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The microbiome is composed of many organisms and is impacted by an intricate exchange between genetics and environmental factors. The perinatal microbiome influences both the developing fetus and the pregnant person. The purpose of this article is to describe the tests that are currently available for laboratory analysis of the perinatal microbiome in relationship to probiotic interventions. This article focuses on the bacterial component of the microbiome. Although adverse outcomes associated with the perinatal microbiome have been studied, a comprehensive understanding of the physiologic perinatal microbiome is still emerging. Early efforts to influence the perinatal microbiome through probiotics are currently under investigation. Unique terminology is defined, and the microbial composition of perinatal microbiota is summarized. The outcomes of studies of antenatal probiotics are summarized. Microbiome testing and analysis are defined and compared. Implications for perinatal care and probiotics research are presented.

Objective:Healthcare-associated infections (HAIs) are a significant burden on healthcare facilities. Universal gloving is a horizontal intervention to prevent transmission of pathogens that cause HAI. In this meta-analysis, we aimed to identify whether implementation of universal gloving is associated with decreased incidence of HAI in clinical settings.Methods:A systematic literature search was conducted to find all relevant publications using search terms for universal gloving and HAIs. Pooled incidence rate ratios (IRRs) and 95% confidence intervals (CIs) were calculated using random effects models. Heterogeneity was evaluated using the Woolf test and the I2 test.Results:In total, 8 studies were included. These studies were moderately to substantially heterogeneous (I2 = 59%) and had varied results. Stratified analyses showed a nonsignificant association between universal gloving and incidence of methicillin-resistant Staphylococcus aureus (MRSA; pooled IRR, 0.94; 95% CI, 0.79–1.11) and vancomycin-resistant enterococci (VRE; pooled IRR, 0.94; 95% CI, 0.69–1.28). Studies that implemented universal gloving alone showed a significant association with decreased incidence of HAI (IRR, 0.77; 95% CI, 0.67–0.89), but studies implementing universal gloving as part of intervention bundles showed no significant association with incidence of HAI (IRR, 0.95; 95% CI, 0.86–1.05).Conclusions:Universal gloving may be associated with a small protective effect against HAI. Despite limited data, universal gloving may be considered in high-risk settings, such as pediatric intensive care units. Further research should be performed to determine the effects of universal gloving on a broader range of pathogens, including gram-negative pathogens.

We studied farmworker practices and beliefs potentially contributing to transmission of bacteria and their associated antibiotic resistance genes (ARGs) among animals and farm workers to identify potential behavioral interventions to reduce the risk of bacterial transmission. Ten focus groups were conducted on eight Wisconsin dairy farms to assess potentially high-risk practices and farmworker knowledge and experiences with antibiotic use and resistance using the Systems Engineering in Patient Safety (SEIPS) framework. Farmworkers were asked to describe common on-farm tasks and the policies guiding these practices. We found workers demonstrated knowledge of the role of antibiotic stewardship in preventing the spread of ARGs. Worker knowledge of various forms of personal protective equipment was higher for workers who commonly reported glove-use. Additionally, workers knowledge regarding the importance of reducing ARG transmission varied but was higher than we had hypothesized. Programs to reduce ARG spread on dairy farms should focus on proper hand hygiene and personal protective equipment use at the level of knowledge, beliefs, and practices.

We studied farmworker practices and beliefs potentially contributing to transmission of bacteria and their associated antibiotic resistance genes (ARGs) among animals and farm workers to identify potential behavioral interventions to reduce the risk of bacterial transmission. Ten focus groups were conducted on eight Wisconsin dairy farms to assess potentially high-risk practices and farmworker knowledge and experiences with antibiotic use and resistance using the Systems Engineering in Patient Safety (SEIPS) framework. Farmworkers were asked to describe common on-farm tasks and the policies guiding these practices. We found workers demonstrated knowledge of the role of antibiotic stewardship in preventing the spread of ARGs. Worker knowledge of various forms of personal protective equipment was higher for workers who commonly reported glove-use. Additionally, workers knowledge regarding the importance of reducing ARG transmission varied but was higher than we had hypothesized. Programs to reduce ARG spread on dairy farms should focus on proper hand hygiene and personal protective equipment use at the level of knowledge, beliefs, and practices.

Use of antimicrobials in animal agriculture is under increasing scrutiny, but the quantity of antimicrobials used on large US dairy farms has not been evaluated using data from large farms and different metrics. This study investigated total antimicrobial usage (AMU) in adult dairy cows and preweaned calves (PWC) and contrasted 2 metrics used for measurement of AMU. Wisconsin dairy farms were eligible if they had >250 lactating cows, maintained computerized animal health records, and were willing to allow researchers access to treatment records. Animal health data for a 1-yr period was retrospectively collected from computerized records, and a farm visit was performed to verify case definitions and recording accuracy. Both dose-based (animal daily doses; ADD) and mass-based (total mg of antimicrobials per kg of body weight; BW) metrics were calculated at the herd, cow, and PWC levels. Descriptive statistics for AMU were examined for both age groups. Mean AMU was compared among active ingredients and route of usage using ANOVA models that included farm as a random variable. At enrollment, farms (n = 40) contained approximately 52,639 cows (mean: 1,316 ± 169; 95% CI: 975, 1657) and 6,281 PWC (mean: 180 ± 33; 95% CI: 112, 247). When estimated using ADD, total herd AMU was 17.2 ADD per 1,000 animal-days (95% CI: 14.9, 19.5), with 83% of total herd-level AMU in adult cows. When estimated using the mass-based metric, total herd AMU was 13.6 mg of antimicrobial per kilogram of animal BW (95% CI: 10.3, 17.0), with 86% of total AMU used in adult cows. For cows, 78% of total ADD (15.8 ADD per 1,000 cow-d) was administered as intramammary (IMM) preparations. In contrast, when AMU was estimated using a mass-based metric, IMM preparations represented only 24% of total AMU (12.1 mg of antimicrobial/kg of cow BW). For cows, ceftiofur was the primary antimicrobial used and accounted for 53% of total ADD, with 80% attributed to IMM and 20% attributed to injectable treatments. When estimated using a mass-based metric, ampicillin was the predominant antimicrobial used in cows and accounted for 33% of total antimicrobial mass per kilogram of BW. When AMU was estimated for PWC using ADD, injectable antimicrobials represented 79% of total usage (28.3 ADD per 1,000 PWC-d). In contrast, when AMU was estimated for PWC using a mass-based metric, injectable products represented 42% of total AMU, even though more farms administered antimicrobials using this route. When AMU in PWC was summarized using ADD, penicillin represented 32% of AMU, and there were no significant differences in ADD among ampicillin, oxytetracycline or enrofloxacin. When a mass-based metric was used to estimate AMU in PWC, oral products (sulfadimethoxine and trimethoprim-sulfa) represented more than half of the total AMU given to this group. Overall, these results showed that choice of metric and inclusion of different age groups can substantially influence interpretation of AMU on dairy farms.

Public health agencies have recommended that the public wear face coverings as a major non-pharmaceutical intervention to mitigate COVID-19 transmission. However, there is concern whether or not the public has adopted this recommendation. An observational study of 3,271 members of the public was conducted during the COVID-19 pandemic and examined the use of face coverings at grocery stores across 20 Wisconsin counties between May 16th and June 1st, 2020. Of the total individuals observed, we found that only 41.2% used face coverings. Individuals who appeared to be adults (aOR = 1.65; 95% CI = 1.20-2.27) or older adults (aOR = 3.67; 95% CI = 2.59-5.19) were more likely to be wearing face coverings than younger individuals. Additionally, individuals with female gender expression (aOR = 1.43; 95% CI = 1.24-1.66) and individuals shopping at a more expensive grocery store (aOR = 1.95; 95% CI = 1.69-2.25) were more likely to be wearing face coverings. We did not find an association between county level prevalence of COVID-19 cases and face covering use. To our knowledge, this is the first study using direct observations to examine face covering behavior by the public and our findings have implications for public health agencies.

We examined the effect of chlorhexidine gluconate (CHG) bathing on the skin microbiota of adult and pediatric patients. We observed no differences in pediatric patients; however, multiple genera of bacteria were observed to be significantly less abundant in the adults bathing with CHG. Further research is needed to determine the long-term impact of CHG use on the skin microbiota.

Background: Up to 30% of patients with Clostridioides difficile infection (CDI) develop recurrent infection, which is associated with a 33% increased risk of mortality at 180 days. The gut microbiome plays a key role in initial and recurrent episodes of CDI. We examined the clinical characteristics and gut microbial diversity in patients with recurrent (rCDI) versus nonrecurrent CDI at a tertiary-care academic medical center. Methods: Stool samples were collected from 113 patients diagnosed with CDI between 2018 and 2019. Clinical and demographic data were extracted from the electronic medical record (Table 1), and 16S rRNA sequencing of the v4 region was carried out on the Illumina MiSeq using 2×250 paired-end reads. Sequences were binned into operational taxonomic units (OTUs) using mothur and were classified to the genus level whenever possible using the ribosomal database project data set version 16. Alpha diversity was calculated using the Shannon diversity index. Β diversity was calculated using the Bray-Curtis dissimilarity matrix. Differential abundance testing was done using DESeq to assess taxonomic differences between groups. A P value of .05 was used to assess significance. Results: In total, 55 patients had rCDI (prior positive C. difficile polymerase chain reaction in last 7–365 days) and 58 had nonrecurrent CDI (Table 1). Patients with rCDI had a higher frequency of organ transplant and comorbidity. No differences in α not β diversity were observed between groups. Also, 4 OTUs were more abundant in those with rCDI: Ruminococcus (n = 2), Odoribacter, and Lactobacillus. Patients with rCDI had microbiomes with greater proportions of Bacteroidetes (27% of OTUs) compared to the nonrecurrent group (18%) as well as fewer OTUs belonging to the Firmicutes phyla compared to the nonrecurrent patients (56% vs 59%). Among the rCDI patients, those experiencing 2 or more recurrences had greater abundances of Bacteroides and Ruminococcus, while those experiencing only 1 recurrence had significantly greater abundances of Akkermensia, Ruminococcus, Streptococcus, Roseburia, Clostridium IV, and Collinsella compared to those with only 1 recurrence (Table 2). Conclusions: Patients with rCDI had a more impaired microbiome than those with initial CDI. Ruminococcus OTUs have been previously indicated as a risk factor for recurrence and treatment failure, and they were significantly more abundant in those with rCDI and among those with multiple recurrences. The greatest differences in the microbiome were observed between those with 1 recurrence compared to those with multiple recurrences. Interventions for gut microbiome restoration should focus particularly on those with recurrent CDI.Funding: NoDisclosures: None

Background To determine the prevalence of intestinal S. aureus colonization of patients at a large teaching hospital and determine the molecular characteristics of the identified strains. The second objective of this research was to determine risk factors associated with S. aureus intestinal colonization. Methods A cross-sectional study of 781 specimens from inpatients and outpatients at the University of Iowa Hospitals and Clinics Clinical Microbiology Laboratory was conducted. S. aureus was identified using traditional culture methodologies. Methicillin-resistance was determined via PCR of the mecA gene. PVL PCR, spa typing, and antimicrobial sensitivity testing were also done. A nested case-control study was done on a subset of patients with all colonized patients defined as cases and non-colonized controls. Medical record abstractions were done to identify risk factors for intestinal colonization in the nested study. Results Out of 625 patients included in the final study, 58 were positive for S. aureus (9.3%). One isolate was positive for the PVL gene. A high number of isolates were resistant to multiple antibiotics including oxacillin (43.1%), erythromycin (51.7%), and levofloxacin (41.4%). All isolates were susceptible to vancomycin, daptomycin, linezolid, and quinupristin-dalfopristin. In the nested study, having a disease or condition of the gastrointestinal tract significantly increased the odds of intestinal colonization (OR: 1.96, 95% CI: 1.04–3.7; aOR: 13.9, 95% CI: 1.67–115.7). No other variables were significantly associated with increased odds of colonization. Conclusions S. aureus was identified from the stool of patients at the University of Iowa Hospitals and Clinics, with a large number of those isolates being resistant to antibiotics and may serve a reservoir for subsequent infections as well as asymptomatic transmission.

Supplemental Digital Content is available in the text., Background: Infection by antibiotic resistant bacteria (ARB) is a global health crisis and asymptomatic colonization increases risk of infection. Nonhuman studies have linked heavy metal exposure to the selection of ARB; however, few epidemiologic studies have examined this relationship. This study analyzes the association between urinary lead level and colonization by ARB in a nonclinical human population. Methods: Data came from the Survey of the Health of Wisconsin 2016–2017, and its ancillary Wisconsin Microbiome Study. Urinary lead levels, adjusted for creatinine, were used to assess exposure. ARB included methicillin resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), fluoroquinolone resistant Gram-negative bacilli (RGNB), and Clostridium difficile (C. diff), from skin, nose, and mouth swabs, and saliva and stool samples. Logistic regression, adjusted for covariates, was used to evaluate associations between Pb and ARB. Secondary analysis investigated Pb resistance from ARB isolates. Results: Among 695 participants, 239 (34%) tested positive for ARB. Geometric mean urinary Pb (unadjusted) was 0.286 µg/L (95% confidence intervals [CI] = 0.263, 0.312) for ARB negative participants and 0.323 µg/L (95% CI = 0.287, 0.363) for ARB positive participants. Models adjusted for demographics, diet, and antibiotic use showed elevated odds of positive colonization for those in the 95th percentile (vs. below) of Pb exposure (odds ratio [OR] = 2.05, 95% CI = 0.95, 4.44), and associations were highest in urban residents (OR = 2.85, 95% CI = 1.07, 7.59). RGNB isolates were most resistant to Pb. Discussion: These novel results suggest that Pb exposure is associated with increased colonization by ARB, and that RGNB are particularly resistant to Pb.

The role of household meat handling and consumption in the transfer of Staphylococcus aureus (S. aureus) from livestock to consumers is not well understood. Examining the similarity of S. aureus colonizing humans and S. aureus in meat from the stores in which those individuals shop can provide insight into the role of meat in human S. aureus colonization. S. aureus isolates were collected from individuals in rural and urban communities in Iowa (n = 3347) and contemporaneously from meat products in stores where participants report purchasing meat (n = 913). The staphylococcal protein A (spa) gene was sequenced for all isolates to determine a spa type. Morisita indices and Permutational Multivariate Analysis of Variance Using Distance Matrices (PERMANOVA) were used to determine the relationship between spa type composition among human samples and meat samples. spa type composition was significantly different between households and meat sampled from their associated grocery stores. spa types found in meat were not significantly different regardless of the store or county in which they were sampled. spa types in people also exhibit high similarity regardless of residential location in urban or rural counties. Such findings suggest meat is not an important source of S. aureus colonization in shoppers.

Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of infectious disease morbidity and mortality. Previous studies have confirmed the presence of S. aureus, including MRSA, on raw meat products. We investigated the prevalence and molecular epidemiology of S. aureus and MRSA in commercially-distributed antibiotic-free and conventional raw meat products (n = 3290) purchased in 8 Iowa retail stores weekly for a period of one year. Isolates were characterized using spa typing, and PCR was used to detect the presence of the Panton-Valentine leukocidin (PVL) and mecA genes. Quantitation of S. aureus on meat products was carried out one week per month. The prevalence of S. aureus on meat samples was 27.8% (913/3290). Compared to antibiotic-free meat samples, higher prevalence of both MRSA and methicillin-susceptible S. aureus (MSSA) were found in conventional meat samples. Among the S. aureus isolates, 18 were PVL-positive (1.9%) and 41 (4.5%) carried mecA. Phenotypic oxacillin resistance was observed for 17.1% (41/239) of the isolates tested, while 23% (55/239) were multi-drug resistant. A total of 132 spa types were detected from 913 contaminated meat samples. Overall, t002 was the most common spa type identified (137; 15.0%). The number of colony-forming units (CFU) per 10 g meat ranged from 2 to 517 (median: 8 CFU per 10 g of meat; mean: 28) with the highest bacterial load observed on turkey samples. These data reinforce the need to consider meat products as potential vehicles of S. aureus transmission from farm into human households, and the potential need for public health intervention programs pre and post-slaughter in meat processing facilities.

Background: Antimicrobials are used on dairy farms for preventing disease and treating common infections such as mastitis. Objective: We aimed to understand farmworker practices that potentially contribute to transmission of antimicrobial resistance bacteria and their genes (ARG) among animals and farm workers, and to identify human behavioral interventions to reduce risk. Methods: Focus groups with farm workers were held at 8 dairy farms across Wisconsin selected to represent a range of antibiotic use in cattle. We explored the nature of potentially high-risk practices and farm-worker knowledge and experiences with antibiotic use and resistance. Farm workers were asked to describe common tasks, including hand hygiene and eating practices, and the policies guiding these practices. Focus groups were conducted in English and Spanish guided by the Systems Engineering in Patient Safety (SEIPS) framework, adapted for an agricultural context. Discussions were recorded, transcribed, and translated. A content analysis was conducted to identify themes. Dedoose version 8.0.35 software was used to organize the data. Results: In total, 10 focus groups were conducted on 8 farms. Knowledge of when to use antibiotics for human health varied; upset stomach, headache, and flu symptoms were suggested as appropriate uses. Few workers had personal experience with antibiotic resistance at home or on the farm. Some displayed knowledge of the role of antibiotic stewardship in preventing the spread of ARG (“I guess all dairy farmers have a responsibility not to overdo it”). Others associated the risk of spread with the consumption of raw milk or meat from cows receiving antibiotics. Knowledge of personal protective equipment was stronger among workers who commonly reported glove use. Some perceived glove use to be mandatory, and others chose to wear gloves in the perceived absence of written rules. Some workers reported changing gloves numerous times throughout the day, and others did so less frequently or “only when they rip.” In general, hand hygiene practices are guided by individual knowledge of established rules, beliefs about risk, and personal discretion. Conclusions: Knowledge about mechanisms of spread of ARGs varies among workers on Wisconsin dairy farms and reflects a combination of farm-level rules, experience, individual knowledge, and beliefs. Applying knowledge from the healthcare setting to reduce ARG spread into agriculture is crucial to the tenets of One Health. Programs to reduce ARG spread on dairy farms should focus on proper hand hygiene and PPE use at the level of knowledge, beliefs, and practices.Funding: Funding: was provided by the USDA-NIFA Food Safety Challenge (grant no. 2017-68003-26500).Disclosures: None

Background: Children attending daycare are at increased risk of carrying multidrug-resistant organisms (MDROs) compared to children not attending daycare. Carriage of MDROs greatly increases the risk of infection, not only in the child but also for others living in the household. Understanding the epidemiology of MDRO carriage in children is essential to devising effective containment strategies. Here, we present the findings from a cross-sectional study assessing MDRO carriage in daycare-attending and nonattending children in Wisconsin. Methods: We applied the following enrollment criteria: Children aged between 6 months and H. influenzae, S. pneumoniae, M. catarrhalis, and methicillin-resistant S. aureus (MRSA). Skin samples were cultured for MRSA, and stool samples were cultured for MRSA, C. difficile, vancomycin-resistant enterococci (VRE), and extended-spectrum β-lactamase–producing Gram-negative bacilli (ie, ESBL GNR). Results: In total, 44 children were enrolled in this study. The average age was 2.6 years and 50% were girls. Furthermore, 30 (68.2%) were identified by their parents as white, 9 (20.5%) as black, and 5 (11.3%) as other or multiracial. Incidentally, 23 children (52.3%) were enrolled in daycare. Overall, 18 children were positive for at least 1 organism, 9 of which had daycare exposure, and 5 children (1 in daycare) were positive for >1 organism (11.4%). From stool samples, 6 children (13.6%, 2 in daycare) were C. difficile carriers, 3 were VRE carriers (6.8%, 1 in daycare), 8 carried an ESBL GNR (18.2%, 4 in daycare), and 3 carried MRSA (6.8%, 1 in daycare). One child was positive for H. influenzae (2.3%, not in daycare) and 2 were positive for S. pneumoniae (4.6%, 1 in daycare) from nares swabs. One child was positive for MRSA (2.3%, not in daycare) from a skin swab. We detected no significant differences between children with and without daycare exposure for any organism. Conclusions: Children in this population had higher than expected rates of ESBL GNRs and MRSA for a community population. Daycare exposure was not correlated with increased carriage in this small pilot study, though larger longitudinal studies are needed.Funding: NoneDisclosures: None

Recurrent Clostridiodes difficile infections (rCDIs) are a burdensome problem. Patients with a history of CDI that are prescribed antibiotics are at a high risk for recurrence. Fecal microbiota transplantation (FMT) has been shown to be an effective treatment for rCDI, though there is little information on the impact of FMT with antibiotics on the gut microbiome. We are conducting a clinical trial of FMT to prevent rCDI in patients with a history of CDI currently taking antibiotics. Our primary objective is to determine the effect of FMT on the gut microbiome during antibiotic exposure. Our secondary aim is to assess safety and feasibility of using FMT as a prophylaxis for CDI. We plan to enroll 30 patients into a phase II randomized, double-blind, placebo-controlled trial with three arms: (1) 5 FMT capsules per day during antibiotic treatment and for 7 days post antibiotic cessation, (2) a one-time dose of 30 FMT capsules 48–72 h post cessation of antibiotic treatment, or (3) 5 placebo capsules per day during antibiotic treatment and for 7 days post antibiotic treatment. Patients provide stool samples throughout the duration of the study and are cultured C. difficile. Sequencing of the V4 region of the 16S rRNA gene will be carried out to assess the gut microbiota. Results of this study will provide information on the impact of FMT on the gut microbiome as well as the necessary data to examine whether or not prophylactic FMT should be explored further as a way to prevent CDI recurrence.

IntroductionApproximately 25%–35% of the 1991 Gulf War Veteran population report symptoms consistent with Gulf War Illness (GWI), a chronic, multi-symptom illness characterised by fatigue, pain, irritable bowel syndrome and problems with cognitive function. GWI is a disabling problem for Gulf War Veterans, and there remains a critical need to identify innovative, novel therapies.Gut microbiota perturbation plays a key role in the symptomatology of other chronic multi-symptom illnesses, including myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). Given similarities between ME/CFS and GWI and the presence of gastrointestinal disorders in GWI patients, Veterans with GWI may also have gut abnormalities like those seen with ME/CFS. In this longitudinal cohort study, we are comparing the diversity (structure) and the metagenomes (function) of the gut microbiome between Gulf War Veterans with and without GWI. If we find differences in Veterans with GWI, the microbiome could be a target for therapeutic intervention to alleviate GWI symptoms.Methods and analysisParticipants answer questions about diet, exercise and lifestyle factors. Participants also complete a questionnaire (based on the Kansas case definition of GWI) regarding their medical history and symptoms; we use this questionnaire to group participants into GWI versus healthy control cohorts. We plan to enrol 52 deployed Gulf War Veterans: 26 with GWI and 26 healthy controls. Participants provide stool and saliva samples weekly for an 8-week period for microbiome analyses. Participants also provide blood samples at the beginning and end of this period, which we will use to compare measures of inflammation markers between the groups.Ethics and disseminationThe protocol was approved by the University of Wisconsin-Madison Health Sciences Institutional Review Board and the William S. Middleton Memorial Veterans Hospital Research and Development Committee. Results of this study will be submitted for publication in a peer-reviewed journal.

Probiotics may represent a promising approach for reducing Clostridioides (Clostridium) difficile infections (CDIs). A clinical trial conducted by our group demonstrated that CDI patients undergoing adjunctive treatment with Lactobacillus and Bifidobacterium probiotics had a reduction in diarrheal duration and compositional changes in their stool microbiomes. Here, we modified a CDI mouse model to represent clinical outcomes observed in patients and employed this model to identify evidence for the prevention of primary CDI and relapse with the same probiotic. Mice (n = 80) were administered 0.25 mg/ml cefoperazone over 5 days and subsequently challenged with 10(2) C. difficile VPI 10463 spores. A subset of mice (n = 40) were administered 10(8) CFU of probiotics daily alongside cefoperazone pretreatment and until experimental endpoints were reached. Clinical scoring was performed daily on mice and used to evaluate CDI onset and severity. Moderate CDI in mice was defined by survival beyond day 3 postinfection, while mice with severe CDI were those who succumbed to infection prior to day 3 postinfection. Sequencing and analysis of 16S rRNA from stool content were performed to determine compositional alterations to the microbiota. Using total clinical scores, we identified an association between probiotic treatment and delayed onset of primary CDI and relapse by approximately 12 to 24 h (P

Little information exists on the microbiomes of livestock workers. A cross-sectional, epidemiological study was conducted enrolling 59 participants (26 of which had livestock contact) in Iowa. Participants were enrolled in one of four ways: from an existing prospective cohort study (n=38), from the Iowa Department of Natural Resources Animal Feeding Operations database (n=17), through Iowa county fairs (n=3), and through snowball sampling (n=1). We collected two sets of swabs from the nares and oropharynx of each participant. The first set of swabs was used to assess the microbiome via 16s rRNA sequencing and the second was used to cultureS. aureus.We observed livestock workers to have greater diversity in their microbiomes compared to those with no livestock contact. In the nares, there were 26 operational taxonomic units found to be different between livestock workers and non-livestock workers with the greatest difference seen withStreptococcusandProteobacteria.In the oropharynx, livestock workers with swine exposure were more likely to carry several pathogenic organisms. The results of this study are the first to characterize the livestock worker nasal and oropharyngeal microbiomes.

Imbalance of the gut microbiota has been linked to many adverse health outcomes including infection, and autoimmune disease. Along with its other detrimental health effects, lead (Pb) exposure may ...

The study objective was to determine the prevalence ofStaphylococcus aureuscolonisation in the nares and oropharynx of healthy persons and identify any risk factors associated with suchS. aureuscolonisation. In total 263 participants (177 adults and 86 minors) comprising 95 families were enrolled in a year-long prospective cohort study from one urban and one rural county in eastern Iowa, USA, through local newspaper advertisements and email lists and through the Keokuk Rural Health Study. Potential risk factors including demographic factors, medical history, farming and healthcare exposure were assessed. Among the participants, 25.4% of adults and 36.1% minors carriedS. aureusin their nares and 37.9% of adults carried it in their oropharynx. The overall prevalence was 44.1% among adults and 36.1% for minors. Having at least one positive environmental site forS. aureusin the family home was associated with colonisation (prevalence ratio: 1.34, 95% CI: 1.07–1.66). The sensitivity of the oropharyngeal cultures was greater than that of the nares cultures (86.1% compared with 58.2%, respectively). In conclusion, the nares and oropharynx are both important colonisation sites for healthy community members and the presence ofS. aureusin the home environment is associated with an increased probability of colonisation.

Background Infectious agents have the potential to thrive in child daycare facilities. Asymptomatic Staphylococcus aureus carriage is a risk factor for developing infection and contributes to transmission. Methods We collected swabs from 110 employees, 111 unexposed adults, 81 children, and 214 environmental surfaces at 11 Iowa daycare facilities. S aureus isolates were characterized using antibiotic resistance profiles and Staphylococcal protein A typing. Staphylococcal protein A types were grouped into cluster complexes using the Based Upon Repeat Pattern algorithm. Results All isolates (from 38 employees, 37 unexposed adults, 16 children, and 19 surfaces) were characterized. Daycare employees were more likely to carry erythromycin-resistant S aureus than unexposed adults (odds ratio, 3.7; 95% confidence interval, 1.1-12.7; P = .033). Isolates were genetically heterogeneous, although isolates from employees appeared more clonal than those from unexposed adults. Strains associated with ST8 were identified in 5 daycare facilities and 3 unexposed adults. Conclusions S aureus isolates collected from employees, children, and surfaces of daycare facilities are genetically heterogeneous, but contain strains associated with community-associated methicillin-resistant S aureus . This suggests that daycare facilities can serve as reservoirs for community-associated methicillin-resistant S aureus and facilitate genetic exchange. Employees may be at increased risk of carrying antibiotic-resistant strains, indicating more research is necessary into this occupational group.

Examine the relationship between colonization with Staphylococcus aureus in the community and symptomatic infection in two cohorts of Iowans.Case series within cohort study.Rural Iowans selected from the Keokuk Rural Health Study, the Agricultural Health Study, and the Iowa Voter Registry.Longitudinal study within established cohorts evaluating documented S. aureus infections with samples available for molecular typing.We examined this relationship in two cohorts of Iowans with a combined 11 incident cases of S. aureus SSTI, for which samples were available. Seven of the 11 individuals (63.6%) were colonized at baseline, in the nose (3/7, 42.9%), or in both the nose and throat (57.1%). All seven cases had matching sequence types between colonization and infection isolates.Staphylococcus aureus causes millions of skin and soft tissue infections yearly. Although colonization with S. aureus is a frequent antecedent to infection, many studies investigating the link between colonization and infection have taken place in a clinical setting, particularly in urban hospitals. Our study has shown similar results in a rural community setting to those previously seen in clinics.

Perturbations in the gastrointestinal microbiome caused by antibiotics are a major risk factor for Clostridium difficile infection (CDI). Probiotics are often recommended to mitigate CDI symptoms; however, there exists only limited evidence showing probiotic efficacy for CDI. Here, we examined changes to the GI microbiota in a study population where probiotic treatment was associated with significantly reduced duration of CDI diarrhea. Subjects being treated with standard of care antibiotics for a primary episode of CDI were randomized to probiotic treatment or placebo for 4 weeks. Probiotic treatment consisted of a daily multi-strain capsule (Lactobacillus acidophilus NCFM, ATCC 700396; Lactobacillus paracasei Lpc-37, ATCC SD5275; Bifidobacterium lactis Bi-07, ATCC SC5220; Bifidobacterium lactis B1-04, ATCC SD5219) containing 1.7 x 1010 CFUs. Stool was collected and analyzed using 16S rRNA sequencing. Microbiome analysis revealed apparent taxonomic differences between treatments and timepoints. Subjects administered probiotics had reduced Verrucomicrobiaceae at week 8 compared to controls. Bacteroides were significantly reduced between weeks 0 to 4 in probiotic treated subjects. Ruminococcus (family Lachnospiraceae), tended to be more abundant at week 8 than week 4 within the placebo group and at week 8 than week 0 within the probiotic group. Similar to these results, previous studies have associated these taxa with probiotic use and with mitigation of CDI symptoms. Compositional prediction of microbial community function revealed that subjects in the placebo group had microbiomes enriched with the iron complex transport system, while probiotic treated subjects had microbiomes enriched with the antibiotic transport system. Results indicate that probiotic use may impact the microbiome function in the face of a CDI; yet, more sensitive methods with higher resolution are warranted to better elucidate the roles associated with these changes. Continuing studies are needed to better understand probiotic effects on microbiome structure and function and the resulting impacts on CDI.

BackgroundStaphylococcus aureusis a frequent cause of both infections globally. Colonization with the organism is known to increase the risk of developing infections and occurs in roughly one third of the general population. While many factors influence colonization, it has been demonstrated other members of the microbiome influence colonization withS. aureus. Here, we assessed the nasal and oropharyngeal microbiomes of healthy participants in relation toS. aureuscolonization in a cross-sectional study using 16s rRNA sequencing of the v1-v3 region. As livestock workers have also been shown to be at an increased risk of carriage, we have also assessed microbiota differences in colonization status in a population of livestock workers.ResultsIn both the nares and oropharynx, there were no microbiota differentially abundant between colonized and non-colonized persons. However, there was a significant difference in the beta diversity (Bray-Curtis distances) between carriers and non-carriers (P=0.002). When considering carriage stratified by livestock exposure, there were a number of differences. Most notably, colonized livestock workers had significantly morePorphyomonas(2-fold change = -8.54,P= 0.03) than the non-colonized livestock workers.ConclusionsS. aureusis a frequent colonizer of the human upper respiratory tract, including the nares and oropharynx and causes a wide range of infections. Livestock workers are at increased risk for carriage. Interventions such as improving oral hygiene may lead to decreasedS. aureuscarriage by reducing other bacterial species such asPorphyomonas. Larger, longitudinal studies are needed to better explore what microorganisms may be associated withS. aureuscolonization.

BackgroundS. aureusis a frequent cause of hospital and community associated infections and colonization is known to increase the risk of infection, with the nares considered the most important colonization site.MethodsWe compared the prevalence of nasal and oropharyngeal carriage in a yearlong, prospective cohort study of people from the community as well as assessed risk factors for nares-only and oropharynx-only colonization.ResultsColonization at both anatomical sites was correlated; however, oropharynx only carriage occurred and oropharyngeal swabs were more sensitive than nasal swabs at detecting carriage (77.27% and 72.725 respectively). Non-Caucasian race, having a greater number of people living in your home and more children in your home all significantly increased the odds of oropharynx-only carriage. HavingS. aureuspresent on home environmental sites, exercising in a fitness center, and sharing bath towels all increased the odds of nares-only carriage.ConclusionsOropharyngeal swabs increase the detection ofS. aureuscolonization in community embers.

Summary: Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a devastating pathogen that is associated with high morbidity and mortality worldwide. Livestock are a well-known reservoir for this pathogen, which poses substantial health risks for livestock workers. Little is known about the epidemiology of livestock-associated MRSA (LA-MRSA) among livestock workers in Eastern Europe. Methods: To study the epidemiology of LA-MRSA among swine workers in Romania, we collected and characterized nasal and oropharygneal samples from swine workers on commercial pig farms. A survey that included questions about work-related tasks, biosafety practices, contact with animals, and health status was used to assess the risk factors that were potentially associated with LA-MRSA colonization. Results: The prevalence of MRSA colonization among swine workers was 6.8%. Two LA-MRSA strains with the spa types t034 and t011 and one likely community-associated MRSA strain with the spa type t321 were isolated from workers on five farms. Interestingly, all MRSA carriers worked on farms that imported animals from other production facilities. Conclusion: This is the first study to confirm the presence of LA-MRSA among swine workers in Romania and suggests the need to minimize the risk of LA-MRSA-related infections in swine workers and their community contacts. The findings also suggest a link between the commercial movement of swine and the introduction of LA-MRSA. Keywords: Methicillin-resistant Staphylococcus aureus (MRSA), Antimicrobial resistance, Occupational exposure, Zoonoses, Swine

Background Little is known about the relationship between oral hygiene and multidrug-resistant organism in the mouth and gut. We aimed to assess the relationship of oral hygiene and diet with multidrug-resistant organism (MDRO) carriage in the oral cavity and gut. Methods Participants were adults over age 18 from the 2016–2017 Survey of the Health of Wisconsin (SHOW) and its ancillary Wisconsin Microbiome Study. SHOW surveys residents of Wisconsin, collecting health determinants including a food frequency questionnaire, oral health, as well as biologic specimens. MDROs were defined as the presence of methicillin-resistant Staphylococcus aureus, Vancomycin-resistant Enterococcus, and Fluoroquinolone-resistant Gram-negative bacteria identified via culture from saliva, oral swabs, and stool samples. Statistical analysis was performed in R v3.5.1. Univariate analyses were conducted for all variables in the data set. Any variable with a P-value < 0.2 in the univariate analysis was considered for the logistic regression. Logistic regression using the glm function was done modeling MDRO carriage in either the saliva, oral swab, saliva and oral samples combined, and stool against diet, oral health, and known confounders. Results 876 participants were included in the dataset with all 876 providing oral and stool samples and 784 providing saliva samples. Thirty-three patients were MDRO positive in the saliva (4.2%), 36 were positive in the oral swabs (4.1%), 55 were positive in either the saliva or oral swabs (6.3%), and 103 were positive in the stool (11.8%). In the logistic regression, consumption of whole grains was significantly associated with reduced MDRO carriage in the saliva (P = 0.046) and saliva and oral swab combined (P = 0.036) data sets (Table 1). Conclusion Consuming more whole grains was associated with a lower prevalence of MDRO carriage in the oral cavity. Higher levels of sugar consumption were associated with a higher prevalence of MDRO in the gut. Oral hygiene was not found to be associated with MDRO colonization in the mouth and a higher prevalence in the gut in this cross-sectional study. This may be due to over-reporting of hygiene practices by participants. Being positive for an MDRO in the oral cavity significantly increased the risk of MDRO carriage in the gut. Disclosures All authors: No reported disclosures.

Background: Lead (Pb) is a ubiquitous environmental contaminant with an array of detrimental health effects in children and adults, including neurological and immune dysfunction. Emerging evidence suggests that Pb exposure may alter the composition of the gut microbiota, however few studies have examined this association in human populations. The purpose of this study was to examine the association between urinary Pb concentration and the composition of the adult gut microbiota in a population-based sample of adults. Methods: Data used in this study were collected as part of the Survey of the Health of Wisconsin (SHOW) and its ancillary microbiome study. The SHOW is a household-based health examination survey of Wisconsin residents, collecting a variety of survey data on health determinants and outcomes, as well as objective measurements of body habitus, and biological specimens including urine. The ancillary microbiome study added additional questions and biological specimen collection, including stool, from participants age 18+. Pb concentration was analyzed in urine samples, and gut microbiota composition was assessed using DNA sequencing of the 16S rRNA V4 region, extracted from stool samples. Data processing and statistical analyses were performed in mothur, Python, R, and SAS. Results: Of 696 participants, urinary Pb concentration was highest in those age 70+, females, those with a high school diploma or lower, current and former smokers, and those without indoor pets. In adjusted models, increasing urinary Pb levels were associated with increases in microbial α-diversity (p = 0.071) and richness (p = 0.005). Differences in microbial β-diversity were significantly associated (p = 0.003) with differences in urinary Pb level. Presence of Proteobacteria, including members of the Burkholderiales, was significantly associated with increased urinary Pb. Conclusion: These results suggest that Pb exposure is associated with differences in the composition of the adult gut microbiota in a population-based human sample. Further investigation of this association is warranted. Keywords: Heavy metals, Lead, Microbiome, Microbiota, 16S rRNA, Epidemiology

IntroductionPrevention of multidrug-resistant organism (MDRO) infections, such as those caused by methicillin-resistantStaphylococcus aureus, vancomycin-resistant enterococci, fluoroquinolone-resistant Gram-negative bacteria andClostridium difficileis crucial. Evidence suggests that dietary fibre increases gut microbial diversity, which may help prevent colonisation and subsequent infection by MDROs. The aim of the Winning the War on Antibiotic Resistance (WARRIOR) project is to examine associations of dietary fibre consumption with the composition of the gut microbiota and gut colonisation by MDROs. The secondary purpose of the study is to create a biorepository of multiple body site specimens for future microbiota research.Methods and analysisThe WARRIOR project collects biological specimens, including nasal, oral and skin swabs and saliva and stool samples, along with extensive data on diet and MDRO risk factors, as an ancillary study of the Survey of the Health of Wisconsin (SHOW). The SHOW is a population-based health survey collecting data on several different health determinants and outcomes, as well as objective body measurements and biological specimens. WARRIOR participants include 600 randomly selected Wisconsin residents age 18 and over. Specimens are screened for MDRO colonisation and DNA is extracted for 16S ribosomal RNA-based microbiota sequencing. Data will be analysed to assess the relationship between dietary fibre, the gut microbiota composition and gut MDRO colonisation.Ethics and disseminationThe WARRIOR project is approved by the University of Wisconsin Institutional Review Board. The main results of this study will be published in a peer-reviewed scientific journal.

Swine and their products have become a central part of food systems around the world. Global pork production has rapidly increased over the past 30 years, leading to the intensification of the swine industry: though there are fewer farms now, those farms that do persist raise ever-larger numbers of animals. This increases the transmission of pathogens both amongst animal herds, and between animals and their human caretakers. Furthermore, increased stress to animals and the potential for amplification of pathogens in the farming environment can lead to a higher burden of disease-causing organisms in and on meat products, which then make their way to consumers world-wide. As such, swine and their meat products have the potential to introduce new zoonotic diseases into populations via multiple routes of transmission. Here we discuss several examples of zoonotic diseases of swine origin, reviewing diseases with bacterial, viral, or parasitic causes.

Staphylococcus aureus colonizes approximately 30% of Americans and causes a range of infections [1]. An estimated 1.5% of the US population is colonized with methicillin-resistant S. aureus (MRSA) [2, 3]; emergence of multidrug-resistant strains increases this burden [4]. Staphylococcus aureus epidemiology has changed considerably in recent decades. Livestock-associated S. aureus (LA-SA) strains, including sequence type (ST) 398 and ST9, are often associated with swine, cattle, and poultry exposure [5], but little is known regarding LA-SA carriage and infection in the United States [6, 7]. ST398 carriage has been reported in areas with a high density of swine [8–12]. ST398 is the dominant MRSA strain on European farms [6, 13] and has been documented in the United States [11, 14]. It is unknown if ST398 colonizes a broader population of livestock workers or if additional strains may be zoonotic and spreading on US farms. Common human strains of t002/ST5 and t008/ST8 have been found in live pigs in North America [15–19]. Invasive human disease caused by ST398 has been reported, even in those without animal contact, suggesting that bidirectional spread of these strains is possible [6]. Many studies have investigated S. aureus colonization and infection in the healthcare setting [20]. Fewer have assessed its ecology and transmission in the community [21], and most of these have been conducted in urban areas [22, 23]. In a rural state like Iowa, which ranks third nationally in overall livestock production and first in swine production, transmission of S. aureus between pigs and farmers and into the broader community could complicate efforts to control S. aureus transmission statewide, and could have effects nationally due to movement of colonized pigs or travel by colonized humans. In this study, we report the prevalence of S. aureus colonization and infection among a population of Iowans, along with molecular characteristics, antimicrobial resistance profiles, and risk factors for carriage. We identified potential transmission within households and associations between duration of livestock exposure and LA-SA carriage.