Immune responses and gut morphology of Senegalese sole (Solea senegalensis,Kaup 1858) fed monospecies and multispecies probiotics

The current study aimed to determine the effects of dietary probiotic supplementation on growth, gut morphology and non‐specific immune parameters in Senegalese sole (Solea senegalensis) juveniles during a 1‐month trial. Fish were fed for 1‐month two diets with 1.0 or 4.6 × 10⁶ CFU kg^?1) of probiotic A (Bacillus sp., Pediococcus sp., Enterococcus sp. and Lactobacillus sp.) and two diets with 3.5 or 8.6 × 10⁵ CFU kg^?1 of probiotic B (Pediococcus acidilactici) and tested against an unsupplemented diet (control). Growth performance, as well as respiratory burst activity, nitric oxide (NO), alternative complement pathway (ACH50), lysozyme and peroxidase activities, was not affected by the dietary treatments. Probiotic supplementation tended to increased growth homogeneity between tanks having diet A₁ the best possible alternative to decrease costs associated to size grading. Villous length and number of goblet cells of the anterior intestine did not vary among treatments. Muscle duodenal layer was significantly thicker in fish fed probiotic A compared to probiotic B, when included at the lowest level (A₂ versus B₂). The current study indicate that the use of the multispecies probiotic at 1.0 × 10⁶ CFU kg^?1 might enhance protection against pathogen outbreak and increase nutrient absorption, whereas at the highest concentration could reduced size dispersion among tanks.     

Protein content and amino acid composition of eggs and endotrophic larvae from wild and captive fed sea urchin Paracentrotus lividus (Echinodermata: Echinoidea)

Knowledge on the biochemistry of proteins and amino acids (AA) of eggs and larvae of echinoids is scarce and the possibility to modify their profiles by diet manipulations is unknown. The protein content of eggs, prisms and pre‐plutei and the amino acid composition of eggs of Paracentrotus lividus from captive broodstock fed prepared diets were analysed and compared with the ones obtained from wild broodstock. Diets differed on protein source (fish or soy meals) and on protein content (10–40% dry weight – DW). Total and soluble protein content of both eggs and larvae was higher than 400 g kg^?1 DW and 200 g kg^?1 DW respectively. Glycine was the most abundant free AA. Very few differences were found among P. lividus eggs and larvae biochemical parameters, suggesting that their AA composition can hardly be changed by broodstock diet manipulations.     

Feed can be a source of Campylobacter jejuni infection in broilers

1. The aim was to determine the importance of a contaminated diet as a possible cause of Campylobacter jejuni infection in broilers.

2. This study evaluated the viability of C. jejuni in both starter and finisher diets and the interference from other mesophilic bacteria in this viability.

3. Starter and finisher samples of broiler diet were deliberately contaminated with 3 or 5 log CFU·g^?1 of C. jejuni (NCTC 11351) and then maintained at two different storage temperatures (25°C or 37°C) for 3 or 5 d.

4. C. jejuni survived during this period and, when inoculated at 10³ CFU·g^?1, multiplied with greater proliferation at a storage temperature of 37°C. There was no relationship between the amount of mesophilic bacteria and C. jejuni viability.

5. This study highlights the importance of the diet in the epidemiology of C. jejuni in broilers.     

Neospora caninum DNA detection by TaqMan real-time PCR assay in experimentally infected pregnant heifers

Neosporosis has been considered the main cause of abortion between the first and the second trimester of pregnancy in cattle. Therefore, the objective of this study was to identify the presence of Neospora caninum DNA obtained from experimental models based on the evaluation of different areas of the fetal nervous system and organs from heifers previously inoculated with NC-1 after or before insemination. This study was performed with Hereford × Nelore (n = 29) heifers and all animals were considered free of diseases at the beginning of the experiment. All animals were bred by fixed-time artificial insemination (TAI) and allocated as follows: (a) seronegative heifers subjected to TAI (TAI, n = 9), (b) heifers infected with N. caninun 60 days prior to TAI (NC-1 + TAI, n = 9), and (c) heifers submitted to TAI and infected with N. caninum 60 days later (TAI + NC-1, n = 11). The pregnancy was confirmed by transrectal ultrasonography 35 days after TAI and evaluated every 30 days until the end of gestation. Fetuses were collected surgically at 170 days of gestation, and immediately necropsied to remove tissues aseptically. Samples of the central nervous system (CNS), heart, kidney, lung, liver, skeletal muscle and caruncle were collected for DNA extraction. Days of gestation at abortion and interval from abortion to first insemination were examined by Student's t-test. At 35 days of gestation the pregnancy rates in the group NC-1 + TAI (4/9, 44.4%) was lower than in the control group (8/9, 88.8%, P < 0.05). At 60 days, the pregnancy rates in the NC-1 + TAI group (0/4, 0%) was lower compared to TAI + NC-1 (5/7, 71.4%) and control (6/8, 75.0%) groups (P < 0.05). Animals from the group NC-1 + TAI were re-inseminated 60 days after the first TAI. After pregnancy losses throughout the study, 5 animals (TAI), 3 animals (NC-1 + TAI) and 5 animals (TAI + NC-1) maintained pregnancy until 170 days of gestation. TaqMan RT-PCR demonstrated the presence of N. caninum DNA in the medulla and right posterior cortex in 3 out of 5 fetuses from the TAI + NC-1 group. We concluded that heifers infected after TAI had a higher incidence of the parasite at the fetus CNS. Identification of N. caninum by TaqMan RT-PCR would assist in the investigation of infection and in the evaluation of vaccines or therapeutic drugs to control neosporosis in cattle.     

Incidence of subclinical endometritis and its effects on reproductive performance of crossbred dairy cows

In dairy cattle, uterine infections are not life threatening and often unavoidable; however, they reduce fertility and increase the production costs of properties. The aim of this study was to evaluate the incidence of subclinical endometritis from 32 to 70 days in milk (DIM) and its effects on the reproductive performance of crossbred dairy cows. Lactating cows (Holstein/Gir; n?=?172), with no history of retained placenta, without clinical signs of uterine infection were used. The body condition score (BCS) was evaluated on a scale from 1 to 5. Ultrasound examination was performed to evaluate uterine lining and ovarian activity, while vaginal mucus was analyzed by gloved hand. The diagnosis of subclinical endometritis was performed by endometrial cytobrush technique. The samples were collected, stained, and examined microscopically; positive cases for subclinical endometritis were considered with the presence of ≥5 % of neutrophils. Later, the cows were submitted to conventional artificial insemination or timed artificial insemination. The incidence of subclinical endometritis in the herd was 26 %, and this was not affected by the season of calving, presence of corpus luteum, DIM, and parity. Cows with a BCS ≤2.50 had a higher incidence of subclinical endometritis. The conception rate to first insemination and pregnancy rate at 150 days postpartum were not influenced by the presence of subclinical endometritis in crossbred dairy cows.     

Metal contamination effects on sunflower (Helianthus annuus L.) growth and protein expression in leaves during development

Metal-ion contamination (Cd, Cu, Pb, and Zn) on sunflower (Helianthus annuus L.) growth and total leaf protein expression were studied in the present work. The height, mass production, and metal distribution (Ca, K, Fe, Mg, Na, and P) in all plant fractions (roots, stems, and leaves) were evaluated. Sunflowers plants contaminated with four metal ions decreases height and mass by 35% and 40%, respectively, compared to control. Significant differences of total protein composition were noted after SDS-PAGE separation. Sunflower proteomics were more affected when 500 mg L(-1) of metal ion was added as contaminant of both zinc and mixed ions solution. In these cases, proteins having a molar mass of 14.5, 34.5, and 54.0 kDa were present at a lower level and alterations in enzymatic activities (SOD and GR) were found. Sunflowers plants contaminated with zinc and the mixed ions solution showed some degree of oxidative stress.     

Effects of site preparation and slash management on growth and understory vegetation of Eucalyptus globulus plantations along a rotation time span in Portugal

The removal or maintenance of harvest residues in eucalypt plantations may influence site quality and productivity. Removal of slash from the site may facilitate further management operations and provide a valuable energy resource, but effects on site productivity and sustainability for a rotation time span were not yet assessed under Mediterranean conditions. Therefore, a study was carried out to assess the effects of slash (harvest residues plus forest floor litter) management and soil preparation options on stand productivity and understory vegetation dynamics, hypothesizing that those options influence tree growth, forest floor dynamics and understory biomass and diversity. An experiment was installed in West Central Portugal, consisting on: removal of slash without soil preparation (R); broadcast of harvest residues on the soil surface without soil disturbance (S); as in S, but concentrating the woody debris between tree rows (W); incorporation of slash into the soil by harrowing (I); removal of slash followed by harrowing (RH); and as in I followed by ripping (IS). The experiment was monitored for a rotation time span (140 months). Maintenance of slash followed by deep soil disturbance led to the highest wood production, but differences between treatments were not significant (p > 0.05). Forest floor load and understory biomass were also similar between treatments. Ground vegetation played an important role on nutrient cycling in early rotation stages, such effect being irrespective on slash management options. Incorporation of slash into the soil followed by ripping is probably the best option to match production and environmental sustainability of eucalypt plantations in Mediterranean conditions.     

Angiotensin-converting enzyme inhibition by Brazilian plants

The potential antihypertensive activity of Brazilian plants was evaluated in vitro by its ability to inhibit the angiotensin-converting enzyme (ACE). Forty-four plants belonging to 30 families were investigated. Plants were selected based on their popular use as antihypertensive and/or diuretics. The following plants presented significant ACE inhibition rates: Calophyllum brasiliense, Combretum fruticosum, Leea rubra, Phoenix roebelinii and Terminalia catappa.     

Tick saliva induces regulatory dendritic cells: MAP-kinases and Toll-like receptor-2 expression as potential targets

Ticks (Acari: Ixodidae) are bloodsucking ectoparasitic arthropods of human and veterinary medical importance. Tick saliva has been shown to contain a wide range of bioactive molecules with vasodilatory, antihemostatic, and immunomodulatory activities. We have previously demonstrated that saliva from Rhipicephalus sanguineus ticks inhibits the maturation of dendritic cells (DCs) stimulated with LPS. Here we examined the mechanism of this immune subversion, evaluating the effect of tick saliva on Toll-like receptor (TLR)-4 signalling pathway in bone marrow-derived DCs. We demonstrated that R. sanguineus tick saliva impairs maturation of DCs stimulated with LPS, a TLR-4 ligand, leading to increased production of interleukin (IL)-10 and reduced synthesis of IL-12p70 and TNF-α. The immunomodulatory effect of the tick saliva on the production of pro-inflammatory cytokines by DCs stimulated with LPS was associated with the observation that tick saliva inhibits the activation of the ERK 1/2 and p38 MAP kinases. These effects were independent of the expression of TLR-4 on the surface of DCs. Additionally, saliva-treated DCs also presented a similar pattern of cytokine modulation in response to other TLR ligands. Since the recent literature reports that several parasites evade immune responses through TLR-2-mediated production of IL-10, we evaluated the effect of tick saliva on the percentage of TLR-2⁺ DCs stimulated with the TLR-2 ligand lipoteicoic acid (LTA). The data showed that the population of DCs expressing TLR-2 was significantly increased in DCs treated with LTA plus saliva. In addition, tick saliva alone increased the expression of TLR-2 in a dose- and time-dependent manner. Our data suggest that tick saliva induces regulatory DCs, which secrete IL-10 and low levels of IL-12 and TNF-α when stimulated by TLR ligands. Such regulatory DCs are associated with expression of TLR-2 and inhibition of ERK and p38, which promotes the production of IL-10 and thus down-modulates the host's immune response, possibly favouring susceptibility to tick infestations.     

Dietary taurine supplementation in plant protein based diets do not affect growth and reproductive performance of zebrafish

Taurine (Tau) has been regarded as a conditional essential nutrient for some fish species. Although its role has been extensively studied in higher vertebrates, limited results are reported with fish especially its role on reproductive performance and the ontogenic changes on Tau levels throughout the life cycle. Therefore, we designed a feeding trial using zebrafish as a model species to test whether Tau supplementation to plant protein diets would have a positive effect on growth and reproductive performance. Zebrafish were fed plant protein diets containing graded levels of Tau (0.2, 4.6, 5.9 and 13.7 g/kg diet) from 10 days post fertilization (dpf) to sexual maturity. An additional commercial diet was used as a positive control for performance. The trial followed a completely randomized design with five treatments (diets) and three replications. After 60 days of feeding, growth, Tau concentration in the body, redox status, lipid body composition, reproductive and offspring performances were analysed. Tau supplementation did not affect growth and/or reproductive performance; however, zebrafish seems to differently modulate Tau concentration according to the growth stage. Tau seemed to induce a hypolipidemic effect in zebrafish by reducing lipid accumulation in their bodies (p < .05). A trend to a more pro‐oxidant effect of Tau supplementation was observed by the decreased reduced glutathione levels. In sum, Tau does not affect growth and reproductive performance of zebrafish but it is important for normal lipid utilization and redox status.