Dietary tryptophan supplementation does not affect growth but increases brain serotonin level and modulates the expression of some liver genes in zebrafish (Danio rerio)

This study aimed at assessing the effects of the dietary tryptophan (Trp) supplementation on growth and feed utilization, brain serotonin content, and expression of selected liver genes (involved in the liver serotonin pathway, protein synthesis degradation, and antioxidant activity) in zebrafish. A growth trial was conducted with zebrafish juveniles fed five experimental isoproteic (40%DM) and isolipidic (8%DM) fishmeal-based diets containing graded levels of Trp: a Trp-non-supplemented diet (diet Trp0, with 0.22% Trp) and four Trp-supplemented diets containing 2–16 times higher Trp content (diets Trp2, Trp4, Trp8, and Trp16 with 0.40, 0.91, 2.02, and 3.34% Trp, respectively). Diets were tested in quadruplicate, with fish being fed twice a day, 6 days a week for 6 weeks to apparent visual satiation. At the end of the trial, growth performance and feed utilization were assessed, and fish from all experimental groups were sampled for whole-body composition analysis. In addition, fish fed low (Trp0), medium (Trp4), and high (Trp16) Trp diets were also sampled for analysis of brain serotonin content and liver gene expression. Tested tryptophan levels did not influence growth performance nor feed intake. However, values of energy and nitrogen retention as well as body energy content indicate a better feed utilization with diets containing around 0.9% and 2.0% DM Trp. Brain serotonin content increased with increasing dietary tryptophan levels. In addition, regarding liver genes, dietary treatment had a modulatory effect on the expression of Htr1aa and Htr2cl1 genes (encoding for serotonin receptors), TPH1a gene (encoding for tryptophan hydroxylase, the rate-limiting enzyme in the synthesis of serotonin from tryptophan), TOR gene (involved in protein synthesis), and Keap1 gene (involved in antioxidant responses).

     

Nutrition and production of Helianthus annuus in a function of application of leaf silicon

Silicon (Si) is important as it increases plant tolerance to abiotic stresses. However, studies on in foliar fertilization are still scarce. The objective of this research was to evaluate the effect of foliar application of silicon on nutrition and production of Helianthus annuus. The study was carried with a completely randomized design, consisting of five doses of silicon (0; 0.84; 1.68; 2.52 and 3.36?mg L^?1) applied as silicate, with four replicates. Biometric evaluations and relative chlorophyll index (RCI) measurements were performed, being also observed the dry matter production and the silicon use efficiency by plants. The RCI is not affected by the foliar application of Si, while the height and leaf area increase by 8.3 and 25.4%, respectively. Foliar Si application up to the dose of 3.36?g L^?1 promotes linear increases in the dry matter production of shoots (32%), capitulum (19%) and the whole plant of sunflower (27%).     

Seed coating with inocula of arbuscular mycorrhizal fungi and plant growth promoting rhizobacteria for nutritional enhancement of maize under different fertilisation regimes

Arbuscular mycorrhizal (AM) fungi and plant growth-promoting rhizobacteria, responsible for enhancing plant nutrition, vigour and growth, may be used to reduce dosages of chemical fertilisers. Technologies that allow an economically viable and efficient application of these beneficial microbes in large scale agriculture must be studied. Seed coating is a potential delivery system for efficiently introducing minor amounts of bioinoculants. Despite the dramatic reduction on inoculum dose per plant, inoculation of AM fungi via seed coating was as effective as conventional soil inoculation. Fertilisation and inoculation had a significant impact on maize shoots nutrient concentrations. Different fertilisation regimes did not influence mycorrhizal colonisation. Plants without fertilisation and singly inoculated with R. irregularis showed shoot nutrient concentration increments of 110, 93, 88 and 175% for nitrogen, phosphorus, potassium and zinc, respectively, comparing with non-inoculated controls. Plants singly inoculated with P. fluorescens via seed coating under full fertilisation, presented enhancements of 100, 75 and 141% for magnesium, zinc and manganese, respectively, comparing with non-inoculated controls. Seed coating is a promising tool for delivering microbial inoculants into the soil, while promoting sustainable production of maize. This technology is particularly pertinent in low input agriculture, with potential environmental profits and food quality improvements.     

Biodegradation of biodiesel and toluene under nitrate-reducing conditions and the impact on bacterial community structure

Purpose

Biodiesel is a renewable fuel that can be mixed with toluene and be accidentally released into anoxic ecosystems and impact soil microbial communities. Therefore, the aim of the present work was to examine, under nitrate-reduction conditions, the biodegradation of toluene in the presence of two different types of biodiesel (sunflower and rapeseed), and their impact on the bacterial community structure.

Materials and methods

Sediment samples were spiked individually with toluene, biodiesel, and their blends in laboratory-designed microcosms. Sunflower oil biodiesel was produced in the laboratory, while rapeseed oil biodiesel was a commercial product. Degradation of biodiesels and blends was monitored by directly measuring the substrate or indirectly by determining nitrate removal during the course of the experiment. Denitrification rates were estimated with the acetylene inhibition technique. Denaturing gradient gel electrophoresis was used to assess impacts on the bacterial community structure exposed to biodiesels, blends, and toluene.

Results and discussion

The results of this study showed that toluene and biodiesel were completely degraded within 10 days. Biodiesel significantly affected the bacterial community structure at a similar magnitude, independently of its origin. Additionally, toluene impacted the bacterial community and denitrification process to a lower extent than biodiesel and a clear decrease in the relative bacterial richness and diversity was shown in samples with biodiesel and blends. To the best of our knowledge, this is one of the first reports describing degradation of biodiesel alone and blends under nitrate-reducing conditions, and also the effects of these compounds on the denitrification process. In addition, due to the recently discovered “oxygenic denitrification” process, the acetylene inhibition technique and nitrous oxide quantification may not be the most adequate tool to estimate denitrification rates. Further detailed studies are advised to understand whether the identified bacterial community shift impacts ecosystem functions.

Conclusions

Our results help to understand the biodegradation of toluene, biodiesel, and their blends in sediments under nitrate-reducing conditions and might be important in implementing bioremediation strategies in anoxic environments.

     

Environmental factors on virulence of <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Aeromonas hydrophila are known for being opportunistic pathogens, harboring various virulence factors and triggering lesions and death in fish. The disease caused by bacteria can make fish inappropriate for human consumption, besides representing a risk to public health. The pathogenesis can be influenced by environmental variables, affecting fish productivity and mortality. The present study aimed to determine whether A. hydrophila harbor the virulence genes aerolysin, hydrolipase, elastase, lipase, cytotonic enterotoxin (ast), lateral flagellum (laf), and polar flagellum (fla) and to evaluate the influence of environmental variables on in vitro growth, in vivo virulence and expression of some of these genes. Polymerase chain reaction (PCR)-based screening for the presence of these virulence genes was performed on 35 isolates. Six isolates containing different profiles of virulence genes were tested for in vitro growth under different conditions of pH, temperature, and ammonia and for in vivo virulence under these same environmental conditions. RT-qPCR was used to quantify the expression of aerolysin, lipase, and fla genes. All the tested environmental factors influenced the growth of A. hydrophila, while pH and ammonia concentrations influenced the bacterial virulence. The expression of the fla gene increased when bacteria were grown in higher ammonia concentration. The mortality established by Aeromonas is influenced by several environmental factors pinpointing the importance of its control in fish farming to avoid higher economic loses associated to bacterial disease outbreaks.     

Apparent digestibility coefficients of European grain legumes in rainbow trout (Oncorhynchus mykiss) and Nile tilapia (Oreochromis niloticus)

Two trials were carried out to evaluate the apparent digestibility coefficients (ADCs) of dry matter (DM), crude protein (CP), lipids, starch and gross energy (GE) of six European varieties of grain legumes, namely chickpea‐type Kabuli (CHK), chickpea‐type Desi (CHD), field pea (FP), faba bean (FB), white lupin (WL) and chickling vetch (CV), in rainbow trout and Nile tilapia juveniles. The ADCs were measured using a reference diet and six experimental diets (700 g/kg of the reference diet and 300 g/kg of each raw grain legume) containing 10 g/kg chromic oxide as inert marker. Additionally, grain legumes were analysed for the organic acids profile. In rainbow trout, FP presented the highest ADCs of DM, CP and GE, whereas chickpeas and FB had the lowest DM, GE and starch ADC values. In Nile tilapia, the lowest values of nutrients (except starch) and energy ADCs were found for FP and CV. Nutrients and energy of chickpeas, WL and FB were better digested by tilapia, whereas FP was better digested by trout. Overall results reveal raw grain legumes as promising feed sources for both fish species.     

Comparative pathology of parasitic infections in free-ranging and captive pit vipers (Bothrops jararaca)

Between June 1997 and May 1998, 47 pit vipers (Bothrops jararaca) (Group A) were euthanased when they were brought to the Instituto Butantan by farmers, and examined postmortem; during the same period, 91 snakes of the same species (group B) were examined after they had died in an outdoor serpentarium. The majority of the parasites encountered were nematodes; lungworms, Rhabdias vellardi, and the intestinal hookworm Kalicephalus inermis were the most common. Some of the snakes in group A were heavily infested, but their lesions were mild, whereas in group B the parasites were generally accompanied by severe lesions. The parasites with a direct life cycle were more common than those with obligatory intermediate hosts, and the snakes were more commonly infected during the hotter and more humid seasons.     

Sero-epidemiological study of canine Leishmania spp. infection in the municipality of Alijó (Alto Douro, Portugal)

Visceral leishmaniosis caused by Leishmania infantum is a zoonotic disease in the Mediterranean basin. We report an epidemiological survey carried out in dogs from the municipality of Alijó in the endemic region of Alto Douro (north Portugal). Performance of the direct agglutination test (DAT) was assessed in 205 matching samples of blood collected on filter paper and serum. A high degree of agreement (97.6%; k = 0.83) was found between the results obtained from both types of samples. DAT was then used to test more blood on filter paper (B-FP) samples from other dogs of the same municipality. The detected sero-prevalence was 18.7% (288/1540), with values ranging from 0.0 to 81.1% in each of the 19 parishes of Alijó. Three distinct geographical zones of mean sero-prevalence could be defined: northwestern (2.5%), intermediate (11.4%) and southern (49.9%). No statistically significant difference was observed between male (19.1%) and female (17.8%) sero-prevalences (P = 0.560). Dogs of 9-11 years of age showed the highest sero-prevalence (28.4%), but all the other age-intervals (0-2, 3-5, 6-8 and 12-17 years) presented values (15.0-22.3%) not significantly different from the mean of the whole study population. Risk factors for canine Leishmania infection were age and geographical zone. Only 5.9% of the sero-positive animals had clinical signs of canine leishmaniosis and the overall prevalence of disease was 1.1%. This study validates the use of B-FP samples and confirms DAT as a simple and sensitive serological test to evaluate the level of canine Leishmania infection in areas of high sero-prevalence.