NADH-GOGAT Overexpression Does Not Improve Maize (Zea mays L.) Performance Even When Pyramiding with NAD-IDH, GDH and GS

Maize plants overexpressing NADH-GOGAT were produced in order to determine if boosting 2-Oxoglurate production used as a carbon skeleton for the biosynthesis of amino acids will improve plant biomass and kernel production. The NADH-GOGAT enzyme recycles glutamate and incorporates carbon skeletons into the ammonium assimilation pathway using the organic acid 2-Oxoglutarate as a substrate. Gene pyramiding was then conducted with NAD-IDH and NADH-GDH, two enzymes also involved in the synthesis of 2-Oxoglurate. NADH-GOGAT overexpression was detrimental for shoot biomass production but did not markedly affect kernel yield. Additional NAD-IDH and NADH-GDH activity did not improve plant performance. A decrease in kernel production was observed when NADH-GDH was pyramided to NADH-GOGAT and NAD-IDH. This decrease could not be restored even when additional cytosolic GS activity was present in the plants overexpressing the three enzymes producing 2-Oxoglutarate. Detailed leaf metabolic profiling of the different transgenic plants revealed that the NADH-GOGAT over-expressors were characterized by an accumulation of amino acids derived from glutamate and a decrease in the amount of carbohydrates further used to provide carbon skeletons for its synthesis. The study suggests that 2-Oxoglutarate synthesis is a key element acting at the interface of carbohydrate and amino acid metabolism and that its accumulation induces an imbalance of primary carbon and nitrogen metabolism that is detrimental for maize productivity

Evaluation of the Microbial Quality of <i>Hermetia illucens</i> Larvae for Animal Feed and Human Consumption: Study of Different Type of Rearing Substrates

In the context of climate change and depletion of natural resources, meeting the growing demand for animal feed and human food through sufficient, nutritious, safe, and affordable sources of protein is becoming a priority. The use of Hermetia illucens, the black soldier fly (BSF), has emerged as a strategy to enhance the circularity of the agri-food chain, but its microbiological safety remains a concern. The aim of the present study was to systematically review available data on the microbiological quality of BSF and to investigate the impact of using four different rearing substrates including classic options allowed by the EU regulation (cereals, fruits, vegetables) and options not allowed by EU regulations regarding vegetable agri-food (co-products, food at shelf life, and meat). A total of 13 studies were collected and synthesized, including 910 sample results, while 102 new sample results were collected from the present experiments in three farms. Both datasets combined revealed a high level of contamination of larvae, potentially transmitted through the substrate. The main pathogenic bacteria identified were Bacillus cereus, Clostridium perfringens, Cronobacter spp., Escherichia coli, Salmonella spp., and Staphylococcus aureus coagulase-positive, while Campylobacter spp. and Listeria monocytogenes were not detected. Any of these four substrates were excluded for their use in insect rearing; however, safety concerns were confirmed and must be managed by the operators of the sector using microbial inactivation treatment after the harvest of the larvae in order to propose safe products for the market. The results obtained will guide the definition of the control criteria and optimize the following manufacturing steps