Search

Preservation of cheese microbiota biodiversity is central to the sensory quality of traditional and PDO cheeses. Lyophilized commercial selected starters, being advantageous in terms of cells concentration, are supplanting natural cultures causing important loss of microbial biodiversity in the dairy environment. Biodiversity could be recovered using natural starter cultures, however their cells concentration after propagation is lower than the commercial ones. Two autochthonous and biodiverse starter cultures (MixA and MixB) coming from scotta (residual whey from Ricotta cheese manufacture), collected in the 1960 s from Pecorino Romano PDO cheese manufactures, were revitalized in reconstituted commercial powder scotta. The aim of this study was the propagation of the microbial starter mixes increasing their bacterial concentration in the pellet, reducing nonessential scotta components by a fast and not-expensive method, without changing the microbial community balance. The behaviour of each mix inoculated in scotta was compared to that in half-concentrated, clarified, and half-concentrated-clarified scotta. Higher cells concentration in the pellets from the modified scotta was obtained, without changing technological performances and microbial fingerprint. The pellets obtained were reinoculated in commercial scotta for the preparation of the scotta-innesto (the typical starter for Pecorino Romano), and no differences were observed among the treatments after incubation. The reduction of nonessential scotta's components could help the reproduction of natural starter cultures preserving their properties.

During Pecorino Romano PDO cheese production, scotta (residual whey from ricotta cheese manufacturing) or siero (whey) can be integrated with autochthonous starters, natural and composed of an indefinite number of species and strains, or commercial selected starters to obtain scotta/siero-innesto. In this study, three biodiverse autochthonous natural starter cultures (SR30, SR56, and SR63) belonging to the Agris Sardegna BNSS microbial collection, composed of different strains belonging to the species Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Enterococcus faecium, and Limosilactobacillus reuteri were lyophilized and combined into two starter mixes (A and B). The scotta/siero-innesto and the Pecorino Romano obtained using natural starters were compared with those obtained using commercial selected starters during three seasons of the cheesemaking campaign. Different pH and microbial compositions for the scotta/siero-innesto obtained using natural or commercial starters were found, attributable to their different biodiversity. The six-month-ripened cheese microbiota was influenced mostly by the season of cheesemaking, whereas physico-chemical and sensory analyses did not highlight differences among the products obtained. In general, no effect attributable to the type of scotta/siero-innesto used was observed, allowing the conclusion that natural starter cultures can be used also in industrial-scale production, ensuring high stability in the technological performances and preserving the microbial, chemical, and sensory characteristics of Pecorino Romano PDO cheese.

The effect of tetracycline (Tc) and cow manure on soil bacterial community composition and antibiotic resistance gene (ARG) abundance in soil was investigated in the present microcosm study. Effects of repeated applications of cow manure spiked with Tc in two concentrations or without Tc on the bacterial communities of a clayey and a sandy soil with different history of anthropogenic pollution by sewer flooding were investigated. Soil samples were taken 60 days after each of three amendments. Denaturing gradient gel electrophoresis (DGGE) fingerprints of 16S rRNA gene amplicons from total community DNA revealed soil type-dependent changes in the bacterial community composition in response to manure and to Tc, which became more pronounced with repeated applications. Repeated manure amendments and Tc, in particular at high concentration, triggered the further increase of ARGs tet(A), tet(O), tet(Q), tet(W), sul1, and mobile genetic elements (MGEs) IncP-1e plasmids and intI1, in a soil type-dependent manner. In the clay soil with no anthropogenic history, the ARG and MGE abundances were low or not detectable, while manure amendments caused pronounced increases in their relative abundance. In the sandy soil with a history of anthropogenic impact, ARGs and MGEs were already present at a higher level and strong increases were mainly observed for the relative abundances of sul2 and MGEs. Here, we show for the first time that effects of repeated cow manure applications might be dependent on soil type and foregoing anthropogenic soil pollution and that the presence of Tc could further increase the abundance of ARGs and MGEs.

High amounts of antibiotics are introduced in the soil environment by manure amendment, which is the most important spreading route in soil, with a potential ecotoxicological impact on the environment. The objectives of this study were (a) to assess the tetracycline (Tc) bioavailability in a clay and in a sandy soil, and (b) to evaluate the effects of the Tc and cow manure on the structure and function of soil microbial communities. Clay and sandy soils were spiked with Tc at the concentrations of 100 and 500 mg Tc kg(-1) soil, and were amended or not with cow manure. The clay soil showed greater Tc sorption capacity and bioavailable Tc was between 0.157 and 4.602 mg kg(-1) soil. Tc dose and time-dependent effects on soil microbial communities were investigated by fluorescein diacetate activity, phospholipid fatty acids analysis, as well as by Biolog community level physiological profile and microbial counts at 2, 7 and 60 days after Tc and/or manure addition. The added Tc caused detrimental effect on the microbial activity and structure, particularly in the short term at the highest concentrations. However, the Tc effect was transient' it decreased after 7 days and totally disappeared within 60 days. Cow manure shifted the bacterial structure in both soils, increased the microbial activity in clay soil and contributed to recover the microbial structure in Tc-spiked manure treatments.