A Pilot Study for the Characterization of Bacillus spp. and Analysis of Possible B. thuringiensis/Strongyloides stercoralis Correlation.

Differentiating between Bacillus species is relevant in human medicine. Bacillus thuringiensis toxins might be effective against Strongyloides stercoralis, a nematode causing relevant human morbidity. Our first objective was to evaluate genomic and MALDI-TOF identification methods for B. thuringiensis. Our secondary objective was to evaluate a possible negative selection pressure of B. thuringiensis against S. stercoralis. PCR and Sanger were compared to MALDI-TOF on a collection of 44 B. cereus group strains. B. thuringiensis toxin genes were searched on 17 stool samples from S. stercoralis-infected and uninfected dogs. Metagenomic 16S rRNA was used for microbiome composition. The inter-rate agreement between PCR, Sanger, and MALDI-TOF was 0.631 k (p-value = 6.4 × 10-10). B. thuringiensis toxins were not found in dogs' stool. Bacteroidota and Bacillota were the major phyla in the dogs' microbiome (both represented >20% of the total bacterial community). Prevotella was underrepresented in all Strongyloides-positive dogs. However, the general composition of bacterial communities was not significantly linked with S. stercoralis infection. The genomic methods allowed accurate differentiation between B. thuringiensis and B. cereus. There was no association between B. thuringiensis and S. stercoralis infection, but further studies are needed to confirm this finding. We provide the first descriptive results about bacterial fecal composition in dogs with S. stercoralis infection.

Regulatory T Cells as Predictors of Clinical Course in Hospitalised COVID-19 Patients.

Background: The host immune response has a prominent role in the progression and outcome of SARS-CoV-2 infection. Lymphopenia has been described as an important feature of SARS-CoV-2 infection and has been associated with severe disease manifestation. Lymphocyte dysregulation and hyper-inflammation have been shown to be associated with a more severe clinical course; however, a T cell subpopulation whose dysfunction correlate with disease progression has yet to be identify. Methods: We performed an immuno-phenotypic analysis of T cell sub-populations in peripheral blood from patients affected by different severity of COVID-19 (n=60) and undergoing a different clinical evolution. Clinical severity was established based on a modified WHO score considering both ventilation support and respiratory capacity (PaO2/FiO2 ratio). The ability of circulating cells at baseline to predict the probability of clinical aggravation was explored through multivariate regression analyses. Results: The immuno-phenotypic analysis performed by multi-colour flow cytometry confirmed that patients suffering from severe COVID-19 harboured significantly reduced circulating T cell subsets, especially for CD4+ T, Th1, and regulatory T cells. Peripheral T cells also correlated with parameters associated with disease severity, i.e., PaO2/FiO2 ratio and inflammation markers. CD4+ T cell subsets showed an important significant association with clinical evolution, with patients presenting markedly decreased regulatory T cells at baseline having a significantly higher risk of aggravation. Importantly, the combination of gender and regulatory T cells allowed distinguishing between improved and worsened patients with an area under the ROC curve (AUC) of 82%. Conclusions: The present study demonstrates the association between CD4+ T cell dysregulation and COVID-19 severity and progression. Our results support the importance of analysing baseline regulatory T cell levels, since they were revealed able to predict the clinical worsening during hospitalization. Regulatory T cells assessment soon after hospital admission could thus allow a better clinical stratification and patient management.

Energy efficacy used to score organic refuse pretreatment processes for hydrogen anaerobic production.

The production of hydrogen through Anaerobic Digestion (AD) has been investigated to verify the efficacy of several pretreatment processes. Three types of waste with different carbon structures have been tested to obtain an extensive representation of the behavior of the materials present in Organic Waste (OW). The following types of waste were selected: Sweet Product Residue (SPR), i.e., confectionary residue removed from the market after the expiration date, Organic Waste Market (OWM) refuse from a local fruit and vegetable market, and Coffee Seed Skin (CSS) waste from a coffee production plant. Several pretreatment processes have been applied, including physical, chemical, thermal, and ultrasonic processes and a combination of these processes. Two methods have been used for the SPR to remove the packaging, manual (SPR) and mechanical (SPRex). A pilot plant that is able to extrude the refuse to 200atm was utilized. Two parameters have been used to score the different pretreatment processes: efficiency (ξ), which takes into account the amount of energy produced in the form of hydrogen compared with the available energy embedded in the refuse, and efficacy (η), which compares the efficiency obtained using the pretreated refuse with that obtained using the untreated refuse. The best result obtained for the SPR was the basic pretreatment, with η=6.4, whereas the thermal basic pretreatment gave the highest value, η=17.0 for SPRex. The best result for the OWM was obtained through a combination of basic/thermal pretreatments with η=9.9; lastly, the CSS residue with ultrasonic pretreatment produced the highest quantity of hydrogen, η=5.2.