Transcriptomic signaling in zebrafish (Danio rerio) embryos exposed to environmental concentrations of glyphosate.

Glyphosate [N-(phosphonomethyl)glycine] is one of the most popular herbicides worldwide. Globally, the use of glyphosate is increasing, and its residues have been found in drinking water and food products. The data regarding the possible toxic effects of this herbicide are controversial. Therefore, the aim of this study was to evaluate the effects of glyphosate at environmental concentrations in zebrafish (Danio rerio) embryos. Embryos were exposed to 0, 1, 100, and 1,000 µg/L glyphosate for 96 h, and mortality, heart rate, and hatching rate were evaluated. After the experiment, RNA was extracted from the embryos for transcriptional analysis. No mortality was recorded, and exposure to 100 µg/L and 1,000 µg/L of glyphosate resulted in lower heart rates at 48 h. In addition, RNA-seq analysis revealed that glyphosate exposure induced subtle changes in gene transcription profiles. We found 30 differentially expressed genes; however, the highest glyphosate concentration (1,000 µg/L) induced the greatest number of differentially expressed genes involved in oocyte maturation, metabolic processes, histone deacetylation, and nervous system development.

First Evidence of Glyphosate in American Horseshoe Crab from the Yucatan Peninsula in Mexico.

Glyphosate is the most used herbicide in the world. Unfortunately, contamination of water bodies by this herbicide has been reported. A severe concern has been triggered given its detrimental impact on the environment and wildlife. The American horseshoe crab (Limulus polyphemus) is a benthic arthropod that inhabits the Yucatan Peninsula in Southeast Mexico. This study evaluates the glyphosate concentration in 34 recently dead specimens of L. polyphemus from four localities of the Ria Lagartos Biosphere Reserve in Yucatan, Mexico. The analysis was carried out using High-Performance Liquid Chromatography coupled with a Triple Quadrupole Mass Spectrometer. All the samples showed residues of glyphosate in the range from 0.08 to 2.38 ng g-1. These records constitute the first evidence of glyphosate bioaccumulation in this species. Although the scope might be limited, the results demonstrate a potentially prejudicial exposition of the marine biota to glyphosate-based herbicides, given its use in the region.

The Development and Evaluation of Brain and Heart Cell Lines from a Marine Fish for Use in Xenobiotic-Induced Cytotoxicity Testing.

Two cell lines derived from the brain and heart of a Pacific white snook specimen (Centropomus viridis) were developed and evaluated in terms of their responsiveness to glyphosate-induced cytotoxicity. The cells were grown in Leibovitz-15 (L-15) medium supplemented with 10% fetal bovine serum (FBS) and were passaged 36 times. Growth was tested at different concentrations of FBS (5, 10 and 20%) at 27°C. The cell lines were cryopreserved at different passages and were successfully thawed, with a survival rate greater than 80% without detectable contamination. At passage 36, the cells were used to assess the deleterious effects of glyphosate, and cell proliferation was measured by direct counting and with the MTT assay. Similar LC50 values were obtained with both methods. Although the principles behind these two assessment methods differ, our results show that both are suitable for evaluating glyphosate toxicity. In addition, heart- and brain-derived cells showed similar sensitivity, suggesting that the same mode of action might be responsible for the toxicity of glyphosate at the cellular level. The newly developed Pacific white snook brain and heart cell lines could be useful to investigate cellular and molecular mechanisms of toxicity, satisfying the need to reduce the use of animals in experiments. Glyphosate-related toxicological data obtained in the present study will allow us to continue investigating the effects of this herbicide directly on brain and heart fish cells since similar studies have only been carried out on either live organisms or on human cell lines such as neuroblastoma, which are immortalised by oncogenes or similar.

Multiple biomarker approach in the fiddler crab to assess anthropogenic pollution in tropical coastal environments.

CONTEXT: Fiddler crabs are important to the ecology of estuarine systems around the world, however, few studies have incorporated them as bioindicators. Urias estuary represents one of the most urbanized lagoons in the Gulf of California region and received discharges from different sources: shrimp farm, thermoelectric plant, fish processing plants, and untreated domestic and sewage wastes. OBJECTIVE: Assess the effects on anthropogenic contamination on female fiddler crabs reproduction, survival and genetic stability. METHODS: Exposition of wild crabs from a less impacted (reference) site to naturally contaminated sediments on under controlled laboratory conditions. Reproductive parameters, levels of DNA damage and mortality rates were measured, together with chemical analyses of sediments. RESULTS: The most contaminated sediments corresponded to the site where fish processing plants were located and the integrated biomarker response analysis revealed that the most adverse effects were produced by exposure to sediments from this site; these crabs showed higher mortality (67%) and poorer ovarian development than those crabs exposed to sediments from other sites. CONCLUSIONS: Female crabs under pollution stress are able to trade-off reproduction for survival, and surviving animals were able to restore genetic stability possibly by activating DNA repair mechanisms. Multiple biomarker approach discriminates different coastal contamination scenarios.

The genus Cletocamptus (Harpacticoida, Canthocamptidae): a reappraisal, with proposal of a new subfamily, a new genus, and a new species.

A new species of Cletocamptus closely related to C.helobius was found in sediment samples taken from a polluted estuarine system in north-western Mexico. The genus Cletocamptus was relegated to species incertae sedis in 1986, and this finding prompted us to evaluate the current taxonomic position of the genus within the Canthocamptidae. The latter has been subdivided in several, seemingly unnatural subfamilies in the past to better understand the relationships between its constituent taxa. In this study we propose a new subfamily, the Cletocamptinaesubfam. nov. for Amphibiperita, Cletocamptus, and Cletocamptoides gen. nov., defined by the synapomorphic subdistal ventral spinules on the rostrum. The genus Cletocamptoides gen. nov. is proposed for C.helobius, C.merbokensis, and C.biushelo sp. nov., and is supported by the 'cletodid' shape of the body and the reduced one-segmented endopod of the fourth swimming leg. Cletocamptus includes all the other species with long slender spinules on the posterior margin of prosomites and with the sexually modified inner spine on the second endopodal segment of the second swimming leg in the males. Amphibiperita retained the primitive female fifth leg with exopod and baseoendopod separated, and the primitive prehensile endopod of the first leg, but is defined by the loss of the antennary exopod. Other (syn)apomorphies are given, and the evolution of the mandibular palp is briefly discussed. Additionally, a diagnosis for the new subfamily, Cletocamptinae subfam. nov., an amended narrower diagnosis for Cletocamptus, the diagnosis for Cletocamptoides gen. nov., and a phylogenetic analysis supporting the proposal of these new taxa, are given.

Non-native terrestrial slugs from Sinaloa, Mexico: Deroceraslaeve (O. F. Müller, 1774) and Sarasinulaplebeia (P. Fischer, 1868) (Mollusca, Gastropoda).

This is the first record of two non-native terrestrial slug species from Sinaloa, Mexico. Deroceraslaeve and Sarasinulaplebeia were collected between 2019 and 2022 in Concordia and Mazatlan Municipalities (north-western Mexico). The external morphology and anatomic features of the dissected specimens coincide with the descriptions of each species, whose identities were also confirmed by their partial COI sequences. The ample occurrence of S.plebeia suggests that this species has an established population, while D.laeve was found as isolated individuals, likely associated with plant nurseries.

Feeding behavior of early life stages of the zebrafish Danio rerio is altered by exposure to glyphosate.

Glyphosate levels and the transfer of glyphosate across trophic levels have rarely been studied in zooplankton. The food preferences of zebrafish during the first-feeding stage (which is critical for the survival of organisms), were analyzed because of the requirement for live food. Larval survival begins to be affected when glyphosate intake exceeds 0.3666 µg/larvae/day, in the case that only the food is contaminated; if the medium is also contaminated, the effects on survival start from 0.2456 µg/larvae/day. It was shown that glyphosate was more likely to be incorporated through the medium than through the food (zooplankton), which supports the results of previous studies that have ruled out the potential for biomagnification. The bioconcentration factor (BCF) of glyphosate was determined using an ELISA tests specific to measure glyphosate in the fish D. rerio, the rotifers Brachionus calyciflorus and Lecane papuana, and the cladoceran Ceriodaphnia dubia. The experimental design consisted in exposing seven zebrafish adults per replica (four replicates) in three treatments 1, 5, and 10 mg/L of glyphosate for 96 h to obtain bioconcentration factors in the gills, liver, and muscle. These concentrations were selected as potential glyphosate concentrations right after application as double highest reported concentration. Glyphosate levels in zooplankton can represent up to 6.26% of the total weight of rotifers (BFC = 60.35) and in zebrafish adult organs were less than 8 µg/mg of tissue (BCF values < 6). Although glyphosate does not biomagnify, our results suggest that glyphosate affected the dynamics between zooplankton and zebrafish larvae, diminishing survival and feeding rates, given that zooplankton species bioconcentrate glyphosate in large quantities. The BCF values found in this contribution are higher than expected. Glyphosate exposure affected energy metabolism and feeding behavior of zebrafish larvae, which presented high mortality rates at environmentally relevant concentrations.

Epigenetic Transgenerational Modifications Induced by Xenobiotic Exposure in Zebrafish.

Zebrafish (Danio rerio) is a well-established vertebrate model in ecotoxicology research that responds to a wide range of xenobiotics such as pesticides, drugs, and endocrine-disrupting compounds. The epigenome can interact with the environment and transform internal and/or external signals into phenotypic responses through changes in gene transcription. Environmental exposures can also generate epigenetic variations in offspring even by indirect exposure. In this review, we address the advantages of using zebrafish as an experimental animal model to study transgenerational epigenetic processes upon exposure to xenobiotics. We focused mostly on DNA methylation, although studies on post-translational modifications of histones, and non-coding RNAs related to xenobiotic exposure in zebrafish are also discussed. A revision of the methods used to study epigenetic changes in zebrafish revealed the relevance and reproducibility for epigenetics-related research. PubMed and Google Scholar databases were consulted for original research articles published from 2013 to date, by using six keywords: zebrafish, epigenetics, exposure, parental, transgenerational, and F2. From 499 articles identified, 92 were considered, of which 14 were selected as included F2 and epigenetic mechanisms. Current knowledge regarding the effect of xenobiotics on DNA methylation, histone modifications, and changes in non-coding RNAs expressed in F2 is summarized, along with key experimental design considerations to characterize transgenerational effects.

Changes in microtubule stability in zebrafish (Danio rerio) embryos after glyphosate exposure.

Glyphosate, the most commonly used pesticide worldwide, blocks aromatic amino acid biosynthetic pathways and inhibits growth in plants. Although the specific mode of action of glyphosate in animals remains unclear, adverse effects during embryonic development have been reported, including epiboly delays, morphological alterations, and changes in central nervous system development and cardiogenesis. In this study, we suggest a possible toxicity mechanism for this herbicide related to changes in microtubule stability, which could alter the distribution and dynamics of cytoskeleton components. Using zebrafish embryos to evaluate in vivo effects of glyphosate exposure (5, 10, and 50 µg/ml), we found significant reductions in the levels of acetylated α-tubulin (50 µg/ml) and in the polymeric tubulin percentage in zebrafish embryos that had been exposed to 10 and 50 µg/ml glyphosate, without any changes in either the expression patterns of α-tubulin or the stability of actin filaments. These results indicate that high concentrations of glyphosate were associated with reduced levels of acetylated α-tubulin and altered microtubule stability, which may explain some of the neurotoxic and cardiotoxic effects that have been attributed to this herbicide.

Short exposure to glyphosate induces locomotor, craniofacial, and bone disorders in zebrafish (Danio rerio) embryos.

Glyphosate [N-(phosphonomethyl)glycine] is the active ingredient in widely used broad-spectrum herbicides. Even though the toxicity mechanism of this herbicide in vertebrates is poorly understood, evidence suggests that glyphosate is an endocrine disruptor capable of producing morphological anomalies as well as cardiotoxic and neurotoxic effects. We used the zebrafish model to assess the effects of early life glyphosate exposure on the development of cartilage and bone tissues and organismal responses. We found functional alterations, including a reduction in the cardiac rate, significant changes in the spontaneous tail movement pattern, and defects in craniofacial development. These effects were concomitant with alterations in the level of the estrogen receptor alpha osteopontin and bone sialoprotein. We also found that embryos exposed to glyphosate presented spine deformities as adults. These developmental alterations are likely induced by changes in protein levels related to bone and cartilage formation.

Genomic and molecular evolutionary dynamics of transcriptional response regulator genes in bacterial species of the Harveyi clade of Vibrio.

Transcriptional response regulators (TRR) are the most abundant signal transducers in prokaryotic systems that mediate intracellular changes in response to environmental signals. They are involved in a wide range of biological processes that allow bacteria to persist in particular habitats. There is strong evidence that the bacterial habitat and their lifestyle influence the size of their TRR genetic repertoire. Therefore, it would be expected that the evolution of bacterial genomes could be linked to natural selection processes. To test this hypothesis, we explored the evolutionary dynamics of TRR genes of the widely studied Harveyi clade of the genus Vibrio at the molecular and genomic levels. Our results suggest that the TRR genetic repertoire of the species belonging to the Harveyi clade is a product of genomic reduction and expansion. The gene loss and gains that drive their genomic reduction and expansion could be attributed to natural selection and random genetic drift. It seems that natural selection acts to maintain the ancestral state of core TRR genes (shared by all species) by purifying processes and could be driving the loss of some accessory (found in certain species) genes through the diversification of sequences. The neutrality observed in gene gain could be attributed to spontaneous events as horizontal gene transfer driven by stochastic events as occurs in random genetic drift.

Tubicolous polychaete worms (Annelida) from Bahía de Chamela Islands Sanctuary, Mexico, with the description of a new bamboo worm.

BACKGROUND: The islands and islets of Bahía de Chamela, in the Eastern Tropical Pacific, were declared as the first marine sanctuary in Mexico and has been protected since 2002. Their marine biodiversity has been documented in a series of papers in the last decade, but only three species of polychaete worms have been reported. NEW INFORMATION: Sixteen species of sedentary polychaete worms belonging to the families Maldanidae, Oweniidae, Sabellariidae, Sabellidae and Serpulidae are reported to the Bahía de Chamela Islands Sanctuary, 15 of these species constituting the first records in the area. Isocirrus tropicus (Monro, 1928) (Monro 1928) and Notaulax californica (Treadwell, 1906) (Treadwell 1906) constitute new records to Mexico; Idanthyrsus mexicanus Kirtley, 1904 (Kirtley 1994) is first recorded since its description and one species of bamboo worm (Maldanidae) is described as new to science. The new species belongs to the genus Clymenura Verril, 1900 (Verrill 1900) and its characterised by the presence of a glandular shield on chaetiger 8; a cephalic plaque oval with smooth margins and a rounded palpode; nuchal organs straight, parallel, almost full length of plaque; manubriavicular uncini present from chaetiger 1 with 3-4 teeth above the main fang without hairs or bristles; two pre-anal achaetous segments with tori; an anal funnel with alternating triangular cirri, being the longest that are located mid-ventrally.

Reestablishment of Notopygos megalops McIntosh, description of N. caribea sp. n. from the Greater Caribbean and barcoding of "amphiamerican" Notopygos species (Annelida, Amphinomidae).

The species of the genus Notopygos Grube, 1855 are characterized by an ovate body, a prominent caruncle with three lobes, dendritic branchiae, and double dorsal cirri. Twenty-two species belonging to Notopygos have been described, mostly from the Indo-Pacific region. In America, few species are frequently recorded: Notopygos crinita Grube, 1855 from St. Helena Island (Atlantic) and Notopygos ornata Grube and Ørsted in Grube 1857 from Costa Rica (Pacific). Notopygos crinita is a widely distributed species in the Western Atlantic with additional reports in the Mediterranean Sea (as a questionable alien species) and in the Pacific Ocean. However, only the genus features have been considered, consequently some records could be misidentifications. During a revision of materials from collections and the barcode project, 'Mexican Barcode of Life, MEXBOL', we found specimens of Notopygos megalops and an undescribed species from reef zones in the Caribbean; the former had been considered a junior synonym of Notopygos crinita. Herein, Notopygos megalops is reestablished and Notopygos caribeasp. n. is described. A morphological and DNA barcode approach was used to explain the records of Notopygos ornata in the Atlantic and to show the differences with the new species, since both species share features such as complex pigmentation patterns, and circular projections in the median lobe of the caruncle.

Pseudanthessius tortuosus Stock, 1967 (Copepoda: Cyclopoida) from the amphinomid polychaete Hermodice carunculata (Pallas) in the western Caribbean.

The copepod order Cyclopoida contains mainly forms that have been found as symbionts of different vertebrate and invertebrate hosts. One of these groups is the family Pseudanthessiidae, found mainly on echinoderms but very rarely on polychaetes. In this contribution, we report and confirm the occurrence of the pseudanthessiid copepod Pseudanthessius tortuosus Stock, 1967 as a symbiont of the benthic amphinomid polychaete Hermodice carunculata (Pallas) in the western Caribbean. These specimens showed subtle morphological but important size differences with respect to the specimens from the type-locality in the eastern Caribbean. Data are provided on the sex-ratio, the distribution of the copepods on the polychaete body and the finding of pairs in amplexus. The position of the copepods on the body, mainly on the dorsal branchiae, rather than on the lateral ones, allows them to avoid the friction related to the displacement mechanism of the polychaete, involving the lateral branchiae. This is the first record of the species and the symbiotic association in the western Caribbean and in Mexican waters. It is also recorded herein for the first time in the US Virgin Islands, near the type-locality. Hitherto, P. tortuosus has not been recorded since its original description forty years ago. This finding supports the hypothesis that this copepod is associated to H. carunculata throughout the entire Caribbean Basin, but its relation with other amphinomids remains unconfirmed.