Non-challenged Group Research Articles

Mitigating impact of Glycyrrhiza glabra on virulent Newcastle disease virus challenge in chickens: clinical studies, histopathological alterations and molecular docking.

Newcastle disease (ND) is widely regarded as one of the most virulent and destructive viral infections that create chaos in the poultry industry and cause widespread epidemics and consequentially debilitating economic losses on a global scale in terms of chicken products. The current experiment evaluates the protective effect of Glycyrrhiza glabra ( G. glabra) against the Newcastle disease virus (NDV) in chickens. Ninety (90) 1-day-old SPF chicks were treated according to ethical approval (BUFVTM 05-02-22) as follows (1) non-treated non-challenged control group; (2) NDV group: Challenged with genotype VII ND virus; and (3) LE/NDV group: Challenged with the virus and intermittently treated with powdered extract of G. glabra roots (LE) in drinking water (0.5g/L) before and after viral challenge. The water medication of NDV-challenged chicks has resulted in a significant decrease in the severity of clinical symptoms, morbidity, and mortality rates, as well as the quantity of virus shed, compared with the NDV group. Treatment with LE has led to a significant reduction in serum ALT and AST activities, blood glucose level, urea, and creatinine, and significant restoration of serum proteins. In addition, the treatment has resulted in a decrease in MDA and NO levels, as well as an increase in T-SOD and catalase activities compared with untreated challenged chicks. LE decreased IFN-γ and TLR-3 gene expression in comparison with the NDV group. The treated challenged birds had fewer macroscopically detectable lesions in their respiratory, digestive, and lymphoid organs than the untreated challenged birds. Microscopically, the LE/NDV group exhibited mild to moderate pathological changes in the respiratory and digestive systems as well as lymphoid tissues, in contrast to the NDV group, which exhibited severe pathological changes. Furthermore, molecular docking assessment proved the efficacy of G. glabra against viral proliferation and invasion. We concluded that Glycyrrhiza glabra powdered extract at a dose of 0.5g/L drinking water can effectively mitigate the debilitating effects of Newcastle disease in chickens.

Immunization with an mRNA DTP vaccine protects against pertussis in rats.

Bordetella pertussis is a Gram-negative bacterium that is the causative agent of the respiratory disease known as pertussis. Since the switch to the acellular vaccines of DTaP and Tap, pertussis cases in the US have risen and cyclically fallen. We have observed that mRNA pertussis vaccines are immunogenic and protective in mice. Here, we further evaluated the pertussis toxoid mRNA antigen and refined the formulation based on optimal pertussis toxin neutralization in vivo. We next evaluated the mRNA pertussis vaccine in Sprague-Dawley rats using an aerosol B. pertussis challenge model paired with whole-body plethysmography to monitor coughing and respiratory function. Female Sprague-Dawley rats were primed and boosted with either commercially available vaccines (DTaP or wP-DTP), an mRNA-DTP vaccine, or mock-vaccinated. The mRNA-DTP vaccine was immunogenic in rats and induced antigen-specific IgG antibodies comparable to DTaP. Rats were then aerosol challenged with a streptomycin-resistant emerging clinical isolate D420Sm1. Bacterial burden was assessed at days 1 and 9 post-challenge, and the mRNA vaccine reduced burden equal to both DTaP and wP-DTP. Whole-body plethysmography revealed that mRNA-DTP vaccinated rats were well protected against coughing which was comparable to the non-challenged group. These data suggest that an mRNA-DTP vaccine is immunogenic in rats and provides protection against aerosolized B. pertussis challenge in Sprague-Dawley rats.

Novel Synergistic Probiotic Intervention: Transcriptomic and Metabolomic Analysis Reveals Ameliorative Effects on Immunity, Gut Barrier, and Metabolism of Mice during Salmonella typhimurium Infection.

Salmonella typhimurium (S. typhimurium), a prevalent cause of foodborne infection, induces significant changes in the host transcriptome and metabolome. The lack of therapeutics with minimal or no side effects prompts the scientific community to explore alternative therapies. This study investigates the therapeutic potential of a probiotic mixture comprising Lactobacillus acidophilus (L. acidophilus 1.3251) and Lactobacillus plantarum (L. plantarum 9513) against S. typhimurium, utilizing transcriptome and metabolomic analyses, a novel approach that has not been previously documented. Twenty-four SPF-BALB/c mice were divided into four groups: control negative group (CNG); positive control group (CPG); probiotic-supplemented non-challenged group (LAPG); and probiotic-supplemented Salmonella-challenged group (LAPST). An RNA-sequencing analysis of small intestinal (ileum) tissue revealed 2907 upregulated and 394 downregulated DEGs in the LAPST vs. CPG group. A functional analysis of DEGs highlighted their significantly altered gene ontology (GO) terms related to metabolism, gut integrity, cellular development, and immunity (p ≤ 0.05). The KEGG analysis showed that differentially expressed genes (DEGs) in the LAPST group were primarily involved in pathways related to gut integrity, immunity, and metabolism, such as MAPK, PI3K-Akt, AMPK, the tryptophan metabolism, the glycine, serine, and threonine metabolism, ECM-receptor interaction, and others. Additionally, the fecal metabolic analysis identified 1215 upregulated and 305 downregulated metabolites in the LAPST vs. CPG group, implying their involvement in KEGG pathways including bile secretion, propanoate metabolism, arginine and proline metabolism, amino acid biosynthesis, and protein digestion and absorption, which are vital for maintaining barrier integrity, immunity, and metabolism. In conclusion, these findings suggest that the administration of a probiotic mixture improves immunity, maintains gut homeostasis and barrier integrity, and enhances metabolism in Salmonella infection.

Different methionine to cysteine supplementation ratios altered bone quality of broilers with or without Eimeria challenge assessed by dual energy X-ray absorptiometry and microtomography

Despite the acknowledged significance of nutrition in bone development, effects of methionine (Met) and cysteine (Cys) on bone quality remain under-researched, particularly during Eimeria challenge. We investigated the effects of different supplemental Met to Cys ratios (MCR) on bone quality of broilers under Eimeria challenge. A total of 720 fourteen-day old Cobb500 broilers were allocated into a 5 × 2 factorial arrangement. Five diets with Met and Cys supplemented at MCR of 100:0, 75:25, 50:50, 25:75, and 0:100 were fed to the birds with or without Eimeria challenge. Body composition was measured by dual energy x-ray absorptiometry, and the femur bone characteristics were assessed by microtomography. Data were analyzed by two-way ANOVA and orthogonal polynomial contrast. The results reaffirmed the detrimental effects of Eimeria challenge on bone quality. On 9 d post inoculation (DPI), significant interaction effects were found for whole body bone mineral content (BMC), lean tissue weight, and body weight (P < 0.05); in the nonchallenged group (NCG), these parameters linearly decreased as MCR decreased (P < 0.05). In the challenged group (CG), body weight and lean tissue weight were unaffected by MCR, and BMC linearly increased as MCR decreased (P < 0.05). For the cortical bone of femoral metaphysis on 6 DPI, bone mineral density (BMD) linearly increased as MCR decreased (P < 0.05). Bone volume to tissue volume ratio (BV/TV) in the CG linearly increased as MCR decreased (P < 0.05). On 9 DPI, BMC and TV linearly increased as MCR decreased (P < 0.05) in the NCG. BMD and BV/TV changed quadratically as MCR decreased (P < 0.05). For the trabecular bone of femoral metaphysis on 9 DPI, BV/TV, and trabecular number linearly increased as MCR decreased (P < 0.05) in the NCG. For the femoral diaphysis, BV, TV, BMC on 6 DPI, and BMD on 9 DPI linearly increased as MCR decreased (P < 0.05). In conclusion, this study showed that both Eimeria challenge and varying supplemental MCR could influence bone quality of broilers.

Steroid-Insensitive Gene Expression of Extracellular Matrix Components and Pro-fibrotic Factors in the Lung Associated with Airway Hyperresponsiveness in Murine Asthma.

Between 5 and 10% of asthma patients do not respond to glucocorticoid therapy. Experimental animal models are indispensable for investigating the pathogenesis of steroid-resistant asthma; however, the majority of murine asthma models respond well to glucocorticoids. We previously reported that multiple intratracheal administration of ovalbumin (OVA) at a high dose (500 µg/animal) induced steroid-insensitive airway eosinophilia and remodeling with lung fibrosis, whereas a low dose (5 µg/animal) caused steroid-sensitive responses. The aims of the present study were as follows: 1) to clarify whether airway hyperresponsiveness (AHR) in the two models is also insensitive and sensitive to a glucocorticoid, respectively, and 2) to identify steroid-insensitive genes encoding extracellular matrix (ECM) components and pro-fibrotic factors in the lung. In comparisons with non-challenged group, the 5- and 500-µg OVA groups both exhibited AHR to methacholine. Daily intraperitoneal treatment with dexamethasone (1 mg/kg) significantly suppressed the development of AHR in the 5-µg OVA group, but not in the 500-µg OVA group. Among genes encoding ECM components and pro-fibrotic factors, increased gene expressions of fibronectin and collagen types I, III, and IV as ECM components as well as 7 matrix metalloproteinases, tissue inhibitor of metalloproteinase-1, transforming growth factor-β1, and activin A/B as pro-fibrotic factors were insensitive to dexamethasone in the 500-µg OVA group, but were sensitive in the 5-µg OVA group. In conclusion, steroid-insensitive AHR developed in the 500-µg OVA group and steroid-insensitive genes encoding ECM components and pro-fibrotic factors were identified. Drugs targeting these molecules have potential in the treatment of steroid-resistant asthma.

Graded levels of Eimeria infection modulated gut physiology and temporarily ceased the egg production of laying hens at peak production

An experiment was conducted to investigate the changes in gastrointestinal physiology, including intestinal leakage, immune response, oxidative stress, along with performance traits, of Hy-Line W-36 laying hens following Eimeria infection at peak egg production. A total of 360 laying hens, at 25 wk of age, were assigned randomly to 5 treatment groups, each consisting of 6 replicate cages, including a nonchallenged control group. The other 4 groups were inoculated with graded levels of mixed Eimeria species as high, medium-high, medium-low, and low doses, respectively. The body weight (BW) and body weight gain (BWG) of laying hens were measured from 0 to 14 days postinoculation (DPI). Average daily feed intake (ADFI) and hen-day egg production (HDEP) were measured from 0 to 15 and 0 to 28 DPI, respectively. Gut permeability was measured on 5 DPI, whereas oxidative stress, immune response, and expression of nutrient transporter genes were measured on 6 and 14 DPI. A significant linear reduction in BW and BWG was observed with increasedEimeriainoculation dosage on both 6 and 14 DPI (P < 0.001,P-Lin < 0.0001). An interaction between theEimeriadosages and DPI was observed for ADFI (P < 0.0001). Feed intake in the challenged groups decreased starting at 4 DPI, with the most significant drop occurring at 7 DPI, which did not recover until 15 DPI. Following the challenge, gastrointestinal physiology shifted toward the host defense against the Eimeria infection by upregulating mRNA expression of tight junction proteins and immune responses while downregulating the expression of key nutrient transporters on 6 and 14 DPI (P < 0.05). An interaction between theEimeriainoculation dosage and DPI was also observed for daily HDEP (P < 0.0001). Overall, HDEP was lower in the challenged groups compared to the control. Daily HDEP in the challenged groups dropped from 8 DPI and became similar to the control birds only after 24 DPI. Egg production temporarily ceased in most of the laying hens infected with the high and medium-high dosages of Eimeria. In conclusion, Eimeria infection activated the host immune response, negatively affecting the gastrointestinal health, growth performance, and temporarily ceased the egg production of Hy-Line W-36 laying hens when infected at peak production.

Buffered formic acid and a monoglyceride blend improve performance and modulate gut bacteria and immunity gene expression in broilers under necrotic enteritis challenge

Due to the removal of antibiotics from animal feed, alternatives have been sought to control necrotic enteritis (NE) in broilers. The current study investigated the effects of buffered formic acid (Amasil NA) and monoglycerides of short- and medium-chain fatty acids (Balangut LS P) on the performance and gut health of broilers challenged with NE. A total of 816 as-hatched 1-d-old chicks (Cobb 500) were randomly assigned to 6 treatments with 8 replicates. Treatments were: T1) nonchallenged control; T2) NE challenged control; T3) Amasil NA (challenge plus Amasil NA, 0.3% throughout all phases); T4) Balangut LS P (challenge plus Balangut LS P, 0.5%, 0.3%, and 0.2% in the starter, grower and finisher phases, respectively; T5) Combined (challenge plus combination of T3 and T4); T6) Antibiotic (challenge plus Zn bacitracin, 0.05 % throughout all phases). Birds were orally gavaged with live Eimeria vaccine species (d 9) and with Clostridium perfringens (d 14 and 15). On d 16, birds were sampled to evaluate gut permeability, microbiota, and mRNA abundance in the jejunum. The data were analyzed in JMP software using one-way ANOVA with Tukey's test to separate means, and Kruskal-Wallis test was used for non-normally-distributed parameters. Results showed that Balangut LS P decreased (P<0.05) feed conversion ratio compared to nonchallenged ones at the end of the study. Balangut LS P reduced (P < 0.05) the level of cecal Bacteriods compared to nonchallenged group, whereas Amasil NA shifted the levels of ileal Bifidobacteria, Enterobacteriaceae, and Lactobacillus towards nonchallenged control (P > 0.05). NE challenge upregulated (P < 0.001) the expression of IL-21R, zeta chain of T cell receptor (ZAP70), and dual specificity phosphatase 4 (DUSP4) compared to nonchallenged birds, whereas Balangut LS P showed an intermediate (P > 0.05) expression pattern of these genes towards nonchallenged and antibiotic groups. In conclusion, combination of Balangut LS P and Amasil NA has the potential to be used as an additive to improve the performance and gut health of broiler chickens, especially under challenging conditions such as NE infections.

Effects of stimbiotic supplementation on gut health, immune response, and intestinal microbiota in weaned piglets challenged with E. coli.

In order to make piglet diets more effective, it is necessary to investigate effective methods for breaking down xylan in cereal. The objective of this study was to determine the effects of dietary stimbiotic (STB) supplementation on growth performance, intestinal morphology, immune response and intestinal microbiota in weaned piglets. A total of 24 (Duroc × Yorkshire × Landrace) weaned pigs (initial body weight of 8.01 ± 0.38 kg and 28 ± 3 d old), were assigned to 4 treatments with 6 replicates per treatment. Pigs were housed in individual pens for 17 days, including 5 days adaption period and 12 days after the first Escherichia coli (E. coli) challenge. The experiment was conducted in a 2 × 2 factorial arrangement of treatments consisting of two levels of challenge (challenge and non-challenge) and two levels of STB (0 and 0.5 g/kg diet). Supplementations of STB 0.5 g/kg improved the gain to feed ratio (G:F) (P < 0.05) in piglets challenged with shiga toxigenic E. coli (STEC). STB supplementation decreased (P < 0.05) white blood cells, neutrophils, lymphocytes, and expression levels of tumor necrosis factor-alpha and interleukin-6. Supplementation of STB improved (P < 0.05) the lymphocytes and neutrophils in piglets challenged with STEC on 12 dpi. Supplementation of STB also improved (P < 0.05) the villus height to-crypt depth ratio of ileum in piglets challenged with STEC. Supplementation of STB increased (P < 0.05) the expression levels of claudin-1 of ileum. In genus level, supplementation of STB increased (P < 0.001) the abundance of Prevotella compared to non-supplementation of STB groups in pre-inoculation period. Also, supplementation of STB decreased (P < 0.05) the abundance of Faecalibacterium and Eubacterium_coprostanoligenes_group compared to non-supplementation of STB groups in post-inoculation period. In phylum level, supplementation of STB increased (P < 0.05) the abundance of Desulfobacterota and Fibrobacterota in pre-inoculation period. E. coli challenge increased the abundance of Fibrobacterota compared to non-challenged group in post-inoculation period. In conclusion, these findings indicated that STB supplementation could alleviate a decrease of the performance, immune response, and inflammatory response in piglets induced by the STEC challenge.

Applying different morphometric intestinal mucosa methods and the correlation with broilers performance under Eimeria challenge

The intestinal wall has on its surface, protrusions called villi that are responsible for the absorption of nutrients. Commonly, these structures have their dimensions measured to related more area surface with better absorption. However, the measurement of these villi neglects the inflammation and the presence of immature cells that increase the surface area but affect negatively the absorption and compromise the animal performance. The measurements of villi/crypt are traditional tools in animal research; however, they may overlook alterations that impact the mucosal functionality. This study aimed to compare the morphometry of the intestinal villi/crypt with the I See Inside (ISI) scoring methodology, exploring their correlation with zootechnical performance. Therefore, broilers were grouped as nonchallenged (NC) and challenged with Eimeria (CH) and jejunum samples were collected at 22 d for histological analysis. The same villi were submitted to the ISI methodology, which is based on the scoring of 8 parameters related to the inflammatory process, and the measurements of villus height (VH), villus width (VW), crypt depth (CD), crypt width (CW), VH:CD ratio and villi absorptive surface (VAS). The CH group presented higher ISI total score, VW, CD, CW and lower VH, VH:CD, and VAS in comparison to the NC group. While the villi/crypt morphometry did not exhibit correlations with performance, the presence of Eimeria oocysts and the ISI total score was positively correlated (P < 0.05) with the feed conversion ratio (FCR), demonstrating a statistical interaction between high ISI scores and worse performance. In conclusion, a larger villus is not related to better intestinal functionality when this enlargement is unleashed by the immune processes occurring inside. The scoring system that evaluates the type of alteration observed has a direct impact on the animal's zootechnical performance which is not observed with the single metric surface evaluation.

The effects of probiotic and threonine application on the carcass yield, internal organ development, intestinal morphology and cecal microbiota of broilers challenged with Clostridium perfringens

The aim of this study was to explore the effects of probiotics (Ecobiol®) and threonine supplements on broiler internal organs and intestinal health under Clostridium perfringens challenge. A total of 1600 male Ross 308 broiler chicks were randomly assigned to eight treatments with eight replicates each of 25 birds. Dietary treatments consisted of two levels of supplemented threonine (without and with threonine supplementation), two levels of probiotics (Ecobiol®) supplement (0 and 0. 1% of diet), and two levels of challenge (without and with 1 ml of the C. perfringens inoculum (∼108 cfu/ml) on d 14, 15, and 16 of the experiment), which fed to the birds during a 42 d feeding trial. The results showed that adding threonine and probiotic supplements to the diets of C. perfringens-infected birds reduced the relative gizzard weight by 22.9% compared to those fed un-supplemented diet (P ≤ 0.024). As compared to the non-challenged group, the C. perfringens challenge significantly reduced the carcass yield of broilers by 1.18% (P < 0.0004). The groups receiving threonine and probiotic supplementation had higher carcass yield, and the inclusion of probiotics in the diet decreased abdominal fat by 16.18% compared with the control treatment (P ≤ 0.001). Adding threonine and probiotic supplements to the diets of broilers challenged with C. perfringens increased the jejunum villus height in comparison with C. perfringens-infected group fed an unsupplemented diet on day 18 (P ≤ 0.019). The number of cecal E. coli increased in birds under C. perfringens challenge in comparison with the negative group. Based on the findings, dietary inclusion of threonine and probiotic supplement could beneficially affect intestine health and carcass weight during the C. perfringens challenge.

Potential of cinnamaldehyde essential oil as a possible antimicrobial against fowl typhoid in layers.

Fowl typhoid (FT) is an economically significant bacterial disease of layers leading to a drastic drop in egg production. Due to increased public health concerns about antibiotics in poultry feed, a search for new safe antimicrobials for treating fowl typhoid is crucial. The antimicrobial effect of cinnamaldehyde essential oil (CnEO) against fowl typhoid in layers was investigated in this experiment. The 60-week-old BV300-layer birds (n = 100) were divided into five groups: the non-challenged control group A, only cinnamaldehyde-treated group B (CnEO @ 1:8000 dilutions through drinking water for 60days), the challenged group C, challenged plus cinnamaldehyde therapy group D (CnEO @ 1:8000 dilutions through drinking water from 16 to 30 dpi), and challenged plus antibiotic therapy group E (chloramphenicol @ 1 gm/5lit through drinking water from 16 to 30 dpi). Hens from all challenged groups were challenged with Salmonella Gallinarum (VTCCBAA588) @ 1 × 108CFU/ml orally. Various parameters such as clinical signs, mortality, egg production and egg weight, colony-forming unit (CFU) count of cecal content, eggshell surface, and egg yolk were evaluated all through 60days of an experimental trial. Results indicated that, in the case of the cinnamaldehyde therapeutic group, there was a significant improvement in egg production, mild clinical signs, lower feed conversion ratio (FCR), and a significantly lower bacterial count in ceca and on the eggshell surface compared to the control challenge group. Thus, CnEO @ 1:8000 dilutions through drinking water can be a potential antimicrobial for controlling fowl typhoid.

Time effect of experimental infection on Rainbow trout (Oncorhynchus mykiss) by immersion with Aeromonas salmonicida subsp. salmonicida

Furunculosis caused by Aeromonas salmonicida subsp salmonicida (Ass) is a medically and economically important bacterial disease in salmonid farms that requires therapeutic measures to prevent and control the disease. Evaluation of the effectiveness of traditional measures such as antibiotics or vaccines usually requires infecting fish experimentally. The objective of this study is to develop a method of infectious challenge of large (250-g) Rainbow trout by immersion close to natural infection conditions. We compare mortality, morbidity and anti-Ass antibody production of Rainbow trout following different bathing times (2, 4, 8 and 24 h) at a final bacterial concentration of 106 CFU/mL. One hundred sixty fish divided in five groups corresponding to the 4 bathing times and the non-challenged group were studied. The 24 h contact duration resulted in the infection of all fish, with a mortality rate of 53.25%. The challenged fish developed acute infection with symptoms and lesions (inappetance, altering of swimming behaviour, presence of boils) similar to those observed in furunculosis, and produced antibodies against the bacterium at 4 weeks after challenging, in contrast with the non-challenged group.

160 Effects of Bacillus Licheniformis PF9 on Growth, Nutrient Absorption, gut Barrier Integrity, Immunity, and Intestinal Microbiota of Weaned Piglets Challenged with Enterotoxigenic Escherichia Coli F4

Abstract This study aimed to evaluate the effects of Bacillus licheniformis PF9 on growth performance and gut health of weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC) F4. Thirty-two weaned piglets susceptible of ETEC F4 were randomly allocated to 4 treatments 1) NNC, basal diet and non-challenged; 2) NC, basal diet; 3) PC, basal diet + 80 mg·kg-1 of Avilamycin; and 4) PF9, basal diet + 109 cfu/kg of Bacillus licheniformis PF9. ETEC F4 decreased average daily gain and increased feed conversion ratio of the NC group. An increase was found in FCR of the PC group during the post-challenge period (P &amp;lt; 0.05). ETEC F4 induced the severity of diarrhea at 3-, 36- and 40-hours post-inoculum (hpi) in the NC group (P &amp;lt; 0.05). Bacillus licheniformis relieved diarrhea severity at 3 hpi (P &amp;lt; 0.05). ETEC F4 reduced the duodenal, jejunal and ileal villus height of the NC group. A decrease in duodenal villus height in the PC group (P &amp;lt; 0.05). ETEC F4 reduced the relative mRNA level of Na+-glucose cotransporter 1 in the NC group (P &amp;lt; 0.05). Compared to the NC and NNC groups, Bacillus licheniformis increased relative mRNA levels of Aminopeptidase N, Occludin, Zonula occludens-1, and SGLT1 (P &amp;lt; 0.05). Bacillus licheniformis increased the relative mRNA level of excitatory amino acid transporter 1 (P &amp;lt; 0.05). Bacillus licheniformis showed a lower relative abundance of Bacteroidetes in the colon than piglets from the NNC group (P &amp;lt; 0.05). The non-challenged group had a higher relative abundance of Firmicutes in the ileum than the challenged piglets, meanwhile, a lower relative abundance of Proteobacteria in the ileum and colon in the NC group (P &amp;lt; 0.05). This study provides evidence that Bacillus licheniformis can be a potential antibiotics alternative to improve the gut health of piglets.

Chicken Gut Microbiota Responses to Dietary Bacillus subtilis Probiotic in the Presence and Absence of Eimeria Infection.

Coccidiosis is a well-known poultry disease that causes the severe destruction of the intestinal tract, resulting in reduced growth performance and immunity, disrupted gut homeostasis and perturbed gut microbiota. Supplementation of probiotics were explored to play a key role in improving growth performance, enhancing innate and adaptive immunity, maintaining gut homeostasis and modulating gut microbiota during enteric infection. This study was therefore designed to investigate the chicken gut whole microbiota responses to Bacillus subtilis (B. subtilis) probiotic feeding in the presence as well as absence of Eimeria infection. For that purpose, 84 newly hatched chicks were assigned into four groups, including (1) non-treated non-challenged control group (CG − ET), (2) non-treated challenged control group (CG + ET), (3) B. subtilis-fed non-challenged group (BS − ET) and (4) B. subtilis-fed challenged group (BS + ET). CG + ET and BS + ET groups were challenged with Eimeria tenella (E. tenella) on 21 day of housing. Our results for Alpha diversity revealed that chickens in both infected groups (CG + ET and BS + ET) had lowest indexes of Ace, Chao 1 and Shannon, while highest indexes of Simpson were found in comparison to non-challenged groups (CG − ET and BS − ET). Firmicutes was the most affected phylum in all experimental groups following Proteobacteria and Bacteroidota, which showed increased abundance in both non-challenged groups, whereas Proteobacteria and Bacteroidota affected both challenged groups. The linear discriminant analysis effect size method (lEfSe) analysis revealed that compared to the CG + ET group, supplementation of probiotic in the presence of Eimeria infection increased the abundance of some commensal genera, included Clostridium sensu stricto 1, Corynebacterium, Enterococcus, Romboutsia, Subdoligranulum, Bacillus, Turicibacter and Weissella, with roles in butyrate production, anti-inflammation, metabolic reactions and the modulation of protective pathways against pathogens. Collectively, these findings evidenced that supplementation of B. subtilis probiotic was positively influenced with commensal genera, thereby alleviating the Eimeria-induced intestinal disruption.

Effects of mixed Eimeria challenge on performance, body composition, intestinal health, and expression of nutrient transporter genes of Hy-Line W-36 pullets (0-6 wks of age)

A study was aimed to investigate the effects of mixed Eimeria challenge on performance, gastrointestinal health, oxidative stress, inflammation, and expression of nutrient transporter genes of Hy-Line W-36 pullets. A total of 540, 16-d old pullets were randomly allocated into 5 treatment groups with 6 replicate cages, including a nonchallenged control group. A mixed Eimeria species solution containing 50,000 E. maxima, 50,000 E. tenella, and 250,000 E. acervulina oocysts per mL was prepared and challenged to one group as a high-dose treatment (High). The 2-fold serial dilution was done to prepare the medium-high (Med-High: 25,000 E. maxima; 25,000 E. tenella; and 125,000 E. acervulina), the medium-low (Med-Low: 12,500 E. maxima; 12,500 E. tenella; and 62,500 E. acervulina), and the low (Low: 6,250 E. maxima; 6,250 E. tenella; and 31,250 E. acervulina) dose treatments, and these dosages were challenged to 3 remaining groups, respectively. Growth performance, daily feed intake (FI), and mortality were calculated from 0-14 d postinfection (DPI). Gastrointestinal permeability (GP) was measured on 3, 5, 6, 7, and 9 DPI. The result indicated significant linear responses to the Eimeria challenge dosage in average body weight and body weight gain (P < 0.0001). An interaction between treatment and DPI was observed for FI (P < 0.0001). Feed intake significantly dropped from 4 DPI and did not recover until 12 DPI in the challenged groups. The lowest FI for each of the challenged groups was observed on 5 DPI. Gastrointestinal permeability increased linearly, peaking at 5 DPI, and was recovered back to normal by 9 DPI in the challenged groups. Furthermore, gene expression of tight junction proteins was linearly upregulated by increased Eimeria dosages. The oxidative status of the pullets was lowered in the challenged groups than the nonchallenged control group, whereas the expression of inflammatory and proinflammatory cytokines was upregulated by Eimeria challenge on 6 DPI (P < 0.05). The highest mortality was observed in pullets challenged with the High, followed by the Med-High (P < 0.0001) on 5 DPI. In summary, the mixed Eimeria challenge linearly reduced the growth performance of pullets with an increase in oxidative stress and inflammation. A severe effect of Eimeria on gastrointestinal health was observed on 5 or 6 DPI as suggested by GP, tight junction genes, and mortality results. This study indicates that Eimeria infection can be a threat to gastrointestinal health related issues in pullets.

PSII-16 Enterid Reduced E. Coli Colonization in the Gastro-Intestinal Tract of Young Pigs

Abstract The aim of the present study is to assess the impact of Enterid feed additive (PMI Additives) on the health status of newly-weaned pigs challenged with Enterotoxigenic E. coli (F4/K88). Seventy-two newly-weaned piglets of mixed sex were weighed and sorted into groups of 12 pens (3 gilts and 3 barrows each pen). Each pen was then randomly assigned into 1 of 3 treatments, consisting of a non-challenged group (NC), E.coli-challenged group (ETEC), and ETEC supplemented with 0.10% Enterid. Pigs were treated with Baytril upon arrival and were provided their respective diets and water ad libitum. All pigs except NC were challenged with F4/K88 strain 181016 E. coli on d 10 and d 11. Clinical observations (fecal scores, respiration rate, appetite, attitude, edema), rectal temperature and E. coli shedding were measured daily beginning on d9 until the end of the study (d 20). Clinical outcomes were analyzed using Mixed procedure in SAS following a completely randomized design with initial weight as covariate and day post challenge as a repeated measure parameter. Clinical measurements, except fecal scores, were all rated normal and so were not analyzed statistically. There were no differences among treatments in E. coli shedding prior to the challenge (2.3 log cfu/ml, P &amp;gt; 0.20). Immediately post challenge, however, E. coli shedding for the ETEC increased by 12 log (14.4 log cfu/ml, P &amp;lt; 0.05) and remained elevated for 4-5 d. Supplementation with Enterid resulted in a 2-log reduction in E. coli shedding during the first 3 d post-challenge (P &amp;lt; 0.10) and returned to pre-challenge level after 5 d. No clear differences in rectal temperature were observed among treatments. E. coli challenge resulted in poorer fecal quality (d3-5, P &amp;lt; 0.05), with Enterid not influencing the response (P &amp;gt; 0.20). The current results support the ability of Enterid to reduce E. coli colonization in the gut of pigs challenged with E. coli.

Development of an environmental contamination model to simulate the microbial bloom that occurs in commercial hatch cabinets

Microbial blooms that emerge in commercial hatch cabinets consist of apathogenic and pathogenic microorganisms, including Escherichia coli, Enterococcus faecalis, and Aspergillus fumigatus. Objectives of the present study included the development of a multipathogen contamination model to mimic commercial conditions and optimization of sampling methods to quantify bacterial or fungal presence within the hatch cabinet. The pathogen challenge mix (PM) was recreated from select bacterial or fungal isolates recovered from an egg homogenate (EH) derived from the contents of infertile eggs and late embryonic mortalities. Isolates selected for PM included Enterococcus faecalis (∼108 CFU/egg), Staphylococcus aureus (∼107 CFU/egg), Staphylococcus chromogenes (∼107 CFU/egg), Aspergillus fumigatus (∼106 spores/egg), and 2 Escherichia coli (∼108 CFU/egg) isolates. Challenge (100 μL of PM or EH) was administered using a sterile loop to a 28 mm area on the blunt end of the eggshell at day 19 of embryogenesis (DOE). In 3 experiments, microbiological data were collected from environmental hatcher samples (open-agar plate method), fluff samples, postmortem whole-body chick rinse samples, and gastrointestinal tract (GIT) samples to evaluate select bacteria and fungi circulating within the hatch cabinet and colonization of GIT. Cumulative bacterial and fungal recovery from the PM hatching environment from DOE20 to hatch was higher than the nonchallenged group (NC) and EH group at ∼860 and ∼1,730 CFU, respectively. Bacterial recovery from GIT, fluff, and chick rinse samples were similar for the PM and EH group in Exp. 1. However, Aspergillus fumigatus recovery from fluff and chick rinse samples for the PM group was significantly (P < 0.001) higher than the NC and EH group. In Exp. 2 and 3, PM challenge significantly (P < 0.05) increased Gram-negative bacterial recovery from the GIT, fluff and chick rinse samples compared to both the NC and EH group. These data suggest this innovative multispecies environmental contamination model using PM could be utilized to evaluate strategies to mitigate microbial contamination in commercial hatch cabinets in a laboratory setting.

Dietary gamma-aminobutyric acid supplementation does not mitigate stress responses in weaner pigs given adrenocorticotropic hormone and experimentally infected with enterotoxigenic Escherichia coli

Gamma-aminobutyric acid (GABA) is a non-protein amino acid, a major inhibitory neurotransmitter in the central nervous system that stimulates feed intake and inhibits the hypothalamic-pituitary-adrenal (HPA) axis. We hypothesised that: (1) GABA supplementation in the diet would reduce markers of the stress response in weaned pigs injected with adrenocorticotropic hormone (ACTH) and inoculated with enterotoxigenic Escherichia coli (ETEC); and (2) this reduction in stress responses would improve performance after weaning. A total of 96 newly-weaned male pigs (Large White x Landrace) were stratified into a two by four factorial arrangement with respective factors being (i) with/without ACTH injection and ETEC infection (challenge versus non-challenge) and (ii) four dietary GABA levels (0, 60, 80, 100 mg/kg). On days 8 and 9 after weaning, piglets were orally inoculated with ETEC (0.8 ml via two gelatinized capsules; serotype O149; F4) as well as being given 5 IU ACTH intramuscularly (IM), which occurred an hour before ETEC inoculation. Pigs in the non-challenged group were given IM 0.2 mL of sterile saline and sham infected with two capsules of PBS. Faecal consistency scores were recorded daily, pigs and feed were weighed weekly to determine performance and blood samples were collected at days 6, 9 and 14. GABA supplementation did not reduce plasma cortisol. However challenged pigs had higher levels compared to the non-challenge group at day 9 (P = 0.001). Performance was not influenced by GABA supplementation (P > 0.05). Between days 8–14, 54.3% of pigs in the challenge group developed diarrhoea compared to 5.6% in the non-challenged group (P = 0.001). These data indicate that eliciting both an ETEC infection challenge and an acute stress response after weaning initiated an endocrine stress response. The use of GABA in feed did not reduce this stress response, reduce diarrhoea or improve production performance.

Effect of organic acids on growth performance, intestinal morphology, and immunity of broiler chickens with and without coccidial challenge

A total of 360-day-old broiler chicks were allocated into six groups in 2 (Coccidial challenge or not) × 3 (dietary treatments) factorial design. Three dietary treatments including: basic diet, basic diet plus organic acids (OAs) in drinking water, and basic diet plus OAs in the feed with and without coccidial challenge. The OAs in water or feed improved (P < 0.01) average body weight (ABW), average body weight gain (ABWG), and feed conversion ratio (FCR) as compared with the control diet during starter, grower, and whole experimental period. Coccidial challenge decreased BW, ABWG, and average feed intake (AFI), as well as resulted in poor FCR during the starter and whole experimental period (P < 0.05). Though there was no interaction between OAs supplementation and coccidial challenge, the OAs supplementation improved the overall performance with and without coccidial challenge birds on 21 d and 35 d. IgG was found higher (P = 0.03) in broilers fed OAs in feed without the coccidial challenge group. On 18 d, OAs supplementation in feed increased TNF-γ (P = 0.006), whereas the coccidial challenge decreases TNF-γ (P = 0.01) and IL-10 (P = < .0001), and increases IgM (P = 0.03), IgG (P = 0.04) and IgA (P = 0.02). On 29 d, the coccidial challenge increases IgM and IgA. On 18 d, jejunal lesion score was found significantly higher in the coccidial challenged group as compared to OAs supplementation with coccidial challenged groups on 18 d (P < 0.0001) and 29 d (P = 0.03). Crypt depth was higher, and Villus-height to Crypt depth ratio was lower in the coccidial challenge group on 18 and 29 d. The Goblet cells were found higher in the non-coccidial challenge on 18 d. After 18 d, 16S rDNA gene sequence analysis of ileal chyme has shown that coccidial challenge decreases Lactobacillus_reuteri species as compared to the non-challenged group (P = 0.02). After 29, Cyanobacteria abundance reduced (P = 0.014) in the challenged group than the non-challenged group at the phylum level. At the genus level, Lactobacillus (P = 0.036) and unidentified Cyanobacteria (P = 0.01) were found higher in the non-challenged group than the coccidial challenge group. The results indicate that the OAs supplementation showed improved responses in a pattern similar to the non-challenged control group by neutralizing the negative effects of the coccidial challenge.

Resveratrol protects intestinal integrity, alleviates intestinal inflammation and oxidative stress by modulating AhR/Nrf2 pathways in weaned piglets challenged with diquat

This study investigated the effects of resveratrol (RES) on intestinal morphology, antioxidant capacity, intestinal inflammation, and barrier function in weaned piglets challenged with diquat (DIQ). Thirty weaned piglets were randomly assigned to 5 treatments: non-challenged group (CON), DIQ-challenged group (DIQ), and DIQ-challenged group with 10, 30, or 90 mg/kg of RES, respectively. The trail lasted 21 days, and piglets were intraperitoneally injected with DIQ or the same amount of saline on day 15. The results showed that supplementation with 90 mg/kg RES increased (P < 0.05) jejunal villus height and villus height: crypt depth ratio, and decreased (P < 0.05) crypt depth, plasma D-lactate and diamine oxidase (DAO) compared with the DIQ group. Piglets fed with 30 or 90 mg/kg RES prevented the diquat-induced decrease (P < 0.05) of mRNA expression of occludin, claudin-1, ZO-1, and IL-10, and increase (P < 0.05) of TNF-α mRNA expression. Moreover, addition of 90 mg/kg RES increased (P < 0.05) the activities of SOD, GSH-Px, and CAT and decreased (P < 0.05) the MDA levels in jejunal mucosa compared with the DIQ group. Finally, addition of 90 mg/kg RES enhanced (P < 0.05) the mRNA expression of SOD1, SOD2, CAT, GPx1, and HO-1, and increased (P < 0.05) mRNA and protein expression of Nrf2, NQO1, aryl hydrocarbon receptor (AhR), and cytochrome P450 family 1 member A1 (CYP1A1). These data indicated that supplementation with 90 mg/kg RES was effective in protecting the intestinal integrity, alleviating intestinal inflammation and oxidative stress by activating AhR/Nrf2 pathways in diquat-challenged piglets.