Protective activity against oocyst shedding in cats vaccinated with crude rhoptry proteins of the Toxoplasma gondii by the intranasal route

This study evaluated a vaccine made from crude rhoptry proteins of Toxoplasma gondii with Quil-A, which was administered to cats by the intranasal route. Eleven short-hair domestic cats were divided into four groups: G1 (n=3) received three doses (200 microg/dose) of the rhoptry vaccine with Quil-A (20 microg); G2 (n=3) received PBS with Quil-A (20 microg); G3 (n=3) and G4 (n=2) received only PBS. Treatments were administered at days 0, 21, and 42 by the intranasal route. Challenge was done to G1, G2, and G3 animals with 600 cysts of the VEG strain on day 51 (challenge day); G4 animals were unchallenged. The anti-T. gondii IgG and IgA antibody levels from sera were measured by indirect enzyme-linked immunosorbent assay (ELISA). At challenge, two animals from G1 revealed antibody levels for both IgG and IgA; oocysts were not detected in feces of these two cats. There were no differences in hematological values between groups throughout the experiment (p>0.10). Preventable fractions were 67% in G1 and 0% in G2 and G3. Comparatively, G1 animals shed 89.3% and 90.8% less oocysts than G3 and G4, respectively. Two out of three cats were protected against T. gondii oocyst shedding when the rhoptry vaccine was administered by the intranasal route. This is the first study using crude rhoptry proteins as vaccine by the intranasal route in cats to evaluate protection against oocysts shedding.     

Isolation and molecular detection of Neospora caninum from naturally infected sheep from Brazil

Neospora caninum was isolated from a naturally infected sheep from Brazil by bioassay in dogs. Approximately 70g of brain from each of two 4-month-old sheep with indirect fluorescent antibodies (>or=1:50) to N. caninum was offered to a different IFAT negative dog (Sheep n. 302, IFAT 1:400-Dog 1 and Sheep n. 342, IFAT 1:50-Dog 2). Parasite DNA was detected in both sheep brains using a PCR targeting the Nc-5 gene of N. caninum. Shedding of Neospora-like oocysts was noticed only in Dog 1, from 10 days post-inoculation (PI) to 25 days PI (a total of approximately 27,600 oocysts). Seventy days after infection, Dog 1 was euthanized and brain/cerebellum and medulla were collected and submitted to molecular methods, as were the oocysts, to confirm the identity of the isolate. Serum samples collected weekly from both dogs from the infection to the end of the experimental period had no antibodies anti-N. caninum by IFAT (<1:50). Oocysts, brain/cerebellum and medulla specimens of Dog 1 proved positive by a PCR assay targeting the Nc-5 gene of N. caninum. In addition, the oocysts have the DNA amplified by a PCR based on primers directed to the common toxoplasmatiid ITS1 sequence. The PCR products of ITS1 were sequenced, confirming again the isolate as N. caninum. Oocysts were also orally inoculated in two Swiss white mice two Mongolian gerbils (Meriones ungulatus) and two large vesper mice (Calomys callosus) (10(3)oocysts/animal). The rodents were sacrificed 2 months PI, and fresh preparations of brains showed Neospora thick-walled cysts in gerbil brains, but molecular detection using the Nc-5 PCR assay revealed DNA parasite in gerbil and also C. callosus brains. This is the first report of isolation and sequencing of N. caninum from a Brazilian sheep and the first report of molecular detection of N. caninum from C. callosus.     

Preparation of Extruded Snacks with Flavored Flour Obtained from the Carcasses of Nile Tilapia: Physicochemical,Sensory, and Microbiological Analysis

This study aimed to develop extruded snacks including flour obtained from Nile tilápia carcasses, and then evaluate the chemical composition, colorimetry, thiobarbituric acid reactive substances, and microbiology. There was a significant increase in the levels of crude protein, ether extract, ash, and minerals, as well as a reduction in carbohydrates as more flour was included in the snacks. Increased levels of flavored flour caused reduction in the brightness of snacks. Oxidative rancidity was not detected in snacks. The inclusion of flour improved the nutritional value of extruded snacks without changing sensory characteristics. Microbiologically, the snacks are suitable for human consumption.     

Environmental impact of razor clam harvesting using salt in Ria Formosa lagoon (Southern Portugal) and subsequent recovery of associated benthic communities

• 1. Razor clams are found in different habitats ranging from sheltered systems (e.g. lagoons and estuaries) to open coasts. They are distributed worldwide and comprise a small number of species of high economic value. Depending on the specific habitat and species exploited, different mechanical and hand‐harvesting techniques are employed. While the environmental effects resulting from mechanized methods have been described by several authors, the impacts caused by traditional hand‐harvesting methods remain unknown. Therefore, a study was undertaken in Ria Formosa lagoon (South of Portugal) addressing the environmental effects resulting from harvesting Solen marginatus with salt.
• 2. No significant impact on the sediment was found; the main effect was an increase in salinity after covering the area with salt, which decreased rapidly with the flood tide and after a few hours had returned to pre‐harvesting levels.
• 3. No effects on benthic communities were observed, with similar fluctuation patterns recorded in control and experimental areas, the observed differences being attributed to the natural variability of benthic populations.
• 4. Based on the results obtained in the present study, a razor clam fishery using salt in intertidal areas can be considered environmentally ‘friendly’.

Copyright © 2008 John Wiley & Sons, Ltd.     

Functional Feeds to Tackle Meagre (Argyrosomus regius) Stress: Physiological Responses under Acute Stressful Handling Conditions

Marine algae are recognised sources of bioactive compounds that have attracted great interest as nutritional supplements for aquaculture fish. Intensive rearing conditions often expose fish to husbandry-related stressors, rendering fish more susceptible to disease and reducing production yields. The present work evaluated the potential of two marine algae extracts (Fucus vesiculosus and Nannochloropsis gaditana) as nutritional supplements to mitigate stress effects in meagre (Argyrosomus regius) exposed to an acute handling stress (AS). A plant-based diet was used as a control, and three other diets were prepared, which were similar to the control diet but supplemented with 1% of each algal extract or a combination of the two extracts (0.5% each). The effects of supplemented diets on stress biomarkers, antioxidant enzyme activities, and immune response were analysed in fish exposed to AS after 4 weeks of feeding. Supplemented diets did not affect growth performance but the inclusion of F. vesiculosus promoted higher feed efficiency, as compared to the control group. Dietary algal extracts supplementation reduced plasma glucose levels, increased white blood cell counts, and reduced the expression of pro-inflammatory genes when compared with the control. N. gaditana supplementation led to a reduction in hepatic antioxidant enzyme activity and glutathione levels, while F. vesiculosus supplementation increased muscle glutathione reductase activity and reduced lipid peroxidation. These findings support the potential of algal extracts as nutraceuticals in aquafeeds to enhance the ability of fish to cope with husbandry-related stressful conditions and ultimately improve fish health and welfare.     

From Natural Xanthones to Synthetic C-1 Aminated 3,4-Dioxygenated Xanthones as Optimized Antifouling Agents

Biofouling, which occurs when certain marine species attach and accumulate in artificial submerged structures, represents a serious economic and environmental issue worldwide. The discovery of new non-toxic and eco-friendly antifouling systems to control or prevent biofouling is, therefore, a practical and urgent need. In this work, the antifouling activity of a series of 24 xanthones, with chemical similarities to natural products, was exploited. Nine (1, 2, 4, 6, 8, 16, 19, 21, and 23) of the tested xanthones presented highly significant anti-settlement responses at 50 μM against the settlement of mussel Mytilus galloprovincialis larvae and low toxicity to this macrofouling species. Xanthones 21 and 23 emerged as the most effective larval settlement inhibitors (EC₅₀ = 7.28 and 3.57 µM, respectively). Additionally, xanthone 23 exhibited a therapeutic ratio (LC₅₀/EC₅₀) > 15, as required by the US Navy program attesting its suitability as natural antifouling agents. From the nine tested xanthones, none of the compounds were found to significantly inhibit the growth of the marine biofilm-forming bacterial strains tested. Xanthones 4, 6, 8, 16, 19, 21, and 23 were found to be non-toxic to the marine non-target species Artemia salina (<10% mortality at 50 μM). Insights on the antifouling mode of action of the hit xanthones 21 and 23 suggest that these two compounds affected similar molecular targets and cellular processes in mussel larvae, including that related to mussel adhesion capacity. This work exposes for the first time the relevance of C-1 aminated xanthones with a 3,4-dioxygenated pattern of substitution as new non-toxic products to prevent marine biofouling.     

Effects of an Enterococcus faecium‐based probiotic on growth performance and health of Pirarucu,Arapaima gigas

Intensive fish farming has resulted in an increased concern for disease outbreaks. Probiotic use is one of the strategies being developed to improve fish health and productivity. Measures of probiotic colonization, growth performance, haematological characteristics and parasite load were used to evaluate the effect of diets supplemented with Enterococcus faecium on growth and health of Arapaima gigas juveniles. A completely randomized design with four treatments (diet with E. faecium at 1 × 10⁶ CFU/g and 1 × 10⁸ CFU/g, control diet and diet with the culture medium MRS) and three replicates was used. Ninety‐six Arapaima juveniles were distributed in 12 cages fed with the specified diet for 68 days. Colonization of the intestinal tract by lactic acid bacteria reduced the total number of heterotrophic bacteria in fish fed with probiotics compared to controls. Fish fed a supplemented diet containing 1 × 10⁸ CFU/g presented higher values of weight gain, survival and fish growth uniformity, and lower values of feed conversion ratio. The prevalence of Trichodina sp. could have affected the survival of fish in the control group. Reduction in parasite load and an increase in haematocrit, the number of erythrocytes, thrombocytes, neutrophils and monocytes were also observed in fish fed the diet containing 1 × 10⁸ CFU/g. Enterococcus faecium presented a probiotic effect in A. gigas juveniles and can be recommended for use at a concentration of 1 × 10⁸ CFU/g to modify the gut microbiota, improve growth performance and haematology and reduce parasitic load.     

New organic substrates and boron fertilizing for production of yellow passion fruit seedlings

The identification of alternative substrates suitable for seedling production is very important, while boron nutrition is also relevant due to plant requirement and its difficult management. Thus, four experiments were carried out from October 2012 to April 2013 to evaluate the effect of substrates using decomposed buriti (Mauritia vinifera Mart.) stem (DBS) and boron fertilizing for seedling production of yellow passion fruit (Passiflora edulis Sims). The experimental design was completely randomized in factorial scheme 6 × 2 + 1, referring to the substrates [S1: DBS, S2: soil and sand 1:1 (20%) + DBS (80%), S3: soil and sand 1:1 (40%) + DBS (60%), S4: soil and sand 1:1 (60%) + DBS (40%), S5: soil and sand 1:1 (80%) + DBS (20%), S6: commercial substrate (additional treatment) and S7: soil, sand and manure 1:1:2] with and without boron. The seedling emergence, emergence rate, plant height, stem diameter, root length, root volume and dry mass of roots and shoots were recorded. The substrates S2 (without B fertilizing) and S4 (B fertilized with 0.5 mg dm^?3) produced seedlings with more than 3.0 g of shoot dry mass and at least 30 cm in height, and can be used for production of high-quality yellow passion fruit seedlings.     

Bahiagrass response and N loss from selected N fertilizer sources

The increasing cost of commercial fertilizers and environmental problems associated with improper fertilization management have prompted the need to re‐examine commercial N sources that can effectively supply N to pastures while minimizing N losses. This 3‐year study evaluated the effects of selected N sources on bahiagrass (Paspalum notatum Flügge) responses, soil properties and N losses. Treatments consisted of a factorial combination of 6 N sources [(i) ammonium nitrate (AN), (ii) ammonium sulphate (AS), (iii) urea (U), (iv) urea treated with Agrotain (U + Agrotain), (v) SuperU and (vi) ammonium sulphate nitrate] and 3 N levels (0, 60 or 120 kg ha^?1 year^?1), replicated three times. Bahiagrass dry‐matter yield (DMY), crude protein (CP) concentration, N uptake and recovery were not affected by N source, with the exception of AN that resulted in reduced DMY in 2010 compared with the other sources. Bahiagrass DMY, CP concentration, N uptake and recovery increased linearly as N levels increased. Nitrogen fertilization showed no effect on soil pH or soil N accumulation. Soil pore‐water N concentrations from treatments fertilized with N were similar to the control plots indicating no threat to the environment. At the N levels evaluated in this study, selection of N source should be based on the fertilizer cost.