Effects of crude protein and lactose levels in diets on growth performance,intestinal morphology,and expression of genes related to intestinal integrity and immune system in weaned piglets

To evaluate the effects of crude protein (CP) and lactose (LAC) for weaned piglets on performance, intestinal morphology, and expression of genes related to intestinal integrity and immune system, 144 piglets with initial weight 7.17 ± 0.97 kg were allotted in a randomized design, in a 2 × 3 factorial arrangement (20.0% and 24.0% CP and 8.0%, 12.0%, and 16.0% LAC) with eight replicates. Piglets fed 20.0% CP had greater weight gain and feed intake. Including 12.0% LAC in the 20.0% CP diet provided higher villous height in the duodenum than 8.0% LAC, and 12.0% or 16.0% LAC in the 24.0% CP diet resulted in higher villous height in the jejunum and ileum, and higher villi/crypt ratio in the ileum than 8.0% LAC. No effects of CP and LAC on interleukin‐1β and tumor necrosis factor‐α mRNA were observed. The 16.0% LAC diet provided higher gene expression of transforming‐β1 growth factor. Feeding 20.0% CP resulted in better performance than 24.0% CP. The 12.0% LAC diet promoted greater genetic expression of occludin and zonula occludens. Including 12.0% LAC in the diet may improve intestinal epithelial morphology and integrity, and these improvements are more evident when piglets are fed diets with 24.0% CP.     

A metabarcoding tool to detect predation of the honeybee Apis mellifera and other wild insects by the invasive Vespa velutina

The invasive Vespa velutina has been widely referred as an effective predator of honeybees. Despite the potential risk to pollination services provision and honey production, there is no accurate quantification and assessment of its real consequences for honeybees. To date, the identification of the honeybee and other insects in the diet of V. velutina has been investigated by direct observation of adult foraging or examination of food pellets. To overcome these limitations, in this study we used a DNA metabarcoding approach to evaluate the usefulness of different types of sample (jaws and stomachs collected from workers and larval faecal pellets taken from the hornet comb) to investigate the predation of V. velutina upon honeybees, and potentially on other insects. Honeybee DNA was identified in all types of samples, but larval faecal pellets retrieved the higher number of reads of honeybee DNA and the largest diversity at all taxonomic levels. Over all samples we could identify 4 orders, 9 families, 6 genera and 1 species of prey. We estimate that collecting 6 workers is sufficient to identify honeybee predation by a colony using worker’s jaws. Stomachs were the least useful sample type to detect honeybee DNA. The presence of honeybee DNA in all analysed colonies irrespective of collection site, and the variety of insect orders detected in the diet support current concerns over the acknowledged negative impact of V. velutina on managed honeybees and its potential threat to pollination services provision.

     

Effects of putrescine injection in broiler breeder eggs

The aim of this study was to evaluate the effects of increasing doses of putrescine injected in ovo on hatchability, intestinal morphology and pre-starter performance of broilers. For this purpose, 720 eggs from broiler breeders were separated into a negative control (no injection) and injection treatments with increasing doses of putrescine (0.05; 0.1; 0.15 and 0.2%), totalling five treatments of 144 eggs each. Eggs were distributed in a completely randomized design inside the setter and the injection of solutions occurred at 17 days of incubation. After hatch, 330 birds were housed in mixed lots following the original treatments, totalling 5 treatments of 6 replicates with 11 birds each. Six birds per treatment were weighed and euthanized by cervical dislocation to collect the liver, intestine and breast 24 hr after injection, at hatch and 24 hr after hatch. At 2 days of age, intestines were collected from 4 animals per treatment to analyse histomorphology. The effects of putrescine levels were evaluated by polynomial regression models, ANOVA and Tukey test at 5% probability. The hatchability decreased linearly in response to increased doses of putrescine. The percentage of residual yolk was lower in animals that received putrescine compared to the control. After injection, the percentage of breast increased linearly, and the percentage of intestine had a quadratic response to increased doses of putrescine. However, 24 hr after hatch, the percentage of intestine linearly decreased, and the percentage of liver linearly increased in response to increased doses of putrescine. Villus height increased quadratically, crypt depth decreased linearly, and goblet cells increased linearly in response to the putrescine dose. FI and BWG were not affected in the pre-starter phase; however, FCR increased in response to increased levels of putrescine. Due to putrescine effects on embryos, it is recommended that the doses injected in ovo not exceed 0.1%.     

An iron-acquisition-deficient mutant of Corynebacterium pseudotuberculosis efficiently protects mice against challenge

Caseous lymphadenitis (CLA) is a chronic disease that affects sheep and goats worldwide, and its etiological agent is Corynebacterium pseudotuberculosis. Despite the economic losses caused by CLA, there is little information about the molecular mechanisms of bacterial pathogenesis, and current immune prophylaxis against infection has been unable to reduce the incidence of CLA in goats. Recently, 21 different mutant strains of C. pseudotuberculosis were identified by random mutagenesis. In this study, these previously generated mutants were used in mice vaccination trials to develop new immunogens against CLA. Based on this analysis, {"type":"entrez-nucleotide","attrs":{"text":"CZ171053","term_id":"58296493","term_text":"CZ171053"}}CZ171053, an iron-acquisition-deficient mutant strain, was selected. After challenge with a virulent strain, 80% of the animals that were immunized with the {"type":"entrez-nucleotide","attrs":{"text":"CZ171053","term_id":"58296493","term_text":"CZ171053"}}CZ171053 strain survived. Furthermore, this vaccination elicited both humoral and cellular responses. Intracellular survival of the bacterium was determined using murine J774 cells; in this assay, the {"type":"entrez-nucleotide","attrs":{"text":"CZ171053","term_id":"58296493","term_text":"CZ171053"}}CZ171053 had reduced intracellular viability. Because iron acquisition in intracellular bacteria is considered one of their most important virulence factors during infection, these results demonstrate the immunogenic potential of this mutant against CLA.     

New sources of resistance to Meloidogyne incognita race 3 in wild cotton accessions and histological characterization of the defence mechanisms

Accessions of Gossypium spp., some of them never previously tested, were evaluated for resistance to a local isolate of Meloidogyne incognita race 3 in greenhouse experiments. Nematode infection was characterized based on the galling and egg mass indexes and the reproduction factors (RF). Root‐knot nematode reproduction among the newly tested accessions varied from highly susceptible (AS0188, AS0189) to moderately resistant (MT123 no. 3), and some accessions showed highly reduced nematode reproduction (CIR1343, CIR1348, Fai Mui). Histological observations of two resistant accessions (G. barbadense CIR1348 and G. hirsutum TX‐25, respectively) showed that resistance occurs through a two‐stage mechanism in the first accession and through a single‐stage mechanism in the second. Parasitism is blocked early after second‐stage juvenile (J2) penetration or during its initial tissue migration (CIR1348) and the development of later‐stage juveniles into female adults is suppressed at a later stage (TX‐25 and CIR1348). Fluorescence and bright light microscopy showed that root cells surrounding nematodes exhibit a hypersensitivity‐like reaction, with the accumulation of presumably phenolic compounds and the presence of necrotic cells that limit the development of nematodes and the formation of giant cells. Underdeveloped giant cells with degenerated cytoplasmic content were found in small numbers in CIR1348 and in large numbers in TX‐25, along with deformed nematodes. The full characterization of the defence mechanisms of novel sources of resistance to the root‐knot nematode in cotton constitutes a first step towards their use in crop improvement.     

Mucosal and systemic T cell response in mice intragastrically infected with Neospora caninum tachyzoites

The murine model has been widely used to study the host immune response to Neospora caninum. However, in most studies, the intraperitoneal route was preferentially used to establish infection. Here, C57BL/6 mice were infected with N. caninum tachyzoites by the intragastric route, as it more closely resembles the natural route of infection through the gastrointestinal tract. The elicited T-cell mediated immune response was evaluated in the intestinal epithelium and mesenteric lymph nodes (MLN). Early upon the parasitic challenge, IL-12 production by conventional and plasmacytoid dendritic cells was increased in MLN. Accordingly, increased proportions and numbers of TCRαβ⁺CD8⁺IFN-γ⁺ lymphocytes were detected, not only in the intestinal epithelium and MLN, but also in the spleen of the infected mice. In this organ, IFN-γ-producing TCRαβ⁺CD4⁺ T cells were also found to increase in the infected mice, however later than CD8⁺ T cells. Interestingly, splenic and MLN CD4⁺CD25⁺ T cells sorted from infected mice presented a suppressive activity on in vitro T cell proliferation and cytokine production above that of control counterparts. These results altogether indicate that, by producing IFN-γ, TCRαβ⁺CD8⁺ cells contribute for local and systemic host protection in the earliest days upon infection established through the gastrointestinal tract. Nevertheless, they also provide substantial evidence for a parasite-driven reinforcement of T regulatory cell function which may contribute for parasite persistence in the host and might represent an additional barrier to overcome towards effective vaccination.     

Influence of mixtures of acenaphthylene and benzo[a]anthracene on their degradation by Pleurotus ostreatus in sandy soil

Purpose

Polycyclic aromatic hydrocarbons (PAHs) are a class of organic compounds commonly found as soil contaminants. Fungal degradation is considered as an environmentally friendly and cost-effective approach to remove PAHs from soil. Acenaphthylene (Ace) and Benzo[a]anthracene (BaA) are two PAHs that can coexist in soils; however, the influence of the presence of each other on their biodegradation has not been studied. The biodegradation of Ace and BaA, alone and in mixtures, by the white rot fungus Pleurotus ostreatus was studied in a sandy soil.

Materials and methods

Experimental microcosms containing soil spiked with different concentrations of Ace and BaA were inoculated with P. ostreatus. Initial (t ₀) and final (after 15 days of incubation) soil concentrations of Ace and BaA were determined after extraction of the PAHs.

Results and discussion

P. ostreatus was able to degrade 57.7% of the Ace in soil spiked at 30 mg kg^?1 dry soil and 65.8% of Ace in soil spiked at 60 mg kg^?1 dry soil. The degradation efficiency of BaA by P. ostreatus was 86.7 and 77.4% in soil spiked with Ace at 30 and 60 mg kg^?1 dry soil, respectively. After 15 days of incubation, there were no significant differences in Ace concentration between soil spiked with Ace and soil spiked with Ace + BaA, irrespective of the initial soil concentration of both PAHs. There were also no differences in BaA concentration between soil spiked with BaA and soil spiked with BaA + Ace.

Conclusions

The results indicate that the fungal degradation of Ace and BaA was not influenced by the presence of each other’s PAH in sandy soil. Bioremediation of soils contaminated with Ace and BaA using P. ostreatus is a promising approach to eliminate these PAHs from the environment.     

Stepwise quenching of exciton fluorescence in carbon nanotubes by single-molecule reactions

Single-molecule chemical reactions with individual single-walled carbon nanotubes were observed through near-infrared photoluminescence microscopy. The emission intensity within distinct submicrometer segments of single nanotubes changed in discrete steps after exposure to acid, base, or diazonium reactants. The steps were uncorrelated in space and time and reflected the quenching of mobile excitons at localized sites of reversible or irreversible chemical attack. Analysis of step amplitudes revealed an exciton diffusional range of about 90 nanometers, independent of nanotube structure. Each exciton visited about 10,000 atomic sites during its lifetime, providing highly efficient sensing of local chemical and physical perturbations.