Exploring the complexities of poultry respiratory microbiota: colonization, composition, and impact on health.

An accurate understanding of the ecology and complexity of the poultry respiratory microbiota is of utmost importance for elucidating the roles of commensal or pathogenic microorganisms in the respiratory tract, as well as their associations with health or disease outcomes in poultry. This comprehensive review delves into the intricate aspects of the poultry respiratory microbiota, focusing on its colonization patterns, composition, and impact on poultry health. Firstly, an updated overview of the current knowledge concerning the composition of the microbiota in the respiratory tract of poultry is provided, as well as the factors that influence the dynamics of community structure and diversity. Additionally, the significant role that the poultry respiratory microbiota plays in economically relevant respiratory pathobiologies that affect poultry is explored. In addition, the challenges encountered when studying the poultry respiratory microbiota are addressed, including the dynamic nature of microbial communities, site-specific variations, the need for standardized protocols, the appropriate sequencing technologies, and the limitations associated with sampling methodology. Furthermore, emerging evidence that suggests bidirectional communication between the gut and respiratory microbiota in poultry is described, where disturbances in one microbiota can impact the other. Understanding this intricate cross talk holds the potential to provide valuable insights for enhancing poultry health and disease control. It becomes evident that gaining a comprehensive understanding of the multifaceted roles of the poultry respiratory microbiota, as presented in this review, is crucial for optimizing poultry health management and improving overall outcomes in poultry production.

Research Note: Non-invasive urate sphere corticosterone metabolite as a novel biomarker for heat stress in poultry.

Heat stress (HS) poses a critical concern for the global poultry industry. To better understand its impact and mechanism on poultry species, as well as to develop effective HS mitigation options, it is crucial to validate reliable and effective biomarkers of HS. In this study, broiler chickens (96 mixed-sex, 1-day-old) were randomly assigned to 2 environmental conditions: HS at 31°C for 8 h/d versus a thermoneutral condition (TN) at 24°C ± 0.2 throughout the day, from d 21 to d 28. After 7 d of cyclic HS, blood serum samples and the urate sphere (the white portion of the chicken excreta) were collected to measure specific HS biomarkers, including corticosterone, heat shock protein 70 (HSP70), Triiodothyronine (T3), and urate sphere corticosterone metabolite. The results showed that the concentrations of HSP70, T3, and serum corticosterone were not significantly different (P > 0.05) between the heat-stressed and thermoneutral birds in this study. However, heat-stressed birds recorded higher (P < 0.01) concentration (pg/mL) of urate corticosterone metabolite compared to TN birds. Data presented here thus provide preliminary evidence suggesting the use of non-invasive urate sphere corticosterone metabolite as a heat stress biomarker in poultry.

An investigation of the effect of folic acid and its delivery routes on broiler chickens' hatch and growth performance, blood biochemistry, anti-oxidant status, and intestinal morphology.

This study investigated the effect of folic acid (FA) and its delivery routes (in-feed or in ovo) on broiler chicken's hatch and growth performance, blood biochemistry, anti-oxidant status, and intestinal morphology. A total of 1,860 Cobb 500 hatching eggs were incubated for 21 d. On day 12 of incubation, viable eggs were randomly allotted to four groups: the noninjected group, in ovo saline (injected with 0.1 mL/egg of saline solution), in ovo FA 1 (injected with 0.1 ml FA containing 0.1 mg/egg; FA1), and in ovo FA 2 (injected with 0.1 ml FA containing 0.15 mg/egg). All in ovo treatments were delivered via the amnion. At hatch, chicks were re-allotted to five new treatment groups: FA1, FA2, in-feed FA (FA 3; 5mg/kg in feed), in-feed bacitracin methylene disalicylate (BMD; 55 mg/kg in feed), and negative control (NC; corn-wheat-soybean diet) in 6 replicate pens (22 birds/pen) and raised in starter (days 0 to14), grower (days 15 to 24), and finisher (days 25 to 35) phases. Hatch parameters were assessed on day 0, and body weight and feed intake (FI) were determined weekly. On day 25, 1 bird/cage was euthanized, immune organs weighed, and intestinal tissues harvested. Blood samples were collected for biochemistry and anti-oxidant (Superoxide dismutase-SOD and Malondialdehyde-MDA) analysis. Data were analyzed in a randomized complete block design. While FA1 and FA2 decreased (P < 0.001) hatchability in a dose-dependent manner, FA2 caused a 2% increase (P < 0.05) in average chick weight compared to the noninjected group. Compared to the BMD treatment, FA3 decreased (P < 0.05) average FI across all feeding phases. At the end of the trial on day 35, FA2 had similar feed conversion ratio as the BMD treatment while recording less (P < 0.001) FI. FA1 and FA2 recorded a tendency (P < 0.1) to increase MDA levels and SOD activity by 50% and 19%, respectively, compared to the NC treatment. Compared to NC treatment, FA2 increased (P < 0.01) villus height, width, and villus height to crypt depth ratio in the duodenum, and villus width in the jejunum. Besides its negative effect on hatchability, FA2 may help improve embryonic development and anti-oxidant status in broiler chickens.

Folic acid (FA) is an essential vitamin with anti-oxidant and growth-promoting properties. Notwithstanding, studies have shown that, like other bioactive substances, its efficacy can be affected by dosage and delivery routes. However, the in ovo delivery of bioactive substances directly to the developing embryo continues to show promising results in poultry production. Consequently, this study evaluated the effect of two doses of in ovo-delivered folic acid on selected broiler chicken performance indicators. The results revealed that both doses of in ovo-delivered FA decreased hatchability. However, the higher dose increased hatched chick's weight. The higher dose of the in ovo-delivered FA also increased broiler chicken intestinal morphology while showing a tendency to improve anti-oxidant activities.

An evaluation of the thermoregulatory potential of in ovo delivered bioactive substances (probiotic, folic acid, and essential oil) in broiler chickens.

Mitigating the negative effects of heat stress (HS) is a critical challenge for the global poultry industry. This study evaluated the thermoregulatory potential of 3 in ovo delivered bioactive substances using selected gut health parameters. Eggs were incubated and allotted to 5 groups, and respective bioactive substances delivered. These groups included-the noninjected, in ovo saline, in ovo folic acid (FA), in ovo probiotics (P), and in ovo essential oil (EO). At hatch, chicks were assigned to 5 new posthatch treatment combinations, including A) Negative control (NC)-noninjected eggs offered a basal corn-wheat-soybean diet, B) Antibiotics-NC + 0.05% bacitracin methylene disalicylate, C) In ovo FA-eggs injected with FA + NC diet, D) In ovo probiotics-eggs injected with probiotics + NC diet, E) In ovo + in-water EO-eggs injected with EO and supplied EO via drinking water + NC diet. Birds were raised for 28 d in 8 replicate cages/treatment (6 birds/cage) and exposed to either a thermoneutral (24°C ± 0.2) or HS challenge (31°C) condition from d 21 to d 28. The in ovo delivered FA and EO treatments reduced (P < 0.001) hatchability by at least 26% compared to NC. Induced HS reduced (P < 0.001) total plasma protein, total antioxidant capacity, and villus width in the duodenum and jejunum. Independent of HS and compared to NC, the in ovo + in-water EO treatment resulted in (P < 0.05) at least a 15% increase in villus height: crypt depth across the 3 gut sections. The in ovo + in-water EO treatment also increased the relative mRNA expression of intestinal barrier-related genes (Claudin1,3,4, Occludin, Zona occludens-2, and Mucin 2). Under HS, the in ovo + in-water EO treatment recorded a 3.5-fold upregulation of amino acid transporter gene (SLC1A1), compared to NC. Subject to further hatchability optimization, the in ovo + in-water delivery of EO show potential to afford broiler chicken thermotolerance.

The effect of Bacillus subtilis and its delivery route on hatch and growth performance, blood biochemistry, immune status, gut morphology, and microbiota of broiler chickens.

This study evaluated the effect of probiotics (Bacillus subtilis fermentation extract) and its delivery route (in-feed or in ovo) on hatch and growth performance, blood biochemistry, immune status, gut morphology, and microbiota of broiler chickens. Hatching eggs were incubated for 21 d. On d 12, viable eggs were randomly allotted to 4 groups: the noninjected, in ovo saline (S), in ovo Bacillus subtilis 1 (P1), and in ovo Bacillus subtilis 2 (P2). On d 18, S, P1, and P2 groups received 0.2 mL saline diluent, 10 × 106, and 20 × 106 CFU of the bacterium via the amnion, respectively. At hatch, chicks were re-allotted to 5 new treatment groups: P1, P2, 0.005% in-feed Bacillus subtilis extract (P3), 0.05% in-feed bacitracin methylene disalicylate (BMD,), and corn-wheat-soybean diet negative control (NC) in 9 replicate pens (22 birds/pen) and raised for 35 d. Hatch parameters were assessed on d 0, and growth performance indices measured weekly. On d 25, 1 bird/cage was euthanized, and samples collected for further analysis. Data were analyzed by generalized linear model. Treatments S and P2 recorded higher (P = 0.01) chick BW/ Egg Weight values compared to the non-injected eggs. P3 and P2 reduced (P = 0.02) FI at week 5 compared to the NC treatment. However, no change in average body weight gain (ABG) and feed conversion ratio (FCR) were observed during the same period. At d 35, while BMD treatment showed a tendency (P = 0.09) to increase FI compared to the NC treatment, ABG and FCR were similar for all treatments. Blood sodium and chloride levels were increased (P < 0.05) by the BMD treatment compared to the NC treatment. Compared to other treatments, BMD and P3 treatments increased (P < 0.001) jejunal and ileal villus height to crypt depth ratios, respectively. However, P1 and P2 increased (P < 0.001) villus height to crypt depth ratio in the duodenum compared to NC treatment. Treatments did not affect gut microbial diversity; however, BMD treatment increased (P < 0.05) the proportion of bacteria in the genus Enterococcus in the ileum and reduced (P < 0.05) the proportion of bacteria in the genus Streptococcus in the ceca. All probiotics treatments (irrespective of route and dose) reduced (P < 0.001) the levels of serum IgG compared to the NC treatment. However, P1 and P2 had the lowest numerical decrease in serum IgG concentrations, suggesting that Bacillus subtilis (especially in ovo delivered) might provide broiler chickens with better immunological protection by neutralizing pathogenic organisms that could result in the production of natural antibodies.

Biomarkers of heat stress and mechanism of heat stress response in Avian species: Current insights and future perspectives from poultry science.

Heat stress remains a challenge for all living organisms on the globe, especially in the summer months. Heat stress is one of the most critical environmental stressors impairing the welfare and productivity of avian species, especially poultry. Heat stress evokes adverse effects, which include but are certainly not limited to immunosuppression, compromised gut health, oxidative damage, and reduced growth performance in birds. It is now increasingly evident that accurate measurements of heat stress before adverse effects are manifested signify a critical step towards developing effective heat stress mitigative strategies. Despite the growing body of knowledge on the impacts of heat stress on avian species and possible mitigative strategies, a comprehensive overview of the methods and biomarkers available to evaluate the occurrence of heat stress is lacking in the literature. Accordingly, this review provides a detailed overview of the physiological mechanisms underlying heat stress in birds. Additionally, the merits and limitations of current invasive and non-invasive measures of heat stress are highlighted. Furthermore, justifications for the adoption of novel non-invasive biomarkers of heat stress are also presented. Practical recommendations on the use of these biomarkers are equally offered. There is no doubt that the information provided herein would benefit both avian physiologists and ecologists by sparking the much-needed conversation in the quest for reliable heat stress biomarkers.

Microbiota and Transcriptomic Effects of an Essential Oil Blend and Its Delivery Route Compared to an Antibiotic Growth Promoter in Broiler Chickens.

This study evaluated the effect of the delivery of a commercial essential oil blend containing the phytonutrients star anise, cinnamon, rosemary, and thyme oil (via different routes) on broiler chickens' ileal and ceca microbiota and liver transcriptome compared to an antibiotic growth promoter. Eggs were incubated and allocated into three groups: non-injected, in ovo saline, and in ovo essential oil. On day 18 of incubation, 0.2 mL of essential oil in saline (dilution ratio of 2:1) or saline alone was injected into the amnion. At hatch, chicks were assigned to post-hatch treatment combinations: (A) a negative control (corn-wheat-soybean diet), (B) in-feed antibiotics, (C) in-water essential oil (250 mL/1000 L of drinking water), (D) in ovo saline, (E) in ovo essential oil, and (F) in ovo essential oil plus in-water essential oil in eight replicate cages (six birds/cage) and raised for 28 days. On days 21 and 28, one and two birds per cage were slaughtered, respectively, to collect gut content and liver tissues for further analysis. Alpha and beta diversity differed significantly between ileal and ceca samples but not between treatment groups. In-feed antibiotic treatment significantly increased the proportion of specific bacteria in the family Lachnospiraceae while reducing the proportion of bacteria in the genus Christensenellaceae in the ceca, compared to other treatments. Sex-controlled differential expression of genes related to cell signaling and tight junctions were recorded. This study provides data that could guide the use of these feed additives and a foundation for further research.

Dietary grape pomace - effects on growth performance, intestinal health, blood parameters, and breast muscle myopathies of broiler chickens.

The search for alternatives to antibiotics in poultry production is still on-going and has been directed towards investigation of the efficacy of different potential alternatives. However, it is important that the sought alternatives are cost-efficient and have no negative impact on meat quality, for ease of adoption and profit maximization. This study aimed at exploiting an agro-industrial waste, grape pomace (GP) as an alternative to in-feed antibiotics and assessing the effects on growth, intestinal morphology, ceca microbiota, ceca short-chain fatty acid (SCFA) concentration, blood biochemical parameters, and breast muscle myopathies of broiler chickens. A total of 576 one-day-old Cobb-500 broiler chicks were randomly allotted to 3 dietary treatments - Negative control (NC, a corn-wheat soybean-based diet), NC + 0.05% bacitracin methylene disalicylate (BMD), and NC + 2.5% GP. Each treatment was assigned to 8 replicate pens with 25 birds per pen. Body weight (BW), feed intake (FI), and feed conversion ratio (FCR) were determined weekly. On d 36, 2 chickens/pen were euthanized for measuring blood biochemical parameters, ceca SCFA, and ceca microbiota. White striping (WS) and wooden breast (WB) incidence were assessed in 4 chickens/pen on d 42. The GP diet increased (P < 0.05) average FI throughout the feeding phases compared to the other treatments, but overall FCR was similar. Birds in the GP treatment had higher (P < 0.05) villus height (VH) and increased VH:crypt depth ratio in the duodenum and jejunum compared to other treatments. The level of ceca SCFA and the incidence of WS and WB was the same for all treatments. Plasma Ca and P were significantly higher (P < 0.05) in birds fed GP and BMD, compared to the NC. Birds in the GP treatment had significantly reduced (P < 0.05) plasma aspartate transaminase than other treatments. Birds receiving GP had a higher (P < 0.05) relative abundance of the phylum Bacteroidetes and reduced (P < 0.05) Firmicutes compared to other treatments. The relative abundance of Bacteroides and Lactobacillus genera were higher (P < 0.05) among birds fed GP compared to other treatments. Inclusion of 2.5% GP in broiler chicken diets improved gut morphology and modified the cecal bacterial community and blood biochemical profiles with no adverse effect on growth performance and meat quality.

Essential Oil Delivery Route: Effect on Broiler Chicken's Growth Performance, Blood Biochemistry, Intestinal Morphology, Immune, and Antioxidant Status.

This study evaluated the effect of an essential oil blend and its delivery routes on broiler chicken growth performance, blood biochemistry, intestinal morphology, and immune and antioxidant status. Eggs were incubated and allotted to 3 groups: non-injected group, in ovo saline group, and in ovo essential oil group. On day 18 of incubation, essential oil in saline or saline alone was injected into the amnion. At hatch, chicks were assigned to post-hatch treatment combinations (1) in ovo essential oil + in-water essential oil (in ovo + in-water EO); (2) in ovo essential oil (in ovo EO); (3) in ovo saline; (4) in-water essential oil; (5) in-feed antibiotics (Bacitracin methylene disalicylate) and (6) a negative control (NC; corn-wheat-soybean diet) in 8 replicate cages (6 birds/cage) and raised for 28 day. The in ovo EO group reduced (p < 0.05) chick length and hatchability, all groups recorded no difference in growth performance at 0-28 day. The in ovo + in-water EO treatment reduced (p < 0.05) blood creatine kinase and aspartate aminotransferase levels whilst increasing (p < 0.05) total antioxidant capacity in birds. The in ovo + in-water delivery of EO might represent a potential antibiotic reduction strategy for the poultry industry but more research is needed to address the concern of reduced hatchability.

Effect of organic acids-essential oils blend and oat fiber combination on broiler chicken growth performance, blood parameters, and intestinal health.

This study evaluated the effect of organic acids-essential oils blend with or without oat hulls (OH) on growth performance, organ weights, blood parameters, gut morphology, microbiota, and short-chain fatty acids (SCFA) in broiler chickens. Day-old broiler chickens were randomly allocated to 4 dietary treatments consisting of 1) a corn-soybean meal-wheat based diet (BAS), 2) BAS + 0.05% bacitracin methylene disalicylate (BMD), 3) BAS + protected organic acids-essential oils at 300 g/1,000 kg of feed (OE), and 4) BAS + protected organic acids-essential oils at 300 g/1,000 kg of feed + 3% OH (OEOH), in 8 replicate groups. Feeding was in starter (d 0 to 14), grower (d 14 to 24), and finisher (d 24 to 36) phases. Body weight (BW), feed intake (FI), feed conversion ratio (FCR), and mortality were determined weekly. On d 36, 8 chickens per treatment were sampled for blood biochemistry, organ weights, cecal SCFA production, and microbiota. Treatments had no effect on FI and FCR at all phases. Both OE and OEOH treatments reduced (P < 0.001) the body weight gain of birds at the starter phase. Birds fed the OEOH treatment had higher (P < 0.001) gizzard weight, while those offered the BMD diet showed a tendency (P = 0.08) to have higher cecal weight. Birds in the OEOH treatment recorded increased ileal villus height and villus height-to-crypt depth ratio, as well as reduced duodenal crypt depth, while birds in the OE treatment had increased jejunal villus height and villus height-to-crypt depth ratio. Both OEOH and OE treatments increased the number of goblet cells produced in the duodenum and jejunum. Treatments had no effect on SCFA concentrations. Birds in the OE treatment recorded the lowest concentration of blood urea (P = 0.05) and cholesterol (P < 0.05). Both OE and OEOH treatments increased (P < 0.05) the relative abundance of potentially beneficial bacteria in the genus Firmicutes_unclassified, Ruminococcus, Turicibacter, and Erysipelotrichaceae_unclassified, while reducing (P < 0.001) the relative abundance of potentially harmful Coprobacillus. Conclusively, both protected organic acids-essential oils blend and its combination with oat fibers show potential as tools to achieve antibiotics reduction in broiler production.

Bacillus subtilis delivery route: effect on growth performance, intestinal morphology, cecal short-chain fatty acid concentration, and cecal microbiota in broiler chickens.

As the poultry industry recedes from the use of antibiotic growth promoters, the need to evaluate the efficacy of possible alternatives and the delivery method that maximizes their effectiveness arises. This study aimed at expounding knowledge on the effect of the delivery method of a probiotic product (Bacillus subtilis fermentation extract) on performance and gut parameters in broiler chickens. A total of 450 fertile eggs sourced from Cobb 500 broiler breeders were randomly allotted to 3 groups: in ovo probiotic (n = 66), in ovo saline (n = 66), and noninjection (n = 200) and incubated for 21 d. On day 18.5 of incubation, 200 µL of either probiotic (10 × 106 cfu) or saline was injected into the amnion. At hatch, chicks were reallotted to 6 new treatment groups: in ovo probiotic, in ovo saline, in-feed antibiotics, in-water probiotic, in-feed probiotics, and control (corn-wheat-soybean diet) in 6 replicate cages and raised for 28 d. Of all hatch parameters evaluated, only percentage pipped eggs was found significant (P < 0.05) with the noninjection group having higher percentage pipped eggs than the other groups. Treatments did not affect the incidence of necrotic enteritis on day 28 (P > 0.05). Irrespective of the delivery method, the probiotic treatments had no significant effect on growth performance. The ileum villus width of the in ovo probiotic treatment was 18% higher than the in ovo saline group (P = 0.05) but not statistically higher than other groups. The jejunum villus height was 23% higher (P = 0.000) in the in ovo probiotic group than in the control group. There was no effect of treatment on total cecal short-chain fatty acid concentration and cecal gut microbiota composition and diversity (P > 0.05), although few unique bacteria differential abundance were recorded per treatment. Conclusively, although probiotic treatments (irrespective of the delivery route) did not affect growth performance, in ovo delivery of the probiotic product enhanced intestinal morphology, without compromising hatch performance and gut homeostasis.