Effect of trans-cinnamaldehyde nanoemulsion wash on chicken embryo development in fertilized eggs.

Cleanliness of eggs is critical in successful hatching egg operations. The objective of this study was to investigate the effect of trans-cinnamaldehyde nanoemulsion (TCNE) wash treatments, as a sanitation strategy, on embryonic development in fertilized eggs. Trans-cinnamaldehyde is a generally recognized as safe status phytochemical obtained from cinnamon bark. TCNE were prepared with emulsifiers Tween 80 (Tw.80) or gum Arabic and lecithin (GAL) by sonication. Day-old fertilized eggs were subjected to TCNE wash treatments at 34°C for 5 min, followed by 18 d of incubation at 37.7°C. Washing of fertilized eggs with TCNE-Tw.80 or GAL at 0.48% concentration did not significantly alter the egg weight at d 18 of incubation, as compared to baseline and control (P > 0.05). The egg weight loss (calculated as percentage) did not differ significantly between eggs subjected to nanoemulsion wash treatments and control eggs (P > 0.05). In case of embryo fertility and mortality, for baseline and control, ∼ 95% fertility rate was achieved, with combined early and midterm mortality at 16%. Likewise, TCNE-Tw.80 or TCNE-GAL resulted in 95% fertility (P > 0.05), with 11% and 17% combined early and midterm mortality, respectively. Furthermore, TCNE wash treatments did not differ significantly in yolk sac and embryo weight (as compared to control) and did not affect the length of the d 18 embryo (P > 0.05). Moreover, TCNE wash treatments did not alter tibia weight and length (P > 0.05). Results suggest that TCNE could potentially be used as a natural antimicrobial for fertilized egg sanitation. Further studies in industry settings are warranted.

Trans-cinnamaldehyde nanoemulsion wash inactivates Salmonella Enteritidis on shelled eggs without affecting egg color.

Salmonella Enteritidis is a major foodborne pathogen that causes enteric illnesses in humans, primarily through the consumption of contaminated poultry meat and eggs. Despite implementation of traditional disinfection approaches to reduce S. Enteritidis contamination, egg-borne outbreaks continue to occur, raising public health concerns and adversely affecting the popularity and profitability for the poultry industry. Generally Recognized as Safe (GRAS) status phytochemicals such as Trans-cinnamaldehyde (TC) have previously shown to exhibit anti-Salmonella efficacy, however, the low solubility of TC is a major hurdle in its adoption as an egg wash treatment. Therefore, the present study investigated the efficacy of Trans-cinnamaldehyde nanoemulsions (TCNE) prepared with emulsifiers Tween 80 (Tw.80) or Gum Arabic and lecithin (GAL) as dip treatments, at 34°C, for reducing S. Enteritidis on shelled eggs in presence or absence of 5% chicken litter. In addition, the efficacy of TCNE dip treatments in reducing trans-shell migration of S. Enteritidis across shell barrier was investigated. The effect of wash treatments on shell color were evaluated on d 0, 1, 7, and 14 of refrigerated storage. TCNE-Tw.80 or GAL treatments (0.06, 0.12, 0.24, 0.48%) were effective in inactivating S. Enteritidis by at least 2 to 2.5 log cfu/egg as early as 1 min of washing time (P < 0.05). In presence of organic matter, nanoemulsions (0.48%) reduced S. Enteritidis counts by ∼ 2 to 2.5 log cfu/egg as early as 1 min, (P < 0.05). Nanoemulsion wash also inhibited trans-shell migration of S. Enteritidis, as compared to control (P < 0.05). The nanoemulsion wash treatments did not affect shell color (P > 0.05). Results suggest that TCNE could potentially be used as an antimicrobial wash to reduce S. Enteritidis on shelled eggs, although further studies investigating the effect of TCNE wash treatments on organoleptic properties of eggs are necessary.

Carvacrol attenuates Campylobacter jejuni colonization factors and proteome critical for persistence in the chicken gut.

Campylobacter jejuni is a major foodborne pathogen that causes gastroenteritis in humans. Chickens act as the reservoir host for C. jejuni, wherein the pathogen asymptomatically colonizes the ceca leading to contamination of carcasses during slaughter. The major colonization factors in C. jejuni include motility, intestinal epithelial attachment, acid/bile tolerance, and quorum sensing. Reducing the expression of the aforementioned factors could potentially reduce C. jejuni colonization in chickens. This study investigated the efficacy of subinhibitory concentration (SIC; compound concentration not inhibiting bacterial growth) of carvacrol in reducing the expression of C. jejuni colonization factors in vitro. Moreover, the effect of carvacrol on the expression of C. jejuni proteome was investigated using liquid chromatography-tandem mass spectrometry. The motility assay was conducted at 42°C, and the motility zone was measured after 24 h of incubation. For the adhesion assay, monolayers of primary chicken enterocytes (∼105 cells/well) were inoculated with C. jejuni (6 log cfu/well) either in the presence or absence of carvacrol, and the adhered C. jejuni were enumerated after 90 min of incubation at 42°C. The effect of carvacrol on C. jejuni quorum sensing and susceptibility to acid/bile stress was investigated using a bioluminescence assay and an acid-bile survival assay, respectively. The SIC (0.002%) of carvacrol reduced the motility of C. jejuni strains S-8 and NCTC 81-176 by ∼50 and 35%, respectively (P < 0.05). Carvacrol inhibited C. jejuni S-8 and NCTC 81-176 adhesion to chicken enterocytes by ∼0.8 and 1.5 log cfu/mL, respectively (P < 0.05). Moreover, carvacrol reduced autoinducer-2 activity and increased the susceptibility of C. jejuni to acid and bile in both the strains (P < 0.05). Liquid chromatography-tandem mass spectrometry revealed that the SIC of carvacrol reduced the expression of selected C. jejuni colonization proteins critical for motility (methyl-accepting chemotaxis protein), adhesion (GroL), growth and metabolism (AspA, AcnB, Icd, Fba, Ppa, AnsA, Ldh, Eno, PurB-1), and anaerobic respiration (NapB, HydB, SdhA, NrfA) (P < 0.05). Results suggest the mechanisms by which carvacrol could reduce C. jejuni colonization in chickens.

Carvacrol antimicrobial wash treatments reduce Campylobacter jejuni and aerobic bacteria on broiler chicken skin.

Campylobacter jejuni, a major cause of human gastroenteritis worldwide, is often associated with the consumption of contaminated poultry products. With increasing consumer preference to natural and minimally processed foods, interventions utilizing natural antimicrobials for controlling C. jejuni on poultry products are gaining popularity. This study investigated the efficacy of the generally recognized as safe compound carvacrol (CR) as a wash treatment in reducing C. jejuni and aerobic bacteria on chicken skin. Two separate studies, each with 2 trials, were conducted. In the first study, the efficacy of CR suspension (0, 0.25, 0.5, 1, and 2%) was investigated, whereas in the second the efficacy of CR as suspension, emulsion, and nanoemulsion was studied. In both studies, skin samples were inoculated with 50 µL (∼8 log10 cfu/sample) of a cocktail of 4 wild strains of C. jejuni. After 30 min of attachment, samples were washed with the respective treatments for 1 min, drip dried for 2 min, and processed at 0, 8, 24, h post-treatment for enumeration of C. jejuni and aerobic bacterial counts (n = 5/treatment/time point). In addition, the effect of treatments on the color of chicken skin was evaluated. Data were analyzed using PROC MIXED procedure of SAS. In the first study, all the tested doses of CR suspension consistently reduced C. jejuni counts across all time points. The 2% CR suspension wash reduced C. jejuni counts by ∼2.4 to 4 log10 cfu/sample (P < 0.05). In addition, 1% and 2% CR suspensions significantly reduced aerobic counts at all the time points. The results from the second study suggest that anti-Campylobacter efficacy of CR emulsion or nanoemulsion treatments was not improved compared to CR suspension. Several CR suspension treatments were more effective than corresponding emulsion or nanoemulsion treatments. No significant differences were observed in the color of the samples between treatments (P > 0.05). The results suggest that CR could potentially be used as an antimicrobial wash treatment in postharvest poultry.

Pectin or chitosan coating fortified with eugenol reduces Campylobacter jejuni on chicken wingettes and modulates expression of critical survival genes.

Campylobacter jejuni infection in humans is strongly associated with the consumption of contaminated poultry products. With increasing consumer demand for minimally processed and natural product, there is a need for novel intervention strategies for controlling C. jejuni. Antimicrobial coatings are increasingly being used for preventing food contamination due to their efficacy and continuous protection of product. This study investigated the efficacy of pectin and chitosan coating fortified with eugenol to reduce C. jejuni on chicken wingettes. Pectin, chitosan, and eugenol are generally recognized as safe status compounds derived from berries, crustaceans, and cloves respectively. Each wingette was inoculated with a mixture of 4 wild-type strains of C. jejuni (approximately 107 CFU/sample) and randomly assigned to controls, pectin (3%), chitosan (2%), eugenol (0.5, 1, or 2%), or their combinations. Following 1 min of coating, wingettes were air-dried, vacuum sealed, and sampled on 0, 1, 3, 5, and 7 d of refrigerated storage for C. jejuni and aerobic counts (n = 5 wingettes/treatment/d). In addition, the effect of treatments on wingette color and expression of C. jejuni survival/virulence genes was evaluated. All 3 doses of eugenol or chitosan significantly reduced C. jejuni and aerobic bacteria from 0 d through 7 d. Incorporation of 2% eugenol in chitosan improved coating efficiency and reduced C. jejuni counts by approximately 3 Log CFU/sample at the end of 7 d of storage (P < 0.05). Similarly, the antimicrobial efficacy of pectin was improved by 2% eugenol and the coating reduced C. jejuni by approximately 2 Log CFU/sample at 7 d of storage. Chitosan coating with 2% eugenol also showed greater reductions of total aerobic counts as compared to individual treatments of eugenol and chitosan. No significant difference in the color of chicken wingettes was observed between treatments. Exposure of C. jejuni to eugenol, chitosan, or combination significantly modulated select genes encoding for motility, quorum sensing, and stress response. Results demonstrate the potential of pectin or chitosan coating fortified with eugenol as a postharvest intervention against C. jejuni contamination on poultry products.

ß-Resorcylic Acid, a Phytophenolic Compound, Reduces Campylobacter jejuni in Postharvest Poultry.

Human Campylobacter infections, a leading foodborne illness globally, has been linked with the high prevalence of this bacterium on raw retail chicken products. Reduction of Campylobacter counts on poultry products would greatly reduce the risk of subsequent infections in humans. To this end, this study investigated the potential of the phytophenolic compound ß-resorcylic acid (BR) to reduce Campylobacter counts on postharvest poultry (chicken skin or meat). Four trials in total, two each on thigh skin or breast meat, were conducted in which chicken skin or meat samples (2 ± 0.1 g; 10 samples per treatment) were inoculated with 50 µL (∼106 CFU per sample) of a cocktail of four wild strains of C. jejuni. After 30 min of attachment, inoculated samples were dipped in a 0, 0.5, 1, or 2% BR solution for 30 s immediately followed by vigorously vortexing the samples in Butterfield's phosphate diluent and plating the supernatant for Campylobacter enumeration. In addition, the effect of BR on the color of skin and meat samples was studied. Moreover, the change in the expression of survival and virulence genes of C. jejuni exposed to BR was evaluated. Data were analyzed by the PROC MIXED procedure of SAS (P < 0.05; SAS Institute Inc., Cary, NC). All BR treatments significantly reduced Campylobacter populations on both chicken or meat samples by 1 to 3 log CFU/g compared with non-BR-treated washed controls. No significant difference in the lightness, redness, and yellowness of skin and meat samples was observed on exposure to BR wash (P > 0.05). Real-time PCR results revealed that BR treatment down-regulated expression of select genes coding for motility (motA, motB) and attachment (cadF, ciaB) in the majority of C. jejuni strains. Stress response genes (sodB, katA) were upregulated in C. jejuni S-8 (P < 0.05). Overall, our results suggest that BR could be effectively used as antimicrobial dip treatment during poultry processing for reducing Campylobacter on chicken carcasses.

Nutritional Supplement of Hatchery Eggshell Membrane Improves Poultry Performance and Provides Resistance against Endotoxin Stress.

Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM) as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS) challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase resistance of chickens to endotoxin.

Effect of outdoor structural enrichments on the performance, use of range area, and behavior of organic meat chickens.

Chickens provided with outdoor access often do not fully use the range area. Natural cover, such as trees, can provide shelter and increase range use, but may not be practical for use in all free-range operations. A study was conducted to determine whether constructed enrichments that simulate natural structures increase range use and impact behavior of meat chickens. Slow-growing Delaware chickens were raised in floor pens (17 birds/pen) in a naturally ventilated house, with a pophole in each pen that allowed daily access to a vegetation-covered outdoor area. Feed and water were provided inside and outside. Birds were randomly assigned to one of 2 treatments: no range enrichment (control; NON) or range enrichment (ENR), which had roosts made of plastic pipe or screened shelters placed 7.6 m and 22.9 m from the house and overhead shade panels at 15.2 m. There were 4 replications. The numbers of birds inside and in the 4 quadrants of the range were counted in 7-min intervals 3 times daily (0900 h, 1300 h, and 1600 h) for 3 consecutive d when the birds were 7 and 10 wk of age; the numbers of birds performing various behaviors also were counted. On average, only 12.9% of birds used the range at any given time. The behavior of the birds was strongly affected by location (inside vs. outside), with birds mainly foraging and walking while outside and feeding, standing, and sitting while inside. Time of d impacted the number of birds that ranged (P ≤ 0.0001), with fewest birds outside during mid-day. Overall, birds using the range were most often (71%) observed in the quadrant nearest the house (zero to 7.6 m). However, in the ENR treatment, more birds (13%; P ≤ 0.0001) were observed in the farthest 2 quadrants (15.2 to 30.5 m from the house) than in the NON treatment (6%). This indicates that adding constructed enrichments to the range encouraged meat chickens to use it more fully.

Hearing conservation in the primary aluminium industry.

BACKGROUND: Noise-induced hearing loss has been an intractable problem for heavy industry. AIMS: To report our experience in reducing the incidence of age-corrected confirmed 10 dB hearing shifts (averaged over 2, 3 and 4 kHz) in employees in the primary aluminium industry in Australia over the period 2006-13. METHODS: We analysed annual audiometric data to determine the number of permanent hearing shifts that occurred in employees in two bauxite mines, three alumina refineries and two aluminium smelters. Annual hearing shift rates were calculated based on the number of employees tested per year. Hearing conservation initiatives undertaken during the study period are described. An assessment of similar exposure group noise exposures was also undertaken to determine the magnitude of noise exposure reduction during the study period. RESULTS: Across all operations, hearing shift rates declined from 5.5% per year in 2006 to 1.3% per year in 2013 (P < 0.001). The decline in shift rates was greater in mines and refineries, where baseline shift rates were higher, than in smelter workers. Modest reductions in noise exposure occurred during the study period. CONCLUSIONS: We observed a substantial decline in hearing shift rates during the study period. We describe the hearing conservation initiatives that were collectively associated with this decline. We suspect these initiatives could be deployed relatively easily and at modest cost in other industries with noise-exposed employees.

Intracloacal inoculation, an effective screening method for determining the efficacy of probiotic bacterial isolates against Campylobacter colonization in broiler chickens.

Campylobacter is a leading cause of foodborne illness worldwide. It is common in poultry, and human infections are often associated with consumption of contaminated poultry products. One strategy to reduce Campylobacter colonization in poultry is the use of oral probiotics, but this produces variable results, possibly because the probiotics are destroyed in the stomach's acidic environment. Protection (e.g., encapsulation) of isolates may overcome this problem, but there is no assurance that these isolates will have efficacy in the lower gastrointestinal tract. Therefore, screening candidate isolates by directly placing them in the lower intestinal tract via cloacal inoculation may eliminate the time and expense of encapsulating ineffective isolates. Thus, the purpose of this study was to collect bacterial isolates with anti-Campylobacter activity in vitro and evaluate their efficacy in vivo upon either oral or intracloacal administration. Bacterial isolates were collected from healthy birds and were evaluated for efficacy against C. jejuni in vitro. Isolates having generally regarded as safe status and demonstrating in vitro anti-Campylobacter properties were evaluated after oral or intracloacal inoculation into chicks on day 1 (n = 10 birds per isolate per route of administration). On day 7, birds were dosed by oral gavage with a four-strain mixture of wild-type Campylobacter containing at least 1 × 10(7) CFU/ml organisms. On day 14, birds were euthanized and the ceca were collected aseptically for Campylobacter enumeration. When dosed orally, only one isolate had a 1-log reduction in cecal Campylobacter counts, whereas when administered intracloacally, six of these isolates produced a 1- to 3-log reduction in cecal Campylobacter counts in 14-day-old chickens. These results support the strategy of evaluating the efficacy of potential probiotic isolates via cloacal inoculation prior to undergoing the effort of encapsulating isolates for oral administration.

Environmental augmentation with bacteriophage prevents colibacillosis in broiler chickens.

Bacteriophages are viruses that kill bacteria. They are plentiful in nature; are safe, having no known activity to human or animal cells; and are an attractive alternative to antibiotics. The objectives of this research were to establish an experimental model of colibacillosis induced by indirect exposure to Escherichia coli and to determine if bacteriophage could protect the birds from developing colibacillosis. In study 1 there were 6 treatments with 2 replicate pens of 25 birds. The treatments were control warm brooded; control cold stressed; litter inoculated with E. coli, warm brooded; litter inoculated with E. coli, cold stressed; seeder birds (5 per pen) challenged with E. coli, warm brooded; and seeder birds (5 per pen), cold stressed. The study concluded when the birds were 3 wk of age. Body weights at 1, 2, and 3 wk of age were significantly decreased (P ≤ 0.05) by cold stress, decreased at 1 and 2 wk of age by both the litter and seeder bird treatments compared with the control treatment and by the seeder bird treatment at 3 wk of age. Study 2 consisted of 8 treatments with 2 replicate pens of 20 birds per treatment. The treatments were control, warm brooded; control, cold stressed; litter inoculated with E. coli, cold stressed; and seeder birds (5/pen) challenged with E. coli, cold stressed with and without bacteriophage treatment. In the bacteriophage treatments the bacteriophages were sprayed on the litter. The study was concluded at 3 wk of age. Body weights at 1 wk of age were significantly (P ≤ 0.05) decreased from the control treatment by the seeder bird treatment and were significantly (P ≤ 0.05) higher in all the bacteriophage treatments compared with their matched untreated treatments, except in the control cold stressed treatment. Mortality was significantly (P ≤ 0.05) decreased by bacteriophage in the litter challenged treatment. These data suggest that augmentation of the environment with bacteriophage is a practical and efficacious way to prevent colibacillosis in broiler chickens.

Method of administration affects the ability of bacteriophage to prevent colibacillosis in 1-day-old broiler chickens.

Bacteriophages are viruses that kill bacteria. They are plentiful in nature with no known activity in human or animal cells, making them an attractive alternative to antibiotics. The objective of this research was to determine if a coarse or a fine spray of bacteriophage would prevent colibacillosis induced by an intratracheal (IT) challenge with Escherichia coli. Two studies were conducted with 6 treatments: untreated control, birds treated with a spray administration of bacteriophage and not challenged, birds administered bacteriophage IT and not challenged, birds not treated and challenged IT with E. coli, birds sprayed with bacteriophage and IT challenged with E. coli, and birds administered bacteriophage IT and challenged IT with E. coli. There were 3 replicate pens of 10 birds per pen, per treatment, and all treatments were administered at 1 d of age. Study 1 was concluded when the birds were 19 d of age, and study 2 was concluded when the birds were 21 d of age. In both studies, neither a coarse nor a fine spray protected the birds from an IT E. coli challenge; however, when bacteriophage was administered IT there was complete protection. This research demonstrates the necessity for the administration of bacteriophage therapeutics to deliver high bacteriophage titers to the site of a bacterial infection.

Use of plant-derived antimicrobials for improving the safety of poultry products.

Salmonella Enteritidis and Campylobacter jejuni are the 2 major foodborne pathogens transmitted through poultry products. Chickens are the reservoir hosts of these pathogens, with their intestinal colonization being the most significant factor causing contamination of meat and eggs. Effective preslaughter strategies for reducing the colonization of birds with these pathogens are critical to improve the microbiological safety of poultry products. An antimicrobial treatment that can be applied through feed represents the most practical and economically viable method for adoption on farms. Additionally, a natural and safe antimicrobial will be better accepted by producers without concerns for toxicity. This symposium talk discussed the potential use of plant-derived, GRAS (generally recognized as safe)-status molecules, caprylic acid, trans-cinnamaldehyde, eugenol, carvacrol, and thymol as feed supplements for reducing cecal populations of Salmonella Enteritidis and C. jejuni in chickens. Additionally, the effect of plant molecules on Salmonella virulence genes critical for cecal colonization in chickens was also discussed.

Caprylic acid reduces Salmonella Enteritidis populations in various segments of digestive tract and internal organs of 3- and 6-week-old broiler chickens, therapeutically.

We investigated the efficacy of feed supplemented with caprylic acid (CA), a natural, 8-carbon fatty acid for reducing Salmonella enterica serovar Enteritidis colonization in commercial broiler chickens. In separate 3- and 6-wk trials, 1-d-old straight-run broiler chicks (n = 70 birds/trial) were assigned to a control group (challenged with Salmonella Enteritidis, no CA) and 2 replicates of 0.7 and 1% CA (n = 14 birds/group). Water and feed were provided ad libitum. On d 1, birds were tested for any inherent Salmonella (n = 2 birds/group). For the 3-wk trial, on d 5, birds were challenged with 8 log(10) cfu of Salmonella Enteritidis of a 4-strain mixture by crop gavage, and after 5 d postchallenge, birds (n = 2 birds/group) were euthanized to ensure Salmonella Enteritidis colonization. Caprylic acid was supplemented the last 5 d before tissue collection (n = 10 birds/group). For the 6-wk trial, on d 25, birds were challenged and confirmed for Salmonella Enteritidis colonization. The birds (n = 10 birds/group) were euthanized for tissue samples after CA supplementation for the last 5 d. Caprylic acid at 0.7 or 1% decreased Salmonella Enteritidis populations in cecum, small intestine, cloaca, liver, and spleen in both 3- and 6-wk trials. Body weight of birds did not differ between the groups (P ≥ 0.05). Further, to elucidate a potential antibacterial mechanism of action of CA, we investigated if CA could reduce Salmonella Enteritidis invasion of an avian epithelial cell line and expression of invasion genes hilA and hilD. The cell invasion study revealed that CA reduced invasive abilities of all Salmonella Enteritidis strains by ~80% (P < 0.05). Gene expression studies indicated that CA downregulated (P < 0.001) Salmonella invasion genes hilA and hilD. These results suggest that supplementation of CA through feed could reduce Salmonella Enteritidis colonization in broiler chicken and potentially reduces the pathogen's ability to invade intestinal epithelial cells by downregulating key invasion genes, hilA and hilD.

Water administration of the medium-chain fatty acid caprylic acid produced variable efficacy against enteric Campylobacter colonization in broilers.

Campylobacter is one of the most common causes of foodborne illness, and poultry are considered a primary source of Campylobacter infections. Caprylic acid, an 8-carbon fatty acid, has been shown in previous studies to reduce enteric cecal Campylobacter concentrations in poultry when administered in the feed. For greater ease of application for producers, a water-soluble form of caprylic acid, sodium octanoate, was evaluated for efficacy against enteric Campylobacter. The first trial consisted of 70 birds in 7 groups (n = 10 chicks/group): an untreated control and 6 other groups that were challenged with Campylobacter at d 3 and that received 0, 0.175, 0.35, 0.7, 1.4, or 2.8% water-soluble caprylic acid in water 3 d before necropsy at d 14. The second trial consisted of 80 birds in 8 groups (n = 10 chicks/group): an untreated negative control and 7 other groups, all of which were challenged with Campylobacter at d 3 and received 0, 0.044, 0.088, 0.175, 0.35, 0.7, or 1.4% water-soluble caprylic acid for 3 d before necropsy at d 14. In trial 1, only the 0.175% dose caused a reduction in cecal Campylobacter counts in comparison with the positive control (approximately a 3-log reduction). In trial 2, no treatment reduced Campylobacter counts compared with the positive control. Unlike the efficacy of caprylic acid in feed, treatment with caprylic acid in water had an inconsistent effect on intestinal Campylobacter counts.

Immune interference of bacteriophage efficacy when treating colibacillosis in poultry.

A study was conducted to determine if prior exposure with bacteriophage would limit the ability of the same bacteriophage to treat colibacillosis. There were 5 treatments with 3 replicate pens of 20 birds per pen. The treatments consisted of 1) control, 2) birds treated with bacteriophage at 10 and 17 d of age, 3) birds challenged with Escherichia coli at 17 d of age, 4) birds challenged with E. coli and treated with bacteriophage at 17 d of age, and 5) birds treated with bacteriophage at 10 d of age and challenged with E. coli and treated with bacteriophage at 17 d of age. Colibacillosis was induced by injecting 0.1 mL of E. coli into the thoracic airsac containing 1 x 10(6) cfu. The bacteriophage was administered by i.m. injection of 0.1 mL into the thigh, providing a dose of 6.8 x 10(8) pfu. The study was concluded 14 d after E. coli challenge. Mortality in the birds challenged with E. coli and not treated with bacteriophage was 55% (treatment 3), and bacteriophage therapy significantly (P < or = 0.05) reduced mortality to 8% (treatment 4), which was not significantly (P < or = 0.05) different from the 2 non-E. coli-challenged controls (3%, treatment 1, and 2%, treatment 2). However, mortality in the birds administered bacteriophage before challenge with E. coli and treated with bacteriophage (treatment 5) was 33%, which was not significantly different (P < or = 0.05) from the birds that were challenged with E. coli and untreated (55%, treatment 3). A kinetic in vitro assay of bacteriophage activity found that serum from birds pretreated with bacteriophage (treatment 5) inhibited bacteriophage activity. The IgG levels to the bacteriophage in serum from birds pretreated with bacteriophage (treatment 2) were significantly higher at all dilutions compared with control serum (treatment 1). These data demonstrate that prior exposure to bacteriophage will limit bacteriophage therapeutic efficacy and suggests that the reduced efficacy is due to an immune response to bacteriophage.

Comparisons of sperm storage tubule distribution and number in 4 strains of mature broiler breeders and in turkey hens before and after the onset of photostimulation.

The biological basis of sustained fertility in broiler and turkey hens is their capacity to store sperm in the oviductal sperm storage tubules (SST) located in the uterovaginal junction. The objectives of this study were to determine if the numbers of SST varied between 4 strains of broiler breeders and determine the number of SST in the turkey before (less than 9 d of photostimulation) and after (up to 22 d of photostimulation and laying) photostimulation. No statistical differences were observed in SST numbers in the 4 strains of broilers examined or in turkey hens before and after the onset of egg production. The mean numbers of SST for broilers and turkeys were 4,893 and 30,566, respectively. We conclude that any differences between the fertility of the 4 broiler breeder strains examined cannot be explained by differences in SST numbers. However, differences in the duration of fertility between broilers and turkeys are, in part, related to their respective numbers of number of SST. Furthermore, we conclude that turkey SST are morphologically differentiated and functional before the onset of photostimulation and while the oviduct is morphologically undeveloped.

Bacterial clearance, heterophil function, and hematological parameters of transport-stressed turkey poults supplemented with dietary yeast extract.

Yeast extracts (YE) contain biological response modifiers that may be useful as alternatives to antibiotics for controlling pathogens in poultry production and mitigating the deleterious effects of production stressors. The objective of the present study was to determine the ability of a commercial dietary YE (Alphamune) to modulate the immune response in male turkey poults challenged with Escherichia coli and subjected to transport stress. Alphamune was added to turkey poult diets at 0, 500, or 1,000 g/ton. Poults were challenged by air sac injection with 60 cfu of E. coli at 1 wk of age. At 3 wk of age, these challenged birds were subjected to transport stress and birds were bled and necropsied the following morning. Blood cell numbers and percentages, hematological parameters, and clinical chemistry values were determined. Oxidative burst activity of isolated heterophils was measured using stimulation with phorbol myristate acetate and a 2',7'-dichlorofluorescein diacetate assay. Data were analyzed using GLM and least squares means procedures of the SAS program. The numbers and percentages of heterophils in peripheral blood were increased and their oxidative burst activity was stimulated by YE. The stress challenge dramatically increased oxidative burst and this increase was significantly modulated by YE treatment. Serum levels of calcium, phosphorus, and triglycerides were decreased and uric acid levels, erythrocyte numbers, hemoglobin, and hematocrit were increased by YE supplementation. Bacteria were isolated from the air sac and liver of a lower percentage of birds provided with YE. These results suggest that dietary YE has potential as a nonantibiotic alternative for decreasing bacterial pathogens in turkey production.

Implementing artificial insemination as an effective tool for ex situ conservation of endangered avian species.

Approximately 503 of the known species of birds are classified as 'endangered' or 'critical'. Captive propagation programs have proven useful in maintaining genetic diversity and restoring wild populations of certain species, including the Peregrine falcon, California condor and Whooping crane. Artificial insemination (AI) has the potential of solving problems inherent to reproductive management of small, closed populations of endangered birds, including dealing with demographic instability, physical and behavioral disabilities, sexual incompatibility, lack of synchrony, and need to maintain gene diversity. In this review, we address the necessary methods and factors that allow AI to be applied effectively to manage rare bird populations. It is clear that semen availability and quality are the greatest limiting factors to implementing consistently successful AI for birds. Behavioral sensitivity to animal handling and the ability to minimize stress in individual birds also are keys to success. Multiple, deep vaginal inseminations can improve fertility, particularly when semen quality is marginal. Laparoscopic methods of semen transfer also have produced fertile eggs. All of these practices leading to successful AI remain dependent on having adequate basic knowledge on female reproductive status, copulatory behavior, endocrine profiles and duration of fertility, especially as related to oviposition. The overall greatest challenge and highest priority is defining these normative traits, which are highly species-specific.