Development of a cost-efficient micro-detection slide system for the detection of multiple shrimp pathogens.

In view of the current demand for rapid detection and identification of pathogens, point-of-care testing (POCT) with fast portability, low consumption, and increased sensitivity and specificity has become more and more popular. The emerging nucleic acid isothermal amplification technology (NAIAT) has shown potential advantages in the development of rapid microbial detection. In this study, a micro-detection slide system was developed based on the NAIAT of various nucleic acids of shrimp pathogens. The system included a micro-detection slide with 48 identical detecting cells precoated with all detection reagents, except the sample template. The process of producing the micro-detection slides mainly combined super-hydrophobic/super-oleophobic and super-hydrophilic materials to obtain separated spaces for detection, and aerosol pollution was eliminated in the form of water-in-oil. The micro-detection slide system was capable of simultaneously detecting 4 groups of samples and 8 important shrimp pathogens and is a relatively low-cost, portable, and high-throughput nucleic acid (RNA and DNA) detection technology. The establishment of this technology will provide key technical support for the construction of biosecurity systems for healthy shrimp culture.

Core genome multi-locus sequence typing as an essential tool in a high-cost livestock-associated meticillin-resistant Staphylococcus aureus CC398 hospital outbreak.

BACKGROUND: Livestock-associated meticillin-resistant Staphylococcus aureus (LA-MRSA) clonal complex (CC) 398 may be transmitted and cause morbidity and mortality in hospitals. The economic cost of stopping hospital transmission of LA-MRSA CC398 is poorly described. Early detection of transmission may limit the extent of the intervention. AIM: To evaluate core genome multi-locus sequence typing (cgMLST) for detecting transmission chains and to estimate the costs for interventions to prevent further spread after discovery of hospital transmission of LA-MRSA CC398. METHODS: Five patients were involved in two episodes of transmission of LA-MRSA CC398 in a hospital. Standard interventions including MRSA screening of patients and healthcare workers were initiated. Whole genome sequences of the five isolates and 17 epidemiologically unrelated MRSA CC398 isolates from other hospitalized patients were analysed by single nucleotide polymorphism (SNP) comparisons and cgMLST. The economic costs of constraining transmission were calculated from relevant sources. FINDINGS: The five isolates suspected to be involved in hospital transmission clustered with ≤2 SNPs in the draft genome sequences with some distance to other isolates. cgMLST allocated the five isolates to the same type, which was different from all but two of the sporadic isolates. Furthermore, cgMLST separated the five transmission isolates from all other isolates. The economic costs of the outbreak interventions exceeded €11,000 per patient. CONCLUSION: LA-MRSA CC398 is transmittable in hospitals, and intervention against transmission may reach considerable costs. cgMLST is useful in surveillance of hospital transmission of LA-MRSA.

Benefits and Costs of Animal Virulence for Microbes.

This essay is written from the vantage point of the microbial world. While the focus of much thought in the microbial pathogenesis and infectious diseases fields has been on the impact of host-microbe interaction on the host, here we ask questions about what happens to the microbe. What are the costs and benefits for microbes of having the capacity for virulence? Our exploration of this topic leads us to conclude that virulence confers very few benefits for microbes, unless disease is necessary for microbial survival through host-to-host spread. In fact, the capacity for virulence is often fraught with risk for microbes, including host dependence and the threat of extinction. The costs of virulence may explain why, relative to their enormous numbers in nature, very few microbes are actually associated with human and animal disease.

Assessment of a pan-dermatophyte nested-PCR compared with conventional methods for direct detection and identification of dermatophytosis agents in animals.

Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture-positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies.

Body size influences energetic and osmoregulatory costs in frogs infected with Batrachochytrium dendrobatidis.

Sloughing maintains the skins integrity and critical functionality in amphibians. Given the behavioural, morphological and osmoregulatory changes that accompany sloughing, this process is likely to be physiologically costly. Chytridiomycosis, a cutaneous disease of amphibians caused by the fungus Batrachochytrium dendrobatidis (Bd), disrupts skin function and increases sloughing rates. Moreover, mortality rates from chytridiomycosis are significantly higher in juveniles and so we hypothesised that smaller individuals maybe more susceptible to chytridiomycosis because of allometric scaling effects on the energetic and osmoregulatory costs of sloughing. We measured in-vivo cutaneous ion loss rates and whole animal metabolic rate (MR) of Green tree frogs, Litoria caerulea, over a range of body sizes both infected and uninfected frogs during sloughing. Infected animals had a greater rate of ion loss and mass-specific MR during non-sloughing periods but there were no additional effects of sloughing on either of these parameters. There were also significant interactions with body size and Bd load indicating that smaller animals with higher Bd loads have greater rates of ion loss and higher energetic demands. Our results shed light on why smaller Bd-infected anurans often exhibit greater physiological disruption than larger individuals.

Livestock-associated meticillin-resistant Staphylococcus aureus (MRSA) among human MRSA isolates, European Union/European Economic Area countries, 2013.

Currently, surveillance of livestock-associated meticillin-resistant Staphylococcus aureus (LA-MRSA) in humans in Europe is not systematic but mainly event-based. In September 2014, the European Centre for Disease Prevention and Control (ECDC) initiated a questionnaire to collect data on the number of LA-MRSA from human samples (one isolate per patient) from national/regional reference laboratories in European Union/European Economic Area (EU/EEA) countries in 2013. Identification of LA-MRSA as clonal complex (CC) 398 by multilocus sequence typing (MLST) was preferred, although surrogate methods such as spa-typing were also accepted. The questionnaire was returned by 28 laboratories in 27 EU/EEA countries. Overall, LA-MRSA represented 3.9% of 13,756 typed MRSA human isolates, but it represented ≥ 10% in five countries (Belgium, Denmark, Spain, the Netherlands and Slovenia). Seven of the reference laboratories did not type MRSA isolates in 2013. To monitor the dispersion of LA-MRSA and facilitate targeted control measures, we advocate periodic systematic surveys or integrated multi-sectorial surveillance.

In Vitro Assessment of the Probiotic Potential of Lactococcus lactis LMG 7930 against Ruminant Mastitis-Causing Pathogens.

Mastitis in dairy ruminants is considered to be the most expensive disease to farmers worldwide. Recently, the intramammary infusion of lactic acid bacteria has emerged as a potential new alternative to antibiotics for preventing and treating bovine mastitis. In this study we have investigated in vitro the probiotic potential of Lactococcus lactis LMG 7930, a food-grade and nisin-producing strain, against mastitis-causing pathogens. We have characterized its carbohydrate fermentation and antibiotic susceptibility profiles, cell surface properties and antimicrobial activity, as well as its capabilities to adhere to and inhibit the invasion of pathogens into the bovine mammary epithelial cell line BME-UV1d. We found that L. lactis LMG 7930 was sensitive to tested drugs, according to the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP), and showed an improved carbohydrate fermentation capacity compared to starter strains. Moreover, the strain exhibited antagonistic properties towards many of the pathogens tested. It presented medium surface hydrophobicity, a low basic property and no electron acceptor capability. It showed low auto-aggregation and no co-aggregation abilities towards any of the tested pathogens. The strain was one of the most adhesive to bovine mammary epithelial cells among tested bacteria, but its internalisation was low. The strain did not affect significantly pathogen invasion; however, a trend to decrease internalization of some pathogens tested was observed. In conclusion, our results suggest that this strain might be a promising candidate for the development of new strategies of mastitis control in ruminants. Future investigations are needed to evaluate its safety and efficacy under field conditions.

The evidentiary value of challenge trials for three pre-harvest food safety topics: a systematic assessment.

Reducing zoonotic pathogens in food animals prior to harvest will reduce the pathogen burden that enters the food chain and the environment. Consequently, the burden of enteric illness in humans may be reduced. Evaluating interventions to reduce a pathogen in animals often begins with challenge trials, in which animals are deliberately exposed to the pathogen under controlled conditions. Challenge trials are subsequently followed by field trials, also known as randomized controlled trials, in which the animals are naturally exposed to the pathogen. Challenge trials can most effectively inform field trials only if they precede field trials, are robust, internally valid and transparently reported. Using systematic review and meta-analysis methodology, we examined the pre-harvest food safety literature for three intervention-pathogen-species combinations: probiotics/competitive exclusion products in ruminants to reduce Escherichia coli O157 shedding, vaccines in ruminants to reduce E. coli O157 shedding and vaccines in swine to reduce Salmonella shedding. We examined two outcomes, prevalence of faecal shedding at the end of the trial and prevalence of faecal shedding throughout the trial period, to compare challenge trials and field trials. We found that challenge trials occurred concurrently with field trials, challenge trials suffered from reporting deficiencies of methodological features, challenge trials tended to report a more favourable outcome than field trials, and there was some evidence of publication bias among all three intervention-pathogen-species combinations. Challenge trials would better serve to inform field trials if they precede field trials, are methodologically sound, include transparent reporting and are published regardless of their results. In addition, due to our findings of greater efficacy reported among challenge trials compared with field trials, risk models predicting the public health benefits of pre-harvest interventions to reduce zoonotic pathogens in livestock might be best served by field trial results alone.

High-throughput sequencing in veterinary infection biology and diagnostics.

Sequencing methods have improved rapidly since the first versions of the Sanger techniques, facilitating the development of very powerful tools for detecting and identifying various pathogens, such as viruses, bacteria and other microbes. The ongoing development of high-throughput sequencing (HTS; also known as next-generation sequencing) technologies has resulted in a dramatic reduction in DNA sequencing costs, making the technology more accessible to the average laboratory. In this White Paper of the World Organisation for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine (Uppsala, Sweden), several approaches and examples of HTS are summarised, and their diagnostic applicability is briefly discussed. Selected future aspects of HTS are outlined, including the need for bioinformatic resources, with a focus on improving the diagnosis and control of infectious diseases in veterinary medicine.

A historical review of managed honey bee populations in Europe and the United States and the factors that may affect them.

Honey bees are a highly valued resource around the world. They are prized for their honey and wax production and depended upon for pollination of many important crops. While globally honey bee populations have been increasing, the rate of increase is not keeping pace with demand. Further, honey bee populations have not been increasing in all parts of the world, and have declined in many nations in Europe and in North America. Managed honey bee populations are influenced by many factors including diseases, parasites, pesticides, the environment, and socio-economic factors. These factors can act alone or in combination with each other. This review highlights the present day value of honey bees, followed by a detailed description of some of the historical and present day factors that influence honey bee populations, with particular emphasis on colony populations in Europe and the United States.

Engineering control of airborne disease transmission in animal laboratories.

We here present a review of the problem of controlling airborne disease transmission in animal research facilities, with emphasis on engineering design and air-treatment technologies. Dilution ventilation, pressurization control, source control, and air disinfection and removal systems are reviewed, and analytical studies on the effects of dilution ventilation, filtration, and ultraviolet germicidal irradiation are summarized. In addition, we discuss practical problems common to laboratory facilities and present a database of potential airborne pathogens and allergens that can be transmitted between humans and animals. We offer some conclusions regarding the design and selection of available technologies and components and provide cost estimates for various air-cleaning systems.

DANMAP: monitoring antimicrobial resistance in Denmark.

The objectives of the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) are to monitor trends in resistance among bacteria from animals, food and humans, to monitor the consumption of antimicrobial agents and to determine the association between consumption and occurrence of resistance and to model transmission of resistance from animals to humans. DANMAP is based on the examination of representative bacterial isolates of animal and human pathogens, of zoonotic bacteria and of indicator bacteria. For food animals, both diseased and the healthy populations are studied. Isolates from all three reservoirs are examined for their susceptibility to a basic panel of antibiotics that includes representatives of the major classes of compounds, making comparison of resistance levels in the reservoirs possible. Isolates are stored in a strain collection and are available for further study. The data are stored in databases as MIC values or mm inhibition zones with all identifiers. A system for recording all use of the veterinary medicines, VETSTAT, is currently under implementation. For production animals, the consumption will be recorded for each herd, providing a basis for detailed modelling of the effect of consumption on resistance.

Antibiotic resistance in Spain: what can be done? Task Force of the General Direction for Health Planning of the Spanish Ministry of Health.

The document Antibiotic Resistance in Spain: What Can Be Done? was prepared in 1994 by an ad hoc Task Force under the auspices of the Spanish Ministry of Health. The problem of the development of antibiotic resistance in community-acquired bacterial pathogens in Spain and its relation to antibiotic use and consumption has been of both national and international concern. This document analyzes the problem and provides general recommendations for limiting the emergence, evolution, and spread of resistant microorganisms. Particular emphasis is given to systems for surveillance of antibiotic resistance, the monitoring of patterns of antibiotic use and consumption, and strategies for influencing the producers (pharmaceutical industry), prescribers (physicians and veterinarians), dispensers (pharmacists), and consumers (patients) of antibiotics. Control of the emergence of antibiotic resistance requires a concerted effort on the part of all social and scientific agencies involved in health care.