Cyanobacterial bloom monitoring and assessment in Latin America.

Cyanobacterial blooms have serious adverse effects on human and environmental health. In Latin America, one of the main world's freshwater reserves, information on this phenomenon remains sparse. To assess the current situation, we gathered reports of cyanobacterial blooms and associated cyanotoxins in freshwater bodies from South America and the Caribbean (Latitude 22° N to 45° S) and compiled the regulation and monitoring procedures implemented in each country. As the operational definition of what is a cyanobacterial bloom remains controversial, we also analyzed the criteria used to determine the phenomena in the region. From 2000 to 2019, blooms were reported in 295 water bodies distributed in 14 countries, including shallow and deep lakes, reservoirs, and rivers. Cyanotoxins were found in nine countries and high concentrations of microcystins were reported in all types of water bodies. Blooms were defined according to different, and sometimes arbitrary criteria including qualitative (changes in water color, scum presence), quantitative (abundance), or both. We found 13 different cell abundance thresholds defining bloom events, from 2 × 103 to 1 × 107 cells mL-1. The use of different criteria hampers the estimation of bloom occurrence, and consequently the associated risks and economic impacts. The large differences between countries in terms of number of studies, monitoring efforts, public access to the data and regulations regarding cyanobacteria and cyanotoxins highlights the need to rethink cyanobacterial bloom monitoring, seeking common criteria. General policies leading to solid frameworks based on defined criteria are needed to improve the assessment of cyanobacterial blooms in Latin America. This review represents a starting point toward common approaches for cyanobacterial monitoring and risk assessment, needed to improve regional environmental policies.

Nutrients and not temperature are the key drivers for cyanobacterial biomass in the Americas.

Cyanobacterial blooms imperil the use of freshwater around the globe and present challenges for water management. Studies have suggested that blooms are trigged by high temperatures and nutrient concentrations. While the roles of nitrogen and phosphorus have long been debated, cyanobacterial dominance in phytoplankton has widely been associated with climate warming. However, studies at large geographical scales, covering diverse climate regions and lake depths, are still needed to clarify the drivers of cyanobacterial success. Here, we analyzed data from 464 lakes covering a 14,000 km north-south gradient in the Americas and three lake depth categories. We show that there were no clear trends in cyanobacterial biomass (as biovolume) along latitude or climate gradients, with the exception of lower biomass in polar climates. Phosphorus was the primary resource explaining cyanobacterial biomass in the Americas, while nitrogen was also significant but particularly relevant in very shallow lakes (< 3 m depth). Despite the assessed climatic gradient water temperature was only weakly related to cyanobacterial biomass, suggesting it is overemphasized in current discussions. Depth was critical for predicting cyanobacterial biomass, and shallow lakes proved more vulnerable to eutrophication. Among other variables analyzed, only pH was significantly related to cyanobacteria biomass, likely due to a biologically mediated positive feedback under high nutrient conditions. Solutions toward managing harmful cyanobacteria should thus consider lake morphometric characteristics and emphasize nutrient control, independently of temperature gradients, since local factors are more critical - and more amenable to controls - than global external forces.

Complete de novo assembly of Wolbachia endosymbiont of Diaphorina citri Kuwayama (Hemiptera: Liviidae) using long-read genome sequencing.

Wolbachia, a gram-negative [Formula: see text]-proteobacterium, is an endosymbiont found in some arthropods and nematodes. Diaphorina citri Kuwayama, the vector of 'Candidatus Liberibacter asiaticus' (CLas), are naturally infected with a strain of Wolbachia (wDi), which has been shown to colocalize with the bacteria pathogens CLas, the pathogen associated with huanglongbing (HLB) disease of citrus. The relationship between wDi and CLas is poorly understood in part because the complete genome of wDi has not been available. Using high-quality long-read PacBio circular consensus sequences, we present the largest complete circular wDi genome among supergroup-B members. The assembled circular chromosome is 1.52 megabases with 95.7% genome completeness with contamination of 1.45%, as assessed by checkM. We identified Insertion Sequences (ISs) and prophage genes scattered throughout the genomes. The proteins were annotated using Pfam, eggNOG, and COG that assigned unique domains and functions. The wDi genome was compared with previously sequenced Wolbachia genomes using pangenome and phylogenetic analyses. The availability of a complete circular chromosome of wDi will facilitate understanding of its role within the insect vector, which may assist in developing tools for disease management. This information also provides a baseline for understanding phylogenetic relationships among Wolbachia of other insect vectors.

Nuisance phytoplankton transport is enhanced by high flow in the main river for drinking water in Uruguay.

Eutrophication, climate change, and river flow fragmentation are the main cause of nuisance algal blooms worldwide. This study evaluated the conditions that trigger the growth and occurrence of nuisance phytoplankton in the Santa Lucía River, a subtropical floodplain lotic system that supplies drinking water to 60% of the population of Uruguay. The main variables that explained phytoplankton biovolume were extracted from generalized linear models (GLM). The potential impact of nuisance organism advection on water utility was estimated by the phytoplankton biovolume transport (BVTR, m3 day-1), an indicator of biomass load. Santa Lucía River had a wide flow range (0.7×105-1438×105 m3 day-1) and eutrophic conditions (median, TP: 0.139 mg L-1; TN: 0.589 mg L-1). GLMs indicated that phytoplankton biomass increased with temperature and soluble reactive phosphorus. Contrary to expectations, the presence of cyanobacteria was positively associated with periods of high flow that result in high cyanobacterial biovolume transport, with a probability of 3.35 times higher when flow increased by one standard deviation. The cyanobacterial biovolume transported (max: 9.5 m3 day-1) suggests that biomass was subsidized by allochthonous inocula. Biovolume from other nuisance groups (diatoms, cryptophytes, and euglenophytes) was positively associated with low-flow conditions and high nutrient concentrations in the main river channel, thereby indicating that these conditions boost eukaryote blooms. The evaluation of BVTR allows a better understanding of the dynamics of fluvial phytoplankton and can help to anticipate scenarios of nuisance species transport.

Metagenomic analysis of Raphidiopsisraciborskii microbiome: beyond the individual.

Raphidiopsisraciborskii is a toxic, invasive bacteria with a defined biogeographic pattern attributed to the generation of ecotypes subjected to local environmental filters and to phenotypic plasticity. The interactions taking place between the cyanobacterium and the other bacteria inhabiting the external polysaccharide-rich matrix surrounding the cells, or phycosphere, may be ecotype-specific and would have different influence on the carbon and nutrient cycling in the ecosystem. Here, we describe the bacterial community or microbiome (assessed by 16S rRNA metagenomics) associated to two R.raciborskii strains that have been described as different ecotypes: the saxitoxin-producer MVCC19 and the non-toxic LB2897. Our results showed that both ecotypes share 50% of their microbiomes and differ in their dominant taxa. The taxon having the highest abundance in the microbiome of MVCC19 was Neorhizobium (22.5% relative abundance), while the dominant taxon in LB2897 was the Planctomycetes SM1A02 (26.2% relative abundance). These groups exhibit different metabolic capabilities regarding nitrogen acquisition (symbiotic nitrogen-fixing in Neorhizobium vs. anammox in SM1A02), suggesting the existence of ecotype-specific microbiomes that play a relevant role in cyanobacterial niche-adaptation. In addition, as saxitoxin and analogues are nitrogen-rich (7 atoms per molecule), we hypothesise that saxitoxin-producing R.raciborskii benefits from external sources of nitrogen provided by the microbiome bacteria. Based on these findings, we propose that the mechanisms involved in the assembly of the cyanobacterial microbiome community are ecotype-dependent.

Antibacterial FANA oligonucleotides as a novel approach for managing the Huanglongbing pathosystem.

Candidatus Liberibacter asiaticus (CLas), a bacterium transmitted by the Asian citrus psyllid, Diaphorina citri, is the causal agent of citrus greening disease, or Huanglongbng (HLB). Currently, vector population suppression with insecticides and tree removal are the most effective strategies for managing the HLB pathosystem. In this study, we assessed the bactericidal capabilities of 2'-deoxy-2'-fluoro-D-arabinonucleic acid antisense oligonucleotides (FANA ASO) both in vitro and in vivo by (1) confirming their capacity to penetrate insect cells, (2) silencing bacterial essential genes, and (3) quantifying reductions in bacterial titer and D. citri transmission. We confirmed that FANA ASO are able to penetrate insect cells without the use of a delivery agent. Expression of an essential gene in the D. citri endosymbiont, Wolbachia (wDi), significantly decreased by 30% following incubation with a wDi-specific FANA ASO. Viability of isolated wDi cells also decreased in response to the FANA ASO treatment. Delivery of a CLas-specific FANA ASO to infected adult D. citri in feeding assays resulted in significant silencing of a CLas essential gene. CLas relative density and transmission were significantly lower among D. citri fed FANA ASO in diet compared to untreated insects. Root infusions of a CLas-specific FANA ASO into infected Citrus trees significantly reduced CLas titer during a 30-day trial. Our results suggest that FANA ASO targeting insect-transmitted plant bacteria or insect endosymbionts may be useful tool for integrated management of agricultural pathogens.

Severidad de hipomineralización incisivo molar (HIM) y su relación con caries dental en niños / Severity of incisor molar hypomineralization (IMH) and its relationship with dental caries in children

Artículo original Original articleRevista Metro CienciaSeveridad de hipomineralización incisivo molar (HIM) y su relación con caries dental en niñosSeverity of incisor molar hypomineralization (IMH) and its relationship with dental caries in childrenSylvia Gabriela Pineda Bonilla1, Alejandra Cabrera Arias2Odontóloga de la Universidad Católica del Ecuador, Quito, Ecuador1Docente de la Universidad Católica del Ecuador, Quito, Ecuador2Recibido: 28/09/2020 Aceptado:05/10/2020 Públicado:29/10/2020Editorial: Hospital Metropolitano ISSN (impreso) 1390-2989 - ISSN (electrónico) 2737-6303Edición: Vol. 28 Nº 4 (2020) octubre - diciembreDOI: https://doi.org/10.47464/MetroCiencia/vol28/4/2020/42-51URL: https://revistametrociencia.com.ec/index.php/revista/article/view/95Pág: 42-51RESUMENLa Hipomineralización Incisivo Molar (HIM) es una alteración cualitativa en la formación de la estructura del diente que puede producir la pérdida de grandes áreas de la estructura dental. La prevalencia de Hipomineralización Incisivo Molar varía ampliamente en el mundo y han sido pocos los estudios epidemio-lógicos realizados en Latinoamérica y específicamente en Ecuador. Objetivo: Determinar la prevalencia y severidad de (HIM) y su asociación con caries dental en niños y niñas de 8 a 10 años de escuelas primarias públicas del Barrio Las Casas. Materiales y métodos: Corresponde a un estudio, transversal en el cual se evaluó a 366 escolares de 8 a 10 años de edad que asistían a las escuelas públicas del barrio Las Casas, que tuvieran todos los primeros molares e incisivos permanentes completamente erupcionados. Las variables fueron edad, género, escolaridad del representante, ocupación del representante, fre-cuencia de cepillado; las variables clínicas que se evaluaron fueron HIM mediante los criterios descritos por Mathu-Muju y Wright; y caries dental, utilizando el índice International Caries Detection and Assessment System (ICDAS II). Resultados: La prevalencia de hipomineralización incisivo molar en la muestra total fue de 21.3%, al asociarla con la caries dental se detectó que el 2.2% (n=8) de escolares que no presentaba caries presentó HIM, a diferencia del 9.3% (n=34) de escolares que presentaron caries con cavidades detectables y el 9.8% (n=36) de niños con lesiones incipientes tuvieron HIM. Conclusión: La presencia de HIM fue alta y si estuvo asociada a caries dental, los escolares con HIM presentaron más riesgo de desarrollar caries.

ABSTRACT Molar Incisor Hypomineralization (MIH) is a qualitative alteration in the formation of tooth structure that can cause the loss of large areas of tooth structure. The prevalence of Molar Incisor Hypomineralization varies widely in the world and there have been few epidemiological studies conducted in Latin America and specifically in Ecuador. Objective: To determine the prevalence and severity of (MIH) and its association with dental caries in children from 8 to 10 years old of public primary schools in Las Casas neighborhood. Material and methods: Corresponds to a cross-sectional study in which 366 schoolchildren between 8 and 10 years old were evaluated who attended to the public schools in Las Casas neighborhood, which had all the first molars and permanent incisors completely erupted. The variables were age, gender, education of the representative, occupation of the representative, frequency of brushing; the clinical variables that were evaluated were HIM using the criteria described by Mathu-Muju & Wright; and dental caries, using the International Caries Detection and Assessment System index (ICDAS II). Results: The prevalence of molar incisor hypomineralization in the total sample was 21.3% when it was associating with dental caries it was detected that 2.2% (n = 8) of schoolchildren without caries presented MIH, a difference of 9.3% (n = 34) of schoolchildren who presented caries with detectable cavities and 9.8% (n = 36) of children with incipient lesions had MIH. Conclusion: The presence of MIH was high and was associated with dental caries. Schoolchildren with MIH presented a higher risk of developing caries.

Near-Complete Genome Sequences of a Wolbachia Strain Isolated from Diaphorina citri Kuwayama (Hemiptera: Liviidae).

Wolbachia strains are one of three endosymbionts associated with the insect vector of "Candidatus Liberibacter asiaticus," Diaphorina citri Kuwayama (Hemiptera: Liviidae). We report three near-complete genome sequences of samples of Wolbachia from D. citri (wDi), with sizes of 1,518,595, 1,542,468, and 1,538,523 bp.

Predicting cyanobacterial biovolume from water temperature and conductivity using a Bayesian compound Poisson-Gamma model.

Eutrophication and climate change scenarios engender the need to develop good predictive models for harmful cyanobacterial blooms (CyanoHABs). Nevertheless, modeling cyanobacterial biomass is a challenging task due to strongly skewed distributions that include many absences as well as extreme values (dense blooms). Most modeling approaches alter the natural distribution of the data by splitting them into zeros (absences) and positive values, assuming that different processes underlie these two components. Our objectives were (1) to develop a probabilistic model relating cyanobacterial biovolume to environmental variables in the Río de la Plata Estuary (35°S, 56°W, n = 205 observations) considering all biovolume values (zeros and positive biomass) as part of the same process; and (2) to use the model to predict cyanobacterial biovolume under different risk level scenarios using water temperature and conductivity as explanatory variables. We developed a compound Poisson-Gamma (CPG) regression model, an approach that has not previously been used for modeling phytoplankton biovolume, within a Bayesian hierarchical framework. Posterior predictive checks showed that the fitted model had a good overall fit to the observed cyanobacterial biovolume and to more specific features of the data, such as the proportion of samples crossing three threshold risk levels (0.2, 1 and 2 mm³ L-1) at different water temperatures and conductivities. The CPG model highlights the strong control of cyanobacterial biovolume by nonlinear and interactive effects of water temperature and conductivity. The highest probability of crossing the three biovolume levels occurred at 22.2 °C and at the lowest observed conductivity (∼0.1 mS cm-1). Cross-validation of the fitted model using out-of-sample observations (n = 72) showed the model's potential to be used in situ, as it enabled prediction of cyanobacterial biomass based on two readily measured variables (temperature and conductivity), making it an interesting tool for early alert systems and management strategies. Furthermore, this novel application demonstrates the potential of the Bayesian CPG approach for predicting cyanobacterial dynamics in response to environmental change.

Biogeography of the cyanobacterium Raphidiopsis (Cylindrospermopsis) raciborskii: Integrating genomics, phylogenetic and toxicity data.

Raphidiopsis (Cylindrospermopsis) raciborskii, a globally distributed bloom-forming cyanobacterium, produces either the cytotoxin cylindrospermopsin (CYL) in Oceania, Asia and Europe or the neurotoxin saxitoxin (STX) and analogues (paralytic shellfish poison, PSP) in South America (encoded by sxt genetic cluster) and none of them in Africa. Nevertheless, this particular geographic pattern is usually overlooked in current hypotheses about the species dispersal routes. Here, we combined genomics, phylogenetic analyses, toxicity data and a literature survey to unveil the evolutionary history and spread of the species. Phylogenies based on 354 orthologous genes from all the available genomes and ribosomal ITS sequences of the taxon showed two well-defined clades: the American, having the PSP producers; and the Oceania/Europe/Asia, including the CYL producers. We propose central Africa as the original dispersion center (non-toxic populations), reaching North Africa and North America (in former Laurasia continent). The ability to produce CYL probably took place in populations that advanced to sub-Saharan Africa and then to Oceania and South America. According to the genomic context of the sxt cluster found in PSP-producer strains, this trait was acquired once by horizontal transfer in South America, where the ability to produce CYL was lost.

Identifying populations most susceptible to get benefit from broadening the scope for prevention of cervical cancer: Example from Uruguay.

OBJECTIVES: To identify factors associated with high-risk human papillomavirus (HPV) infection and high grade squamous intraepithelial lesion (HSIL) among a high-risk group of HPV-unvaccinated women in Montevideo. METHODS: Participants completed a questionnaire on socio-demographics, sexual behavior and gynecological history and received a gynecological examination. HPV DNA was detected by PCR using MY09/11 primers. Logistic regression analyses were performed to identify factors associated with high-risk HPV infection and HSIL. RESULTS: A total of 469 women with HPV DNA and cytological results completed the questionnaire. Among women older than 30 years, those with high number of sexual partners and regular housing conditions were more likely to be positive for high-risk HPV infection (adjusted OR: 2.94, 95%CI: 1.01-8.51 and 2.68, 95%CI: 1.01-7.21, respectively). A marginally non-statistically significant association between getting a HSIL and having a high number of sexual partners was also observed (adjusted OR: 3.22, 95%CI: 0.97-10.75). CONCLUSIONS: In an era of development of new strategies for accelerating the reduction of cervical cancer incidence and mortality, our results may contribute to identify populations most susceptible to get benefit from broadening the scope for prevention of cervical cancer and could be used with other triage strategies.

Analysis of human papillomavirus 16 E6, E7 genes and Long Control Region in cervical samples from Uruguayan women.

OBJECTIVE: This study aims to investigate the HPV16 variant distribution by sequence analyses of E6, E7 oncogenes and the Long Control Region (LCR), from cervical cells collected from Uruguayan women, and to reconstruct the phylogenetic relationships among variants. METHODS: Forty-seven HPV16 variants, obtained from women with HSIL, LSIL, ASCUS and NILM cytological classes were analyzed for LCR and 12 were further studied for E6 and E7. Detailed sequence comparison, genetic heterogeneity analyses and phylogenetic reconstruction were performed. RESULTS: A high variability was observed among LCR sequences, which were distributed in 18 different variants. E6 and E7 sequences exhibited novel non-synonymous substitutions. Uruguayan sequences mainly belonged to the European lineage, and only 5 sequences clustered in non-European branches; 3 of them in the Asian-American and North-American linage and 2 in an African branch. Additionally, 6 new variants from European and African clusters were identified. CONCLUSIONS: HPV16 isolates mainly belonged to the European lineage, though strains from African and Asian-American lineages were also identified. Herein is reported for the first time the distribution and molecular characterization of HPV16 variants from Uruguay, providing novel insights on the molecular epidemiology of this infectious disease in the South America. SYNOPSIS: A high variability among HPV 16 isolates mainly belonged to European lineage, provides an extensive sequence dataset from a country with high burden of cervical cancer.

Temperature and precipitation shape the distribution of harmful cyanobacteria in subtropical lotic and lentic ecosystems.

Cyanobacterial blooms are expected to become more frequent in freshwaters globally due to eutrophication and climate change effects. However, our knowledge about cyanobacterial biogeography in the subtropics, particularly in lotic ecosystems, is still very limited and the relationship of blooms to temperature and precipitation remains unclear. We took advantage of a comprehensive database of field data compiled over several years (1997 to 2015) to compare cyanobacteria biomass and distribution between lentic and lotic subtropical freshwaters (36 ecosystems, 30°-35°S) and to investigate the role of water temperature and precipitation as significant predictors in eutrophic ecosystems. A filamentous Nostocales, Dolichospermum (Anabaena), was the most widely distributed and frequent genus in the region of the study, followed by the colonial Microcystis, supporting observations of a global latitudinal pattern. Similar total cyanobacteria biovolumes (TCB) were found in lentic and lotic ecosystems, but the proportion of Dolichospermum was higher in lotic ecosystems. Using generalized linear models (GLMs), we found that temperature and rainfall explained 27% of the variation in TCB in lotic ecosystems, while temperature explained 19 and 28% of Dolichospermum and Microcystis biovolume, respectively. In lentic ecosystems, accumulated rainfall explained 34% of the variation of Microcystis biovolume while temperature explained 64%. Our results imply that the increase in extreme meteorological events and temperature predicted by climate models will promote increasingly severe cyanobacterial blooms in eutrophic subtropical freshwaters. Our analysis provides new information about the occurrence of bloom-forming cyanobacteria for southeastern South America and thus fills an important knowledge gap for subtropical freshwaters.

Correction: A Naturally-Derived Compound Schisandrin B Enhanced Light Sensation in the pde6c Zebrafish Model of Retinal Degeneration.

[This corrects the article DOI: 10.1371/journal.pone.0149663.].

A Naturally-Derived Compound Schisandrin B Enhanced Light Sensation in the pde6c Zebrafish Model of Retinal Degeneration.

Retinal degeneration is often progressive. This feature has provided a therapeutic window for intervention that may extend functional vision in patients. Even though this approach is feasible, few promising drug candidates are available. The scarcity of new drugs has motivated research to discover novel compounds through different sources. One such example is Schisandrin B (SchB), an active component isolated from the five-flavor fruit (Fructus Schisandrae) that is postulated in traditional Chinese medicines to exert prophylactic visual benefit. This SchB benefit was investigated in this study in pde6cw59, a zebrafish retinal-degeneration model. In this model, the pde6c gene (phosphodiesterase 6C, cGMP-specific, cone, alpha prime) carried a mutation which caused cone degeneration. This altered the local environment and caused the bystander rods to degenerate too. To test SchB on the pde6cw59 mutants, a treatment concentration was first determined that would not cause morphological defects, and would initiate known physiological response. Then, the mutants were treated with the optimized SchB concentration before the appearance of retinal degeneration at 3 days postfertilization (dpf). The light sensation of animals was evaluated at 6 dpf by the visual motor response (VMR), a visual startle that could be initiated by drastic light onset and offset. The results show that the VMR of pde6cw59 mutants towards light onset was enhanced by the SchB treatment, and that the initial phase of the enhancement was primarily mediated through the mutants' eyes. Further immunostaining analysis indicates that the treatment specifically reduced the size of the abnormally large rods. These observations implicate an interesting hypothesis: that the morphologically-improved rods drive the observed VMR enhancement. Together, these investigations have identified a possible visual benefit of SchB on retinal degeneration, a benefit that can potentially be further developed to extend functional vision in patients.

Influence of nitrogen availability on the expression of genes involved in the biosynthesis of saxitoxin and analogs in Cylindrospermopsis raciborskii.

The development of cyanobacterial blooms in inland aquatic ecosystems is greatly promoted by nutrient availability, especially nitrogen and phosphorous. When blooms are dominated by toxigenic species the harmful effects of nutrient loading becomes particularly relevant. Among toxic species, Cylindrospermopsis raciborskii found in South American ecosystems is characterized by the production of saxitoxin and analogs (Paralytic Shellfish Poisoning, PSP), for which the factors that trigger their production have not been elucidated. In this study, the effect of nitrate availability on the relative transcript abundance of two genes (sxtU and sxtI), both involved in different steps of PSP biosynthetic pathway, was addressed in C. raciborskii MVCC19 by qPCR. The relative transcript abundance of both genes significantly increased from the beginning to the end of growth, independent of nitrate availability in the culture medium. Differences between the genes in terms of the levels of relative expression were also found, implying that during growth in nitrate-rich or nitrate-deprived conditions C. raciborskii MVCC19 has the ability to produce different kind of PSP molecules. The presence of nifH transcripts in the nitrogen-depleted treatment confirmed that in the absence of nitrate C. raciborskii fixed atmospheric N2. Moreover, after transferring filaments to nitrate-rich conditions the synthesis of nifH mRNA continued for few hours, suggesting that cell adjustments enabling the utilization of soluble nitrogen sources are not immediate. Our results show that biosynthesis of saxitoxin and analogs in C. raciborskii is not related to nitrate availability, but rather is linked to cyanobacteria growth rate.

Combining immunolabeling and catalyzed reporter deposition to detect intracellular saxitoxin in a cyanobacterium.

We combined the use of polyclonal antibodies against saxitoxin with catalyzed reporter deposition to detect production of saxitoxin by the cyanobacterium Cylindrospermopsis raciborskii. The procedure is simple, allows detection of intracellular saxitoxin in cyanobacteria filaments by confocal laser microscopy and is a promising tool to study toxin production and metabolism.

The chromatin remodeler chd5 is necessary for proper head development during embryogenesis of Danio rerio.

The chromatin remodeler CHD5 plays a critical role in tumor suppression and neurogenesis in mammals. CHD5 contributes to gene expression during neurogenesis, but there is still much to learn regarding how this class of remodelers contributes to differentiation and development. CHD5 remodelers are vertebrate-specific, raising the prospect that CHD5 plays one or more conserved roles in this phylum. Expression of chd5 in adult fish closely mirrors expression of CHD5 in adult mammals. Knockdown of Chd5 during embryogenesis suggests new roles for CHD5 remodelers based on resulting defects in craniofacial development including reduced head and eye size as well as reduced cartilage formation in the head. In addition, knockdown of Chd5 results in altered expression of neural markers in the developing brain and eye as well as a profound defect in differentiation of dopaminergic amacrine cells. Recombinant zebrafish Chd5 protein exhibits nucleosome remodeling activity in vitro, suggesting that it is the loss of this activity that contributes to the observed phenotypes. Our studies indicate that zebrafish is an appropriate model for functional characterization of CHD5 remodelers in vertebrates and highlight the potential of this model for generating novel insights into the role of this vital class of remodelers.

Application of ancient DNA to the reconstruction of past microbial assemblages and for the detection of toxic cyanobacteria in subtropical freshwater ecosystems.

Ancient DNA (aDNA) analysis of lake sediments is a promising tool for detecting shifts in past microbial assemblages in response to changing environmental conditions. We examined sediment core samples from subtropical, freshwater Laguna Blanca (Uruguay), which has been severely affected by cultural eutrophication since 1960 and where cyanobacterial blooms, particularly those of the saxitoxin-producer Cylindrospermopsis raciborskii, have been reported since the 1990s. Samples corresponding to ~1846, 1852, 2000 and 2007 AD were selected to perform denaturing gradient gel electrophoresis (DGGE) analysis of the 16S-23S rRNA intergenic transcribed spacer (ribosomal ITS) to compare their prokaryotic assemblage composition. Each stratum showed different ITS patterns, but the composition of 21st century samples was clearly different than those of mid-19th century. This compositional change was correlated with shifts in sediment organic matter and chlorophyll a content, which were significantly higher in recent samples. The presence of saxitoxin-producing cyanobacteria was addressed by quantitative real-time PCR of the sxtU gene involved in toxin biosynthesis. This gene was present only in recent samples, for which clone libraries and ITS sequencing indicated the presence of Cyanobacteria. Phylogenetic analyses identified C. raciborskii only in the 2000 sample, shortly after several years when blooms were recorded in the lake. These data suggest the utility of aDNA for the reconstruction of microbial assemblage shifts in subtropical lakes, at least on centennial scales. The application of aDNA analysis to genes involved in cyanotoxin synthesis extends the applicability of molecular techniques in palaeolimnological studies to include key microbial community characteristics of great scientific and social interest.

p35 promotes the differentiation of amacrine cell subtype in the zebrafish retina under the regulation of egr1.

BACKGROUND: Early growth response 1 (egr1) is a transcription factor (TF) for controlling the differentiation of Parvalbumin (Parv) -expressing amacrine cells (ACs) in zebrafish. However, the downstream factors of this process have not been identified. The purpose of this study was to investigate the role of p35, a neuronal-specific activator of cyclin-dependent kinase 5 (Cdk5) and a known in vitro target of egr1, in the differentiation of these ACs. RESULTS: In the p35-knockdown retinas, Parv+ but not islet1+ ACs were specifically reduced. This phenotype was highly similar to that in the Egr1-knockdown retinas. Furthermore, p35 expression was reduced in the Egr1-knockdown retinas, particularly in the AC region; while egr1 was only modestly reduced in this region in the p35-knockdown retinas. CONCLUSIONS: p35 likely acts downstream of egr1 to control the differentiation of Parv+ ACs.