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.     

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.     

Efficacy of bovine herpesvirus-1 inactivated vaccine against abortion and stillbirth in pregnant heifers

OBJECTIVE: To evaluate the efficacy of an inactivated bovine herpesvirus-1 (BHV-1) vaccine to protect against BHV-1 challenge-induced abortion and stillbirth. DESIGN: Prospective study. ANIMALS: 35 beef heifers. PROCEDURES: Before breeding, heifers were vaccinated with a commercially available BHV-1 inactivated vaccine SC or IM. The estrous cycle was then synchronized, and heifers were artificially inseminated 30 to 60 days after vaccination. Heifers (n = 21) were challenge inoculated IV at approximately 180 days of gestation with virulent BHV-1. Fourteen control heifers were not vaccinated. Clinical signs of BHV-1 infection were monitored for 10 days following challenge; serologic status and occurrence of abortion or stillbirth were evaluated until time of calving. RESULTS: 18 of 21 (85.7%) heifers that received vaccine were protected from abortion following challenge, whereas all 14 control heifers aborted. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that an inactivated BHV-1 vaccine can protect against abortion resulting from a substantial challenge infection, with efficacy similar to that of modified-live BHV-1 vaccines.     

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.     

Reduced-calorie Avocado Paste Attenuates Metabolic Factors Associated with a Hypercholesterolemic-high Fructose Diet in Rats

The objective of this study was to evaluate the effect of reduced-calorie avocado paste on lipid serum profile, insulin sensitivity, and hepatic steatosis in rats fed a hypercholesterolemic-high fructose diet. Thirty five male Wistar rats were randomly separated in five groups: Control group (ground commercial diet); hypercholesterolemic diet plus 60 % fructose solution (HHF group); hypercholesterolemic diet plus 60 % fructose solution supplemented with avocado pulp (HHF+A group); hypercholesterolemic diet plus 60 % fructose solution supplemented with reduced-calorie avocado paste (HHF+P group); and hypercholesterolemic diet plus 60 % fructose solution supplemented with a reduced-calorie avocado paste plus fiber (HHF+FP group). The A, P, and FP were supplemented at 2 g/kg/d. The study was carried out for seven weeks. Rats belonging to the HHF group exhibited significantly (P?≤?0.05) higher total cholesterol, triglycerides, and insulin levels in serum as well as lower insulin sensitivity than the control group. Supplementation with reduced-calorie avocado paste showed a significant (P?≤?0.05) decrease in total cholesterol (43.1 %), low-density lipoprotein (45.4 %), and triglycerides (32.8 %) in plasma as well as elevated insulin sensitivity compared to the HHF group. Additionally, the liver enzymes alanine aminotransferase and aspartate aminotransferase decreased significantly in the HHF-P group (39.8 and 35.1 %, respectively). These results are likely due to biocompounds present in the reduced-calorie avocado paste, such as polyphenols, carotenoids, chlorophylls, and dietary fibre, which are capable of reducing oxidative stress. Therefore, reduced-calorie avocado paste attenuates the effects of a hypercholesterolemic-high fructose diet in rats.     

pH-Dependent Solution Structure and Activity of a Reduced Form of the Host-Defense Peptide Myticin C (Myt C) from the Mussel Mytilus galloprovincialis

Myticin C (Myt C) is a highly variable host-defense peptide (HDP) associated to the immune response in the mediterranean mussel (Mytilus galloprovincialis), which has shown to be active across species due to its strong antiviral activity against a fish rhabdovirus found in fish cells overexpressing this HDP. However, the potential antimicrobial properties of any synthetic analogue of Myt C has not yet been analysed. Thus, in this work we have synthesised the sequence of the mature peptide of Myt C variant c and analysed the structure activity relationships of its reduced (non-oxidized) form (red-MytCc). In contrast to results previously reported for oxidized isoforms of mussel myticins, red-MytCc was not active against bacteria at physiological pH and showed a moderate antiviral activity against the viral haemorrhagic septicaemia (VHS) rhabdovirus. However, its chemotactic properties remained active. Structure/function studies in neutral and acid environments by means of infrared spectroscopy indicated that the structure of red-MytCc is pH dependent, with acid media increasing its alpha-helical content. Furthermore, red-MytCc was able to efficiently aggregate artificial phospholipid membranes at low pH, as well as to inhibit the Escherichia coli growth, suggesting that this activity is attributable to its more structured form in an acidic environment. All together, these results highlight the dynamic and environmentally sensitive behavior of red-Myt C in solution, and provide important insights into Myt C structure/activity relationships and the requirements to exert its antimicrobial/immunomodulatory activities. On the other hand, the pH-dependent direct antimicrobial activity of Myt C suggests that this HDP may be a suitable template for the development of antimicrobial agents that would function selectively in specific pH environments, which are sorely needed in this “antibiotic-resistance era”.