This study identified the optimal multi-enzyme dose rate at three energy levels based on the production performance of broiler chickens. A 42-day grow out trial was conducted using 576 day-old mixed-sex ROSS308 broiler chickens in a 3 × 4 factorial arrangement in a completely randomized design. Diets consisting of three metabolizable energy (ME) levels: standard energy (STD), 150 kcal/kg energy reduction (STD-150), and 200 kcal/kg energy reduction (STD-200), were cross factored with four multi-enzyme inclusion levels (0, 350, 700, and 1000 g/ton). The average daily feed intake and feed conversion ratio increased linearly (p < 0.001) as the dietary ME was reduced, and the multi-enzyme addition improved the feed conversion ratio (p < 0.05) and mitigated the negative effect of the reduced energy diets (RED) on feed intake and feed conversion ratios. Carcass composition, organ weights, and meat quality were not affected by the experimental diets. The RED decreased abdominal fat weight (p < 0.05). Total ash, calcium, and phosphorous contents of the tibia bone were improved (p < 0.04) when the RED were supplemented with the multi-enzyme. Super-dosing multi-enzymes in RED mitigates the negative effect of ME reduction on growth performance while maintaining organ development and meat quality and improving bone mineral content.

Considerable research has been conducted into the efficacy of individual probiotics in broiler production, however information on the most effective combinations of synergistic Bacillus probiotic is lacking. This study investigated the impact of different Bacillus strain combinations in broiler chickens, as well as in vitro enzyme production. In experiment one, a total of 576 Ross 308 broilers at 1 d old were grown for 21 d across 6 treatments of maize-soybean diets (n = 12 pens per treatment) to compare three different strain combinations (formulation 1 [F1]: 3 strains Bacillus amyloliquefaciens; F2: Bacillus coagulans and 2 strains B. amyloliquefaciens; F3: B. coagulans, Bacillus licheniformis and 2 strains B. amyloliquefaciens; F5: Bacillus subtilis, B. licheniformis and 2 strains B. amyloliquefaciens), positive control (PC), and a negative control antibiotic treatment group (NC). In Exp. 2, a total of 360 one-day-old ROSS308 broilers were used to test five treatments (n = 9) including PC, NC, F1 and F5 (selected from Exp. 1), and F4 (Bacillus pumilis and 2 strains B. amyloliquefaciens) in a maize-soybean diet. B. amyloliquefaciens F1 demonstrated a significant improvement in feed conversion ratio (FCR) compared to F2 at d 14 (1.49 vs 2.10; P = 0.038) and the body weight (BW) at d 21 (847.0 g vs 787.4 g) compared to other combinations (P = 0.027). The FCR at d 21 tended to be lower in birds fed F1 (1.46 vs 1.66) compared to the control (P = 0.068). Probiotic treatments had significantly improved nutrient digestibility compared to the PC and NC. Also, probiotic treatments supported the growth of Streptococcus, a common commensal genus and reduced the abundance of genera that correlated with low weight gain such as Akkermansia. Experiment two revealed that F4 improved FCR (P < 0.001) and BW at 28 d (P = 0.014). In vitro testing showed a high production of protease and amylase by Bacillus. Thus, the addition of Bacillus probiotics, particularly containing B. amyloliquefaciens strains and Bacillus pumilus, into the diet of broiler chickens improves production performance, nutrient digestibility, and allows the proliferation of beneficial gut microbiota.

Optimizing gut health has a large impact on nutrient digestibility and bioavailability, and super-dosing feed enzymes may be one solution to achieve this. A 42-day grow-out trial was conducted using 192 Ross 308 broilers to determine if super-dosing Natuzyme at 0 g/t, 350 g/t, 700 g/t, and 1000 g/t dose rates could improve the gut morphology, alter the cecal microbial profile, enhance bone mineralization, and improve nutrient digestibility of a wheat–corn–soybean diet (six replicates per treatment, eight birds per pen). One bird per pen was slaughtered at day 42 and gut morphology, cecal microbial profile, and nutrient digestibility were studied. The addition of enzymes tended to increase the villus height in the duodenum, villus height, width, and crypt depth in the jejunum, and villus width and the number of goblet cells in the ileum. Microbial profiling revealed diverse communities; however, they did not significantly differ between treatment groups. Yet, 700 g/t Natuzyme promoted microbes belonging to the genus Romboutsia and Ruminococcus gauvreauii, while 1000 g/t Natuzyme promoted Barnesiella species. The nutrient digestibility demonstrated a significant improvement in all enzyme doses compared to the control. In conclusion, based on the outcomes of this study, a dose rate of 700 g/t Natuzyme is recommended to improve gut morphology and nutrient digestibility, and promote unique microbes which aid in feed efficiency.

Human cutaneous squamous cell carcinomas (SCCs) and actinic keratoses (AK) display microbial dysbiosis with an enrichment of staphylococcal species, which have been implicated in AK and SCC progression. SCCs are common in both felines and canines and are often diagnosed at late stages leading to high disease morbidity and mortality rates. Although recent studies support the involvement of the skin microbiome in AK and SCC progression in humans, there is no knowledge of this in companion animals. Here, we provide microbiome data for SCC in cats and dogs using culture-independent molecular profiling and show a significant decrease in microbial alpha diversity on SCC lesions compared to normal skin ( P ≤ 0.05). Similar to human skin cancer, SCC samples had an elevated abundance of staphylococci relative to normal skin—50% (6/12) had >50% staphylococci, as did 16% (4/25) of perilesional samples. Analysis of Staphylococcus at the species level revealed an enrichment of the pathogenic species Staphylococcus felis in cat SCC samples, a higher prevalence of Staphylococcus pseudintermedius in dogs, and a higher abundance of Staphylococcus aureus compared to normal skin in both companion animals. Additionally, a comparison of previously published human SCC and perilesional samples against the present pet samples revealed that Staphylococcus was the most prevalent genera across human and companion animals for both sample types. Similarities between the microbial profile of human and cat/dog SCC lesions should facilitate future skin cancer research. IMPORTANCE: The progression of precancerous actinic keratosis lesions (AK) to cutaneous squamous cell carcinoma (SCC) is poorly understood in humans and companion animals, despite causing a significant burden of disease. Recent studies have revealed that the microbiota may play a significant role in disease progression. Staphylococcus aureus has been found in high abundance on AK and SCC lesions, where it secretes DNA-damaging toxins, which could potentiate tumorigenesis. Currently, a suitable animal model to investigate this relationship is lacking. Thus, we examined the microbiome of cutaneous SCC in pets, revealing similarities to humans, with increased staphylococci and reduced commensals on SCC lesions and peri-lesional skin compared to normal skin. Two genera that were in abundance in SCC samples have also been found in human oral SCC lesions. These findings suggest the potential suitability of pets as a model for studying microbiome-related skin cancer progression.

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