Spore Contamination Research Articles

Analysis of spore prevalence and sporulation potential in prepackaged meat products

Spores are recognized as significant factors affecting the safety of prepackaged meat products. However, current knowledge about the prevalence and distribution of spores in these products is limited. This study investigated 360 commonly consumed prepackaged meat products sourced from Zhengzhou, Henan Province, to assess spore prevalence, species, and characteristics. Using the 16 S rRNA method, primary spore species were isolated and identified. The results revealed a spore prevalence of 60.6% (218/360), identifying eight distinct bacterial species. In laboratory cultivation, Bacillus. Subtilis exhibited the highest spore sporulation efficiency at 95.1%, indicating a significant foodborne risk. Under the storage condition of −4 °C, the spore in food (starch sausage) maintained stable dormant state within 60 days. Additionally, Raman spectroscopy was employed to analyze the biochemical components of the cultured spores, accurately classifing eight species of spores using linear discriminant analysis (LDA). The high percentage of spore contamination in prepackaged meat products poses a potential health risk to consumers. An enhanced understanding of spore prevalence, sporulation potential, and storage methods in these products can empower industries to adjust measures effectively, controlling spore pollution in meat products.

Trophic facilitation in forest restoration: Can Nothofagus trees use ectomycorrhizal fungi of the pioneer shrub Leptospermum?

The benefits of plant-to-plant facilitation in ecological restoration are well recognized, yet the potential for indirect trophic facilitation remains understudied. Nothofagus (southern beech; Nothofagaceae) is an iconic southern hemisphere tree genus that is frequently the focus of ecological restoration efforts. One aspect of Nothofagus ecology that may limit restoration success is the availability of appropriate ectomycorrhizal fungi. It has been suggested that pioneer dual-mycorrhizal hosts such as Leptospermum species (Myrtaceae) could facilitate Nothofagus establishment by providing fungal inoculum, but the capacity for Nothofagus to use Leptospermum ectomycorrhizal fungi is unknown. To investigate potential indirect facilitation, we conducted a common garden pot trial to determine if Nothofagus cliffortioides (mountain beech) can use symbionts from Leptospermum scoparium (mānuka) ectomycorrhizal communities. Nothofagus and Leptospermum seedlings were grown in monoculture and mixed pairs with reciprocal "home" and "away" soil fungal inoculum. ITS2 metabarcoding of eDNA from hyphal ingrowth bags revealed that Nothofagus and Leptospermum inoculum contained different ectomycorrhizal fungal communities, but that half of the common ectomycorrhizal taxa identified were found in both soil types, suggesting generalist fungi exist. Nothofagus was able to form associations with some fungal species originating from Leptospermum inoculum, however, probable spore contamination meant that the proportion of root colonization associated with those species was ambiguous. Root ectomycorrhizal colonization rates were positively associated with seedling biomass, and there was some evidence of a home soil inoculum advantage in Nothofagus, but these effects were minor. Additionally, we found evidence that home inoculum provides a protective advantage against drought stress for Leptospermum seedlings. Our results indicate the potential for using Leptospermum to promote Nothofagus establishment in restoration plantings and highlight the possible benefits of considering fungal mutualists in ecological restoration projects.

Inactivation Effect of Germination Combined with Cold Plasma Treatment on Bacillus licheniformis Spores.

Food spoilage, primarily caused by spore-forming bacteria, has become a critical concern since it results in substantial economic losses within the food industry. Past investigations have successfully identified Bacillus licheniformis as the main bacterium responsible for spoilage in roast chicken. In this study, we screened a new sterilization combination from 16 germinants and 4 cold plasma conditions, respectively. Among them, the combination of "A"GFNa-1 (composed of 60 mmol/L L-alanine, 10 mmol/L D-glucose, 10 mmol/L D-fructose, and 1 g/L NaCl) with cold plasma treatment (packed with 100% argon at 70 kV) proved effective in deactivating B. licheniformis spores, resulting in a reduction of approximately 2.1 log CFU/mL. Furthermore, we exposed the spores to different conditions: CK (no germination, no cold plasma), MF (germination only), CP (no germination, 100% argon packed, 70 kV cold plasma treatment for 3 min), and MF + CP (germination for 5 h, 100% argon packed, 70 kV cold plasma treatment for 3 min). The results of heat inactivation and dipicolinic acid (DPA) release rate demonstrated that cold plasma treatment effectively inactivated both spores and vegetative cells without inducing germination. Additionally, the reduced survival under hyperosmotic conditions and the presence of distinct red fluorescence patterns observed through confocal laser scanning microscopy (CLSM) collectively suggest that cold plasma treatment disrupts the inner membrane structure and leads to the inactivation of B. licheniformis. Overall, our findings indicate a spore clearance rate of 99.2% and suggest that the combination of efficient germinants and cold plasma treatment holds promise as a viable approach to mitigate spore contamination in the food industry.

Quantitative microbial spoilage risk assessment caused by fungi in sports drinks through multilevel modelling

The risk of fungal spoilage of sports drinks produced in the beverage industry was assessed using quantitative microbial spoilage risk assessment (QMSRA). The most relevant pathway was the contamination of the bottles during packaging by mould spores in the air. Mould spores' concentration was estimated by longitudinal sampling for 6 years (936 samples) in different production areas and seasons. This data was analysed using a multilevel model that separates the natural variability in spore concentration (as a function of sampling year, season, and area) and the uncertainty of the sampling method. Then, the expected fungal contamination per bottle was estimated by Monte Carlo simulation, considering their settling velocity and the time and exposure area. The product's shelf life was estimated through the inoculation of bottles with mould spores, following the determination of the probability of visual spoilage as a function of storage time at 20 and 30 °C using logistic regression. The Monte Carlo model estimated low expected spore contamination in the product (1.7 × 10−6 CFU/bottle). Nonetheless, the risk of spoilage is still relevant due to the large production volume and because, as observed experimentally, even a single spore has a high spoilage potential. The applicability of the QMSRA during daily production was made possible through the simplification of the model under the hypothesis that no bottle will be contaminated by more than one spore. This simplification allows the calculation of a two-dimensional performance objective that combines the spore concentration in the air and the exposure time, defining “acceptable combinations” according to an acceptable level of spoilage (ALOS; the proportion of spoiled bottles). The implementation of the model at the operational level was done through the representation of the simplified model as a two-dimensional diagram that defines acceptable and unacceptable areas. The innovative methodology employed here for defining and simplifying QMSRA models can be a blueprint for future studies aiming to quantify the risk of spoilage of other beverages with a similar scope.

Zirconium-Directed Supramolecular Self-Assembly of Coenzyme A@GNCs with Enhanced Phosphorescence for Developing Ultrasensitive Tracer Probe of Dipicolinic Acid, a Biomarker of Bacterial Spores.

Luminescent gold nanoclusters (GNCs) are a class of attractive quantum-sized nanomaterials bridging the gap between organogold complexes and gold nanocrystals. They typically have a core-shell structure consisting of a Au(I)-organoligand shell-encapsulated few-atom Au(0) core. Their luminescent properties are greatly affected by their Au(I)-organoligand shell, which also supports the aggregation-induced emission (AIE) effect. However, so far, the luminescent Au nanoclusters encapsulated with the organoligands containing phosphoryl moiety have rarely been reported, not to mention their AIE. In this study, coenzyme A (CoA), an adenosine diphosphate (ADP) analogue that is composed of a bulky 5-phosphoribonucleotide adenosine moiety connected to a long branch of vitamin B5 (pantetheine) via a diphosphate ester linkage and ubiquitous in all living organisms, has been used to synthesize phosphorescent GNCs for the first time. Interestingly, the synthesized phosphorescent CoA@GNCs could be further induced to generate AIE via the PO32- and Zr4+ interactions, and the observed AIE was found to be highly specific to Zr4+ ions. In addition, the enhanced phosphorescent emission could be quickly turned down by dipicolinic acid (DPA), a universal and specific component and also a biomarker of bacterial spores. Therefore, a Zr4+-CoA@GNCs-based DPA biosensor for quick, facile, and highly sensitive detection of possible spore contamination has been developed, showing a linear concentration range from 0.5 to 20 μM with a limit of detection of 10 nM. This study has demonstrated a promising future for various organic molecules containing phosphoryl moiety for the preparation of AIE-active metal nanoclusters.

Identification of Clinical Isolates of <i>Bacillus cereus</i> Group and Their Characterization by Mass Spectrometry and Electron Microscopy

Bacillus cereus is a spore-forming bacterium found in the environment mainly in soil. Bacillus spores are known to be extremely resistant not only to environmental factors, but also to various sanitation regimes. This leads to spore contamination of toxin-producing strains in hospital and food equipment and, therefore, poses a great threat to human health. Two clinical isolates identified as B. cereus and B. cytotoxicus were investigated in the present work. It was shown that their calcium ion content was significantly lower than that of the reference strains. According to electron microscopy, one of the isolates SRCC 19/16 has an enlarged exosporium, and isolate SRCC 1208 shows large electron-dense inclusions of unclear nature during sporulation. We can assume that it contains a biologically active component with a cytotoxic effect and possibly plays a role in pathogenesis. Comparative chemical, biochemical, physiological, and ultrastructural analysis of spores of clinical isolates and reference strains of B. cereus was performed. The results obtained deepen our understanding of the properties of spores that contribute to the increased pathogenicity of B. cereus group species.

Use of Dicranum polysetum extract against Paenibacillus larvae causing American Foulbrood under in vivo and in vitro conditions.

Recent research shows that Dicranum species can be used to ameliorate the negative effects of honeybee bacterial diseases and that novel compounds isolated from these species may have the potential to treat bacterial diseases. This study aimed to investigate the efficacy of Dicranum polysetum Sw. against American Foulbrood using toxicity and larval model. The effectiveness of D. polysetum Sw. ethanol extract in combating AFB was investigated in vitro and in vivo. This study is important in finding an alternative treatment or prophylactic method to prevent American Foulbrood disease in honey bee colonies. Spore and vegetative forms of Paenibacillus larvae PB31B with ethanol extract of D. polysetum were tested on 2040 honey bee larvae under controlled conditions. Total phenolic and flavonoid contents of D. polysetum ethanol extracts were determined as 80.72mg/GAE(Gallic acid equivalent) and 303.20µg/mL, respectively. DPPH(2,2-diphenyl-1-picrylhydrazyl) radical scavenging percent inhibition value was calculated as 4.32%. In Spodoptera frugiperda (Sf9) and Lymantria dispar (LD652) cell lines, the cytotoxic activities of D. polysetum extract were below 20% at 50µg/mL. The extract was shown to considerably decrease infection in the larvae, and the infection was clinically halted when the extract was administered during the first 24h after spore contamination. The fact that the extract contains potent antimicrobial/antioxidant activity does not reduce larval viability and live weight, and does not interact with royal jelly is a promising development, particularly regarding its use to treat early-stage AFB infection.

Feral Swine as Indirect Indicators of Environmental Anthrax Contamination and Potential Mechanical Vectors of Infectious Spores.

Anthrax is a disease that affects livestock, wildlife, and humans worldwide; however, its relative impacts on these populations remain underappreciated. Feral swine (Sus scrofa) are relatively resistant to developing anthrax, and past serosurveys have alluded to their utility as sentinels, yet empirical data to support this are lacking. Moreover, whether feral swine may assist in the dissemination of infectious spores is unknown. To address these knowledge gaps, we intranasally inoculated 15 feral swine with varying quantities of Bacillus anthracis Sterne 34F2 spores and measured the seroconversion and bacterial shedding over time. The animals also were inoculated either one or three times. The sera were evaluated by enzyme-linked immunosorbent assay (ELISA) for antibodies against B. anthracis, and nasal swabs were cultured to detect bacterial shedding from the nasal passages. We report that the feral swine developed antibody responses to B. anthracis and that the strength of the response correlated with the inoculum dose and the number of exposure events experienced. Isolation of viable bacteria from the nasal passages of the animals throughout the study period suggests that feral swine may assist in the spread of infectious spores on the landscape and have implications for the identification of environments contaminated with B. anthracis as well as the exposure risk to more susceptible hosts.

Ultrastructural observation and transcriptome analysis provide insights into mechanisms of Penicillium expansum invading apple wounds

Penicillium expansum is a pathogen causing enormous postharvest losses of fruits, especially apples. In this study, we first investigated the morphological changes of P. expansum within apple wounds during infectious process by microscopic observation. We found that conidia swelled and secreted potential hydrophobin in 4 h, germinated in 8 h, and finally formed conidiophores in 36 h, a critical control time point to prevent the second contamination of spores. We then compared the transcript accumulation of P. expansum in apple tissues and liquid culture at 12 h. In total, 3168 and 1318 up-regulated and down-regulated genes were identified. Among them, genes regarding the biosynthesis of substances such as ergosterol, organic acid, cell wall degrading enzymes, and patulin were induced in expression. Pathways were activated, including autophagy, the mitogen-activated protein kinase, and pectin degradation. Our findings provide insights into the lifestyle and the mechanisms of P. expansum invading apple fruits.

Identification of Clinical Isolates of the Bacillus cereus Group and Their Characterization by Mass Spectrometry and Electron Microscopy

Bacillus cereus is a spore-forming bacterium found in the environment mainly in soil. Bacillus spores are known to be extremely resistant not only to environmental factors, but also to various sanitation regimes. This leads to spore contamination of toxin-producing strains in hospital and food equipment and, therefore, poses a great threat to human health. Two clinical isolates identified as B. cereus and B. cytotoxicus were used in the present work. It was shown that their calcium ion content was significantly lower than that of the reference strains. According to electron microscopy, one of the SRCC 19/16 isolates has an enlarged exosporium, and the SRCC 1208 isolate has large electron-dense inclusions of an unclear nature during sporulation. We can assume that these contain a biologically active component with a cytotoxic effect and possibly play a role in pathogenesis. Comparative chemical, biochemical, physiological, and ultrastructural analysis of spores of clinical isolates and reference strains of B. cereus was performed. The results we obtained deepen our understanding of the properties of spores that contribute to the increased pathogenicity of B. cereus group species.

What’s New in Prevention of Invasive Fungal Diseases during Hospital Construction and Renovation Work: An Overview

The goal of the overview was to give insight into the recent data of invasive fungal diseases (IFDs) associated with construction and renovation in healthcare settings as well as the recent evidence about available prevention and infection control measures. The number of studies describing IFD outbreaks associated with construction or renovation is on the rise again. Applying adequate prevention measures is still a challenge not just for healthcare workers but also for architects and construction workers as well. The role of multidisciplinary teams in the planning and monitoring of prevention measures cannot be overemphasized. Dust control is an inevitable part of every prevention plan. HEPA filters are helpful in the prevention of fungal outbreaks in hematologic patients, but further studies are needed to clarify the extent in which they contribute as specific control measures. The cut-off value for a "threating" level of fungal spore contamination still remains to be defined. The value of antifungal prophylaxis is difficult to assess because other preventive measures are simultaneously applied. Recommendations are still based on few meta-analyses, a large number of descriptive reports, and the opinion of respective authorities. Outbreak reports in the literature are a valuable resource and should be used for education as well as for preparing outbreak investigations.

Microbiological, Physicochemical and Sensory Characterization of Honey, a Natural Healthy Product in Burkina Faso

Honey is a natural product produced by bees from the nectar of flowers. It is a very healthy food whose multiple properties significantly improve health and prevent many diseases. However, some practices can affect its quality, hence the objective of the study was to assess the honey safety from 6 honey-producing regions in Burkina Faso. The physicochemical, microbiological and sensory characteristics were determined using standard methods.
 The density was between 1.39 to 1.44; the pH, 5.73 to 6.56; the total acidity, 13.00 to 83.00 meq/kg; the brix degree ranged 86.00 to 88.50%; the moisture,11.86 to 18.83%, the electrical conductivity, 101.00 to 155.00 μs/cm and the Hydroxymethylfurfural (HMF), from 14.67± 0.00 to 90.52±0.35.
 Microbiological analysis showed the total counts varied from 1.21.105±1.18.104 to 3.50.103±3.50.103 to 1.21.105 CFU/mL; yeast and mold rate were below 103 CFU/mL, spore contamination is also noted in some honey samples and was between 2.23×101±2.51 to 1.38×102 ±7.63CFU/mL), Salmonella, Shigella and coliform were not detected in the honey samples.
 Sensory analysis revealed that the organoleptic characteristics of honey varied from region to another. All the honey was differently appreciated by the tasters.

Dose-response analysis of Bacillus thuringiensis HD-1 cry- spore reduction on surfaces using formaldehyde with pre-germination.

To establish a basis for rapid remediation of large areas contaminated with Bacillus anthracis spores. Representative surfaces of wood, steel and cement were coated by nebulization with B. thuringiensis HD-1 cry- (a simulant for B. anthracis) at 5.9± 0.2, 6.3± 0.2 and 5.8± 0.2 log10 CFU per cm2 , respectively. These were sprayed with formaldehyde, either with or without pre-germination. Low volume (equivalent to ≤2500 L ha-1 ) applications of formaldehyde at 30 g l-1 to steel or cement surfaces resulted in ≥4 or ≤2 log10 CFU per cm2 reductions respectively, after 2h exposure. Pre-germinating spores (500 mmol l-1 l-alanine and 25 mmol l-1 inosine, pH7) followed by formaldehyde application showed higher levels of spore inactivation than formaldehyde alone with gains of up to 3.4 log10 CFU per cm2 for a given dose. No loss in B. thuringiensis cry- viability was measured after the 2h germination period, however, a pre-heat shock log10 reduction was seen for B. anthracis strains: LSU149 (1.7 log10), Vollum and LSU465 (both 0.9 log10), LSU442 (0.2 log10), Sterne (0.8 log10) and Ames (0.6 log10). A methodology was developed to produce representative spore contamination of surfaces along with a laboratory-based technique to measure the efficacy of decontamination. Dose-response analysis was used to optimize decontamination. Pre-germinating spores was found to increase effectiveness of decontamination but requires careful consideration of total volume used (germinant and decontaminant) by surface type. To be practically achievable, decontamination of a wide area contaminated with B. anthracis spores must be effective, timely and minimize the amount of materials required. This study uses systematic dose-response methodology to demonstrate that such an approach is feasible.

Selection of spore-specific aptamers for Geobacillus stearothermophilus, a food spoilage bacterium

Due to their ability to form extremely heat resistant spores, anaerobic bacteria are responsible for frequent food spoilage. The development of rapid and specific methods for the detection and quantification of spore contamination is therefore of major interest. In this paper, we describe for the first time the selection of aptamers specific to spores of Geobacillus stearothermophilus (Gbs), which induce flat sour spoilage in vegetable cans. Eighteen Spore-SELEX cycles were performed including 4 counter-selections with 12 bacteria commonly found in cannery. To optimise candidate amplification, PCR in emulsion was performed, and high-throughput sequencing analysis was applied to follow candidate evolution. Sequencing of aptamers from cycle 18 revealed 43 overrepresented sequences whose copy number exceeds 0.15% of the total obtained sequences. Within this group, the A01 aptamer presented a much higher enrichment with a relative abundance of 17.71%. Affinity and specificity for Gbs spores of the 10 most abundant candidates at cycle 18 were confirmed by PCR assay based on aptamer-spore complex formation and filtration step. Obtaining these aptamers is the starting point for the future development of biosensors dedicated to the detection of Gbs spores.

The sporicidal activity of a disinfectant with peracetic acid against the spores of Bacillus subtilis and Bacillus cereus according to the european standard PN-EN 17126: 2019-01.

The assessment of the sporicidal effectiveness of disinfectants is important from the point of view of the prevention of nosocomial infections and spore contamination of clinical samples, medical equipment and materials used in patient care. The rods of Bacillus spp. cause infections of the digestive system, bloodstream and, less often, respiratory tract. Cases were diagnosed in immunocompromised patients, malignant neoplasms and in neonatal wards. The source of the infection was hospital linen, reusable towels, catheters or milk from the human milk bank. Determination of the minimal sporicidal parameters of a disinfectant containing peracetic acid. The sporicidal activity of a disinfectant containing peracetic acid against Bacillus subtilis and Bacillus cereus spore suspensions was tested in a defined concentration range during a contact time of 15 minutes, in the presence of various interfering substances (clean and dirty conditions) according to the European Standard PN-EN 17126: 2019-01. The disinfecting preparation containing peracetic acid showed sporicidal activity against Bacillus subtilis at a concentration of 1% for 15 minutes under clean and dirty conditions and at concentrations of 0.5%, 1.00% and 1.25% against the Bacillus cereus spores during the same contact time but only under dirty conditions. The preparation showed no sporicidal activity against Bacillus cereus at concentrations of 2%, 3%, 4% and 5% during a contact time of 15 minutes under both dirty and clean conditions. In areas where there is a risk of infecting a patient or contaminating clinical specimens, materials and equipment with spores of Bacillus spp., it is necessary to use disinfectants with sporicidal activity confirmed according to the PN-EN 17126: 2019-01 standard. The sporicidal activity of disinfectants containing peracetic acid may depend on the method of preparing the solutions, their concentration, pH, temperature and the contamination degree of the disinfected surface.

Irrigation of Olives with Reclaimed Wastewaters and Deficit Strategies Affect Pathogenic Bacteria Contamination of Water and Soil.

This study aimed to evaluate pathogenic bacterial contamination of the water-soil-plant system in potted olive trees irrigated with reclaimed wastewater. Desalinated water (DW) obtained by treating municipal wastewater (SW) and reclaimed water (RW) obtained by mixing SW with the brine (BR) produced by DESERT technology (tertiary treatment by ultrafiltration, active carbon and reverse osmosis) were used. Two different irrigation regimes were compared: full irrigation (FI) and regulated deficit irrigation (RDI). During two irrigation seasons the concentrations of Escherichia coli, enterococci, spores of sulfite-reducing Clostridia (SRC) and Salmonella spp. were monitored in water, soil and fruit samples. Microbial concentrations in DW were always below the threshold for reuse in agriculture, while RW showed the highest level of contamination for all observed parameters. RDI management appeared to increase the soil content of SRC spores with respect to FI. Sporadically low SRC spore contamination was recorded in some fruits only in 2018, regardless of the irrigation source, probably because of accidental contamination during sampling or post-harvest handling. This study encourages the creation of a better regulatory framework reference, with specific guidelines for the use of RW as part of integrated environmental systems for the management of sustainable agriculture.

Multi-country surveillance of Clostridioides difficile demonstrates high prevalence of spores in non-healthcare environmental settings

BackgroundC. difficile spores are frequently isolated from hospital and non-healthcare settings but a worldwide analysis has not been done. The study objectives were to assess C. difficile spore contamination in the hospital and non-healthcare environments across a variety of countries. MethodsField studies assessed hospital vs. non-healthcare C. difficile spore contamination in hospitals, non-healthcare buildings, outdoor environments, and shoes. Swabs were cultured anaerobically for C. difficile and typed using PCR-fluorescent ribotyping. C. difficile contamination by swabbing area and geographic locations were compared. FindingsA total of 7,857 unique samples were collected primarily from the USA (89%) in addition to 9 other countries. The global prevalence of C difficile from environmental samples was 25.3% and did not differ between countries. In USA based studies, C. difficile contamination rates were similar for healthcare buildings (23.2%), non-healthcare buildings (23.4%), and outdoor spaces (24.7%). Floor samples had significantly higher (p < 0.001) C. difficile contamination rate (46.5%) followed by non-floor samples (21.1%), and bathrooms (15.3%). In a comparison of USA to other country samples, C. difficile contamination rates were similar for USA samples (21.5%) compared to rest of world samples (22.3%; p = 0.61). The most common ribotypes included F014-020 (15.7%), F106 (12.6%), F010 (8.9%), F027 (8.8%), and F002 (8.1%) and did not differ significantly between USA and non-USA samples. Finally, 546 of 1,218 (44.8%) shoe soles swabbed from the USA were contaminated with C. difficile spores. InterpretationThis large surveillance study of several countries demonstrated high prevalence of toxigenic C. difficile in non-healthcare environments with high contamination rates from floors and shoe soles.

Conditions Influencing Mould Growth for Effective Prevention of Wood Deterioration Indoors

Effective prevention of mould growth indoors is still an important topic considering that mould growth is frequently observed in buildings, it causes serious health hazards and can irreversibly damage infected objects. Several studies have been conducted and mould growth models developed. Despite that, some potentially important aspects such as water damage and spore contamination have received only little attention. The objective of the present study was to investigate the effect of the initial moisture content of wood and spore contamination on mould development indoors. The mould tests were performed in constant temperature (10, 20 and 30 °C) and relative humidity (91% and 97%) conditions. The results show that wetting of wood specimens prior to the test significantly accelerates mould growth at a temperature of 10 °C. For the other temperatures, the effect was insignificant. Similar results were obtained for the test involving dry (conditioned at RH 50%) and conditioned specimens (RH 91% or RH 97%). The results regarding initial spore contamination show that significantly longer periods are required for mould to develop without spore contamination at 10 °C and 20 °C, while at 30 °C the effect is relatively small.

Farm Silage Facilities and Their Management for the Prevention of Anaerobic Bacteria Spore Contamination in Raw Milk

At feed-out, aerobic spoilage of silage enables an increase in anaerobic spore-forming bacteria (ANSB) that may enter the total mixed ration (TMR). The aim of our study was to understand whether in hot summers the silage structures and management may affect the level of ANSB in milk for long-ripening cheese production. A survey of silage facilities, management, and their relationships with silage, TMR, feces, and milk ANSB most probable number (MPN) content was conducted in the Po Valley during summer months. Silo type did not affect the mean ANSB, but only the wideness of their value distributions, with a narrow range for bags and a wider range for bunkers. The unloading equipment affected the ANSB count; the front-end loader with cutter was associated with a lower ANSB count—probably as a result of the reduced surface left after daily silage removal. Silo length and daily removed face width were the main factors affecting contamination of silage by spore-forming bacteria during summer, with longer silos and wider surface removal reducing ANSB contamination—probably as a consequence of reduced aerobic spoilage at the silage surface. The silage contamination by spore-forming bacteria within a log10 2 MPN g−1 allowed a low concentration of spore-forming bacteria at the farm bulk milk tank level. Fecal ANSB levels did not factor into the regression that explains the ANSB in farm milk. It has been found that silage facilities’ features and their management are an important first step to reduce the extent of ANSB contamination at the farm level.

Data-based prediction of microbial contamination in herbs and identification of optimal harvest parameters

Abstract The quality of freshly harvested herbs is affected by several crucial factors, such as weather, tillage, fertilization, drying, and the harvesting process, e.g. Our main goal is to learn models that are able to predict spore contaminations in different types of herbs on the basis of information about the harvesting process, transport conditions, drying, and storage conditions. This shall enable us to identify optimal processing parameters, which will allow more effective and cost efficient contamination prevention. Using machine learning, we have generated ensembles of models that predict the risk for spore contamination on the basis of harvest processing parameters. The training information about contamination in herbs is given as results of laboratory analysis data. We applied different modeling algorithms (random forests, gradient boosting trees, genetic programming, and neural networks). In this paper we report on modeling results for yeast and mold contaminations in peppermint and nettle; e.g., for yeast contamination in peppermint we obtained models with 78.13% accuracy. Additionally, we use descriptive statistics to identify those parameters that have a statistically significant influence on the contamination; for example, our analysis shows that there seems to be a relationship between mold in peppermint and information about harrowing and the growth height (p = 0.001).