Virus-Bacterium Coupling Driven by both Turbidity and Hydrodynamics in an Amazonian Floodplain Lake

The importance of viruses in aquatic ecosystem functioning has been widely described. However, few studies have examined tropical aquatic ecosystems. Here, we evaluated for the first time viruses and their relationship with other planktonic communities in an Amazonian freshwater ecosystem. Coupling between viruses and bacteria was studied, focusing both on hydrologic dynamics and anthropogenic forced turbidity in the system (Lake Batata). Samples were taken during four hydrologic seasons at both natural and impacted sites to count virus-like particles (VLP) and bacteria. In parallel, virus-infected bacteria were identified and quantified by transmission electron microscopy (TEM). Viral abundance ranged from 0.5 × 10⁷ ± 0.2 × 10⁷ VLP ml⁻¹ (high-water season, impacted site) to 1.7 × 10⁷ ± 0.4 × 10⁷ VLP ml⁻¹ (low-water season, natural site). These data were strongly correlated with the bacterial abundance (r² = 0.84; P < 0.05), which ranged from 1.0 × 10⁶ ± 0.5 × 10⁶ cells ml⁻¹ (high water, impacted site) to 3.4 × 10⁶ ± 0.7 × 10⁶ cells ml⁻¹ (low water, natural site). Moreover, the viral abundance was weakly correlated with chlorophyll a, suggesting that most viruses were bacteriophages. TEM quantitative analyses revealed that the frequency of visibly infected cells was 20%, with 10 ± 3 phages per cell section. In general, we found a low virus-bacterium ratio (<7). Both the close coupling between the viral and bacterial abundances and the low virus-bacterium ratio suggest that viral abundance tends to be driven by the reduction of hosts for viral infection. Our results demonstrate that viruses are controlled by biological substrates, whereas in addition to grazing, bacteria are regulated by physical processes caused by turbidity, which affect underwater light distribution and dissolved organic carbon availability.Viruses are the most abundant and dynamic components of the aquatic microbial community (6, 31, 32). Viruses influence many biogeochemical and ecological processes, including nutrient cycling, system respiration, particle-size distribution, bacterial and algal biodiversity, species distribution, algal blooms, and genetic transfer between microorganisms (21, 49). In addition, viruses play a major role in aquatic microbial food webs by releasing carbon trapped in host cells to the dissolved organic carbon (DOC) pool and ultimately back to the bacterial community (11, 21). The action of viruses is an important mechanism of bacterial regulation in aquatic ecosystems, acting directly on bacterial populations and indirectly on bacterial diversity by decreasing the density of dominant bacterial species (31). Studies based on viral decay rates and electron microscopy analyses have shown that viruses can cause up to 40% of bacterial mortality and more than 10% of phytoplankton mortality in aquatic systems (11, 22, 48, 50, 54, 55). It also has been suggested that viral lysis and protistan grazing cause similar bacterial mortality in aquatic ecosystems (22, 40).Several environmental factors, including solar radiation and temperature, can influence viral abundance. Exposure to solar radiation decreases viral abundance in aquatic ecosystems, while low temperatures decrease their virulence (33, 56). However, the majority of the studies on virus ecology have been performed in temperate or polar regions, where seasonal changes in solar radiation and water temperature are more pronounced (26, 30, 32). Viral abundances have been little investigated in tropical aquatic ecosystems (6, 39) and particularly in the Amazonian region, where the abundance and activity of aquatic viruses have not been studied.The greatest watershed in the world is located in the Amazonian region. It is composed of clear-water, black-water, and turbid freshwater ecosystems, which are seasonally influenced by the flood pulse. The hydrologic pulse is characterized by a pronounced change in water level, defining the flood seasons. Nutrient sources and stocks and species dynamics vary according to the water level (25). During the high-water season (flood season), the tight connection between terrestrial and aquatic environments results in an increase in allochthonous DOC input and the dilution of inorganic nutrients and organisms. During the low-water season, there is an increase in nutrient concentrations, organism abundances, and the importance of autochthonous DOC. These seasonal changes differently impact ecosystem functions and aquatic community dynamics (5, 9, 18). For instance, bacterioplankton abundance is less changeable than phytoplankton abundance throughout the hydrological cycle due to the alternative sources of DOC (allochthonous in the high-water period and autochthonous in the low-water period) for bacterial communities (5, 24).Lake Batata is a clear-water Amazonian floodplain lake located in the watershed of the Trombetas River, a tributary of the Amazon River. As a clear-water Amazonian ecosystem, it contains low concentrations of suspended particles and inorganic nutrients (47). Lake Batata is distinct because it was impacted by bauxite tailings for 10 years (1979 to 1989), affecting 30% of the lake''s area. The tailings caused a huge increase in turbidity; large amounts of tailings settled on the sediment surface and often are resuspended by physical mixing or biotic movements (28). The presence of tailings resulted in a clear spatial variation in the lake, forming impacted and natural sites. Furthermore, tailing particles can directly act as a substrate for attaching bacteria and also can adsorb DOC (5).Previous studies on bacterio-, phyto-, and zooplankton communities have shown that flood pulse acts as the primary driver of plankton community structure in Lake Batata (5, 9, 24). Bauxite tailings also affect microbial processes in impacted sites, such as bacterial growth and production (5), photosynthesis rates and primary production (43), or the availability of food for zooplankton (8). However, there still is no evidence indicating a complementary (flood pulse and forced turbidity) effect among these factors in any microbial community. Based on published data, we assume that (i) bacterioplankton abundance is less changeable through the hydrologic cycle than phytoplankton abundance (5, 24), and (ii) tailing particles can act as a substrate for attaching bacteria and also can adsorb organic matter, which is controlled by the flood pulse (5). Therefore, we hypothesized that the relationship between viruses and bacteria in Lake Batata is modulated by a synergistic effect between the hydrological cycle and turbidity.     

Influence of carrion smell and rebaiting time on the efficiency of pitfall traps to dung beetle sampling

Dung beetles (Coleoptera: Scarabaeidae: Scarabaeinae) are very useful insects, as they improve the chemo-physical properties of soil, clean pastures from dung pads, and help control symbovine flies associated with bovine cattle. Their importance makes it fundamental to sample and survey them adequately. The objectives of the present study were to determine the influence of decaying insects trapped in pitfalls on the attractiveness of Moura pig Sus scrofa L. (Suidae) and collared peccary Tayassu tajacu (L.) (Tayassuidae) dung used as baits to lure dung beetles, and to establish how long these baits remain attractive to dung beetles when used in these traps. Some dung beetle species seemed to be able to discriminate against foul smell from decaying insects within the first 24 h, hence decreasing trap efficiency. This was more evident in peccary dung-baited traps, which proved to be the least attractive bait. Attractiveness lasted only 24 h for peccary dung, after which it became unattractive, whereas the pig dung bait was highly attractive for 48 h, after which its attractiveness diminished but was not completely lost.     

Complete Genome Sequences of Corynebacterium pseudotuberculosis Strains 3/99-5 and 42/02-A, Isolated from Sheep in Scotland and Australia, Respectively

Here, we report the whole-genome sequences of two ovine-pathogenic Corynebacterium pseudotuberculosis isolates: strain 3/99-5, which represents the first C. pseudotuberculosis genome originating from the United Kingdom, and 42/02-A, the second from Australia. These genome sequences will contribute to the objective of determining the global pan-genome of this bacterium.     

Ethnopharmacological survey among migrants living in the Southeast Atlantic Forest of Diadema, São Paulo, Brazil

Background

Understanding how people of diverse cultural backgrounds have traditionally used plants and animals as medicinal substances during displacements is one of the most important objectives of ethnopharmacological studies. An ethnopharmacological survey conducted among migrants living in the Southeast Atlantic Forest remnants (Diadema, São Paulo, Brazil) is presented herein.

Methods

Ethnographical methods were used to select and interview the migrants, and botanical and zoological techniques were employed to collect the indicated resources.

Results

We interviewed five migrants who described knowledge on 12 animals and 85 plants. Only 78 plants were present in Diadema, they belong to 37 taxonomic families; 68 were used exclusively for medicinal purposes, whereas 10 were reported to be toxic and/or presented some restriction of use. These taxa were grouped into 12 therapeutic categories (e.g., gastrointestinal disturbances, inflammatory processes or respiratory problems) based on the 41 individual complaints cited by the migrants. While the twelve animal species were used by the migrants to treat nine complaints; these were divided into six categories, the largest of which related to respiratory problems. None of the animal species and only 57 of the 78 plant species analysed in the present study were previously reported in the pharmacological literature; the popular knowledge concurred with academic findings for 30 of the plants. The seven plants [Impatiens hawkeri W. Bull., Artemisia canphorata Vill., Equisetum arvensis L., Senna pendula (Humb. & Bonpl. ex Willd.) H.S. Irwin & Barneby, Zea mays L., Fevillea passiflora Vell. and Croton fuscescens Spreng)] and the two animals (Atta sexdens and Periplaneta americana) that showed maintenance of use among migrants during their displacement in Brazilian territory, have not been studied by pharmacologists yet.

Conclusions

Thus, they should be highlighted and focused in further pharmacology and phytochemical studies, since the persistence of their uses can be indicative of bioactive potentials.     

Dissolved methane concentrations and fluxes to the atmosphere from a tropical floodplain lake

Large uncertainties in estimates of methane (CH₄) emissions from tropical inland waters reflect the paucity of information at appropriate temporal and spatial scales. CH₄ concentrations, diffusive and ebullitive fluxes, and environmental parameters in contrasting aquatic habitats of Lake Janauacá, an Amazon floodplain lake, measured for two years revealed patterns in temporal and spatial variability related to different aquatic habitats and environmental conditions. CH₄ concentrations ranged from below detection to 96 µM, CH₄ diffusive fluxes from below detection to 2342 µmol m⁻² h⁻¹, and CH₄ ebullitive fluxes from 0 to 190 mmol m⁻² d⁻¹. Vegetated aquatic habitats had higher surface CH₄ concentrations than open water habitats, and no significant differences in diffusive CH₄ fluxes, likely due to higher k values measured in open water habitats. CH₄ emissions were enhanced after a prolonged low water period, when the exposed sediments were colonized by herbaceous plants that decomposed after water levels rose, possibly fueling CH₄ production. Statistical models indicated the importance of variables related to CH₄ production (temperature, dissolved organic carbon) and consumption (dissolved nitrogen, oxygenated water column), as well as maximum depth, in controlling surface water CH₄ concentrations.

     

Helmet‐shaped Body of Entodiniomorphid Ciliates (Ciliophora,Entodiniomorphida), a Synapomorphy or a Homoplasy?

Triadinium was created to include Triadinium caudatum. Further, four other species were included, T. minimum, T. galea, T. elongatum, and T. magnum, all sharing a characteristic helmet‐shaped body. Wolska and Grain argued that the inclusion of T. minimum and T. galea into Triadinium was done based on superficial morphological aspects, and established two new genera to accommodate these species: Circodinium and Gassovskiella. Although the phylogenetic relationships within Entodiniomorphida have been investigated by multiple authors, none of them discussed the evolutionary relationship of helmet‐shaped entodiniomorphids. We performed molecular phylogenetics and revisited old literature digging for morphological data to explain our results. According to our analyses, the helmet‐shaped body is homoplastic and may have evolved from at least three different entodiniomorphid ancestors. Circodinium minimum is phylogenetically related to members of Blepharocorythidae, T. caudatum emerged within Spirodiniidae and G. galea within Polydiniellidae. This phylogenetic hypothesis is partially supported by information on infraciliature and ultrastructure of C. minimum and T. caudatum. However, such morphological information is not available for polydiniellids. In order to shed some light into the evolution of the helmet‐shaped ciliates, future works should focus to collect information on the infraciliature and the ultrastructure of Polydiniella mysorea and of other Triadinium species.     

Porcine Adipose Tissue-Derived Mesenchymal Stem Cells Retain Their Proliferative Characteristics,Senescence, Karyotype and Plasticity after Long-Term Cryopreservation

We and others have provided evidence that adipose tissue-derived mesenchymal stem cells (ASCs) can mitigate rat cardiac functional deterioration after myocardial ischemia, even though the mechanism of action or the relevance of these findings to human conditions remains elusive. In this regard, the porcine model is a key translational step, because it displays heart anatomic-physiological features that are similar to those found in the human heart. Towards this end, we wanted to establish the cultural characteristics of porcine ASCs (pASCs) with or without long-term cryostorage, considering that allogeneic transplantation may also be a future option. Compared to fresh pASCs, thawed cells displayed 90–95% viability and no changes in morphological characteristics or in the expression of surface markers (being pASCs characterized by positive markers CD29⁺; CD90⁺; CD44⁺; CD140b⁺; CD105⁺; and negative markers CD31⁻; CD34⁻; CD45⁻ and SLA-DR⁻; n = 3). Mean population doubling time was also comparable (64.26±15.11 hours to thawed cells vs. 62.74±18.07 hours to fresh cells) and cumulative population doubling increased constantly until Passage 10 (P10) in the entire cell population, with a small and gradual increase in senescence (P5, 3.25%±0.26 vs. 3.47%±0.32 and P10, 9.6%±0.29 vs. 10.67%±1.25, thawed vs. fresh; SA-β-Gal staining). Chromosomal aberrations were not observed. In addition, under both conditions pASCs responded to adipogenic and osteogenic chemical cues in vitro. In conclusion, we have demonstrated the growth characteristics, senescence, and the capacity of pASCs to respond to chemical cues in vitro and have provided evidence that these properties are not influenced by cryostorage in 10% DMSO solution.     

Fecal Microbial Diversity in Pre-Weaned Dairy Calves as Described by Pyrosequencing of Metagenomic 16S rDNA. Associations of Faecalibacterium Species with Health and Growth

In this study, we use barcoded pyrosequencing of the 16S rRNA gene to characterize the fecal microbiota of neonatal calves and identify possible relationships of certain microbiota profiles with health and weight gain. Fecal samples were obtained weekly from 61 calves from birth until weaning (seventh week of the calves'' life). Firmicutes was the most prevalent phylum, with a prevalence ranging from 63.84% to 81.90%, followed by Bacteroidetes (8.36% to 23.93%), Proteobacteria (3.72% to 9.75%), Fusobacteria (0.76% to 5.67%), and Actinobacteria (1.02% to 2.35%). Chao1 index gradually increased from the first to the seventh postnatal week. Chao1 index was lower during the third, fourth, and fifth week of life in calves that suffered from pneumonia and were treated with antibiotics. Diarrhea incidence during the first four weeks of the calves'' life was also associated with a reduction of microbial diversity during the third week of life. Increased fecal microbial diversity after the second week of life was associated with higher weight gain. Using discriminant analysis we were able to show differences in the microbiota profiles between different weeks of life, between high and low weight gain groups of calves, and between calves affected and not affected with diarrhea during the first four weeks life. The prevalence of Faecalibacterium spp. in the first week of life was associated with weight gain and the incidence of diarrhea, with higher prevalence being associated with higher weight gain and less diarrhea. Representative sequences from Faecalibacterium spp. were closely affiliated to Faecalibacterium prausnitzii. Results presented here provide new information regarding the intestinal microbiota of neonatal calves and its association with health and growth. Fecal microbial diversity was associated with calf age, disease status and growth rates. Results suggesting a possible beneficial effect of Faecalibacterium spp. on health and growth are promising.     

In Vitro–In Vivo Correlation of Efavirenz Tablets Using GastroPlus®

The aim of the present work was to use GastroPlus™ software for the prediction of pharmacokinetic profiles and in vitro–in vivo correlation (IVIVC) as tools to optimize the development of new generic medications. GastroPlus™ was used to simulate the gastrointestinal compartment and was based on the advanced compartmental absorption and transit model. Powder dissolution and efavirenz tablet dissolution studies were carried out to generate data from which correlation was established. The simulated plasma profile, based on the physicochemical properties of efavirenz, was almost identical to that observed in vivo for biobatches A and B. A level A IVIVC was established for the dissolution method obtained for the generic candidate using the Wagner–Nelson (r² = 0.85) and for Loo–Riegelman models (r² = 0.92). The percentage of fraction absorbed indicated that 0.5% sodium lauryl sulfate may be considered a biorelevant dissolution medium for efavirenz tablets. The simulation of gastrointestinal bioavailability and IVIVC obtained from immediate-release tablet formulations suggests that GastroPlus™ is a valuable in silico method for IVIVC and for studies directed at developing formulations of class II drugs.KEY WORDS: bioavailability, computational simulation, efavirenz, GastroPlus™, in vivo–in vitro correlation