The effectiveness of treating irrigation water using ultraviolet radiation or sulphuric acid fertilizer for reducing generic Escherichia coli on fresh produce-a controlled intervention trial.

AIMS: The aims of this study were to: (i) estimate the effectiveness of ultraviolet radiation (UV) and sulphuric acid-based fertilizer (SA), at reducing levels of generic Escherichia coli in surface irrigation water and on produce and surface soil in open produce fields; and (ii) describe the population dynamics of generic E. coli in produce fields. METHODS AND RESULTS: Spinach and cantaloupe plots were randomly assigned to control, UV or SA treatment groups. Irrigation water was inoculated with Rifampicin-resistant E. coli prior to treatment. More than 75% of UV- and SA-treated tank water samples had counts below the detection limit, compared to a mean count of 3·3 Log10 CFU per ml before treatment. Levels of Rifampicin-resistant E. coli in soil and produce both increased and decreased over 10-15 days after irrigation, depending on the plot and time-period. CONCLUSIONS: UV and SA treatments effectively reduce the levels of E. coli in surface irrigation water. Their effectiveness at reducing contamination on produce was dependent on environmental conditions. Applying wait-times after irrigation and prior to harvest is not a reliable means of mitigating against contaminated produce. SIGNIFICANCE AND IMPACT OF THE STUDY: The results are of timely importance for the agricultural industry as new FSMA guidelines require producers to demonstrate a low microbial load in irrigation water or allow producers to apply a wait-time to mitigate the risk of contaminated produce.

Public perceptions of antibiotic use on dairy farms in the United States.

There has been a global push for improved antimicrobial stewardship, including in animal agriculture, due to growing concerns about antimicrobial resistance. However, little is known about the general public's perceptions of antimicrobial use in animal agriculture. The aim of this study was to explore the US public's perceptions of antibiotic use in dairy farming and how these perceptions influence purchasing decisions. Data from the 2017 Cornell National Social Survey developed in collaboration with the Cornell Survey Research Institute were used to assess the public's perceptions. The Survey Research Institute of Cornell University (Ithaca, NY) administered the survey by telephone to a random sample of 1,000 adults in the continental United States. The survey collected information about perceptions of threat to human health posed by antibiotic use in cows on dairy farms and willingness to pay more for milk from cows raised without antibiotics, as well as several presumed explanatory variables, including respondents' knowledge of antibiotics, beliefs regarding cattle treatment in dairy farming, and 18 sociodemographic characteristics. Data were analyzed using logistic regression. Among respondents, 90.7% (n = 892/983) reported that antibiotic use on dairy farms posed some level of threat to human health and 71.5% (n = 580/811) indicated they would be willing to pay more for milk produced from cows raised without antibiotics. Respondents who believed that antibiotic use in dairy farming posed a moderate to high threat to human health were more likely to be female and report willingness to pay more for milk or not purchase milk. Additionally, consumers' willingness to pay more for milk from cattle raised without antibiotics was associated with the belief that antibiotic use posed some threat to human health, the belief that cows are treated better on organic dairy farms, an annual household income of $50,000 or greater, being born outside the United States, having a liberal social ideology, and being currently or formerly married. These results suggest that the general public's decisions as consumers of dairy products are associated with demographic factors in addition to perceptions of antibiotic use and cattle treatment in dairy farming. The rationale behind such perceptions should be further explored to facilitate consumers' informed decision making about antibiotic use in agriculture, links to cattle treatment, and associated willingness-to-pay attitudes.

Interobserver Agreement Using Histological Scoring of the Canine Liver.

BACKGROUND: Grading schemes for the assessment of hepatic fibrosis and necroinflammatory activity in humans previously have been applied to dogs with chronic hepatitis. Interobserver agreement is a desirable characteristic for any histological scoring scheme. HYPOTHESIS/OBJECTIVES: To assess interobserver agreement associated with pathologists using a previously published histological scoring scheme to assess hepatic fibrosis and necroinflammatory activity in dogs and to compare fibrosis scores assigned to serial sections stained with hematoxylin & eosin (H&E) and picrosirius red. ANIMALS: Histological sections of liver from 50 dogs with variable degrees of hepatic fibrosis and necroinflammatory activity were selected from institutional tissue archives. METHODS: Six board-certified veterinary anatomic pathologists assigned fibrosis and necroinflammatory activity scores to the histological sections. The multiuser kappa statistic was calculated to assess interobserver agreement. Fibrosis stage assigned to serial sections stained with picrosirius red and H&E was compared using the Wilcoxon signed-rank test. RESULTS: Multiuser kappa statistics for assessment of fibrosis and necroinflammatory activity from H&E-stained sections were 0.35 and 0.16, respectively. There was no difference in median fibrosis scores assigned to serial section stained with H&E and picrosirius red (P = .248). CONCLUSIONS AND CLINICAL IMPORTANCE: There was fair interobserver agreement when pathologists assessed fibrosis and poor agreement when they assessed necroinflammatory activity. This suboptimal agreement must be taken into account by clinicians making decisions based on histology reports of the liver and in the design of studies evaluating these findings. To decrease this variability, ideally >1 pathologist should evaluate each section.

Assessment of microbial risk factors and impact of meteorological conditions during production of baby spinach in the Southeast of Spain.

There is a timely need to evaluate the effect agricultural factors and meteorological conditions on fresh produce contamination. This study evaluated those risk factors and described, for the first time, the distribution of indicator microorganisms (Escherichia coli, Enterococcus, coliforms, and Enterobacteriaceae) and the prevalence of foodborne pathogens (Enterohaemorrhagic E. coli, Listeria monocytogenes and Salmonella spp.) in baby spinach grown in the Southeast of Spain. A longitudinal study was conducted on three farms (2011-2013). Results obtained for E. coli highlighted soil and irrigation water as important factors affecting the microbial safety of baby spinach. Significant differences in the proportion of E. coli positive samples were found between treated (46.1%) and untreated (100%) irrigation water. However, the microbial quality of irrigation water didn't affect E. coli prevalence in produce. All E. coli positive spinach samples were detected at the highest observed temperature range suggesting that ambient temperature affects the probability and extent of spinach contamination. Salmonella spp. was detected by RT-PCR in manure, soil, irrigation water and baby spinach but only two of them (manure and irrigation water) were confirmed by isolation in culture media. Salmonella RT-PCR positive samples showed higher levels of E. coli than Salmonella negative samples. This preliminary finding supports recent identification of E. coli as a suitable parameter for the hygiene criterion at the primary production of leafy greens.

Cost-benefit analysis of avian influenza control in Nepal.

Numerous outbreaks of highly pathogenic avian influenza A strain H5N1 have occurred in Nepal since 2009 despite implementation of a national programme to control the disease through surveillance and culling of infected poultry flocks. The objective of the study was to use cost-benefit analysis to compare the current control programme (CCP) with the possible alternatives of: i) no intervention (i.e., absence of control measures [ACM]) and ii) vaccinating 60% of the national poultry flock twice a year. In terms of the benefit-cost ratio, findings indicate a return of US $1.94 for every dollar spent in the CCP compared with ACM. The net present value of the CCP versus ACM, i.e., the amount of money saved by implementing the CCP rather than ACM, is US $861,507 (the benefits of CCP [prevented losses which would have occurred under ACM] minus the cost of CCP). The vaccination programme yields a return of US $2.32 for every dollar spent when compared with the CCR The net present value of vaccination versus the CCP is approximately US $12 million. Sensitivity analysis indicated thatthe findings were robust to different rates of discounting, whereas results were sensitive to the assumed market loss and the number of birds affected in the outbreaks under the ACM and vaccination options. Overall, the findings of the study indicate that the CCP is economically superior to ACM, but that vaccination could give greater economic returns and may be a better control strategy. Future research should be directed towards evaluating the financial feasibility and social acceptability of the CCP and of vaccination, with an emphasis on evaluating market reaction to the presence of H5N1 infection in the country.

Estimating the probability of an extinction or major outbreak for an environmentally transmitted infectious disease.

Indirect transmission through the environment, pathogen shedding by infectious hosts, replication of free-living pathogens within the environment, and environmental decontamination are suspected to play important roles in the spread and control of environmentally transmitted infectious diseases. To account for these factors, the classic Susceptible-Infectious-Recovered-Susceptible epidemic model is modified to include a compartment representing the amount of free-living pathogen within the environment. The model accounts for host demography, direct and indirect transmission, replication of free-living pathogens in the environment, and removal of free-living pathogens by natural death or environmental decontamination. Based on the assumptions of the deterministic model, a continuous-time Markov chain model is developed. An estimate for the probability of disease extinction or a major outbreak is obtained by approximating the Markov chain with a multitype branching process. Numerical simulations illustrate important differences between the deterministic and stochastic counterparts, relevant for outbreak prevention, that depend on indirect transmission, pathogen shedding by infectious hosts, replication of free-living pathogens, and environmental decontamination. The probability of a major outbreak is computed for salmonellosis in a herd of dairy cattle as well as cholera in a human population. An explicit expression for the probability of disease extinction or a major outbreak in terms of the model parameters is obtained for systems with no direct transmission or replication of free-living pathogens.

Transmission of Escherichia coli O157:H7 in cattle is influenced by the level of environmental contamination.

A pen infection-transmission experiment was conducted to elucidate the role of pathogen strain and environmental contamination in transmission of Escherichia coli O157:H7 (ECO157) in cattle. Five steers were inoculated with a three-strain mixture of ECO157 and joined with five susceptible steers in each of two experimental replicates. Faecal and environmental samples were monitored for ECO157 presence over 30 days. One ECO157 strain did not spread. Transmission rates for the other two strains were estimated using a generalized linear model developed based on a modified 'Susceptible-Infectious-Susceptible' mathematical model. Transmission rates estimated for the two strains (0·11 and 0·14) were similar. However, the rates significantly (P = 0·0006) increased 1·5 times for every 1-unit increase in the level of environmental contamination measured as log10 c.f.u. Depending on the level of environmental contamination, the estimated basic reproduction numbers varied from <1 to 8. The findings indicate the importance of on-farm measures to reduce environmental contamination for ECO157 control in cattle that should be validated under field conditions.

Cross-sectional serosurvey of avian influenza antibodies presence in domestic ducks of Kathmandu, Nepal.

Kathmandu, Nepal has been classified as a high-risk area for highly pathogenic avian influenza (HPAI) by the Nepali Government. While ducks have an important role in the transmission of avian influenza viruses (AIV), including HPAI, seroprevalence of antibodies to AIV in domestic ducks of Kathmandu has never been assessed. The objectives of this study were (i) to estimate the prevalence of seroconversion to AIV in domestic ducks in major duck-raising areas of Kathmandu and (ii) to assess the effect of age, sex, presence of swine and the number of ducks on the farm on the carriage of antibodies to AIV in these ducks. From April through July of 2011, a cross-sectional study was conducted and a total of 310 ducks in the major duck-raising areas of Kathmandu were sampled. The estimated prevalence of AIV antibodies was 27.2% [95% confidence interval (CI): 24.6-29.5]. Of 62 enrolled farms, 42% had at least one seropositive duck. Half of the enrolled farms also kept pigs of which 52% had at least one seropositive duck. Bivariate analysis indicated association between ducks' seroconversion to AIV and their age, sex and farm size. However, the final multivariable model, after controlling for clustering of ducks within farms, identified age as the only significant risk factor. Based on this model, ducks older than 1 year of age were more likely to be seropositive compared to ducks <6 months of age [odds ratio = 2.17 (1.07-4.39)]. These results provide baseline information about the AIV seroprevalence in domestic ducks in the major duck-raising areas of Kathmandu and identify a high-risk group that can be targeted in surveillance activities. Future studies should be conducted to differentiate the subtypes of AIV present among domestic ducks in Kathmandu, with particular interest in the presence of HPAI viruses.

Understanding the role of cleaning in the control of Salmonella Typhimurium in grower-finisher pigs: a modelling approach.

Salmonella Typhimurium (STM) infection in pigs represents a considerable food safety concern. This study used mathematical modelling to evaluate the effectiveness of cleaning (faeces removal) as a measure to control STM spread among grower-finisher pigs. A modified Susceptible-Infected-Recovered-Susceptible (SIRS) model of STM transmission through a contaminated environment was developed. Infected pigs were divided into three states according to the pathogen level being shed in their faeces. Infection transmission was evaluated using the basic reproduction number (R 0) and the prevalence of infectious pigs at slaughter age. Although increased frequency and efficiency of cleaning did reduce the prevalence of STM shedding at the time of slaughter, these efforts alone were not capable of eliminating the infection from the population. The level of STM faecal shedding by infectious pigs strongly influenced the infection spread and prevalence at slaughter. To control STM in pigs, cleaning should be combined with vaccination and/or isolation of high-level shedders.

Identifying areas of high risk of human exposure to coccidioidomycosis in Texas using serology data from dogs.

Coccidioidomycosis or Valley Fever (VF) is an emerging soil-borne fungal zoonosis affecting humans and animals. Most non-human cases of VF are found in dogs, which we hypothesize may serve as sentinels for estimating the human exposure risk. The objective of this study is to use the spatial and temporal distribution and clusters of dogs seropositive for VF to define the geographic area in Texas where VF is endemic, and thus presents a higher risk of exposure to humans. The included specimens were seropositive dogs tested at a major diagnostic laboratory between 1999 and 2009. Data were aggregated by zip code and smoothed by empirical Bayesian estimation to develop an isopleth map of VF seropositive rates using kriging. Clusters of seropositive dogs were identified using the spatial scan test. Both the isopleth map and the scan test identified an area with a high rate of VF-seropositive dogs in the western and southwestern parts of Texas (relative risk = 31). This location overlapped an area that was previously identified as a potential endemic region based on human surveys. Together, these data suggest that dogs may serve as sentinels for estimating the risk of human exposure to VF.

The strain-specific dynamics of Escherichia coli O157:H7 faecal shedding in cattle post inoculation.

This study reports analysis of faecal shedding dynamics in cattle for three Escherichia coli O157:H7 (ECO157) strains (S1, S2 and S3) of different genotype and ecological history, using experimental inoculation data. The three strains were compared for their shedding frequency and level of ECO157 in faeces. A multistate Markov chain model was used to compare shedding patterns of S1 and S2. Strains S1 and S2 were detected seven to eight times more often and at 10(4) larger levels than strain S3. Strains S1 and S2 had similar frequencies and levels of shedding. However, the total time spent in the shedding state during colonization was on average four times longer for S1 (15 days) compared to S2 (4 days). These results indicate that an ECO157 strain effect on the frequency, level, pattern and the duration of faecal shedding may need to be considered in control of ECO157 in the cattle reservoir.

Effectiveness of environmental decontamination as an infection control measure.

The effectiveness of environmental decontamination (ED) as a measure in the control of infectious diseases is controversial. This work quantifies the effectiveness of ED by analysing the transmission of pathogens from the environment to susceptible hosts in a Susceptible-Infected-Susceptible model. Analysis of the model shows that ED can render a population disease-free only when the duration of infection (D) is within a certain range. As host-to-host transmission rate is increased, D falls outside this range and the higher levels of ED have a diminishing return in reducing the number of infected hosts at endemic equilibrium. To avoid this, ED can be combined with other control measures, such as treating infected individuals to push the duration of infection into the specified range. We propose decision criteria and minimum ED efforts required for control policies to be effective.

Modeling of spatially referenced environmental and meteorological factors influencing the probability of Listeria species isolation from natural environments.

Many pathogens have the ability to survive and multiply in abiotic environments, representing a possible reservoir and source of human and animal exposure. Our objective was to develop a methodological framework to study spatially explicit environmental and meteorological factors affecting the probability of pathogen isolation from a location. Isolation of Listeria spp. from the natural environment was used as a model system. Logistic regression and classification tree methods were applied, and their predictive performances were compared. Analyses revealed that precipitation and occurrence of alternating freezing and thawing temperatures prior to sample collection, loam soil, water storage to a soil depth of 50 cm, slope gradient, and cardinal direction to the north are key predictors for isolation of Listeria spp. from a spatial location. Different combinations of factors affected the probability of isolation of Listeria spp. from the soil, vegetation, and water layers of a location, indicating that the three layers represent different ecological niches for Listeria spp. The predictive power of classification trees was comparable to that of logistic regression. However, the former were easier to interpret, making them more appealing for field applications. Our study demonstrates how the analysis of a pathogen's spatial distribution improves understanding of the predictors of the pathogen's presence in a particular location and could be used to propose novel control strategies to reduce human and animal environmental exposure.

Modeling on-farm Escherichia coli O157:H7 population dynamics.

Escherichia coli O157:H7 is a potentially fatal foodborne pathogen with a putative reservoir for human infection in feedlot cattle. In order to more effectively identify targets for intervention strategies, we aimed to (1) assess the role of various feedlot habitats in E. coli O157:H7 propagation and (2) provide a framework for examining the relative contributions of animals and the surrounding environment to observed pathogen dynamics. To meet these goals we developed a mathematical model based on an ecological metapopulation framework to track bacterial population dynamics inside and outside the host. We used E. coli O157:H7 microbiological and epidemiological literature to characterize E. coli O157:H7 habitats at the pen level and account for E. coli O157:H7 population processes in water troughs, feedbunks, cattle hosts, and pen floors in the model. Simulations indicated that E. coli O157:H7 was capable of maintaining viable populations in the feedlot without net growth in the cattle gastrointestinal tract, suggesting E. coli O157:H7 may not always act as an obligate parasite. Water troughs and contaminated pen floors appeared to be particularly influential sources driving E. coli O157:H7 population dynamics and thus would serve as prime environmental targets for interventions to effectively reduce the E. coli O157:H7 load at the pen level.

The effect of heterogeneous infectious period and contagiousness on the dynamics of Salmonella transmission in dairy cattle.

The objective of this study was to address the impact of heterogeneity of infectious period and contagiousness on Salmonella transmission dynamics in dairy cattle populations. We developed three deterministic SIR-type models with two basic infected stages (clinically and subclinically infected). In addition, model 2 included long-term shedders, which were defined as individuals with low contagiousness but long infectious period, and model 3 included super-shedders (individuals with high contagiousness and long infectious period). The simulated dynamics, basic reproduction number (R0) and critical vaccination threshold were studied. Clinically infected individuals were the main force of infection transmission for models 1 and 2. Long-term shedders had a small impact on the transmission of the infection and on the estimated vaccination thresholds. The presence of super-shedders increases R0 and decreases the effectiveness of population-wise strategies to reduce infection, making necessary the application of strategies that target this specific group.

Listeria monocytogenes fecal shedding in dairy cattle shows high levels of day-to-day variation and includes outbreaks and sporadic cases of shedding of specific L. monocytogenes subtypes.

Fecal shedding of Listeria monocytogenes poses a risk for contamination of animal feed and agricultural environments and raw food at the pre-harvest stages of food production. To be able to reduce these risks it is critical to improve understanding of the epidemiology of L. monocytogenes shedding in feces. The objective of this study was to assess the daily variability of fecal shedding and its association with individual animal (lactation number and the day of current lactation) and environmental (feed) risk factors. That was achieved by application of longitudinal daily sample collection in a herd of dairy cattle and molecular characterization of isolated L. monocytogenes. Fecal samples (25) and silage samples (2) were collected daily during two 2-week periods and one 5-day period. L. monocytogenes was isolated from 255 out of 825 (31%) fecal samples on 24 out of 33 (73%) days, and from 25 out of 66 (38%) silage samples on 16 out of 33 (48%) days. Ninety-four percent of cows excreted L. monocytogenes in feces at least once during the study period. Our data analyses indicated that (i) the prevalence and incidence risk of L. monocytogenes fecal shedding in cattle vary considerably over time, from 0 to 100%, and both are associated with contamination of silage, (ii) L. monocytogenes fecal shedding in cattle could occur as part of an outbreak or as an isolated sporadic case, (iii) L. monocytogenes subtypes associated with human infections are commonly isolated from cattle feces and silage, and (iv) a single cow can harbor more than one L. monocytogenes subtype on any given day. Although limited to a single dairy cattle herd, these findings provide a significant advancement in the understanding of the epidemiology of L. monocytogenes fecal shedding in dairy cattle.

The use of comparative duplex PCR in monitoring of patients with non-Hodgkin's lymphoma and chronic lymphocytic leukaemia.

Various quantitative PCR approaches have been utilized during the last years to provide information about the treatment efficacy and the risk of recurrent disease in haematological malignancies. Apart from the frequently used real-time PCR, cost-saving modified standard PCR methods may be applied as well. This report evaluates the utility of the end-point comparative duplex PCR. We have used this method for monitoring of 35 patients with either NHL or CLL and observed a good correlation between quantitative molecular results and clinical outcome. There was also an agreement between comparative duplex PCR and real-time PCR in patients who were monitored by both methods. We therefore believe that use of this technique should be strongly considered instead of simple qualitative detection in monitoring of therapeutic outcome in NHL or CLL patients.

[Clinical importance of semi-quantitative monitoring of lymphomas using the comparative polymerase chain reaction]. / Klinická relevance semikvantitativního monitorování lymfomu uzitím komparativní polymerázové retezové reakce.

BACKGROUND: PCR techniques detecting interchromosomal translocation and clonal immunoglobulin gene rearrangement (IgH) as disease markers in non-Hodgkin's lymphomas (NHL) has been utilised past ten years. However, qualitative PCR detection of persisted minimal residual disease cannot provide clinically useful prognostic information and presently, quantitative approaches are required to predict patient outcome and assess response to the treatment. In some cases, "end-point" quantifying techniques, such as comparative PCR, are applicable and the relative estimation of differences in target quantity may serve in disease monitoring rather than absolute number of target copies. METHODS AND RESULTS: Our method of comparative PCR employs co-amplification of sequences of interest (clonal CDR3, bcl2/Jh) and the segment of Hras 1 gene(ras) as an internal standard. Serial dilutions of stored diagnostic DNAs from blood and bone marrow are examined in the same PCR and, after gel densitometry, the amount of initial target is assessed by comparing exponential products of co-amplification. The comparative PCR assay was utilized in monitoring of NHL patients cured either with conventional therapy, or with high-dose regimens and transplantation with stem cells, or with chimaeric anti-CD20 monoclonal antibody (Rituximab). Results from 50 monitored intervals obtained during several months up to several years were supplemented with clinical statements retrospectively. Some of patients became PCR-negative, reappearance of PCR-positivity was observed as well. The decrease or increase of disease marker corresponded to clinical observations. Results obtained from bone marrow were in agreement with those obtained from blood. CONCLUSIONS: End-point quantifying PCR comparative assay may provide an information on the increased risk of relapse and impact of the therapy. The predictive value of these methods depends on the frequency of sample taking and on the sensitivity of the method, which should be monitored in negative cases.