Stability of Saxitoxin in 50% Methanol Fecal Extracts and Raw Feces from Bowhead Whales (Balaena mysticetus)

In recent decades, harmful algal blooms (HABs) producing paralytic shellfish toxins (including saxitoxin, STX) have become increasingly frequent in the marine waters of Alaska, USA, subjecting Pacific Arctic and subarctic communities and wildlife to increased toxin exposure risks. Research on the risks of HAB toxin exposures to marine mammal health commonly relies on the sampling of marine mammal gastrointestinal (GI) contents to quantify HAB toxins, yet no studies have been published testing the stability of STX in marine mammal GI matrices. An understanding of STX stability in test matrices under storage and handling conditions is imperative to the integrity of toxin quantifications and conclusions drawn thereby. Here, STX stability is characterized in field-collected bowhead whale feces (stored raw in several treatments) and in fecal extracts (50% methanol, MeOH) over multiple time points. Toxin stability, as the percent of initial concentration (T0), was reported for each storage treatment and time point. STX was stable (mean 99% T0) in 50% MeOH extracts over the 8-week study period, and there was no significant difference in STX concentrations quantified in split fecal samples extracted in 80% ethanol (EtOH) and 50% MeOH. STX was also relatively stable in raw fecal material stored in the freezer (mean 94% T0) and the refrigerator (mean 93% T0) up to 8 weeks. STX degraded over time in the room-temperature dark, room-temperature light, and warm treatments to means of 48 ± 1.9, 38 ± 2.8, and 20 ± 0.7% T0, respectively, after 8 weeks (mean ± standard error; SE). Additional opportunistically analyzed samples frozen for ≤4.5 years also showed STX to be relatively stable (mean 97% T0). Mean percent of T0 was measured slightly above 100% in some extracts following some treatments, and (most notably) at some long-term frozen time points, likely due to evaporation from samples causing STX to concentrate, or variability between ELISA plates. Overall, these results suggest that long-term frozen storage of raw fecal samples and the analysis of extracts within 8 weeks of extraction in 50% MeOH is sufficient for obtaining accurate STX quantifications in marine mammal fecal material without concerns about significant degradation.     

Phenolic acids in foods: an overview of analytical methodology

Phenolic acids are aromatic secondary plant metabolites, widely spread throughout the plant kingdom. Existing analytical methods for phenolic acids originated from interest in their biological roles as secondary metabolites and from their roles in food quality and their organoleptic properties. Recent interest in phenolic acids stems from their potential protective role, through ingestion of fruits and vegetables, against oxidative damage diseases (coronary heart disease, stroke, and cancers). High performance liquid chromatography (HPLC) as well as gas chromatography (GC) are the two separation techniques reviewed. Extraction from plant matrixes and cleavage reactions through hydrolysis (acidic, basic, and enzymatic) are discussed as are the derivatization reagents used in sample preparation for GC. Detection systems discussed include UV-Vis spectroscopy, mass spectrometry, electrochemical, and fluorometric detection. The most common tandem techniques are HPLC/UV and GC/MS, yet LC/MS is becoming more common. The masses and MS fragmentation patterns of phenolic acids are discussed and tabulated as are the UV absorption maxima.     

Organophosphorus pesticide residues in Mexican commercial pasteurized milk

A study was conducted to measure residues of 13 organophosphorus (OP) pesticides, widely used as dairy cattle ectoparasiticides or in crops used for animal feed, in homogenized and pasteurized Mexican milk samples. Four different milk brands with high distribution were collected biweekly during a 12 month period (n = 96) in supermarkets. OP pesticide residues were measured by gas chromatography with a flame photometric detector. Approximately 39.6% of the samples contained detectable levels of OP pesticide residues. Eight samples contained residues exceeding established maximum residue limits (MRL), and the OP pesticides present in these samples were dichlorvos (five samples), phorate, chlorpyrifos, and chlorfenvinphos (one sample, respectively). Average residues of 13 OP pesticides measured were below established MRLs ranging between 0.0051 and 0.0203 ppm.     

Modified Pollard transects for assessing tropical butterfly abundance and diversity

This paper introduces and discusses the consistency and effectiveness of an inexpensive modification of Pollard transects for assessing the diversity and abundance of tropical butterfly communities in two national parks in Rio de Janeiro, Brazil. To measure butterfly abundance, students walked simultaneous timed transects at the interface of forest and cleared areas. They either counted the number of individuals observed or tabulated the number of specimens collected with nets. After a short training period, the number of butterflies observed or collected on simultaneous transects was statistically indistinguishable among student groups, and there was a significant positive correlation between observation counts and collected number of individuals. As a measure of species richness, the number of butterfly species sampled on each simultaneous transect was tabulated and did not differ statistically. To measure diversity, alpha of the logseries model was calculated for each collected sample, and statistical fit to a logseries model was determined. Although virtually all daily samples and the year's accumulated sample at one park fit the logseries model, about 35% of the daily samples and the entire year's sample did not fit at the other park. Despite these differences between the two parks, values for alpha from daily samples at both parks varied similarly (from 15 to 50 in almost all cases), and values from the entire year's samples were statistically indistinguishable. The repeatability of results among novices, such as students, suggests that timed transects have great promise for furthering our understanding of butterfly community demographics.     

Antihyperlipidemic Effect of Methanolic Extract from Opuntia joconostle Seeds in Mice Fed a Hypercholesterolemic Diet

The purpose of this study was to evaluate the hypolipidemic effect of a methanolic extract from Opuntia joconostle seeds fed to mice in a hypercholesterolemic diet. Acute toxicity of the methanolic extract was investigated by an established method. Phenolic composition and antioxidant activity were determined by high-performance liquid chromatography and DPPH, respectively. The total phenolic content of Opuntia joconostle seeds was 47.85?±?1.29 mg gallic acid equivalents/g dry weight. The main phenolic compounds were identified as quercetin, rutin, and cafeic acid. Percent inhibition of DPPH⁺ was 49.76?±?0.49 %. The oral LD₅₀ for the methanolic extract from the Opuntia joconostle seeds was >5,000 mg/kg BW. Mice fed a hypercholesterolemic diet for six days exhibited significantly (P?≤?0.001) higher plasma lipid levels than mice fed a normal diet. Remarkably, supplementation with methanolic extract from Opuntia joconostle at doses of 1, 2, and 5 g/kg body weight significantly (P?≤?0.001) prevented the increase in total cholesterol, low-density lipoprotein cholesterol, triglycerides level, and atherogenic index. Similar concentrations of the HDL cholesterol were observed in both treated and control groups. A significant dose-dependent reduction in lipid levels was noted for treated groups compared to the hypercholesterolemic group. We attribute this result to the seeds’ phenolic composition. This methanolic extract has potential to be included in short-term hypercholesterolemia treatment regimens as it exhibits hypolipidemic activity with no apparent toxic manifestations.     

Two new species of Clestobothrium (Cestoda: Bothriocephalidea), parasites of Merluccius australis and m. hubbsi (Gadiformes: Merlucciidae) from the Patagonian shelf of Argentina, with comments on Clestobothrium crassiceps (Rudolphi, 1819)

Abstract: Two new species of bothriocephalidean cestodes, Clestobothrium splendidum sp. n. from Merluccius australis (Hutton) and Clestobothrium cristinae sp. n. from Merluccius hubbsi Marini from the Patagonian shelf of Argentina, are described. Clestobothrium splendidum can be typified by the following characteristics: a medium-sized strobila composed of410-528 proglottides that are much wider than long; 49-90 testes per mature proglottis, partially surrounding the ovary posteriorly; a transversely elongated genital pore situated anterior to spurious articulations; presence of a genital atrium; a globular cirrus-sac occupying 4-6% of mature proglottis width; a vagina with sphincter and three pairs of osmoregulatory canals on each side of the proglottis. Clestobothrium cristinae is characterised by its small size; 71-219 proglottides; 39-64 testes per mature proglottis, usually surrounding completely the ovary posteriorly; a rounded genital pore situated at the same level of spurious articulations; an oval cirrus-sac occupying 8-16% of mature proglottis width; and three pairs of osmoregulatory canals on each side of the proglottis. Clestobothrium cristinae shares with C. splendidum the type and distribution of microtriches, except for the central surface delimited by two lips. Additionally, type and voucher materials of Clestobothrium crassiceps (Rudolphi, 1819) from Merluccius merluccius were studied. A key to species is provided.