Growth potential of faecal bacteria in simulated psychrophilic/mesophilic zones during composting of organic waste

Aim: This study investigated the growth potential of Salmonella serotype Typhimurium and faecal indicator organisms in compost materials and the correlation between bacterial growth potential and the physico‐chemical composition of the compost substrate and temperature. Methods and Results: Survival of Salm. Typhimurium, Enterococcus spp. and total coliforms at 14, 24 and 37°C was determined in material of different degrees of maturity collected from composting plants for household waste and manure. All three micro‐organisms showed the potential for growth in the material from active composts (Solvita index 4) but inactivation generally occurred over time in mature compost material (Solvita index 7–8). Conclusions: Salm. Typhimurium had the potential for growth in psychrophilic/mesophilic (P/M) zones of immature compost material and its growth potential correlated negatively with the maturity of the compost and the temperature within the simulated P/M zone. Significance and Impact of the Study: The risk of pathogen regrowth in P/M zones during organic waste composting further emphasizes the importance of good management practices and of avoiding P/M zones in combination with low compost maturity.     

Treatment with Ca(OH)2 for inactivation of Salmonella Typhimurium and Enterococcus faecalis in soil contaminated with infected horse manure

Aim: To investigate the inactivation of Salmonella enterica serovar Typhimurium and the faecal indicator Enterococcus faecalis in horse manure:soil mixtures by application of hydrated lime (Ca(OH)₂). Methods and Results: In laboratory incubations, the inhibitory effect of different concentrations of Ca(OH)₂, as well as different application techniques, was tested. Other variables were horse manure:soil ratio, incubation temperature (6 and 14°C) and soil type (sand/clay). Bacterial enumeration by the plate count method in samples taken at increasing intervals revealed that Ca(OH)₂ effectively reduced Salmonella Typhimurium numbers. However, to achieve a sufficient reduction, the Ca(OH)₂ had to be applied at a sufficient rate, and the amount required varied because of manure:soil ratio and incubation temperature. The results showed that a pH above 11 was needed and that a high pH had to be maintained for up to 7 days. An appropriate application technique for the Ca(OH)₂ was also important, so that a high pH was obtained throughout the whole material to be treated. In addition, a high manure:soil ratio in combination with a higher incubation temperature was found to rapidly neutralize the pH and to increase the risk of Salmonella re‐growth. Conclusions: Application of Ca(OH)₂ can be an efficient method for treating a Salmonella‐contaminated horse paddock. A high pH is a key factor in Salmonella inactivation, and thus, monitoring the pH during the treatment period is necessary. To avoid re‐growth excess manure should be removed for separate treatment elsewhere. Significance and Impact of the Study: Persistence of Salmonella in horse paddocks poses a risk of disease transmission to healthy animals and people who come into contact with these animals. An efficient method to de‐contaminate a Salmonella‐contaminated soil would be a valuable tool for animal welfare and for public health.     

Target-Dependent Enrichment of Virions Determines the Reduction of High-Throughput Sequencing in Virus Discovery

Viral infections cause many different diseases stemming both from well-characterized viral pathogens but also from emerging viruses, and the search for novel viruses continues to be of great importance. High-throughput sequencing is an important technology for this purpose. However, viral nucleic acids often constitute a minute proportion of the total genetic material in a sample from infected tissue. Techniques to enrich viral targets in high-throughput sequencing have been reported, but the sensitivity of such methods is not well established. This study compares different library preparation techniques targeting both DNA and RNA with and without virion enrichment. By optimizing the selection of intact virus particles, both by physical and enzymatic approaches, we assessed the effectiveness of the specific enrichment of viral sequences as compared to non-enriched sample preparations by selectively looking for and counting read sequences obtained from shotgun sequencing. Using shotgun sequencing of total DNA or RNA, viral targets were detected at concentrations corresponding to the predicted level, providing a foundation for estimating the effectiveness of virion enrichment. Virion enrichment typically produced a 1000-fold increase in the proportion of DNA virus sequences. For RNA virions the gain was less pronounced with a maximum 13-fold increase. This enrichment varied between the different sample concentrations, with no clear trend. Despite that less sequencing was required to identify target sequences, it was not evident from our data that a lower detection level was achieved by virion enrichment compared to shotgun sequencing.     

Salmonella reduction in manure by the addition of urea and ammonia

Animal manure may contain pathogenic microorganisms and disinfection is suggested to avoid reinfection of animal herds and introduction of zoonotic disease into the food chain. Ammonia and urea were tested for disinfection of bovine manure and Salmonella was found to be rapidly eliminated by the addition of 0.5% aqueous ammonia or 2% w/w urea (s). Treatments (2% urea and 0.5% ammonia), temperature (4 degrees C or 14 degrees C) and combinations of these factors significantly affected the inactivation rate. Decimal values (T(90)) were reduced from 8.3 days in the control to 2.0 days and 0.4 days at 14 degrees C after the addition of urea and ammonia, respectively. At 4 degrees C, the decimal values were reduced from 34 to 4.8 and 1.1 days, respectively. Recommended treatments of bovine manure based on Monte Carlo simulations are 0.5% ammonia followed by storage for one week or 2% urea followed by storage for two weeks at 14 degrees C, one month at 4 degrees C. Storage without additives should include at least one summer in temperate regions. Enterococci were evaluated as indicators for Salmonella but significantly slower decay rate and different behaviour in the material made them unsuitable as indicators for Salmonella in manure disinfected by ammonia or urea. Free ammonia treatment of Salmonella-contaminated manure disinfects the material and raises its fertilizer value.     

Composting for avian influenza virus elimination

Effective sanitization is important in viral epizootic outbreaks to avoid further spread of the pathogen. This study examined thermal inactivation as a sanitizing treatment for manure inoculated with highly pathogenic avian influenza virus H7N1 and bacteriophages MS2 and 6. Rapid inactivation of highly pathogenic avian influenza virus H7N1 was achieved at both mesophilic (35°C) and thermophilic (45 and 55°C) temperatures. Similar inactivation rates were observed for bacteriophage 6, while bacteriophage MS2 proved too thermoresistant to be considered a valuable indicator organism for avian influenza virus during thermal treatments. Guidelines for treatment of litter in the event of emergency composting can be formulated based on the inactivation rates obtained in the study.