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This work aimed to develop a nanosuspension (NSps) as an acetogenin (ACGs) carrier, using soy lecithin (SL) and hydroxypropyl-β-cyclodextrin (βCD) named NSps-βCDSL-ACGs. It was characterized by various spectroscopic techniques (DLS, FTIR, UV-vis diffuse reflectance). Moreover, the NSps morphology was observed by transmission electron microscopy (TEM). Also, the antibacterial activity of NSps-βCDSL-ACGs was evaluated against strains of interest in the poultry sector. NSps-βCDSL-ACGs presented nanometric size (207-239 nm), acceptable polydispersity index (PDI) values (0.13-0.17) and a high Z potential value (-47.17-50.36 mV), demonstrating high stability. The presence of ACGs in NSps-βCDSL-ACGs was confirmed by FTIR analysis. The nanoparticles had a spherical shape and exhibited high inhibition potential against Salmonella Enteritidis (88.18%), Streptococcus gallolyticus (88.01%), Salmonella Typhimurium (86.28%) and Salmonella Infantis (77.02%) strains up to 48 h, and a reduction of up to 3 log CFU/mL was achieved for S. Typhimurium. Therefore, NSps-βCDSL-ACGs is an attractive option for implementing ACGs administration in the poultry sector to reduce the use of antibiotics and minimize bacterial resistance.

Salmonellae remain a prominent issue for poultry products regarding food safety. A predominant source of broiler carcass contamination at processing is from crop rupturing and subsequent contamination of the carcass during evisceration. Chlorhexidine (CHX) salts are commonly used in dental products for their antimicrobial capabilities. We evaluated CHX gluconate and CHX digluconate alongside cetylpyridinium chloride (CPC), a common disinfectant to test their antimicrobial properties and any possible synergistic effects between the two. These effects were evaluated using an in vitro assay simulating the crop contents of poultry. CHX digluconate, CHX gluconate, commercially available CHX gluconate and CPC were tested at 0.1 %, 0.2 %, 1 %, 1.8 %, and 2 %. Borosilicate tubes containing 2.5 g of chicken mash feed, pasteurized to remove unwanted bacteria, were used for assay. Pasteurized feed within tubes were suspended in CHX, CPC, CHX + CPC, or saline vehicle alone (control). Tubes were then inoculated with Salmonella Typhimurium (ST) (except non-inoculated controls) and allowed to statically incubate for 2 h at 37°C. Tube contents were then serially diluted for CFU enumeration. CHX and CPC, when used individually, demonstrated meaningful and significant (p < 0.05) reduction in ST recovery, especially at higher concentrations (1-2 %). Combinations of CHX + CPC at lower concentrations (0.1-0.2 %) were shown to have major antimicrobial effects and either reduced (p < 0.05) or eliminated ST recovery in this assay. These lower concentrations might be used for antemortem disinfection of crop contents prior to live haul at processing in future studies. In vivo efficacy may be affected by limitations on voluntary ingestion of drinking water during pre-slaughter feed withdrawal and was not evaluated in these studies., Competing Interests: Disclosures The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

This article follows-up on our recently published work, which evaluated the impact of the addition of an alfalfa leaf-derived adsorbent in the aflatoxin B 1 (AFB 1 )-contaminated diet in regard to the production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology of turkey poults. This paper presents complementary results on microbial community, ileal morphology, barrier function, and immunity. For this purpose, 350 1-day-old female turkey poults were randomly distributed into five groups: (1) Control, AFB 1 -free diet; (2) AF, AFB 1 -contaminated diet at 250 ng/g; (3) alfalfa, AFB 1 -free diet + 0.5% ( w / w ) adsorbent; (4) alfalfa + AF, AFB 1 -contaminated diet at 250 ng/g + 0.5% ( w / w ) adsorbent; and (5) YCW + AF, AFB 1 -contaminated diet at 250 ng/g + 0.5% ( w / w ) commercial yeast cell wall-based adsorbent (reference group). In general, in the AF group, the growth of opportunistic pathogens was promoted, which lead to gut dysbacteriosis, mainly influenced by Streptococcus lutetiensis . Conversely, a significant increase in beneficial bacteria ( Faecalibacterium and Coprococcus catus ) was promoted by the addition of the plant-based adsorbent. Moreover, the AF group had the lowest villus height and a compromised barrier function, as evidenced by a significant ( p < 0.05) increase in fluorescein isothiocyanate dextran (FITC-d), but these negative effects were almost reversed by the addition of the alfalfa adsorbent. Furthermore, the AF + YCW and alfalfa + AF groups exhibited a significant increase in the cutaneous basophil hypersensitivity response compared to the rest of the experimental groups. Taken together, these results pointed out that the alfalfa counteracts the adverse effects of AFB 1 in poults, facilitating the colonization of beneficial bacteria and improving the barrier function of the turkey poults.

This experiment was conducted to determine the effect of an adsorbent material based on powdered alfalfa leaves added in the aflatoxin B 1 (AFB 1 )-contaminated diet of turkey poults on production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology. For this purpose, three hundred and fifty female Nicholas-700 poults were randomly assigned into five treatments: (1) Control, AFB 1 -free diet; (2) AF, diet contaminated with 250 ng AFB 1 /g; (3) Alfalfa, AFB 1 -free diet + 0.5% (w/w) adsorbent; (4) AF+alfalfa, diet contaminated with 250 ng AFB 1 /g + 0.5% (w/w) adsorbent, and (5) AF+ yeast cell wall (YCW), diet contaminated with 250 ng AFB 1 /g + 0.5% (w/w) of yeast cell wall (a commercial mycotoxin binder used as reference material). The in vivo efficacy of powdered alfalfa leaves was assessed during a 28-day period. In general, the addition of powdered alfalfa leaves in the AFB 1 -free diet gave the best performance results (body weight, body weight gain, and feed intake) and improved the values of total protein, glucose, calcium, creatinine, and blood urea nitrogen. Moreover, the addition of powdered alfalfa leaves in the AFB 1 -contaminated diet enhanced body weight and body weight gain and significantly reduced the feed intake, compared to the AF and AF+YCW groups. Additionally, significant alterations in serum parameters were observed in poults intoxicated with the AFB 1 , compared to the Control group. Furthermore, typical histopathological lesions were observed in the liver of the AF group, which were significantly ameliorated with the addition of powdered alfalfa leaves. Conclusively, these results pointed out that low inclusion of powdered alfalfa leaves in the contaminated feed counteracted the adverse effects of AFB 1 in turkey poults., (© 2024. The Author(s).)

Birds, including canaries and other birds, have become increasingly popular as pets. Bird fairs, where breeders gather and show their production in a championship setting, present a setting for possible Salmonella spp. contamination and transmission. Therefore, this study estimated the rate of Salmonella spp. isolation from cage papers, located in the bottom of cages of exotic pet birds, including canaries. Collected Salmonella isolates were used to determine the antimicrobial resistance profile to 52 antibiotics and 17 commercial disinfectants, based on pure or a mixture of acids, alcohols, aldehydes, alkalis, halogens, peroxygen, and quaternary ammonium compounds. The samples consisted of 774 cage papers taken in the 2015 Argentinean canary breeder championship, pooling three cage papers into one sterile sampling bag. Only one pool of the cage papers was positive for Salmonella spp. (0.4%), which belonged to the sample from three frill canary cages. Two strains of Salmonella serotype Glostrup were isolated, which were only resistant to sulfonamides and erythromycin and sensitive to alkali-based product PL301 AS. Although the rate of Salmonella spp. isolation from cage papers in an Argentinean canary breeder championship is low, it should not be discounted because Salmonella ser. Glostrup can be a source of human Salmonella outbreaks and they show high resistance to disinfecting products.

A recent study published data on the growth performance, relative weights of the organs of the gastrointestinal tract, liver histology, serum biochemistry, and hematological parameters for turkey poults fed an experimental diet contaminated with aflatoxin B 1 (AFB 1 ) and humic acids (HA) extracted from vermicompost. The negative effects of AFB 1 (250 ng AFB 1 /g of feed) were significantly reduced by HA supplementation (0.25% w / w ), suggesting that HA might be utilized to ameliorate the negative impact of AFB 1 from contaminated diets. The present study shows the results of the remaining variables, as an extension of a previously published work which aimed to evaluate the impact of HA on the intestinal microbiota, gut integrity, ileum morphometry, and cellular immunity of turkey poults fed an AFB 1 -contaminated diet. For this objective, five equal groups of 1-day-old female Nicholas-700 turkey poults were randomly assigned to the following treatments: negative control (basal diet), positive control (basal diet + 250 ng AFB 1 /g), HA (basal diet + 0.25% HA), HA + AFB 1 (basal diet + 0.25% HA + 250 ng AFB 1 /g), and Zeolite (basal diet + 0.25% zeolite + 250 ng AFB 1 /g). In the experiment, seven replicates of ten poults each were used per treatment ( n = 70). In general, HA supplementation with or without the presence of AFB 1 showed a significant increase ( p < 0.05) in the number of beneficial butyric acid producers, ileum villi height, and ileum total area, and a significant reduction in serum levels of fluorescein isothiocyanate-dextran (FITC-d), a marker of intestinal integrity. In contrast, poults fed with AFB 1 showed a significant increase in Proteobacteria and lower numbers of beneficial bacteria, clearly suggesting gut dysbacteriosis. Moreover, poults supplemented with AFB 1 displayed the lowest morphometric parameters and the highest intestinal permeability. Furthermore, poults in the negative and positive control treatments had the lowest cutaneous basophil hypersensitivity response. These findings suggest that HA supplementation enhanced intestinal integrity (shape and permeability), cellular immune response, and healthier gut microbiota composition, even in the presence of dietary exposure to AFB 1 . These results complement those of the previously published study, suggesting that HA may be a viable dietary intervention to improve gut health and immunity in turkey poults during aflatoxicosis.

The selection of components within a formulation or for treatment must stop being arbitrary and must be focused on scientific evidence that supports the inclusion of each one. Therefore, the objective of the present study was to obtain a formulation based on ascorbic acid (AA) and Eudragit FS 30D microparticles containing curcumin-boric acid (CUR-BA) considering interaction studies between the active components carried out via Fourier transform infrared spectrometry (FTIR) and differential scanning calorimetry (DSC) to minimize antagonistic effects, and comprehensively and effectively treat turkey poults infected with Salmonella enteritidis ( S. enteritidis ). The DSC and FTIR studies clearly demonstrated the interactions between AA, BA, and CUR. Consequently, the combination of AA with CUR and/or BA should be avoided, but not CUR and BA. Furthermore, the Eudragit FS 30D microparticles containing CUR-BA (SD CUR-BA MP) showed a limited release of CUR-BA in an acidic medium, but they were released at a pH 6.8-7.0, which reduced the interactions between CUR-BA and AA. Finally, in the S. enteritidis infection model, turkey poults treated with the combination of AA and SD CUR-BA MP presented lower counts of S. enteritidis in cecal tonsils after 10 days of treatment. These results pointed out that the use of an adequate combination of AA and CUR-BA as an integral treatment of S. enteritidis infections could be a viable option to replace the indiscriminate use of antibiotics.

Enterococcus cecorum (EC) has been associated with septicemia and early mortality in broiler chickens. There is limited research investigating the pathogenicity of EC field strains obtained from affected birds. The purpose of this study was to evaluate the effect of in-ovo administration into the amnion with different EC field isolates at d 18 of embryogenesis (DOE18). In Exp 1, 7 EC field isolates alone or in combination (EC1-EC3, EC4-EC5, EC6, and EC7) were selected based on phenotypic characteristics and evaluated at different concentrations (1 × 10 2 , 1 × 10 4 , and 1 × 10 6 CFU/200 µL/embryo) to assess the impact on early performance and macroscopic lesions. Three isolates (n = 3; EC2, EC5, EC7) were selected for additional evaluation based on the significant (P < 0.05) BWG reduction (d 0-21) compared to the negative control (NC) and the presence of macroscopic lesions observed during posting sessions at d 14 and d 21. An additional isolate associated with enterococcal spondylitis was included in Exp 2 (EC11B). Treatment groups for Exp 2 include: 1) NC, 2) EC2, 3) EC5, 4) EC7, and 5) EC11B (n = 90-120/embryos/group). Groups 2 to 5 were challenged at 1 × 10 2 CFU/200 µL/embryo by in-ovo injection into the amnion at DOE18. Chicks were placed in battery cages for the duration of the study (21 d), and pen weights were recorded at d 0, d 7, d 14, and d 21 to calculate average BW and BWG. At d 14 and d 21 posthatch, liver, spleen, free thoracic vertebrae (FTV), and femoral head (FH) were aseptically collected to enumerate Enterococcus spp. using Chromagar Orientation as the selective media. Cecal contents were collected at d 21 to evaluate the effect of EC challenge on the cecal microbiome composition. There was a significant (P < 0.05) reduction in BW at d 21, and BWG from d 14 to 21 and d 0 to 21, for EC7 and EC11B. Enterococcus cecorum was recovered from the FTV of all challenged groups at d 14 and d 21. The most representative lesions were pericarditis, hydropericardium, focal heart necrosis, and FH osteomyelitis. However, lesions were not uniform across challenged groups or ages (d 14 and d 21). Alpha diversity of the cecal contents was markedly lower in EC5 and EC11B compared to all treatment groups suggesting that EC exposure during late embryogenesis affect the cecal microbiome up to 21 d posthatch. Additionally, these results highlight the differences in pathogenicity of EC strains isolated from field cases and suggest that hatchery exposure to EC during late embryogenesis is a potential route of introduction into a flock., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

This study aims to evaluate the efficacy of humic acid (HA) from worm compost as an adsorbent for aflatoxin B 1 (AFB 1 ) in turkey poults. The experiment involved the inclusion of 0.25% (w/w) HA in the diet of turkey poults consuming aflatoxin-contaminated feed (250 ng AFB 1 /g). A total of 350 1-day-old female Nicholas-700 turkey poults were randomly allocated to five equal groups: negative control (basal diet); positive control (basal diet + 250 ng AFB 1 /g; HA (basal diet + 0.25% HA); HA + AFB 1 (basal diet + HA + 250 ng AFB 1 /g); and zeolite + AFB 1 (basal diet + 0.25% zeolite + 250 ng AFB 1 /g). Each group had seven replicates of 10 poults ( n = 70). The impact of HA addition was evaluated in terms of performance parameters, relative organ weights, liver histological lesions, and serum biochemical and hematological constituents. In general, the addition of HA improved body weight (BW), body weight gain (BWG), and feed conversion rate (FCR). Furthermore, HA effectively mitigated the toxic effects caused by AFB 1 in the majority of the analyzed variables. The results indicated that HA effectively counteracted the AFB 1 -induced toxic effects in turkey poults. Based on these findings, it can be concluded that HA is capable of removing AFB 1 from the contaminated diet., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Maguey-González, Nava-Ramírez, Gómez-Rosales, Ángeles, Solís-Cruz, Hernández-Patlán, Merino-Guzmán, Hernandez-Velasco, Hernández-Ramírez, Loeza, Senas-Cuesta, Latorre, Vázquez-Durán, Du, Méndez-Albores, Hargis and Téllez-Isaías.)

Streptococcus gallolyticus (SG) is a Gram-positive cocci found as commensal gut flora in animals and humans. SG has emerged as a cause of disease in young poults between 1 and 3 wk of age. SG is associated with septicemia resulting in acute mortality with no premonitory signs in turkeys. Three SG isolates were obtained from clinical field cases of acute septicemia of commercial turkeys and used in three independent experiments. In Experiment 1, embryos were inoculated 25 d of embryogenesis with varying concentrations of SG1, SG2, or SG3. In Experiment 2, day of hatch, poults were inoculated with varying concentrations using different routes of administration of SG1, SG2, or SG3. In Experiment 3, day of hatch, poults were inoculated with only isolate SG1 using different paths. Poults were randomly selected for necropsy on d 8 and d 15 and sampled to collect spleen, heart, and liver for SG on d 21, the remaining poults were necropsied and cultured. Samples were plated on Columbia nalidixic acid and colistin agar (CNA) (40°C, 18-24 h). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) confirmed suspect colonies. Data were analyzed using the chi-square test of independence, testing all possible combinations to determine significance (P < 0.05). Weight data were subjected to ANOVA using JMP with significance (P < 0.05). No differences were found in BW or BWG on d 0, 8, 15, or 22. Splenomegaly, focal heart necrosis, and pericarditis were observed in all groups in experiments 1 through 3. In Experiment 3, only airsacculitis was observed in a negative control in separate isolation (P > 0.05). On d 21 of Experiment 3, increased (P < 0.05) recovery of SG from spleens were observed in co-housed negative controls, as well as poults challenged by oral gavage (P > 0.05 for d 7 and d 14). These results confirm numerous previous studies indicating that SG subsp. pasteurianus is a primary infectious microorganism that causes septicemia in young poults., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

A mixed Eimeria spp. challenge model was designed to assess the effects of challenge on broiler chicken performance, intestinal integrity, and the gut microbiome for future use to evaluate alternative strategies for controlling coccidiosis in broiler chickens. The experimental design involved broiler chickens divided into two groups: a control group (uninfected) and a positive control group, infected with Eimeria acervulina (EA), Eimeria maxima (EM), and Eimeria tenella (ET). At day-of-hatch, 240 off-sex male broiler chicks were randomized and allocated to one of two treatment groups. The treatment groups included: (1) Non-challenged (NC, n  = 5 replicate pens); and (2) challenged control (PC, n  = 7 replicate pens) with 20 chickens/pen. Pen weights were recorded at d0, d16, d31, d42, and d52 to determine average body weight (BW) and (BWG). Feed intake was measured at d16, d31, d42, and d52 to calculate feed conversion ratio (FCR). Four diet phases included a starter d0-16, grower d16-31, finisher d31-42, and withdrawal d42-52 diet. At d18, chickens were orally challenged with 200 EA, 3,000 EM, and 500 ET sporulated oocysts/chicken. At d24 (6-day post-challenge) and d37 (19-day post-challenge), intestinal lesion scores were recorded. Additionally, at d24, FITC-d was used as a biomarker to evaluate intestinal permeability and ileal tissue sections were collected for histopathology and gene expression of tight junction proteins. Ileal and cecal contents were also collected to assess the impact of challenge on the microbiome. BWG and FCR from d16-31 was significantly ( p  < 0.05) reduced in PC compared to NC. At d24, intestinal lesion scores were markedly higher in the PC compared to the NC. Intestinal permeability was significantly increased in the PC group based on serum FITC-d levels. Cadherin 1 (CDH1), calprotectin (CALPR), and connexin 45 (Cx45) expression was also upregulated in the ileum of the PC group at d24 (6-day post-challenge) while villin 1 (VIL1) was downregulated in the ileum of the PC group. Additionally, Clostridium perfringens (ASV1) was enriched in the cecal content of the PC group. This model could be used to assess the effect of alternative coccidiosis control methods during the post-challenge with EA, EM, and ET., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Graham, Petrone-Garcia, Hernandez-Velasco, Coles, Juarez-Estrada, Latorre, Chai, Shouse, Zhao, Forga, Senas-Cuesta, Laverty, Martin, Trujillo-Peralta, Loeza, Gray, Hargis and Tellez-Isaias.)

Introduction: Coccidiosis caused by the Eimeria spp., an Apicomplexan protozoon, is a major intestinal disease that affects the poultry industry. Although most cases of coccidiosis are subclinical, Eimeria infections impair bird health and decrease overall performance, which can result in compromised welfare and major economic losses. Viable sporulated Eimeria oocysts are required for challenge studies and live coccidiosis vaccines. Potassium dichromate (PDC) is typically used as a preservative for these stocks during storage. Although effective and inexpensive, PDC is also toxic and carcinogenic. Chlorhexidine (CHX) salts may be a possible alternative, as this is a widely used disinfectant with less toxicity and no known carcinogenic associations., Methods: In vitro testing of CHX gluconate and CHX digluconate exhibited comparable oocyst integrity and viability maintenance with equivalent bacteriostatic and bactericidal activity to PDC. Subsequent use of CHX gluconate or digluconate-preserved Eimeria oocysts, cold-stored at 4°C for 5 months, as the inoculum also resulted in similar oocyst shedding and recovery rates when compared to PDC-preserved oocysts., Results and Discussion: These data show that using 0.20% CHX gluconate could be a suitable replacement for PDC. Additionally, autofluorescence was used as a method to evaluate oocyst viability. Administration of artificially aged oocysts exhibiting >99% autofluorescence from each preserved treatment resulted in no oocyst output for CHX salt groups., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Laverty, Beer, Martin, Hernandez-Velasco, Juarez-Estrada, Arango-Cardona, Forga, Coles, Vuong, Latorre, Señas-Cuesta, Loeza, Gray, Barta, Hargis, Tellez-Isaias and Graham.)

Introduction: Drug-sensitive live coccidiosis vaccines have been used to control coccidiosis and renew drug sensitivity in commercial chicken operations. However, only limited species coverage vaccines have been available for commercial turkey producers. This study aimed to assess the effect of an E. meleagrimitis vaccine candidate, with and without amprolium intervention, on performance and oocyst shedding. Additionally, the effect of vaccination, amprolium treatment, and E. meleagrimitis challenge on intestinal integrity and microbiome composition was evaluated., Methods: Experimental groups included: (1) NC (non-vaccinated, non-challenged control); (2) PC (non-vaccinated, challenged control); (3) VX + Amprol ( E. meleagrimitis candidate vaccine + amprolium); and 4) VX ( E. meleagrimitis candidate vaccine). For VX groups, 50% of the direct poults were orally vaccinated at DOH with 50 sporulated E. meleagrimitis oocysts and were comingled with contact or non-vaccinated poults for the duration of the study. From d10-14, VX + Amprol group received amprolium (0.024%) in the drinking water. All groups except NC were orally challenged with 95K E. meleagrimitis sporulated oocysts/mL/poult at d23. At d29, ileal and cecal contents were collected for 16S rRNA gene-based microbiome analysis., Results and Discussion: VX did not affect performance during the pre-challenge period. At d23-29 (post-challenge), VX groups had significantly ( P < 0.05) higher BWG than the PC group. Contacts and directs of VX groups in LS had significantly reduced compared to PC. As anticipated, amprolium treatment markedly reduced fecal and litter OPG for the VX + Amprol group compared to the VX group which did not receive amprolium. The ileal and cecal content results showed that the PC group had different bacterial diversity and structure, including alpha and beta diversity, compared to NC. Linear discriminant analysis Effect Size (LEfSe) identified that Lactobacillus salivarius (ASV2) was enriched in PC's ileal and cecal content. Compared to NC and PC, the vaccinated groups showed no distinct clusters, but there were similarities in the ileal and cecal communities based on Bray-Curtis and Jaccard distances. In conclusion, these results indicate that vaccination with this strain of E. meleagrimitis , with or without amprolium intervention, caused a very mild infection that induced protective immunity and challenge markedly affected both the ileal and cecal microbiome., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Trujillo-Peralta, Latorre, Chai, Senas-Cuesta, Forga, Coles, Zhao, Hernandez-Velasco, Tellez-Isaias, Barta, Bielke, Hargis and Graham.)

Essential oils (EO) affect performance, intestinal integrity, bone mineralization, and meat quality in broiler chickens subjected to cyclic heat stress (HS). Day-of-hatch Cobb 500 male broiler chicks ( n = 475) were randomly divided into four groups. Group 1: No heat stress (Thermoneutral) + control diets with no antibiotics; Group 2: heat stress control + control diets; Group 3: heat stress + control diets supplemented with thymol chemotype (45 ppm) and herbal betaine (150 ppm) formulation EO1; Group 4: heat stress + control diets supplemented with phellandrene (45 ppm) and herbal betaine (150 ppm) formulation EO2. From day 10-42, the heat stress groups were exposed to cyclic HS at 35°C for 12 h (8:00-20:00). BW, BWG, FI, and FCRc were measured at d 0, 10, 28, and 42. Chickens were orally gavaged with FITC-d on days 10 (before heat stress) and 42. Morphometric analysis of duodenum and ileum samples and bone mineralization of tibias were done. Meat quality was assessed on day 43 with ten chickens per pen per treatment. Heat stress reduced BW by day 28 ( p < 0.05) compared to thermoneutral chickens. At the end of the trial, chickens that received both formulations of EO1 and EO2 had significantly higher BW than HS control chickens. A similar trend was observed for BWG. FCRc was impaired by EO2 supplementation. There was a significant increase in total mortality in EO2 compared with EO1 EO1 chickens had lower FITC-d concentrations at day 42 than the HS control. In addition, EO1 treatment is not statistically different if compared to EO2 and thermoneutral. Control HS broilers had significantly lower tibia breaking strength and total ash at day 42 than heat-stressed chickens supplemented with EO1 and EO2. Heat stress affected intestinal morphology more than thermoneutral chickens. EO1 and EO2 improved intestinal morphology in heat-stressed chickens. Woody breast and white striping were more common in thermoneutral chickens than heat stress chickens. In conclusion, the EO-containing diet could improve broiler chicken growth during cyclic heat stress, becoming increasingly relevant in antibiotic-free production in harsh climates., Competing Interests: Authors AU, BM, and JA-I, were employed by the company Promitec. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Señas-Cuesta, Stein, Latorre, Maynard, Hernandez-Velasco, Petrone-Garcia, Greene, Coles, Gray, Laverty, Martin, Loeza, Uribe, Martínez, Angel-Isaza, Graham, Owens, Hargis and Tellez-Isaias.)

Chronic stress is recognized as a secret killer in poultry. It is associated with systemic inflammation due to cytokine release, dysbiosis, and the so-called leaky gut syndrome, which mainly results from oxidative stress reactions that damage the barrier function of the cells lining the gut wall. Poultry, especially the genetically selected broiler breeds, frequently suffer from these chronic stress symptoms when exposed to multiple stressors in their growing environments. Since oxidative stress reactions and inflammatory damages are multi-stage and long-term processes, overshooting immune reactions and their down-stream effects also negatively affect the animal's microbiota, and finally impair its performance and commercial value. Means to counteract oxidative stress in poultry and other animals are, therefore, highly welcome. Many phytogenic substances, including flavonoids and phenolic compounds, are known to exert anti-inflammatory and antioxidant effects. In this review, firstly, the main stressors in poultry, such as heat stress, mycotoxins, dysbiosis and diets that contain oxidized lipids that trigger oxidative stress and inflammation, are discussed, along with the key transcription factors involved in the related signal transduction pathways. Secondly, the most promising phytogenic substances and their current applications to ameliorate oxidative stress and inflammation in poultry are highlighted.

Outbreaks of histomonosis in turkeys are typically initiated by the ingestion of contaminated embryonated eggs of Heterakis gallinarum , potentially present in earthworms and mechanical vectors. Once an outbreak is started, infected turkeys can transmit the disease by horizontal transmission. Factors influencing horizontal transmission of histomonosis are poorly understood. Replication of horizontal transmission in experimental conditions has not been consistent, presenting an obstacle in searching for alternatives to prevent or treat the disease. Two pilot experiments and three validation experiments were conducted in the present study. In pilot experiment 1, one isolate of Histomonas meleagridis (named Buford) was used. Turkeys were fed a low-nutrient density diet corn-soy based (LOW-CS) and raised in floor pens. In pilot experiment 2, another isolate of H. meleagridis was used (named PHL). Turkeys were fed a low-nutrient density diet with the addition of wheat middlings (LOW-WM) and raised in floor pens. In experiment 3, conducted on floor pens, both isolates and diets were used in different groups. In experiment 4, turkeys were raised on battery cages and only the PHL isolate was used. Both diets (LOW-WM and LOW-CS) were used, in addition to a diet surpassing the nutritional needs of young poults (turkey starter, TS). In experiment 5, conducted in battery cages, only the PHL isolate was used, and the LOW-WM and TS diets were in different groups. The horizontal transmission was achieved only with the PHL isolate from all experiments. The transmission rate varied among experimental diets, with the TS diet having the lowest transmission rate in experiments 4 and 5. Variation was observed between experiments and within experimental groups., Competing Interests: EM is employed by Cargill Turkeys LLC. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Barros, Vuong, Latorre, Cuesta, McGill, Rochell, Tellez-Isaias and Hargis.)

The importance of intestinal alkaline phosphatase (IAP) in maintaining gut health and intestinal homeostasis is well established. The objective of this study was to investigate the tolerance of poultry and swine to dietary supplementation of a novel microbial-derived alkaline phosphatase (AP; E.C. 3.1.3.1 produced by Paenibacillus lentus strain CMG3709). Studies were conducted on day-old Ross 308 chicken ( n = 1,000; Study 1) and weaned piglets ( n = 180; Study 2) for a duration of 42 d; and consisted of four treatment groups (TG) based on the concentration of microbial-derived AP supplemented in their diet at 0; 12,000; 20,000; and 200,000 U/kg of feed. Parameters such as animal survival, hematology, coagulation, and biochemical indices were assessed at the end of the study. The effect of microbial AP on nutrient absorption through skin pigmentation and intestinal permeability were also investigated in broilers ( n = 600; Study 3). In poultry (Study 1), there were no statistically significant differences between control and TG for any of the hematological and biochemical parameters, except for a marginal increase ( P < 0.05) in serum phosphorus at the highest dose. This variation was not dose-dependent, was well within the reference range, and was not associated with any clinical correlates. In swine (Study 2), hematological parameters such as leukocyte, basophil, and lymphocyte counts were lower ( P < 0.05) for the two highest doses but were traced back to individual variations within the group. The biochemical indices in piglets showed no significant differences between control and supplemental groups except for glucose ( P = 0.0005), which showed a high effect ( P = 0.008) of the random blood collection order. Nonetheless, glucose was within the normal reference range, and were not related to in-feed supplementation of AP as they had no biological significance. The survival rate in all three studies was over 98%. Dietary supplementation of microbial-derived AP up to 16.7 times the intended use (12,000 U/kg feed) level had no negative effects in both poultry and swine. In-feed supplementation of microbial-derived AP for 28 d improved intestinal pigment absorption ( P < 0.0001) and reduced intestinal paracellular permeability ( P = 0.0001) in broilers (Study 3). Based on these results, it can be concluded that oral supplementation of microbial-derived AP is safe for poultry and swine and effective at improving gut health in poultry., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science.)

The goal of this research was to assess cyclic heat stress on gut permeability, bone mineralization, and meat quality in chickens. Two separate trials were directed. 320 day-of-hatch Cobb 500 male chicks were randomly assigned to four thermoneutral (TN) and four cyclic heat stress (HS) chambers with two pens each, providing eight replicates per treatment in each trial (n = 20 chicks/replicate). Environmental conditions in the TN group were established to simulate commercial production settings. Heat stress chickens were exposed to cyclic HS at 35 °C for 12 h/day from days 7−42. Performance parameters, intestinal permeability, bone parameters, meat quality, and leukocyte proportions were estimated. There was a significant (p < 0.05) reduction in body weight (BW), BW gain, and feed intake, but the feed conversion ratio increased in chickens under cyclic HS. Moreover, HS chickens had a significantly higher gut permeability, monocyte and basophil levels, but less bone mineralization than TN chickens. Nevertheless, the TN group had significant increases in breast yield, woody breast, and white striping in breast fillets compared to HS. These results present an alternative model to our previously published continuous HS model to better reflect commercial conditions to evaluate commercially available nutraceuticals or products with claims of reducing the severity of heat stress.

Repeated serial in vitro passage of Histomonas meleagridis, the etiological agent of histomoniasis (blackhead) of turkeys, was demonstrated to markedly achieve attenuation and reduction of virulence as compared to the original wild-type isolate. Four experiments were performed to evaluate the route (oral vs. intracloacal) and age (day-of-hatch vs. d 14) for administration of attenuated H. meleagridis isolates as vaccine candidates against homologous or heterologous wild-type challenge. Attenuated H. meleagridis were developed from 2 different strains (Buford strain originating in Georgia; PHL2017 strain originating in Northwest Arkansas). Buford P80a (passage 80, assigned as isolate lineage "a" following repeated passage) was selected as the primary vaccine candidate and was evaluated in Experiments 1-3. Experiment 4 evaluated selected candidates of attenuated PHL2017 (P67, P129) and Buford (P80a, P200a, P138b, P198c) strains against Buford wild-type challenge. As has been demonstrated previously, wild-type H. meleagridis cultures administered orally after 1 day of age were not infective in the current studies, but infection with wild-type cultures could be induced orally at day-of-hatch. Infection was effectively achieved via the intracloacal route at day-of-hatch and in older turkeys (d 21, d 28-29, d 35). Intracloacal inoculation of turkeys with the attenuated passaged isolates as vaccine candidates at d 14 was shown to produce significant (P < 0.05) protection from mortality, reduction in body weight gain, as well as reduction in hepatic and cecal lesions in these experiments following challenge with either the homologous wild-type isolate or from a wild-type strain obtained years later from a geographically disparate area of the United States. Inoculation with the attenuated H. meleagridis isolates at day-of-hatch, either orally or cloacally, did not produce significant protection against subsequent wild-type challenge. While offering significant protection with minimal vaccine-related negative effects, the protection from cloacal vaccine administration was neither significantly robust nor encouraging for industry application using the methods evaluated in the present manuscript since mortalities and lesions were not completely reduced which could thereby potentially allow transmission from residual infection and shedding within a flock., Competing Interests: DISCLOSURES The authors have no conflicts of interest to report., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The three Bacillus strains present in Norum™ were initially selected by their excellent to good relative enzyme activity (REA) production score for amylase, protease, lipase, phytase, cellulase, β-glucanase, and xylanase. Further studies confirmed that the three isolates also showed an antibacterial activity, Gram-positive and Gram-negative poultry pathogens. Norum™ (Eco-Bio/Euxxis Bioscience LLC) is a Bacillus spore direct-fed microbial (DFM). The Bacillus isolates were screened and selected based on in vitro enzyme production profiles. Moreover, in chickens fed high non-starch polysaccharides, this DFM demonstrated to reduce digesta viscosity, bacterial translocation, increase performance, bone mineralization, and balance the intestinal microbiota. In the present study, we present the whole-genome sequence of each of the three isolates in Norum™, as well as the synergistic, additive, or antagonistic effects on the enzyme production behavior of the three Bacillus strains and their combinations when grown together vs. when grown individually. The whole-genome sequence identified isolate AM1002 as Bacillus subtilis (isolate 1), isolate AM0938 as Bacillus amyloliquefaciens (isolate 2), and isolate JD17 as Bacillus licheniformis (isolate 3). The three Bacillus isolates used in the present study produce different enzymes (xylanase, cellulase, phytase, lipase, protease, and β-glucanase). However, this production was modified when two or more Bacillus strains were combined, suggesting possible synergistic, antagonistic, or additive interactions. The Bliss analysis suggested ( p < 0.05) that the combination of Bacillus strains 1-2 and 1-2-3 had intermediate effects and predicted that the combination of Bacillus strains 2-3 could have better effects than the combination of all the three Bacillus strains. In summary, the current study demonstrated the need of selecting Bacillus strains based on quantitative enzyme determination and data analysis to assess the impacts of combinations to avoid antagonistic interactions that could limit treatment efficacy. These results suggest that using Bacillus strains 2-3 together could lead to a new generation of DFMs with effects superior to those already examined in Bacillus strains 1-2-3 and, therefore, a potential alternative to growth-promoting antibiotics. More research utilizing poultry models is being considered to confirm and expand the existing findings., Competing Interests: OO is employed by Biodigest S.A.S., and RD is employed by Nutriavícola S.A. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hernandez-Patlan, Solis-Cruz, Latorre, Merino-Guzman, Morales Rodríguez, Ausland, Hernandez-Velasco, Ortiz Holguin, Delgado, Hargis, Singh and Tellez-Isaias.)

The gut microbiota has been designated as a hidden metabolic 'organ' because of its enormous impact on host metabolism, physiology, nutrition, and immune function. The connection between the intestinal microbiota and their respective host animals is dynamic and, in general, mutually beneficial. This complicated interaction is seen as a determinant of health and disease; thus, intestinal dysbiosis is linked with several metabolic diseases. Therefore, tractable strategies targeting the regulation of intestinal microbiota can control several diseases that are closely related to inflammatory and metabolic disorders. As a result, animal health and performance are improved. One of these strategies is related to dietary supplementation with prebiotics, probiotics, and phytogenic substances. These supplements exert their effects indirectly through manipulation of gut microbiota quality and improvement in intestinal epithelial barrier. Several phytogenic substances, such as berberine, resveratrol, curcumin, carvacrol, thymol, isoflavones and hydrolyzed fibers, have been identified as potential supplements that may also act as welcome means to reduce the usage of antibiotics in feedstock, including poultry farming, through manipulation of the gut microbiome. In addition, these compounds may improve the integrity of tight junctions by controlling tight junction-related proteins and inflammatory signaling pathways in the host animals. In this review, we discuss the role of probiotics, prebiotics, and phytogenic substances in optimizing gut function in poultry.

Background: Interest in the use of natural feed additives as an alternative to antimicrobials in the poultry industry has increased in recent years because of the risk of bacterial resistance. One of the most studied groups are polyphenolic compounds, given their advantages over other types of additives and their easy potentiation of effects when complexes are formed with metal ions. Therefore, the objective of the present study was to evaluate the impact of dietary supplementation of copper acetate (CA), curcumin (CR), and their combination (CA-CR) against Salmonella Typhimurium colonization, intestinal permeability, and cecal microbiota composition in broiler chickens through a laboratory Salmonella infection model. S. Typhimurium recovery was determined on day 10 post-challenge by isolating Salmonella in homogenates of the right cecal tonsil (12 chickens per group) on Xylose Lysine Tergitol-4 (XLT-4) with novobiocin and nalidixic acid. Intestinal integrity was indirectly determined by the fluorometric measurement of fluorescein isothiocyanate dextran (FITC-d) in serum samples from blood obtained on d 10 post-S. Typhimurium challenge. Finally, microbiota analysis was performed using the content of the left caecal tonsil of 5 chickens per group by sequencing V4 region of 16S rRNA gene., Results: The results showed that in two independent studies, all experimental treatments were able to significantly reduce the S. Typhimurium colonization in cecal tonsils (CT, P < 0.0001) compared to the positive control (PC) group. However, only CA-CR was the most effective treatment in reducing S. Typhimurium counts in both independent studies. Furthermore, the serum fluorescein isothiocyanate dextran (FITC-d) concentration in chickens treated with CR was significantly lower when compared to PC (P = 0.0084), which is related to a decrease in intestinal permeability and therefore intestinal integrity. The effect of dietary treatments in reducing Salmonella was further supported by the analysis of 16S rRNA gene sequences using Linear discriminant analysis effect size (LEfSe) since Salmonella was significantly enriched in PC group (LDA score > 2.0 and P < 0.05) compared to other groups. In addition, Coprobacillus, Eubacterium, and Clostridium were significantly higher in the PC group compared to other treatment groups. On the contrary, Fecalibacterium and Enterococcus in CR, unknown genus of Erysipelotrichaceae at CA-CR, and unknown genus of Lachnospiraceae at CA were significantly more abundant respectively., Conclusions: CR treatment was the most effective treatment to reduce S. Typhimurium intestinal colonization and maintain better intestinal homeostasis which might be achieved through modulation of cecal microbiota.

Rye is high in nonstarch polysaccharides (NSP), a complex carbohydrate which cannot be digested by poultry as they lack the endogenous enzymes to do so. Exogenous carbohydrases must therefore be supplemented to avoid the antinutritional effects associated with a high NSP diet. The objectives of the present study were to evaluate the effects of a rye-based diet with and without supplementation of a Bacillus direct-fed microbial (DFM) on body weight, bone mineralization, and leaky gut, as well as its role on influencing serum concentrations of peptide YY (PPY) and the ammonia concentration in turkey manure. Two independent trials were conducted. In each experiment, day-of-hatch female turkey poults were neck tagged and randomly assigned to either a control rye-based diet or a rye-based diet supplemented with the DFM (n = 25 birds/group). At 10 days-of-age, poults in both groups were administered with an appropriate dose of fluorescein isothiocyanate-dextran (FITC-d) by oral gavage. One hour later, all poults were euthanized. Blood was collected to evaluate serum FITC-d and PPY concentrations. Furthermore, in Trial 2 only, both tibias were removed for assessment of bone parameters, and turkey manure was collected to evaluate physicochemical analysis. In both trials, poults treated with the DFM showed a significant increase (P < 0.05) in body weight and body weight gain as compared with control nontreated poults. Poults that received the DFM also had a significant reduction in serum levels of PPY and FITC-d when compared with control nontreated poults. In Trial 2, turkeys treated with the DFM had a substantial increase in tibia strength, tibia diameter, total ash, calcium, and phosphorus when compared with control nontreated turkeys. Their manure was also shown to have a significant reduction in the concentration of ammonia. This is the first report of a commercial DFM reducing the concentration of this compound in turkey manure. In summary, the results of the present study confirm that turkeys fed with a rye-based diet have a significant increase in gut permeability, a reduced body weight, and decreased bone mineralization when compared with turkeys fed with the DFM. Turkeys that received the rye-based diet supplemented with the Bacillus-DFM also had a significant reduction in the serum concentration of PPY when compared with control turkeys. This finding suggests a possible prebiotic effect of rye, warranting future studies to test this effect. Further studies to evaluate the microbiota diversity, as well as the concentration of ceca short-chain fatty acids, are also necessary to confirm the reliability of PPY as a potential metabolomic biomarker in poultry., (Copyright © 2020. Published by Elsevier Inc.)

The effects of in ovo administration of a defined lactic acid microbiota (LAM), previously isolated from adult hens, in the cecae microbiota structure and Enterobacteriaceae colonization after exposure to virulent Escherichia coli during the hatching phase of broiler chickens were evaluated. Embryos inoculated with LAM showed a significant ( P < 0.05) reduction of Enterobacteriaceae colonization at day-of-hatch (DOH) and day (d) 7. Furthermore, there was a significant increase in total lactic acid bacteria on DOH, body weight (BW) DOH, BW d7, and d0-d7 BW gain and reduced mortality d0-d7 was observed in the LAM group compared with that in phosphate-buffered saline (PBS) control. The bacterial composition at the family level revealed that the Enterobacteriaceae was numerically reduced, whereas the Ruminococcaceae was significantly increased in the LAM group when compared with that in the PBS control. Moreover, the bacterial genera Proteus and Butyricicoccus and unidentified bacterial genera of family Lachnospiraceae and Erysipelotrichaceae were significantly enriched in the LAM group. In contrast, the Clostridium of the family Peptostreptococcaceae and unidentified genus of family Enterobacteriaceae were significantly abundant in the PBS control group. In summary, in ovo administration of a defined LAM isolated from adult hens did not affect hatchability, improved body weight gain and reduced mortality at d7, induced variations in the cecae microbiota structure and reduced Enterobacteriaceae colonization on a virulent E. coli horizontal infection model in broiler chickens., (Copyright © 2020 Arreguin-Nava, Graham, Adhikari, Agnello, Selby, Hernandez-Velasco, Vuong, Solis-Cruz, Hernandez-Patlan, Latorre, Tellez, Hargis and Tellez-Isaias.)

Deoxycholic acid (DCA) is a naturally occurring secondary bile acid that originates from intestinal bacterial metabolic conversion of cholate, a primary bile acid. Deoxycholic acid was shown to have antihistomonal properties in vitro, leading to our hypothesis that DCA inclusion within the feed might prevent histomoniasis. Selected dietary concentrations of DCA were evaluated for effects on body weight gain (BWG), lesions, and mortality of turkeys challenged with wild-type Histomonas meleagridis (WTH). Treatments consisted of non-challenged control (NC; basal diet), 0.25% DCA diet + challenge, 0.5% DCA diet + challenge, 1% DCA diet + challenge, and a positive-challenged control (PC; basal diet). All groups were fed a basal starter diet until day 7, at which time DCA diets were administered to the respective groups. On day 14, 2 × 10 5 WTH cells/turkey were intracloacally administered. H. meleagridis-related lesions were evaluated on day 13 post-challenge. Pre-challenge day 0 to 14 BWG was higher (P ≤ 0.05) in the 0.25% DCA group than in the 1% DCA group. There were no significant differences in pre-challenge day 0 to 14 BWG between any of the other groups. No significant differences in mortalities from histomoniasis occurred in the DCA groups as compared to the PC group. No H. meleagridis lesions or mortalities were observed at any time in the NC group. Presence of H. meleagridis-related liver lesions was higher (P ≤ 0.05) in the 0.5% DCA group as compared to the PC group. Using the same controls and experimental timeline, an additional group was included to evaluate a biliogenic diet formulated with 20% whole egg powder to encourage endogenous bile acid production. The biliogenic diet had no statistical impact on pre-challenge day 0 to 14 BWG, but did not reduce H. meleagridis-related mortalities or lesions after the challenge. Taken together, these data suggest that DCA inclusion within the feed at these concentrations and under these experimental conditions does not prevent histomoniasis., (Copyright © 2020. Published by Elsevier Inc.)

Histomoniasis, caused by the protozoan parasite Histomonas meleagridis, is a disease to which turkeys are especially susceptible. Currently, no chemoprophylaxis compounds are available to mitigate this disease. Boric acid (BA) exhibits antifungal, antiseptic, and antiviral properties and has been used in the treatment of yeast infections. Based on these characteristics, an experiment was conducted to evaluate whether BA might be an efficacious prophylaxis against challenge with wild-type H. meleagridis (WTH). On day-of-hatch, poults were randomly assigned to either a basal control diet or a BA diet (basal diet + 0.2% BA). Groups consisted of a nonchallenged control (NC; basal diet), 0.2% BA + challenge (BA; 0.2% BA diet), and a positive-challenged control (PC; basal diet). On day 21, challenged groups were intracloacally inoculated with 2 × 10 5 WTH cells/turkey, and lesions were evaluated on day 14 postchallenge. Individual body weights were recorded on day 0, 21, and 35 to calculate the prechallenge and postchallenge body weight gain (BWG). The BA group resulted in lower prechallenge day 0 to day 21 BWG (P = 0.0001) than the NC group. Postchallenge day 21 to day 35, BWG was also lower (P = 0.0503) in the BA group than the PC group. No differences between the BA and PC groups were detected for mortalities associated with histomoniasis. Moreover, liver and cecal lesions were not statistically different between the BA and PC groups. Taken together, these data suggest that BA was not efficacious in the prevention or reduction of histomoniasis disease severity when provided at 0.2% dietary concentration under these experimental conditions., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Coccidiosis has been a pervasive disease within the poultry industry, with test parameters used to measure effectiveness of treatment strategies often being subjective or influenced by non-disease-related activity. Four experiments were completed, which examined several test parameters of coccidiosis, including body weight gain (BWG), lesion scores, and oocysts per gram of feces (OPG). Each experiment included at least 2 parameters for measuring coccidial infection in chickens and turkeys. In experiment 1, an inoculated control was measured against 3 anticoccidial groups, whereas in experiments 2 to 4, noninoculated and inoculated controls were compared via BWG and OPG. Lesion scores were also included in experiments 1, 3, and 4. Experiment 4 resulted in high correlation, via Pearson correlation coefficient, between BWG and OPG (r = -0.69), very high correlation between OPG and lesion score (r = 0.86), and moderate correlation between BWG and lesion score (r = -0.49). Lesion scores proved to be effective in confirming Eimeria infection, although they did not correlate well with BWG or OPG. Each parameter tended to provide more useful information when lined up with the Eimeria life cycle. Incorporation of OPG, with BWG and lesion scores, as test parameters to measure coccidiosis intervention strategies, provides a global description of disease that may not otherwise be observed with the 2 latter measurements alone., (Copyright © 2019. Published by Elsevier Inc.)

Clostridium perfringens causes severe gastrointestinal diseases, which include necrotic enteritis (NE) in chickens, a deadly disease worldwide. We report here the draft genome sequence of Clostridium perfringens strain TAMU, which was used in developing an NE chicken challenge model. This C. perfringens TAMU genome sequence will aid in advancing potential intervention strategies to reduce NE pathogenesis., (Copyright © 2020 Ausland et al.)

An experiment was conducted to quantify the timing and magnitude of the effects of coccidiosis vaccination on the growth performance, apparent ileal digestibility (AID) of nutrients and energy, intestinal morphology, and plasma carotenoids and nitric oxide in broilers. Treatment groups consisted of 3 coccidiosis control methods [unvaccinated, unmedicated (NC), in-feed chemical coccidiostat (PC), and live oocyst vaccination (VAC) at day of hatch] administered to male Cobb broilers reared in floor pens. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were determined at 12, 16, 20, 28, and 36 d. Blood and ileal digesta were collected from birds in 10 replicate pens of each treatment at 12, 16, 20, and 36 d to evaluate plasma carotenoid and nitric oxide concentrations and determine nutrient AID and IDE. Jejunal samples were taken at 12, 20, and 36 d for morphological measurements. Oocyst shedding in VAC birds was confirmed by increased oocyst counts and decreased carotenoid concentrations ( P < 0.05) when compared with PC birds, with no differences ( P > 0.05) in nitric oxide concentrations. At 20 d, BWG and FI were lowest ( P < 0.05) in VAC birds, intermediate in NC birds, and highest in PC birds, with no differences in FCR ( P > 0.05). By 28 and 36 d, FCR was higher ( P < 0.05) for VAC and NC birds but BWG and FI of VAC birds were similar ( P > 0.05) to PC birds. At d 12, IDE and AID of nitrogen and ether extract were lower ( P < 0.05) in VAC birds than PC birds. At d 16, AID of nitrogen was similar ( P > 0.05) between PC and VAC birds, whereas AID of ether extract remained lower in VAC birds than PC birds. No differences in AID of nutrients or IDE were observed ( P > 0.05) between VAC and PC birds at 20 or 36 d. No differences ( P > 0.05) in jejunal morphology were observed at any time point. Overall, VAC elicited a transient reduction in AID and IDE, particularly for lipids, that diminished by d 20., (Copyright © 2020 Gautier, Latorre, Matsler and Rochell.)

This study evaluated the effect of in ovo Bacillus spp. base probiotic (BBP) administration on hatchability, Gram-negative bacteria (GNB) recovery, performance, and microbiota composition in 2 independent trials using a virulent E. coli seeder challenge model. In each trial, one hundred and eighty 18-day-old embryos were allocated into 1 of 2 groups: Control and treated group (inoculated with 107 BBP). On day 19 of embryogenesis, seeder embryos (n = 18) were inoculated with 4.5 × 104E. coli/mL+272 μg/mL tetracycline and segregated into mesh hatching bags. Twelve chicks per group were euthanized at hatch and at day 7 to evaluate the gastrointestinal composition of total GNB or total aerobic pasteurized bacteria. Also, in trial 2, ceca content from five chickens at day 7 were collected to evaluate microbiota composition. Embryos inoculated with BBP showed a significant (P < 0.05) reduction in the total number of GNB at day-of-hatch (DOH) and day 7. Probiotic treatment increased BW at DOH and day 7, and BW gain (days 0 to 7) when compared with Control chickens. Proteobacteria phylum was significantly reduced, while the Firmicutes was significantly increased by the BBP as compared to the Control (P < 0.05). At family level, Enterobacteriaceae was significantly decreased, while the Lachnospiraceae was significantly elevated in the BBP as compared to the Control group (P < 0.05). The genus Oscillospira was significantly enriched in the BBP group, whereas the unidentified genus of family Enterobacteriaceae in the Control group (P < 0.05). The BBP group increased the bacterial species richness, although there was no significant difference between treatments (P > 0.05). Interestingly, beta diversity showed a significant difference in bacterial community structure between Control and BBP groups (P < 0.05). The results of the present study suggest that in ovo administration of a BBP can reduce the severity of virulent E. coli horizontal transmission and infection of broiler chickens during hatch. The reduction in the severity of the transmission and infection by the BPP might be achieved through alterations of microbiota composition and its community structure., (© 2019 Poultry Science Association Inc.)

The aim of the present study was to evaluate the effect of a commercial Bacillus direct-fed microbial (DFM) on aflatoxin B1 toxic effects, performance, and biochemical and immunologic parameters in broiler chickens. Ninety 1-day-old Cobb 500 male broiler chicks were raised in floor pens for a period of 21 days. Chicks were neck-tagged, individually weighed, and randomly allocated to one of three groups: Negative control (basal feed), aflatoxin B1 (basal feed + 2 ppm AFB1), and DFM (basal feed + 2 ppm AFB1 + Bacillus direct-fed microbial). Each group had three replicates of 10 chickens ( n = 30/group). Body weight and body weight gain were calculated weekly, while feed intake and feed conversion ratio were determined when broilers were 21 days old. On day 21, all chickens were bled, gastrointestinal samples were collected, and spleen and bursa of Fabricius were weighed. This study confirmed that 2 ppm of AFB1 causes severe detrimental effects on performance, biochemical parameters, and immunologic parameters, generating hepatic lesions in broiler chickens ( P < 0.05). However, it was also observed that DFM supplementation provided beneficial effects that might help to improve gut barrier function, anti-inflammatory and antioxidant activities, as well as humoral and cellular immunomodulation. The results of the present study suggest that this Bacillus -DFM added at a concentration of 10 6 spores/gram of feed can be used to counteract the negative effects that occur when birds consume diets contaminated with AFB1, showing beneficial effects on performance parameters, relative organ weights, hepatic lesions, immune response, and serum biochemical variables. The addition of this Bacillus -DFM might mitigate and decrease aflatoxicosis problems in the poultry industry, improving food security, alleviating public health problems, and providing economic benefits. Future studies are needed to fully elucidate the specific mechanisms by which this Bacillus -DFM counteracts the toxic effects of aflatoxin B1.

Two experimental models were conducted to evaluate and compare the effect of ascorbic acid (AA) or curcumin formulated in a solid dispersion (SD-CUR) as prophylactic or therapeutic alternatives to prevent or control S. Enteritidis (SE) infection in broiler chickens. In the prophylactic model, dietary administration of AA showed a significant reduction in SE counts in crop compared to the positive control (PC) group ( p < 0.05), whereas in cecal tonsils (CT), SD-CUR significantly reduced SE recovery. Superoxide dismutase (SOD) activity was significantly higher in chickens supplemented with AA or SD-CUR, and total intestinal IgA levels were significantly lower in both treatments when compared to the PC group. Serum fluorescein isothiocyanate-dextran (FITC-d) levels were reduced by SD-CUR compared to PC, while AA presented significantly lower total aerobic bacteria. In the therapeutic model, only the dietary administration of AA significantly decreased SE in crop and CT on days 3 and 10 post-challenge. FITC-d levels were significantly lower in both treated groups in comparison to PC, but IgA levels were significantly reduced only by AA. The results suggest that dietary AA and SD-CUR have different modes of action to reduce SE intestinal colonization in two different challenge models in broiler chickens.

During the hatching process, chicks are exposed to opportunistic and/or pathogenic organisms, such as virulent or avirulent Escherichia coli. Virulent E. coli strains have not been feasible for induction of neonatal colibacillosis via in ovo challenge due to high embryonic mortality. In this manuscript, we describe the addition and co-administration of the bacteriostatic antibiotic tetracycline to a virulent E. coli challenge culture, improving hatchability and livability of seeder chicks while allowing robust horizontal transmission in the hatching cabinet to contact chicks. Experiment 1 consisted of 3 trials. Experiment 1, trial 1 was conducted to determine an effective ratio of E. coli challenge and tetracycline dose to be utilized in the seeder model. Trials 2 and 3 were conducted to evaluate the transmission of E. coli from seeder to contact chicks. Experiment 2 consisted of 3 independent 7-D trials where body weight gain (BWG), mortality, and selected enteric bacterial recovery were evaluated. In trials 1 to 3, significantly (P < 0.05) more Gram-negative bacteria were recovered from whole gut samples (GIT) vs. negative controls on day of hatch, from both seeder and contact chicks. At day 7 in trial 1, contact chicks had significantly (P < 0.05) more Gram-negative bacteria recovered from the GIT than the negative control, but not in trials 2 and 3. Presumptive lactic acid bacterial recovery was elevated in contact and seeder chicks compared to the negative control in all 3 trials. Contact challenge caused a significant (P < 0.05) reduction in BWG in 2 out of 3 trials at day 7, and there was a significant (P < 0.05) increase in mortality as compared to the negative controls in all trials. These data suggest that co-administration of a virulent E. coli strain with tetracycline allows for hatch of direct challenged chicks and effective horizontal transmission to contact chicks during the hatching process, as evidenced by reduced day 7 performance and altered selected enteric bacterial recovery., (© 2019 Poultry Science Association Inc.)

Necrotic enteritis (NE) caused by Clostridium perfringens infection has reemerged as a prevalent poultry disease worldwide due to reduced usage of prophylactic antibiotics under consumer preferences and regulatory pressures. The lack of alternative antimicrobial strategies to control this disease is mainly due to limited insight into the relationship between NE pathogenesis, microbiome, and host responses. Here we showed that the microbial metabolic byproduct of secondary bile acid deoxycholic acid (DCA), at as low as 50 µM, inhibited 82.8% of C. perfringens growth in Tryptic Soy Broth (P < 0.05). Sequential Eimeria maxima and C. perfringens challenges significantly induced NE, severe intestinal inflammation, and body weight (BW) loss in broiler chickens. These negative effects were diminished (P < 0.05) by 1.5 g/kg DCA diet. At the cellular level, DCA alleviated NE-associated ileal epithelial death and significantly reduced lamina propria cell apoptosis. Interestingly, DCA reduced C. perfringens invasion into ileum (P < 0.05) without altering the bacterial ileal luminal colonization. Molecular analysis showed that DCA significantly reduced inflammatory mediators of Infγ, Litaf, Il1β, and Mmp9 mRNA accumulation in ileal tissue. Mechanism studies revealed that C. perfringens induced (P < 0.05) elevated expression of inflammatory mediators of Infγ, Litaf, and Ptgs2 (Cyclooxygenases-2 (COX-2) gene) in chicken splenocytes. Inhibiting the COX signaling by aspirin significantly attenuated INFγ-induced inflammatory response in the splenocytes. Consistent with the in vitro assay, chickens fed 0.12 g/kg aspirin diet protected the birds against NE-induced BW loss, ileal inflammation, and intestinal cell apoptosis. In conclusion, microbial metabolic product DCA prevents NE-induced BW loss and ileal inflammation through attenuating inflammatory response. These novel findings of microbiome protecting birds against NE provide new options on developing next generation antimicrobial alternatives against NE.

The effect of Lactobacillus spp.-based probiotic candidates on Salmonella enterica serovar Enteritidis (SE) colonization was evaluated in two separate experiments. In each experiment, sixty-one day-of-hatch female turkey poults were obtained from a local hatchery. In both experiments, poults were challenged via oral gavage with 10 4 cfu/poult of SE and randomly allocated to one of two groups ( n = 30 poults): (1) the positive control group and (2) the probiotic treated group. Heated brooder batteries were used for housing each group separately and poults were allowed ad libitum access to water and unmedicated turkey starter feed. 1 h following the SE challenge, poults were treated with 10 6 cfu/poult of probiotic culture via oral gavage or phosphate-buffered saline （PBS）to control groups. A total of 24 h post-treatment, poults were euthanized and the ceca and cecal tonsils from twenty poults were collected aseptically for SE recovery. In both trials, a significant reduction in the incidence and log 10 cfu/g of SE were observed in poults treated with the probiotic when compared with control poults ( p ≤ 0.05). The results of the present study suggest that the administration of this lactic acid-producing bacteria (LAB)-based probiotic 1 h after an SE challenge can be useful in reducing the cecal colonization of this pathogen in neonatal poults., Competing Interests: M. Arreguin-Nava is employed by Eco-Bio LLC. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Restrictions of in-feed antibiotics use in poultry has pushed research toward finding appropriate alternatives such as Direct-Fed Microbials (DFM). In this study, previously tested Bacillus isolates ( B . subtilis and B . amyloliquefaciens ) were used to evaluate their therapeutic and prophylactic effects against Salmonella enterica serovar Enteritidis ( S . Enteritidis) in broiler chickens. For this purpose, initial antibacterial activity of Bacillus -DFM (10 4 spores/g or 10 6 spores/g) against S . Enteritidis colonization in crop, proventriculus and intestine was investigated using an in vitro digestive model. Furthermore, to evaluate therapeutic and prophylactic effects of Bacillus -DFM (10 4 spores/g) against S . Enteritidis colonization, altogether 60 ( n = 30/group) and 30 ( n = 15/group) 1-day-old broiler chickens were randomly allocated to either DFM or control group (without Bacillus -DFM), respectively. Chickens were orally gavaged with 10 4 cfu of S . Enteritidis per chicken at 1-day old, and cecal tonsils (CT) and crop were collected 3 and 10 days later during the therapeutic study, whereas they were orally gavaged with 10 7 cfu of S . Enteritidis per chicken at 6-day-old, and CT and crop were collected 24 h later from two independent trials during the prophylactic study. Serum superoxide dismutase (SOD), FITC-d and intestinal IgA levels were reported for both chicken studies, in addition cecal microbiota analysis was performed during the therapeutic study. DFM significantly reduced S . Enteritidis concentration in the intestine compartment, and in both proventriculus and intestine compartments as compared to the control when used at 10 4 spores/g and 10 6 spores/g, respectively ( p < 0.05). DFM significantly reduced FITC-d and IgA as well as SOD and IgA levels ( p < 0.05) compared to the control in therapeutic and prophylactic studies, respectively. Interestingly, in the therapeutic study, there were significant differences in bacterial community structure and predicted metabolic pathways between DFM and control. Likewise, phylum Actinobacteria and the genera Bifidobacterium, Roseburia, Proteus , and cc&#95;115 were decreased, while the genus Streptococcus was enriched significantly in the DFM group as compared to the control (MetagenomeSeq, p < 0.05). Thus, the overall results suggest that the Bacillus -DFM can reduce S . Enteritidis colonization and improve the intestinal health in chickens through mechanism(s) that might involve the modulation of gut microbiota and their metabolic pathways.

In humans, alterations in bone metabolism have been associated with myopathies. We postulate the hypothesis that perhaps similar pathologies can also be associated in modern chickens. Hence, this study aimed to assess the fat infiltration in bone marrow and its repercussion on broiler chicken affected by Wooden Breast (WB) myopathy. Ten Cobb 500 live birds with extreme rigidity of the Pectoralis major (PM) muscle were selected as WB affected chickens by physical examination of the muscle at 49 days of age, whereas ten chickens healthy with no physical signs of hardness in the breast muscle were considered to be unaffected. Macroscopic lesions in affected chickens included areas of firm and inflamed muscle with pale appearance, hemorrhaging, and viscous exudate on the surface. Bone marrow and sections of the PM muscle were collected and analyzed for light microscopy. Additionally, transmission electron microscopy was conducted in affected or unaffected muscle. Chickens affected with WB showed significant reductions ( P < 0.05) in femur diameter, calcium, and phosphorous percentage but increased breast weight, compression force and filet thickness when compared with non-affected chickens. Interestingly, bone marrow from WB chicken had subjectively, more abundant infiltration of adipose tissue, when compared with non-affected chickens. Histology of the Pectoralis major of birds with WB showed abundant infiltration of adipose tissue, muscle fibers degeneration with necrosis and infiltration of heterophils and mononuclear cells, connective tissue proliferation, and vasculitis. Ultrastructural changes of WB muscle revealed lack definition of bands in muscle tissue, or any normal ultrastructural anatomy such as myofibrils. The endomysium components were necrotic, and in some areas, the endomysium was notable only as a string of necrotic tissue between degraded myofibrils. The fascia appeared hypertrophied, with large areas of necrosis and myofiber without structural identity with degraded mitochondria adjacent to the disrupted muscle tissue. As far as we know, this is the first study that describes a subjective increase in adipose tissue in the bone marrow of chickens affected with WB when compared with non-affected chickens, and reduced bone mineralization.

The objective of this study was to evaluate the of intestinal permeability and liver bacterial translocation (BT) across a modern commercial broiler, a commercial broiler of 1995 genetics, and an unselected Jungle Fowl line. Modern 2015 (MB2015) broiler chicken, random bred line initiated from 1995 (RB1995), and the Giant Jungle fowl (JF). Chickens were randomly allocated to four different dietary treatments. Dietary treatments were (1) a control corn-based diet throughout the trial [corn-corn (C-C)]; (2) an early phase malnutrition diet where chicks received a rye-based diet for 10 days, and then switched to the control diet [rye-corn (R-C)]; (3) a malnutrition rye-diet that was fed throughout the trial [rye-rye (R-R)]; and (4) a late phase malnutrition diet where chicks received the control diet for 10 days, and then switched to the rye diet for the last phase [corn-rye (C-R)]. Paracellular permeability was evaluated using fluorescein isothiocyanate dextran (FITC-D). Liver BT was also evaluated. MB2015 and RB1995 consuming the rye-based diet showed increase serum levels of FITC-D when compared to the corn-fed chickens ( P < 0.05). Overall, MB2015 appeared to have higher enteric permeability than the JF. To our knowledge, this would be the first paper to evaluate the effect of compensatory growth on intestinal permeability and liver BT. Further studies to evaluate microbiome and inflammatory markers in these chicken models are currently being evaluated.

Intestinal health is essential for feed efficiency and growth in animal agriculture and is dependent on barrier function, inflammation and dysbiosis. Our laboratory has published a nutritional model to induce gut inflammation using rye as a source of energy in poultry. More recently, we have used this model as an assessment of a nutritional rehabilitation model for better understanding of childhood undernutrition. The objective of this brief research report was to use a well-establish malabsorption model in broiler chickens using corn and rye as an energy source to identify several intestinal health biomarkers in the serum. To screen for inflammatory biomarkers, seven commercially available tests were used including Griess, superoxide dismutase, thiobarbituric acid reactive substances, Total antioxidant capacity, extracellular-signal-regulated kinase, Citrulline, and Interferon-ɤ; total IgA from cloacal swab was also measured. In the present study, chickens fed rye had a significant ( P < 0.05) reduction in body weight and body weight gain at 10 day when compared with chickens that received the corn diet. In the second phase of the experiment, chickens that remain with the corn diet had significant differences in body weight and body weight gain. No significant differences were observed for any of the four antioxidant biomarkers evaluated in the sera ( P > 0.05). However, significant differences were observed in serum citrulline and IFN-ɤ, as well as in cloacal IgA, in broiler chickens fed with rye, suggesting their potential use as biomarkers to study intestinal inflammation.

Decreases in the use of antibiotics and anticoccidials in the poultry industry have risen the appearance of necrotic enteritis (NE). The purpose of this study was to evaluate the effect of a Bacillus direct-fed microbial (DFM) on growth performance, intestinal integrity, NE lesions and ileal microbiota using a previously established NE-challenged model. At day-of-hatch, chicks were randomly assigned to three different groups: Negative control (NC), Positive control (PC) challenged with Salmonella Typhimurium (day 1), Eimeria maxima (EM, day 13) and Clostridium perfringens (CP, day 18-19), and Bacillus -DFM group (DFM) challenged as the PC. Body weight (BW) and body weight gain (BWG) were measured weekly. Total feed intake (FI) and feed conversion ratio (FCR) were evaluated at day 21. Liver samples were collected to assess bacterial translocation and blood samples were used to measure superoxide dismutase (SOD) and fluorescein isothiocyanate-dextran (FITC-d). Intestinal contents were obtained for determination of total IgA and microbiota analysis. NE lesion scores (LS) were performed at day 21. Chickens consuming the DFM significantly improved BW and had a numerically more efficient FCR compared to PC at day 21. Additionally, there were no significant differences in FCR between the DFM group and NC. Furthermore, the DFM group showed significant reductions in LS, IgA and FITC-d levels compared to the PC. However, there were no significant differences in SOD between the groups. The microbiota analysis indicated that the phylum Proteobacteria was significantly reduced in the DFM group in comparison to PC. At the genus level, Clostridium, Turicibacter, Enterococcus , and Streptococcus were reduced, whereas, Lactobacillus and Bacillus were increased in the DFM group as compared to PC ( p < 0.05). Likewise, the DFM significantly reduced CP as compared to PC. In contrary, no significant differences were observed in bacterial composition between NC vs. DFM. In addition, beta diversity showed significant differences in the microbial community structure between NC vs. PC, and PC vs. DFM. These results suggest that the dietary inclusion of a selected DFM could mitigate the complex negative impacts caused by NE possibly through mechanism(s) that might involve modulation of the gut microbiota.

Two experiments were conducted to evaluate the effect of the prophylactic or therapeutic administration of a 0.1% mixture containing ascorbic acid and a solid dispersion of curcumin with polyvinylpyrrolidone and boric acid (AA-CUR/PVP-BA) against Salmonella Enteritidis ( S . Enteritidis) in broiler chickens. A third experiment was conducted to evaluate the impact of the dietary administration of 0.1% AA-CUR/PVP-BA in a necrotic enteritis (NE) model in broiler chickens. The prophylactic administration of 0.1% AA-CUR/PVP-BA significantly decreased S . Enteritidis colonization in cecal tonsils (CT) when compared to the positive control group (PC, p < 0.05). The therapeutic administration of 0.1% AA-CUR/PVP-BA significantly reduced the concentration of S . Enteritidis by 2.05 and 2.71 log in crop and CT, respectively, when compared with the PC on day 10 post- S . Enteritidis challenge. Furthermore, the serum FITC-d concentration and total intestinal IgA levels were also significantly lower in chickens that received 0.1% AA-CUR/PVP-BA. Contrary, the PC group showed significantly higher total intestinal IgA levels compared to the negative control or AA-CUR/PVP-BA groups in the NE model. However, 0.1% AA-CUR/PVP-BA showed a better effect in reducing the concentration of S . Enteritidis when compared to the NE model. Further studies with higher concentration of AA-CUR/PVP-BA into the feed to extend these preliminary results are currently being evaluated.

The objectives of the present study were to evaluate the antimicrobial effect of boric acid (BA) on Salmonella enteritidis colonization, intestinal permeability, total intestinal IgA levels, and cecal microbiota composition in broiler chickens. For this purpose, sixty day-old-chicks were randomly allocated to one of two groups: 1) non-treated control group and 2) 0.1% (w/w) BA in feed. All chicks were challenged with 10 4  cfu of S. Enteritidis per bird at 1-d old. On day 3 and 10 post-challenge, samples of crop and ceca-cecal tonsils (CCT) were aseptically collected to evaluate overall aerobic and lactic acid bacteria, as well as S. Enteritidis recovery. Blood and intestinal samples were also collected to evaluate intestinal permeability and total intestinal IgA production, respectively. On day 10 post-challenge, the concentration of S. Enteritidis in crop and CCT significantly decreased (P < .05) by 2.60 log10 and 5.26 log10, respectively, in the BA group compared to the control group. Furthermore, chickens treated with 0.1% BA had a significant reduction in serum fluorescein isothiocyanate dextran (FITC-d) concentration and lower total intestinal IgA levels when compared to the control group. Interestingly, in the microbiota analysis, only the abundance of Actinobacteria phylum was significantly lower in the BA group in comparison with the control group and significant differences in beta diversity were also observed between comparing groups. The results of the present study suggest that BA could maintain intestinal homeostasis, as well as the balance in the microbiota due to its effectiveness in controlling S. Enteritidis infection., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

To evaluate the effect of cellulosic polymers (CEL) and curcumin (CUR) on aflatoxin B1 (AFB1) toxic effects on performance, and the biochemical and immunological parameters in broiler chickens, 150 one-day-old male broiler chicks were randomly allocated into five groups with three replicates of 10 chickens per pen: Negative Control (feed); AFB1 (feed + 2 ppm AFB1); CUR (feed + 2 ppm AFB1 + Curcumin 0.2%); CEL (feed + 2 ppm AFB1 + 0.3% Cellulosic polymers); and, CEL + CUR (feed + 2 ppm AFB1 + 0.3% Cellulose polymers + 0.2% Curcumin). Every week, body weight, body weight gain, feed intake, and feed conversion ratio were calculated. On day 21, liver, spleen, bursa of Fabricius, and intestine from five broilers per replicate per group were removed to obtain relative organ weight. Histopathological changes in liver, several biochemical biomarkers, antibody titers, and muscle and skin pigmentation were also recorded. Dietary addition of 0.3% CEL and 0.2% CUR separately significantly diminished some of the toxic effects resulting from AFB1 on performance parameters, relative organs weight, histopathology, immune response, and serum biochemical variables ( P < 0.05); however, the combination of CUR and CEL showed a better-integrated approach for the management of poultry health problems that are related with the consumption of AFB1, since they have different mechanisms of action with different positive effects on the responses of broiler chickens.

Necrotic enteritis (NE) is a recognized multifactorial disease that cost annually to the poultry industry around $2 billion. However, diverse aspects related to its presentation are not completely understood, requiring further studies using known induction experimental models. Therefore, the purpose of this study was to measure the changes occurring in performance, intestinal integrity and ileal microbiome using a previously established NE-challenge model. Chickens were assigned to a negative control group (NC) or a positive control group (PC). In the PC, broilers were orally gavaged with Salmonella Typhimurium (ST) (1 × 10 7 cfu/chick) at day 1, Eimeria maxima (EM) (2.5 × 10 4 oocyst/chick) at day 18 and Clostridium perfringens (CP) (1 × 10 8 cfu/chick/day) at 23-24 days of age. Weekly, body weight (BW), body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) were evaluated. Morbidity and mortality were determined throughout the study, and NE lesion scores were recorded at day 25. Additionally, blood and liver samples were collected to measure gut permeability as determined by levels of serum fluorescein isothiocyanate-dextran (FITC-d) and bacterial translocation (BT). Ileal contents were processed for 16S rRNA gene-based microbiome analysis. Performance parameters and intestinal permeability measurements were negatively impacted in the PC resulting in elevated serum FITC-d and BT with a -6.4% difference in BWG. The NE lesion score in PC (1.97 vs. 0.00) was significantly higher in comparison to NC, although there was no difference in mortality. The microbiome analysis showed a dramatic shift of ileal microbiomes in PC groups as compared to NC (ANOSIM: R = 0.76, P = 0.001). The shift was characterized by reduced abundance of the phylum Actinobacteria ( P < 0.01), and increased abundance of the genera Butyrivibrio, Lactobacillus, Prevotella and Ruminococcus in PC compared to NC ( P < 0.05). Expectedly, Clostridium was found higher in PC (2.98 ± 0.71%) as compared to NC (1.84 ± 0.36%), yet the difference was not significant. In conclusion, results of the present study showed the different intestinal epithelial and microbiological alterations occurring in an established NE-challenge model that considers paratyphoid Salmonella infections in young chicks as an important predisposing factor for presentation of NE.

Aging, poor oxygenation, or improper storage temperature can lead to variable Eimeria oocyst viability, which is not readily assessed microscopically. Under fluorescent microscopy, some aged Eimeria maxima (EM) oocysts were strongly autofluorescent (AF) within the oocystoplasm and sporocysts, whereas others were distinctly non-fluorescent, leading to the hypothesis that non-viable oocysts may be detectible using this simple approach. Using accelerated aging conditions at 45°C, two experiments were conducted to evaluate variable percentages of autofluorescent EM oocysts on body weight gain (BWG), lesion scores (LS), and total oocyst shedding (OS) per bird. Through oral gavage, EM oocysts were administered on d10, whereas LS and BWG were determined d5 post-inoculation. Experiment 1 groups consisted of non-challenged controls (n = 15), or 25,000 EM exhibiting 12.8% (n = 14), 61.1% (n = 10), or 93.3% (n = 10) autofluorescence. BWG in 12.8% AF group was lower (P = 0.054) than non-challenged control. LS were higher (P < 0.05) in 61.1% and 12.8% AF groups as compared to non-challenged control and 93.3% AF groups. Experiment 2 groups consisted of non-challenged controls (LS n = 20/BWG n = 40), or 22,500 EM exhibiting 7% AF (LS n = 20/BWG n = 40), 80.6% AF (LS n = 19/BWG n = 39), or 99% AF (LS n = 19/BWG n = 39). BWG in 7% AF group was lower (P < 0.05) than non-challenged control and 99% AF groups. LS were higher (P < 0.05) in 80.6% and 7% AF groups as compared to non-challenged control and 99% AF groups. OS from d5-8 post-inoculation was determined for each of five replicates per group (n = 20/group; n = 4/replicate), with higher (P < 0.05) OS in 80.6% and 7% AF groups than in non-challenged control or 99% AF groups. Taken together, data indicate lower LS, higher BWG, and reduced OS with higher %AF oocysts, consistent with the hypothesis of lowered viable challenge with this EM isolate.

The purpose of this study was to evaluate the effect of humic acids (HA) on intestinal viscosity, leaky gut and ammonia excretion in a 24 hr feed restriction (FR) model to induce intestinal permeability in chickens. One-day-old male Cobb-Vantress broilers were randomly allocated to one of two groups (n = 25 chickens), with or without 0.2% of isolated HA from worm-compost, and placed in brooder batteries. Chicks had ad libitum access to water and feed for 14 days. Intestinal permeability was induced by 24 hr FR starting at 14 days. At 15 days of age, chickens in both groups were given an appropriate dose of fluorescein isothiocyanate dextran (FITC-d) by oral gavage. Intestine and liver samples were also collected to evaluate viscosity and bacterial translocation (BT), respectively. An increase (p < .05) in intestinal viscosity was observed in the experimental group consuming 0.2% of HA and was confirmed in a published in vitro digestion model that simulates the chemical and physical conditions of the crop, proventriculus and intestine of chickens. Furthermore, the treated group also showed a significant reduction in FITC-d, liver BT and ammonia in the manure. These results suggest that HA have a positive impact in intestinal integrity in chickens., (© 2018 Japanese Society of Animal Science.)