Eight rules for improving the quality of papers on the antimicrobial susceptibility of bacteria isolated from aquatic animals.

The methods for antimicrobial susceptibility testing of bacteria, although relatively simple, are not robust. As a consequence, if the data generated in such tests are to be commensurate, all susceptibility tests must be performed using standard protocols. A review of the published literature of antimicrobial susceptibility testing of bacteria from aquatic animals revealed a frequent occurrence of significant errors, particularly with regard to testing methodology, quality control and the use of appropriate interpretive criteria in the performance and reporting of susceptibility tests. This opinion piece provides a set of rules that, if followed, would help authors to avoid these frequently detected shortcomings.

Assessing the performance of a Fasciola gigantica serum antibody ELISA to estimate prevalence in cattle in Cameroon.

BACKGROUND: Cattle rearing in Cameroon is both economically and culturally important, however parasitic diseases detrimentally impact cattle productivity. In sub-Saharan Africa bovine fasciolosis is generally attributed to F. gigantica, although understanding of Fasciola species present and local epidemiology in individual countries is patchy. Partly limited by the lack of representative surveys and understanding of diagnostic test perfromance in local cattle populations. The aims of this paper were to determine the Fasciola species infecting cattle, develop a species specific serum antibody ELISA, assess the performance of the ELISA and use it to assess the prevalence of F. gigantica exposure in two important cattle-rearing areas of Cameroon. RESULTS: A random sample of Fasciola parasites were collected and were all identified as F. gigantica (100%, CI:94.0-100%, n = 60) using RAPD-PCR analysis. A F. gigantica antibody ELISA was developed and initially a diagnostic cut-off was determined using a sample of known positive and negative cattle. The initial cut-off was used as starting point to estimate an optimal cut-off to estimate the best combination of sensitivity and specificity. This was achieved through sampling a naturally infected population with known infection status (cattle slaughtered at Bamenda abattoir, North West Region (n = 1112) and Ngaoundere abattoir, Vina Division, Adamawa Region (n = 776) in Cameroon). These cattle were tested and results analysed using a Bayesian non-gold standard method. The optimal cut-off was 23.5, which gave a sensitivity of 65.3% and a specificity of 65.2%. The prevalence of exposure to F. gigantica was higher in cattle in Ngaoundere (56.4% CI: 50.2-60.0%) than Bamenda (0.6% CI: 0.0-1.4%). CONCLUSION: Fasciola gigantica was identified as the predominant Fasciola species in Cameroon. Although the sensitivity and specificity F. gigantica antibody ELISA requires improvement, the test has shown to be a potentially useful tool in epidemiological studies. Highlighting the need for better understanding of the impact of F. gigantica infections on cattle production in Cameroon to improve cattle production in the pastoral systems of Central-West Africa. This paper also highlights that non-gold standard latent class methods are useful for assessing diagnostic test performance in naturally-infected animal populations in resource limited settings.

"In the Course of Routine Analysis": Re-Envisioning Research in State Departments of Health, 1920-1940.

During the interwar years, 1919-1939, State Department of Health (DOH) laboratories from around the United States provided scientific leadership and dedicated bench work to support the development of standardized laboratory technique. One important focus of the work was syphilis testing. In recent years, historians have explored the heated arguments and international debates surrounding the development of standardized methods. This article adds State Departments of Health to the venues in which scientists performed valuable research aimed at standardizing and improving analytic procedures. I argue that the research conducted in DOH laboratories to provide Progressive Era public health officials with accurate and efficient tools to control syphilis and other infectious diseases required significant innovation, international cooperation, and scientific creativity from scientists deeply engaged in multiple research projects. These endeavors offer an instructive venue for historians to begin to re-envision DOH laboratories as vibrant sites for scientific research, where work on standard methods was an enterprise of discovery and innovation.

Evaluation of Chronic Effects of Potassium Chloride and Nickel on Survival, Growth, and Reproduction of a Unionid Mussel (Lampsilis siliquoidea).

The ASTM International standard test method for freshwater mussels (E2455-13) recommends 4-week toxicity testing with juveniles to evaluate chronic effects on survival and growth. However, concerns remain that the method may not adequately address the sensitivity of mussels to longer term exposures (>4 weeks), particularly in relation to potential reproductive impairments. No standard method directly evaluates toxicant effects on mussel reproduction. The objectives of the present study were to (1) evaluate toxicity endpoints related to reproduction in fatmucket (Lampsilis siliquoidea) using two common reference toxicants, potassium chloride (KCl) and nickel (Ni); (2) evaluate the survival and growth of juvenile fatmucket in standard 4-week and longer term (12-week) KCl and Ni tests following a method refined from the standard method; and (3) compare the sensitivity of the reproductive endpoints with the endpoints obtained from the juvenile mussel tests. Reproductive toxicity tests were conducted by first exposing female fatmucket brooding mature larvae (glochidia) to five test concentrations of KCl and Ni for 6 weeks. Subsamples of the glochidia were then removed from the adults to determine three reproductive endpoints: (1) the viability of brooded glochidia; (2) the viability of free glochidia in a 24-h exposure to the same toxicant concentrations as their mother; and (3) the success of glochidia parasitism on host fish. Mean viability of brooded glochidia was significantly reduced in the high KCl concentration (26 mg K/L) relative to the control, with a 20% effect concentration (EC20) of 14 mg K/L, but there were no significant differences between the control and any Ni treatment (EC20 > 95 µg Ni/L). The EC20s for viability of free glochidia after the additional 24-h exposure and parasitism success were similar to the EC20s of brooded glochidia. The EC20s based on the most sensitive biomass endpoint in the 4-week juvenile tests were 15 mg K/L and 91 µg Ni/L, similar to or greater than the EC20s from the reproductive KCl and Ni tests, respectively. When exposure duration in the juvenile tests was extended from 4 to 12 weeks, the EC20s decreased by more than 50% in the KCl test but by only 8% in the Ni test. Overall, these results indicate that a standard 4-week test with juvenile mussels can prove effective for estimating effects in chronic exposures with different life stages although a longer term 12-week exposure with juvenile mussels may reveal higher sensitivity of mussels to some toxicants, such as KCl. Environ Toxicol Chem 2024;43:1097-1111. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Comparison of Cefotaxime-Resistant Escherichia coli and sul1 and intI1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water.

Awareness of the need for surveillance of antimicrobial resistance (AMR) in water environments is growing, but there is uncertainty regarding appropriate monitoring targets. Adapting culture-based fecal indicator monitoring to include antibiotics in the media provides a potentially low-tech and accessible option, while quantitative polymerase chain reaction (qPCR) targeting key genes of interest provides a broad, quantitative measure across the microbial community. The purpose of this study was to compare findings obtained from the culture of cefotaxime-resistant (cefR) Escherichia coli with two qPCR methods for quantification of antibiotic resistance genes across wastewater, recycled water, and surface waters. The culture method was a modification of US EPA Method 1603 for E. coli, in which cefotaxime is included in the medium to capture cefR strains, while qPCR methods quantified sul1 and intI1. A common standard operating procedure for each target was applied to samples collected by six water utilities across the United States and processed by two laboratories. The methods performed consistently, and all three measures reflected the same overarching trends across water types. The qPCR detection of sul1 yielded the widest dynamic range of measurement as an AMR indicator (7-log versus 3.5-log for cefR E. coli), while intI1 was the most frequently detected target (99% versus 96.5% and 50.8% for sul1 and cefR E. coli, respectively). All methods produced comparable measurements between labs (p < 0.05, Kruskal-Wallis). Further study is needed to consider how relevant each measure is to capturing hot spots for the evolution and dissemination of AMR in the environment and as indicators of AMR-associated human health risk.

Review and Development of Best Practices for Toxicity Tests with Dreissenid Mussels.

Since their introduction to North America in the 1980s, research to develop effective control tools for invasive mussels (Dreissena polymorpha and D. rostriformis bugensis) has been ongoing across various research institutions using a range of testing methods. Inconsistencies in experimental methods and reporting present challenges for comparing data, repeating experiments, and applying results. The Invasive Mussel Collaborative established the Toxicity Testing Work Group (TTWG) in 2019 to identify "best practices" and guide development of a standard framework for dreissenid mussel toxicity testing protocols. We reviewed the literature related to laboratory-based dreissenid mussel toxicity tests and determined the degree to which standard guidelines have been used and their applicability to dreissenid mussel testing. We extracted detailed methodology from 99 studies from the peer-reviewed and gray literature and conducted a separate analysis for studies using presettlement and postsettlement mussels. We identified specific components of methods and approaches that could be refined or standardized for dreissenid mussels. These components included species identification, collection methods, size/age class distinction, maintenance practices, testing criteria, sample size, response measures, reporting parameters, exposure methods, and mortality criteria. We consulted experts in the field of aquatic toxicology and dreissenid mussel biology on our proposed. The final recommendations contained in the present review are based on published standard guidelines, methods reported in the published and gray literature, and the expertise of TTWG members and an external panel. In addition, our review identifies research needs for dreissenid mussel testing including improved methods for early-life stage testing, comparative data on life stages and between dreissenid mussel species, inclusion of a reference toxicant, and additional testing of nontarget species (i.e., other aquatic organisms). Environ Toxicol Chem 2023;42:1649-1666. © 2023 His Majesty the King in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Comparison of Standardized Methods for Determining the Diffusion Coefficient of Chloride in Concrete with Thermodynamic Model of Migration.

This research paper is the result of observations made during tests according to various standards carried out on behalf of industry. The article presents diffusion coefficient values calculated according to the thermodynamic migration model for twenty different concrete mixes and some selected mixes of the codified approaches known as ASTM 1202, NT BUILD 443, NT BUILD 492, ASTM 1556. The method used here, according to the thermodynamic model of migration, allows determination of the value of the diffusion coefficient after short studies of the migration of chloride ions into concrete and was described in earlier works by one of the authors. Unfortunately, when using standard methods, the values of diffusion coefficients differ significantly from each other. In each concrete, diffusion tests were carried out in the conditions of long-term natural diffusion to verify the values determined by standard methods and according to the thermodynamic model of migration. The analysis conducted for this research paper reveals that the chloride permeability test method according to the standard ASTM C1202-97 has an almost 2.8-fold greater dispersion of the obtained results compared to the thermodynamic model of migration. It was observed that the standard NT BUILD 492 has a 3.8-fold dispersion of results compared to the method with the thermodynamic model of migration. The most time-consuming method is the standard method NT BUILD 443. The largest 3.5-fold dispersion of values concerning the reference value are observed in that method. Moreover, a method based on a thermodynamic migration model seems to be the best option of all analyzed methods. It is a quite quick, but laborious, method that should be tested for a larger number of concrete mixes. A great advantage of this method is that it is promising for a wide range of concrete mixtures, both plain concrete and concrete with various additives and admixtures, as well as high-performance concrete.

Determining MOSH and MOAH with High Sensitivity in Vegetable Oil─A New, Reliable, and Comparable Approach Using Online LC-GC-FID─Evaluation of Method Precision Data.

A method for the analysis of saturated mineral oil hydrocarbons (MOSH) and aromatic mineral oil hydrocarbons (MOAH) has been developed to improve interlaboratory precisions especially for amounts below 10 mg/kg. This approach was adopted as the standard method DGF C-VI 22 (20) of the German Society of Fat Sciences. Therefore, this method was evaluated on different edible oils containing a variety of interfering biogenic substances. The precision data were determined in an interlaboratory trial with an international group of 14 laboratories from Germany, Austria, and Italy. Good reproducible relative standard deviations for total MOSH (12.5-20.7%) and total MOAH (12.4-39.5%) as well as HorRat values ranging between 1.1 and 2.3 for total MOSH and between 0.9 and 2.6 for total MOAH have been achieved. As some matrices showed residual interferences in the MOAH fraction, these substances were further analyzed by online high-performance liquid chromatography-comprehensive two dimensional gas chromatography with time of flight mass detection.

Environmental Profile of a Novel High-Amylose Bread Wheat Fresh Pasta with Low Glycemic Index.

To improve glycemic health, a high-amylose bread wheat flour fresh pasta characterized by a low in vitro glycemic index (GI) and improved post-prandial glucose metabolism was previously developed. In this study, well-known life cycle analysis software was used in accordance with the PAS 2050 and mid- and end-point ReCiPe 2016 standard methods to assess, respectively, its carbon footprint and overall environmental profile, as weighted by a hierarchical perspective. Even if both eco-indicators allowed the identification of the same hotspots (i.e., high-amylose bread wheat cultivation and consumer use of fresh pasta), the potential consumer of low-GI foods should be conscious that the novel low-GI fresh pasta had a greater environmental impact than the conventional counterpart made of common wheat flour, their corresponding carbon footprint or overall weighted damage score being 3.88 and 2.51 kg CO2e/kg or 184 and 93 mPt/kg, respectively. This was mainly due to the smaller high-amylose bread wheat yield per hectare. Provided that its crop yield was near to that typical for common wheat in Central Italy, the difference between both eco-indicators would be not greater than 9%. This confirmed the paramount impact of the agricultural phase. Finally, use of smart kitchen appliances would help to relieve further the environmental impact of both fresh pasta products.

Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens.

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis.Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination.Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures.Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography.Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis.Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.

A Method for the Accurate Quantification of Gas Streams by Online Mass Spectrometry.

The accuracy of the online quantification of gaseous effluents from catalytic reactors by mass spectrometry (MS) is rarely addressed by researchers despite the extensive use of the technique. MS results are strongly sensitive to both the operation conditions of the reactor and to the state of the instrument. Therefore, most studies use them as qualitative descriptors of the performance of catalytic reaction systems. The purpose of this work was to develop an accurate method for the quantification of gaseous effluents from catalytic reactors. For this purpose, the mathematical expressions from the so-called external and internal standard calibration methods for MS were coupled to the typical metrics used for studying catalytic reactions, namely, conversion, selectivity, and carbon mass balances. The catalytic combustion of methane was selected as a model reaction to test the developed approach. The accuracy of the developed method was validated by comparison with results obtained in a separate reaction system coupled online to a gas chromatograph. The closure of the carbon mass balance was used as control metrics allowing for the assessment of the physical meaning of the method. In general, the internal standard method of calibration was found to be best for the accurate quantification of gaseous streams by online mass spectrometry. In general, the results of this investigation may be of use to researchers in the field of catalysis as well as to research workers using mass spectrometry for various purposes.

Sensitivities of a standard test method for the determination of the pHe of bioethanol and suggestions for improvement.

An assessment of the sensitivities of the critical parameters in the ASTM D6423 documentary standard method for the measurement of pHe in (bio)ethanol has been undertaken. Repeatability of measurements made using the same glass electrode and reproducibility between different glass electrodes have been identified as the main contributors to the uncertainty of the values produced. Strategies to reduce the uncertainty of the measurement have been identified and tested. Both increasing the time after which the pHe measurement is made following immersion in the sample, and rinsing the glass electrode with ethanol prior to immersion in the sample, have been shown to be effective in reducing the uncertainty of the numerical value produced. However, it is acknowledged that the values produced using these modified approaches may not be directly compared with those obtained using the documentary ASTM method since pHe is defined operationally by the process used to measure it.

Detection of Salmonella in Food Matrices, from Conventional Methods to Recent Aptamer-Sensing Technologies.

Rapid detection of the foodborne pathogen Salmonella in food processing is of crucial importance to prevent food outbreaks and to ensure consumer safety. Detection and quantification of Salmonella species in food samples is routinely performed using conventional culture-based techniques, which are labor intensive, involve well-trained personnel, and are unsuitable for on-site and high-throughput analysis. To overcome these drawbacks, many research teams have developed alternative methods like biosensors, and more particularly aptasensors, were a nucleic acid is used as biorecognition element. The increasing interest in these devices is related to their high specificity, convenience, and relative rapid response. This review aims to present the advances made in these last years in the development of biosensors for the detection and the quantification of Salmonella, highlighting applications on meat from the chicken food chain.

Approaches to biofilm-associated infections: the need for standardized and relevant biofilm methods for clinical applications.

INTRODUCTION: The concept of biofilms in human health and disease is now widely accepted as cause of chronic infection. Typically, biofilms show remarkable tolerance to many forms of treatments and the host immune response. This has led to vast increase in research to identify new (and sometimes old) anti-biofilm strategies that demonstrate effectiveness against these tolerant phenotypes. Areas covered: Unfortunately, a standardized methodological approach of biofilm models has not been adopted leading to a large disparity between testing conditions. This has made it almost impossible to compare data across multiple laboratories, leaving large gaps in the evidence. Furthermore, many biofilm models testing anti-biofilm strategies aimed at the medical arena have not considered the matter of relevance to an intended application. This may explain why some in vitro models based on methodological designs that do not consider relevance to an intended application fail when applied in vivo at the clinical level. Expert commentary: This review will explore the issues that need to be considered in developing performance standards for anti-biofilm therapeutics and provide a rationale for the need to standardize models/methods that are clinically relevant. We also provide some rational as to why no standards currently exist.

Thinking critically about whole-grain definitions: summary report of an interdisciplinary roundtable discussion at the 2015 Whole Grains Summit.

Definitions for whole grain (WG) have been published by governments, the food industry, and grain organizations and generally fall into 2 categories: WG and WG food. WG definitions focus on the principal components of the WGs and their proportions, whereas WG-food definitions describe the quantity of WGs present in food. In the United States, widespread agreement exists on the main parts of a definition for a WG, with a definition for a WG food still in its early stages; a standard definition that has been universally accepted does not exist. Furthermore, nutrition policy advises consumers to eat WGs for at least one-half of their total grain intake (2010 and 2015 Dietary Guidelines for Americans), but confusion exists over which foods are considered WGs and how much is needed to achieve health benefits. In December 2014, a workshop sponsored by the subcommittee on collaborative process of the US Government's Interagency Committee on Human Nutrition Research convened in Washington, DC, and recognized WG definitions as a key nutrition and public health-related issue that could benefit from further collaboration. As a follow-up to that meeting, an interdisciplinary roundtable meeting was organized at the Whole Grains Summit on 25 June 2015 in Portland, Oregon, to help resolve the issue. This article summarizes the main opportunities and challenges that were identified during the meeting for defining WGs and WG foods internationally. Definitions of WGs and WG foods that are uniformly adopted by research, food industry, consumer, and public health communities are needed to enable comparison of research results across populations.

Bridging the Gap of Standardized Animals Models for Blast Neurotrauma: Methodology for Appropriate Experimental Testing.

Recent military combat has heightened awareness to the complexity of blast-related traumatic brain injuries (bTBI). Experiments using animal, cadaver, or biofidelic physical models remain the primary measures to investigate injury biomechanics as well as validate computational simulations, medical diagnostics and therapies, or protection technologies. However, blast injury research has seen a range of irregular and inconsistent experimental methods for simulating blast insults generating results which may be misleading, cannot be cross-correlated between laboratories, or referenced to any standard for exposure. Both the US Army Medical Research and Materiel Command and the National Institutes of Health have noted that there is a lack of standardized preclinical models of TBI. It is recommended that the blast injury research community converge on a consistent set of experimental procedures and reporting of blast test conditions. This chapter describes the blast conditions which can be recreated within a laboratory setting and methodology for testing in vivo models within the appropriate environment.

Objective evaluation of reconstruction methods for quantitative SPECT imaging in the absence of ground truth.

Quantitative single-photon emission computed tomography (SPECT) imaging is emerging as an important tool in clinical studies and biomedical research. There is thus a need for optimization and evaluation of systems and algorithms that are being developed for quantitative SPECT imaging. An appropriate objective method to evaluate these systems is by comparing their performance in the end task that is required in quantitative SPECT imaging, such as estimating the mean activity concentration in a volume of interest (VOI) in a patient image. This objective evaluation can be performed if the true value of the estimated parameter is known, i.e. we have a gold standard. However, very rarely is this gold standard known in human studies. Thus, no-gold-standard techniques to optimize and evaluate systems and algorithms in the absence of gold standard are required. In this work, we developed a no-gold-standard technique to objectively evaluate reconstruction methods used in quantitative SPECT when the parameter to be estimated is the mean activity concentration in a VOI. We studied the performance of the technique with realistic simulated image data generated from an object database consisting of five phantom anatomies with all possible combinations of five sets of organ uptakes, where each anatomy consisted of eight different organ VOIs. Results indicate that the method provided accurate ranking of the reconstruction methods. We also demonstrated the application of consistency checks to test the no-gold-standard output.

Efficiency of ASTM Method D4994-89 for Recovery of Enteric Viruses from Biosolids.

The ASTM Method D4994-89 has been used in the United States for almost two decades to assess the virological quality of biosolids. However, the efficiency of this method for recovery of different enteric viruses has never been determined. The method was found to recover several different enteroviruses and adenovirus 2 with an efficiency ranging from 18.1 ± 5.5 to 24.6 ± 7.8%.

Implications of water hardness in ecotoxicological assessments for water quality regulatory purposes: a case study with the aquatic snail Biomphalaria glabrata (Say, 1818) / Implicações da dureza da água nas avaliações ecotoxicológicas com propósitos reguladores de qualidade da água: um estudo de caso com o caramujo aquático Biomphalaria glabrata

Water hardness is a property depending on the presence of alkaline earth metals, mainly calcium and magnesium. Among the strategies for water quality monitoring, ecotoxicological assays are performed to minimize impacts and classify water bodies. For these laboratory evaluations parameters are previously defined in the guidelines, including water hardness for both cultivation and testing medium. The present work was performed to evaluate the effects of different levels of water hardness on the survival and reproduction of the freshwater snail Biomphalaria glabrata and discuss the influence of natural water hardness on the results of ecotoxicological tests with these environmental samples. Comparing the groups it was possible to observe that those maintained in waters with least hardness had lower reproductive success, while the groups maintained in highest hardness showed better reproduction. These data show that waters with low hardness make the reproduction of the snail B. glabrata unfeasible, and this reveal a problem for ecotoxicity assays using natural water samples.

A dureza da água é uma propriedade dependente da presença de metais alcalino terrosos, principalmente cálcio e magnésio. Entre as estratégias para monitorar a qualidade da água ensaios ecotoxicológicos são realizados para minimizar impactos e classificar os corpos hídricos. Para essas avaliações em laboratório, parâmetros são previamente definidos nos protocolos, incluindo a dureza da água para cultivo e para a água de diluição. O presente trabalho foi realizado para avaliar os efeitos de diferentes níveis de dureza da água sobre a sobrevivência e a reprodução do caramujo de água doce Biomphalaria glabrata e discutir a influência da dureza de águas naturais nos resultados dos testes ecotoxicológicos com estas amostras ambientais. Comparando os grupos foi possível observar que aqueles mantidos em águas com menor dureza tiveram pior sucesso reprodutivo, enquanto os mantidos nas águas com maiores durezas tiveram melhor reprodução. Esses dados mostram que águas com baixas durezas tornam a reprodução do caramujo B. glabrata inviável, e esse fato revela-se como um problema para os ensaios ecotoxicológicos utilizando amostras de água naturais.