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Allostatic load refers to wear and tear on the body due to repeated activation of the stress response and, thus, may be an early subclinical indicator of future disease and mortality risk. To date, few studies of allostatic load have focused on young adults, racial/ethnic comparisons that include Mexican Americans, or the interplay between race/ethnicity, gender, and educational attainment. To fill these gaps, we used data on non-Hispanic Black, non-Hispanic White, and Mexican-origin respondents from Waves I (1994-1995) and IV (2007-2008) of the National Longitudinal Study of Adolescent to Adult Health (Add Health; N = 11,807). We calculated allostatic load scores based on respondents' values for 10 metabolic, cardiovascular, and inflammatory biomarkers measured at Wave IV, when respondents were 24-34 years old. We then used negative binomial regression models to assess the combined effects of race/ethnicity, gender, and educational attainment on allostatic load, while controlling for key covariates. We found that Black women had significantly higher allostatic load scores than White women and Black men, net of educational attainment and other covariates. Yet, education modified the relationship between race/ethnicity, gender, and allostatic load. Obtaining a college education was protective for White males and females but no more or less protective for other women and deleterious for Black males. In other words, by the time they reach young adulthood, the cumulative physiological burden of stress on Black women and college-educated Black men is already greater than it is among their similarly or less educated White counterparts. These findings provide important information about the intermediate physiological dysregulation that underlies social inequalities in stress-related health outcomes, especially those that occur at the intersections of race/ethnicity, gender, and educational attainment. They also suggest that research on its antecedents should focus on earlier life periods., Competing Interests: None., (© 2021 The Author(s).)

The Pfam database is a widely used resource for classifying protein sequences into families and domains. Since Pfam was last described in this journal, over 350 new families have been added in Pfam 33.1 and numerous improvements have been made to existing entries. To facilitate research on COVID-19, we have revised the Pfam entries that cover the SARS-CoV-2 proteome, and built new entries for regions that were not covered by Pfam. We have reintroduced Pfam-B which provides an automatically generated supplement to Pfam and contains 136 730 novel clusters of sequences that are not yet matched by a Pfam family. The new Pfam-B is based on a clustering by the MMseqs2 software. We have compared all of the regions in the RepeatsDB to those in Pfam and have started to use the results to build and refine Pfam repeat families. Pfam is freely available for browsing and download at http://pfam.xfam.org/., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Purpose: Although peer support has received research attention within different health related contexts, there is limited research considering individuals who have experienced an amputation. In particular, the peer mentoring role is under-explored. Therefore, this research aimed to explore the experiences of participants delivering peer support interventions to individuals with lower limb loss. Methods: Eight people who acted as peer mentors for people with limb loss took part in semi-structured interviews. Data were analyzed using Interpretative Phenomenological Analysis (IPA). Results: Two overarching themes were identified from the data: "Developing a Helpful Self", in which the personal value and meaning of being a peer mentor is presented, and "Connecting with Vulnerability", which addresses the emotional challenges of peer mentoring and the impact of these on wellbeing. Conclusions: Findings suggest positive experiences of providing support, including increased hope, resilience and a sense of belonging and connection to others. However, peer mentors experienced challenges related to uncertainty and doubt about their mentoring abilities and with developing and maintaining resilience in the role. Recommendations include the development of training packages and increased clarity for volunteers of the peer mentor role.Implications for rehabilitationIt is important to define clearly the role of a peer mentor and the responsibilities this involves.Peer mentors should be provided with support and training to increase their confidence and ability to act appropriately when encountering distress.It is important to provide transparent guidelines and procedures to support peer mentors to minimize concerns over risk and safety.Feedback regarding how effective the support offered is and how it could be improved should be provided.

Background: As of August 11, 2020, Coronavirus disease 2019 (COVID-19) has infected 19,936,210 persons and led to 732,499 deaths worldwide. The impact has been immense, and with no vaccine currently available, the best way to protect our communities is health education. We developed a brief COVID-19 knowledge test for health educators that can be used to assess deficits in clients' understanding of the disease. Methods: COVID-19 Knowledge Test items were developed by the research team and administered to participants. An alternate-choice item format was selected for the knowledge test, and data analysis was based on an American sample of 273 respondents. A detailed analysis of the data was conducted with classical test theory and Rasch analysis. Findings: The final instrument was found to be a unidimensional measure of COVID-19 knowledge. Results provided evidence for absolute model fit and model fit for individual items. All items included on the scale were monotonically increasing and split-half reliability was considered acceptable. Total test information revealed that the test is suitable for individuals with low to average knowledge of COVID-19. Interpretation: Rasch analysis provides support for the COVID-19 Knowledge Test to be used as an assessment tool for health educators. The final version of the test consists of 34 high-quality test items that can be administered in <10 min. Normative data and suggested cutoff scores are also provided., (Copyright © 2020 Richardson and Bélanger.)

MGnify (http://www.ebi.ac.uk/metagenomics) provides a free to use platform for the assembly, analysis and archiving of microbiome data derived from sequencing microbial populations that are present in particular environments. Over the past 2 years, MGnify (formerly EBI Metagenomics) has more than doubled the number of publicly available analysed datasets held within the resource. Recently, an updated approach to data analysis has been unveiled (version 5.0), replacing the previous single pipeline with multiple analysis pipelines that are tailored according to the input data, and that are formally described using the Common Workflow Language, enabling greater provenance, reusability, and reproducibility. MGnify's new analysis pipelines offer additional approaches for taxonomic assertions based on ribosomal internal transcribed spacer regions (ITS1/2) and expanded protein functional annotations. Biochemical pathways and systems predictions have also been added for assembled contigs. MGnify's growing focus on the assembly of metagenomic data has also seen the number of datasets it has assembled and analysed increase six-fold. The non-redundant protein database constructed from the proteins encoded by these assemblies now exceeds 1 billion sequences. Meanwhile, a newly developed contig viewer provides fine-grained visualisation of the assembled contigs and their enriched annotations., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Automatic annotation of protein function is routinely applied to newly sequenced genomes. While this provides a fine-grained view of an organism's functional protein repertoire, proteins, more commonly function in a coordinated manner, such as in pathways or multimeric complexes. Genome Properties (GPs) define such functional entities as a series of steps, originally described by either TIGRFAMs or Pfam entries. To increase the scope of coverage, we have migrated GPs to function as a companion resource utilizing InterPro entries. Having introduced GPs-specific versioned releases, we provide software and data via a GitHub repository, and have developed a new web interface to GPs (available at https://www.ebi.ac.uk/interpro/genomeproperties). In addition to exploring each of the 1286 GPs, the website contains GPs pre-calculated for a representative set of proteomes; these results can be used to profile GPs phylogenetically via an interactive viewer. Users can upload novel data to the viewer for comparison with the pre-calculated results. Over the last year, we have added ∼700 new GPs, increasing the coverage of eukaryotic systems, as well as increasing general coverage through automatic generation of GPs from related resources. All data are freely available via the website and the GitHub repository.

The last few years have witnessed significant changes in Pfam (https://pfam.xfam.org). The number of families has grown substantially to a total of 17,929 in release 32.0. New additions have been coupled with efforts to improve existing families, including refinement of domain boundaries, their classification into Pfam clans, as well as their functional annotation. We recently began to collaborate with the RepeatsDB resource to improve the definition of tandem repeat families within Pfam. We carried out a significant comparison to the structural classification database, namely the Evolutionary Classification of Protein Domains (ECOD) that led to the creation of 825 new families based on their set of uncharacterized families (EUFs). Furthermore, we also connected Pfam entries to the Sequence Ontology (SO) through mapping of the Pfam type definitions to SO terms. Since Pfam has many community contributors, we recently enabled the linking between authorship of all Pfam entries with the corresponding authors' ORCID identifiers. This effectively permits authors to claim credit for their Pfam curation and link them to their ORCID record.

The InterPro database (http://www.ebi.ac.uk/interpro/) classifies protein sequences into families and predicts the presence of functionally important domains and sites. Here, we report recent developments with InterPro (version 70.0) and its associated software, including an 18% growth in the size of the database in terms on new InterPro entries, updates to content, the inclusion of an additional entry type, refined modelling of discontinuous domains, and the development of a new programmatic interface and website. These developments extend and enrich the information provided by InterPro, and provide greater flexibility in terms of data access. We also show that InterPro's sequence coverage has kept pace with the growth of UniProtKB, and discuss how our evaluation of residue coverage may help guide future curation activities.

Historically, intersectionality has been an underutilized framework in sociological research on racial/ethnic and gender inequalities in health. To demonstrate its utility and importance, we conduct an intersectional analysis of the social stratification of health using the exemplar of hypertension-a health condition in which racial/ethnic and gender differences have been well-documented. Previous research has tended to examine these differences separately and ignore how the interaction of social status dimensions may influence health over time. Using seven waves of data from the Health and Retirement Study and multilevel logistic regression models, we found a multiplicative effect of race/ethnicity and gender on hypertension risk trajectories, consistent with both an intersectionality perspective and persistent inequality hypothesis. Group differences in past and contemporaneous socioeconomic and behavioral factors did not explain this effect.

This study examines how the intersecting consequences of race-ethnicity, gender, socioeconomics status (SES), and age influence health inequality. We draw on multiple-hierarchy stratification and life course perspectives to address two main research questions. First, does racial-ethnic stratification of health vary by gender and/or SES? More specifically, are the joint health consequences of racial-ethnic, gender, and socioeconomic stratification additive or multiplicative? Second, does this combined inequality in health decrease, remain stable, or increase between middle and late life? We use panel data from the Health and Retirement Study (N = 12,976) to investigate between- and within-group differences in in self-rated health among whites, blacks, and Mexican Americans. Findings indicate that the effects of racial-ethnic, gender, and SES stratification are interactive, resulting in the greatest racial-ethnic inequalities in health among women and those with higher levels of SES. Furthermore, racial-ethnic/gender/SES inequalities in health tend to decline with age. These results are broadly consistent with intersectionality and aging-as-leveler hypotheses., (© American Sociological Association 2016.)

A short-term program of performing serial active screening cultures (ASC) in the intensive care unit was instituted to establish a method for the detection of antibiotic-resistant Gram-negative bacteria (GNB) and the local rates of colonisation. Of all submitted ASC, 25.9% (30/116 collected swabs) isolated an antibiotic-resistant GNB. ChromID ESBL agar (bioMérieux, France) identified the majority of these organisms, with the additional antibiotic-impregnated media [MacConkey agar (MCA) with ciprofloxacin, MCA with gentamicin and MCA with ceftazidime] adding limited benefit. Compared to swabs performed on admission, 37.8% (14/37) of patients cultured a new antibiotic-resistant isolate on discharge. Serial screening in intensive care has the ability to identify patients with unrecognised colonisation with antibiotic-resistant GNB; however, the increase in the laboratory workload and logistical challenges in the collection of the surveillance swabs may limit this program's expansion.

Background: With the emergence of obesity as a global health issue an increasing number of major demographic surveys are collecting measured anthropometric data. Yet little is known about the characteristics and reliability of these data., Objectives: We evaluate the accuracy and reliability of anthropometric data collected in the home during Wave IV of the National Longitudinal Study of Adolescent to Adult Health (Add Health), compare our estimates to national standard, clinic-based estimates from the National Health and Nutrition Examination Survey (NHANES) and, using both sources, provide a detailed anthropometric description of young adults in the United States., Methods: The reliability of Add Health in-home anthropometric measures was estimated from repeat examinations of a random subsample of study participants. A digit preference analysis evaluated the quality of anthropometric data recorded by field interviewers. The adjusted odds of obesity and central obesity in Add Health vs. NHANES were estimated with logistic regression., Results: Short-term reliabilities of in-home measures of height, weight, waist and arm circumference-as well as derived body mass index (BMI, kg/m 2 )-were excellent. Prevalence of obesity (37% vs. 29%) and central obesity (47% vs. 38%) was higher in Add Health than in NHANES while socio-demographic patterns of obesity and central obesity were comparable in the two studies., Conclusions: Properly trained non-medical field interviewers can collect reliable anthropometric data in a nationwide, home visit study. This national cohort of young adults in the United States faces a high risk of early-onset chronic disease and premature mortality.

In North America and Europe, the binary toxin positive Clostridium difficile strains of the ribotypes 027 and 078 have been associated with death, toxic megacolon and other adverse outcomes. Following an increase in C. difficile infections (CDIs) in Queensland, a prevalence study involving 175 hospitals was undertaken in early 2012, identifying 168 cases of CDI over a 2 month period. Patient demographics and clinical characteristics were recorded, and C. difficile isolates were ribotyped and tested for the presence of binary toxin genes. Most patients (106/168, 63.1%) were aged over 60 years. Overall, 98 (58.3%) developed symptoms after hospitalisation; 89 cases (53.0%) developed symptoms more than 48 hours after admission. Furthermore, 27 of the 62 (67.7%) patients who developed symptoms in the community ad been hospitalised within the last 3 months. Thirteen of the 168 (7.7%) cases identified had severe disease, resulting in admission to the Intensive Care Unit or death within 30 days of the onset of symptoms. The 3 most common ribotypes isolated were UK 002 (22.9%), UK 014 (13.3%) and the binary toxin-positive ribotype UK 244 (8.4%). The only other binary toxin positive ribotype isolated was UK 078 (n = 1). Of concern was the detection of the binary toxin positive ribotype UK 244, which has recently been described in other parts of Australia and New Zealand. No isolates were of the international epidemic clone of ribotype UK 027, although ribotype UK 244 is genetically related to this clone. Further studies are required to track the epidemiology of ribotype UK 244 in Australia and New Zealand., (This work is copyright. You may download, display, print and reproduce the whole or part of this work in unaltered form for your own personal use or, if you are part of an organisation, for internal use within your organisation, but only if you or your organisation do not use the reproduction for any commercial purpose and retain this copyright notice and all disclaimer notices as part of that reproduction. Apart from rights to use as permitted by the Copyright Act 1968 or allowed by this copyright notice, all other rights are reserved and you are not allowed to reproduce the whole or any part of this work in any way (electronic or otherwise) without first being given the specific written permission from the Commonwealth to do so. Requests and inquiries concerning reproduction and rights are to be sent to the Online, Services and External Relations Branch, Department of Health, GPO Box 9848, Canberra ACT 2601, or by email to copyright@health.gov.au.)

Background: This analysis explored the effect of timing, sequencing, and change in preconception health across adolescence and young adulthood on racial/ethnic disparities in birth weight in a diverse national cohort of young adult women., Methods: Data came from Waves I (1994-1995), III (2001-2002), and IV (2007-2008) of the National Longitudinal Study of Adolescent Health. Eligibility was restricted to all singleton live births to female non-Hispanic White, non-Hispanic Black, Mexican-origin Latina, or Asian/Pacific Islander participants (n = 3,014) occurring between the Wave III (ages 18-26 years) and IV (ages 24-32 years) interviews. Birth weight was categorized into low (<2,500 g), normal (2,500-4,000 g), and macrosomic (>4,000 g). Preconception health indicators were cigarette smoking, heavy alcohol consumption, overweight or obesity, and inadequate physical activity, measured in adolescence (Wave I, ages 11-19 years) and early adulthood (Wave III) and combined into four-category variables to capture the timing and sequencing of exposure., Findings: Measures of preconception health did not explain the Black-White disparity in low birth weight, which increased after adjustment for confounders (odds ratio [OR], 2.17; 95% confidence interval [CI], 1.33-3.53) and effect modification by overweight/obesity (OR, 3.58; 95% CI, 1.65-7.78). A positive association between adult-onset overweight/obesity and macrosomia was modified by race (OR, 3.83; 95% CI, 1.02-14.36 for Black women)., Conclusions: This longitudinal analysis provides new evidence on preconception health and racial/ethnic disparities in birth weight. Specifically, it indicates that interventions focused on prevention of overweight/obesity and maintenance of healthy weight during the transition to adulthood, especially among Black females, may be warranted., (Copyright © 2014 Jacobs Institute of Women's Health. Published by Elsevier Inc. All rights reserved.)

In the tropical city of Darwin, Northern Territory, Australia, dry season soil sampling cultured Burkholderia pseudomallei from 7 (70%) of 10 sports fields. However, during the 23 years of the Darwin Prospective Melioidosis Study, only 5 (0.6%) of 785 melioidosis cases have been attributed to infection from sports fields. In one soccer player with cutaneous melioidosis, B. pseudomallei cultured from the player was identical by multilocus sequence typing and multilocus variable-number tandem repeat analysis with an isolate recovered from soil at the location on the sports field where he was injured. Melioidosis is uncommon in otherwise healthy sports persons in melioidosis-endemic regions but still needs consideration in persons with abrasion injuries that involve contact with soil.

Objective. This study examined to what degree patient-centeredness-measured as an underlying ability of obstetrical nurses-influenced Medicaid patients' satisfaction with care in hospital obstetrical units. Design. Multigroup structural equation modeling design, using three cross-sectional random samples (n = 300 each) from the 2003 Press Ganey National Inpatient Database. Setting. Self-administered mail surveys. Participants. 900 Medicaid recipients recently discharged from inpatient hospital obstetrical units across the United States. Methods. Multigroup structural equation modeling was used to test the goodness of fit between a hypothesized model based on the Primary Provider Theory and patients' ratings of nurses. Results. The model fitted the data well, was stable across three random samples, and was sustained when compared to a competing model. The patient-centeredness of nurses significantly influenced overall patient satisfaction and explained 66% of its variability. When nurses' patient-centeredness increased by one standard deviation, patients' satisfaction increased by 0.80 standard deviation. Conclusion. This study offers a novel approach to the measurement of the patient-centeredness of nurses and a paradigm for increasing it and its influence on Medicaid patients' satisfaction in hospital obstetrical units.

Purpose: This study was designed to assess the relationship between birth weight and prospectively measured trajectories of preconception health across adolescence and young adulthood in a diverse national cohort of young adult women., Methods: Data came from Waves I (1994-1995), III (2001-2002), and IV (2007-2008) of the National Longitudinal Study of Adolescent Health. Eligibility was restricted to all the singleton live births (n = 3,436) to female participants occurring between the Wave III (ages 18-26 years) and Wave IV (ages 24-32 years) interviews. Preconception cigarette smoking, overweight/obesity, adequate physical activity, heavy alcohol consumption, and fair/poor self-rated health were measured in adolescence (Wave I) and early adulthood (Wave III) and combined into four-category variables to capture the timing and sequencing of exposure. The outcome measure, birth weight, was classified as low (<2,500 g), normal (2,500-4,000 g), and macrosomic (>4,000 g)., Results: Multinomial logistic regression results indicated that adult-onset overweight significantly increased the odds of having a macrosomic birth (odds ratio = 1.56; 95% confidence interval = 1.02-2.38)., Conclusions: This study provides new evidence about the influence of maternal body mass index trajectories on offspring birth weight. Adult-onset overweight/obesity during the transition to adulthood was common in the sample and increased the odds of subsequently delivering a macrosomic infant by 56%. This finding suggests that healthy weight promotion before this transition would confer intergenerational benefits, and supports recommendations for preconception care to address overweight/obesity., (Copyright © 2012 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.)

Campylobacter is an important human pathogen, and consumption of undercooked poultry has been linked to significant human illnesses. To reduce human illness, intervention strategies targeting Campylobacter reduction in poultry are in development. For more than a decade, there has been an ongoing national and international controversy about whether Campylobacter can pass from one generation of poultry to the next via the fertile egg. We recognize that there are numerous sources of Campylobacter entry into flocks of commercial poultry (including egg transmission), yet the environment is often cited as the only source. There has been an abundance of published research globally that refutes this contention, and this article lists and discusses many of them, along with other studies that support environment as the sole or primary source. One must remember that egg passage can mean more than vertical, transovarian transmission. Fecal bacteria, including Campylobacter, can contaminate the shell, shell membranes, and albumen of freshly laid fertile eggs. This contamination is drawn through the shell by temperature differential, aided by the presence of moisture (the "sweating" of the egg); then, when the chick emerges from the egg, it can ingest bacteria such as Campylobacter, become colonized, and spread this contamination to flock mates in the grow house. Improvements in cultural laboratory methods continue to advance our knowledge of the ecology of Campylobacter, and in the not-so-distant future, egg passage will not be a subject continuously debated but will be embraced, thus allowing the development and implementation of more effective intervention strategies.

Melioidosis is an emerging infectious disease caused by the soil bacterium Burkholderia pseudomallei. In diagnostic and forensic settings, molecular detection assays need not only high sensitivity with low limits of detection but also high specificity. In a direct comparison of published and newly developed TaqMan PCR assays, we found the TTS1-orf2 assay to be superior in detecting B. pseudomallei directly from clinical specimens. The YLF/BTFC multiplex assay (targeting the Yersinia-like fimbrial/Burkholderia thailandensis-like flagellum and chemotaxis region) also showed high diagnostic sensitivity and provides additional information on possible geographic origin.

Optimising DNA extraction from clinical samples for Burkholderia pseudomallei Type III secretion system real-time PCR in suspected melioidosis patients confirmed that urine and sputum are useful diagnostic samples. Direct testing on blood remains problematic; testing DNA extracted from plasma was superior to DNA from whole blood or buffy coat., (Copyright © 2011 Elsevier B.V. All rights reserved.)

This study was conducted to evaluate the influence of a vancomycin pretreatment on the ability of marker (nalidixic-acid resistant) Salmonella Enteritidis (SE(M)), field Salmonella Enteritidis (SE(E)), and marker Salmonella Typhimurium (ST(M)) strains to colonize within the intestinal and reproductive tracts and translocate to other organs of leghorn laying hens. In each of three trials, caged laying hens (76, 26, and 33 wk ofage) were divided into six groups designated to receive SE(M), SE(F), or ST(M), and half were pretreated with vancomycin (n = 11-12 hens). Vancomycin-treated hens received 10 mg vancomycin in saline/kilogram body weight orally for 5 days to inhibit Gram-positive bacteria within the intestines. On Day 6, all hens were concurrently challenged by oral, intravaginal, and intracolonal routes with Salmonella and placed into separate floor chambers by Salmonella strain. Two weeks postinoculation, all hens were euthanatized and the ceca, spleen, liver/gall bladder (LGB), upper (URT), and lower (LRT) reproductive tracts, and ovarian follicles were aseptically collected, and analyzed for Salmonella. Results did not differ for the three hen's ages and were therefore combined. The vancomycin pretreatment also had no significant effect on the colonization ability of SE(M), SE(F) or ST(M), and therefore results were combined within Salmonella strain. The marker strain of Salmonella Enteritidis was recovered from 21% of ceca, 4% of LGB, 9% of LRT, and 17% of the fecal samples. The field strain of Salmonella Enteritidis was recovered from 88% of ceca, 96% of spleen, 92% of LGB, 30% of LRT, 4% of URT, 13% of follicle, and 42% of the fecal samples. The marker strain of Salmonella Typhimurium was recovered from 100% of ceca, 74% of spleen, 91% of LGB, 30% of LRT, 9% of URT, 9% of follicle, and 100% of the fecal samples. Among ceca, spleen, LGB, and fecal samples, SE(F) and ST(M) colonization was significantly greater than SE(M) colonization. Overall prevalence of Salmonella in the reproductive tracts of challenged hens was relatively low, ranging from 4% to 30%.

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.

Data relating to acute post-streptococcal glomerulonephritis (APSGN) from the notifiable diseases surveillance system in the Northern Territory of Australia was extracted and analyzed. Isolates of Streptococcus pyogenes from confirmed cases were emm sequence typed. From 1991 to July 2008, there were 415 confirmed cases and 23 probable cases of APSGN notified. Four hundred fifteen (94.7%) of these were Indigenous Australians and 428 (97.7%) were people living in remote or very remote locations. The median age of cases was 7 years (range 0-54). The incidence of confirmed cases was 12.5/100,000 person-years, with an incidence in Indigenous Australian children younger than 15 years of age of 94.3 cases/100,000 person-years. The overall rate ratio of confirmed cases in Indigenous Australians to non-Indigenous Australians was 53.6 (95% confidence interval 32.6-94.8). Outbreaks of disease across multiple communities occurred in 1995 (N = 68), 2000 (N = 55), and 2005 (N = 87 [confirmed cases]). Various emm types of S. pyogenes were isolated from cases of APSGN including some types not previously recognized to be nephritogenic. The widespread outbreak in 2005 was caused by emm55.0 S. pyogenes. Acute post-streptococcal glomerulonephritis continues to occur in remote Indigenous communities in Australia at rates comparable to or higher than those estimated in developing countries. Improvements in preventative and outbreak control strategies are needed.

Campylobacter spp. are present in organs and tissues of broiler chickens but the dissemination route is unclear. The aim of the current study was to determine Campylobacter prevalence within circulating blood of commercial broilers. Broilers were acquired from 19 flocks originating from three commercial poultry processing companies. Using aseptic blood collection techniques, 5 ml of circulating blood was collected from each bird and the sample analyzed for Campylobacter. The Campylobacter colonization status of each bird was determined by aseptically sampling and analyzing the ceca. Campylobacter was recovered from 58% (11/19) of flocks sampled. From the 248 total birds sampled, 12% and 46% of the birds had Campylobacter in the blood and ceca, respectively. This study documents Campylobacter prevalence in the circulating blood of commercially raised broilers. Campylobacter presence in the circulatory system may indicate the path used by the organism for rapid dissemination to organs and tissues. From a processing viewpoint, Campylobacter presence in circulating blood of market-age broilers may increase the likelihood of cross-contamination between birds during slaughter.

The major limitation of current typing methods for Streptococcus pyogenes, such as emm sequence typing and T typing, is that these are based on regions subject to considerable selective pressure. Multilocus sequence typing (MLST) is a better indicator of the genetic backbone of a strain but is not widely used due to high costs. The objective of this study was to develop a robust and cost-effective alternative to S. pyogenes MLST. A 10-member single nucleotide polymorphism (SNP) set that provides a Simpson's Index of Diversity (D) of 0.99 with respect to the S. pyogenes MLST database was derived. A typing format involving high-resolution melting (HRM) analysis of small fragments nucleated by each of the resolution-optimized SNPs was developed. The fragments were 59-119 bp in size and, based on differences in G+C content, were predicted to generate three to six resolvable HRM curves. The combination of curves across each of the 10 fragments can be used to generate a melt type (MelT) for each sequence type (ST). The 525 STs currently in the S. pyogenes MLST database are predicted to resolve into 298 distinct MelTs and the method is calculated to provide a D of 0.996 against the MLST database. The MelTs are concordant with the S. pyogenes population structure. To validate the method we examined clinical isolates of S. pyogenes of 70 STs. Curves were generated as predicted by G+C content discriminating the 70 STs into 65 distinct MelTs., (© 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.)

Purpose: We evaluated the contributions of birth weight and current body mass index (BMI) to racial/ethnic disparities in systolic blood pressure (SBP) in the United States., Methods: Participants were 10,046 young adults (ages 24-32) in the National Longitudinal Study of Adolescent Health. SBP, BMI, and other contemporaneous factors were assessed at Wave IV (2007-2008); birth weight and other early life factors were reported at Wave I (1994-1995). Data were analyzed using sex- and race-stratified multivariable regression models., Results: Racial/ethnic disparities in SBP were limited to black and white females. The black-white female disparity in SBP was 3.36 mmHg and was partially explained by current BMI, but not birth weight. Associations between birth weight and SBP were limited to males, in whom we found a decrease of 1.05 mmHg in SBP per 1-kg increase in birth weight (95% confidence interval, -1.90, -0.20). This inverse relationship strengthened after adjusting for BMI and other factors, and was strongest among black and white males. A significant association between BMI and SBP was found in all racial/ethnic and sex subgroups., Conclusions: In this U.S. national cohort, birth weight is negatively associated with SBP among black and white young adult males., (Copyright © 2011 Elsevier Inc. All rights reserved.)

These studies evaluated the bacterial level of unwashed and washed shell eggs from caged and cage-free laying hens. Hy-Line W-36 White and Hy-Line Brown laying hens were housed on all wire slats or all shavings floor systems. On the sampling days for experiments 1, 2, and 3, 20 eggs were collected from each pen for bacterial analyses. Ten of the eggs collected from each pen were washed for 1 min with a commercial egg-washing solution, whereas the remaining 10 eggs were unwashed before sampling the eggshell and shell membranes for aerobic bacteria and coliforms (experiment 1 only). In experiment 1, the aerobic plate counts (APC) of unwashed eggs produced in the shavings, slats, and caged-housing systems were 4.0, 3.6, and 3.1 log(10) cfu/mL of rinsate, respectively. Washing eggs significantly (P < 0.05) reduced APC by 1.6 log(10) cfu/mL and reduced the prevalence of coliforms by 12%. In experiment 2, unwashed eggs produced by hens in triple-deck cages from 57 to 62 wk (previously housed on shavings, slats, and cages) did not differ, with APC ranging from 0.6 to 0.8 log(10) cfu/mL. Washing eggs continued to significantly reduce APC to below 0.2 log(10) cfu/mL. In experiment 3, the APC for unwashed eggs were within 0.4 log below the APC attained for unwashed eggs in experiment 1, although hen density was 28% of that used in experiment 1. Washing eggs further lowered the APC to 0.4 to 0.7 log(10) cfu/mL, a 2.7-log reduction. These results indicate that shell bacterial levels are similar after washing for eggs from hens housed in these caged and cage-free environments. However, housing hens in cages with manure removal belts resulted in lower APC for both unwashed and washed eggs (compared with eggs from hens housed in a room with shavings, slats, and cages).

Many reviews have discussed Salmonella in poultry and suggested best practices to minimize this organism on raw poultry meat. Despite years of research and conscientious control efforts by industry and regulatory agencies, human salmonellosis rates have declined only modestly and Salmonella is still found on raw poultry. Expert committees have repeatedly emphasized the importance of controlling risk, but information about Salmonella in poultry is often limited to prevalence, with inadequate information about testing methods or strains of Salmonella that are detected by these methods and no information about any impact on the degree of risk. This review examines some assumptions behind the discussion of Salmonella in poultry: the relationships between sampling and cultural methodology, prevalence and numbers of cells, and the implications of serotype and subtype issues. Minimizing Salmonella contamination of poultry is not likely to reduce human salmonellosis acquired from exposure to contaminated chicken until these issues are confronted more systematically.

We have developed a single nucleotide polymorphism (SNP) nucleated high-resolution melting (HRM) technique to genotype Enterococcus faecium. Eight SNPs were derived from the E. faecium multilocus sequence typing (MLST) database and amplified fragments containing these SNPs were interrogated by HRM. We tested the HRM genotyping scheme on 85 E. faecium bloodstream isolates and compared the results with MLST, pulsed-field gel electrophoresis (PFGE) and an allele specific real-time PCR (AS kinetic PCR) SNP typing method. In silico analysis based on predicted HRM curves according to the G+C content of each fragment for all 567 sequence types (STs) in the MLST database together with empiric data from the 85 isolates demonstrated that HRM analysis resolves E. faecium into 231 "melting types" (MelTs) and provides a Simpson's Index of Diversity (D) of 0.991 with respect to MLST. This is a significant improvement on the AS kinetic PCR SNP typing scheme that resolves 61 SNP types with D of 0.95. The MelTs were concordant with the known ST of the isolates. For the 85 isolates, there were 13 PFGE patterns, 17 STs, 14 MelTs and eight SNP types. There was excellent concordance between PFGE, MLST and MelTs with Adjusted Rand Indices of PFGE to MelT 0.936 and ST to MelT 0.973. In conclusion, this HRM based method appears rapid and reproducible. The results are concordant with MLST and the MLST based population structure.

Whether a required Salmonella test series is passed or failed depends not only on the presence of the bacteria but also on the methods for taking samples, the methods for culturing samples, and the statistics associated with the sampling plan. The pass-fail probabilities of the 2-class attribute sampling plans used for testing chilled chicken carcasses in the United States and Europe were compared by calculation and simulation. Testing in the United States uses whole-carcass rinses (WCR), with a maximum number of 12 positives out of 51 carcasses in a test set. Those numbers were chosen so that a plant operating with a Salmonella prevalence of 20%, the national baseline result for broiler chicken carcasses, has an approximately 80% probability of passing a test set. The European Union requires taking neck skin samples of approximately 8.3 g each from 150 carcasses, with the neck skins cultured in pools of 3 and with 7 positives as the maximum passing score for a test set of 50 composite samples. For each of these sampling plans, binomial probabilities were calculated and 100,000 complete sampling sets were simulated using a random number generator in a spreadsheet. Calculations indicated that a 20% positive rate in WCR samples was approximately equivalent to an 11.42% positive rate in composite neck skin samples or a 3.96% positive rate in individual neck skin samples within a pool of 3. With 20% as the prevalence rate, 79.3% of the simulated WCR sets passed with 12 or fewer positive carcasses per set, very near the expected 80% rate. Under simulated European conditions, a Salmonella prevalence of 3.96% in individual neck skin samples yielded a passing rate of 79.1%. The 2 sampling plans thus have roughly equivalent outcomes if WCR samples have a Salmonella-positive rate of 20% and individual neck skin samples have a positive rate of 3.96%. Sampling and culturing methods must also be considered in comparing the different standards for Salmonella.

There is a high burden of disease due to group A streptococcus (GAS) in remote Northern Territory (NT) Indigenous communities. A proposed 26-valent GAS M-type vaccine covers 80-90% of pharyngeal and invasive isolates in the US. We examined the diversity and distribution of emm types in two remote Indigenous communities in the NT Top End over a 17-year period and compared them to the proposed vaccine types. Eighty emm types were identified between 1991 and 2007. Diversity in both communities was high (overall Simpson's index 0.976), but varied between communities. Prior to 2004, 71 emm types were identified and an additional 9 emm types were identified during a period of active surveillance in 2004-2005. The proposed 26-valent vaccine would be expected to cover only 20% of emm types recovered in this study. Of the 80 emm types, 16 (20%) were new sequence types identified since the last assignment of M types in 2002. The diversity of streptococcal isolates was higher than that reported from most industrialized countries, and similar to that described in several developing countries. A vaccine based on such a variable antigen is unlikely to provide effective protection in the highest risk populations., ((c) 2010 Elsevier Ltd. All rights reserved.)

Sampling protocols for detecting Salmonella on poultry differ among various countries. In the United States, the U.S. Department of Agriculture Food Safety and Inspection Service dictates that whole broiler carcasses should be rinsed with 400 ml of 1% buffered peptone water, whereas in the European Union 25-g samples composed of neck skin from three carcasses are evaluated. The purpose of this study was to evaluate a whole carcass rinse (WCR) and a neck skin excision (NS) procedure for Salmonella and Escherichia coli isolation from the same broiler carcass. Carcasses were obtained from three broiler processing plants. The skin around the neck area was aseptically removed and bagged separately from the carcass, and microbiological analysis was performed. The corresponding carcass was bagged and a WCR sample was evaluated. No significant difference (alpha

Campylobacter spp. are frequently present in the intestinal tract and internal tissues of broiler breeder and broiler chickens. Campylobacter spp. ecology in commercial Leghorn laying hens has not been extensively studied. The objectives of the current study were to determine 1) Campylobacter spp. presence in the reproductive tract, lymphoid organs, liver-gallbladder, and ceca of commercial Leghorn laying hens; 2) species of Campylobacter present; and 3) antimicrobial resistance pattern of Campylobacter isolates. In study 1, three flocks ranging from 94 to 105 wk of age were sampled from a commercial laying complex. In study 2, two flocks, 82 and 84 wk of age, were sampled from a separate complex. Hens were killed, defeathered, aseptically necropsied, and the spleen, liver-gallbladder, ovarian follicles, and upper (infundibulum, magnum, and isthmus) and lower (shell gland and vagina) reproductive tracts were aseptically removed before the ceca. Samples were packed on ice and transported to the laboratory for evaluation. For speciation, a standard BAX real-time PCR method was used while susceptibility testing was performed using US National Antimicrobial Resistance Monitoring System (NARMS) standards and recommended quality control organisms. Isolates were examined for susceptibility using a semi-automated testing system (Sensititer) to the following 9 antimicrobials: azithromycin, clindamycin, ciprofloxacin, erythromycin, florfenicol, gentamicin, nalidixic acid, telithromycin, and tetracycline. In study 1, the isolation rate was 13, 67, 53, 3, 13, and 57% from the ovarian follicles, lower reproductive tract, upper reproductive tract, spleen, liver-gallbladder, and ceca, respectively. In study 2, the isolation rate was 17, 43, 33, 20, 17, and 73% from the ovarian follicles, lower reproductive tract, upper reproductive tract, spleen, liver-gallbladder, and ceca, respectively. Overall, 50% of isolates were Campylobacter jejuni, 49% Campylobacter coli, and 1% Campylobacter lari. In study 1, all of the isolates were pan-susceptible. In study 2, thirty-seven percent of the isolates were resistant to tetracycline. Commercial table egg laying hens housed in colony cages on wire floors had diverse Campylobacter spp. recovered from different tissues and these isolates were not resistant to a broad range of antimicrobials.

Campylobacter inoculation studies are limited without a suitable marker strain. The lurpose of this study was to screen Campylobacter strains (n=2073) obtained from poultry carcass rinses through the Centers for Disease Control and Prevention's National Antimicrobial Resistant Monitoring System for resistance to gentamicin and evaluate one strain's efficacy as a marker. A C. coli strain was found resistant to gentamicin at >32 microg/ml. Gentamicin was incorporated into media (Campy-Cefex agar, Brucella agar, and blood agar) from 0 to 1000 microg/ml, and the upper level of gentamicin resistance was determined. C. coli strain's upper level of growth on Campy-Cefex plates, blood agar plates, and Brucella agar plates was 400, 300, and 200 pg/ml, respectively. Ceca and postpick carcass rinses were obtained and streaked onto Campy-Cefex agar at the above gentamicin levels to evaluate background microflora exclusion. Campy-Cefex agar containing gentamicin at 100 ag/ml prevented from the ceca, and reduced from the rinse, background microflora. The C. coli strain was orally or intracloacally inoculated into chicks. At 1, 3, and 6 weeks of age, inoculated broilers were removed and several tissue types sampled for the presence of the marker strain. At 6 weeks of age, 10 additional noninoculated penmates were sampled. The C. coli strain colonized chicks, disseminated to body tissues, colonized penmates, and persisted throughout the 6-week grow-out. The C. coli strain's unique characteristic, being resistant to high levels of gentamicin, allows for a marker that can be used in a wide range of Campylobacter research projects.

The purpose of this study was to compare a conventional culture broth method (Bolton enrichment), a newly developed proprietary broth method (TECRA Campylobacter enrichment), and direct plating for recovery of Campylobacter spp. from chicken carcass rinsates. Whole carcass rinses were taken from 140 carcasses at rehang (immediately after defeathering but before evisceration) and from 140 carcasses at postchill from eight different processing plants in the United States. The rinsate samples were packed in ice and shipped overnight to the laboratory. Aliquots of the rinsate were transferred into Bolton and TECRA enrichment broths and were direct plated. Standard laboratory procedures with Campy-cefex plates were followed for recovery of Campylobacter spp. For rehang carcasses, 94% were positive for Campylobacter spp. with the TECRA enrichment broth and 74% were positive with the Bolton enrichment broth. For postchill carcasses, 74% were positive for Campylobacter spp. with the TECRA enrichment broth and 71% were positive with the Bolton enrichment broth. Compared with the Bolton enrichment broth, TECRA enrichment broth significantly suppressed non-Campylobacter microflora (P < 0.05). Overall, TECRA enrichment broth yielded an 11% higher total number of Campylobacter-positive samples compared with the Bolton enrichment broth. Campylobacter spp. detection in postchill samples was significantly greater (P < 0.05) by enrichment (84%) than by direct plating (19%). The high number of Campylobacter-positive samples obtained with all procedures indicated that 99% of the carcass rinsates obtained at rehang and 84% obtained at postchill contained Campylobacter spp.

The ability of the lesser mealworm, Alphitobius diaperinus (Panzer), commonly known as the darkling beetle, to transmit marker Salmonella Typhimurium to day-of-hatch broiler chicks was evaluated, as well as the spread to nonchallenged pen mates. In trial 1, day-of-hatch chicks were orally gavaged with 4 larval or 4 adult beetles that had been exposed to marker Salmonella-inoculated feed for 72 h. In addition, chicks were gavaged with the marker Salmonella in saline solution. These chicks were then placed into pens to serve as challenged broilers. In trial 2, all pens received 2 challenged chicks that were gavaged with larvae or beetles that had been exposed to marker Salmonella-inoculated feed for 24 h and then removed from the inoculated feed for a period of 7 d. At 3 wk of age, cecal samples from the marker Salmonella-challenged broilers and from 5 pen mates in trial 1, or 10 pen mates in trial 2, were evaluated for the presence of the marker Salmonella in their ceca, and at 6 wk of age, all remaining pen mates were sampled. To monitor the presence of the marker Salmonella within pens, stepped-on drag swab litter samples were taken weekly. For the Salmonella-saline pens, 29 to 33% of the broilers that had been challenged and 10 to 55% of the pen mates were positive at 3 wk of age, and only 2 to 6% had positive ceca at 6 wk. For the pens challenged with adult beetles, 0 to 57% of the challenged broilers and 20 to 40% of the pen mates had positive ceca at 3 wk, and 4 to 7% were positive at 6 wk. The pens challenged with larvae had the greatest percentage of marker Salmonella-positive broilers; 25 to 33% of the challenged broilers and 45 to 58% of pen mates were positive at 3 wk, and 11 to 27% were positive at 6 wk. These results demonstrated that ingestion of larval or adult beetles contaminated with a marker Salmonella could be a significant vector for transmission to broilers.

Of the over 200 known extrasolar planets, just 14 pass in front of and behind their parent stars as seen from Earth. This fortuitous geometry allows direct determination of many planetary properties. Previous reports of planetary thermal emission give fluxes that are roughly consistent with predictions based on thermal equilibrium with the planets' received radiation, assuming a Bond albedo of approximately 0.3. Here we report direct detection of thermal emission from the smallest known transiting planet, HD 149026b, that indicates a brightness temperature (an expression of flux) of 2,300 +/- 200 K at 8 microm. The planet's predicted temperature for uniform, spherical, blackbody emission and zero albedo (unprecedented for planets) is 1,741 K. As models with non-zero albedo are cooler, this essentially eliminates uniform blackbody models, and may also require an albedo lower than any measured for a planet, very strong 8 microm emission, strong temporal variability, or a heat source other than stellar radiation. On the other hand, an instantaneous re-emission blackbody model, in which each patch of surface area instantly re-emits all received light, matches the data. This planet is known to be enriched in heavy elements, which may give rise to novel atmospheric properties yet to be investigated.

Of the over 200 known extrasolar planets, 14 exhibit transits in front of their parent stars as seen from Earth. Spectroscopic observations of the transiting planets can probe the physical conditions of their atmospheres. One such technique can be used to derive the planetary spectrum by subtracting the stellar spectrum measured during eclipse (planet hidden behind star) from the combined-light spectrum measured outside eclipse (star + planet). Although several attempts have been made from Earth-based observatories, no spectrum has yet been measured for any of the established extrasolar planets. Here we report a measurement of the infrared spectrum (7.5-13.2 microm) of the transiting extrasolar planet HD 209458b. Our observations reveal a hot thermal continuum for the planetary spectrum, with an approximately constant ratio to the stellar flux over this wavelength range. Superposed on this continuum is a broad emission peak centred near 9.65 microm that we attribute to emission by silicate clouds. We also find a narrow, unidentified emission feature at 7.78 microm. Models of these 'hot Jupiter' planets predict a flux peak near 10 microm, where thermal emission from the deep atmosphere emerges relatively unimpeded by water absorption, but models dominated by water fit the observed spectrum poorly.

Experiments were conducted to compare litter sampling methods for the detection of Salmonella. In experiment 1, chicks were challenged orally with a suspension of naladixic acid-resistant Salmonella and wing banded, and additional nonchallenged chicks were placed into each of 2 challenge pens. Nonchallenged chicks were placed into each nonchallenge pen located adjacent to the challenge pens. At 7, 8, 10, and 11 wk of age the litter was sampled using 4 methods: fecal droppings, litter grab, drag swab, and sock. For the challenge pens, Salmonella-positive samples were detected in 3 of 16 fecal samples, 6 of 16 litter grab samples, 7 of 16 drag swabs samples, and 7 of 16 sock samples. Samples from the nonchallenge pens were Salmonella positive in 2 of 16 litter grab samples, 9 of 16 drag swab samples, and 9 of 16 sock samples. In experiment 2, chicks were challenged with Salmonella, and the litter in the challenge and adjacent nonchallenge pens were sampled at 4, 6, and 8 wk of age with broilers remaining in all pens. For the challenge pens, Salmonella was detected in 10 of 36 fecal samples, 20 of 36 litter grab samples, 14 of 36 drag swab samples, and 26 of 36 sock samples. Samples from the adjacent nonchallenge pens were positive for Salmonella in 6 of 36 fecal droppings samples, 4 of 36 litter grab samples, 7 of 36 drag swab samples, and 19 of 36 sock samples. Sock samples had the highest rates of Salmonella detection. In experiment 3, the litter from a Salmonella-challenged flock was sampled at 7, 8, and 9 wk by socks and drag swabs. In addition, comparisons with drag swabs that were stepped on during sampling were made. Both socks (24 of 36, 67%) and drag swabs that were stepped on (25 of 36, 69%) showed significantly more Salmonella-positive samples than the traditional drag swab method (16 of 36, 44%). Drag swabs that were stepped on had comparable Salmonella detection level to that for socks. Litter sampling methods that incorporate stepping on the sample material while in contact with the litter appear to detect Salmonella in greater incidence than traditional sampling methods of dragging swabs over the litter surface.

The star upsilon Andromedae is orbited by three known planets, the innermost of which has an orbital period of 4.617 days and a mass at least 0.69 that of Jupiter. This planet is close enough to its host star that the radiation it absorbs overwhelms its internal heat losses. Here, we present the 24-micrometer light curve of this system, obtained with the Spitzer Space Telescope. It shows a variation in phase with the orbital motion of the innermost planet, demonstrating that such planets possess distinct hot substellar (day) and cold antistellar (night) faces.

Day-old male broiler breeder chicks were obtained from a commercial hatchery and raised as broilers. For Experiment 1, at 5 wk of age, the broilers were orally inoculated with a 10(6) cfu/ml of a characterized strain of Campylobacter jejuni and a cocktail (three naladixic acid-resistant strains) of Salmonella serovars. One week after inoculation, the birds were euthanatized and defeathered. The abdominal cavity was examined and any unabsorbed yolk material (and remaining yolk stalk) and ceca were aseptically removed for microbiological analyses. For each pooled sample (two birds per pool), an aerobic plate count (APC), an Enterobacteriaceae (ENT) count, and a test for the presence of Campylobacter and Salmonella was performed. For Experiment 2, at 5 wk of age, the broilers were orally inoculated with 10(5) cfu/ml of a characterized strain of Campylobacter jejuni. One week after inoculation, the birds (n = 20) were killed, defeathered, and the yolk stalk, attached yolk, or free-floating yolk and ceca were individually analyzed for presence of Campylobacter. For Experiment 1, the Salmonella-inoculated birds had 2/12 ceca and 0/12 unabsorbed yolk samples positive for Salmonella. The average yolk APC was log10 3.4 cfu/g and the average ENT was log10 1.9 cfu/g. For the Campylobacter-inoculated birds, 12/12 ceca and 9/12 unabsorbed yolk samples were positive for Campylobacter. The average yolk APC was log10 3.5 cfu/g and the average ENT was log10 3.1 cfu/g. For Experiment 2, the inoculated Campylobacter birds had 19/20 ceca, 5/20 free floating yolks, and 19/20 yolk stalks positive. In Experiment 1, the inoculated Campylobacter colonized the ceca in every instance and were present in 75% of the unabsorbed yolks. Alternatively, the inoculated Salmonella were not found in any of the unabsorbed yolks and only rarely in the ceca. In Experiment 2, the inoculated Campylobacter was found in very high numbers in the yolk and internal body samples. Determining to what extent these internal bodies and unabsorbed yolks play in bacterial colonization and contamination of the birds at processing has not been determined. The next step will be to determine the incidence of unabsorbed yolks and presence of Campylobacter and Salmonella in these bodies of commercial broilers at processing.

Campylobacter are known to cause acute bacterial gastroenteritis in humans. Poultry products have been implicated as a significant source of these infections. Six experiments were performed to determine whether Campylobacter could be isolated naturally from the primary and secondary lymphoid organs, liver/gallbladder, and ceca of commercial broiler breeder hens. Broiler breeder hens were acquired from different commercial sources during the early, middle, and late lay cycles. The birds were euthanatized, defeathered, and aseptically opened. To reduce the possibility of cross-contamination between samples, the thymus, spleen, and liver/gallbladder were aseptically removed prior to removal of the ceca. Individual samples were placed in sterile bags, packed on ice, and transported to the laboratory for evaluation. In this study Campylobacter were found in 11 of 43 thymii, eight of 43 spleens, four of 43 liver/gallbladders, and 30 of 43 ceca. Overall, 28 of 53 isolates from the above samples were Campylobacter coli and 25 of 53 isolates were found to be Campylobacter jejuni.

Two studies were conducted to determine whether Campylobacter jejuni could rapidly spread and reside in the internal organs of adult broiler breeder hens. In Study 1, university-housed broiler breeders at 22 wk of age were obtained and placed in individual cages. Each hen was intravaginally inoculated weekly from 23 to 32 wk of age with a characterized strain of C. jejuni. At wk 23, 27, and 32, 4 d postinoculation, the hens were euthanized, defeathered, and aseptically opened. In Study 2, university-housed broiler breeder hens were obtained at 42, 53, and 56 wk of age, placed in individual cages, and inoculated either orally or intravaginally with a characterized strain of C. jejuni. To reduce the possibility of cross-contamination among samples, the thymus, spleen, liver, and gallbladder were aseptically removed, prior to the ceca. In both studies, all samples were individually analyzed. In Study 1, at 23 wk of age, C. jejuni was recovered from 4/7 thymii, 2/7 spleens, 5/7 livers and gallbladders, and 6/7 ceca. At 27 wk of age, C. jejuni was recovered from 1/7 thymii and 1/7 ceca. At 32 wk of age, C. jejuni was recovered from 4/11 thymii, 1/11 livers and gallbladders, and 2/11 ceca. In Study 2, C. jejuni was recovered from 2/6 thymii and 5/6 ceca after oral inoculation and 1/6 spleens, 1/6 livers and gallbladders, and 4/6 ceca after vaginal inoculation of 43-wk-old hens. Campylobacter jejuni was recovered from 2/5 thymii, 3/5 spleens, 3/5 livers and gallbladders, and 2/5 ceca after oral inoculation of 53-wk-old hens and 1/5 thymii and 1/5 livers and gallbladders after vaginal inoculation. Campylobacter jejuni was recovered from 1/4 thymii, 2/4 livers and gallbladders, and 1/4 ceca and was not detected in any vaginally inoculated birds of 57-wk-old hens. This study provides evidence that C. jejuni can reside in the internal organs of broiler breeder hens following oral or intravaginal inoculation.

Unabsorbed yolk sacs are being investigated as a possible reservoir for internal Campylobacter and salmonellae contamination of processed poultry carcasses. However, it is unknown at what frequency that unabsorbed yolk sacs persist at the time of processing of broilers and spent breeders. Seven sets of 100 broiler carcasses (at 6 or 8 wk of age) were obtained from commercial processing plants. In addition, 100 52-wk-old broiler breeder males, 100 102-wk-old Leghorn hens, and 300 8-wk-old Athens-Canadian randombred control (ACRBC) broilers were euthanized, and their abdominal cavities were opened for determination of the presence of unabsorbed yolk sacs. Carcasses with obliterated yolk stalks or stalks with no detectable yolk material were categorized as normal. Those with unabsorbed yolk sacs were further separated into 2 groups: 1) attached by the yolk stalk to the small intestine or 2) unattached within the abdominal cavity. Yolk sacs were further classified by size: 1) small was <2 mm in diameter, 2) medium was 2 to 10 mm, and 3) large was >10 mm. From the 300 commercial broiler carcasses that were 6 wk old, 54% were categorized as normal with no detectable yolk sac, 35% had an unabsorbed yolk sac attached to the yolk stalk, and 12% had unattached yolk sacs. From the 400 commercial broiler carcasses that were 8 wk old, 49% of the carcasses were normal, 31% had attached unabsorbed yolk sacs, and 20% had unattached yolk sacs. From the 100 rooster carcasses sampled, 73% were normal, 8% had attached unabsorbed yolk sacs, and 19% had unattached yolk sacs. From the 100 White Leghorn hen carcasses sampled, 88% were normal, 8% had attached unabsorbed yolk sacs, and 4% were unattached yolk sacs. From the 300 ACRBC carcasses sampled, 76% were normal, 4% had attached unabsorbed yolk sacs, and 20% were unattached yolk sacs. The incidence of unabsorbed yolk sacs in present day commercial broilers appears twice as high as for mature roosters, hens, or ACRBC broilers.

It has been demonstrated that horizontal and vertical transmission of Salmonella and Campylobacter can occur in broiler breeder flocks. The mechanism of this transmission is still unclear. Previously negative broiler breeder flocks have been reported to become positive with Salmonella, Campylobacter, or both after the introduction of "spike" roosters at 45 wk of age. To determine whether the rooster semen is a possible source of transmission to hens for colonization, we evaluated the association of both Salmonella and Campylobacter spp. to segments (head, midpiece, and tail) of individual spermatozoa after artificial inoculation. Salmonella typhimurium, Salmonella heidelberg, and Salmonella montevideo, or Campylobacter jejuni (in 0.85% saline) was added to a freshly collected (by abdominal massage) aliquot of pooled semen from roosters housed in individual cages. The semen and bacteria solutions were incubated 1 h at room temperature. Samples were fixed using Karnosvsky and Zamboni fixatives for 24 h prior to centrifuging and rinsing in 0.1 M cacodylate x HCl buffer. Individual aliquot samples were then subjected to both scanning (JSM-5800) and transmission (JEM-1210) electron microscopy. The scanning electron microscopy showed that Salmonella was associated with all 3 segments (head, midpiece, and tail) of the spermatozoa and apparently equally distributed. Campylobacter was mainly associated with the midpiece and tail segments; few isolates were located on the head segment. The transmission electron microscopy showed apparent attachment of Salmonella and Campylobacter to the spermatozoa.

We previously reported the recovery of Campylobacter (naturally colonized) from the ductus deferens of 5 of 101 broiler breeder roosters, and four of those five positive roosters had previously produced Campylobacter-positive semen samples. Those results prompted further evaluation to determine if inoculation route influenced the prevalence or level of Campylobacter contamination of semen, the digestive tract, or reproductive organs. Individually caged roosters, confirmed to be feces and semen negative for Campylobacter, were challenged with a marker strain of Campylobacter jejuni either orally using 1.0 ml of a diluted cell suspension (log(10)4.3 to 6.0 cells), by dropping 0.1 ml of suspension (log(10)5.3 to 7.0 cells) on the everted phallus immediately after semen collection or by dip coating an ultrasound probe in the diluted cell suspension (log(10)4.3 to 6.0 cells) and then inserting the probe through the vent into the colon. Six days postinoculation, individual feces and semen samples were again collected and cultured for Campylobacter. Seven days postinoculation, roosters were killed, the abdomen aseptically opened to expose the viscera, and one cecum, one testis, and both ductus deferens were collected. The samples were then suspended 1:3 (weight/volume) in Bolton enrichment broth for the culture of Campylobacter. Samples were also directly plated onto Cefex agar to enumerate Campylobacter. Campylobacter was recovered 6 days after challenge from feces in 82% of samples (log(10)4.1 colony-forming units [CFU]/g sample), 85% of semen samples (log(10)2.9 CFU/ml), and on the seventh day postchallenge from 88% of cecal samples (log(10)5.8 CFU/g sample). Campylobacter was not directly isolated from any testis sample but was detected following enrichment from 9% (3/33) of ductus deferens samples. Roosters challenged with Campylobacter orally, on the phallus, or by insertion of a Campylobacter dip-coated ultrasound probe were all readily colonized in the ceca and produced Campylobacter-positive semen and feces on day 6 after challenge. The low prevalence of recovery of Campylobacter from the ductus deferens samples and failure to recover from any testis sample suggests that semen may become Campylobacter positive while traversing the cloaca upon the everted phallus. The production of Campylobacter-positive semen could provide a route in addition to fecal-oral for the horizontal transmission of Campylobacter from the rooster to the reproductive tract of the hen.

The dissemination of Salmonella into various lymphoid-like organs in young broiler chicks after oral and intracloacal inoculation was studied. A three-strain cocktail of Salmonella Typhimurium, Salmonella Montevideo, and Salmonella Enteritidis was administered either orally or intracloacally to day-old chicks. After 1 h, 1 day, or 1 week, the ceca, thymus, liver and gallbladder, spleen, and bursa were sampled for the presence of Salmonella. There was a marked difference in the recovery of Salmonella 1 h postinoculation. Only 6 of 50 samples from orally inoculated chicks were positive compared with 33 of 50 samples from cloacally inoculated samples. In comparison, 24 h and 1 week after inoculation, there was no difference in the number of positive samples between oral or cloacal inoculation. The rapidity of the translocation of the Salmonella from the cloacal inoculum compared to the oral inoculum is likely due to the transient time required for Salmonella to move through the alimentary tract. The method of inoculation did not affect the distribution of serogroups. Of the three serotypes in the composite inoculum, the Salmonella Enteritidis (group D) was recovered only twice in replication 1 and not at all in replication 2. Both the Salmonella Typhimurium (serogroup B) and the Salmonella Montevideo (serogroup C1) were recovered extensively throughout the study.

Campylobacter and Salmonella are known to cause acute bacterial gastroenteritis in humans. Raw poultry products have been implicated as a significant source of these infections. Five trials were conducted to determine whether Campylobacter and Salmonella spp. exist naturally in the mature and immature ovarian follicles of late-life broiler breeder hens. Broiler breeder hens ranging from 60 to 66 wk of age were obtained from four different commercial breeder operations. For each trial, the hens were removed from the commercial operation and held overnight at the University of Georgia processing facility. The hens were euthanized, defeathered, and aseptically opened. To reduce the possibility of cross-contamination between samples, first the mature and immature ovarian follicles, then the ceca, were aseptically removed. Individual samples were placed in sterile bags, packed on ice, and transported to the laboratory for evaluation. Overall, Campylobacter was found in 7 of 55 immature follicles, 12 of 47 mature follicles, and 41 of 55 ceca. Campylobacter was found in at least one of each sample of mature follicles and in ceca in each of the five trials. Salmonella was found in 0 of 55 immature follicles, 1 of 47 mature follicles, and 8 of 55 ceca. In this study, the recovery rate of Salmonella from late-life broiler breeder hen ovarian follicles was relatively low. However, the recovery rate of Campylobacter from the hen ovarian follicles was reasonably high, suggesting that these breeder hens could be infecting fertile hatching eggs. Determining how Campylobacter contaminated these ovarian follicles and how many chicks could be colonized from this source are the next steps in helping to elucidate a better understanding of this ecology and the control of Campylobacter in poultry production.