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.     

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.     

Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco

Cotyledons from dissected sterile embryos of salgareño pine (Pinus nigra Arn. ssp. salzmannii (Dunal) Franco) were inoculated with different disarmed Agrobacterium tumefaciens strains harbouring the binary vector p35SGUSint. The transient expression of a β-glucuronidase gene (uidA) was studied, using a histochemical staining procedure. Nineteen days after inoculation, the activity of β-glucuronidase was detected in epidermal and subepidermal layers of cotyledonary explants. The EHA105 strain harbouring a disarmed agropine-type Ti-plasmid (pTiBO542) was the most effective for gene transfer of the uidA gene. The effects of exudates and extracts from 0-day-old embryos on induction of vir gene expression in A. tumefaciens were also examined. The results of this study showed that salgarño pine embryo exudates contain a substance(s) that induce vir gene expression, in similar way to that observed with 100 μM acetosyringone (AS).All these findings suggest that T-DNA processing and transfer might take place when Agrobacterium infects suitable tissues of salgareño pine.     

Sclerotinia resistance in sunflower: I. Genotypic variations of hybrids in three environments of Argentina

Sclerotinia sclerotiorum infections on sunflower capitula produce white rot, one of the most dangerous diseases sunflower bears in all humid areas. Therefore, disease quantification on developed genetic materials is a very important action in sunflower breeding. Given the horizontal type of resistance these evaluations should be made in different environments. Several hybrids obtained after crossing seven female × seven male sunflower inbred lines in a factorial mating design were evaluated for resistance to white rot in two locations in the southeast of the province of Buenos Aires, Argentina. Disease incidence and the symptom appearance index indicated significant genotypic effects and genotype × environment (G × E) interactions. Genotypic effects were a greater source of variance than the G × E interaction effects. The G × E interactions only represented changes in magnitude. These results were used to identify the best genotypes for both resistance factors. Four hybrids showed low disease incidence values under both artificial and natural infections, of which only two had high values of the symptom appearance index at both locations. These hybrids are considered to have adequate resistance attributes for the ascosporic penetration and the mycelium extension in the capitulum tissue and could therefore be sown in the province of Buenos Aires without increasing risks of S. sclerotiorum attacks.     

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.     

Intimal thickening of coronary arteries in the rabbitfish,Chimaera monstrosa L. (Chondrichthyes: Holocephali)

This study is the first to report the occurrence of intimal thickening of coronary arteries in a holocephalan, namely the rabbitfish, Chimaera monstrosa. The sample studied consisted of five hearts from rabbitfish with body weights ranging between 12 g and 1116 g. The specimens were fixed in formalin, in methanol:acetone:water (MAW), or in paraformaldehyde and were examined by brightfield and polarization light microscopy. Coronary arteries from three larger animals displayed focal intimal thickenings, which were located in the left main coronary artery trunk, at the level of the cono‐ventricular junction, and in several intramyocardial ventricular arteries. The vascular changes were characterized by myointimal proliferation, breakage or absence of the inner elastic lamina, and, in one case, by increased collagen within the myointimal proliferation. Taking into account the severity of the coronary arterial changes, their location, and the body weight of the affected animals, we conclude that (1) these changes are age‐related in the rabbitfish and (2) local intense mechanical tension seems to be an important factor in their formation in this species.     

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.     

Total chemically available (free and intrachain) lysine and furosine in pea, bean, and lentil sprouts

The effect of the germination of peas, beans, and lentils under differing conditions of illumination for different times on parameters linked to the Maillard reaction (chemically available free and intrachain lysine, lysine availability, and furosine) was evaluated. The chemically available free lysine content in the raw seeds of the three legumes was quite small compared to the chemically available intrachain lysine content, and furosine was detectable only in the beans and the lentils. The effect of germination was to increase lysine availability compared with levels in the raw seeds in all of the germinated samples, the smallest increase taking place in the lentils. In addition, furosine became detectable in all of the germinated samples. Quantities varied depending on the germination conditions but in all cases were higher than the quantities observed in the raw seeds. Linear correlations were observed to exist between some of the parameters considered in the three legumes tested.