from the Brazilian Amazon region, where there is a lack of information in this regard. The aim of this research was to study Se levels in the whole blood of children and teenagers (5–16 years old) from two riparian communities at the Madeira River (Cuniã RESEX and Belmont). Se level variations related to the communities’ location, seasonality, diet, and body mass index (BMI) were assessed. Blood samples were collected in both communities for Se determinations, using ICP-MS and hemogram analyses, during May and September of 2011. Food frequency questionnaires were applied to assess consumption rates of specific food items. Non-parametric tests and linear multiple regressions were applied in the data analyses. Median Se levels were significantly higher during May (Cuniã RESEX 149 μg L^?1; Belmont 85 μg L^?1) compared to September (Cuniã RESEX 79 μg L^?1; Belmont 53 μg L^?1). No significant differences were found between the communities regarding BMI measurements and anemia prevalence. However, Se blood levels were significantly higher at the Cuniã RESEX compared to Belmont. In addition, the former showed higher fish and Brazil nut intakes, which may be the main Se sources for this community. These results contribute to a better understanding of Se reference levels for children and teenagers of Western Amazon riparian communities.     

Survey of Plasmodium spp. in Free-Ranging Neotropical Primates from the Brazilian Amazon Region impacted by Anthropogenic Actions

This study investigated Plasmodium spp. infection in free-ranging neotropical primates from Brazilian Amazon regions under the impact of major anthropogenic actions. Blood samples from 19 new world primates were collected and analyzed with microscopic and molecular procedures. The prevalence of Plasmodium infection was 21.0% (4/19) and PCR positive samples were identified as P. brasilianum. Considering the social-economic changes that the Amazon is facing, the prevalence of P. brasilianum infection highlights the necessity to closely monitor the movement of both human and non-human primate populations, in order to mitigate pathogen exposure and the introduction of new agents into previously naïve areas.     

Genotyping of two Neisseria gonorrhoeae fluroquinolone-resistant strains in the Brazilian Amazon Region

We report two ciprofloxacin and ofloxacin-resistant Neisseria gonorrhoeae strains that were isolated from the urethral discharge of male patients at the sexually transmitted diseases outpatient clinic of the Alfredo da Matta Foundation (Manaus, state of Amazonas, Brazil). The gonococci displayed minimal inhibitory concentrations (> 32.00 μg/mL) and three mutations in the quinolone resistance-determining region (S91F and D95G in GyrA and S87R in ParC). Both isolates were genotyped using N. gonorrhoeae multi-antigen sequence typing and the analysis showed that the ST225 which represented an emerging widespread multi-resistant clone that has also been associated with reduced susceptibility to ceftriaxone. We recommend continued surveillance of this pathogen to assess the efficacy of anti-gonococcal antibiotics in Brazil.     

Fire in the Brazilian Amazon

Regenerating forests have become a common land-cover type throughout the Brazilian Amazon. However, the potential for these systems to accumulate and store C and nutrients, and the fluxes resulting from them when they are cut, burned, and converted back to croplands and pastures have not been well quantified. In this study, we quantified pre- and post-fire pools of biomass, C, and nutrients, as well as the emissions of those elements, at a series of second- and third-growth forests located in the states of Pará and Rondônia, Brazil. Total aboveground biomass (TAGB) of second- and third-growth forests averaged 134 and 91 Mg ha^–1, respectively. Rates of aboveground biomass accumulation were rapid in these systems, but were not significantly different between second- and third-growth forests, ranging from 9 to 16 Mg ha^–1 year^–1. Residual pools of biomass originating from primary forest vegetation accounted for large portions of TAGB in both forest types and were primarily responsible for TAGB differences between the two forest types. In second-growth forests this pool (82 Mg ha^–1) represented 58% of TAGB, and in third-growth forests (40 Mg ha^–1) it represented 40% of TAGB. Amounts of TAGB consumed by burning of second- and third-growth forests averaged 70 and 53 Mg ha^–1, respectively. Aboveground pre-fire pools in second- and third-growth forests averaged 67 and 45 Mg C ha^–1, 821 and 707 kg N ha^–1, 441 and 341 kg P ha^–1, and 46 and 27 kg Ca ha^–1, respectively. While pre-fire pools of C, N, S and K were not significantly different between second- and third-growth forests, pools of both P and Ca were significantly higher in second-growth forests. This suggests that increasing land use has a negative impact on these elemental pools. Site losses of elements resulting from slashing and burning these sites were highly variable: losses of C ranged from 20 to 47 Mg ha^–1; N losses ranged from 306 to 709 kg ha^–1; Ca losses ranged from 10 to 145 kg ha^–1; and P losses ranged from 2 to 20 kg ha^–1. Elemental losses were controlled to a large extent by the relative distribution of elemental mass within biomass components of varying susceptibilities to combustion and the temperatures of volatilization of each element. Due to a relatively low temperature of volatilization and its concentration in highly combustible biomass pools, site losses of N averaged 70% of total pre-fire pools. In contrast, site losses of P and Ca resulting from burning were 33 and 20% of total pre-fire pools, respectively, as much of the mass of those elements was deposited on site as ash. Pre- and post-fire biomass and elemental pools of second- and third-growth forests, as well as the emissions from those systems, were intermediate between those of primary forests and pastures in the Brazilian Amazon. Overall, regenerating forests have the capacity to act as either large terrestrial sinks or sources of C and nutrients, depending on the course of land-use patterns within the Brazilian Amazon. Combining remote sensing techniques with field measures of aboveground C accumulation in regenerating forests and C fluxes from those forests when they are cut and burned, we estimate that during 1990–1991 roughly 104 Tg of C was accumulated by regenerating forests across the Brazilian Amazon. Further, we estimate that approximately 103 Tg of C was lost via the cutting and burning of regenerating forests across the Brazilian Amazon during this same period. Since average C accumulations (5.5 Mg ha^–1 year^–1) in regenerating forests were 19% of the C lost when such forests are cut and burned (29.3 Mg ha^–1), our results suggest that when less than 19% of the total area accounted for by secondary forests is cut and burned in a given year, those forests will be net accumulators of C during that year. Conversely, when more than 19% of regenerating forests are burned, those forests will be a net source of C to the atmosphere.     

Severity of Scorpion Stings in the Western Brazilian Amazon: A Case-Control Study

Background

Scorpion stings are a major public health problem in Brazil, with an increasing number of registered cases every year. Affecting mostly vulnerable populations, the phenomenon is not well described and is considered a neglected disease. In Brazil, the use of anti-venom formulations is provided free of charge. The associate scorpion sting case is subject to compulsory reporting. This paper describes the epidemiology and identifies factors associated with severity of scorpions stings in the state of Amazonas, in the Western Brazilian Amazon.

Methodology/Principal Findings

This study included all cases of scorpion stings in the state of Amazonas reported to the Brazilian Diseases Surveillance System from January 1, 2007 to December 31, 2014. A case-control study was conducted to identify factors associated with scorpions sting severity. A total of 2,120 cases were reported during this period. The mean incidence rate in the Amazonas was 7.6 per 100,000 inhabitants/year. Scorpion stings showed a large spatial distribution in the state and represent a potential occupational health problem for rural populations. There was a positive correlation between the absolute number of cases and the altimetric river levels in the Central (p<0.001; R_s = 0.479 linear) and Southwest (p = 0.032; linear R_s = 0.261) regions of the state. Cases were mostly classified as mild (68.6%), followed by moderate (26.8%), and severe (4.6%). The overall lethality rate was 0.3%. Lethality rate among children ≤10 years was 1.3%. Age <10 years [OR = 2.58 (95%CI = 1.47–4.55; p = 0.001)], stings occurring in the rural area [OR = 1.97 (95%CI = 1.18–3.29; p = 0.033) and in the South region of the state [OR = 1.85 (95%CI = 1.17–2.93; p = 0.008)] were independently associated with the risk of developing severity.

Conclusions/Significance

Scorpion stings show an extensive distribution in the Western Brazilian Amazon threatening especially rural populations, children ≤10 in particular. Thus, the mapping of scorpions fauna in different Amazon localities is essential and must be accompanied by the characterization of the main biological activities of the venoms. Urban and farming planning, in parallel with awareness of workers at risk for scorpion stings on the need for personal protective equipment use should be considered as public policies for preventing scorpionism.     

15N natural abundance in forest and pasture soils of the Brazilian Amazon Basin

The natural abundance of ¹⁵N was examined in soil profiles from forests and pastures of the Brazilian Amazon Basin to compare tropical forests on a variety of soil types and to investigate changes in the sources of nitrogen to soils following deforestation for cattle ranching. Six sites in the state of Rondônia, two sites in Pará and one in Amazonas were studied. All sites except one were chronosequences and contained native forest and one or more pastures ranging from 2 to 27 years old. Forest soil ¹⁵N values to a depth of 1 m ranged from 8 to 23 and were higher than values typically found in temperate forests. A general pattern of increasing ¹⁵N values with depth near the soil surface was broadly similar to patterns in other forests but a decrease in ¹⁵N values in many forest profiles between 20 and 40 cm suggests that illuviation of ¹⁵N-depleted nitrate may influence total soil ¹⁵N values in deeper soil where total N concentrations are low. In four chronosequences in Rondônia, the ¹⁵N values of surface soil from pastures were lower than in the original forest and ¹⁵N values were increasingly depleted in older pastures. Inputs of atmospheric N by dinitrogen fixation could be an important N source in these pastures. Other pastures in Amazonas and Pará and Rondônia showed no consistent change from forest values. The extent of fractionation that leads to ¹⁵N enrichment in soils was broadly similar over a wide range of soil textures and indicated that similar processes control N fractionation and loss under tropical forest over a broad geographic region. Forest ¹⁵N profiles were consistent with conceptual models that explain enrichment of soil ¹⁵N values by selective loss of ¹⁴N during nitrification and denitrification.     

JC polyomavirus infection in candidates for kidney transplantation living in the Brazilian Amazon Region

This study evaluated the relative occurrences of BK virus (BKV) and JC virus (JCV) infections in patients with chronic kidney disease (CKD). Urine samples were analysed from CKD patients and from 99 patients without CKD as a control. A total of 100 urine samples were analysed from the experimental (CKD patients) group and 99 from the control group. Following DNA extraction, polymerase chain reaction (PCR) was used to amplify a 173 bp region of the gene encoding the T antigen of the BKV and JCV. JCV and BKV infections were differentiated based on the enzymatic digestion of the amplified products using BamHI endonuclease. The results indicated that none of the patients in either group was infected with the BKV, whereas 11.1% (11/99) of the control group subjects and 4% (4/100) of the kidney patients were infected with the JCV. High levels of urea in the excreted urine, low urinary cellularity, reduced bladder washout and a delay in analysing the samples may have contributed to the low prevalence of infection. The results indicate that there is a need to increase the sensitivity of assays used to detect viruses in patients with CDK, especially given that polyomavirus infections, especially BKV, can lead to a loss of kidney function following transplantation.     

Asplundia altiscandens sp. nov. (Cyclanthaceae) from the Brazilian Amazon

A new species, Asplundia altiscandens E. S. Leal from the Brazilian Amazon, is described and illustrated. It belongs to subgenus Asplundia and is related to A. ferruginea and A. nonoensis. Diagnostic characters, a distribution map and taxonomic comments are provided.     

A new Culicoides (Diptera: Ceratopogonidae) of the reticulatus species group from Brazilian Amazon Region

A new species of the Culicoides reticulatus species group, Culicoides kuripako Felippe-Bauer, is described and illustrated based on females and male specimens from the states of Amazonas and Pará, Brazil. The new species is compared with its similar sympatric congener, Culicoides paucienfuscatus Barbosa.     

Natural abundance of 15N in soils along forest-to-pasture chronosequences in the western Brazilian Amazon Basin

We examined the natural abundance of ¹⁵N in soil profiles along two chronosequences in the western Brazilian Amazon Basin state of Rondônia, to investigate possible mechanisms for changes to soil nitrogen sources and transformations that occur as a result of land use. One chronosequence consisted of forest and 3-, 5- and 20-year-old pasture, the other of forest and 8- and 20-year-old pasture. The ¹⁵N values of surface soil and soil to 1 m depth in the native forest ranged from 9.8 to 13.6 and were higher than reported for temperate forest soils. Fractionation associated with nitrification and denitrification and selective losses of ¹⁵N-depleted nitrate, could potentially result in a strong enrichment of nitrogen in soil organic matter over the time scale of soil development in highly weathered tropical soils. Pasture surface soils were 1–3, depleted in ¹⁵N compared with forest soils. Lower ¹⁵N values in 20-year-old pastures is consistent with greater cumulative inputs of ¹⁵N-depleted atmospheric-derived nitrogen, fixed by free-living bacteria associated with planted pasture grasses in older pastures, or differential plant utilization of soil inorganic N pools with different ¹⁵N values. The pattern of ¹⁵N values following conversion of forest to agricultural use differs from the pattern in the temperate zone, where pasture or cultivated soils are typically more enriched in ¹⁵N than the forest soils from which they were derived.     

Comparison of the ant communities of annually inundated and terra firme forests at Trombetas in the Brazilian Amazon

Summary The composition of the ant community was assessed along standardized 100 m transects in annually flooded Varzea forest and in terra firme forests on sandy soil (Flanco forest) and on claytopped mesas (Planalto forest). Standardized samples were taken by unit-time hand collecting (day and night times), sweeping, beating, baiting and by Winkler sacks. A total of 156 species, representing 49 genera were found, of which 98, 88 and 55 were respectively found in the Planalto, Flanco and Varzea forests. Species lists are presented and the ant community composition and species richness are compared between the three forests. By considering the nesting and foraging habits of the various species, the differences in overall community composition are related to the forest type and susceptibility to inundation of the three forests which were surveyed.The data confirm the view that tropical rain forests support an extremely diverse ant fauna and comparisons with other forested areas suggest that ant species richness declines in subtropical and temperate rain forests. Although alpha diversity is high, species turnover between forests is lower than expected, suggesting that ant species richness in this forested region is not as great as is implied in some published estimates of global arthropod diversity.     

The responses of leaf litter ant communities to wildfires in the Brazilian Amazon: a multi-region assessment

Fire is frequently used as tool for land management in the Amazon, but often escapes into surrounding forests, with potentially severe impacts for forest biodiversity. We investigated the effects of single wildfires on ant communities in four geographically distinct regions of the Brazilian Amazon (Roraima, Pará, Acre and Mato Grosso) where forests had burned between 8 months and 10 years before our sampling. We established 7–12 transects, 500 m each, in burned and unburned forests in each region to investigate the effects of fire on forest structure and leaf litter ant communities, which were sampled using Winkler sacks. Fire effects on forest structure were more drastic in the most recently burned forests in Acre and Mato Grosso, while the impacts of older burns in Roraima and Pará were more subtle. Ant species richness was not different between burnt and unburned areas, but community composition differed between burned and control forests in all regions except Mato Grosso. At the species level, indicator species analysis showed that a limited number of species were significant indicators of unburned control forests in all regions, except Acre. Forests structure variables and leaf litter volume were all important in shaping ant communities, but their relative importance varied between regions. Our results indicate that burned forest have different ant species communities from unburned forests, and those differences are still apparent 10 years after the disturbance, highlighting the importance of effective policies for fire management in Amazon.     

Biomass of primary and secondary vegetation in Rondônia, Western Brazilian Amazon

Biomass estimates of primary and different ages of secondary vegetation are reported for a tropical forest region in Rondônia, Western Brazilian Amazon. The estimates are based on published allometric equations, and on vegetation composition and allometric data collected in areas of primary forest and secondary vegetation of ages 2, 3, 5, 9, 11, 16 and 18 years. Primary forest biomass estimates varied from 290 to 495 t ha^–1. Secondary vegetation biomass estimates accounted for 40–60% of the primary forest biomass after 18 years of abandonment. Secondary growth rates in lightly used areas are estimated to have varied from 6.6 to 8.7 t ha^–1 y^–1 between the third and the eighteenth years after abandonment. CO₂ sequestration by regrowing vegetation is discussed for two scenarios of land abandonment.     

Comparative cytogenetics of three economically important Piper L. species from the Brazilian Amazon

The species Piper hispidinervum, Piper aduncum, and Piper affinis hispidinervum have essential oils with high levels of safrole, dillapiole, and sarisan, respectively. Safrole is important for pharmaceutical and chemical industries, while dillapiole and sarisan are promising compounds to control insects and fungi. These species are very similar morphologically and their taxonomy is controversial. Divergent hypotheses consider P. aduncum and P. hispidinervum either as a single species or as distinct taxa, while P. affinis hispidinervum is inferred to be a natural hybrid or a chemotype of P. hispidinervum. Delimiting the taxonomic boundaries would be helpful for germplasm conservation and breeding programs. This study aimed to undertake a detailed analysis of P. aduncum, P. hispidinervum, and P. affinis hispidinervum karyotype and rDNA sites. Genomic in situ hybridization (GISH) was used to establish genomic homology among species and to test the natural hybridization hypothesis for origin of P. affinis hispidinervum. Karyotype traits were similar for all three species: 2n = 26 small chromosomes, predominantly metacentric. All three species exhibited CMA+ bands on the secondary constriction of chromosome pair 4. A size-heteromorphic 35S rDNA site was co-localized with the CMA+ band. A 5S rDNA site was located in the proximal region of chromosome pair 7. The patterns of genomic hybridization revealed that the repetitive DNA fraction of the species is highly similar in terms of proportion of genome, sequence type, and distribution. Our findings did not allow us to differentiate the three species and point to the importance of deeper genomic studies to elucidate the taxonomic controversy.

     

Spatial Analysis Spotlighting Early Childhood Leprosy Transmission in a Hyperendemic Municipality of the Brazilian Amazon Region

Background

More than 200,000 new cases of leprosy were reported by 105 countries in 2011. The disease is a public health problem in Brazil, particularly within high-burden pockets in the Amazon region where leprosy is hyperendemic among children.

Methodology

We applied geographic information systems and spatial analysis to determine the spatio-temporal pattern of leprosy cases in a hyperendemic municipality of the Brazilian Amazon region (Castanhal). Moreover, we performed active surveillance to collect clinical, epidemiological and serological data of the household contacts of people affected by leprosy and school children in the general population. The occurrence of subclinical infection and overt disease among the evaluated individuals was correlated with the spatio-temporal pattern of leprosy.

Principal Findings

The pattern of leprosy cases showed significant spatio-temporal heterogeneity (p<0.01). Considering 499 mapped cases, we found spatial clusters of high and low detection rates and spatial autocorrelation of individual cases at fine spatio-temporal scales. The relative risk of contracting leprosy in one specific cluster with a high detection rate is almost four times the risk in the areas of low detection rate (RR = 3.86; 95% CI = 2.26–6.59; p<0.0001). Eight new cases were detected among 302 evaluated household contacts: two living in areas of clusters of high detection rate and six in hyperendemic census tracts. Of 188 examined students, 134 (71.3%) lived in hyperendemic areas, 120 (63.8%) were dwelling less than 100 meters of at least one reported leprosy case, 125 (66.5%) showed immunological evidence (positive anti-PGL-I IgM titer) of subclinical infection, and 9 (4.8%) were diagnosed with leprosy (8 within 200 meters of a case living in the same area).

Conclusions/Significance

Spatial analysis provided a better understanding of the high rate of early childhood leprosy transmission in this region. These findings can be applied to guide leprosy control programs to target intervention to high risk areas.