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.     

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.     

Evaluation of the analgesic effect of fentanyl–ketamine and fentanyl–lidocaine constant rate infusions in isoflurane-anesthetized dogs undergoing thoracolumbar hemilaminectomy

ObjectiveTo evaluate anesthetic conditions and postoperative analgesia with the use of intraoperative constant rate infusions (CRIs) of fentanyl–lidocaine or fentanyl–ketamine in dogs undergoing thoracolumbar hemilaminectomy.Study designProspective, randomized, blinded, clinical study.AnimalsA total of 32 client-owned dogs.MethodsDogs were premedicated with fentanyl (5 μg kg^–1) administered intravenously (IV), anesthesia was induced with IV alfaxalone and maintained with isoflurane. Fentanyl (0.083 μg kg^–1 minute^–1) was infused IV with either ketamine (0.5 mg kg^–1; then 40 μg kg^–1 minute^–1; group KF) or lidocaine (2 mg kg^–1; then 200 μg kg^–1 minute^–1; group LF) assigned randomly. Heart rate, noninvasive arterial pressures, respiratory rate, esophageal temperature, end-tidal partial pressure of carbon dioxide and isoflurane concentration were recorded throughout anesthesia. Maintenance of anesthesia, recovery and postoperative pain (Glasgow Composite Pain Scale) were scored. Cardiopulmonary data were analyzed using a two-way anova with repeated measures, demographics of the two groups with a t test, and scores with Mann–Whitney U test, with p < 0.05.ResultsAll dogs recovered from anesthesia without complications. No significant difference was found between groups for cardiopulmonary variables, total anesthesia time, sedation score and requirement for postoperative sedation or for rescue analgesia. Anesthetic maintenance score was of lower quality in KF than in LF [median (interquartile range): 0 (0–0.5) versus 0 (0–0); p = 0.032)], but still considered ideal. Recovery score was higher and indicative of less sedation in LF than in KF [1 (1–1.5) versus 0.5 (0–1); p < 0.0001]. Pain score was higher in KF than in LF [2 (1–3) versus 1 (1–2); p = 0.0009].Conclusions and clinical relevanceBoth CRIs of KF and LF provided adequate anesthetic conditions in dogs undergoing thoracolumbar hemilaminectomy. Based on requirement for rescue analgesia, postoperative analgesia was adequate in both groups.     

Earthworm populations and growth rates related to long-term crop residue and tillage management

Conventional tillage creates soil physical conditions that may restrict earthworm movement and accelerate crop residue decomposition, thus reducing the food supply for earthworms. These negative impacts may be alleviated by retaining crop residues in agroecosystems. The objective of this study was to determine the effects of various tillage and crop residue management practices on earthworm populations in the field and earthworm growth under controlled conditions. Population assessments were conducted at two long-term (15+ years) experimental sites in Québec, Canada with three tillage systems: moldboard plow/disk harrow (CT), chisel plow or disk harrow (RT) and no tillage (NT), as well as two levels of crop residue inputs (high and low). Earthworm growth was assessed in intact soil cores from both sites. In the field, earthworm populations and biomass were greater with long-term NT than CT and RT practices, but not affected by crop residue management. Laboratory growth rates of Aporrectodea turgida (Eisen) in intact soil cores were affected by tillage and residue inputs, and were positively correlated with the soil organic C pool, suggesting that tillage and residue management practices that increase the soil organic C pool provide more organic substrates for earthworm growth. The highest earthworm growth rates were in soils from RT plots with high residue input, which differed from the response of earthworm populations to tillage and residue management treatments in the field. Our results suggest that tillage-induced disturbance probably has a greater impact than food availability on earthworm populations in cool, humid agroecosystems.     

Irrigation scheduling of plastic greenhouse vegetable crops based on historical weather data

Irrigation scheduling based on the daily historical crop evapotranspiration (ET_h) data was theoretically and experimentally assessed for the major soil-grown greenhouse horticultural crops on the Almería coast in order to improve irrigation efficiency. Overall, the simulated seasonal ET_h values for different crop cycles from 41 greenhouses were not significantly different from the corresponding values of real-time crop evapotranspiration (ET_c). Additionally, for the main greenhouse crops on the Almería coast, the simulated values of the maximum cumulative soil water deficit in each of the 15 consecutive growth cycles (1988–2002) were determined using simple soil-water balances comparing daily ET_h and ET_c values to schedule irrigation. In most cases, no soil-water deficits affecting greenhouse crop productivity were detected, but the few cases found led us to also assess experimentally the use of ET_h for irrigation scheduling of greenhouse horticultural crops. The response of five greenhouse crops to water applications scheduled with daily estimates of ET_h and ET_c was evaluated in a typical enarenado soil. In tomato, fruit yield did not differ statistically between irrigation treatments, but the spring green bean irrigated using the ET_h data presented lower yield than that irrigated using the ET_c data. In the remaining experiments, the irrigation-management method based on ET_h data was modified to consider the standard deviation of the inter-annual greenhouse reference ET. No differences between irrigation treatments were found for productivity of pepper, zucchini and melon crops.