An ammonia gas (NH₃) generator was developed to maintain a set concentration of ammonia gas in a controlled environment chamber to study poultry physiological responses to sustained elevated levels of ammonia gas. The goal was to maintain 50 parts per million (ppm) of ammonia gas in a 3.7 m × 4.3 m × 2.4 m (12 ft × 14 ft × 8 ft) controlled environment chamber. The chamber had a 1.5 m³/s (3000 cfm) recirculation system that regulated indoor temperature and humidity levels and a 0.06 m³/s (130 cfm) exhaust fan that exchanged indoor air for fresh outdoor air. The ammonia generator was fabricated by coupling ultrasonic humidifiers with an Arduino-based microcontroller and a metallic oxide MQ-137 ammonia gas sensor. Preliminary evaluation under steady conditions showed the average MQ-137 gas sensor accuracy was within 1.4% of the 65.4 ppm concentration measured using a highly accurate infrared gas analyzer. Further evaluation was performed for a setpoint concentration of 50 ppm where ammonia generator reservoirs were filled with 2% or 10% ammonia liquid. For the system tested, it was found that two generators operating at the same time filled with 3.8 L (1.0 gallon) of 2% ammonia cleaning liquid each (7.6 L or 2.0 gallons total) could maintain a gas level of 49.45 ± 0.79 ppm in the chamber air for a duration of 30 h before refilling was required. One generator filled with 3.8 L of 10% ammonia cleaning liquid could maintain 51.24 ± 1.53 ppm for 195 h. Two ammonia generators were deployed for a six-week animal health experiment in two separate controlled environment chambers. The two ammonia generators maintained an average ammonia concentration of 46.42 ± 3.81 ppm and 45.63 ± 4.95 ppm for the duration of the experiment.

Avian necrotic enteritis (NE) is an infectious disease that impacts poultry worldwide causing economic losses. Discontinued use of antimicrobial growth promoters has been associated with high incidence of the disease, which has led to a necessity for finding new therapeutic alternatives. Flavonoids are polyphenolic compounds that have been studied for their health-promoting properties in animals and humans. This study presents a flavonoid-rich corn (PennHFD), as a potential alternative for ameliorating NE in broiler chickens. The effect of a diet formulated with PennHFD was compared to a diet based on commercially available corn in chickens subjected to a controlled challenge of NE based on a co-infection of Eimeria maxima and Clostridium perfringens. Birds fed on the PennHFD-based diet had lower incidence of intestinal lesions (P = 0.048), higher body weight gain (P < 0.01), lower feed conversion ratio (P < 0.01), and lower mortality rates (P = 0.023) compared to the control diet. Therefore, we concluded that the inclusion of the high-flavonoid PennHFD reduces the severity of an experimental challenge of NE in broiler chickens.

Objective: Probiotics have been used in poultry production to improve the performance and health of chickens raised without antibiotics. The combination of different probiotic strains has been used with the hope of conferring multiple benefits to the host. However, the inclusion of several strains does not necessarily boost benefits. There is a lack of studies that compare the efficacy of multi-strain probiotics to their individual components. In this study, the effects of a Bacillus-based probiotic product mix containing B. coagulans, B. licheniformis, B. pumilus, and B. subtilis against Clostridium perfringens were tested in vitro using a co-culture method. The individual strains and different combinations of the strains used in the product were also tested against C. perfringens. Results: The probiotic product mix tested in this study did not show effects against C. perfringens (P = 0.499). When tested individually, the strain of B. subtilis was the most efficient strain to decrease C. perfringens concentrations (P ≤ 0.01), and the addition of other Bacillus species strains significantly decreased its efficacy against C. perfringens. We concluded that the probiotic mix of Bacillus strains used in this study (B. coagulans, B. licheniformis, B. pumilus and B subtilis) was not effective in decreasing C. perfringens concentrations in vitro. However, when deconstructing the probiotic, the strain of B. subtilis alone or combined with the strain of B. licheniformis were effective against C. perfringens. This suggests that the anticlostridial properties of the particular strains of Bacillus used in this study were negatively affected when combined with other Bacillus spp. strains.

<!-- *** Custom HTML *** --> The effect of aspirin on intestinal lesions was evaluated in birds undergoing an experimental challenge with Clostridium perfringens as part of a model for inducing subclinical necrotic enteritis (SNE). Broilers were raised on clean wood shavings and randomly assigned to three treatments: Uninfected (U), Infected (I), and Infected + Aspirin (I+A; 0.025% acetylsalicylic acid in drinking water during days 21–25). Birds in the I and I+A groups were gavaged with Eimeria maxima on day 18 and their feed was inoculated with C. perfringens (1 × 10⁹ CFU/bird) during days 23–25. On day 26, birds were euthanatized, intestinal lesions were evaluated, and intestinal tissue was collected for qPCR assessment of genes thought to be involved in the immune response to SNE: IL-1β, IL-10, MMP-2, and MMP-7. Birds in the I+A group had more-severe and numerous lesions compared to the I group. For all genes except MMP-2, expression was upregulated in the I group compared to the U group, but did not differ between the I and I+A groups. These results indicate that aspirin exacerbated the intestinal lesions associated with this disease. Aspirin could play a role in the development of a reliable and consistent model for the induction of necrotic enteritis under experimental settings.

Bacterial chondronecrosis with osteomyelitis (BCO) is a septic necrosis of the skeletal system of unknown origin and an important cause of lameness in broiler chickens. Epithelial inflammation has been proposed as an avenue for bacterial translocation leading to BCO. We evaluated the effect of subclinical necrotic enteritis (SNE), an intestinal inflammatory event, with the development of BCO. In each of two experiments, chickens were divided into three treatments: (1) SNE challenge, including both dietary (wheat- and fish-based diet) and intestinal pathogenic challenges (Eimeria maxima and Clostridium perfringens), (2) dietary challenge only, and (3) control diet (corn-and soy-based diet). Floor ramps were introduced as part of an established method for increasing the frequency of BCO. The efficacy of the SNE challenge was corroborated by necropsy evaluation of a representative sample of the population. At the end of each experiment, all birds were evaluated for BCO. A high incidence of BCO was found, even in birds with no external signs of lameness. However, the incidence of BCO was not correlated with the intestinal challenge. Conclusions: under the conditions used in these studies, a treatment that is associated with severe damage to the intestinal mucosa does not change the incidence of BCO in broiler chickens.

The search for alternative therapies to antimicrobial growth promoters (AGP) in poultry production has gained momentum in the past years because of consumer preference and government restrictions on the use of AGP in animal production. Flavonoids are plant-derived metabolites that have been studied for their health-promoting properties that could potentially be used as an alternative to AGP in poultry. In a previous study, we showed that the inclusion of a flavonoid-rich corn cultivar (PennHFD1) in the diet improved the health of broilers undergoing necrotic enteritis. However, the mechanisms of action by which the PennHFD1-based diet ameliorated necrotic enteritis are unknown. This study describes the microbial diversity and composition of the jejunum and ileum of chickens co-infected with Eimeria maxima and Clostridium perfringens and treated with a high-flavonoid corn-based diet. Luminal content and mucosal samples from the jejunum and ileum were collected for DNA extraction, 16S rRNA amplicon sequencing and data analyses. The infection model and the dietary treatments significantly changed the alfa diversity indices (Mucosal samples: ASVs, P = 0.04; Luminal content samples: ASVs, P = 0.03), and beta diversities (Mucosal samples: P < 0.01, Luminal content: P < 0.01) of the ileal samples but not those of the jejunal samples. The microbial composition revealed that birds fed the high-flavonoid corn diet had a lower relative abundance of C. perfringens compared to birds fed the commercial corn diet. The treatments also changed the relative abundance of other bacteria that are related to gut health, such as Lactobacillus. We concluded that both the infection model and the dietary high-flavonoid corn changed the broilers’ gut microbial diversity and composition. In addition, the decrease in the relative abundance of C. perfringens corroborates with a decrease in mortality and intestinal lesions due to necrotic enteritis. Collecting different segments and sample types provided a broader understanding of the changes in the gut microbiota among treatments.