Two intestinal microbiota-derived metabolites, deoxycholic acid and butyrate, synergize to enhance host defense peptide synthesis and alleviate necrotic enteritis

Background Necrotic enteritis(NE)is a major enteric disease in poultry,yet effective mitigation strategies remain elusive.Deoxycholic acid(DCA)and butyrate,two major metabolites derived from the intestinal microbiota,have independently been shown to induce host defense peptide(HDP)synthesis.However,the potential synergy between these two compounds remains unexplored.Methods To investigate the possible synergistic effect between DCA and butyrate in regulating HDP synthesis and barrier function,we treated chicken HD11 macrophage cells and jejunal explants with DCA and sodium butyrate(NaB),either individually or in combination,for 24 h.Subsequently,we performed RNA isolation and reverse transcrip-tion-quantitative PCR to analyze HDP genes as well as the major genes associated with barrier function.To further determine the synergy between DCA and NaB in enhancing NE resistance,we conducted two independent trials with Cobb broiler chicks.In each trial,the diet was supplemented with DCA or NaB on the day-of-hatch,followed by NE induction through sequential challenges with Eimeria maxima and Clostridium perfringens on d 10 and 14,respectively.We recorded animal mortality after infection and assessed intestinal lesions on d 17.The impact of DCA and NaB on the microbiota in the ileum and cecum was evaluated through bacterial 16S rRNA gene sequencing.Results We found that the combination of DCA and NaB synergistically induced multiple HDP genes in both chicken HD11 cells and jejunal explants.Additionally,the gene for claudin-1,a major tight junction protein,also exhibited synergistic induction in response to DCA and NaB.Furthermore,dietary supplementation with a combination of 0.75 g/kg DCA and 1 g/kg NaB led to a significant improvement in animal survival and a reduction in intestinal lesions compared to either compound alone in a chicken model of NE.Notably,the cecal microbiota of NE-infected chickens showed a marked decrease in SCFA-producing bacteria such as Bacteroides,Faecalibacterium,and Cuneatibacter,with lactobacilli becoming the most dominant species.However,supplementation with DCA and NaB largely restored the intestinal microbiota to healthy levels.Conclusions DCA synergizes with NaB to induce HDP and claudin-1 expression and enhance NE resistance,with potential for further development as cost-effective antibiotic alternatives.

Dietary Macleaya cordata extract supplementation improves the growth performance and gut health of broiler chickens with necrotic enteritis

Background The poultry industry needs effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE)caused by Clostridium perfringens.Methods The aim of this study was to investigate the effects of dietary supplementation with Macleaya cordata extract(MCE)on the immune function and gut microbiota of broilers with NE.A total of 2881-day-old broiler chicks were randomly assigned to a 2×2 factorial arrangement with two concentrations of dietary MCE supplementation(0 or 350 mg/kg of diet)and two disease challenge statuses(control or NE).Results The results revealed that NE significantly increased the feed conversion rate(FCR),mortality,intestinal lesion score,the levels of IL-1β,IL-17 and IFN-γ/IL-4 in serum and IL-17/IL-10 in the jejunal mucosa,m RNA levels of TLR2,IFN-γand p Ig R in the jejunum,and Clostridium perfringens concentrations in the cecum.NE significantly decreased the body weight(BW),body weight gain(BWG),jejunal villus height,V/C,m RNA level of AMPK-α1 in jejunum,IL-4 level in the jejunal mucosa and lactic acid bacteria abundance in the cecum.MCE significantly increased BW,BWG,jejunal villus height,V/C,m RNA levels of occludin,ZO-1 and AMPK-α1 in the jejunum,the levels of Ig A and Ig G in serum and IL-10 in the jejunal mucosa and m RNA levels of NF-κB,IL-10 and MHC-II in the jejunum.Additionally,MCE significantly decreased the FCR,mortality,intestinal lesion score,jejunal crypt depth,the levels of IFN-γand IL-17 in serum and IL-17/IL-10 in the jejunal mucosa,Clostridium perfringens concentrations in the cecum,and m RNA levels of IL-17/IL-10 in the jejunum.Moreover,NE significantly increased the abundance of bacteria that are associated with inflammation,obesity and depression(Alistipes,Barnesiella,Intestinimonas,RF39 and UCG-005)and significantly decreased the abundance of short-chain fatty acid(SCFA)-producing bacteria(Anaerotruncus,Butyricicoccus and Bacteroides)in the cecum.MCE significantly increased the abundance of SCFA-producing bacteria(Streptococcus,Ruminococcus_torques_group and Lachnospiraceae_NK4A136_group)and significantly reduced the abundance of bacteria that are associated with inflammation and obesity(Alistipes,Barnesiella and UCG-010)in the cecum.In the cecum of broilers with NE,the relative abundance of Barnesiella and Alistipes was higher and that of Lachnoclostridium and Shuttleworthia was lower.Interestingly,these trends were reversed by the addition of MCE to the diet.Spearman correlation analysis showed that Barnesiella and Alistipes were associated with enhanced intestinal inflammation and inhibited growth performance,whereas Lachnoclostridium and Shuttleworthia were associated with anti-inflammatory effects.Conclusions MCE ameliorated the loss of growth performance in broiler chickens with NE,probably by regulating the intestinal barrier,immune function,and gut microbiota.

Intestinal microecological transplantation for a patient with chronic radiation enteritis:A case report

BACKGROUND The gut microbiota is strongly associated with radiation-induced gut damage.This study aimed to assess the effectiveness and safety of intestinal microecological transplantation for treating patients with chronic radiation enteritis.CASE SUMMARY A 64-year-old female with cervical cancer developed abdominal pain,diarrhea,and blood in the stool 1 year after radiotherapy.An electronic colonoscopy was performed to diagnose chronic radiation enteritis.Two courses of intestinal microecological transplantation and full-length 16S rRNA microbiological analysis were performed.The patient experienced short-and long-term relief from symptoms without adverse effects.Whole 16S rRNA sequencing revealed significant differences in the intestinal flora’s composition between patient and healthy donors.Pathogenic bacteria,such as Escherichia fergusonii and Romboutsia timonensis,were more in the patient.Beneficial bacteria such as Faecalibacterium prausnitzii,Fusicatenibacter saccharivorans,Ruminococcus bromii,and Bifidobacterium longum were more in the healthy donors.Intestinal microbiota transplantation resulted in a significant change in the patient's intestinal flora composition.The composition converged with the donor's flora,with an increase in core beneficial intestinal bacteria,such as Eubacterium rectale,and a decrease in pathogenic bacteria.Changes in the intestinal flora corresponded with the patients'alleviating clinical symptoms.CONCLUSION Intestinal microecological transplantation is an effective treatment for relieving the clinical symptoms of chronic radiation enteritis by altering the composition of the intestinal flora.This study provides a new approach for treating patients with chronic radiation enteritis.

Potential pharmacological mechanisms of digallate in the treatment of enteritis based on network pharmacology and molecular docking

Background:In order to investigate the possible pharmacological mechanism of digallate in Galla Chinensis for treating enteritis,providing reference for the search and exploration of effective drugs for treating enteritis.Method:Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform,PharmMapper,DisGeNET,DrugBank,and GeneCards databases were used to obtain drug and disease-related target information.Gene ontology functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment were performed,and the main therapeutic pathways and targets were identified by combining protein-protein interaction networks and cytoHubba plug-in.Molecular docking was used to validate the results.Result:297 drug related targets,2436 disease related targets,and 66 target points related to digallate were predicted to be associated with enteritis.10 related signal pathways and 10 key genes were identified.Conclusion:Digallate may be utilized to treat enteritis by acting on similar pathways,such those related to pathways in cancer,lipid and atherosclerosis,proteoglycans in cancer,Rap1 signaling pathway,PI3K-Akt signaling pathway and other targets such as IGF1,EGFR,SRC,IGF1R,PPARG.

Establishment of a prediction model for severe acute radiation enteritis associated with cervical cancer radiotherapy

BACKGROUND Cervical cancer is one of the most common gynecological malignant tumors.Radiation enteritis(RE)leads to radiotherapy intolerance or termination of radiotherapy,which negatively impacts the therapeutic effect and seriously affects the quality of life of patients.If the incidence of RE in patients can be predicted in advance,and targeted clinical preventive treatment can be carried out,the side effects of radiotherapy in cervical cancer patients can be significantly reduced.Furthermore,accurate prediction of RE is essential for the selection of individualized radiation dose and the optimization of the radiotherapy plan.AIM To analyze the relationships between severe acute RE(SARE)of cervical cancer radiotherapy and clinical factors and dose-volume parameters retrospectively.METHODS We included 50 cervical cancer patients who received volumetric modulated arc therapy(VMAT)from September 2017 to June 2018 in the Department of Radiotherapy at The First Affiliated Hospital Soochow University.Clinical and dose-volume histogram factors of patients were collected.Logistic regression analysis was used to evaluate the predictive value of each factor for SARE.A nomogram to predict SARE was developed(SARE scoring system≥3 points)based on the multiple regression coefficients;validity was verified by an internal verification method.RESULTS Gastrointestinal and hematological toxicity of cervical cancer VMAT gradually increased with radiotherapy and reached the peak at the end of radiotherapy.The main adverse reactions were diarrhea,abdominal pain,colitis,anal swelling,and blood in the stool.There was no significant difference in the incidence of gastrointestinal toxicity between the radical and postoperative adjuvant radiotherapy groups(P>0.05).There were significant differences in the small intestine V_(20),V_(30),V_(40),and rectal V40 between adjuvant radiotherapy and radical radiotherapy after surgery(P<0.05).Univariate and multivariate analyses revealed anal bulge rating(OR:14.779,95%CI:1.281-170.547,P=0.031)and disease activity index(DAI)score(OR:53.928,95%CI:3.822-760.948,P=0.003)as independent predictors of SARE.CONCLUSION Anal bulge rating(>0.500 grade)and DAI score(>2.165 points)can predict SARE.The nomogram shows potential value in clinical practice.

Effects of tannic acid on the immunity and intestinal health of broiler chickens with necrotic enteritis infection

Background In broiler chickens,necrotic enteritis(NE)infection can reduce production performance.Tannic acid has shown great potential as a treatment of NE in broilers.However,the appropriate dosage of tannic acid in NE of broilers and the improvement effect on intestinal health are not very clear.In this study,we aimed to investigate the effects of different doses of tannic acid on the production performance,immunity,and intestinal health of broilers by constructing an NE model with C.perfringens infection and determining the appropriate dosage of tannic acid with regard to NE.Results Challenged birds showed significant reduction in body weight,villus height,and the ratio of villus height to crypt depth(P<0.05)and increase in the feed consumption gain ratio,intestinal lesion score,and crypt depth(P<0.05).The infection significantly reduced the relative Bacteroidota and Ligilactobacillus abundance(P<0.05)and increased the ratio of Firmicutes/Bacteroidota and cecal content of C.perfringens(P<0.05).Challenged birds fed diets supplemented with tannic acid showed significantly increased mRNA expression of nutrient transport carriers and intestinal barrier genes and growth performance and reduced serum zonulin and endotoxin levels(P<0.05).Addi-tion of tannic acid to the diet inhibited the inflammatory response by reducing the number of coccidia oocysts in feces and the content of C.perfringens in the cecum.Specifically,tannic acid reduced the serum levels of C reactive protein,myeloperoxidase,and specific IgY and ileal mucosal secretory immunoglobulin A levels in the ileal mucosa compared with those in the NE-infected birds.NE-infected birds fed diets supplemented with tannic acid also showed significantly increased relative Anaerocolumna,Thermoanaerobacterium,and Thermosinus abundance(P<0.05);their microbial composition and functional predictions were similar to those of the NC group.Conclusions Tannic acid in the diet alleviated NE by enhancing the intestinal barrier and absorption function.The recommended dietary tannic acid additive level is 500–750 mg/kg.Our study findings would be useful in reducing related economic losses in the broiler industry.

Stimbiotic supplementation modulated intestinal inflammatory response and improved broilers performance in an experimentally-induced necrotic enteritis infection model

Background:Two experiments were conducted to establish an optimal NE challenge model and evaluate the efficacy of stimbiotic(STB)supplementation in necrotic enteritis(NE)challenged broilers.In Exp.1,a total of 120 Arbor Acres(AA)broilers(45.0±0.21 g)were randomly assigned to 6 treatments in a 3×2 factorial arrangement.Vaccine treatments included non-challenge(0),×10 the recommended dose(×10)or×20 the recommended dose(×20)by the manufacturer.Clostridium perfringens(CP)treatments were non-challenge(No)or 3 mL of 2.2×10^(7)CFU CP challenge(Yes).In Exp.2,a total of 72 AA broilers(40.17±0.27 g)were randomly assigned to 6 treatments in a 3×2 factorial arrangement.Dietary treatments included non-additive(CON),100 mg/kg STB(STB)and 100 mg/kg STB on top of a typical commercial blend including an essential oil,probiotics,and enzyme(CB).Challenge treatments included non-NE challenge(No)and NE challenge(Yes)as established in Exp.1.Results:In Exp.1,CP and vaccine challenge decreased(P<0.05)body weight(BW),body weight gain(BWG)and feed intake(FI),and increased(P<0.05)the number of broilers with diarrhea and intestinal lesions.The oral administration of×20 recommended dose of vaccines coupled with 3 mL of 2.2×10^(7)CFU CP resulted in(P<0.01)a significantly increased incidence of wet litter and intestinal lesions.Thus,this treatment was chosen as the challenge model for the successful inducement of NE in Exp.2.In Exp.2,the NE challenge negatively affected(P<0.01)growth performance,ileal morphology,immunoglobulin contents in blood,caecal microbiota in the caecum,footpad dermatitis,intestinal lesion scores,tumour necrosis factor(TNF-α)and endotoxin in the serum compared with the non-NE challenged birds.The supplementation of STB and CB in diets enhanced(P<0.05)growth performance,intestinal microbiota,and blood profiles by stimulating ileal morphology(VH and VH:CD)and propionate production in the cecum,and there were no differences in measured variables between STB and CB supplemented birds.Conclusion:Overall,these results indicate that STB supplementation was able to reduce the inflammatory response and improve the performance of NE challenged birds,and the supplementation of STB alone was as effective as a typical commercial blend containing a number of other additives.

Essential Oils as an Alternative to Antibiotics to Reduce the Incidence and Severity of Necrotic Enteritis in Broiler Chickens: A Short Review

Due to the removal of antibiotic growth promoters (AGPs) and consumer pressure for antibiotic-free (ABF) or no antibiotics ever (NAE) poultry production, there is a need for sustainable alternatives to prevent disease in commercial poultry operations. Without AGPs, there has been a rise in diseases that were traditionally controlled by subtherapeutic levels of antibiotics in the diet. This has impacted the health of commercial poultry and has been a significant cost to poultry producers. To mitigate this, the industry has started to investigate alternatives to antibiotics to treat these forthcoming health issues, such as necrotic enteritis (NE). NE is an enteric disease caused by an over proliferation of toxigenic Clostridium perfringens (CP) in the gastrointestinal tract. Although CP is a commensal in the avian intestinal tract, dysbiosis caused by inflammation and impaired intestinal integrity facilitates uncontrolled replication of CP. Infectious agents, such as Eimeria maxima, appear to be a predominant predisposing factor that promotes NE. However, non-infectious stressors, including dietary changes, have also been associated with NE to some degree. As a result of increased pressure to restrict the use of antibiotics, there is a need for research evaluating the efficacy of alternatives, such as plant-derived essential oils, as potential tools to mitigate NE in commercial poultry flocks. The aim of this study is to review the effects of essential oils as an alternative to antibiotics to reduce the incidence and severity of necrotic enteritis in broiler chickens.

Rare case of lupus enteritis presenting as colorectum involvement:A case report and review of literature

BACKGROUND Systemic lupus erythematosus(SLE)is a multisystem autoimmune disease that can affect the gastrointestinal tract.Most cases of lupus enteritis(LE)involve the small intestine,while the involvement of the whole colon and rectum without the small intestine being affected is extremely rare.CASE SUMMARY A 35-year-old woman was diagnosed with colorectal LE after initially presenting with intermittent abdominal pain and vomiting for two months.She had a regular medication history for five years following the diagnosis of SLE but had been irregular in taking medications,which may have contributed to the onset of LE and led to her current hospital admission.According to the 2019 Classification criteria for SLE of the European League Against Rheumatism/American College of Rheumatology,this case scored 14.Additionally,abdominal computed tomography revealed significant wall edema of the colon and rectum,ischemia and hyperemia of the ascending colon intestinal wall,mesenteric vessel engorgement,increased mesangial fat attenuation,ascites,and bilateral ureter-hydronephrosis,all indicative of colon and rectum LE.Laboratory tests also showed lower levels of complement C3 and C4,with an antinuclear antibody titer of 1:100.Overall,it was clear that this case involved the colon and rectum without affecting the small intestine,representing a rare manifestation of SLE.The patient received treatment with 10 mg of methylprednisolone sodium succinate,100 mL of 0.9%sodium chloride,hydroxychloroquine(100 mg),and nutrition support.After one week of methylprednisolone and hydroxychloroquine therapy,her SLE symptoms and disease activity improved significantly.CONCLUSION Although colorectal LE without small intestine involvement is very rare,early diagnosis and excellent management with corticosteroids prevented the need for surgical intervention.Physicians should be aware of colorectal LE without small intestine involvement as a manifestation of lupus flare.

Adult eosinophilic gastroenteritis and hypereosinophilic syndromes

Eosinophilic gastroenteritis (EGE) in the adult is a distinctive pathologically-based disorder characterized by an eosinophil-predominant mucosal inflammatory process. Most often,the disorder is detected during endoscopic investigation for abdominal pain or diarrhea. Other causes of gastric and intestinal mucosal eosinophilia require exclusion,including parasitic infections and drug-induced causes. Occasionally,the muscle wall or serosal surface may be involved. EGE appears to be more readily recognized,in large part,due to an evolution in the imaging methods used to evaluate abdominal pain and diarrhea,in particular,endoscopic imaging and mucosal biopsies. Def inition of EGE,however,may be diffi cult,as the normal ranges of eosinophil numbers in normal and abnormal gastric and intestinal mucosa are not well standardized. Also,the eosinophilic inflammatory process may be either patchy or diffuse and the detection of the eosinophilic infiltrates may vary depending on the method of biopsy fixation. Treatment has traditionally focused on resolution of symptoms,and,in some instances,eosinophil quantification in pre-treatment and post-treatment biopsies. Future evaluation and treatment of EGE may depend on precise serological biomarkers to aid in defi nition of the long-term natural history of the disorder and its response to pharmacological or biological forms of therapy.

Modification of end-loop ileostomy for the treatment of ischemic or radiation enteritis

AIM: To evaluate a new technique of temporary ileal anastomotic stoma, following small bowel resection, in patients where the anastomosis is anticipated to have borderline margins with dubious viability. METHODS: Five patients underwent enterectomy and partially anastomosed end-loop ileostomy at the University Hospital of Larissa between 2000 and 2006. Enterectomy was performed because of conditions such as mesenteric vascular occlusive disease, radiation entritis and small bowel injury. RESULTS: Postoperatively, none of the patients developed any stoma-related or anastomotic complications. There were no major complications. All patients were discharged between the 8th and 15th day after the procedure, and the stoma was closed 3 wk to 4 wk later. CONCLUSION: We believe that our proposed modification of end-loop ileostomy is a simple, quick and safe technique with minimal stoma-related morbidity, and with simple and safe reversion. This technique can be considered as a useful option in the treatment of ischemic or radiation-induced enteritis, and in the management of severe intestinal trauma.

Enterolithiasis complicating eosinophilic enteritis:A case report and review of literature

We report a case of eosinophilic enteritis involving the proximal small bowel,a relatively rare entity,presenting unusually as enteroliths in a 68-year-old man with complaints of anemia,malena and abdominal pain.The disease if diagnosed in the initial stages responds well to medical treatment but if associated with complications or misdiagnosed,surgical modality is the treatment of choice.In our case,the patient presented with enteroliths and strictures.Resection and anastomosis of the small bowel containing stones was carried out.Histopathology conf irmed the diagnosis as eosinophilic enteritis.

Detection of apoptosis induced by new type gosling viral enteritis virus in vitro through fluorescein annexin V-FITC/PI double labeling

AIM: To achieve a better understanding of the pathogenesis of new type gosling viral enteritis virus (NGVEV) and the relationship between NGVEV and host cells. METHODS: The apoptosis of duck embryo fibroblasts (DEF) induced by NGVEV was investigated by fluorescence-activated cell sorter (FACS) and fluorescence microscope after the cells were stained with Annexin V-FITC and propidium iodide (PI). RESULTS: By staining cells with a combination of fluorescein annexin V-FITC and PI, it is possible to distinguish and quantitatively analyze non-apoptotic cells (Annexin V-FITC negative/PI negative), early apoptotic cells (Annexin V-FITC positive/PI negative), late apoptotic/necrotic cells (Annexin V-FITC positive/ PI positive) and dead cells (Annexin V-FITC negative/PI positive) through flow cytometry and fluorescence microscope. The percentage of apoptotic cells increased with the incubation time and reached a maximum at 120 h after infection, while the percentage of non- apoptotic cells decreased.

Infliximab induces remission in cryptogenic multifocal ulcerous stenosing enteritis:First case

We present the case of a 29-year-old patient with a history of abdominal pain and vomiting.Based on wireless video capsule findings he was previously diagnosed with ileal Crohn's disease at a different institution,although the clinical and radiological picture was not typical and the response to corticosteroids was poor.We performed a single-balloon enteroscopy showing a short,ulcerous stenosis 50 cm proximal from Bauhin's valve.The endoscopic and clinical histopathological findings were compatible with cryptogenic multifocal ulcerous stenosing enteritis(CMUSE).High dose corticosteroids were again started,without effect.The monoclonal tumor necrosis factor-α(TNF-α) antibody infliximab was added to the medical therapy.After induction therapy,both clinical and endoscopic amelioration was obtained.Larger case studies are needed to confirm the efficacy of TNF-α inhibition in steroid refractory CMUSE.

An experimental study of preventing and treating acute radioactive enteritis with human umbilical cord mesenchymal stem cells

Objective:To test the curative effect of human umbilical cord-derived mesenchymal stem cells on rat acute radioactive enteritis and thus in provide clinical therapeutic basis for radiation sickness.Methods:Human umbilical cord-derived mesenchymal stem cells were cultivated in vitro and the model of acute radioactive enteritis of rats was established.Then,the umbilical cord mesenchymal stem cells were injected into the rats via tail vein.Visual and histopathiological changes of the experimental rats were observed.Results:After the injection,the rats in the prevention group and treatment group had remarkably better survival status than those in the control group.The histological observations revealed that the former also had better intestinal mucosa structure,more regenerative cells and stronger proliferation activity than the latter.Conclusions:Human umbilical cord-derived mesenchymal stem cells have a definite therapeutic effect on acute radioactive enteritis in rats.

Role of triamcinolone in radiation enteritis management

AIM:To investigate the role of triamcinolone in the management of acute and chronic enteritis caused by pelvic radiotherapy.METHODS:Twenty-eight patients with rectum adenocarcinoma or endometrium adenocarcinoma were studied.We compared the results of 14 patients treated with injected triamcinolone acetonide(TA)with those of 14 patients who were not treated with TA.For the TA group,40 mg of TA was injected intramuscularly on the 1st,11th and 21st d of radiotherapy;the control group received no injections.All of the study participants had a median age of 65 years,had undergone postoperative radiotherapy and were evaluated weekly using Radiation Therapy Oncology Group and the European Organization for Research and Treatment of Cancer Acute Morbidity Score Criteria,and complete blood counts for every 10 d.RESULTS:Triamcinolone was found to effectively prevent and treat radiation-induced acute gastrointestinal(enteritis)and genitourinary(cystitis)side effects(P=0.022 and P=0.023).For the lower GI side effect follow up,11 patients in the control group had Grade 2toxicity and 3 patients had Grade 1 toxicity.In the TA group,5 patients had Grade 2 toxicity and 9 patients had Grade 1 toxicity.For the genitourinary system side effect follow up,4 patients had Grade 2 toxicity and 6patients had Grade 1 toxicity.Additionally,2 patients had Grade 2 toxicity and 2 patients had Grade 1 toxicity.The neutrophil counts did not differ between the TA group and the control group.There was no meaningful difference between age groups and primary cancers.At the 12th mo of follow up,there were no differences between groups for chronic side effects.CONCLUSION:Triamcinolone is a moderately potent steroid,that is inexpensive and has a good safety profile.It would be beneficial for reducing medical expenses related to treatment of radiation induced enteritis.

Modulations of genes related to gut integrity,apoptosis,and immunity underlie the beneficial effects of Bacillus amyloliquefaciens CECT 5940 in broilers fed diets with different protein levels in a necrotic enteritis challenge model

Background:The ban of in-feed antimicrobial additives has negatively affected the poultry industry by causing necrotic enteritis(NE)to emerge in the flocks.Alternatives such as Bacillus probiotics have shown to be effective on eliminating the negative effects of this disease.Two experiments were conducted to investigate the effect of Bacillus amyloliquefaciens CECT 5940(BA)in broiler chickens under NE challenge and/or fed diets with different protein levels.Methods:In both experiments,480 day-old mix-sexed Ross-308 broilers were arranged in a 2×2 factorial arrangement of treatments.In experiment 1,the factors were NE challenge(yes or no)and probiotic(yes or no).In experiment 2,the factors were dietary crude protein levels(standard or reduced)and probiotic(yes or no)and were used under NE challenge condition.Oral administration of Eimeria oocysts(day 9)followed by inoculation with Clostridium perfringens(day 14 and 15)was used to induce NE challenge.On day 16,two birds from each treatment were gavaged with fluorescein isothiocyanate-dextran(FITC-d)and blood samples were collected for gut integrity evaluation,and jejunal samples were collected for gene expression assay.Results:In experiment 1,BA supplementation decreased caspase-3(CASP3)(P<0.001)and caspase-8(CASP8)(P<0.05)and increased occludin(OCLD)(P<0.05)expression regardless of the challenge.Additionally,BA supplementation downregulated interfron-γ(IFN-γ)expression(P<0.01)and upregulated immunoglobulin-G(IgG)(P<0.01)and immunoglobulin-M(IgM)(P<0.05)only in challenged birds.In experiment 2,the expression of genes encoding mucin-2(MUC2)(P<0.001),tight junction protein-1(TJP1)(P<0.05)and OCLD(P<0.05)were upregulated by the addition of BA in the diet,regardless of the crude protein level.Further,BA supplementation downregulated INF-γ(P<0.01)and upregulated immunoglobulin-A(IgA)(P<0.05),IgM(P<0.05)and IgG(P<0.01)regardless of the crude protein level.Conclusion:These findings suggest that supplementation of BA in broiler diets can improve gut health by modulation of genes related to the mucosal barrier,tight junction,and immunity in broilers challenged by unfavourable conditions such as NE challenge.

A secondary bile acid from microbiota metabolism attenuates ileitis and bile acid reduction in subclinical necrotic enteritis in chickens

Background:Clostridium perfringens-induced chicken necrotic enteritis(NE)is responsible for substantial economic losses worldwide annually.Recently,as a result of antibiotic growth promoter prohibition,the prevalence of NE in chickens has reemerged.This study was aimed to reduce NE through titrating dietary deoxycholic acid(DCA)as an effective antimicrobial alternative.Materials and methods:Day-old broiler chicks were assigned to six groups and fed diets supplemented with 0(basal diet),0.8,1.0 and 1.5 g/kg(on top of basal diet)DCA.The birds were challenged with Eimeria maxima(20,000 oocysts/bird)at d 18 and C.perfringens(109 CFU/bird per day)at d 23,24,and 25 to induce NE.The birds were sacrificed at d 26 when ileal tissue and digesta were collected for analyzing histopathology,mRNA accumulation and C.perfringens colonization by real-time PCR,targeted metabolomics of bile acids,fluorescence in situ hybridization(FISH),or terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay.Results:At the cellular level,birds infected with E.maxima and C.perfringens developed subclinical NE and showed shortening villi,crypt hyperplasia and immune cell infiltration in ileum.Dietary DCA alleviated the NE-induced ileal inflammation in a dose-dependent manner compared to NE control birds.Consistent with the increased histopathological scores,subclinical NE birds suffered body weight gain reduction compared to the uninfected birds,an effect attenuated with increased doses of dietary DCA.At the molecular level,the highest dose of DCA at 1.5 g/kg reduced C.perfringens luminal colonization compared to NE birds using PCR and FISH.Furthermore,the dietary DCA reduced subclinical NE-induced intestinal inflammatory gene expression and cell apoptosis using PCR and TUNEL assays.Upon further examining ileal bile acid pool through targeted metabolomics,subclinical NE reduced the total bile acid level in ileal digesta compared to uninfected birds.Notably,dietary DCA increased total bile acid and DCA levels in a dose-dependent manner compared to NE birds.Conclusion:These results indicate that DCA attenuates NE-induced intestinal inflammation and bile acid reduction and could be an effective antimicrobial alternative against the intestinal disease.

Diaphragm disease compared with cryptogenic multifocal ulcerous stenosing enteritis

As the use of drugs such as nonsteroidal anti-inflammatory drugs (NSAIDs) increases,so too do gastrointestinal ulcers,bleeding,perforation and obstruction.Diaphragm disease of the small intestine is formed by submucosal fibrosis and destruction of lamina muscularis due to chronic ulceration,which corresponds to the most severe stage of NSAID enteropathy.It may lead to stricture of the small intestine.If such ulcerations and strictures in the small intestine are multiple,differential diagnosis is between diaphragm disease and cryptogenic multifocal ulcerous stenosing enteritis (CMUSE),because the gross findings of diaphragm disease are similar to those of CMUSE.We report a rare case of diaphragm disease caused by NSAID.It has been finally confirmed by capsule endoscopy and the origin of chronic obscure gastrointestinal bleeding was found to be multiple ulcers and strictures in the small intestine.After operation,we diagnosed the patient with diaphragm disease rather than CMUSE.

Pathogenic and pathological characteristic of new type gosling viral enteritis first observed in China

AIM: To study the purifying method and characteristics of new gosling viral enteritis virus (NGVEV),the etiological agent of new gosling viral enteritis (NGVE) which was first recognized in China, as well as the pathomorphological development in goslings infected artificially with NGVEV. METHODS: (1)NGVEV virions were purified by the procedure of treatment with chloroform and ammonium sulfate precipitation, dialysis to remove the sulfate radical and ammonium ion and separation by gel filtration chromatography, and SDS-PAGE. (2)Forty-2-day-old White Sichuan goslings were orally administered with NGVEV and 24 hr later 2 birds were randomly selected and killed at 24hr intervals until death occurred. Specimens(duodenum, ileum, liver, heart, kidney, spleen, lung, proventriculus, pancreas, esophagus, and the intestinal embolus) were taken until all birds in this group died and were sectioned and stained with hemotoxylin and eosin and studied by light microscope. RESULTS: NGVEV shared the typical characteristics of Adenovirus and which structural proteins consisted of 15 polypeptides. Necrosis and sloughing of the epithelial cells covering the villus tips of the duodenum were first observed in goslings 2 days postinfection artificially with NGVEV. With the progress of infection, this lesion rapidly occurred in the epithelium at the base of the villus and with infiltration of the inflammatory cells, the jejunum tended to be involved. With the intensification of mucosa necrosis and inflammatory exudation of the small intestine, fibrinonecrotic enteritis was further developed and embolus composed of either intestinal contents wrapped by pseudo-membrane or of the mixture of fibrous exudate and necrotic intestinal mucosa were observed in the middle-lower part of the small intestine. This structure occluded the intestinal tract and made the intestine dilated in appearance. The intestinal glandular cells underwent degeneration, necrosis and might be found sloughed into the lumen. Hemorrhage and hyperemia could be observed on the lung and kidney. Epithelial cells of the renal tubular underwent degeneration. In some cases, granular degeneration and fatty degeneration could be found in the liver and in some cases at a later stage of this disease the epithelial cells of trachea and proventriculus might be found sloughed. In some cases at an early stage of this disease, cardiac hyperemia and hemorrhage could be observed. Esophagus, pancreas and brain were found normal. Analyses and comparisons between the pathologic lesions of NGVE and Gosling Plague (GP) were available in this paper as well. CONCLUSION: (1)NGVEV is adenovirus. (2)Pathological characteristic could be as the data for NGVE diagnosis.