A FRET-Based Real-Time PCR Assay to Identify the Main Causal Agents of New World Tegumentary Leishmaniasis

In South America, various species of Leishmania are endemic and cause New World tegumentary leishmaniasis (NWTL). The correct identification of these species is critical for adequate clinical management and surveillance activities. We developed a real-time polymerase chain reaction (PCR) assay and evaluated its diagnostic performance using 64 archived parasite isolates and 192 prospectively identified samples collected from individuals with suspected leishmaniasis enrolled at two reference clinics in Lima, Peru. The real-time PCR assay was able to detect a single parasite and provided unambiguous melting peaks for five Leishmania species of the Viannia subgenus that are highly prevalent in South America: L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, L. (V.) peruviana and L. (V.) lainsoni. Using kinetoplastid DNA-based PCR as a gold standard, the real-time PCR had sensitivity and specificity values of 92% and 77%, respectively, which were significantly higher than those of conventional tests such as microscopy, culture and the leishmanin skin test (LST). In addition, the real-time PCR identified 147 different clinical samples at the species level, providing an overall agreement of 100% when compared to multilocus sequence typing (MLST) data performed on a subset of these samples. Furthermore, the real-time PCR was three times faster and five times less expensive when compared to PCR - MLST for species identification from clinical specimens. In summary, this new assay represents a cost-effective and reliable alternative for the identification of the main species causing NWTL in South America.     

Root colonization and spore abundance of arbuscular mycorrhizal fungi in distinct successional stages from an Atlantic rainforest biome in southern Brazil

The influence of plant functional groups and moderate seasonality on arbuscular mycorrhizal (AM) fungal status (root colonization and spore density) was investigated during 13 consecutive months in a chronosequence of succession in southern Brazil, consisting of grassland field, scrub vegetation, secondary forest and mature forest, in a region of transition from tropical to subtropical zones. AM root colonization and spore density decreased with advancing succession and were highest in early successional sites with grassland and scrub vegetation, intermediary in the secondary forest and lowest in the mature forest. They were little influenced by soil properties, but were sufficiently influenced by the fine root nutrient status and fine root traits among different functional plant groups. AM root colonization and spore density were higher during the favourable plant growth season (spring and summer) than during the less favourable plant growth season (autumn and winter). Spore density displayed significant seasonal variation at all sites, whilst root colonization displayed significant seasonal variation in grassland, scrub and secondary forest, but not in mature forest. The data suggest that (1) different plant functional groups display different relationships with AM fungi, influencing their abundance differentially; (2) plant species from early successional phases are more susceptible to AM root colonization and maintain higher AM sporulation than late successional species; (3) fine root traits and nutrient status influence these AM fungal attributes; and (4) higher AM spore production and root colonization is associated with the season of higher light incidence and temperature, abundant water in soil and higher plant metabolic activity.     

Wind matters: Asymmetric distribution of aphids on host plants can be explained by stems functioning as windbreaks

Understanding how abiotic factors influence organisms at present is the necessary first step to predict how species assemblages could be affected by climate change in the future. We examined how wind, a poorly studied abiotic factor, affects the distribution and abundance of two aphid species, Uroleucon aeneum and Brachycaudus cardui (hereafter black and green aphids, respectively), that live on the thistle Carduus thoermeri (Asteraceae) in a windy region of Patagonia, Argentina. First, considering the prevailing wind direction, we described the distribution of both aphid species around plant stems. Then, we performed a bi‐factorial experiment in which we cut stems with aphids to manipulate their position respect to wind (exposed/unexposed) and to control wind incidence (protected/unprotected). Finally, using the species most affected by wind, we examined possible mechanisms through which wind could affect aphids. Both aphid species were less abundant on the side of the stem exposed to wind respect to the unexposed side; and this pattern was stronger for the black aphid. When black aphids were positioned exposed to wind and without protection, their proportion changed towards the unexposed side of the stem; while green aphids showed a weaker response to wind. Laboratory experiments demonstrated that wind triggered both the detachment of black aphids and their movement towards the unexposed side of the stem. Our results showed that wind can explain the asymmetric distribution of aphids around plants and that stems can act as windbreaks. In a less windy future scenario, aphids could expand their foraging area, reaching higher infestation rates, which could affect their role in structuring ant assemblages and their status as crop pests. This work highlights the importance of testing the effects of less studied abiotic factors to fully understand how climate change could impact on the abundance and distribution of animals in the future.     

Diarrhea outbreak during U.S. military training in El Salvador

Infectious diarrhea remains a major risk to deployed military units worldwide in addition to their impact on travelers and populations living in the developing world. This report describes an outbreak of diarrheal illness in the U.S. military’s 130^th Maneuver Enhancement Brigade deployed in San Vicente, El Salvador during a training and humanitarian assistance mission. An outbreak investigation team from U.S. Naval Medical Research Unit – Six conducted an epidemiologic survey and environmental assessment, patient interviews, and collected stool samples for analysis in an at risk population of 287 personnel from May 31^st to June 3^rd, 2011. Personnel (n = 241) completed an epidemiological survey (87% response rate) and 67 (27%) reported diarrhea and/or vomiting during the past two weeks. The median duration of illness was reported to be 3 days (IQR 2–4 days) and abdominal pain was reported among 30 (49%) individuals. Presentation to the medical aid station was sought by (62%) individuals and 9 (15%) had to stop or significantly reduce work for at least one day. Microscopy and PCR analysis of 14 stool samples collected from previously symptomatic patients, Shigella (7), Cryptosporidium (5), and Cyclospora (4) were the most prevalent pathogens detected. Consumption of food from on-base local vendors (RR = 4.01, 95% CI = 1.53–10.5, p-value <0.001) and arriving on base within the past two weeks (RR = 2.79, 95% confidence [CI] = 1.35–5.76, p-value = 0.001) were associated with increased risk of developing diarrheal disease. The risk of infectious diarrhea is great among reserve military personnel during two week training exercises. The consumption of local food, prepared without proper monitoring, is a risk factor for deployed personnel developing diarrheal illness. Additional information is needed to better understand disease risks to personnel conducting humanitarian assistance activities in the Latin America Region.     

Investment in Fine Roots and Arbuscular Mycorrhizal Fungi Decrease During Succession in Three Brazilian Ecosystems

The functional groups of plants that characterize different phases of succession are expected to show differences in root distribution, fine‐root traits and degrees of association with arbuscular mycorrhizal (AM) fungi. The relationship involving fine‐root traits and AM fungi that regulate the nutrient acquisition potential among different plant functional groups are still not well understood. We assessed fine‐root morphology, AM fungal variables and soil fertility in grassland, secondary forest and mature forest in Atlantic, Araucaria and Pantanal ecosystems in Brazil. Soil cores were collected at 0–10 and 10–20 cm depths. Fine roots were extracted from soil by sieving and root morphological traits and AM colonization were determined. The AM spores were extracted from soil and counted. In all ecosystems, soil fertility, fine‐root mass and root diameter increased with the succession, while root length, specific root length, root‐hair length, root‐hair incidence, AM colonization and AM spore density decreased. These results suggest that plant species from early stages of tropical succession with inherent rapid growth invest in fine roots and maintain a high degree of AM colonization in order to increase the capacity for nutrient acquisition. Conversely, fine root morphological characteristics and low degree of AM colonization exhibited by plants of the later stages of succession lead toward a low nutrient uptake capacity that combine with their typical low growth rates. Abstract in Portuguese is available at http://www.blackwell‐synergy.com/loi/btp .     

Essential but invisible: non-apparent but widespread ant nests favour soil nutrients and plant growth in semi-arid areas

1. Ants provide multiple ecosystem services, including nutrient cycling. Although most studies on nests effects on soil fertility and plant performance include species with large nests, species with less apparent nests may have a relevant effect, especially if they are widespread, abundant, and/or inhabit nutrient-poor soils. 2. We studied the effects of nests of three abundant and widespread ant species in the Patagonian steppe (the generalist Dorymyrmex tener, and the seed harvesters Pogonomyrmex carbonarius and Pheidole spininodis) on soil properties, plant growth of two native species, and seedling recruitment. Our main hypothesis was that, despite their non-apparent nests, these species have a positive effect on soils and enhance plant growth. 3. Nest soils showed higher soil conductivity, %K and %Mg than non-nest soils. In a greenhouse experiment, individuals of the biennial forb Oenothera odorata grew bigger in nest soils of P. carbonarius and D. tener than in non-nest soils. Individuals of the perennial tussock grass Pappostipa speciosa grew taller and had more tillers in nest versus non-nest soils. Seedling abundance and richness were the highest in P. carbonarius nest soils and the general trends were similar to those observed in the plant growth experiment. 4. Our results show that ant species with non-apparent nests in an arid area enhance soil properties, favouring plant growth (nests of P. carbonarius and D. tener) and seedling abundance (nests of P. carbonarius). Due to their high abundance and widespread distribution, these two species could have a relevant role in ecosystem recovery after disturbance.     

Native Rodent Species Are Unlikely Sources of Infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios,Peru

An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru''s Amazon region, the department of Madre de Dios (MDD) rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia) braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vector-borne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V.) braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail) were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%), Hylaeamys perenensis (15.7%), and Proechimys spp. (12%). All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V.) braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V.) braziliensis transmission to human inhabitants in this highly prevalent region.     

Functional traits reveal environmental constraints on amphibian community assembly in a subtropical dry forest

The relationships between functional traits and environmental gradients are useful to identify different community assembly processes. In this work, we used an approach based on functional traits to analyse if changes in hydroperiod and tree covers of ponds are relevant for local amphibian community assembly processes. Ephemeral ponds with low vegetation cover are expected to impose constraints on different species with particular trait combinations and, therefore, to exhibit communities with lower functional diversity than more stable ponds with greater tree cover. Sampling was conducted in 39 temporary ponds located along vegetation and hydroperiod gradients in the most arid portion of the Chaco ecoregion. Seven functional traits were measured in each species present in the regional pool. Associations between these traits and environmental gradients were detected using multivariate ordination techniques and permutation test (RLQ and fourth‐corner analyses respectively). Functional diversity indices were then calculated and related to variations in the environmental gradients. The results obtained allowed us to identify different sets of traits associated with hydroperiod and tree cover, suggesting that these environmental variables are relevant for structuring amphibian communities according to interspecific variations in functional traits from both, larval and adult stages. Contrary to our expectations, communities associated with more stable ponds and with greater tree cover exhibited lower functional diversity than expected by chance (and were the ponds with highest species richness). This result indicates that the reduction in relative importance of environmental restrictions imposed by a very short hydroperiod and the lack of tree cover, favours different species of the regional pool that are similar in several functional traits. Accordingly, communities associated with stable ponds with high tree cover exhibited high functional redundancy.     

Syndromic Management and STI Control in Urban Peru

Background

Syndromic management is an inexpensive and effective method for the treatment of symptomatic sexually transmitted infections (STIs), but its effectiveness as a method of STI control in at-risk populations is questionable. We sought to determine the potential utility of syndromic management as a public health strategy to control STI transmission in high-risk populations in urban Peru.

Methodology

We surveyed 3,285 at-risk men and women from three Peruvian cities from 2003–05. Participants were asked about the presence of genital ulcers, discharge, or dysuria in the preceding six months. Participants reporting symptoms were asked about subsequent health-seeking and partner notification behavior. Urine and vaginal swab samples were tested for Neisseria gonorrhoeae and Chlamydia trachomatis by nucleic acid testing. Serum was tested for syphilis and Herpes Simplex Virus-Type 2 antibodies.

Findings

Recent urogenital discharge or dysuria was reported by 42.1% of participants with gonorrhea or chlamydia versus 28.3% of participants without infection. Genital ulceration was reported by 6.2% of participants with, and 7.4% of participants without, recent syphilis. Many participants reporting symptoms continued sexual activity while symptomatic, and approximately half of all symptomatic participants sought treatment. The positive and negative predictive values of urogenital discharge or genital ulcer disease in detecting STIs that are common in the study population were 14.4% and 81.5% for chlamydia in women and 8.3% and 89.5% for syphilis among gay-identified men.

Conclusions

In our study, STIs among high-risk men and women in urban Peru were frequently asymptomatic and symptomatic participants often remained sexually active without seeking treatment. Additional research is needed to assess the costs and benefits of targeted, laboratory-based STI screening as part of a comprehensive STI control program in developing countries.     

Pay it forward: refuse dump from leaf‐cutting ants promotes caterpillar digestive performance by increasing plant nitrogen content

1. The activity of soil‐disturbing animals that increase soil nutrients can affect the carbon : nitrogen (C : N) ratio of plants, which, in turn, may determine the transfer of energy and nutrients through higher trophic levels. However, the strength and sign of this indirect effect depend on whether enhanced nutrient substrates increase plant foliar nutrients and/or plant defensive traits. 2. We investigated how the nutrient‐rich refuse dumps of the leaf‐cutting ant Acromyrmex lobicornis, as a result of their direct effects on thistles, indirectly impact the growth rate and digestive performance of a generalist chewing herbivore. We also included the application of commercial fertilisers to test whether the enhanced soil nutrients comprises the mechanism behind the impact of refuse dumps on the upward cascade effects. 3. Thistles growing on nutrient‐rich substrates (i.e. ant refuse dumps and fertilised soil) had more and larger leaves, up to 80% lower C : N ratios, and higher physical defences compared to plants growing on steppe soil. Caterpillars showed an enhanced digestive performance and growth rate when feed on nutrient‐rich plants and were able to adjust the C : N ratio of their excretion to regulate the relative acquisition of nutrients. 4. The positive effect of nutrient‐rich substrates on caterpillar feeding efficiency suggest that the enhanced nutritional quality of the thistles could compensate for the negative effects of the increased physical defences. The results of the present study indicate how organisms that increase soil nutrient availability may diminish the stoichiometric constraints at the base of food chains, enhancing the development and growth rate of herbivores and, thus, indirectly mediating plant–herbivore interactions.