Calcium carbonate precipitation, a widespread phenomenon among bacteria, has been investigated due to its wide range of scientific and technological implications. Nevertheless, little is known of the molecular mechanisms by which bacteria foster calcium carbonate mineralization. In our laboratory, we are studying calcite formation byBacillus subtilis, in order to identify genes involved in the biomineralization process. A previous screening of UV mutants and of more than one thousand mutants obtained from the EuropeanB. subtilisFunctional Analysis project allowed us to isolate strains altered in the precipitation phenotype. Starting from these results, we focused our attention on a cluster of five genes (lcfA,ysiA,ysiB,etfB, andetfA) called thelcfAoperon. By insertional mutagenesis, mutant strains carrying each of the five genes were produced. All of them, with the exception of the strain carrying the mutatedlcfAoperon, were unable to form calcite crystals. By placing transcription under IPTG (isopropyl-β-d-thiogalactopyranoside) control, the last gene,etfA, was identified as essential for the precipitation process. To verify cotranscription in thelcfAoperon, reverse transcription-PCR experiments were performed and overlapping retrocotranscripts were found comprising three adjacent genes. The genes have putative functions linked to fatty acid metabolism. A link between calcium precipitation and fatty acid metabolism is suggested.

The kinetics and the metabolism of Bifidobacterium adolescentis MB 239 growing on galactooligosaccharides (GOS), lactose, galactose, and glucose were investigated. An unstructured unsegregated model for growth in batch cultures was developed, and kinetic parameters were calculated with a recursive algorithm. The growth rate and cellular yield were highest on galactose, followed by lactose and GOS, and were lowest on glucose. Lactate, acetate, and ethanol yields allowed the calculation of carbon fluxes toward fermentation products. Distributions between two- and three-carbon products were similar on all the carbohydrates (55 and 45%, respectively), but ethanol yields were different on glucose, GOS, lactose, and galactose, in decreasing order of production. Based on the stoichiometry of the fructose-6-phosphate shunt and on the carbon distribution among the products, the ATP yield was calculated. The highest yield was obtained on galactose, while the yields were 5, 8, and 25% lower on lactose, GOS, and glucose, respectively. Therefore, a correspondence among ethanol production, low ATP yields, and low biomass production was established, demonstrating that carbohydrate preferences may result from different distributions of carbon fluxes through the fermentative pathway. During the fermentation of a GOS mixture, substrate selectivity based on the degree of polymerization was exhibited, since lactose and the trisaccharide were the first to be consumed, while a delay was observed until longer oligosaccharides were utilized. Throughout the growth on both lactose and GOS, galactose accumulated in the cultural broth, suggesting that β(1-4) galactosides can be hydrolyzed before they are taken up.

Aquaculture is a globally expanding industry that contributes to feeding an increasing global population. Shellfish cultivation is one of the largest sectors of aquaculture and one of the few food productions that have the potential capacity of acting as carbon sink. In fact, >90 % of bivalve shells are calcium carbonate (CaCO₃), synthetized during biocalcification process, which incorporates a molecule of CO₂. Manila clam (Venerupis philippinarum, Adams & Reeves, 1850) and Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) are two of the major groups of cultivated shellfish. Our aim was to assess the potential role of those two bivalve species in the overall marine carbon balance using an ecosystem approach, and to evaluate if they can be definitely regarded as carbon sink. The contribution to CO₂ emissions (as CO₂ eq./kg of fresh products) due to mollusk farming has been also calculated as carbon-source term by means of Life Cycle Assessment (LCA). LCA is nowadays the most shared and accepted tool for evaluating the environmental impacts of aquaculture productions. As a case study, the Sacca di Goro coastal lagoon (Northern Adriatic Sea, Italy) has been considered, because it is the premier site in Europe for clam farming, and one of the most important for mussels. Our study has shown that for each kilogram of harvested and packaged clams and mussels, shell formation throughout the mollusk growth allows to permanently capture 254 and 146 g of CO₂, in the face of 22 and 55 g CO₂ eq. emitted for farming, respectively. As a result, clams and mussel aquaculture could be considered as a carbon sink, with a net carbon capture capacity of 233 and 91 g CO₂/kg of fresh product, respectively. In a wider context, bivalve aquaculture could be included in the carbon trading system and played a role towards the carbon-neutral economy.

Bivalves aquaculture is already considered a very sustainable for of food production and might become an essential pillar on which to develop future global food security. However, with the increase in production, a correspondingly great amount of waste will be produced all around the earth, principally in the form of shells, which can represent up to 90% of the fresh mollusk weight. Nowadays, shell waste has no notable use and is commonly regarded as waste, often dumped in landfill, or thrown back into the sea, causing a significant level of environmental concern, and resulting in a loss of natural and valuable resources. Bivalve shells are mainly formed by CaCO₃, giving them the potential to become a promising secondary raw material for several applications, from a circular economy perspective. This paper aims to review the scientific literature of the last twenty years and the principal recent trends in shell waste reuse, describing applications that are already in place on a large scale in agriculture and livestock, soils, water and wastewater remediation, biodiesel production and biofilters, as well as niche applications that now simply have the potential to be scaled up.