José Jesús Bustillos-Guzmán

Saxitoxin (STX) is a potent aquatic neurotoxin that selectively blocks voltage-gated sodium channels (VGSCs), which play a central role in neuronal excitability. This study evaluated the behavioral effects of sublethal doses of STX in a murine model of chemically induced hyperexcitability using pentylenetetrazol (PTZ), and compared its effects with phenytoin (PHT) a conventional VGSC blocker. STX produced stage-specific and dose-dependent modulation of PTZ-induced behaviors. At 8.5 µg/kg, STX delayed the onset of early myoclonic activity. At intermediate doses, particularly 7.5 µg/kg, it reduced generalized clonic seizures (GCS) but also promoted the emergence of generalized tonic-clonic (GCTS) manifestations, suggesting a shift in excitability thresholds depending on the seizure stage. In comparison, PHT delayed the onset of myoclonic activity but was less effective than STX in reducing GCS, while more effective in completely suppressing GCTS. These differences may reflect distinct mechanisms of VGSCs modulation between both compounds. Although the highest dose of STX tested (9.5 µg/kg) was associated with marked toxicity, the ability of STX to selectively modulate excitability stages highlights its value as a neuropharmacological tool. These findings support further investigation of STX and its safer analogs in the targeted modulation of mechanisms of hyperexcitability mediated by VGSCs.

Gymnodinium catenatum, un dinoflagelado productor de toxinas paralizantes, es una de las especies de dinoflagelados más estudiadas, sin embargo, poco se conoce sobre sus interacciones con bacterias. En este estudio se determinó el efecto de un consorcio bacteriano en cepas de G. catenatum. Se emplearon 4 cepas de G. catenatum aisladas de Bahía de Santiago, Colima; Lázaro Cárdenas, Michoacán; Bahía Concepción y Bahía de La Paz, Baja California Sur. De la cepa originaria de la costa de Colima se aisló un consorcio bacteriano compuesto de 3 bacterias empleando placas de agar con medio GSe. Se incubaron a una salinidad de 34, a 24 °C y en un ciclo de 12:12 h L:O. Gymnodinum catenatum se cultivó en medio GSe líquido en las mismas condiciones que las bacterias. La interacción entre los microorganismos se evaluó inoculando el consorcio bacteriano en las cepas de G. catenatum. Se determinó la abundancia máxima, las tasas de crecimiento, la longitud de cadenas y el perfil de pigmentos y toxinas paralizantes en G. catenatum en presencia y ausencia del consorcio bacteriano. En presencia de las bacterias no se observó la misma respuesta en el crecimiento de las cepas de G. catenatum, ni en el perfil y contenido de toxinas, pero sí se observó un incremento significativo en el número de células en cadena. También se notaron cambios en la morfología de G. catenatum. Además, en presencia del consorcio bacteriano, en la mayoría de las cepas incrementó el contenido de pigmentos accesorios. Los resultados sugieren que el consorcio bacteriano pudiera ser un agente estresante para G. catenatum proporcionando nuevas perspectivas de interacción entre las bacterias y G. catenatum.

In September and November 2016, eight marine sampling sites along the coast of the southeastern Gulf of Mexico were monitored for the presence of lipophilic and hydrophilic toxins. Water temperature, salinity, hydrogen potential, dissolved oxygen saturation, inorganic nutrients and phytoplankton abundance were also determined. Two samples filtered through glass fiber filters were used for the extraction and analysis of paralytic shellfish toxins (PSTs) by lateral flow immunochromatography (IFL), HPLC with post-column oxidation and fluorescent detection (FLD) and UHPLC coupled to tandem mass spectrometry (UHPLC-MS/MS). Elevated nutrient contents were associated with the sites of rainwater discharge or those near anthropogenic activities. A predominance of the dinoflagellate Pyrodinium bahamense was found with abundances of up to 10⁴ cells L⁻¹. Identification of the dinoflagellate was corroborated by light and scanning electron microscopy. Samples for toxins were positive by IFL, and the analogs NeoSTX and STX were identified and quantified by HPLC-FLD and UHPLC-MS/MS, with a total PST concentration of 6.5 pg cell⁻¹. This study is the first report that confirms the presence of PSTs in P. bahamense in Mexican waters of the Gulf of Mexico.

Allelopathy between phytoplankton organisms is promoted by substances released into the marine environment that limit the presence of the dominating species. We evaluated the allelopathic effects and response of cell-free media of Chattonella marina var. marina and Gymnodinium impudicum in the toxic dinoflagellate Gymnodinium catenatum. Additionally, single- and four-cell chains of G. catenatum isolated from media with allelochemicals were cultured to evaluate the effects of post exposure on growth and cell viability. Cell diagnosis showed growth limitation and an increase in cell volume, which reduced mobility and led to cell lysis. When G. catenatum was exposed to cell-free media of C. marina and G. impudicum, temporary cysts and an increased concentration of paralytic shellfish toxins were observed. After exposure to allelochemicals, the toxin profile of G. catenatum cells in the allelopathy experiments was composed of gonyautoxins 2/3 (GTX2/3), decarcarbamoyl (dcSTX, dcGTX2/3), and the sulfocarbamoyl toxins (B1 and C1/2). A difference in toxicity (pg STXeq cell⁻¹) was observed between G. catenatum cells in the control and those exposed to the filtrates of C. marina var. marina and G. impudicum. Single cells of G. catenatum had a lower growth rate, whereas chain-forming cells had a higher growth rate. We suggest that a low number of G. catenatum cells can survive the allelopathic effect. We hypothesize that the survival strategy of G. catenatum is migration through the chemical cloud, encystment, and increased toxicity.

The harmful microalgae Gymnodinium catenatum is a unique naked dinoflagellate that produces paralytic shellfish poisoning toxins (PSTs). This species is common along the coasts of the Mexican Pacific and is responsible for paralytic shellfish poisoning, which has resulted in notable financial losses in both fisheries and aquaculture. In the Gulf of California, G. catenatum has been related to mass mortality events in fish, shrimp, seabirds, and marine mammals. In this study, the growth, toxin profiles, and toxin content of four G. catenatum strains isolated from Bahía de La Paz (BAPAZ) and Bahía de Mazatlán (BAMAZ) were evaluated with different N:P ratios, keeping the phosphorus concentration constant. All strains were cultivated in semi-continuous cultures (200 mL, 21.0 °C, 120 µmol photon m⁻²s⁻¹, and a 12:12 h light-dark cycle) with f/2 + Se medium using N:P ratios of: 4:1, 8:1, 16:1, 32:1, and 64:1. Paralytic toxins were analyzed by HPLC with fluorescence detection. Maximum cellular abundance and growth were obtained at an N:P ratio of 64:1 (3188 cells mL⁻¹ and 0.34 div day⁻¹) with the BAMAZ and BAPAZ strains. A total of ten saxitoxin analogs dominated by N-sulfocarbamoyl (60–90 mol%), decarbamoyl (10–20 mol%), and carbamoyl (5–10 mol%) toxins were detected. The different N:P ratios did not cause significant changes in the PST content or toxin profiles of the strains from both bays, although they did affect cell abundance.

Most of the shellfish fisheries of Mexico occur in the Gulf of California. In this region, known for its high primary productivity, blooms of diatoms and dinoflagellates are common, occurring mainly during upwelling events. Dinoflagellates that produce lipophilic toxins are present, where some outbreaks related to okadaic acid and dinophisystoxins have been recorded. From January 2015 to November 2017 samples of three species of wild bivalve mollusks were collected monthly in five sites in the southern region of Bahía de La Paz. Pooled tissue extracts were analyzed using LC-MS/MS to detect lipophilic toxins. Eighteen analogs of seven toxin groups, including cyclic imines were identified, fortunately individual toxins did not exceed regulatory levels and also the total toxin concentration for each bivalve species was lower than the maximum permitted level for human consumption. Interspecific differences in toxin number and concentration were observed in three species of bivalves even when the samples were collected at the same site. Okadaic acid was detected in low concentrations, while yessotoxins and gymnodimines had the highest concentrations in bivalve tissues. Although in low quantities, the presence of cyclic imines and other lipophilic toxins in bivalves from the southern Gulf of California was constant.

Microcystins (MCs) produced by cyanobacteria are a ubiquitous worldwide problem because some MCs can cause tumor formation and are hepatotoxic. In the Santa María del Oro crater lake, Mexico, plankton scums are recurrent during most of the year and are associated with cyanobacteria of the genera Microcystis spp. and Lyngbya spp. As some of these species are associated with the production of MCs and paralytic shellfish toxins (PSTs), samples from these scums and particulate matter were collected and analyzed for the main bloom species and toxins by a ultrahigh performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) and high performance liquid chromatography with fluorescence detection (HPLC-FLD). Results showed that the main bloom-forming species were Limnoraphis robusta and Microcystis aeruginosa, the presence of at least seven MC congeners and the absence of PSTs in the algae scums. The MCs identified were MC-WR, MC-LR, MC-LA, MC-HilR, MC-LF, MC-YR, and MC-LY. On a dry mass weight basis, MC concentrations were low and ranged between 0.15 and 6.84 μg/kg. Toxin profiles were dominated by MC-WR, MC-LR, and MC-LA, representing 94.5% of the total sample, with each analog contributing 39.8%, 38.1% and 16.5% by relative concentration, respectively. Two of the more hazardous congeners, MC-LR and MC-LA, represented 54.6% of the total MC concentration. MCs in particulate matter along the depth profile were not detected. The MC profile is linked to M. aeruginosa, and it represents the first quantitative MC congener description for this species from a Mexican water ecosystem. Since these mats are recurrent yearly, their effects on humans and wild fauna, and the possible role of anthropogenic activities that favor their presence and proliferation, need to be evaluated.

Historical records of ciguatera in Mexico date back to 1862. This review, including references and epidemiological reports, documents 464 cases during 25 events from 1984 to 2013: 240 (51.72%) in Baja California Sur, 163 (35.12%) in Quintana Roo, 45 (9.69%) in Yucatan, and 16 (3.44%) cases of Mexican tourists intoxicated in Cuba. Carnivorous fish, such as snapper (Lutjanus) and grouper (Epinephelus and Mycteroperca) in the Pacific Ocean, and great barracuda (Sphyraena barracuda) and snapper (Lutjanus) in the Atlantic (Gulf of Mexico and Caribbean Sea), were involved in all cases. In the Mexican Caribbean, a sub-record of ciguatera cases that occurred before 1984 exists. However, the number of intoxications has increased in recent years, and this food poisoning is poorly studied in the region. Current records suggest that ciguatera fish poisoning in humans is the second most prevalent form of seafood poisoning in Mexico, only exceeded by paralytic shellfish poisoning (505 cases, 21 fatalities in the same 34-year period). In this study, the status of ciguatera in Mexico (epidemiological and treatment), and the fish vectors are reviewed. Dinoflagellate species Gambierdiscus, Ostreopsis, and Prorocentrum are related with the reported outbreaks, marine toxins, ecological risk, and the potential toxicological impact.

Saxitoxin (STX) and its analogs are paralytic alkaloid neurotoxins that block the voltage-gated sodium channel pore (Na_v), impeding passage of Na⁺ ions into the intracellular space, and thereby preventing the action potential in the peripheral nervous system and skeletal muscle. The marine dinoflagellate Gymnodinium catenatum produces an array of such toxins, including the recently discovered benzoyl analogs, for which the mammalian toxicities are essentially unknown. We subjected STX and its analogs to a theoretical docking simulation based upon two alternative tri-dimensional models of the Na_v1.4 to find a relationship between the binding properties and the known mammalian toxicity of selected STX analogs. We inferred hypothetical toxicities for the benzoyl analogs from the modeled values. We demonstrate that these toxins exhibit different binding modes with similar free binding energies and that these alternative binding modes are equally probable. We propose that the principal binding that governs ligand recognition is mediated by electrostatic interactions. Our simulation constitutes the first in silico modeling study on benzoyl-type paralytic toxins and provides an approach towards a better understanding of the mode of action of STX and its analogs.