Pilot scale study on UV-C inactivation of bacterial endospores and virus particles in whole milk: evaluation of system efficiency and product quality

UV-C processing of whole milk (WM) using a designed pilot scale Dean flow system was conducted at flow rates (11.88, 23.77, and 47.55 gph), Reynolds number ranges from 2890-11562 and the Dean number (at curved region) calculated as (648-2595) to inactivate bacterial endospores and virus particles. Biodosimetry studies were conducted to quantify the reduction equivalent fluence at selected experimental conditions. Results revealed that the dose distribution improved as flow rate increases, attributed to increase in Dean effects and turbulence intensity. Microbial inactivation studies conducted at 47.55 gph showed 0.91 (stdev:0.15) and 2.14 (stdev:0.19) log reduction/ pass for B. cereus endospores and T1UV phage. Linear inactivation trend was observed against number of passes which clearly demonstrates equivalent dose delivery during each pass. Lipid peroxidation value and volatile profile did not change significantly at UV dose of 60 mJ/cm 2. Lower E EO value signifies the higher electrical efficiency of the system.

Thermal resistance and high-performance microwave decontamination assessment of Bacillus endospores isolated from food-grade herbal extracts

Generally, endospore contamination can occur from different sources during product manufacturing in many industries and therefore lower its quality by affecting physicochemical properties and shelf-life. Bacterial endospores can germinate inside the product and produce several enzymes, which can cause several undesirable changes. This study assessed the spores thermal resistance and applied a microwave decontamination technique toward herbal extracts (Tilia tomentosa and Centella asiatica) containing ethanol or glycerol. Based on 16S rRNA analysis, the detected contaminant endospores belonged to different Bacillus species, namely B. subtilis, B. zhangzhouensis, and B. pumilus. The thermal resistance assessment using inoculated endospores in the actual products revealed B. pumilus T2 as the most resistant endospore to the heat treatments tested in both T. tomentosa and C. asiatica extracts. Finally, a high-performance microwave technique was used to decontaminate T. tomentosa extract against the mixture of Bacillus spores. Results from the microwave technique indicate that the increase of temperature from 100°C to 105°C not only decontaminated the product but also could dramatically decrease the effective thermal treatment time (10 times), which can benefit the product quality. The results provided in this study considerably contribute to improving an original decontamination method for products containing glycerol and ethanol with the most negligible effect on product quality.

Impact of Heat Treatment on the Microbiological Quality of Frass Originating from Black Soldier Fly Larvae (Hermetia illucens)

Since black soldier fly larvae (BSFL, Hermetia illucens) are being produced at substantial volumes, concomitantly large amounts of the resulting by-product, called frass, are generated. This frass can potentially be applied as valuable plant fertilizer or soil improver. Since frass carries high microbial counts, potentially including foodborne pathogens, safety problems for consumers should be prevented. A heat treatment of 70 °C for 60 min is proposed to reduce harmful organisms in insect frass, based on EU regulations ((EU) No. 2021/1925). This study evaluated for the first time the impact of the proposed heat treatment on BSFL frass. This was done by applying the treatment on uninoculated frass as well as on frass inoculated with Salmonella or Clostridium perfringens at 5.0 log cfu/g. The heat treatment resulted in a reduction (maximum one log-cycle) of total viable counts and did not noticeably reduce bacterial endospores. In contrast, Enterobacteriaceae counts were reduced to below the detection limit (10 cfu/g). Heat treatment of inoculated frass resulted in absence of Salmonella in 25 g of frass and reduction of vegetative C. perfringens to below the detection limit (1 cfu/g). The proposed heat treatment appears to be appropriate to meet the microbiological regulations for insect frass.

Geological processes mediate a subsurface microbial loop in the deep biosphere

The deep biosphere is the largest microbial habitat on Earth and features abundant bacterial endospores1,2. Whereas dormancy and survival at theoretical energy minima are hallmarks of subsurface microbial populations3, the roles of fundamental ecological processes like dispersal and selection in these environments are poorly understood4. Here we combine geophysics, geochemistry, microbiology and genomics to investigate biogeography in the subsurface, focusing on bacterial endospores in a deep-sea setting characterized by thermogenic hydrocarbon seepage. Thermophilic endospores in permanently cold seabed sediments above petroleum seep conduits were correlated with the presence of hydrocarbons, revealing geofluid-facilitated cell migration pathways originating in deep oil reservoirs. Genomes of thermophilic bacteria highlight adaptations to life in anoxic petroleum systems and reveal that these dormant populations are closely related to oil reservoir microbiomes from around the world. After transport out of the subsurface and into the deep-sea, thermophilic endospores re-enter the geosphere by sedimentation. Viable thermophilic endospores spanning the top several metres of the seabed correspond with total endospore counts that are similar to or exceed the global average. Burial of dormant cells enables their environmental selection in sedimentary formations where new petroleum systems establish, completing a geological microbial loop that circulates living biomass in and out of the deep biosphere.

Potential of Fermentation and Vacuum Packaging Followed by Chilling to Preserve Black Soldier Fly Larvae (Hermetia illucens)

Black soldier fly larvae (Hermetia illucens) are currently reared at an industrial scale, mainly as a feed ingredient. The logistic chain not only involves the production of larvae, but also stabilisation, storage, and transport. The aim of this work was to study fermentation and vacuum packaging of larvae as potential preservation technologies. For fermentation, blanched larvae were pulverised into a paste, and a starter culture, NaCl, and glucose were added. The mixture was fermented for 7 days at 35 °C and then stored for 14 days at 4 °C and pH and microbial counts were monitored. Vacuum packaging was applied to living, blanched and frozen larvae. After packaging, they were stored for 6–10 days at several temperatures and gas composition, survival (living larvae) and microbial counts (killed larvae) were recorded. Fermentation allows storage of pulverised larvae, but points to consider are a rapid pH reduction and the presence of bacterial endospores. Vacuum packaging did not bring added value over cooling alone. This was the case for all types of larvae investigated. Vacuum packaging is not considered as a valuable preservation technology to pursue for storage and transport of black soldier fly larvae.

Effect of dietary supplementation with powder derived from Moringa oleifera and Azadirachta indica leaves on growth and microbial load of edible crickets

Cricket farming has shown its potential to address food and nutrition insecurity in parts of the world. However, one of the remaining challenges limiting mass production of edible insects is the affordability of feeds. One possible way to address this issue would be to explore the use of traditional plants such as Moringa oleifera leaf (MOL) and Azadirachta indica leaf (AIL) that are locally available, are protein-rich and possesses some antibacterial properties in insect feed formulation. In this regard, a study was conducted to evaluate the effect of supplementation of MOL (5 or 10%) and AIL (5 or 10%) powder in the commercially used starter chicken feed on growth and microbial load of house cricket (Acheta domesticus) and field cricket (Gryllus bimaculatus). After feeding for four weeks, the supplemented feeds with 10% MOL or 10% AIL in A. domesticus and G. bimaculatus significantly decreased (P<0.05) the body weights. Also, supplementation with MOL (5 or 10%) and AIL (5 or 10%) increased the mortality in both cricket species. Although high counts of total aerobic, Enterobacteriaceae, lactic acid bacteria, bacterial endospores, yeasts and moulds were observed, significantly lower counts (P<0.05) of lactic acid bacteria and bacterial endospores were observed in treatments containing MOL (5 or 10%) and 10% AIL in both cricket species. After processing (toasting and boiling), significant reductions (P<0.001) of all vegetative microbial cells were observed but bacterial endospores were not completely eliminated. Thus, during production and processing of insects and insect-derived products, bacterial endospores would require special attention. Salmonella and Escherichia coli were not detected in processed crickets’ samples. In conclusion, supplementation with 5% MOL or 5% AIL can produce crickets with similar body weight as control feed. Mortality increased significantly when feeds were supplemented with either MOL or AIL powder. Supplementation with MOL and AIL powders significantly reduced the cell numbers for some bacterial groups in both cricket species.

Synergistic Effects of Nisin, Lysozyme, Lactic Acid, and CitricidalTM for Enhancing Pressure-Based Inactivation of Bacillus amyloliquefaciens, Geobacillus stearothermophilus, and Bacillus atrophaeus Endospores

The inactivation of bacterial endospores continues to be the main curtailment for further adoption of high-pressure processing in intrastate, interstate, and global food commerce. The current study investigated the effects of elevated hydrostatic pressure for the inactivation of endospore suspension of three indicator spore-forming bacteria of concern to the food industry. Additionally, the effects of four bacteriocin/bactericidal compounds were studied for augmenting the decontamination efficacy of the treatment. Elevated hydrostatic pressure at 650 MPa and at 50 °C was applied for 0 min (untreated control) and for 3, 7, and 11 min with and without 50K IU of nisin, 224 mg/L lysozyme, 1% lactic acid, and 1% CitricidalTM. The results were statistically analyzed using Tukey- and Dunnett’s-adjusted ANOVA. Under the condition of our experiments, we observed that a well-designed pressure treatment synergized with mild heat and bacteriocin/bactericidal compounds could reduce up to >4 logs CFU/mL (i.e., >99.99%) of bacterial endospores. Additions of nisin and lysozyme were able, to a great extent, to augment (p < 0.05) the decontamination efficacy of pressure-based treatments against Bacillus amyloliquefaciens and Bacillus atrophaeus, while exhibiting no added benefit (p ≥ 0.05) for reducing endospores of Geobacillus stearothermophilus. The addition of lactic acid, however, was efficacious for augmenting the pressure-based reduction of bacterial endospores of the three microorganisms.

Visible Light Driven Photocatalytic Decolorization and Disinfection of Water Employing Reduced TiO2 Nanopowders

Defect-engineering of TiO2 can have a major impact on its photocatalytic properties for the degradation of persisting and non-biodegradable pollutants. Herein, a series of intrinsic and extrinsic defects are induced by post annealing of crystalline TiO2 under different reducing atmospheres. A detailed optoelectronic characterization sheds light on the key characteristics of the defect-engineered TiO2 nanopowders that are linked to the photocatalytic performance of the prepared photocatalysts. The photodegradation of a model dye, malachite green, as well as the inactivation of bacterial endospores of the Geobacillus stearothermophilus species were studied in the presence of the developed catalysts under visible light illumination. Our results indicate that a combination of certain defects is necessary for the improvement of the photocatalytic process for water purification and disinfection under visible light.