Spatial and Seasonal Structure of Bacterial Communities Within Alpine Vineyards: Trentino as a Case Study

Bacteria have a fundamental role in determining the fitness of grapevine, the composition of grapes and the features of wines but at present, little information is available. In this work, the bacteria colonizing the different portions of grapevine (bark, leaves and grapes) were explored in the vineyards of the Alpine region of Trentino, considering the impact of different environmental and agronomical variables. The vineyards included in the work were selected based on their different geographical positions (altitude) and grapevine training systems in order to explore the whole variability of the grapevine ecosystem. Moreover, the surface amount of copper was measured on grapes and leaves during the vegetative growth. Bacterial analysis, performed using plate counts and Illumina MiSeq, revealed an increase in the concentration of grape bacteria proportional to the progress of the ripening stage. Conversely, the peak of bacterial concentration onto leaf and bark samples occurred in August, probably due to the more favourable environmental conditions. In bark samples, the bacterial microbiota reached the 7 log CFU/cm2, while 6 log UFC/g were measured in grape samples. A remarkable biodiversity was observed, with 13 phyla, 35 classes, 55 orders, 78 families and 95 genera of bacteria present. The presence of some taxa (Alphaproteobacteria, Desulfovibrionaceae, Clostriadiales, Oscillospira, Lachnospiraceae and Bacteroidales) was ubiquitous in all vineyards, but differences in terms of relative abundance were observed according to the vegetative stage, altitude of the vineyard and training system. Bacteria having oenological implication (Lactobacillus, Pediococcus and Oenococcus) were detected in grape samples collected in August, in low abundance. The data revealed a complex bacterial ecosystem inside the vineyard that, while maintaining common traits, evolves according to environmental and agronomical inputs. This study contributes to define the role of bacteria in the complex balance established in each vineyard between human actions and agricultural environment, known as terroir.

Pilot-Scale Feasibility Study for the Stabilization of Coal Tailings via Microbially-Induced Calcite Precipitation

Sustainable long-term solutions to managing tailings storage facilities (TSFs) are integral for mines to operate in a safe and environmentally responsible manner. The long-term storage of subaqueous tailings can pose significant safety, environmental, and economic risks, therefore alternative containment strategies for maintaining geochemical stability of reactive materials must be explored. In this study, the physical and geochemical stabilization of coal tailings using microbial induced calcite precipitation (MICP) was evaluated at a laboratory pilot scale. Three application techniques simulated commonly used agricultural approaches and equipment that could be deployed for field-scale treatment: spraying on treatment solutions with irrigation sprinklers, mixing tailings and treatment solutions with a rototiller, and distributing treatment solutions via shallow trenches using an excavator ripper. Test cells containing 1.0 x 1.0 x 0.5m of tailings were treated with ureolytic bacteria (Sporosarcina pasteurii) and cementation solutions composed of urea and calcium chloride for 28 days. Penetrometer tests were performed following incubation to evaluate the extent of cementation. The spray-on application method showed the greatest strength improvement, with in an increase in surface strength of more than 50% for the 28-day testing period. The distribution of treatment solution using trenches was found to be less effective and resulted in greater variability in particle size distribution of treated tailings and would not be recommended for use in the field. The use of rototilling equipment provided a homogenous distribution of treatment solution, however, the disruption to the tailings material was less effective for facilitating effective cementation. Bacterial plate counts of soil samples indicated that S. pasteurii cultures remained viable in a tailings environment for 28 days at 18˚C and near-neutral pH. The treatment was also found to stabilize the pH of tailings porewater sampled over the 28-day incubation period, suggesting the potential for the treatment to provide short-term geochemical stability under unsaturated conditions.

Development of “Quadrello di Ovino”, a Novel Fresh Ewe’s Cheese

This work was performed to produce a new soft ewe’s milk cheese, namely “Quadrello di ovino” (QdO) cheese, to enlarge ewe’s dairy product portfolio of South Italy, barely limited to Pecorino cheese typology. Cheese making was performed applying the technology for “Crescenza” cheese typology with some modifications. In particular, pasteurized ewes’ milk was inoculated with two commercial starter formulations (SF1 and SF2) of Streptococcus thermophilus to obtain two different productions (QdO-P1 and QdO-P2, respectively). Plate counts demonstrated the ability of both starter formulations to drive the fermentation process, since S. thermophilus counts reached 109 CFU/g in both productions. Generally, the two starter formulations did not affect the chemical composition of QdO cheeses that contained, on average, 64.08% dry matter of which approximately 54.99% were fats and 36.39% proteins. Among chemical parameters, significant differences were registered for secondary lipid oxidation state (significantly lower for QdO-P2), fatty acids and volatile organic compounds (VOCs). However, the differences registered among cheese VOCs from were not perceived by the panelists who recognized both cheese productions highly similar, although QdO-P2 cheeses were mostly appreciated by the judges. This study allowed to produce a novel fresh ovine cheese with specific chemical and sensorial characteristics well appreciated by consumers.

Control of Bacillus weihenstephanensis in Pasteurized Liquid Whole Eggs Formulated with Nisin

Bacillus weihenstephanensis can grow at refrigeration temperature and cause food poisoning. It has been isolated from liquid whole egg products. The moderate heat used for pasteurization of liquid egg products is ineffective for killing spore-forming bacteria including Bacillus. Available predictive models and a pretrial study in broth suggested the potential for growth of Bacillus spp. under the tested conditions. Hence, hurdles such as storage of product below 4°C or use of preservatives would be needed to ensure the food safety of pasteurized egg products. This study evaluated the growth inhibition of B. weihenstephanensis in pasteurized liquid whole egg product formulated with 6.25 ppm nisin during storage at refrigerated and abuse refrigerated temperatures for a total 13 weeks, in three replicate trials. At day 0, the product had a pH of 7.52±0.29, while background microflora such as aerobic plate counts, presumptive B. cereus, and yeast and molds were <10 CFU/g. Product inoculated with target 2.5 log10 CFU/g of B. weihenstephanensis, stored at 4°C for 4 weeks and subsequently at 7 or 10°C for 9 weeks exhibited no growth in all three replicate trials. Average counts reduced (p<0.05) by at least one-log10 in six weeks in all samples stored at either 7 or 10°C. Similarly, growth of total plate counts, presumptive Bacillus spp., yeast and mold counts was not observed in uninoculated controls stored at 4°C for 4 weeks and subsequently at 7 or 10°C for 9 weeks. Visual and odor evaluation performed at each sampling time point showed no abnormalities. This study assessed the efficacy of maximum allowed level of nisin for use in pasteurized liquid whole eggs and validated the inhibition of B. weihenstephanensis in the product for an extended shelf life of up to 13 weeks.

Effect of Essential Oils and Vacuum Packaging on Spoilage-Causing Microorganisms of Marinated Camel Meat during Storage

The use of essential oils (EOs) and/or vacuum packaging (VP) with meats could increase product shelf-life. However, no studies investigating the effect of EOs and VP on camel meat background microbiota have been conducted previously. The study aimed to analyze the antimicrobial effect of essential oils (EOs) carvacrol (CA), cinnamaldehyde (CI), and thymol (TH) at 1 or 2% plus vacuum packaging (VP) on the growth of spoilage-causing microorganisms in marinated camel meat chunks during storage at 4 and 10 °C. VP is an effective means to control spoilage in unmarinated camel meat (CM) and marinated camel meat (MCM) compared to aerobic packaging (AP). However, after EO addition to MCM, maximum decreases in spoilage-causing microorganisms were observed under AP on day 7. Increasing the temperature from 4 to 10 °C under AP increased the rate of spoilage-causing bacterial growth in CM and MCM; however, EOs were more effective at 10 °C. At 10 °C the maximum reductions in total mesophilic plate counts, yeast and molds, mesophilic lactic Acid bacteria, Enterobacteriaceae, and Pseudomonas spp. were 1.2, 1.4, 2.1, 3.1, and 4.8 log CFU/g, respectively. Incorporating EOs at 2% in MCM, held aerobically under temperature abuse conditions, delayed spoilage.

Gold Nanorod-Decorated Metallic MoS2 Nanosheets for Synergistic Photothermal and Photodynamic Antibacterial Therapy

Light-responsive nanocomposites have become increasingly attractive in the biomedical field for antibacterial applications. Visible-light-activated metallic molybdenum disulfide nanosheets (1T-MoS2 NSs) and plasmonic gold nanorods (AuNRs) with absorption at a wavelength of 808 nm were synthesized. AuNR nanocomposites decorated onto 1T-MoS2 NSs (MoS2@AuNRs) were successfully prepared by electrostatic adsorption for phototherapy applications. Based on the photothermal effect, the solution temperature of the MoS2@AuNR nanocomposites increased from 25 to 66.7 °C after 808 nm near-infrared (NIR) laser irradiation for 10 min. For the photodynamic effect, the MoS2@AuNR nanocomposites generated reactive oxygen species (ROS) under visible light irradiation. Photothermal therapy and photodynamic therapy of MoS2@AuNRs were confirmed against E. coli by agar plate counts. Most importantly, the combination of photothermal therapy and photodynamic therapy from the MoS2@AuNR nanocomposites revealed higher antibacterial activity than photothermal or photodynamic therapy alone. The light-activated MoS2@AuNR nanocomposites exhibited a remarkable synergistic effect of photothermal therapy and photodynamic therapy, which provides an alternative approach to fight bacterial infections.

Growth of Bacillus toyonensis in Tofu Wastewater

Bacillus toyonensis has been isolated in Dumai mangrove ecosystem of Riau Province. One of factors affecting the growth of the bacteria is growth substrate. Tofu wastewater is rich in nutrition which can be used as substrate for bacterial growth. This research aimed to observe the growth of B. toyonensis in different concentration of tofu wastewater. The bacteria was grown in tofu wastewater at concentrations 8%, 10% and 12% was supplemented with 0.1 g K2HPO4, 0.15 g KH2PO4, 0.15 g NaCl and 0.5 g vitamin B12 in 100 mL distilled water. The bacterial growth was observed by using spectrophotometer at λ 610 nm and by analysis the total plate counts on plate count agar (PCA) at 0, 24, 48, 72 and 96 hour cultivation. Spectrophotometric observation showed that the highest bacterial growth of all tofu wastewater treatments indicated by the addition of 12% tofu wastewater, although the absorbance value was lower than culture in tryptic soy broth (TSB) as control. Exponential growth occurred between 0-24 hour incubation, and the highest growth indicated in substrate contained 12% tofu wastewater. Similarly, total plate count (TPC) analysis indicated that the highest bacterial growth of all treatment occurred at 24 hours incubation, and the highest count was also indicated by treatment of 12% tofu wastewater (2.42±0.06×108 CFU/mL). In conclusion, tofu wastewater can be an alternative substrate for the bacterial growth.

Optimization of Quantitative Analysis of Biofilm Cell from Pipe Materials

The quantitative analysis of biofilm can be used not only to assess the microbiological stability of tap water but also on its basis can assess: the degree of colonization of materials by bacterial cells, the rate of biofilm formation on the surfaces of pipes and determine their composition and number. The article presents the results of research on the development of an effective method of biofilm detachment from the surface of the galvanized steel. The number of biofilm cells was determined by methods: (1) luminometric ATP determination, (2) flow cytometry and (3) heterotrophic plate counts (HPC). The presence of the biofilm was confirmed by SEM and fractal analysis. The analysis of the obtained results showed that the most effective method of detaching the biofilm cells from the galvanized steel surface was the mechanical separation with a sterile cotton swab. The variant with the use of a sterile swab enables rapid collection of the biofilm from the surface of the ducts forming internal installations or water supply networks. Due to the simplicity and speed of obtaining results, the luminometric ATP measurement has been established as the best method for the quantification of biofilm cells. The results of this study were intended to provide reliable and useful data on the quantification of biofilm cells.

Antimicrobial Efficacy of Contact Lens Solutions Assessed by ISO Standards

Microbial keratitis (MK) is an eye infection caused by opportunistic bacteria or fungi, which may lead to sight-threatening corneal ulcers. These microorganisms can be introduced to the eye via improper contact lens usage or hygiene, or ineffective multipurpose solutions (MPSs) to disinfect daily wear contact lenses. Thus, the patient’s choice and use of these MPSs is a known risk factor for the development of MK. It is then critical to determine the efficacy of popular MPSs against ubiquitous ocular microorganisms. Therefore, we compare the efficacy of nine major MPSs on the global market against four different microorganism species, and with four different common contact lenses. In accordance with International Standards Organization protocol 14729 and 18259, the microorganisms were inoculated into each MPS with and without contact lenses, and held for the manufacturer’s disinfection time, 24 h, and 7 days after challenge with Serratia marcescens or Fusarium spp. Plates were incubated for 2–7 days and plate counts were conducted to determine the number of surviving microorganisms. The majority of MPSs demonstrated significantly higher disinfection efficacies without contact lenses. Broadly, among the microorganisms tested, the OPTI-FREE products (Puremoist, Express, and Replenish) maintained the highest disinfection efficacies at the manufacturer’s stated disinfection time when paired with any contact lens, compared with other MPSs. These were followed closely by RevitaLens and renu Advanced. MPSs containing dual biocides polyquaternium-1 and myristamidopropyl dimethylamine possessed the highest disinfection efficacy against multiple ocular pathogens.