Differences in abilities to colonize reproductive organs and to contaminate eggs in intravaginally inoculated hens and in vitro adherences to vaginal explants between Salmonella enteritidis and other Salmonella serovars

In Experiment 1, mature laying hens were inoculated intravaginally with 10(6) colony-forming units of Salmonella enterica serovar enteritidis (S. enteritidis), Salmonella typhimurium, Salmonella infantis, Salmonella hadar, Salmonella heidelberg, or Salmonella montevideo to compare their abilities to colonize the reproductive organs of chickens and to contaminate eggs. Salmonella enteritidis was more frequently recovered (from 11 of 40 eggs, 27.5%) than the other serovars, and especially the inner shell was contaminated with these organisms in 10 of 40 eggs (25.0%). The contamination rates and the viable counts in cloaca were significantly (P < 0.05) higher in hens inoculated with S. enteritidis than in those inoculated with the other serovars at 4 days postinoculation (PI). In the vagina, the positive rates were 90%-100% in hens inoculated with S. enteritidis, and the viable counts of the organisms in this portion were significantly (P < 0.05) higher than those of the other serovars at 2, 4, and 7 days PI. The ceca were colonized similarly by each serovar at 7 days PI. The spleen and ovary were infected with S. enteritidis in three and one hen, respectively. No Salmonella was recovered from liver and peripheral blood in any hen. Salmonella enteritidis was recovered from other oviductal portions than the vagina (10%-20%), whereas no forming egg was contaminated in the oviduct. In Experiment 2, the in vitro adherence of these six serovars to the vaginal epithelium was compared with vaginal explants. The mean number of S. enteritidis attaching to the secondary villi in the vaginal lumen was significantly (P < 0.05) higher than those of the other serovars. These results suggest that S. enteritidis has a specific advantage over the other Salmonella serovars by its capacity to colonize the vaginal tissues of hens, and this higher affinity of S. enteritidis to the vagina may play a significant role in the production of many S. enteritidis-contaminated eggs.     

Colonization of reproductive organs and internal contamination of eggs after experimental infection of laying hens with Salmonella heidelberg and Salmonella enteritidis

Internal contamination of eggs laid by hens infected with Salmonella enteritidis has been a prominent international public health issue since the mid-1980s. Considerable resources have been committed to detecting and controlling S. enteritidis infections in commercial laying flocks. Recently, the Centers for Disease Control and Prevention also reported a significant association between eggs or egg-containing foods and S. heidelberg infections in humans. The present study sought to determine whether several S. heidelberg isolates obtained from egg-associated human disease outbreaks were able to colonize reproductive tissues and be deposited inside eggs laid by experimentally infected hens in a manner similar to the previously documented behavior of S. enteritidis. In two trials, groups of laying hens were orally inoculated with large doses of four S. heidelberg strains and an S. enteritidis strain that consistently caused egg contamination in previous studies. All five Salmonella strains (of both serotypes) colonized the intestinal tracts and invaded the livers, spleens, ovaries, and oviducts of inoculated hens, with no significant differences observed between the strains for any of these parameters. All four S. heidelberg strains were recovered from the interior liquid contents of eggs laid by infected hens, although at lower frequencies (between 1.1% and 4.5%) than the S. enteritidis strain (7.0%).     

Salmonella enteritidis and other Salmonella in laying hens and eggs from flocks with Salmonella in their environment.

Seven Canadian layer flocks with Salmonella enteritidis in their environment were investigated to determine the numbers of hens infected with S. enteritidis, the localization of S. enteritidis in organs of infected hens and the numbers of S. enteritidis-infected eggs produced by two affected flocks. By a microagglutination test (MAT) using S. pullorum antigens, these flocks had more seropositive hens (mean 51.9 +/- 16.9%) than two Salmonella-free flocks (mean 13.0 +/- 4.2%). Culture of tissues of 580 hens (433 seropositive) from the seven flocks detected 26 (4.5%) S. enteritidis-infected hens from two flocks. In one flock, 2/150 hens were infected with S. enteritidis phage type (PT) 8, which was confined to the ceca, and no Salmonella spp. were isolated from 2520 eggs (one day's lay). In the second flock, where 24/150 hens were infected with S. enteritidis PT13, extraintestinal infection was found in nine hens and involved the ovaries and/or oviduct in two hens. Salmonella enteritidis PT13 was isolated from one sample of egg contents and from one sample of cracked shells from among 14,040 eggs (one day's lay) from this flock. The overall prevalence of S. enteritidis-contaminated eggs from the two flocks with infected hens was less than 0.06%. Other Salmonella spp. isolated were S. heidelberg from 58 hens (10%), and S. hadar, S. mbandaka and S. typhimurium from one hen (0.2%) each. The MAT with antigens of S. pullorum had a sensitivity of 81% and a specificity of 24% for detecting S. enteritidis-infected hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Impacts of dried whole eggs on productive performance, quality of fresh and stored eggs, reproductive organs and lipid metabolism of laying hens

1. Rejected eggs were processed by freezing, freezing and boiling, or autoclaving, dried at 55°C, and the chemical and amino-acid analysis and microbiological properties of the dried whole eggs (DWE) determined. The processed DWE was included in a laying-hen mash diet at 50 or 100 g/kg, mainly to replace soybean meal and maize, in a factorial design of 3×2 and fed to hens between 25 and 41 weeks of age. In addition, a control diet without DWE was used to create a total of 7 treatments; each having 5 replicates of 5 hens. 2. Hens fed the diet containing 50-g/kg frozen and boiled DWE and 100-g/kg frozen DWE laid the largest egg size, while hens fed the 50-g/kg frozen and boiled or autoclaved DWE diets yielded the highest egg mass and best feed conversion efficiency. 3. Hens fed the 50-g/kg frozen DWE laid eggs with a higher yolk indices, Haugh unit scores, and eggshell thickness and maintained higher Haugh unit scores during egg storage. 4. Yolk cholesterol significantly decreased in eggs from groups fed either the 50-g/kg frozen or autoclaved DWE diet, with freezing having the stronger effect. 6. In conclusion, 50-g/kg autoclaved DWE diets fed to laying hens between 25 and 41 weeks of age had no adverse effects on production, egg quality or reproductive organs, but decreased yolk cholesterol, and thus could be beneficially included in laying-hen diets.     

Horizontal transmission of Salmonella and Campylobacter among caged and cage-free laying hens

In each of five sequential trials, laying hens (56-72 wk of age) were challenged with Salmonella and Campylobacter, and 1 wk postinoculation, the challenged hens (n = 3) were commingled with nonchallenged hens (n = 12) in conventional wire cages, on all-wire slats, or on all-shavings floor housing systems. After 12 days, challenged and nonchallenged hens were euthanatized for sample collection. Ceca were aseptically collected from all hens, and the spleen, liver/gallbladder (LGB), lower (LRT) and upper (URT) reproductive tracts, and ovarian follicles (mature and immature) were collected from only the challenged hens after commingling. Samples were divided equally and cultured separately for Salmonella and Campylobacter. Differences in the horizontal transmission of the challenge Salmonella to nonchallenged hens housed in cages (12%), on slats (15%), and on shavings (14%) were not significantly different (P > 0.05) from the challenged pen-mate hens over the five trials. However, with the inclusion of residual environmental Salmonella, the recovery of Salmonella from nonchallenged hens housed in cages was lowest at 15%, intermediate for hens on slats at 20%, and highest for hens on shavings at 38%. Among challenged hens housed in cages, Salmonella was recovered from only 27% of the cecum and LRT samples. From challenged hens housed on slats, Salmonella was recovered from 38% of the cecum, 12% of the spleen, 19% of the LGB, 44% of the LRT, and 19% of the URT samples. From challenged hens housed on shavings, Salmonella was recovered from 31% of the cecum; 15% of the spleen, LGB, and URT; and 31% of the LRT samples. Horizontal transmission of Campylobacter among nonchallenged pen-mate hens was significantly lower for hens housed in cages at 28% than for hens on shavings at 47%, with hens on slats being intermediate at 36%. For challenged hens housed in cages, Campylobacter was recovered from 27% of the cecum, 13% of the LRT, 7% of the URT, and 17% of the follicle samples. Among the challenged hens housed on slats, Campylobacter was recovered from 44% of the cecum, 6% of the spleen, 19% of the LGB, 12% of the LRT, 6% of the URT, and 14% of the follicle samples. Among challenged hens housed on shavings, Campylobacter was recovered from 46% of the cecum, 8% of the LRT and URT, and 40% of the follicle samples. The overall results of this study indicate that the caged housing system provided the lowest horizontal transmission level of Salmonella and Campylobacter among egg-laying hens.     

The relationship between the numbers of Salmonella Enteritidis, Salmonella Heidelberg, or Salmonella Hadar colonizing reproductive tissues of experimentally infected laying hens and deposition inside eggs

Contamination of eggs by Salmonella Enteritidis has been a prominent cause of human illness for several decades and is the focus of a recently implemented national regulatory plan for egg-producing flocks in the United States. Salmonella Heidelberg has also been identified as an egg-transmitted pathogen. The deposition of Salmonella strains inside eggs is a consequence of reproductive tract colonization in infected laying hens, but prior research has not determined the relationship between the numbers of Salmonella that colonize reproductive organs and the associated frequency of egg contamination. In the present study, groups of laying hens in two trials were experimentally infected with large oral doses of strains of Salmonella Enteritidis (phage type 13a), Salmonella Heidelberg, or Salmonella Hadar. Reproductive tissues of selected hens were cultured to detect and enumerate Salmonella at 5 days postinoculation, and the interior contents of eggs laid between 6 and 25 days postinoculation were tested for contamination. Significantly more internally contaminated eggs were laid by hens infected with Salmonella Enteritidis (3.58%) than with strains of either Salmonella Heidelberg (0.47%) or Salmonella Hadar (0%). However, no significant differences were observed between Salmonella strains in either isolation frequency or the number of colony-forming units (CFU) isolated from ovaries or oviducts. Salmonella isolation frequencies ranged from 20.8% to 41.7% for ovaries and from 8.3% to 33.3% for oviducts. Mean Salmonella colonization levels ranged from 0.10 to 0.51 log CFU/g for ovaries and from 0.25 to 0.46 log CFU/g for oviducts. Although parallel rank-orders were observed for Salmonella enumeration (in both ovaries and oviducts) and egg contamination frequency, a statistically significant relationship could not be established between these two parameters of infection.     

光照周期和饲喂水平对蛋鸡生殖器官发育的影响

本试验旨在研究限饲和光照周期对蛋鸡卵巢和输卵管发育及形态学的影响。试验采用3×2双因子设计,即3个光照周期（8L：16D、12L：12D和16L：8D）和2个饲喂水平（自由采食和限饲鸡自由采食量的80%）。选用体质健康的10周龄海兰灰蛋鸡240只,分成6组,每组4个重复,每个重复10只鸡。结果表明,光照周期显著影响20 w输卵管重（P〈0.05）,长光照周期（16L：8D）组和中长光照周期（12L：12D）组的20 w输卵管重显著高于短光照周期（8L：16D）组,对卵巢卵泡直径及单位面积内卵巢卵泡个数影响不显著（P〉0.05）;饲喂水平对20 w体重、卵巢重、输卵管重及卵巢卵泡直径和单位面积内卵巢卵泡个数影响均不显著（P〉0.05）;光照周期和饲喂水平互作对20 w卵巢卵泡直径影响显著（P〉0.05）,其中,β和VI组卵巢卵泡直径显著高于V组（P〈0.05）,其他各组间差异不显著（P〉0.05）。饲喂水平、光照周期影响输卵管发育,Ⅱ、Ⅳ、VI自由采食组发育程度优于Ⅰ、Ⅲ、V限饲组,并且16L：8D光照组输卵管的发育状况最好。     

Characteristics of Salmonella enteritidis contamination in eggs after oral,aerosol, and intravenous inoculation of laying hens

Experimental infection models are useful tools for understanding how Salmonella enteritidis is deposited in eggs and for testing potential strategies to control eggborne transmission of disease to humans. Oral inoculation of laying hens is presumed to provide the closest simulation of naturally occurring infections, but alternatives such as intravenous or aerosol inoculation have sometimes been recommended as options to induce higher incidences of egg contamination. The present study compared the frequency, level, and location of S. enteritidis deposition in egg contents after experimental inoculation by three different routes. In two replicate trials, specific-pathogen-free laying hens were infected with an S. enteritidis culture mixture prepared to optimize invasive behavior. Groups of hens received either an oral dose of 10(9) S. enteritidis, an aerosol dose of 10(9) S. enteritidis, or an intravenous dose of 10(5)-10(7) S. enteritidis. Oral inoculation led to the highest incidence of fecal shedding of S. enteritidis, whereas intravenous inoculation produced the highest specific antibody titers. Eggs laid during the first 21 days postinoculation were cultured to detect and enumerate S. enteritidis in the yolk and albumen. No significant differences were observed among the three inoculation routes in the frequencies of isolation of S. enteritidis from either yolk or albumen. For all three routes of administration, S. enteritidis was recovered more often from yolk (at frequencies ranging from 4% to 7%) than from albumen (0 to 2%). Over 73% of contaminated eggs harbored fewer than 1 colony-forming unit (CFU) of S. enteritidis per milliliter, and only 3% of such eggs contained more than 100 CFUs/ml. Significantly higher levels of S. enteritidis contaminants were associated with intravenous inoculation than with the other routes. No advantage of using aerosol or intravenous administration of S. enteritidis as an alternative to oral inoculation for inducing the production of contaminated eggs was evident in this study.     

Lead concentrations in the blood and eggs of backyard laying hens

AIMS: To investigate the prevalence of lead exposure in hens and eggs from backyard poultry in a sample of Auckland households, the relationship between concentrations of lead in the blood of the hens and in the shells and yolks of eggs from the same household, and to examine associations with measures of hen health, environment and husbandry factors.

METHODS: Thirty households participated in the study from August to November 2016, each providing one adult hen for sampling, an egg from the household if available, and completing a questionnaire on hen husbandry. Concentrations of lead in blood were determined using a portable lead analyser. Eggs were analysed for concentrations of lead in the yolk and shell using inductively coupled plasma mass spectrometry after biological digestion with a mixture of nitric and hydrochloric acid.

RESULTS: Twenty three of 30 hens (77%) showed evidence of lead exposure, with median concentrations of lead in blood of 0.77 (min?<0.16, max 8.02) μmol/L. All eggs showed evidence of lead exposure, with concentrations of lead in the yolk ranging from 0.003–1.07?mg/kg, and concentrations of lead in the eggshell ranging from <0.1–0.82?mg/kg. A positive correlation existed between concentrations of lead in the blood of a hen and concentrations of lead in egg yolk from the same hen (R²=0.97), and both the yolk (R²=0.58) and shell (R²=0.30) of an egg from her flock. No association was found between concentrations of lead in blood and hen health indices measured in this study. Concentrations of lead in blood were higher in hens from properties with homes built before 1941 than between 1941–1960 (p=0.03), and in hens from properties with weatherboard homes than brick homes (p=0.049).

CONCLUSIONS AND CLINICAL RELEVANCE: There was a high prevalence of lead exposure in this sample of Auckland backyard chickens, with the majority of hens being sub-clinically affected. Associations were found between concentrations of lead in the blood of the hens, and properties with homes built before 1941 and clad in weatherboard. Concentrations of lead in over half the egg yolks sampled were at levels sufficient to warrant human health concern. The assessment of concentrations of lead in backyard poultry and eggs intended for human consumption is recommended to protect human and bird health.     

Residues of gentamicin in eggs following medication of laying hens

1. Gentamicin was injected subcutaneously and intramuscularly into 5 groups of 10 laying hens and its concentration was determined in albumen, yolk and whole egg. 2. Groups 1 and 3 were intramuscularly injected with doses of 10 and 25 mg/kg while groups 2, 4 and 5 were subcutaneously injected with doses of 10, 25 and 50 mg/kg, respectively. 3. The final gentamicin concentration in albumen was measured on d 3 for groups 1 and 2; on d 4 for groups 3 and 4, and on d 5 for group 5. Concentrations in yolk and whole egg were measured on d 7, 10 and 12. 4. Gentamicin recovery was as follows: 2% in groups 1 and 2, 2.5% in groups 3 and 4, and 3% in group 5. 5. Most of the residue (approximately 90%) was recovered from the yolk.     

Distribution of 15N-chlorocholine chloride in eggs of laying hens

The distribution of Chlorocholine chloride (CCC) in the eggs of laying hens was studied using 15N-CCC. Twelve layers (37 weeks old) were divided into four groups and used in this study consisting of three feeding phases. In phase one (7 days), all the hens received a CCC-free diet [165 g CP/kg dry matter (DM); 11.58 MJ ME/kg DM]. In phase two (11 days), four levels of 15N-CCC: 0, 5, 50 and 250 ppm were added to the respective diets, while in phase three (7 days), CCC-free feed was again offered. Egg samples were taken and the 15N content of egg yolk and albumin were determined. At the end of phase two, there was a significant (p < 0.05) increase in 15N content in egg yolk from hens fed the 50 and 250 ppm CCC diets and in albumin from hens fed the 250 ppm CCC diet. The estimated 15N-CCC residue was 1.71, 6.64, 28.80 ppm in egg yolk and 1.58, 1.08 and 4.50 ppm in albumin from hens fed 5, 50 and 250 ppm CCC, respectively. The CCC residue, from quantitative analysis ranged from 0.21 to 0.93 and 0.93 to 2.43 ppm in yolk of hens fed 50 and 250 ppm CCC, respectively, whereas a range of 0.40-1.46 ppm, was found in the albumin of hens fed 250 ppm. The difference in measured CCC in yolk and albumin and that estimated from 15N-CCC could have been due to breakdown products of 15N-CCC. Seven days after withdrawal of 15N-CCC, the estimated 15N-CCC residue in egg yolk decreased to 0.43, 2.45 and 15.59 ppm, on 5, 50 and 250 ppm CCC dietary treatments, respectively, and to 2.46 ppm in albumin from hens fed 250 ppm CCC. The higher increase in 15N content could have been due to a higher incorporation of 15N-CCC into yolk than albumin during the process of rapid yolk deposition. This experiment showed that consumed CCC is distributed both into yolk and albumin in a dose dependent manner and that CCC is metabolized in laying hens. However, the level of CCC in the diet which could lead to accumulation of detectable CCC levels in eggs as observed in this study, is much higher than the established maximum residual limits in grains.     

Residues of dimetridazole in eggs after treatment of laying hens

Laying hens were dosed orally with dimetridazole (DMZ) (50 and 250 mg/kg) for 3 days or intramuscularly (50 mg/kg), also for 3 days, and the residues were determined by liquid chromatography in albumen and yolk. The sensitivity of the whole procedure was 2 ng/g. The drug was excreted preferentially into the yolk (about 57% of the total) and the elimination period lasted for 4–6 days after treatment.Abbreviations AUC area under the plasma concentration-time curve - depletion time the time needed for the DMZ concentration to fall below 0.01 g/g - elimination rate constant - Cl clearance - DMZ demetridazole     

Depletion of colistin in eggs following medication of laying hens

The depletion of colistin in eggs was determined separately for the albumen, the yolk and the whole egg after oral and intramuscular administrations of colistin sulphate. Residues were assayed by an agar plate diffusion method with Bordetella bronchoseptica ATCC 4617 as test organism. Colistin residues were not detected after drug administration by the oral route, but could be detected in the yolk until eight days after intramuscular injection. The total amount excreted represented 0.9% of the dose applied.     

Isolation of infectious bronchitis virus from intestine and reproductive organs of laying hens with dropped egg production

A total of 52 samples (22 cloacal swabs and 30 pooled ovaries and oviducts) from hens with a history of a 25 to 40% drop in egg production from 30 flocks were examined for the isolation of infectious bronchitis virus in embryonated chicken eggs. Five isolates were obtained: three from cloacal swabs and two from reproductive organs. These isolates were identified and characterized on the basis of agar-gel-precipitation, hemagglutination, and neutralization tests and other physicochemical characteristics.     

Serological detection of experimental Salmonella enteritidis infections in laying hens

The antibody response of laying hens to experimental Salmonella enteritidis infection was evaluated in microagglutination, tube agglutination, and rapid whole-blood plate agglutination assays. Hens of three different ages were infected by either oral inoculation or horizontal contact transmission. Blood was collected at weekly intervals, and the presence of specific antibodies was assessed by reaction with antigens prepared from strains of S. enteritidis and S. pullorum. The sensitivity of detection of infected hens did not vary significantly between the assays, as all three tests effectively identified most exposed hens as seropositive. Within each test, however, variation was observed in the detection sensitivity when different antigens were used. The microagglutination titers of serum samples were determined by serial dilution. Antibody titers peaked at 1 to 2 weeks postinoculation and declined steadily, although most birds were still identified as seropositive at 10 weeks postinoculation. The mean microtest titers obtained with S. enteritidis antigens were higher than with an S. pullorum antigen, indicating greater test sensitivity. However, use of the S. pullorum antigen resulted in fewer false positives when sera from uninfected control hens were tested. The titers of contact-exposed hens peaked later and at lower values than did those of inoculated hens, but these two groups of hens had similar antibody titers after the third week postinoculation.     

Residues of macrolide antibiotics in eggs following medication of laying hens

1. The elimination kinetics of four macrolide antibiotics (tylosin, erythromycin, spiramycin and josamycin) in eggs were determined separately for albumen and yolk after oral administration through either drinking water or diet or after intramuscular injection. 2. Residues were assayed by a plate diffusion technique in cylinders with Micrococcus luteus as the test-organism. 3. Drug excretion was usually over a longer time in the yolk. Spiramycin was the most highly excreted in the egg whereas seven to eight times less tylosin and erythromycin was transferred. The conditions for the use of macrolide antibiotics in laying hens are discussed.     

无公害蛋鸡与鸡蛋生产技术要点

无公害蛋鸡与鸡蛋，是指产地环境、生产过程和最终产品符合国家无公害食品标准和规范，经专门机构认定，按照国家《无公害农产品管理办法》的规定，许可使用无公害农产品标识的产品。无公害蛋鸡与鸡蛋符合国家食品卫生标准，具有无污染、安全、优质及营养的特点。随着加入WTO和北京、上海等大中城市陆续实行农     

Residues of aminoglycoside antibiotics in eggs after medication of laying hens

1. The transfer of aminoglycoside antibiotics into eggs was determined separately from albumen, yolk or whole egg after oral administration of dihydrostreptomycin (DHS), neomycin and spectinomycin and after an intramuscular injection of DHS. Residues were assayed by an agar plate diffusion method in cylinders with a specific test organism for each antibiotic. 2. Only DHS, administered by the intramuscular route, led to detectable residues in eggs. The total amount of DHS excreted via the eggs represented 1% of the dose administered. 3. Residues in the whole egg were detected for 8 d.