Toxicity of selected pharmaceuticals and their mixtures to the aquatic indicators Daphnia magna and Aliivibrio fischeri.

Despite the benefits derived from the use of pharmaceuticals, these compounds are currently considered contaminants of emerging concern because of their presence and persistence in the environment. This study aimed to determine the toxicity of 27 pharmaceuticals and the interaction effects of binary mixtures of selected compounds towards two model organisms: the microcrustacean Daphnia magna and the bacterium Aliivibrio fischeri (Microtox test). Six compounds, namely polymyxin B, polymyxin E, fluoxetine, diphenhydramine, clenbuterol and ketoprofen exhibited moderate toxicity towards D. magna. Additionally, three compounds (cefotaxime, polymyxin B, polymyxin E) also showed a moderate toxic effect on A. fischeri. The comparison of such results with model estimations showed inaccuracy in the predicted data, highlighting the relevance of experimental ecotoxicological assays. The assayed mixtures contained four selected drugs of high-hazard according to their reported concentrations in wastewater and surface water (diphenhydramine, trimethoprim, ketoprofen, and fluoxetine); data revealed interactions only in the fluoxetine-containing mixtures for D. magna, while all mixtures showed interactions (mostly synergistic) for Microtox. Chronic effects on the reproduction of D. magna were observed after exposure to fluoxetine and diphenhydramine, although higher sensitivity was determined for the latter, while the mixture of these compounds (which showed acute synergy in both models) also affected the reproduction patterns. Nonetheless, all the effects described at the acute or chronic level (for individual compounds or mixtures) were determined at concentrations higher than commonly reported at environmental levels. This work provides valuable ecotoxicological information for the risk assessment of pharmaceuticals and their mixtures in the environment.

Outbreak of Intermediate Species Leptospira venezuelensis Spread by Rodents to Cows and Humans in L. interrogans-Endemic Region, Venezuela.

Leptospirosis is a common but underdiagnosed zoonosis. We conducted a 1-year prospective study in La Guaira State, Venezuela, analyzing 71 hospitalized patients who had possible leptospirosis and sampling local rodents and dairy cows. Leptospira rrs gene PCR test results were positive in blood or urine samples from 37/71 patients. Leptospira spp. were isolated from cultured blood or urine samples of 36/71 patients; 29 had L. interrogans, 3 L. noguchii, and 4 L. venezuelensis. Conjunctival suffusion was the most distinguishing clinical sign, many patients had liver involvement, and 8/30 patients with L. interrogans infections died. The Leptospira spp. found in humans were also isolated from local rodents; L. interrogans and L. venezuelensis were isolated from cows on a nearby, rodent-infested farm. Phylogenetic clustering of L. venezuelensis isolates suggested a recently expanded outbreak strain spread by rodents. Increased awareness of leptospirosis prevalence and rapid diagnostic tests are needed to improve patient outcomes.

Occurrence of pharmaceuticals in Latin America: case study on hazard assessment and prioritization in Costa Rica.

Pharmaceuticals are considered as contaminants of emerging concern, and their occurrence in diverse environmental matrices has been described during the last 25 years. Nonetheless, pharmaceutical occurrence has not been evenly described worldwide, and reports from some geographical areas such as most parts of Latin America are scarce. This work aims to address the situation of water pollution due to pharmaceuticals in Latin America by means of two main goals: i. First, reviewing the monitoring studies performed in Latin America on this topic (period 2009-2024), which were conducted in Brazil, Mexico, Colombia, Ecuador, Peru and Argentina, to highlight the most frequently detected compounds from each therapeutic group in the region. ii. Second, analyzing the case of Costa Rica through the hazard assessment and prioritization of pharmaceuticals based on the monitoring performed in this country (years 2011; 2018-2019). The monitoring in Costa Rica comprised a total of 163 sampling points: wastewater treatment plants (WWTPs) (14 urban WWTPs plus two landfill WWTPs; total samples n = 44 influents and n = 34 effluents), nine hospital effluents (n = 32), wastewater from livestock farms (six swine farms and seven dairy farms; n = 23 influents and n = 37 effluents), 64 continental surface water sampling points (n = 137), and 61 coastal seawater sampling points (n = 61). Risk assessment of detected concentrations by the hazard quotient (HQ) approach (period 2018-2019) revealed a total of 25 medium or high-hazard compounds (out of 37 detected compounds). The prioritization approach (which included the Frequency of Appearance (FoA), the Frequency of PNEC exceedance (FoE), and the Extent of predicted no-effect concentration (PNEC) exceedance (EoE)), showed a critical list of nine pharmaceuticals: caffeine, diphenhydramine, acetaminophen, lovastatin, gemfibrozil, ciprofloxacin, ibuprofen, doxycycline and norfloxacin. These compounds should be taken into account as a first concern during the implementation of environmental policies related to pharmaceutical products in the region.

Improved productivity and dye removal performance of Trametes versicolor pellets using rice husk as a co-substrate.

Pellet production represents a critical step for several processes requiring fungal biomass, nevertheless, its optimization is seldom reported. The use of finely ground rice husk as a microcarrier and co-substrate permitted a marked increase (≈ 2.7×) in the productivity of fungal pellet production using Trametes versicolor compared to traditional production methods. The pellets show similar structure and smaller size compared to typical sole-mycelium pellets, as well as comparable laccase activity. The efficiency of the pellets for biodegradation was confirmed by the removal of the crystal violet dye, achieving significantly faster decolorization rates compared to the traditionally produced pellets. The use of these pellets during the continuous treatment of the dye in a stirred tank bioreactor resulted in 97% decolorization operating at a hydraulic residence time of 4.5 d.

Ecotoxicological effects of ketoprofen and fluoxetine and their mixture in an aquatic microcosm.

The effects of fluoxetine (antidepressant) and ketoprofen (analgesic) on aquatic ecosystems are largely unknown, particularly as a mixture. This work aimed at determining the effect of sublethal concentrations of both compounds individually (0.050 mg/L) and their mixture (0.025 mg/L each) on aquatic communities at a microcosm scale for a period of 14 d. Several physicochemical parameters were monitored to estimate functional alterations in the ecosystem, while model organisms (Daphnia magna, Lemna sp., Raphidocelis subcapitata) and the sequencing of 16S/18S rRNA genes permitted to determine effects on specific populations and changes in community composition, respectively. Disturbances were more clearly observed after 14 d, and overall, the microcosms containing fluoxetine (alone or in combination with ketoprofen) produced larger alterations on most physicochemical and biological variables, compared to the microcosm containing only ketoprofen, which suffered less severe changes. Differences in nitrogen species suggest alterations in the N-cycle due to the presence of fluoxetine; similarly, all pharmaceutical-containing systems decreased the brood rate of D. magna, while individual compounds inhibited the growth of Lemna sp. No clear trends were observed regarding R. subcapitata, as indirectly determined by chlorophyll quantification. The structure of micro-eukaryotic communities was altered in the fluoxetine-containing systems, whereas the structure of bacterial communities was affected to a greater extent by the mixture. The disruptions to the equilibrium of the microcosm demonstrate the ecological risk these compounds pose to aquatic ecosystems.

Genome-Wide Classification of Myb Domain-Containing Protein Families in Entamoeba invadens.

Entamoeba histolytica, the causative agent of amebiasis, is the third leading cause of death among parasitic diseases globally. Its life cycle includes encystation, which has been mostly studied in Entamoeba invadens, responsible for reptilian amebiasis. However, the molecular mechanisms underlying this process are not fully understood. Therefore, we focused on the identification and characterization of Myb proteins, which regulate the expression of encystation-related genes in various protozoan parasites. Through bioinformatic analysis, we identified 48 genes in E. invadens encoding MYB-domain-containing proteins. These were classified into single-repeat 1R (20), 2R-MYB proteins (27), and one 4R-MYB protein. The in-silico analysis suggests that these proteins are multifunctional, participating in transcriptional regulation, chromatin remodeling, telomere maintenance, and splicing. Transcriptomic data analysis revealed expression signatures of eimyb genes, suggesting a potential orchestration in the regulation of early and late encystation-excystation genes. Furthermore, we identified probable target genes associated with reproduction, the meiotic cell cycle, ubiquitin-dependent protein catabolism, and endosomal transport. In conclusion, our findings suggest that E. invadens Myb proteins regulate stage-specific proteins and a wide array of cellular processes. This study provides a foundation for further exploration of the molecular mechanisms governing encystation and unveils potential targets for therapeutic intervention in amebiasis.

Biological treatment of pesticide-containing wastewater from coffee crops: selection and optimization of a biomixture and biobed design.

The biopurification systems (BPS) or biobeds are employed for the treatment of pesticide-containing wastewater of agricultural origin. The use of these devices for pesticide removal requires the proper optimization of the composition of biomixtures (BPS active matrix) according to the target pesticides applied on a specific crop and the available materials used in their elaboration. This work aims to design a biomixture for the simultaneous treatment of several pesticides applied in coffee crops, according to local practices in Costa Rica. Three biomixtures containing either coffee husk, coconut fiber or rice husk (as the lignocellulosic substrate) were applied for the removal of 12 pesticides. The profiles of pesticide elimination and the mineralization of radiolabeled chlorpyrifos (14C-chlorpyrifos) revealed that the best performance was achieved with the coconut fiber biomixture, even though similar detoxification patterns were determined in every biomixture (according to immobilization in Daphnia magna and germination tests in Lactuca sativa). The optimization of this biomixture's composition by means of a central composite design permitted the definition of two optimal compositions (compost:soil:coconut fiber, % v/v) that maximized pesticide removal: i. 29:7.3:63.7 and ii. 11:7.3:81.7. The validation of these optimized compositions also included the use of an alternative soil from another coffee farm and resulted in overall DT50 values of 7.8-9.0 d for the pesticide mixture. Considering the removal kinetics in the optimized biomixture, a 1 m3 BPS prototype was dimensioned to be eventually used in local coffee farms. This work provides relevant information for the design and implementation of BPS at on-farm conditions for the treatment of pesticide-containing wastewater of a major crop.

Pharmaceuticals, hazard and ecotoxicity in surface and wastewater in a tropical dairy production area in Latin America.

Pharmaceuticals comprise a complex group of emerging pollutants. Despite the significant number of pharmaceuticals used in veterinary medicine, the input of these compounds into the environment due to livestock activities has been scarcely described. This work assays for the first time in Central America the occurrence of pharmaceuticals in farm wastewater in an area devoted to dairy production, and in the surrounding surface waters. Among 69 monitored pharmaceuticals, a total of eight compounds were detected in wastewater samples collected from seven dairy farms after three sampling campaigns. Six pharmaceuticals were considered either of high (albendazole, lovastatin and caffeine) or intermediate estimated hazard (ciprofloxacin, acetaminophen and ketoprofen) based on the HQ approach, while 26% of the samples were considered of high estimated hazard according to the cumulative ∑HQ approach. Similarly, when ecotoxicological tests were applied, all the samples showed some level of toxicity towards Daphnia magna, and most samples towards Vibrio fischeri and Lactuca sativa. Fourteen pharmaceuticals were detected in surface water samples collected in the surroundings of the dairy production farms, including rural and urban areas. Seven out of these compounds showed high estimated risk (risperidone, diphenhydramine, trimethoprim, fluoxetine, ofloxacin, caffeine and ibuprofen), while three (gemfibrozil, ciprofloxacin and cephalexin) exhibited intermediate estimated risk. In a similar worrisome way, 27% of these samples were estimated to pose high environmental risk according to the pharmaceutical content. Despite being nontoxic for D. magna or V. fischeri, frequent inhibition (>20%) of GI in L. sativa was determined in 34% of surface water samples; such findings raise concern on the apparent inceptive environmental pollution and risk within the area. According to the pharmaceutical content patterns in both kinds of studied matrices, no clear evidence of significant contamination in surface water due to livestock activities could be retrieved, suggesting a main role of urban influence.

Dealing with missing data under stratified sampling designs where strata are study domains.

A quick count seeks to estimate the voting trends of an election and communicate them to the population on the evening of the same day of the election. In quick counts, the sampling is based on a stratified design of polling stations. Voting information is gathered gradually, often with no guarantee of obtaining the complete sample or even information in all the strata. However, accurate interval estimates with partial information must be obtained. Furthermore, this becomes more challenging if the strata are additionally study domains. To produce partial estimates, two strategies are proposed: (1) a Bayesian model using a dynamic post-stratification strategy and a single imputation process defined after a thorough analysis of historic voting information; additionally, a credibility level correction is included to solve the underestimation of the variance and (2) a frequentist alternative that combines standard multiple imputation ideas with classic sampling techniques to obtain estimates under a missing information framework. Both solutions are illustrated and compared using information from the 2021 quick count. The aim was to estimate the composition of the Chamber of Deputies in Mexico.

SPH simulations and experimental investigation of water flow through a Venturi meter of rectangular cross-section.

The flow of water through a horizontal small-scale Venturi tube of rectangular cross-section is simulated using a modified version of the open-source code DualSPHysics, which is based on Smoothed Particle Hydrodynamics (SPH) methods. Water is simulated using the Murnaghan-Tait equation of state so that weak compressibility is allowed. The hydrodynamics is coupled to a Large-Eddy Simulation (LES) turbulence model. The convergence properties of SPH are improved by adopting a C[Formula: see text] Wendland function as the interpolation kernel, increased number of neighboring particles and non-reflective open boundary conditions at the outlet of the Venturi tube. The flow structure and differential pressure as well as the mainstream velocity profiles at different stations are compared with calibrated experimental data. A resolution independence test shows that good convergence to the experimental measurements is achieved using four million particles. At this resolution the simulations predict the experimental centerline velocity profile along the Venturi meter for a volumetric flow rate of ten liters per minutes (lpm) with a root-mean-square error of 4.3%. This error grows to 7.1% when the volumetric flow rate increases to 25 lpm. The predicted differential pressure matches the experimental data with errors varying from 1.4% (for 10 lpm) to 6.8% (for 25 lpm). Cross-sectional velocity profiles within the throat and divergent sections differ from the experimental measurements in less than 5.5%. In general, it is shown that the SPH model can provide an efficient and accurate method for recalibrating flow meters at moderately high Reynolds numbers instead of using costly experimental tests.

On-farm Occurrence of Pharmaceuticals and Their Environmental Hazard: Case Study of a Tropical Dairy farm.

Animal husbandry wastewaters represent an important source of pharmaceuticals into the environment. This work aimed to evaluate the occurrence of pharmaceuticals and their hazard in wastewater from a model dairy farm from Costa Rica. Among the seven pharmaceuticals detected (acetaminophen, caffeine, carbamazepine, ibuprofen, ketoprofen, risperidone, sulfamethazine), caffeine, ibuprofen and acetaminophen showed the highest concentrations, while caffeine, carbamazepine and risperidone were the most frequently detected compounds. High (HQ ≥ 1) or medium (0.1 ≤ HQ < 1) hazard were estimated for three (caffeine, ibuprofen, risperidone) and two (acetaminophen, ketoprofen) pharmaceuticals, respectively; similarly, high overall hazard (∑HQ) and significant ecotoxicity were determined in samples from all sampling points. According to our results, the release of these aqueous matrices is a matter of environmental concern, as the treated wastewater is used for farm irrigation or directly released into nearby water streams. This work contributes to the knowledge on the scarcely described occurrence and risk of pharmaceuticals in Latin American regions.

Occurrence and risk assessment of pharmaceuticals in hospital wastewater in Costa Rica.

This work aims to determine the occurrence, hazard and prioritization of pharmaceuticals from hospital wastewater in Costa Rica through the monitoring of 70 compounds and assessing their environmental risk through a hazard quotient approach (HQ). Moreover, the quantification of selected antibiotic resistance genes (ARGs) was conducted for the first time in this matrix in this geographical location. Thirty-four pharmaceuticals were detected, being caffeine, 1,7-dimethylxanthine, acetaminophen, ibuprofen, naproxen, ciprofloxacin and ketoprofen the most frequent (>50% of the samples). Eighteen pharmaceuticals exhibited high hazard (HQ ≥ 1), while five more showed medium hazard (1 > HQ ≥ 0.1). Prioritization, which also included frequency parameters, revealed caffeine, lovastatin, diphenhydramine, acetaminophen, ibuprofen, ciprofloxacin, and sildenafil as the compounds of major concern. Similarly, cumulative hazard per sample (ΣHQ) estimated high hazard towards aquatic organisms in every sample. All selected ARGs, except mcr-1 (polymyxin resistance), were detected. Among genes conferring resistance to beta-lactams, blaCTX-M and blaKPC were the most abundant, related to resistance to cephalosporins and carbapenems. Ecotoxicological evaluation showed mostly low toxicity towards Daphnia magna and Vibrio fischeri, contrary to the marked effect observed towards Lactuca sativa. These findings provide relevant and novel information on the risk posed by hospital wastewater and their pharmaceutical content in the Latin American environmental context.

Ecotoxicity of pesticide formulations and their mixtures: the case of potato crops in Costa Rica.

Despite their environmental implications, ecotoxicological information regarding pesticide mixtures is relatively scarce. This study aimed to determine the ecotoxicity of individual pesticide formulations and their mixtures (insecticides and fungicides), which are applied during the production cycle of potato, according to agricultural practices from a Latin American region in Costa Rica. Two benchmark organisms were employed: Daphnia magna and Lactuca sativa. First, the evaluation of individual formulations (chlorothalonil, propineb, deltamethrin+imidacloprid, ziram, thiocyclam and chlorpyrifos) revealed differences between available EC50 for active ingredients (a.i.) and their respective formulations toward D. magna; on the contrary, no information could be retrieved from scientific literature for comparison in the case of L. sativa. In general, acute toxicity was higher toward D. magna than L. sativa. Moreover, interactions could not be determined on L. sativa, as the chlorothalonil formulation was not toxic at high levels and the concentration-response to propineb could not be fitted to obtain an IC50 value. The commercial formulation composed of deltamethrin+imidacloprid followed the concentration addition model (when compared with parameters retrieved from individual a.i.) and the other three mixtures evaluated (I: chlorothalonil-propineb-deltamethrin+imidacloprid; II: chlorothalonil-propineb-ziram-thiocyclam; III: chlorothalonil-propineb-chlorpyrifos) produced an antagonistic effect on D. magna, thus suggesting less acute toxicity than their individual components. Subsequent chronic studies showed that one of the most toxic mixtures (II) negatively affected D. magna reproduction at sublethal concentrations indicating that this mixture poses a risk to this species if these pesticides co-exist in freshwater systems. These findings provide useful data to better estimate the impact of real agricultural practices related to the use of agrochemicals.

Chemical Optimization of CBL0137 for Human African Trypanosomiasis Lead Drug Discovery.

The carbazole CBL0137 (1) is a lead for drug development against human African trypanosomiasis (HAT), a disease caused by Trypanosoma brucei. To advance 1 as a candidate drug, we synthesized new analogs that were evaluated for the physicochemical properties, antitrypanosome potency, selectivity against human cells, metabolism in microsomes or hepatocytes, and efflux ratios. Structure-activity/property analyses of analogs revealed eight new compounds with higher or equivalent selectivity indices (5j, 5t, 5v, 5w, 5y, 8d, 13i, and 22e). Based on the overall compound profiles, compounds 5v and 5w were selected for assessment in a mouse model of HAT; while 5v demonstrated a lead-like profile for HAT drug development, 5w showed a lack of efficacy. Lessons from these studies will inform further optimization of carbazoles for HAT and other indications.

Marjolin's Ulcer After Saphenectomy: A Case Report.

Chronically injured areas have the possibility of transforming into malignant tissue, with squamous cell carcinoma being the most common type. This rare entity is known as Marjolin's ulcer. Most of these ulcers derive from chronic burn wounds. This case report exhibits a rare Marjolin's ulcer that developed on a 50-year-old male with a previous saphenectomy on his left leg. The patient was brought to the operating room (OR), for excision of the ulcer with a rotation flap to correct the defect. There is still no definite treatment protocol for Marjolin's ulcer. In the present article, the most common treatments are discussed. The main takeaway of this case is the prevention of Marjolin's ulcer by timely treating ulcerative lesions.

Physiologic Targets and Modes of Action for CBL0137, a Lead for Human African Trypanosomiasis Drug Development.

CBL0137 is a lead drug for human African trypanosomiasis, caused by Trypanosoma brucei Herein, we use a four-step strategy to 1) identify physiologic targets and 2) determine modes of molecular action of CBL0137 in the trypanosome. First, we identified fourteen CBL0137-binding proteins using affinity chromatography. Second, we developed hypotheses of molecular modes of action, using predicted functions of CBL0137-binding proteins as guides. Third, we documented effects of CBL0137 on molecular pathways in the trypanosome. Fourth, we identified physiologic targets of the drug by knocking down genes encoding CBL0137-binding proteins and comparing their molecular effects to those obtained when trypanosomes were treated with CBL0137. CBL0137-binding proteins included glycolysis enzymes (aldolase, glyceraldehyde-3-phosphate dehydrogenase, phosphofructokinase, phosphoglycerate kinase) and DNA-binding proteins [universal minicircle sequence binding protein 2, replication protein A1 (RPA1), replication protein A2 (RPA2)]. In chemical biology studies, CBL0137 did not reduce ATP level in the trypanosome, ruling out glycolysis enzymes as crucial targets for the drug. Thus, many CBL0137-binding proteins are not physiologic targets of the drug. CBL0137 inhibited 1) nucleus mitosis, 2) nuclear DNA replication, and 3) polypeptide synthesis as the first carbazole inhibitor of eukaryote translation. RNA interference (RNAi) against RPA1 inhibited both DNA synthesis and mitosis, whereas RPA2 knockdown inhibited mitosis, consistent with both proteins being physiologic targets of CBL0137. Principles used here to distinguish drug-binding proteins from physiologic targets of CBL0137 can be deployed with different drugs in other biologic systems. SIGNIFICANCE STATEMENT: To distinguish drug-binding proteins from physiologic targets in the African trypanosome, we devised and executed a multidisciplinary approach involving biochemical, genetic, cell, and chemical biology experiments. The strategy we employed can be used for drugs in other biological systems.

Single and mixture toxicity of selected pharmaceuticals to the aquatic macrophyte Lemna minor.

Plants represent uncommon targets to evaluate pharmaceuticals toxicity. In this work, Lemna minor was employed as a plant model to determine the toxicity of selected pharmaceuticals, and to assay if such toxicity could be predicted by QSAR models based on green algae. Among eight compounds, measurable toxicity was determined for ketoprofen (EC50 = 11.8 ± 1.9 mg/L), fluoxetine (EC50 = 27.0 ± 8.7 mg/L) and clindamycin 2-phosphate (EC50 = 57.7 ± 1.7 mg/L). Even though a correlation of r2 = 0.87 was observed between experimental toxicity towards algae and L. minor, QSAR estimations based on algae data poorly predicted the toxicity of pharmaceuticals on the plant. More experimental data for L. minor are necessary to determine the applicability of these predictions; nonetheless, these results remark the importance of measuring experimental ecotoxicological parameters for individual taxa. The toxicity of pharmaceutical binary mixtures (ketoprofen, fluoxetine and clindamycin) revealed in some cases deviations from the concentration addition model; nonetheless these deviations were small, thus the interactions are unlikely to be of severe biological significance. Moreover, the EC50 concentrations determined for these pharmaceuticals are significantly higher than those detected in the environment, suggesting that acute effects on L. minor would not take place at ecosystem level.

COVID-19 clinical footprint to infer about mortality.

Information on 4.1 million patients identified as COVID-19 positive in Mexico is used to understand the relationship between comorbidities, symptoms, hospitalisations and deaths due to the COVID-19 disease. Using the presence or absence of these variables a clinical footprint for each patient is created. The risk, expected mortality and the prediction of death outcomes, among other relevant quantities, are obtained and analysed by means of a multivariate Bernoulli distribution. The proposal considers all possible footprint combinations resulting in a robust model suitable for Bayesian inference. The analysis is carried out considering the information on the monthly COVID-19 cases, from March 2020 to the first days of January 2022. This allows one to appreciate the evolution of the mortality risk over time and the effect the strategies of the health authorities have had on it. Supporting information for this article, containing code and the dataset used for the analysis, is available online.

Recombinant Protein Expression and Purification of N, S1, and RBD of SARS-CoV-2 from Mammalian Cells and Their Potential Applications.

The coronavirus disease 2019 (COVID-19) pandemic has reached an unprecedented level. There is a strong demand for diagnostic and serological supplies worldwide, making it necessary for countries to establish their own technologies to produce high-quality biomolecules. The two main viral antigens used for the diagnostics for severe acute respiratory syndrome coronavirus (SARS-CoV-2) are the structural proteins spike (S) protein and nucleocapsid (N) protein. The spike protein of SARS-CoV-2 is cleaved into S1 and S2, in which the S1 subunit has the receptor-binding domain (RBD), which induces the production of neutralizing antibodies, whereas nucleocapsid is an ideal target for viral antigen-based detection. In this study, we designed plasmids, pcDNA3.1/S1 and pcDNA3.1/N, and optimized their expression of the recombinant S1 and N proteins from SARS-CoV-2 in a mammalian system. The RBD was used as a control. The antigens were successfully purified from Expi293 cells, with high yields of the S1, N, and RBD proteins. The immunogenic abilities of these proteins were demonstrated in a mouse model. Further, enzyme-linked immunosorbent assays with human serum samples showed that the SARS-CoV-2 antigens are a suitable alternative for serological assays to identify patients infected with COVID-19.

Environmental monitoring and risk assessment in a tropical Costa Rican catchment under the influence of melon and watermelon crop pesticides.

A monitoring network was established in streams within a catchment near the Costa Rican Pacific coast (2008-2011) to estimate the impact of pesticides in surface water (84 samples) and sediments (84 samples) in areas under the influence of melon and watermelon production. A total of 66 (water) and 47 (sediment) pesticides were analyzed, and an environmental risk assessment (ERA) was performed for four taxa (algae, Daphnia magna, fish and Chironomus riparius). One fungicide and seven insecticides were detected in water and/or sediment; the fungicide azoxystrobin (water) and the insecticide cypermethrin (sediments) were the most frequently detected pesticides. The insecticides endosulfan (5.76 µg/L) and cypermethrin (301 µg/kg) presented the highest concentrations in water and sediment, respectively. The ERA revealed acute risk in half of the sampling points of the melon-influenced area and in every sampling point from the watermelon-influenced area. Safety levels were exceeded within and around the crop fields, suggesting that agrochemical contamination was distributed along the catchment, with potential influence of nearby crops. Acute risk was caused by the insecticides chlorpyrifos, cypermethrin and endosulfan to D. magna, fish and C. riparius; the latter was the organism with the overall highest/continuous risk. High chronic risk was determined in all but one sampling point, and revealed a higher number of pesticides of concern. Cypermethrin was the only pesticide to pose chronic risk for all benchmark organisms. The results provide new information on the risk that tropical crops pose to aquatic ecosystems, and highlight the importance of including the analysis of sediment concentrations and chronic exposure in ERA.