The microbiome has entered the vernacular of the consumer as well as broiler production and is, therefore, becoming increasingly important to poultry producers to understand. The microbiome is, by ...definition, compositional and relates to how the microbiological organisms within the gut inhabit that ecological niche. The gut is diverse, flexible, and data acquired requires a greater understanding of the host-microbiome axes, as well as advanced bioinformatics and ecology. There are numerous microbial populations that define the gut microbiome; however, there are even more effects that can influence its composition. As management practices vary between producers, documenting these influences is an essential component of beginning to understand the microbiome. This review targets broiler production and concatenates the currently understood compositional ecology of the broiler gastrointestinal tract microbiome as well as its influences.
This study was conducted to evaluate the effects of different dietary inclusion of eucalyptus essential oil (EEO) on growth performance, relative organ weight, cecal microflora, nutrient ...digestibility, serum biochemical parameters, and thigh muscle fatty acid profile in broiler chickens. A total of six hundred 1-day-old male broiler chickens were randomly allocated into 5 treatment groups with 8 replicate pens, and each pen contained 15 birds. The experiment lasted for 42 d. Dietary treatments included corn–soybean meal-based diet supplemented with 0, 250, 500, 750, and 1,000 mg/kg EEO. The results indicated that dietary treatments had no effect on growth performance parameters in the 1 to 10 d period. From day 11 to 24, dietary supplementation of EEO showed a linear decrease in feed conversion ratio (FCR, P < 0.05). From day 25 to 42 and the overall period (1–42 d), broilers fed with different levels of EEO showed a linear increase in body weight gain (BWG) and reduction in feed conversion ratio (linear, P < 0.05). The relative organ weight were unaffected by any of the dietary treatments. With increasing levels of EEO, the cecal Escherichia coli (linear, P = 0.085) count showed a trend in reduction, and the cecal lactic acid bacteria population tended to increase (linear, P = 0.063). The apparent ileal digestibility of ether extract and organic matter were linearly and quadratically increased in response to increasing dietary EEO supplementation (P < 0.05). A trend of linear decrease in total cholesterol in the serum of birds fed with different levels of EEO was recorded (P = 0.074). Eucalyptus essential oil's inclusion increased serum superoxide dismutase linearly but reduced serum malondialdehyde linearly (P < 0.05). Dietary supplementation of EEO affected the fatty acid profile of thigh muscle so that increased the concentrations of total polyunsaturated fatty acids (linear, P < 0.05) and reduced total saturated fatty acid contents (linear, P < 0.05). Taken together, the inclusion of EEO increased BWG and decreased FCR during day 25 to 42 and day 1 to 42, and partially improved cecal microflora balance, nutrient digestibility, antioxidant activity, and thigh muscle fatty acid profile in broiler chickens.
The reduction in antibiotic growth promoter use in poultry, due to antibiotic resistance concerns, has created a need for natural solutions that control enteric pathogens like Salmonella. One of ...these natural feed additives, a select blend of essential oils, fatty acids, and an enterosorbent mineral (NeutraPath), was assessed for its effects on the intestinal colonization of Salmonella enterica serovar Typhimurium PHL2020 isolate (ST-PHL2020) in broiler chickens and ST-PHL2020 virulence gene expression. An in vitro digestion model simulating the pH and enzymatic conditions of 3 gastrointestinal compartments (crop, proventriculus, and intestine) was first used to evaluate the antibacterial effects of NeutraPath on ST-PHL2020. For the in vivo study, day-old male broilers (n = 90) were randomly allocated to 1 of 3 groups: control or NeutraPath supplemented at 0.25 or 0.5%. The dose rates were chosen to enable observable statistical effects during high Salmonella challenge. All groups were challenged with ST-PHL2020 (106 cfu/bird) via oral gavage on day 9. Bacterial load and prevalence of ST-PHL2020 were examined in ceca-cecal tonsils, and intestinal permeability was assessed via serum recovery of fluorescein isothiocyanate dextran (FITC-d) 24 h postchallenge. NeutraPath inhibited (P < 0.05) ST-PHL2020 growth in the in vitro digestion model compared to the control at all concentrations and in all compartments other than NeutraPath 0.25% in the crop. In vivo, NeutraPath 0.25 and 0.5% reduced (P < 0.05) the total cfu recovered and total prevalence of ST-PHL2020 in the ceca. The serum FITC-d levels were also reduced (P < 0.05) by NeutraPath. Further, NeutraPath's effects on ST-PHL2020’s Salmonella pathogenicity island-1 virulence network development were explored via treating ST-PHL2020 at subinhibitory concentration (1 mg/mL) of NeutraPath in vitro. Compared to the control, NeutraPath downregulated ST-PHL2020 hilA and invF mRNA expression, which further blocked expression of key downstream effectors involved in ST-PHL2020 invasion. Collectively, NeutraPath has the potential to reduce ST-PHL2020 intestinal colonization in broilers and preserve intestinal barrier integrity.
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.
The objective of this experiment was to investigate the effects of the dietary soy galactooligosaccharides (GOS), raffinose and stachyose, on performance, gastrointestinal health, and systemic stress ...in young broilers. Birds were fed a GOS-devoid diet based on soy protein isolate (SPI) or the SPI diet with 0.9, 1.8, 2.7, or 3.6% added stachyose and raffinose in a ratio of 4:1 at the expense of corn starch. These 5 treatments were administered to 10 replicate cages of 8 birds. Performance was measured weekly and excreta moisture, N retention, apparent metabolizeable energy, and complete blood cell counts were determined at 14 and 21 d. At 21 d, 2 birds per cage were orally gavaged with fluorescein isothiocyanate-dextran (FITC-d) and serum samples were analyzed for FITC-d as a marker of gut leakage. Additionally, intestinal morphology, crop presumptive lactic acid bacteria (LAB) counts, crop and cecal pH, and cecal microbiota via16S rRNA microbial sequencing were evaluated at 21 d. From 0 to 21 d, feed intake increased linearly (P < 0.01) as dietary GOS increased, whereas BWG increased (P < 0.05) quadratically. Feed conversion ratio increased (P < 0.01) linearly as GOS increased. There were linear increases (P < 0.05) in excreta moisture as dietary GOS increased at 14 and 21 d, as well as dose-dependent responses (P < 0.05) in N retention, AME, and AMEn. There was a quadratic increase (P < 0.05) in crop LAB recovery and a linear decrease (P < 0.01) in ceca pH as GOS increased. At 14 d, a linear increase (P < 0.05) in blood heterophil to lymphocyte ratio was observed as dietary GOS increased. Serum concentrations of FITC-d increased quadratically (P < 0.01) to dietary GOS. Increasing levels of GOS influenced alpha and beta diversities and composition of gut microbiota, including the abundance of Ruminococcus and Bifidobacterium. Results from this trial indicate that soy-derived GOS exert dose-dependent effects on nutrient utilization and intestinal health in young broilers.
Commercial hatch cabinet environments promote replication of microorganisms. These pathogenic or apathogenic microorganisms may serve as pioneer colonizers of the gastrointestinal tract (GIT) of ...poultry. Some of these pioneer colonizers, such as Escherichia coli and Enterococcus spp., are opportunistic pathogens that lead to reduced performance in commercial poultry. Effective hatchery sanitation is imperative to limit contamination of naïve neonatal chicks and poults. Formaldehyde fumigation has been traditionally used to reduce the pathogen load in commercial hatch cabinets. To investigate potential alternatives to formaldehyde fumigation, models to mimic the microbial bloom in a laboratory setting must be utilized. The purpose of the present study was to evaluate the impact of a multispecies environmental challenge model (PM challenge) with and without formaldehyde fumigation during the hatching phase on early performance in broiler chicks. Three experiments were conducted to evaluate microbial contamination in the hatch cabinet environment (air samples, fluff samples), enteric colonization at day-of-hatch (DOH), and 7-day performance. In all experiments, significantly (P < 0.05) more gram-negative bacteria were recovered from the GIT at DOH in the PM challenge control group as compared to the nonchallenged control (NC) group and the formaldehyde-treated group (PM + F). There were no statistical differences in 7-day body weight gain or feed conversion ratio between the PM challenge control group, the NC group or the PM + F group. These data suggest this model could be utilized to evaluate alternatives to formaldehyde fumigation for controlling the microbial load during the hatching phase in a laboratory setting.
Avian pathogenic Escherichia coli (E. coli) is an opportunistic pathogen often introduced to neonatal chicks during the hatching process. This commensal bacterium, particularly as a pioneer colonizer ...of the gastrointestinal tract, can have substantial implications in the rearing of poultry because of reduced flock performance. In order to mimic the effects of the natural bacterial bloom present during the hatch, a seeder challenge model was developed to expose neonatal chicks to virulent E. coli. On day 20 of embryogenesis, selected early hatched chicks (n = 18/hatcher) were briefly removed and sprayed challenged with saline (vehicle) or E. coli at 1 × 107 colony-forming unit (CFU)/chick (exp 1) and 2.5 × 107 CFU/chick (exp 2). These challenged chicks were returned to the hatcher to serve as seeders to transmit the pathogen to the indirect challenged or contact chicks (n = 195/hatcher). For two 7-d experiments, the efficacy of transmission was evaluated via enteric bacterial recovery, body weight gain (BWG), and mortality. For exp 1 and exp 2, significantly (P < 0.0001) more gram-negative bacteria were recovered from the seeder and contact gastrointestinal samples than the negative control samples on day of hatch. In addition, there was a reduction (P < 0.05) in 7-d BWG and significantly (P < 0.0001) higher mortality in the contact-challenged chicks than the negative control chicks in both exp 1 and exp 2. These data suggest that this challenge model could be used to evaluate different methods of controlling the bacterial bloom that occurs in the hatching environment.
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.
After being banned by the European Commission in 2018, the use of formaldehyde as a feed amendment in the United States has come into question. Therefore, this study was conducted to explore ...alternatives to formaldehyde, such as formic acid and monoglycerides, and their effects on poultry production. In total, 1,728 Cobb 700 broilers were randomly assigned to 96-floor pens on day of hatch (18 birds/pen). Using a randomized complete block design (4 blocks), treatments were assigned to pens with blocking based on location within the barn, with the eastern half of the barn designated for digestibility and the western half designated for production (per experiment: 8 control pens and 10 pens per treatment). All diets were based on a negative control (NC), basal diet. Dietary treatments consisted of: NC, NC + 0.25% formalin (F), NC + 0.25 and 0.50% Amasil NA (AML and AMH; 61% formic acid and 20.5% Na-formate), and NC + SILO Health 104L (SILO; mixture of monoglycerides; 0.5% from 0 to 14 d, 0.4% from 14 to 28 d, and 0.2% from 28 to 42 d). Water and feed were provided ad libitum. Performance data were collected during feed changes on d 0, 14, 28, and 42, with digestibility data collected at d 14 (2 per pen) and carcass quality (6 per pen) assessed at d 46 with a randomly selected group of broilers. A one-way ANOVA followed by Dunnett's multiple comparison, where treatments were evaluated against F were conducted using JMP 14.0 (P ≤ 0.05). Main effect of treatment was significant for performance, nutrient digestibility, and carcass quality. Differences in body weight and ADG were observed from d 14 to d 28, resulting in a trending improvement in lysine digestibility on d 14 and carcass quality on d 46 of birds fed AML and AMH in comparison to those fed F (P < 0.05). Whereas birds fed SILO had reduced digestibility of methionine on d 14 and a decrease in meat quality on d 46 in comparison to those fed F (P < 0.05). Therefore, Amasil NA at 0.25 or 0.50% may be an effective alternative to formaldehyde as a feed amendment for poultry production.
This study aimed to evaluate the fate and dissemination of Salmonella Reading (SR) in turkeys using an oral gavage challenge model. One hundred twenty-eight-week-old commercial turkey hens were moved ...from commercial production to research facilities. Upon arrival, a combination of enrofloxacin, 10 mg/kg, and florfenicol, 20 mg/kg, were orally administered sequentially before comingled placement on fresh pine shavings. Turkeys were challenged with 108 cfu SR by oral gavage on d 4 and 7 postplacement. Subsets were subjected to simulated commercial processing on d 14 (n = 40), 21 (n = 40) and 28 (n = 32) postplacement (corresponding to 10, 11, and 12 wk of age). Stifle joint, skin, trachea, crop, lung, liver + spleen (LS), and ceca were aseptically sampled and cultured for Salmonella recovery and serotyping. SR could not be recovered from stifle joint 14 d post inoculation (PI). However, at 14 d PI, recovery of SR were: Skin 80%; crop 75%; LS 67.5%; lungs 60%; and ceca 57.5%. (P < 0.01). Interestingly, the lowest recovery of SR was observed from trachea (40%). At 21 d PI, the highest rate of positive samples to SR were observed in ceca (87.5%) and crop (67.5%). By 28 d PI, SR was only recovered from ceca (75%); crop (43.8%); lung (34.4%); and LS (21.9%). The results of this study confirms that SR is an emerging problem for the turkey industry and immediate measurements to reduce foodborne pathogens such as Salmonella should target all parts of the supply chain and consumer education about food safety.