Effect of dietary sophorolipids on growth performance and gastrointestinal functionality of broiler chickens infected with Eimeria maxima.

Two experiments were conducted to evaluate the effects of dietary sophorolipids (SLs) supplementation as antibiotic alternatives on growth performance and gut health of chickens infected with Eimeria maxima. In experiment 1, 336 (zero-day-old) male broilers were used. The chickens were weighed and randomly allocated to the following 6 treatments groups with 7 chickens/cage and 8 cages/treatment: control group that received a basal diet (NC), positive control group that received a basal diet and was challenged with E. maxima (PC), PC+C18:1 lactonic diacetyled SL (SL1), PC+C18:1 deacetyled SL (SL2), PC+C18:1 monoacetyled SL (SL3), and PC+C18:1 diacetyled SL (SL4). Each SL (200 mg/kg feed) was added to the corresponding treatment group. In experiment 2, 588 (zero-day-old) male broilers were used. The chickens were randomly allocated to the following experimental groups with 10 or 11 chickens/cage and 8 cages/treatment: NC, PC, PC+ monensin at 90 mg/kg feed (MO), PC+SL1 at 200 mg/kg feed (SL1 200), PC+SL1 at 500 mg/kg feed (SL1 500), PC+SL4 at 200 mg/kg feed (SL4 200), and PC+SL4 at 500 mg/kg of feed (SL4 500). The chickens and feed were weighed at 0, 7, 14, 20, and 22 d to determine growth performance. In both experiments, all chickens except the NC group were orally infected with E. maxima (10,000 oocysts/chicken) at d 14. One chicken per cage was euthanized at d 20 to sample jejunal tissue to measure lesion scores, cytokines, and tight junction (TJ) proteins. Excreta samples were collected daily between d 20 and 22 to measure oocyst numbers. Data were analyzed using Mixed Model (PROC MIXED) in SAS. In experiment 1, SLs did not affect the growth of broiler chickens, but SL4 decreased (P < 0.05) the lesion score and oocyst number compared to PC chickens. In terms of cytokines and TJ protein gene expression, SLs increased (P < 0.05) IL-1ß, IL-6, IL-17F, IL-4, IL-13, occludin, and ZO1 levels compared to PC chickens. In experiment 2, monensin increased (P < 0.05) body weight, and decreased (P < 0.05) the lesion score and oocyst number compared to the PC group. SL4 500 increased (P < 0.05) average daily gain and feed conversion ratio but decreased (P < 0.05) lesion score and fecal oocyst number. SL4 decreased (P < 0.05) IL-6, IL-17F, TNFSF-15, IL-2, and IL-10 levels but increased (P < 0.05) occludin and ZO-1 levels. Overall, dietary SL supplementation, especially SL4, improved growth and gastrointestinal functionality of young broiler chickens, demonstrating significant potential as an antibiotic alternative.

Evaluation of the Sensitivity and Reproducibility of Targeted Proteomic Analysis Using Data-Independent Acquisition for Serum and Cerebrospinal Fluid Proteins.

There is a need for targeted analysis of biological fluids for diagnosis, prognosis, or monitoring the progression of diseases. Cerebrospinal fluid (CSF) and serum have been widely used for the development of protein analysis for neurodegenerative diseases and other diseases, respectively. Recently, data-independent acquisition (DIA) mass spectrometry (MS) has been developed to increase the throughput over data-dependent acquisition (DDA) on screening of a large number of samples and discovery of candidate targets. When it comes to target validation, the analytical performance of targeted analysis is critical. However, the inter- and intralaboratory analytical performances of the DIA-MS for targeted proteomic analysis of CSF and serum samples have not yet been investigated. In this study, we showed that the DIA-MS approach allowed us to identify and quantify 1732 CSF and 424 serum proteins, with 90% of proteins identified and quantified in at least 50% of DIA-MS runs. To evaluate the sensitivity, linearity, and dynamic range of the DIA approach, we included the stable isotope-labeled (SI) peptides into CSF and serum samples with serial dilutions. The lower limit of quantification (LLOQ) of peptides was 0.1-0.5 fmol, and the dynamic range was over 3.53 orders of magnitude, with excellent linearity (r2 < 0.978) in CSF and serum samples. Finally, the reproducibility of the DIA-MS approach was evaluated using entire proteins identified in CSF and serum samples. The intralaboratory three replicate results showed reliable reproducibility with 12.5 and 17.3% of the median coefficient of variation (CV) in both CSF and serum matrices, whereas the median CVs of interlaboratory three replicates were 23.8 and 32.0% in CSF and serum samples, respectively. The comparison of the quantitative result between replicates showed close similarity at intra- and interlaboratories with a median Pearson correlation value of >0.98 in CSF and serum, respectively. In conclusion, we demonstrate the capability of the DIA approach as a targeted proteomic analysis for candidate proteins from CSF and serum samples.

C9orf72 regulates energy homeostasis by stabilizing mitochondrial complex I assembly.

The haploinsufficiency of C9orf72 is implicated in the most common forms of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), but the full spectrum of C9orf72 functions remains to be established. Here, we report that C9orf72 is a mitochondrial inner-membrane-associated protein regulating cellular energy homeostasis via its critical role in the control of oxidative phosphorylation (OXPHOS). The translocation of C9orf72 from the cytosol to the inter-membrane space is mediated by the redox-sensitive AIFM1/CHCHD4 pathway. In mitochondria, C9orf72 specifically stabilizes translocase of inner mitochondrial membrane domain containing 1 (TIMMDC1), a crucial factor for the assembly of OXPHOS complex I. C9orf72 directly recruits the prohibitin complex to inhibit the m-AAA protease-dependent degradation of TIMMDC1. The mitochondrial complex I function is impaired in C9orf72-linked ALS/FTD patient-derived neurons. These results reveal a previously unknown function of C9orf72 in mitochondria and suggest that defective energy metabolism may underlie the pathogenesis of relevant diseases.

Growth-Promoting and Antioxidant Effects of Magnolia Bark Extract in Chickens Uninfected or Co-Infected with Clostridium perfringens and Eimeria maxima as an Experimental Model of Necrotic Enteritis.

BACKGROUND: Magnolia tree bark has been widely used in traditional Asian medicine. However, to our knowledge, no studies have been reported investigating the effects of dietary supplementation with magnolia bark extract in chickens. OBJECTIVE: We tested the hypothesis that dietary supplementation of chickens with a Magnolia officinalis bark extract would increase growth performance in uninfected and Eimeria maxima/Clostridium perfringens co-infected chickens. METHODS: A total of 168 chickens were fed from hatch either a standard diet or a diet supplemented with 0.33 mg or 0.56 mg M. officinalis bark extract/kg (M/H low or M/H high, respectively) from days 1 to 35. At day 14, half of the chickens were orally infected with E. maxima, followed by C. perfringens infection at day 18 to induce experimental avian necrotic enteritis. Daily feed intake, feed conversion ratio, body weight gain, and final body weight were measured as indicators of growth performance. Serum α1-acid glycoprotein (AGP) concentrations were measured as an indicator of systemic inflammation, and intestinal lesion scores were determined as a marker of disease progression. Transcript levels for catalase, heme oxygenase 1, and superoxide dismutase in the intestine, liver, spleen, and skeletal muscle were measured as indicators of antioxidant status. RESULTS: Growth performance increased between days 1 and 35 in uninfected and E. maxima/C. perfringens co-infected chickens fed M/H-low or M/H-high diets compared with unsupplemented controls. Gut lesion scores were decreased, whereas AGP concentrations were unchanged, in co-infected chickens fed magnolia-supplemented diets compared with unsupplemented controls. In general, transcripts for antioxidant enzymes increased in chickens fed magnolia-supplemented diets compared with unsupplemented controls, and significant interactions between dietary supplementation and co-infection were observed for all antioxidant enzyme transcript levels. CONCLUSION: Magnolia bark extract might be useful for future development of dietary strategies to improve poultry health, disease resistance, and productivity without the use of antibiotic growth promoters.

Effects of dietary Allium hookeri root on growth performance and antioxidant activity in young broiler chickens.

The objective of this experiment was to evaluate the antioxidant potential of Allium hookeri (AH) root in two forms, powdered AH root and fermented powdered AH root, to demonstrate its value as an antibiotic alternative feed additive for broiler chickens. A total of 125 male Ross-708 broiler chickens were randomly assigned to five groups (n = 25 birds/group) and fed standard diets supplemented with root or fermented root of AH at two different levels (1% or 5%). Control birds were provided with non-supplemented basal diets. Body weights was measured at days 14 and 21 of age. To monitor antioxidant activities, heme oxygenase (HMOX), aflatoxin B1 aldehyde reductase (AFAR), superoxide dismutase 1 (SOD1), and catalase (CAT) enzyme levels were quantified by real-time PCR in the jejunums 21-day-old birds. Also, serum levels of SOD, CAT, and malondialdehyde (MDA) were measured using commercial kits. The results showed greater body weight gains at day 14 in chickens fed diets supplemented with 1% AH root, as compared to the control group (p < 0.05). Up-regulated transcript levels of AFAR, HMOX1, and CAT were observed in the jejunum of chickens fed diets supplemented with AH. The serum levels SOD and CAT were significantly increased (p < 0.05) in groups treated with AH, whereas MDA levels were decreased in groups fed diets supplemented with AH, as compared to the control group. These results indicated that an optimum level of dietary AH supplementation to young broiler chickens influences growth and improves antioxidant activities.

Dietary Allium hookeri reduces inflammatory response and increases expression of intestinal tight junction proteins in LPS-induced young broiler chicken.

We undertook a study to assess the effects of Allium hookeri (AH) root and fermented root on inflammation and intestinal integrity of lipopolysaccharide (LPS)-challenged broiler chickens. Birds were assigned to six groups (n = 25 birds/treatment) and fed with basal diets (CON) or basal diets supplemented with AH root or fermented root at two concentrations (1 or 5%). At 7 d of age, five groups (n= 125) in each dietary treatment were injected with LPS (1 mg/kg body weight), and the remaining 25 birds were injected with sterile phosphate-buffered saline (PBS) as a negative control. LPS challenge significantly reduced average body weight gain at 24 h post-injection compared with PBS control. Fermented root supplementation increased average body weight gain by 1% compared with the LPS-challenged control. Serum α- 1-AGP levels, interleukin (IL)-1ß, IL-8, tumor necrosis factor superfamily member 15 (TNFSF15), and LPS-induced tumor necrosis factor-α factor (LITAF) transcript levels were significantly higher in the small intestine in LPS-injected chickens. However α-1-AGP levels were reduced by AH root or fermented root (1 and 5%) supplementation and IL-1ß, IL-8, and LITAF were also down-regulated by root and fermented root (1 and 5%) supplementation. The reduced expression of tight junction proteins (junctional adhesion molecule 2 (JAM2) and occludin) and intestinal mucin 2 (MUC2) by LPS challenge was reversed by root or fermented root (1 and 5%) supplementation. These findings demonstrate that dietary AH root and fermented root influence antiinflammatory activity and tight junction protein expression in LPS-induced chickens.