Identification and characterization of the causative agents of Focal Ulcerative Dermatitis in commercial laying hens.

Focal Ulcerative Dermatitis (FUDS) is an emerging dermatological disease that affects cage-free laying flocks, it is characterized by the development of a lesion on the dorsum of the birds; FUDS is sporadic in nature and can result in a drop in egg production and up to 50% of cumulative mortality. A total of two cage-free flocks (flock 1: no history of FUDS; flock 2: birds affected with FUDS) from a commercial laying hen operation in the mid-west U.S. were sampled in this study. The microbial composition of skin, cloacal, cecal, and ileal samples from each bird was characterized through next generation sequencing (NGS). Results identified Staphylococcus aureus and Staphylococcus agnetis as the potential causative agents of FUDS, being the most predominant in FUDS positive birds. These results were confirmed by plating, with both staphylococci as the only pathogens isolated from lesions of FUDS positive birds. A total of 68 confirmed Staphylococcus isolates from skin and environmental samples were further analyzed by whole genome sequencing (WGS) for the presence of antimicrobial resistance (AMR) genes and virulence factors that could have contributed to the development of FUDS. Forty-four-point one-two percent of the isolates had between one and four acquired AMR genes encoding for macrolides, lincosamides, spectrogramines, and beta-lactams resistance. Six classes of virulence factors associated with adherence, enzyme, immune evasion, secretion system, toxin, and iron uptake were identified. The antimicrobial effect of 4 proprietary Bacillus Direct Fed Microbial (DFM) combinations was evaluated against the Staphylococcus aureus and Staphylococcus agnetis isolates, by agar well-diffusion (AWD) assay and competitive exclusion (CE) on broth culture. Through this antimicrobial screening, a particular two-strain combination of Bacillus pumilus was identified as the most effective inhibitor of both staphylococci. A customized Bacillus pumilus product is being used at different farms with history of FUDS resulting in the successful inhibition of both Staphylococcus aureus and Staphylococcus agnetis, decreasing FUDS mortalities, and improving harvestable eggs.

Managing broilers gut health with antibiotic-free diets during subclinical necrotic enteritis.

Necrotic enteritis (NE) caused by Clostridium perfringens is among the most important enteric diseases in poultry production. This study examined the effects of 2 probiotics (Prob) and a synbiotic (Synb) during a naturally occurring NE challenge. On the day of hatch, 1200 Cobb male broilers were randomly allocated to 5 groups (8 pens/treatment, 30 birds/pen) including 1) negative control (NC): corn-soybean meal diet; 2) positive control (PC): NC + 453 g Stafac20/907 kg feed; 3) Prob 1: NC + 453 g Prob 1/907 kg feed; 4) Prob 2: NC + 453 g Prob 2/907 kg feed; and 5) Synb: NC + 453 g Synb/907 kg feed. One day after placement, birds were challenged by a coccidia vaccine to induce NE. Feed intake and body weights were measured on day 8 (NE onset) and end of starter (day 14) and grower (28) periods. On day 8, the small intestines of 3 birds/pen were examined for NE lesions. Ileal mucosal scrapings from one bird/pen were collected on day 8 and day 28 to profile the microbiota using 16S rRNA sequencing. Data were analyzed in JMP or QIIME 2 and significance between treatments identified by LSD or linear discriminant analysis effect size (P < 0.05). The Synb group significantly lowered NE lesion scores on day 8 and reduced day 0-14 mortality by 50% compared with NC. FCR was significantly better in all the groups, whereas ADG was higher in PC, Synb, and Prob 2 groups compared with NC from day 0 to day 28. Lower lesion scores in the Synb group were accompanied by lower relative abundance of Alistipes, ASF356, Faecalibaculum, Lachnospiraceae UCG-001, Muribaculum, Oscillibacter, Parabacteroides, Rikenellaceae RC9 gut group, Ruminococcaceae UCG-014, and Ruminiclostridium 9 compared with NC on day 8. On day 28, relative abundance of Lactobacillus was lower, whereas abundance of Bacteroides, Barnesiella, Butyricicoccus, CHKCI001, Eisenbergiella, Eubacterium hallii group, Helicobacter, Ruminococcaceae UCG-005, Ruminococcus torques group, and Sellimonas was significantly higher in the NC birds than in the Synb and Prob 2 groups. Collectively, these data indicate that during a subclinical naturally occurring NE, supplementation of specific additives could be effective in reducing intestinal lesions and mortality, and improving performance potentially through developing a signature microbial profile in the intestinal mucosal layer.

Effect of Probiotics and Multi-Component Feed Additives on Microbiota, Gut Barrier and Immune Responses in Broiler Chickens During Subclinical Necrotic Enteritis.

The withdrawal of antibiotic growth promoters from poultry feed has increased the risk of necrotic enteritis (NE) outbreaks. This study examined the effects of a probiotic (PROB) or probiotic/prebiotic/essential oil supplement (PPEO) during a subclinical NE challenge. On day (d) of hatch, 960 male broilers were randomized to four groups (8 pens/treatment, 30 birds/pen) including (1) negative control (NC): corn-soybean meal diet; (2) positive control (PC): NC + 20 g Virginiamycin/ton diet; (3) NC + 227 g PROB/ton diet; and (4) NC + 453 g PPEO/ton diet. One d after placement, birds were challenged by a coccidia vaccine to induce NE. Feed intake and body weights were measured on d 8 (NE onset) and end of each feeding period. On d 8, the small intestines of three birds/pen were examined for NE lesions. Jejunum samples and ileal mucosal scrapings from one bird/pen were respectively collected to measure mRNA abundance (d 8 and d 14) and profile the microbiota (d 8 and d 42). Data were analyzed in JMP or QIIME 2 and significance between treatments identified by LSD (P < 0.05). PROB and PPEO had significantly lower mortality (d 0-14) and NE lesion scores compared to NC. Feed conversion ratio was significantly lower in PC, PROB, and PPEO, while average daily gain was higher in PPEO and PC groups compared to NC from d 0-42. On d 8 and d 14, mRNA abundance of claudin-3 was higher in PPEO compared to NC. On d 14, compared to NC, mRNA abundance of sIgA and PGC-1α in PROB and PPEO were lower and higher, respectively. Compared to NC, PPEO increased mTOR abundance on d 14. On d 8, relative abundance of Clostridium sensu stricto 1, Ruminiclostridium9, Prevotellaceae, Prevotellaceae UCG-014, ASF356, and Muribaculaceae was higher in NC compared to PPEO and PROB, while Lactobacillus was lower in NC. Escherichia-Shigella had higher abundance in PC compared to PPEO and PROB. Collectively, these data indicate that during a subclinical naturally occurring NE, supplementation of PROB or PPEO supports performance and reduces intestinal lesions, potentially through modifying tight junction proteins, gut microbiota, immune responses, and cell metabolism.