Shigella sonnei utilises colicins during inter-bacterial competition.

The mammalian colon is one of the most densely populated habitats currently recognised, with 1011-1013 commensal bacteria per gram of colonic contents. Enteric pathogens must compete with the resident intestinal microbiota to cause infection. Among these enteric pathogens are Shigella species which cause approximately 125 million infections annually, of which over 90 % are caused by Shigella flexneri and Shigella sonnei. Shigella sonnei was previously reported to use a Type VI Secretion System (T6SS) to outcompete E. coli and S. flexneri in in vitro and in vivo experiments. S. sonnei strains have also been reported to harbour colicinogenic plasmids, which are an alternative anti-bacterial mechanism that could provide a competitive advantage against the intestinal microbiota. We sought to determine the contribution of both T6SS and colicins to the anti-bacterial killing activity of S. sonnei. We reveal that whilst the T6SS operon is present in S. sonnei, there is evidence of functional degradation of the system through SNPs, indels and IS within key components of the system. We created strains with synthetically inducible T6SS operons but were still unable to demonstrate anti-bacterial activity of the T6SS. We demonstrate that the anti-bacterial activity observed in our in vitro assays was due to colicin activity. We show that S. sonnei no longer displayed anti-bacterial activity against bacteria that were resistant to colicins, and removal of the colicin plasmid from S. sonnei abrogated anti-bacterial activity of S. sonnei. We propose that the anti-bacterial activity demonstrated by colicins may be sufficient for niche competition by S. sonnei within the gastrointestinal environment.

Exploring the experiences of parents of Autistic children when attending the diagnostic imaging department for an X-ray examination.

INTRODUCTION: Autism is a neuro-developmental condition which affects the social-emotional skills, behaviour, language, communication skills and flexibility of thoughts of an individual and their sensory processing. This can result in Autistic service users finding it difficult to navigate current healthcare provision and cope with the unpredictable environment. This paper explores the experiences of parents of Autistic children when attending the diagnostic imaging department for an X-ray examination. METHODS: A cross sectional, mixed methods approach was adopted and the initial phase consisting of an online survey for parents to complete is the subject of this paper. The quantitative data was analysed using descriptive statistics and cross comparison between questions was also completed. Thematic analysis was taken to analyse the data from the two open questions at the end of the survey. RESULTS: The online survey results are presented in this paper under four key themes; waiting times and environment, forms of communication, lack of understanding of staff regarding Autism and preparation for the X-ray examination. CONCLUSION: The overall rating of the parents' experience whilst in the X-ray/diagnostic imaging department was positive, however there are several areas which received low scores which need further attention. These were waiting areas, waiting times, staff development and patient preparation. IMPLICATIONS FOR PRACTICE: The development of more inclusive waiting areas is needed, more effective lines of communication between staff to expedite the patient journey where possible, staff development of both radiographers and also support staff and the review of design of more accessible and inclusive patient information.

Model validation for a knowledge and practices survey towards prevention of soil-transmitted helminth infections in rural villages in Indonesia.

The rate of soil-transmitted helminth (STH) infection is estimated to be around 20% in Indonesia. Health promotion and health education are cost-effective strategies to supplement STH prevention and control programs. Existing studies suggest that quantitative tools for knowledge, attitudes and practices (KAP) are important to monitor effective community-based STH interventions. However, evidence is limited regarding the applicability of such tools. This study aims to identify the socio-demographic predictors for STH-related knowledge and practices and validate the quantitative tools in population use. A cross-sectional study design was conducted among residents of 16 villages in Central Java, Indonesia. Adult and child respondents were interviewed to assess general knowledge and practices in relation to STH. Two mixed effects models identified the significant factors in predicting knowledge and practice scores. The model predicted knowledge and practice scores were compared with the observed scores to validate the quantitative measurements developed in this study. Participants' socio-demographic variables were significant in predicting an individual's STH-related knowledge level and their hand washing and hygiene practices, taking into account household-level variability. Model validation results confirmed that the quantitative measurement tools were suitable for assessing STH associated knowledge and behaviour. The questionnaire developed in this study can be used to support school- and community-based health education interventions to maximize the effect of STH prevention and control programs.

Prison officers' experiences of aggression: implications for sleep and recovery.

BACKGROUND: Prison officers are at high risk of assault that can impair their mental as well as physical health. Such experiences can also disrupt sleep, with negative implications for well-being and job performance. To manage this risk, insight is needed into the mechanisms by which experiencing aggression from prisoners can affect officers' sleep quality. By impairing recovery processes, work-related hypervigilance and rumination might be key factors in this association. AIMS: To examine prison officers' personal experiences of aggression and associations with sleep quality. Also, to consider whether work-related hypervigilance and rumination mediate the relationship between exposure to aggression and sleep. METHODS: We assessed prison officers' experiences of aggression and violence, work-related hypervigilance and rumination via an online survey. The PROMIS was used to measure the quality of sleep. RESULTS: The study sample comprised 1,806 prison officers (86.8% male). A significant relationship was found between the frequency of experiences of aggression at work and the quality of sleep. Work-related hypervigilance and rumination were significantly associated with sleep quality and mediated the relationship between workplace aggression and sleep quality. CONCLUSIONS: Our findings suggest that enhancing the safety climate in prisons might improve officers' quality of sleep that, in turn, could benefit their wellbeing and performance. Implementing individual-level strategies to help prison officers manage hypervigilance and rumination, and therefore facilitate recovery, should also be effective in improving their sleep.

New psychoactive substances, safety and mental health in prison officers.

BACKGROUND: The use of new psychoactive substances (NPS) in UK prisons is believed to have increased substantially. As well as posing a significant threat to prisoners' health, NPS use can trigger violent, unpredictable and aggressive behaviour. Dealing with the direct and indirect effects of NPS therefore has the potential to compromise the physical and psychological safety of prison staff. AIMS: This study investigates prison officers' perceptions of NPS use in their workplace and their risk of exposure. Relationships between NPS exposure, the workplace safety climate and mental health were also examined. METHODS: We assessed prison officers' perceptions of the prevalence of NPS use among prisoners in their workplace, their personal exposure and the safety climate in their institution through an online survey. The General Health Questionnaire-12 measured mental health. Descriptive statistics were used to assess officers' perceptions of NPS use in their workplace and their personal exposure and correlations examined relationships between variables. RESULTS: The sample comprised 1956 prison officers (86% male). Most respondents (85%) highlighted NPS as a serious cause for concern in their institution. Two-thirds (66%) reported being personally exposed to NPS at least sometimes, with 22% being exposed once a day or more. Significant relationships were found between officers' perceived NPS exposure, assessments of safety climate and self-reported mental health. CONCLUSIONS: Our findings highlight the need for urgent action to reduce the use of NPS among prisoners. This is likely to improve the safety climate of UK prisons and the mental health of staff.

Deliberating Performance Targets: Follow-on workshop discussing PM10, NO2, CO, and SO2 air sensor targets.

The use of air sensor technology is increasing worldwide for a variety of applications, however, with significant variability in data quality. The United States Environmental Protection Agency held a workshop in July 2019 to deliberate possible performance targets for air sensors measuring particles with aerodynamic diameters of 10 µm or less (PM10), nitrogen dioxide (NO2), carbon monoxide (CO), and sulfur dioxide (SO2). These performance targets were discussed from the perspective of non-regulatory applications and with the sensors operating primarily in a stationary mode in outdoor environments. Attendees included representatives from multiple levels of government organizations, sensor developers, environmental nonprofits, international organizations, and academia. The workshop addressed the current lack of sensor technology requirements, discussed fit-for-purpose data quality needs, and debated transparency issues. This paper highlights the purpose and key outcomes of the workshop. While more information on performance and applications of sensors is available than in past years, the performance metrics, or parameters used to describe data quality, vary among the studies reports and there is a need for more clear and consistent approaches for evaluating sensor performance. Organizations worldwide are increasingly considering, or are in the process of developing, sensor performance targets and testing protocols. Workshop participants suggested that these new guidelines are highly desirable, would help improve data quality, and would give users more confidence in their data. Given the wide variety of uses for sensors and user backgrounds, as well as varied sensor design features (e.g., communication approaches, data tools, processing/adjustment algorithms and calibration procedures), the need for transparency was a key workshop theme. Suggestions for increasing transparency included documenting and sharing testing and performance data, detailing best practices, and sharing data processing and correction approaches.

A cost-effectiveness analysis of sentinel lymph node detection in vulvar cancer by preoperative lymphoscintigraphy versus intraoperative detection alone.

OBJECTIVE: To determine cost-effectiveness of preoperative lymphoscintigraphy (LSG) for detection of inguinofemoral sentinel lymph nodes (SLN). METHOD: We compared the use of preoperative LSG prior to SLN excision versus omission of preoperative LSG. The two outcomes were death or survival. Costs associated with the procedure were determined by CPT code and published estimates. Cost analysis was performed using Treeage software, and incremental cost-effectiveness ratios (ICERs) were calculated. The measure of effectiveness was incremental survival benefit. ICER thresholds for considering LSG to be cost-effective were based on the value of a statistical life (VSL). RESULTS: Using a baseline probability of 0.93 for finding SLN with LSG, our model estimated LSG costs were $2783.84 with 84.7% survival. Our model then estimated the cost and survival without LSG by varying the SLN detection rate. Survival was equivalent when probability of SLN detection without LSG was 0.93. If detection without LSG was >0.93, not performing LSG was the dominant strategy. Costs were equal when probability of finding SLN without LSG was 0.6. For any SLN detection without LSG below 0.6, performing LSG was the dominant strategy. Formal cost-effectiveness analysis was performed using ICERs for probabilities from 0.60 to 0.93. In this range, costs were higher with LSG, but survival was improved. As long as the incremental detection with LSG was at least 1.05% to 1.47% higher, LSG was cost-effective with ICERs below the VSL. CONCLUSION: In our model, LSG is cost-effective as long as it increases detection of SLN by at least 1.05-1.47%.

Modelling diverse sources of Clostridium difficile in the community: importance of animals, infants and asymptomatic carriers.

Clostridium difficile infections (CDIs) affect patients in hospitals and in the community, but the relative importance of transmission in each setting is unknown. We developed a mathematical model of C. difficile transmission in a hospital and surrounding community that included infants, adults and transmission from animal reservoirs. We assessed the role of these transmission routes in maintaining disease and evaluated the recommended classification system for hospital- and community-acquired CDIs. The reproduction number in the hospital was 1 for nearly all scenarios without transmission from animal reservoirs (range: 1.0-1.34). However, the reproduction number for the human population was 3.5-26.0%) of human exposures originated from animal reservoirs. Symptomatic adults accounted for <10% transmission in the community. Under conservative assumptions, infants accounted for 17% of community transmission. An estimated 33-40% of community-acquired cases were reported but 28-39% of these reported cases were misclassified as hospital-acquired by recommended definitions. Transmission could be plausibly sustained by asymptomatically colonised adults and infants in the community or exposure to animal reservoirs, but not hospital transmission alone. Under-reporting of community-onset cases and systematic misclassification underplays the role of community transmission.

Seasonality and community interventions in a mathematical model of Clostridium difficile transmission.

BACKGROUND: Clostridium difficile infection (CDI) is the leading cause of antibiotic-associated diarrhoea with peak incidence in late winter or early autumn. Although CDI is commonly associated with hospitals, community transmission is important. AIM: To explore potential drivers of CDI seasonality and the effect of community-based interventions to reduce transmission. METHODS: A mechanistic compartmental model of C. difficile transmission in a hospital and surrounding community was used to determine the effect of reducing transmission or antibiotic prescriptions in these settings. The model was extended to allow for seasonal antibiotic prescriptions and seasonal transmission. FINDINGS: Modelling antibiotic seasonality reproduced the seasonality of CDI, including approximate magnitude (13.9-15.1% above annual mean) and timing of peaks (0.7-1.0 months after peak antibiotics). Halving seasonal excess prescriptions reduced the incidence of CDI by 6-18%. Seasonal transmission produced larger seasonal peaks in the prevalence of community colonization (14.8-22.1% above mean) than seasonal antibiotic prescriptions (0.2-1.7% above mean). Reducing transmission from symptomatic or hospitalized patients had little effect on community-acquired CDI, but reducing transmission in the community by ≥7% or transmission from infants by ≥30% eliminated the pathogen. Reducing antibiotic prescription rates led to approximately proportional reductions in infections, but limited reductions in the prevalence of colonization. CONCLUSION: Seasonal variation in antibiotic prescription rates can account for the observed magnitude and timing of C. difficile seasonality. Even complete prevention of transmission from hospitalized patients or symptomatic patients cannot eliminate the pathogen, but interventions to reduce transmission from community residents or infants could have a large impact on both hospital- and community-acquired infections.

Retroperitoneal lymphadenopathy secondary to joint replacement wear and debris, a case report.

We present a case of a 63-year-old female who initially presented with complaints of vulvar swelling and was subsequently found to have isolated retroperitoneal lymphadenopathy on imaging. Biopsy was performed and was indicative of weakly polarizable material that raised the consideration of joint prosthesis wear debris. Due to the patient's concern for an underlying malignancy of gynecologic origin, a laparoscopic lymphadenectomy was ultimately performed and final pathology was consistent with reactive changes due to joint wear and debris from the patient's bilateral total knee arthroplasties placed approximately 16 years prior. While this is a rare presentation, it is important to consider this in the differential of retroperitoneal lymphadenopathy as these patients are often referred to a gynecologic oncologist for further workup.

A persistent mass: A case of aggressive Angiomyxoma of the vulva.

We present a case of aggressive angiomyxoma of the vulva. The patient presented with a persistent, enlarging vulvar mass, initially misdiagnosed as a Bartholin gland cyst. The patient underwent wide local excision, which resulted in total resection of the mass. Final pathology was consistent with aggressive angiomyxoma, a rare soft tissue tumor with a predilection for the female pelvis. Though rare, it is important to consider in the differential diagnosis of a pelvic mass, given the locally aggressive nature of this tumor and propensity for recurrence.

Clostridium difficile classification overestimates hospital-acquired infections.

BACKGROUND: Clostridium difficile infections occur frequently among hospitalized patients, with some infections acquired in hospital and others in the community. International guidelines classify cases as hospital-acquired if symptom onset occurs more than two days after admission. This classification informs surveillance and infection control, but has not been verified by empirical or modelling studies. AIM: To assess current classification of C. difficile acquisition using a simulation model as a reference standard. METHODS: C. difficile transmission was simulated in a range of hospital scenarios. The sensitivity, specificity and precision of classifications that use cut-offs ranging from 0.25 h to 40 days were calculated. The optimal cut-off that correctly estimated the proportion of cases that were hospital acquired and the balanced cut-off that had equal sensitivity and specificity were identified. FINDINGS: The recommended two-day cut-off overestimated the incidence of hospital-acquired cases in all scenarios and by >100% in the base scenario. The two-day cut-off had good sensitivity (96%) but poor specificity (48%) and precision (52%) to identify cases acquired during the current hospitalization. A five-day cut-off was balanced, and a six-day cut-off was optimal in the base scenario. The optimal and balanced cut-offs were more than two days for nearly all scenarios considered (ranges: four to nine days and two to eight days, respectively). CONCLUSION: Current guidelines for classifying C. difficile infections overestimate the proportion of cases acquired in hospital in all model scenarios. To reduce misclassification bias, an infection should be classified as being acquired prior to admission if symptoms begin within five days of admission.

Job demands, resources and mental health in UK prison officers.

BACKGROUND: Research findings indicate that working as a prison officer can be highly stressful, but the aspects of work that predict their mental health status are largely unknown. AIMS: To examine, using elements of the demands-resources model, the extent to which work pressure and several potential resources (i.e. control, support from managers and co-workers, role clarity, effective working relationships and positive change management) predict mental health in a sample of UK prison officers. METHODS: The Health and Safety Executive Management Standards Indicator Tool was used to measure job demands and resources. Mental health was assessed by the General Health Questionnaire-28. The effects of demands and resources on mental health were examined via linear regression analysis with GHQ score as the outcome. RESULTS: The study sample comprised 1267 prison officers (86% male). Seventy-four per cent met 'caseness' criteria for mental health problems. Job demands, poor interpersonal relationships, role ambiguity and, to a lesser extent, low job control and poor management of change were key predictors of mental health status. CONCLUSIONS: The findings of this study can help occupational health practitioners and psychologists develop structured interventions to improve well-being among prison officers.

Hospital effect on infections after four major surgical procedures: outlier and volume-outcome analysis using all-inclusive state data.

BACKGROUND: Hospital volume is known to have a direct impact on the outcomes of major surgical procedures. However, it is unclear if the evidence applies specifically to surgical site infections. AIMS: To determine if there are procedure-specific hospital outliers [with higher surgical site infection rates (SSIRs)] for four major surgical procedures, and to examine if hospital volume is associated with SSIRs in the context of outlier performance in New South Wales (NSW), Australia. METHODS: Adults who underwent one of four surgical procedures (colorectal, joint replacement, spinal and cardiac procedures) at a NSW healthcare facility between 2002 and 2013 were included. The hospital volume for each of the four surgical procedures was categorized into tertiles (low, medium and high). Multi-variable logistic regression models were built to estimate the expected SSIR for each procedure. The expected SSIRs were used to compute indirect standardized SSIRs which were then plotted in funnel plots to identify hospital outliers. FINDINGS: One hospital was identified to be an overall outlier (higher SSIRs for three of the four procedures performed in its facilities), whereas two hospitals were outliers for one specific procedure throughout the entire study period. Low-volume facilities performed the best for colorectal surgery and worst for joint replacement and cardiac surgery. One high-volume facility was an outlier for spinal surgery. CONCLUSIONS: Surgical site infections seem to be mainly a procedure-specific, as opposed to a hospital-specific, phenomenon in NSW. The association between hospital volume and SSIRs differs for different surgical procedures.

Population-level effects of risk factors for bovine respiratory disease in Australian feedlot cattle.

Results obtained from a nationwide longitudinal study were extended to estimate the population-level effects of selected risk factors on the incidence of bovine respiratory disease (BRD) during the first 50days at risk in medium-sized to large Australian feedlots. Population attributable fractions (PAF) and population attributable risks (PAR) were used to rank selected risk factors in order of importance from the perspective of the Australian feedlot industry within two mutually exclusive categories: 'intervention' risk factors had practical strategies that feedlot managers could implement to avoid exposure of cattle to adverse levels of the risk factor and a precise estimate of the population-level effect while 'others' did not. An alternative method was also used to quantify the expected effects of simultaneously preventing exposure to multiple management-related factors whilst not changing exposure to factors that were more difficult to modify. The most important 'intervention' risk factors were shared pen water (PAF: 0.70, 95% credible interval: 0.45-0.83), breed (PAF: 0.67, 95% credible interval: 0.54-0.77), the animal's prior lifetime history of mixing with cattle from other herds (PAF: 0.53, 95% credible interval: 0.30-0.69), timing of the animal's move to the vicinity of the feedlot (PAF: 0.45, 95% credible interval: 0.17-0.68), the presence of Bovine viral diarrhoea virus 1 (BVDV-1) in the animal's cohort (PAF: 0.30, 95% credible interval: 0.04-0.50), the number of study animals in the animal's group 13days before induction (PAF: 0.30, 95% credible interval: 0.10-0.44) and induction weight (PAF: 0.16, 95% credible interval: 0.09-0.23). Other important risk factors identified and prioritised for further research were feedlot region, season of induction and cohort formation patterns. An estimated 82% of BRD incidence was attributable to management-related risk factors, whereby the lowest risk category of a composite management-related variable comprised animals in the lowest risk category of at least four of the five component variables (shared pen water, mixing, move timing, BVDV-1 in the cohort and the number of animals in the animal's group-13). This indicated that widespread adoption of appropriate interventions including ensuring pen water is not shared between pens, optimising animal mixing before induction, timing of the animal's move to the vicinity of the feedlot, and group size prior to placing animals in feedlot pens, and avoiding BVDV-1 in cohorts could markedly reduce the incidence of BRD in medium-sized to large Australian feedlots.

Asymptomatic Clostridium difficile colonization in two Australian tertiary hospitals, 2012-2014: prospective, repeated cross-sectional study.

OBJECTIVES: To investigate the prevalence and risk factors for asymptomatic toxigenic (TCD) and nontoxigenic Clostridium difficile (NTCD) colonization in a broad cross section of the general hospital population over a 3-year period. METHODS: Patients without diarrhoea admitted to two Australian tertiary hospitals were randomly selected through six repeated cross-sectional surveys conducted between 2012 and 2014. Stool specimens were cultured under anaerobic conditions, and C. difficile isolates were tested for the presence of toxin genes and ribotyped. Patients were then grouped into noncolonized, TCD colonized or NTCD colonized for identifying risk factors using multinomial logistic regression models. RESULTS: A total of 1380 asymptomatic patients were enrolled; 76 patients (5.5%) were TCD colonized and 28 (2.0%) were NTCD colonized. There was a decreasing annual trend in TCD colonization, and asymptomatic colonization was more prevalent during the summer than winter months. TCD colonization was associated with gastro-oesophageal reflux disease (relative risk ratio (RRR) = 2.20; 95% confidence interval (CI) 1.17-4.14), higher number of admissions in the previous year (RRR = 1.24; 95% CI 1.10-1.39) and antimicrobial exposure during the current admission (RRR = 2.78; 95% CI 1.23-6.28). NTCD colonization was associated with chronic obstructive pulmonary disease (RRR = 3.88; 95% CI 1.66-9.07) and chronic kidney failure (RRR = 5.78; 95% CI 2.29-14.59). Forty-eight different ribotypes were identified, with 014/020 (n = 23), 018 (n = 10) and 056 (n = 6) being the most commonly isolated. CONCLUSIONS: Risk factors differ between patients with asymptomatic colonization by toxigenic and nontoxigenic strains. Given that morbidity is largely driven by toxigenic strains, this novel finding has important implications for disease control and prevention.

Effects of supplementing methionine hydroxy analog on beef cow performance, milk production, reproduction, and preweaning calf performance.

Mature Simmental × Angus cows (214 cows; 635 ± 7 kg) were utilized to determine the effects of late gestation and early postpartum supplementation of methionine hydroxy analog (MHA) on cow BW, BCS, milk production, milk composition, reproduction, and calf performance until weaning in a fall-calving, cool-season grazing system. Cows were stratified by BW, age, AI sire, and assigned to 1 of 12 pastures (17 or 18 cows·pasture). Pastures were randomly allotted to 1 of 2 treatments: control (0.45 kg·cow·d of wheat midd-based pellets, = 6) or supplement including MHA (0.45 kg·cow·d of wheat midd-based pellets including 10 g MHA supplied as MFP (Novus International, Inc., St. Charles, MO; = 6). Treatments were fed 23 ± 7 d prepartum through 73 ± 7 d postpartum. Cow BW was collected at postcalving (27 ± 7 d postpartum), end of supplementation (73 ± 7 d postpartum), AI, pregnancy check, and end of trial (192 and 193 ± 7 d postpartum). At 73 ± 7 d postpartum, a subset of cow-calf pairs was used in a weigh-suckle-weigh procedure to determine milk production, and milk samples were taken to determine milk composition ( = 45·treatment). Serum from blood was collected at 73 ± 7 and 83 ± 7 d postpartum to determine cow cyclicity and concentrations of 2-hydroxy4-(methylthio) butanoic acid (HMTBa) and L-Methionine. After supplementation, all cow-calf pairs were managed as a common group until weaning (193 ± 7 d of age). Cows were bred via AI at 97 ± 7 d postpartum and clean-up bulls were turned out 11 d post-AI for a 55-d breeding season. Cows fed MHA had greater ( < 0.01) serum concentrations of HMTBa. Cow BW and BCS were not different ( ≥ 0.10) at any time points between treatments. There was no treatment effect ( ≥ 0.17) on calf birth BW, calf weaning BW (193 ± 7 d of age), or calf ADG. Calculated 24-h milk production, milk composition and component production did not differ ( ≥ 0.21). There were no differences ( ≥ 0.50) in percentage of cows cycling, AI conception rate, and overall pregnancy rate between treatments. Post-trial nutritional modeling suggests cows experienced several nutritional deficiencies beyond methionine (Met) that limited the response to Met supplementation. Although supplementation of MHA during late gestation through estimated peak lactation increased serum HMTBa concentrations, it did not affect cow performance, cow milk production, or calf performance when fall-calving cows grazed cool-season forages.

Quantifying accessibility and use of improved sanitation: towards a comprehensive indicator of the need for sanitation interventions.

To prevent diseases associated with inadequate sanitation and poor hygiene, people needing latrines and behavioural interventions must be identified. We compared two indicators that could be used to identify those people. Indicator 1 of household latrine coverage was a simple Yes/No response to the question "Does your household have a latrine?" Indicator 2 was more comprehensive, combining questions about defecation behaviour with observations of latrine conditions. Using a standardized procedure and questionnaire, trained research assistants collected data from 6,599 residents of 16 rural villages in Indonesia. Indicator 1 identified 30.3% as not having a household latrine, while Indicator 2 identified 56.0% as using unimproved sanitation. Indicator 2 thus identified an additional 1,710 people who were missed by Indicator 1. Those 1,710 people were of lower socioeconomic status (p < 0.001), and a smaller percentage practiced appropriate hand-washing (p < 0.02). These results show how a good indicator of need for sanitation and hygiene interventions can combine evidences of both access and use, from self-reports and objective observation. Such an indicator can inform decisions about sanitation-related interventions and about scaling deworming programmes up or down. Further, a comprehensive and locally relevant indicator allows improved targeting to those most in need of a hygiene-behaviour intervention.

Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods.

In this study, we have evaluated the efficacy of propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) to differentiate between viable and non-viable Ancylostoma caninum ova. The newly developed method was validated using raw wastewater seeded with known numbers of A. caninum ova. Results of this study confirmed that PMA-qPCR has resulted in average of 88 % reduction (P < 0.05) in gene copy numbers for 50 % viable +50 % non-viable when compared with 100 % viable ova. A reduction of 100 % in gene copies was observed for 100 % non-viable ova when compared with 100 % viable ova. Similar reductions (79-80 %) in gene copies were observed for A. caninum ova-seeded raw wastewater samples (n = 18) collected from wastewater treatment plants (WWTPs) A and B. The newly developed PMA-qPCR method was applied to determine the viable ova of different helminths (A. caninum, A. duodenale, Necator americanus and Ascaris lumbricoides) in raw wastewater, human fecal and soil samples. None of the unseeded wastewater samples were positive for the above-mentioned helminths. N. americanus and A. lumbricoides ova were found in unseeded human fecal and soil samples. For the unseeded human fecal samples (1 g), an average gene copy concentration obtained from qPCR and PMA-qPCR was found to be similar (6.8 × 10(5) ± 6.4 × 10(5) and 6.3 × 10(5) ± 4.7 × 10(5)) indicating the presence of viable N. americanus ova. Among the 24 unseeded soil samples tested, only one was positive for A. lumbricoides. The mean gene copy concentration in the positively identified soil sample was 1.0 × 10(5) ± 1.5 × 10(4) (determined by qPCR) compared to 4.9 × 10(4) ± 3.7 × 10(3) (determined by PMA-qPCR). The newly developed PMA-qPCR methods were able to detect viable helminth ova from wastewater and soil samples and could be adapted for health risk assessment.