NO2 air pollution drives species composition, but tree traits drive species diversity of urban epiphytic lichen communities.

Lichens serve as important bioindicators of air pollution in cities. Here, we studied the diversity of epiphytic lichens in the urban area of Munich, Bavaria, southern Germany, to determine which factors influence species composition and diversity. Lichen diversity was quantified in altogether 18 plots and within each, five deciduous trees were investigated belonging to on average three tree species (range 1-5). Of the 18 plots, two were sampled in control areas in remote areas of southern Germany. For each lichen species, frequency of occurrence was determined in 10 quadrats of 100 cm2 on the tree trunk. Moreover, the cover percentage of bryophytes was determined and used as a variable to represent potential biotic competition. We related our diversity data (species richness, Shannon index, evenness, abundance) to various environmental variables including tree traits, i.e. bark pH levels and species affiliation and air pollution data, i.e. NO2 and SO2 concentrations measured in the study plots. The SO2 levels measured in our study were generally very low, while NO2 levels were rather high in some plots. We found that the species composition of the epiphytic lichen communities was driven mainly by NO2 pollution levels and all of the most common species in our study were nitrophilous lichens. Low NO2 but high SO2 values were associated with high lichen evenness. Tree-level lichen diversity and abundance were mainly determined by tree traits, not air pollution. These results confirm that ongoing NO2 air pollution within cities is a major threat to lichen diversity, with non-nitrophilous lichens likely experiencing the greatest risk of local extinctions in urban areas in the future. Our study moreover highlights the importance of large urban green spaces for species diversity. City planners need to include large green spaces when designing urban areas, both to improve biodiversity and to promote human health and wellbeing.

Massive Survey on Bacterial-Bacteriophages Biodiversity and Quality of Natural Whey Starter Cultures in Trentingrana Cheese Production.

This study focused on the microbial and bacteriophages identification and characterization in cheese-production facilities that use natural whey starter (NWS) cultures for Trentingrana production. Bacterial and phage screening was carried out on cooked not acidified whey and NWS samples isolated from six dairy factories, for 4 consecutive days in four different months. By means of a combined approach, using plate counts, bacterial isolation, and metataxonomic analysis Lactobacillus helveticus was found occurring as the dominant species in NWS cultures and Levilactobacillus brevis as codominant in the cheese factories where the temperature of NWS production was mainly lower than 40°C, suggesting that the variability in the parameters of the NWS culture preparation could differently modulate the bacterial species in NWS cultures. Using turbidity test approach on 303 bacterial isolates from the NWS cultures, 120 distinct phages were identified. L. helveticus phage contamination of NWS cultures was revealed in most of the analyzed samples, but despite the great recovery of bacteriophage contamination cases, the microbial quality of NWS cultures was high. Our results support the presence of natural bacteriophage resistance mechanisms in L. helveticus. The use of NWS cultures probably creates an ideal environment for the proliferation of different L. helveticus strains balanced with their phages without a clear dominance. It is evident, from this study, that the presence of a high biodiversity of NWS bacterial strains is relevant to avoid phages dominance in NWS cultures and consequently to keep a good acidification ability.

Raw Milk Microbiota Modifications as Affected by Chlorine Usage for Cleaning Procedures: The Trentingrana PDO Case.

Milk microbiota represents a key point in raw milk cheese production and contributes to the development of typical flavor and texture for each type of cheese. The aim of the present study was to evaluate the influence of chlorine products usage for cleaning and sanitizing the milking equipment on (i) raw milk microbiota; (ii) the deriving whey-starter microbiota; and (iii) Trentingrana Protected Designation of Origin (PDO) cheese microbiota and volatilome. Milk samples from three farms affiliated to a Trentingrana PDO cheese factory were collected three times per week during a 6-weeks period in which a sodium hypochlorite detergent (period C) was used and during a subsequent 6-weeks period of non-chlorine detergent usage (period NC). Samples were subjected to microbiological [Standard Plate Count; coliforms; coagulase-positive staphylococci; and lactic acid bacteria (LAB)] and metagenomic analysis (amplification of V3-V4 regions of 16S rRNA gene performed on Illumina MiSeq platform). In addition, cheese volatilome was determined by SPME-GC-MS. In the transition from period C to period NC, higher SPC and LAB counts in milk were recorded. Milk metagenomic analysis showed a peculiar distinctive microbiota composition for the three farms during the whole experimental period. Moreover, differences were highlighted comparing C and NC periods in each farm. A difference in microbial population related to chlorine usage in bulk milk and vat samples was evidenced. Moreover, chlorine utilization at farm level was found to affect the whey-starter population: the usually predominant Lactobacillus helveticus was significantly reduced during NC period, whereas Lactobacillus delbrueckii had the exact opposite trend. Alpha- and beta-diversity revealed a separation between the two treatment periods with a higher presence of L. helveticus, L. delbrueckii, and Streptococcus thermophilus in cheese samples after NC detergent period. Cheese volatilome analysis showed a slight decrease in lipolysis during C period in the inner part of the cheese wheel. Although preliminary, these results suggest a profound influence on milk and cheese microbiota, as well as on raw milk cheese production and quality, due to the use of chlorine. However, further studies will be needed to better understand the complex relationship between chlorine and microbiota along all the cheese production steps.