Ileal and cecal microbiota response to Salmonella Typhimurium challenge in conventional and slow-growing broilers.

Despite the negative impacts of Salmonella intestinal colonization on human health, Salmonella is a natural colonizer of the gastrointestinal tract and is not overtly pathogenic to the avian host. It is of interest to understand the impacts and colonization rates of Salmonella across selected genetic lines such as slow-growing (SG) and conventional (CONV) broilers. The objective of this study was to characterize the relationship between Salmonella enterica serovar Typhimurium challenge and selected broiler genetic lines on the ileal and cecal microbiome. Male chicks of two broiler breeds (n = 156/breed) were cohoused in an open floor pen until day 7. On day 13, the chicks were then separated into 12 isolators per breed (4 rooms, 6 isolators/room, 11 chicks/isolator). On day 14, chicks in the 12 treatment isolators (6 isolators/breed, 108 total) were challenged with Salmonella Typhimurium (ST) (1 × 10 8 CFU/ml) via oral gavage while the remaining chicks (n = 108) were given an oral gavage of sterile tryptic soy broth control (C). Ileal and cecal contents were collected on day 7 from 24 chicks of each breed, and on days 13, 17, 21, and 24 from two chicks per isolator. Samples underwent DNA extraction and PCR amplification to obtain 16S rRNA amplicons that were sequenced with Illumina MiSeq. Salmonella Typhimurium colonization in the cecum was not different in the two broiler breeds. The main effect of breed had the greatest impact on the ileum microbiota of broilers 7 days of age where SG broilers had significantly lower diversity and richness compared to CONV broilers (p < 0.05). Salmonella Typhimurium challenge consistently caused a change in beta diversity. Regardless of day or intestinal location, challenged broilers had many amplicon sequence variants (ASVs) with decreased abundance of likely beneficial bacteria such as Mollicutes RF39, Shuttleworthia, Flavonifractor, and Oscillibacter compared to broilers that were unchallenged with Salmonella Typhimurium (p < 0.05). Additionally, there was a difference in the timing of when the microbiota alpha and beta diversity of each breed responded to Salmonella Typhimurium challenge. Thus, both broiler breed and Salmonella Typhimurium can impact the intestinal microbiota.

Behavior and Immune Response of Conventional and Slow-Growing Broilers to Salmonella Typhimurium.

Fast growth rate in broiler chickens comes with welfare concerns and the contribution of growth rate to pathogen resistance and sickness behavior is relatively unknown. The objective of this study was to evaluate physiological and behavioral responses of conventional (CONV) and slow-growing (SG) male broilers challenged with Salmonella Typhimurium. CONV (n = 156) and SG (n = 156) chicks were raised in a pen with wood litter shavings until day 7 of age, when birds were transferred to 24 isolators (n = 11 chicks/isolator). On day 14 of age, half of the birds (n = 12 isolators) were challenged with S. Typhimurium (ST) and the other half (n = 12 isolators) received a control (C). On days 7, 13, 17, 21, and 24, body weight was recorded, and blood, jejunum and ileum sections were collected from 2 birds/isolator (n = 48 birds/sampling) to measure plasma IgA and IgG and intestinal histomorphology, respectively. On days 12, 16, 21, and 23, video was recorded to evaluate bird postures (sitting, standing, or locomoting) and behaviors (eating, drinking, preening, stretching, sham foraging, allopreening, and aggression). CONV birds were 70 g heavier (p = 0.03) on day 21 and 140 g heavier (p = 0.007) on day 24 than SG. On day 7, CONV jejunum villus height and crypt depth were 22 and 7 µm greater (p ≤ 0.001), respectively, than SG. On day 24, ST ileum villus height was 95 µm shorter (p = 0.009) than C. IgA increased after day 17 for all birds and at day 21, CONV IgA was greater (p = 0.01) than SG. Although SG IgG was 344 µg/ml greater (p = 0.05) than CONV on day 7, CONV IgG increased with age (p < 0.0001) to greater (p ≤ 0.03) concentrations than SG on day 21 and day 24 by 689 µg/ml and 1,474 µg/ml, respectively, while SG IgG remained at similar concentrations after day 13. Generally, a greater proportion of birds sham foraged as they aged (p < 0.0001). A greater proportion of CONV tended to sit (p = 0.09) and fewer locomoted (p < 0.0001) than SG as they aged. The results illustrate conventional and slow-growing broilers differ in their behavior, immunity, and response to Salmonella.

Impact of experimentally induced bacterial chondronecrosis with osteomyelitis (BCO) lameness on health, stress, and leg health parameters in broilers.

Stress and lameness negatively affect the health, production, and welfare of broilers. Bacterial chondronecrosis with osteomyelitis (BCO) is a leading cause of stress and lameness in commercial broilers. External changes in skin temperature related to changes in blood flow can be detected with infrared thermography (IRT), offering a noninvasive tool to assess the health of animals. This study compared physiological and noninvasive measures of stress and lameness in clinically healthy and lame male broiler chickens between 25 and 56 d. Birds were raised in pens within separate environmental chambers containing either litter flooring (sound) or wire flooring, with the latter established to induce BCO lameness (lame). Physiological and noninvasive measures of stress and lameness were collected: body weight, (BW), relative bursa weight, core body temperature, corticosterone (CORT) concentrations in serum and feathers, surface temperatures of the head (eye and beak) and leg (hock, shank, and foot) regions by infrared thermography (IRT), leg blood oxygen saturation (leg O2), and BCO lesion severity scores of tibial head necrosis (THN) and femoral head necrosis (FHN). Lame birds exhibited greater FHN and THN lesion severities, core body temperatures, and serum CORT (P < 0.05), but had lower BW, relative bursa weight, leg O2, and IRT surface temperatures of the beak, hock, shank, and foot compared with sound birds (P < 0.05). The difference in THN lesion severity between sound and lame birds decreased with age. Linear relationships between leg O2 with IRT leg surface temperatures were positive and negative between leg O2 with BCO lesion severity (P < 0.05). There were negative correlations between serum CORT with hock, shank and foot temperatures (P < 0.001), indicating that BCO is stressful. These results indicate that birds lame from BCO are stressed, have reduced oxygen saturation of blood in their legs, and that IRT surface temperatures can be used as noninvasive indicators of stress and lameness in broilers.

Broiler stress responses to light intensity, flooring type, and leg weakness as assessed by heterophil-to-lymphocyte ratios, serum corticosterone, infrared thermography, and latency to lie.

Stress and leg weakness are detrimental to broiler production, health, and welfare. Traditional methods to evaluate stress may be stressful to the bird because they are invasive and require handling and restraint. Two studies examined the effects of light intensity and flooring on the following in broilers: 1) traditional methods for assessing stress using heterophil-to-lymphocyte ratios and serum corticosterone (CORT) concentrations, 2) noninvasive measures of stress from infrared thermography (IRT) eye and beak surface temperatures, and 3) latency-to-lie (LTL) test times of birds tested individually and in groups of 5. Day-of-hatch male broiler chicks were placed into 6 pens (N = 120 chicks/pen). At 1 wk, pens were allocated to 3 light intensity treatments (2, 5, or 10 lux). At 4 wk, half of the birds from each pen were moved to a pen with wire flooring and the same light intensity. At 1, 4, 5, and 8 wk, blood samples were collected and IRT images of the heads of 5 clinically healthy broilers from each pen were captured. In study 2, IRT images of the heads of birds that became lame in the wire flooring pens were taken. There were no treatment effects on the LTL times of birds tested in groups or individually (P > 0.05). On day 56 in study 1, birds on wire flooring had elevated heterophil-to-lymphocyte ratios and CORT concentrations (P ≤ 0.002) and depressed IRT eye and beak temperatures (P < 0.0001). In both studies, there were negative correlations between CORT concentrations and IRT beak surface temperatures (P < 0.05). Lame birds had lower IRT eye and beak surface temperatures than sound birds (P ≤ 0.004), and the IRT beak surface temperatures of lame birds were lower than their eye surface temperatures (P = 0.004) in study 2. These studies indicate that the IRT surface temperatures of the eye, and more distinctly of the beak, can be used as sensitive noninvasive indicators of stress.

The utility of infrared thermography for evaluating lameness attributable to bacterial chondronecrosis with osteomyelitis.

Bacterial chondronecrosis with osteomyelitis (BCO) is a leading cause of lameness in broilers. Infrared thermography (IRT) is a noninvasive technique for measuring infrared radiation from an object and can be used to evaluate clinical health. Two replicated studies compared the effect of light intensity on broilers grown on a wire flooring model that experimentally increased their susceptibility to and incidence of BCO lameness. Day-of-hatch male broiler chickens were placed into 6 pens on wood shavings litter, and at 1 wk one of 3 light intensity treatments (2, 5, or 10 lux) was allotted. At 4 wk half of the population from each pen was moved to a pen with wire flooring and the same light intensity. At 1, 4, 5, and 8 wk, an IRT image of the legs of 5 clinically healthy broilers from each pen was taken. The right and left proximal femora and tibiae of sound and lame broilers were scored for femoral head necrosis (FHN) and tibial head necrosis (THN) lesion severity. There were minimal effects of light intensity and flooring. In Study 1, but not Study 2, broilers on wire flooring weighed less on day 38 (P = 0.007) and days 57 to 58 (P = 0.003) compared to those on litter. The proportion of broilers that became lame on wire flooring was 52% in Study 1 and 14% in Study 2. The proportion of sound broilers from litter and wire flooring pens with subclinical signs of BCO in their right or left proximal growth plates was over 45% for FHN and 92% for THN, and lame broilers had more severe (P < 0.0001) FHN and THN compared to sound broilers. IRT surface temperatures of the hock joint, shank, and foot were consistently lower (P < 0.0001) in broilers that became lame when compared to sound. Therefore, IRT surface temperatures of broiler leg regions may be useful for detecting lesions attributed to BCO.

An evaluation of methods for measuring stress in broiler chickens.

There are potential advantages for using noninvasive methods instead of conventional approaches for measuring corticosterone (CORT) as a metric of stress. Two studies compared blood heterophil: lymphocyte (H:L) ratios, serum CORT, cecal content CORT (Study 1), colon content CORT (Study 2), and feather CORT concentrations for broiler chicks receiving drinking water with or without added CORT. On day 28, male broilers (N = 140) were provided tap water (Control group) or water containing 20 mg/L of CORT (CORT group) for up to 72 h. Body weight (BW), blood, feather, cecal, and colon content samples were collected from 10 birds from each treatment group at 0, 6, 12, 24, 30, 48, and 72 h. Data were analyzed with a JMP Pro ANOVA. Mean comparisons were analyzed on significant treatment effects using post hoc t-tests to compare control and CORT measures within each sampling time point. Pearson's pairwise correlations for all data were performed. A treatment by time interaction affected all measures in both studies. In both studies, CORT treatment reduced body weight and increased H:L ratios at later time points. In study 1 cecal content CORT concentrations were elevated with CORT treatment after 24 h, whereas in study 2 colon content CORT concentrations were elevated after 6 h. CORT treatment increased body and primary feather CORT and primary feather CORT increased with time in control groups in both studies. Serum CORT exhibited the lowest variability compared with all other sample sources used for determining CORT. Estimates of CORT by different methods were positively correlated. These results indicate that serum CORT concentrations most reliably reflect the CORT status of broilers, and that feather CORT concentrations demonstrated potential for estimating stress in broilers.