Viability and nematophagous activity of the freeze-dried fungus Arthrobotrys robusta against Ancylostoma spp. infective larvae in dogs

Viability and in vitro and in vivo activities of freeze-dried conidia of the predatory fungus Arthrobotrys robusta (I-31) were evaluated against infective larvae (L(3)) of Ancylostoma spp. in dogs. A. robusta conidia were lyophilized and stored at 4°C for a month. Freeze-dried conidia were diluted to 1×10(3)conidia/ml and tested in vivo. The treated group consisted of a solution containing conidia (1ml) and 1000 Ancylostoma spp. (L(3)) placed on Petri dishes plated with 2% water-agar (2% WA), at 25°C, in the dark for 10 days. The control group consisted of 1000 Ancylostoma spp. L(3), plated on 2% WA. After 10 days, Ancylostoma spp. L(3) from both the treated and the control groups were recovered and counted. The in vivo test was performed on two dogs by administering a single oral dose of freeze-dried conidia (1.5×10(5)) in aqueous solution to one animal and only water to the other. Fecal samples were collected at 12, 24 and 48h after the treatments, plated 2% WA plates and incubated at 25°C for 15 days. A thousand Ancylostoma spp. L(3) larvae were spread on these plates. At day 15, infective L(3) recovered from the treated and control groups were counted. In the in vitro test, A. robusta was able to survive the freeze-drying process, grow in the plates, form traps and capture Ancylostoma spp. L(3). There was a 75.38% decrease in the number of infective larvae recovered from the treated group. The in vivo test showed that freeze-dried A. robusta conidia survived the passage through the gastrointestinal tract of the treated dog, was able to grow in the plates and capture Ancylostoma spp. L(3), reducing the number of recovered L(3) (p<0.01). Freeze-drying can be an alternative method for conservation of conidia of nematophagous fungi.     

Performance and ruminal protozoa in lambs with chromium supplementation

The effects of chromium picolinate (CrPic) oral supplementation on the performance and ruminal protozoa population in sheep was investigated. Twenty-four male lambs were treated with four different levels of CrPic: placebo, 0.250, 0.375 and 0.500 mg of chromium/animal/day during 84 days. The base ration was Panicum maximum cv Massai hay and concentrate. Feed intake was measured three times a week. Lambs were weighed every 2 weeks. Ruminal content was sampled five times during the trial to quantify ruminal protozoa. No difference (p > 0.05) between treatments was recorded for any parameter measured: initial and final mean body weight, dry matter intake, daily gain and total body weight gain. There was a negative linear relationship between Cr supplementation and protozoa count (p = 0.0013) with no additional decrease when CrPic supplemental levels were higher than 0.375 mg/day. Therefore, Cr supplementation must be carried out carefully and more studies need to address stressed animals or possible toxic effects of Cr in the animal itself or to the ruminal protozoa population.     

Continuous infusion of propofol in dogs premedicated with methotrimeprazine

Objective To evaluate the cardiopulmonary and clinical effects of three different infusion rates of propofol in dogs premedicated with methotrimeprazine. Study design Randomized experimental trial. Animals Ten healthy adult mixed‐breed male and female dogs, weighing from 14 to 20 kg. Methods Dogs were premedicated with methotrimeprazine [1 mg kg^?1 intravenously (IV)] followed by induction of anesthesia with 4.5 mg kg^?1 of propofol IV and maintenance with propofol for 60 minutes as follows: T1, 0.2 mg kg^?1 minute^?1; T2, 0.3 mg kg^?1minute^?1; and T3, 0.4 mg kg^?1minute^?1. Heart rate (HR), respiratory rate (RR), mean arterial pressure (MAP), end‐tidal CO₂ (P_ETCO₂), arterial hemoglobin O₂ saturation, arterial blood gases, and pedal and cutaneous reflexes were measured before and 5, 10, 20, 30, 45 and 60 minutes after the beginning of the propofol infusion. Statistical analysis was performed using an anova . Results Heart rate increased during anesthesia in all cases and arterial blood pressure decreased only in dogs in the T3 category. Respiratory depression was proportional to the infusion rate of propofol. Muscle relaxation was satisfactory, but analgesia was inadequate in the three treatments. Conclusions The infusion of 0.2–0.4 mg kg^?1 minute^?1 of propofol produced a dose‐dependent respiratory depression. The presence of a pedal withdrawal reflex and marked cardiovascular responses to this noxious stimulus suggests that anesthesia may not be of sufficient depth for surgery to be carried out. Clinical relevance Although several studies have been performed using propofol in animals, few studies have investigated the cardiopulmonary and analgesic effects with different doses. The determination of an adequate propofol infusion rate is necessary for the routine use of this intravenous anesthetic for the maintenance of anesthesia during major surgical procedures in dogs.     

Water use efficiency and must quality of irrigated grapevines of north-eastern Portugal

This is an experiment to study the effect of water supply on the quality of the must produced by grapevines, and to determine the water use efficiency (WUE) of different irrigation strategies to give farmers a tool to optimize irrigation, reducing water use with the least negative effect on production. WUE was based on the ratio between a given parameter (yield, different must solubes) and the water used by plant transpiration plus evaporation from soil. The experiment was a completely randomized design with six irrigation treatments (non-irrigated, irrigated from flowering to veraison at rates of 4 and 8 mm day^?1, irrigated from veraison to commercial maturation at rates of 4 and 8 mm day^?1, and irrigated from flowering to commercial maturation at a rate of 8 mm day^?1) in three replications over three seasons (2004–2006). Irrigation significantly increased grapevine yield and, because the plants produced larger berries, there was a dilution of colour, aroma and soluble solids that corresponded to a lower quality of the must. The higher WUE was achieved with no irrigation but yield was very low, albeit with an outstanding quality, that might not be profitable. The best strategy is to balance the water stress of the vines that best combines yield and berry quality and this was achieved by irrigating from flowering to veraison at a rate of ～50% of the potential evapotranspiration. This strategy might conserve water, an increasingly scarce resource that we are pressed to use with the utmost efficiency.     

Production of a high percentage of male offspring in growth-enhanced transgenic tilapia using Oreochromis aureus ZZ selected pseudofemales

Gene transfer has offered a new tool for the development of improved fish strains for aquaculture. Monosex fish populations could minimize the impact of genetically modified organisms in the environment. In Oreochromis aureus, the use of pseudofemale spawners (sex-reversed male with a female phenotype) is an alternative technique for producing genetically male tilapia offspring. O. aureus fry were treated with 17β-estradiol at 100 mg/kg of food for 45 days. We obtained 77.1% females and 45.9% in the control group. Females randomly taken from the treated group were crossed with normal males. Fry from pseudofemales producing more than 90% male progenies were submitted to 17 β-estradiol treatment to obtain F₂ pseudofemales. The results of the sex-reversal were low and variable ranging between 66.0 and 84.3% females. F₂ pseudofemales were crossed with transgenic males from the F70 line (O. aureus × O. urolepis hornorum) and non-transgenic (O. aureus) males. The sex ratio of progeny of F₂ pseudofemale deviated significantly (P < 0.01) in favor of males in the crosses with transgenic (90.2%) and non-transgenic (89.3%) males compared to the results observed with normal females (51.0 and 52.3%, respectively). The mean fry production with pseudofemales (per m²/day) was similar to the normal females in the crosses with transgenic and non-transgenic males. To our knowledge this is the first report on the production of a near monosex population in genetically modified fish.     

First feeding of common carp larvae on diets with high levels of protein hydrolysates

A 21 day feeding trial was carried out at 24 °C aiming to evaluate the effect of diets containing high levels of protein hydrolysates on growth, survival and body composition of common carp, Cyprinus carpio L., larvae since first feeding. Eight semipurified diets based on a fish protein hydrolysate (CPSP) with or without additional nitrogen sources, such as casein, casein hydrolysates or other hydrolysates from several origins, were tested, using a diet based on yeast as a reference. High survival rates of larvae were observed at the end of the trial for all experimental diets, ranging from 97% for the yeast diet to 71% when CPSP was used as the only nitrogen source. In terms of growth, the best results were achieved with the CPSP-casein diet (1:1) and the yeast-based diet (17.5 mm total length / 82.8 mg wet weight and 16.3 mm total length / 55.4 mg wet weight, respectively). Growth of larvae fed on diets with protein hydrolysates as the only nitrogen source was considerably lower than that of larvae fed a mixture of intact protein and protein hydrolysates.     

Electrocardiographic evaluation of the degree of sedation and the isolated use of methadone in healthy dogs

ObjectiveThe present study aimed to investigate the influence of methadone on cardiorespiratory parameters, electrocardiogram and clinical sedation in dogs. Further possible side effects are reported.Study designProspective experimental cross-over study.DogsEight, 1–4-year-old, various breeds of dogs of both genders weighing 9–36 kg.MethodsEach dog was treated three times: methadone 0.3 mg kg^?1 (M0.3), 0.5 mg kg^?1 (M0.5) and 1.0 mg kg^?1 (M1.0) intramuscularly. Respiratory rate, heart rate and arterial blood pressure were recorded as well as electrocardiographic evaluation of lead II. Clinical sedation in each treatment received a score (0–3) after drug administration and at 30 minute intervals until scores and measurements returned to baseline values.ResultsA significant decrease in heart rate was seen with each dose of methadone and bradycardia (HR<60 bpm) was noted in a few dogs at each dose. A clinically significant arrhythmia occurred in one dog at 1 mg kg^?1 that required reversal with butorphanol. There was no significant difference in SAP, MAP and DAP between treatments. Some side effects such as salivation, defecation, vocalization and panting, after administration of methadone were observed. There were no differences in mean values of heart rate, P-wave and QRS complex duration and QT interval between treatments.Conclusion and clinical relevanceMethadone administration was associated with panting and a decrease in heart rate at all doses tested in this study. The cardiac rhythm should be monitored carefully in dogs when methadone is administered on its own, especially at higher doses.     

Tumescent local anesthesia with ropivacaine in different concentrations in bitches undergoing mastectomy: plasma concentration and post‐operative analgesia

ObjectiveTo compare two concentrations of ropivacaine administered for tumescent local anesthesia (TLA) in dogs undergoing mastectomy.Study designProspective randomized clinical study.AnimalsSeventeen bitches of various breeds, aged 12 ± 2 years and weighing 10 ± 6.5 kg requiring total unilateral or bilateral mastectomy.MethodsDogs were premedicated with acepromazine (0.04 mg kg^?1) and morphine (0.4 mg kg^?1) intramuscularly. Anesthesia was induced with propofol (2.5 mg kg^?1) and midazolam (0.2 mg kg^?1) intravenously, followed by intubation and maintenance with isoflurane and TLA. Dogs were randomly allocated to receive TLA either with 0.1% ropivacaine (group G1) or with 0.05% ropivacaine (group G05). TLA was performed by insertion of a multihole needle under the skin and infusion of ropivacaine and lactated Ringer’s solution at a fixed volume of 15 mL kg^?1. Ropivacaine concentrations in arterial blood were measured by high-performance liquid chromatography. Post-operative pain was assessed using two scales (University of Melbourne pain scale and a modified composite measure pain scale) and von Frey filaments, 4 hours after TLA and at 1 hour intervals until sensitivity was regained. A score above 30% of the maximum possible score was considered a positive indicator of pain.ResultsPeak plasma concentrations of ropivacaine were measured 240 minutes after TLA in G1. Low concentrations were measured in G05 for 60 minutes, with subsequent increase. Analgesic rescue and return of sensitivity occurred at 7 ± 2.3 and 7 ± 1.9 hours (mean ± SD) after TLA for G1 and G05, respectively.Conclusions and clinical relevanceTumescent local anesthesia with ropivacaine provided satisfactory post-operative analgesia that lasted for several hours, with no difference in duration between the concentrations. No serious side effects were attributed to TLA. Results indicated that 0.05% ropivacaine provided adequate analgesia for mastectomy, however, more studies are required to support this conclusion.     

Protease inhibitor activity is associated to a basic chitinase from potato but not to an acidic one

Summary Several pathogenesis-related proteins, which are produced in plants submitted to stress, have been identified as chitinases. We have previously described that a potato basic chitinase strongly inhibited the activity of an aspartic protease isolated from the same source. In this work, we have tested the activity of two potato chitinases as protease inhibitors. A basic chitinase (ChiB) inhibited trypsin, chymotrypsin, subtilisin, proteinase K and a serine protease ofFusarium sp. An acidic one (ChiA) did not show inhibitory activity. The kinetics of trypsin inhibition by ChiB revealed different patterns of inhibition with azocasein and BAPNA. Metal ions affected differentially both activities, suggesting that ChiB is a bifunctional protein. These results and those reported by other authors suggest a new physiological role for pathogenesis-related proteins. However, results presented in this paper suggest that the protease inhibitor activity is not a general characteristic of potato chitinases.     

Regulatory Factors and Structure of a Component Population of the Spionid Polydora bioccipitalis Infesting the Surf Clam Mesodesma donacium

Abstract

Spionidae, particularly polydorids, are common polychaete parasites of edible mollusks around the world. However, our understanding of the regulatory factors and population structure of these parasites is scant. In this study involving Polydora bioccipitalis and the surf clam Mesodesma donacium we evaluated (1) the environmental correlates of the prevalence and mean intensity of the infestation, (2) the relationship between the number of egg capsules and juvenile and adult parasites and the time elapsed since infestation, and (3) the spatial patterns of juveniles and adults within the host. Environmental factors showed no significant correlations with prevalence and mean intensity, suggesting that these factors do not act directly as regulators. Rather, storm surges seemingly induced clam stranding, which in turn affected both the prevalence and intensity of the infestation. The numbers of juveniles and egg capsules in blisters were significantly related to the time since infestation, suggesting mechanisms of use and expansion of the space within the host. Juvenile worms showed an aggregated distribution that was probably related to the episodic nature of infestation events, whereas adults exhibited uniform distributions that probably reflect territorial behavior and reproductive strategies.

Received June 7, 2010; accepted April 7, 2011