Maximum likelihood estimation (MLE) is a popular method for parameter estimation in both applied probability and statistics but MLE cannot solve the problem of incomplete data or hidden data because it is impossible to maximize likelihood function from hidden data. Expectation maximum (EM) algorithm is a powerful mathematical tool for solving this problem if there is a relationship between hidden data and observed data. Such hinting relationship is specified by a mapping from hidden data to observed data or by a joint probability between hidden data and observed data. In other words, the relationship helps us know hidden data by surveying observed data. The essential ideology of EM is to maximize the expectation of likelihood function over observed data based on the hinting relationship instead of maximizing directly the likelihood function of hidden data. Pioneers in EM algorithm proved its convergence. As a result, EM algorithm produces parameter estimators as well as MLE does. This tutorial aims to provide explanations of EM algorithm in order to help researchers comprehend it. Moreover some improvements of EM algorithm are also proposed in the tutorial such as combination of EM and third-order convergence Newton-Raphson process, combination of EM and gradient descent method, and combination of EM and particle swarm optimization (PSO) algorithm.

Since the mid-1990s probiotics have been proven to support the treatment of several gastrointestinal diseases. Additionally, probiotics have been taken as a common panacea for civilization diseases. To date, the only probiotic yeasts with evidence of health-promoting effects are Saccharomyces cerevisiae var. boulardii. The probiotic properties are strain-dependent, whereby it is crucial to develop methods to verify the probiotic-enriched food authentication including the growing number of dietary supplements with simplified commercialisation procedures. Due to the significant genetic similarity of S. cerevisiae var. boulardii to S. cerevisiae, qPCR-HRM analysis was tested, which has a very high sensitivity for polymorphism detection and enables simultaneous identification of the microorganism in the presence of the reference sample. The effectiveness of interspecies primer pairs and intragenus primer pairs designed to amplify heterogeneous regions was examined. HRM analysis using primers for 18SrRNA and ITS allowed for identification of S. cerevisiae var. boulardii at the species level. S. cerevisiae var. boulardii was efficiently identified at the variety level with amplification of HO and RPB2 in the single- and multi-component DNA extracts. The RPB2 amplicon showed the highest intraspecies differentiation power. The predominance of S. cerevisiae var. boulardii in the mixtures tested was crucial for identification using qPCR-HRM analysis and quantitatively corresponded to the strain content in the probiotic-enriched food matrices.

Kininogens are multidomain glycoproteins found in the blood of most vertebrates. They are important in the blood coagulation cascade pathways - in intrinsic pathway activation leading to thrombin generation partially independently from tissue factor dependent extrinsic pathway, connecting blood coagulation with the kallikrein-kinin system. Nothing is known about the shape on atomic level therefore the endeavor to obtain the good-quality spatial structure of kininogens is important for a better understanding of their role in disease and treatment. Application of cryo-EM is important for solving the spatial structure of kininogens, drawing new frontiers in understanding the function and opening new pathways for drug development.

Centrifugal pumps are widely employed in the oil refinery industry due to their efficiency and effectiveness in fluid transfer applications. The reliability of pumps plays a pivotal role in ensuring uninterrupted plant productivity and safe operations. Analysis of failure history data shows that bearings have been identified as critical components in oil refinery pump groups. However, traditional reliability estimation theories may not apply when data is limited or subject to right censoring. This paper addresses the complexity of estimating the Weibull distribution parameters using the maximum-likelihood method under the abovementioned conditions. The likelihood equation lacks an explicit analytical solution, necessitating the use of numerical methods for resolution. The proposed approach presented in this article leverages the Expectation-Maximization (EM) algorithm for estimating the Weibull distribution parameters. This method provides more accurate estimates of failure rates and probabilities by accounting for limited and censored data. The findings are demonstrated through a case study, showcasing the practical application of the proposed approach.

Despite well-documented health benefits of probiotics Saccharomyces, its application in oral health has not been comprehensively assessed. Dental caries is a transmissible disease initiated by acid production of cariogenic bacteria and yeast, such as Streptococcus mutans and Candida albicans, on tooth enamel and followed by subsequent enamel demineralization. Here, we investigated the effect of two Saccharomyces strains (Saccharomyces boulardii, Saccharomyces cerevisiae) on S. mutans-C. albicans cross-kingdom interactions using a cariogenic planktonic model. Viable cells, pH changes and gene expression were measured. S. cerevisiae and S. boulardii inhibited the growth of C. albicans either in dual-, or multi-species conditions. Saccharomyces also inhibited C. albicans hyphal formation. Furthermore, Saccharomyces reduced the acidity of the culture medium which usually plummeted below pH 5 when S. mutans and C. albicans were present in the model. The presence of Saccharomyces maintained the culture medium above 6 even after overnight incubation, demonstrating a protective potential against dental enamel demineralization. S. boulardii significantly downregulated S. mutans atpD and eno genes expression. Overall, our results shed light on a new promising candidate, Saccharomyces, for dental caries prevention due to its potential to create a less cariogenic environment marked by a neutral pH and reduced growth of C. albicans.

Gelatin industry residues are increasingly used as fertilizer and soil conditioner. However, correct residue dosage is critical for grass development and minimizing environmental impacts. This randomized block design study determined adequate dosage of Mixed Gelatin Sludge (MGS) for Marandu grass production in wet/dry seasons in Brazil. Five MGS levels (0%–200% of required nitrogen) were compared to mineral fertilizer. Agronomic/productivity characteristics, bromatological composition, macro/micro-nutrient composition of leaves, and soil chemical attributes were evaluated. Agronomic/productivity characteristics were influenced by MGS dose in both dry/rainy seasons, except for leaf blade pseudostem ratio and percentage of leaves/pseudostem. Bromatological composition was influenced by MGS doses in dry/rainy seasons except for dry/mineral material quantities. Marandu leaf tissue chemical composition was significantly influenced by MGS dose, except for potassium, boron, and iron. Chemical composition of four soil layers between 0–50 cm influenced GMS dose, except for pH, organic matter, magnesium, copper, manganese, and zinc. GMS dose for Marandu production should be 200% of nitrogen (N) requirement. GMS application increased productivity/quality of Marandu grass. Macronutrients (N, phosphorus) and micronutrients (Ca, Mg, S, Cu, and Zn) increased in Marandu grass and in the soil (Ca, S, and Na). Increased sodium (Na) level was not limiting.

Developing recombinant Saccharomyces cerevisiae strains capable of transporting and fermenting cellobiose directly is a promising strategy for second-generation ethanol production from lignocellulosic biomass. In this study we cloned and expressed in the CEN.PK2-1C strain an intracellular βglucosidase (SpBGL7) from Spathaspora passalidarum, and co-expressed the cellobiose transporter SiHXT2.4 from Scheffersomyces illinoinensis, and two putative transporters from Candida tropicalis (CtCBT1) and Meyerozyma guilliermondii (MgCBT2). While all three transporters allowed cell growth on cellobiose, only the MgCBT2 permease allowed cellobiose fermentation, although cellobiose consumption was stuck (incomplete). The analysis of the βglucosidase and transport activities revealed that the cells stopped to consume cellobiose due to a drop in the transport activity. Since ubiquitinylation of lysine residues at the N- or C-terminal domains of the permease are involved in the endocytosis and degradation of sugar transporters, we constructed truncated versions of the permease lacking lysine residues at the C-terminal domain (MgCBT2ΔC), and at both the C- and N-terminal domain (MgCBT2ΔNΔC), and co-expressed these permeases with the SpBGL7 βglucosidase in an industrial strain. While the strain harboring the MgCBT2ΔC transporter continued to produce stuck cellobiose fermentations as the wild-type MgCBT2 permease, the strain with the MgCBT2ΔNΔC permease was able to consume and ferment all the cellobiose present in the medium. Thus, our results highlight the importance of expressing cellobiose transporters lacking lysine at the N- and C-terminal domains for efficient cellobiose fermentation by recombinant S. cerevisiae.

The unicellular eukaryote S. cerevisiae is an invaluable resource for the study of basic eukaryotic cellular and molecular processes. However, due to its small size compared to other eukaryotic organisms the study of subcellular structures is challenging. Expansion microscopy (ExM) holds great potential to study the intracellular architecture of yeast, especially when paired with pan-labelling techniques visualising the full protein content inside cells. ExM allows to increase imaging resolution by physically enlarging a fixed sample that is embedded and cross- linked to a swellable gel followed by isotropic expansion in water. The cell wall present in fungi – including yeast – and Gram-positive bacteria is a resilient structure that resists denaturation and conventional digestion processes usually used in ExM protocols, resulting in uneven expansion. Thus, the digestion of the cell wall while maintaining the structure of the resulting protoplasts are crucial steps to ensure isotropic expansion. For this reason, specific experimental strategies are needed, and only a few protocols are currently available. We have developed a modified ExM protocol for S. cerevisiae, with 4x expansion factor, which allows the visualisation of the ultrastructure of the cells. Here, we describe the experimental procedure in detail, focusing on the most critical steps required to achieve isotropic expansion for ExM of S. cerevisiae.

Saccharomyces cerevisiae (S. cerevisiae) ethanol fermentation has left an indelible trace in human history. It is regarded as the original beverage synthesis, and has developed into an essential energy supply for mankind today. However, the history of ethanol synthesis using S. cerevisiae, including the mechanism analysis process, has not been carefully and succinctly reported. In this paper, we review the history of ethanol synthesis using S. cerevisiae. In addition, we also review the substrates that drive S. cerevisiae ethanol synthesis, including the utilization of glucose, sucrose, starch and molasses, in order to summarize the current development status and to contribute to the further improvement of this field.

Chondroitin is a glycosaminoglycan that has gained widespread use in nutraceuticals and pharmaceuticals, mainly for treating osteoarthritis. Traditionally, it has been extracted from animal cartilage but recently, biotechnological processes have emerged as a commercial alternative to avoid the risk of viral or prion contamination and offer a vegan-friendly source. Typically, these methods involve producing the chondroitin backbone using pathogenic bacteria and then modifying it enzymatically through the action of sulfotransferases. Despite the challenges of expressing active sulfotransferases in bacteria, the use of eukaryotic microorganisms is still limited to a few works using Pichia pastoris. To create a safer and efficient biotechnological platform, we have constructed a biosynthetic pathway for chondroitin production in S. cerevisiae as a proof-of-concept. Up to 125 mg/L and 200 mg/L of intracellular and extracellular chondroitin were produced, respectively. Furthermore, as genome-scale models are valuable tools for identifying novel targets for metabolic engineering, a stoichiometric model of chondroitin-producing S. cerevisiae has been developed and used in optimization algorithms. Our research has yielded several novel targets, such as uridine diphosphate(UDP)-N-acetylglucosamine pyrophosphorylase (QRI1), glucosamine-6-phosphate acetyltransferase (GNA1) or N-acetylglucosamine-phosphate mutase (PCM1) overexpression that might enhance chondroitin production, which will guide future experimental research to develop more efficient host organisms for the biotechnological production process.

Yeast extract is widely used in different food industries as a flavoring agent or vitamin supplement. In this study, a process was optimized for the production of yeast extract from Baker’s yeast (Saccharomyces cerevisiae). A glass vessel stirred fermenter was used for the cultivation of yeast biomass. The effect of various physical and chemical factors was evaluated on the production of yeast cells and optimum conditions for the production of maximum yeast biomass were determined. The optimum growth was obtained at 30ºC with pH 4.5 using molasses as a substrate supplemented with urea at 150rpm. Yeast cells were then separated by centrifugation and ruptured and autolysis was observed to be the most feasible method. Among various method employed to dry the yeast extract, spray dryer appeared as most efficient one. Yeast extract obtained after drying was subjected to different analyses and compared with commercial yeast extract. The produced yeast extract was applied in media preparation to grow different microorganisms including yeast, bacteria and fungi and considerable growth was observed. These results indicated that the developed process is a cost effective alternate approach for the production of yeast extract.

This study aimed to evaluate the capacity of Saccharomyces cerevisiae for Cadmium absorption in Milk. Nowadays one of the most serious problems is heavy metals pollution. Applying microorgaisms as a novel biotechnology is so useful especially in foodstuffs. Among the biosorbents for heavy metals’ removal, Saccharomyces cerevisiae has got an increasing attention due to its popularity in food industry. In this regard, the effects of some important factors such as the initial metal concentration, biomass concentration and contact time on the biosorption capacity of Saccharomyces cerevisiae were studied. The biosorption was analyzed by the inductively coupled plasma mass spectrometer (ICP-MS). The maximum Cd bioremoval (70%) was at 80 μg/L of this metal concentration in milk samples containing 30×10⁸ CFU Saccharomyces cerevisiae at the end of storage time (the 4^th day). There were no significant differences in sensory and physicochemical properties of milk samples during storage (p < 0.05). The isotherm studies followed by two popular models; Langmuir and Freundlich and the results showed a better fit to the Langmuir isotherm. All together, the results of this project demonstrated that the approach of using this valuable yeast, could be applied for food and drinks’ detoxification and producing healthier foods.

Consumer demands for new sensory experiences have driven the research of unconventional yeasts in beer. While much research exists on the use of various common Saccharomyces cerevisiae strains as well as non-Saccharomyces yeasts, there exists a gap in knowledge regarding other non-cerevisiae Saccharomyces species in the fermentation of beer, outside that of S. pastorianus. Here, five distinct species of Saccharomyces from the UC Davis Phaff Yeast Culture Collection, as well as one interspecies hybrid from Fermentis, were chosen to ferment 40 L pilot scale beers. S. kudriavzevii, S. mikatae, S. paradoxus, S. bayanus, and S. uvarum yeasts were fermented in duplicate, with one fermenter in each pair receiving 10 g/L dry-hop during fermentation. Analytical measurements were made each day of fermentation and compared to controls of SafAle US-05 and SafLager W 34/70 for commercial brewing parameters of interest. Finished beers were also analyzed for aroma, taste, and mouthfeel to determine the flavor of each yeast as it pertains to brewing potential. All beers exhibited spicy characteristics, likely from the presence of phenols; dry-hopping increased fruit notes while also increasing perceived bitterness and astringency. All of the species in this study displayed great brewing potential, and might be an ideal addition to beer depending on a brewery’s desire to experiment with flavor and willingness to bring a new yeast into their production environment

Genome-scale engineering and custom synthetic genomes are reshaping the next generation of industrial yeast strains. The Cre-recombinase mediated chromosomal rearrangement mechanism of designer synthetic Saccharomyces cerevisiae chromosomes, known as SCRaMbLE, is a powerful tool which allows rapid genome evolution upon command. This system is able to generate millions of novel genomes with potential valuable phenotypes, but the excessive loss of essential genes often results in poor growth or even the death of cells with useful phenotypes. In this study we expanded the versatility of SCRaMbLE to industrial strains, and evaluated different control measures to optimise genomic rearrangement, whilst limiting cell death. To achieve this, we have developed RED (Rapid Evolution Detection), a simple colorimetric plate-assay procedure to rapidly quantify the degree of genomic rearrangements within a post-SCRaMbLE yeast population. RED-enabled semi-synthetic strains were mated with haploid progeny of industrial yeast strains to produce stress tolerant heterozygous diploid strains. Analysis of these heterozygous strains with the RED-assay, genome sequencing and custom bioinformatics scripts demonstrated a correlation between RED-assay frequencies and physical genomic rearrangements. Here we show that RED is a fast and effective method to evaluate optimal SCRaMbLE induction times of different Cre-recombinse expression systems for the development of industrial strains.

The yeast Saccharomyces cerevisiae is widely used in food and nonfood industries. During industrial fermentations yeast are exposed to fluctuations in oxygen concentration, osmotic pressure, pH, ethanol concentration, nutrient availability and temperature. Fermentation performance depends on the ability of different yeast strains to adapt to these changes. Suboptimal growth conditions trigger responses to these external stimuli to allow cellular homeostasis to be maintained. Stress-specific signaling pathways are activated to coordinate changes in tran-scription, translation, protein function, and metabolic fluxes while a transient arrest of growth and cell cycle progression occur. cAMP-PKA, HOG-MAPK and CWI signalling pathways are signal transduction pathways turned on during stress re-sponse. Comprehension of the mechanisms involved in the responses and in the adaptation to these stresses during fermentation is key to improving this industrial process. The scope of this review is to outline the advancement of knowledge about the cAMP-PKA signalling and the crosstalk of this pathway with the CWI and HOG-MAPK cascades in response to the environmental challenges heat and hy-perosmotic stress.

The aim of the present trial was to evaluate the growth performance of buffalo calves fed diets characterized by different forage/concentrate ratio with or without the Saccharomyces Cerevisiae supplementation (CBS 493.94, Yea-Sacc®). Twenty-four male buffalo calves (mean age 145.1 ± 16.1 days; mean weight 108.0 ± 18.7) were assigned randomly to 4 groups, homogeneous for age, fed four different diets: diet 1 F:C ratio 50:50; diet 2 F:C ratio 30:70; diet 3 F:C ratio 50:50 + Yea-Sacc® and diet 4 F:C ratio 30:70 + Yea-Sacc®. Buffalo calves were individually weighted before the start of the experiment and the data were used as a covariate, monthly until the end of the trial. Dry matter intake (DMI), daily weight gain (DWG) and feed conversion ratio (FCR) were calculated. The differences in diets’ composition significantly (p

This study aimed to evaluate the effect of three yeast strains, Pichia guilliermondii (Levica 27), Candida norvegensis (Levazoot 15), and a commercial product (Levucell® SC 10) containing Saccharomyces cerevisiae, on in vitro rumen fermentation of two substrates: corn stover and a total mixed ration (TMR). Samples were taken 24 hours post-fermentation to measure pH, ammonia nitrogen (NH3-N), gas production, and volatile fatty acid (VFA) concentrations. The addition of yeasts did not affect gas production at 24 hours for either diet. For pH, differences between yeast strains were noted when TMR was used. The lowest NH3-N concentration (6.1 mM) was recorded with corn stover, showing similar results across strains. Conversely, NH3-N levels with TMR ranged from 47 mM to 66 mM. The inclusion of all three yeast strains increased the molar concentrations of total VFA, as well as acetic (C2), propionic (C3), and butyric (C4) acids in the rumen fluid. The C2:C3 ratio remained unchanged with corn stover but decreased with the inclusion of Levica 27 and Levazoot 15 in the TMR. With the commercial product Levucell® SC 10, this ratio was comparable to the control treatment. In conclusion, Levica 27 and Levazoot 15 demonstrated a stimulatory effect on in vitro rumen fermentation for the evaluated diets, evidenced by increased VFA concentrations.

The yeast Saccharomyces cerevisiae ensures successful fermentation in winemaking, although the persistent use of commercial strains lead the loss of aroma complexity of wines. Hence, the research of indigenous S. cerevisiae with proper oenological features and well adapted to specific wine-growing area become of great interest for winemakers. Here, 206 S. cerevisiae strains were isolated from two wineries during two-year sampling campaign and bio-typed with the aim to evaluate the occurrence and persistence of S. cerevisiae wild population linked to each winery. Both wineries belonging to the same Verdicchio DOC wine area (Castelli di Jesi), and never used commercial yeasts during fermentation. Results showed a specific and varied population of S. cerevisiae in each winery without cross-contamination each other and with commercial starter strains. Moreover, inside each winery it was observed a persistent of some dominant biotypes over the time, indicating a sort of “winery-effect”. The evaluation of S. cerevisiae populations for the oenological characters showed a proper and well distinct aromatic imprinting on the resulted wines confirming the “winery effect”.

In this study is developed an aerobic and anaerobic stoichiometric model for Saccharomyces cerevisiae, compartmentalized in mitochondria and cytosol. This model considers the central metabolism of S. cerevisiae and it possesses the peculiarity of having catabolic and anabolic biochemical reactions for the synthesis of the higher alcohols contained in tequila; involving 94 metabolites and 117 reactions; of which 93 correspond to biochemical internal reactions and 24 to transport fluxes between the medium and the cell. The model is validated under aerobic and anaerobic conditions for the main fermentation metabolites and it coincides with experimental results and those in silico reported in the literature. This model is used to obtain three different physiological states of S. cerevisiae through of estimation of its distributions of fluxes calculated from experimental data reported in literature of fermentation in continuous culture during the tequila production under different dilution rate (0.04-0.12 h^-1). The model developed constitutes a tool for the estimation of flux distribution maps during fermentation processes for the production of tequila, which could permit estimate yields and visualize different fermentation scenarios.

Conjugation is a bacterial mechanism for DNA transfer from a donor cell to a wide range of recipients, including both prokaryotic and eukaryotic cells. In contrast to conventional DNA delivery techniques, such as electroporation and chemical transformation, conjugation eliminates the need for DNA extraction, thereby preventing DNA damage during isolation. While most established conjugation protocols allow for DNA transfer in liquid media or on a solid surface, we developed a procedure for conjugation within solid media. Such a protocol may expand conjugation as a tool for DNA transfer to species that require semi-solid or solid media for growth. Conjugation within solid media could also provide a more stable microenvironment in which the conjugative pilus can establish and maintain contact with recipient cells for the successful delivery of plasmid DNA. Furthermore, transfer in solid media may enhance the ability to transfer plasmids and chromosomes greater than 100 kbp. Using our optimized method, plasmids of varying sizes were tested for transfer from E. coli to S. cerevisiae. We demonstrated that there was no substantial decrease in conjugation frequency as plasmid size increased—up to 138.5 kbp in length. Finally, we established an efficient PCR-based synthesis protocol to generate custom conjugative plasmids

Concern for global health security and the environment due to the emergence of antibiotic-resistant bacteria and antibiotic residues in meat and other livestock products has led many countries to restrict the use of antibiotics in animal feed. This experiment was performed to assess the impact of dietary supplementation with a probiotic (Bacillus subtilis) and a postbiotic (Saccharomyces cerevisiae fermentation product) on growth performance, carcass traits, blood haemato-biochemical profile, gut microbiome, gut morphology, and immune response in broilers as an alternative to antimicrobials in poultry production system to minimize the effect on global health security. A total of 324-day-old Ven Cobb 400 broiler chicks were randomly divided into three dietary groups, each containing 12 replicated pens (n = 12), .and each replicate contained nine broiler chickens. The dietary groups consisted of 1) a basal diet without any growth promoting (T1), 2) the basal diet augmented with Bacillus subtilis at 200 g/MT feed (T2), and 3) the basal diet supplemented with Saccharomyces cerevisiae fermentation product at 1.25 kg/MT feed (T3). To calculate body weight gain, all birds and residual feed were weighed on a replicated basis over days 0, 7, 14, 21, 28, 35, and 42; mortality was recorded daily. Each replicate's mortality percentage was calculated and used to adjust BW, ADFI, and FCR calculations at the trial's end. At the end of the trial (42 d), two chickens from each replicate (24 chickens per group) were slaughtered for carcass traits, gut microbiome, and morphology measurements. Blood samples were collected for the haemato-biochemical profile on 35d and antibody titer on 28d and 35d.Feeding with SCFP (T3 group) significantly improved average daily feed intake (ADFI) and average daily gain (ADG) of chickens compared to the T1 (control) and T2 (probiotic) groups from 1 to 14 days of age. FCR was significantly improved (P<0.05) in SCFP-fed birds (T3) relative to the control (T1)over the entire experimental period (1-42 days). Carcass traits and blood haemato-biochemical parameters remained unaffected (P>0.05) by any diets. However, cholesterol levels and concentrations of corticosterone were significantly lower (P<0.05) in SCFP-fed birds (T3) compared to those of the probiotic (T2) and control (T1) groups. Total E. coli, Enterohaemorrhagic E. coli, ESBL-producing Enterobacteriaceae, and Salmonella counts were significantly higher in the T1group compared to T3 and T2 groups (P<0.05) and Salmonella counts were lower in T3 when compared to T2 (P<0.05). However, there was no significant difference in Lactobacillus count among treatment groups. PCR based characterization of ESBL- producing Enterobacteriaceae revealed maximum presence of blaCTX-M-Type (6/15, 40%), followed by blaSHV-Type (5/15, 33.3%) and blaTEM-Type (4/15, 26.6%). No variation in the occurrence pattern of ESBL/beta-lactamase associated genes in the studied ESBL- producing Enterobacteriaceae isolates was observed between the treatment groups. A significant increase in villi height and villi height to crypt depth ratio (VH: CD) (P<0.05) was observed in both T3 and T2 groups. Contrarily, villi widths did not vary across the dietary groups (P>0.05). The control group (T1) displayed a notable deeper in crypts when compared to the T3 and T2 groups (P<0.05). On day 28, the SCFP-fed birds (T3) and those fed the probiotics (T2) exhibited a significant increase (P<0.05) in antibody titers against Newcastle disease virus and Infectious bursal disease virus. Increase in antibody titre against IBDV in birds with the addition of SCFP in the diet was detected for the first time. However, there were no significant differences observed on day 35. The phagocytic activity of neutrophils and lymphocytes in broiler chickens did not show any noticeable variations among the different groups at day 35(P>0.05).It can be concluded that Saccharomyces cerevisiae fermentation product and Bacillus subtilis probiotic could be viable alternatives to antimicrobials in poultry production considering beneficial impacts in broilers fed antibiotic-free diet.

Saccharomyces cerevisiae fermentation products (SCFP) can potentially promote gastrointestinal growth and immunity in young livestock. However, reports within the existing literature on the effects of SCFP supplements on calves have been inconsistent; therefore, we perform the following experiments to resolve the inconsistencies. A total of 22 Holstein calves [10 d after birth, BW = 48.93 ± 3.99 kg (mean ± SD)] were assigned randomly into two groups, namely the control group (CON) and SCFP group, each having 11 replicas. The calves in the CON were fed a basal diet, while the SCFP group was fed the basal diet supplemented with 5g/head/d SCFP (NutriTek, Diamond V, Cedar Rapids, IA52404, United States) incorporated into feed. All the calves were regularly fed thrice daily at 08:00, 14:30, and 21:00 and had free access to water. A 5-day adaptation phase was followed by a 45-day experimental period. The results showed that compared to the CON, at the end of the d-45 trial, the body weight was significantly greater in the SCFP group (p < 0.05), and during the 1-45 days, the ADG was higher (p < 0.05). The FCR in the 30-45 days SCFP group was higher (p < 0.05). Furthermore, the apparent digestibility of DM, CP, EE, ADF, Ca, and P were significantly increased in the SCFP group, except for NDF (p < 0.05). The concentration of GH and IGF-1 in serum showed a tremendous increase (p < 0.05) with SCFP supplementation on d 15 and d 45. On d 15, SCFP supplementation significantly increased the serum IgA contents (p < 0.05). Notably, on d 15 and d 45, the serum concentrations of IL-1β, IL-6, and TNF-α reduced (p < 0.001). Moreover, the Actinobacteriota in the SCFP group were significantly lower than those in the CON group (p = 0.034). SCFP significantly increased the abundance of Butyricimonas, Parabacteroides, and Ruminococcus. The differences among sob, Chao1, and PD-tree groups were statistically significant (p < 0.05). In conclusion, these results demonstrate that SCFP supplementation improved ADG, apparent digestibility and serum hormone, enhanced immunity, and regulated gut microbiota, thereby jointly promoting the growth of pre-weaning calves.

In Brazil, sucrose-rich broths (cane juice and/or molasses) are used to produce billions of liters of both fuel ethanol and cachaça per year using selected Saccharomyces cerevisiae industrial strains. Aiming to improve this disaccharide fermentation, we have first determined the genetic characteristics, by array comparative genomic hybridization, of a group of 8 fuel ethanol and 5 cachaça industrial yeasts that tend to dominate the fermentors during the production season. The widespread presence of SUC genes encoding invertase at multiple telomeres has been shown to be a common feature of both baker’s and distillers’ yeast strains and is postulated to be an adaptation to sucrose-rich broths. Our results show that only two strains (one fuel ethanol and one cachaça yeast) have amplification of the SUC genes, with high invertase activity. The other industrial yeast strains had a single SUC locus (SUC2) in their genome, with different patterns of invertase activity. These results indicate that invertase activity probably does not limit sucrose fermentation during fuel ethanol and cachaça production by these industrial strains. Using this knowledge, we changed the mode of sucrose metabolism of an industrial strain by avoiding extracellular invertase activity, overexpressing the intracellular invertase, and increasing its transport through the AGT1 permease. This approach allowed higher ethanol production from sucrose by the modified industrial yeast when compared to its parental strain.

The aim of this study was to isolate and characterize autochthonous Saccharomyces cerevisiae yeasts from Cabernet Sauvignon grape must and to analyze how they impacted the final characteristics of the wines obtained from the same must type. This work was carried out in a “Pago” winery located in Requena, Spain. Twenty-two isolates were obtained from grape must. After molecular identification and typing, 11 different strains were determined. Growth-related parameters and metabolic characteristics (glucose and fructose consumption, and ethanol, glycerol and acetic acid production) were determined by laboratory-scale fermentations (50 mL) of Cabernet Sauvignon must. From 2 L fermentation batches, residual sugar, density, ethanol concentration, titratable and volatile acidities, pH, and polyphenolic and volatile compounds were determined, as well as the sensory attributes of the produced wines. Differences between strains in the growth and metabolic parameters were observed. Eleven S. cerevisiae strain profiles were characterized to establish which strain or strains of the selected yeasts would produce the highest polyphenolic and aromatic concentration of the red Cabernet wines made in the “Pago winery” to then multiply them for their use to make the red wines of this winery.

The Dph1•Dph2 heterodimer from yeast is a radical SAM (RS) enzyme that generates the 3-amino-3-carboxy-propyl (ACP) precursor for diphthamide, a clinically relevant modification on eukaryotic elongation factor 2 (eEF2). ACP formation requires SAM cleavage and atypical Cys-bound Fe-S clusters in each Dph1 and Dph2 subunit. Intriguingly, the first Cys residue in each motif locates next to another, ill-defined cysteine that we show in-here is conserved across eukaryotes. As judged from structural modeling, the orientation of these tandem cysteine motifs (TCM) suggests a candidate Fe-S cluster ligand role. Hence, we generated by site-directed DPH1 and DPH2 mutagenesis, Dph1•Dph2 variants with cysteines from each TCM replaced individually or in combination by serines. Assays diagnostic for diphthamide formation in vivo reveal that while single substitutions in the TCM of Dph2 cause mild defects, double mutations almost entirely inactivate the RS enzyme. Based on enhanced Dph1 and Dph2 subunit instability in response to cycloheximide chases, the variants with Cys substitutions in their cofactor motifs are particularly prone to protein degradation. In sum, we identify a fourth functionally cooperative Cys residue within the Fe-S motif of Dph2 and show that the Cys-based cofactor binding motifs in Dph1 and Dph2 are critical for the structural integrity of the dimeric RS enzyme in vivo.

Reversed Flow Gas Chromatography (R.F.G.C.) was employed to assess the impact of genetic modification on Saccharomyces cerevisiae yeast strains during the process of alcoholic fermentation, utilizing fig syrup. Multiple fermentations were carried out at various temperatures to evaluate the influence of genetic modifications on yeast strain efficiency. The study involved a wild-type yeast strain, W303, as a control, and two genetically modified strains, W_M4_533 and W_M4_558, sharing the same genetic background as the wild type. Notably, the genetic modifications in the Msn4p transcription factor involved the substitution of serine residues with alanine at positions 533 and 558, resulting in the development of psychrophilic or ethanol-resistant strains. Utilizing the R.F.G.C. method enabled the differentiation of the duration of alcoholic fermentation phases, providing insights correlated to the yeast cell life cycle. The values of rate constants (k) for each phase, conducted with both wild-type and genetically modified cells using RFGC, aligned with existing literature. Additionally, the calculation of activation energies for distinct phases revealed lower values for genetically modified strains compared to wild-type strains. This decrease in activation energies suggests enhanced efficiency in the alcoholic fermentation process for the genetically modified strains.

This study evaluated the anti-inflammatory effects, protection of gut barrier integrity, and stimulation of phagocytosis in peripheral cells of a nutritional supplement based on a synergistic combination of yeast-based ingredients with a unique 1,3/1,6-glucan complex and a consortium of postbiotic Saccharomyces cerevisiae rich in selenium and zinc. The anti-inflammatory effect in Caco-2 cells in the presence and absence of a pro-inflammatory challenge (tumour necrosis factor alpha [TNF-α]/interferon gamma [IFN-ɣ]) showed statistically significant reductions of IFN-ɣ induced protein-10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) levels vs. controls (p &lt; 0.001). Disruption of the gut integrity in the presence or absence of Escherichia coli (ETEC H10407) showed transepithelial electrical resistance (TEER) values higher in the ABB C1® group after 6 hours of testing. Spontaneous build-up of the gut epithelium monolayer over 22 days was also greater in the ABB C1® condition vs. a negative control. ABB C1® showed a significantly higher capacity to stimulate phagocytosis as compared with controls of algae β-1,3-glucan and yeast β-1,3/1,6 glucan (p &lt; 0.001). This study supports the mechanism of action by which ABB C1® may improve the immune response and be useful to prevent infection and allergy in clinical practice.

Antifungal properties are found in essential oil extracts of oregano (Origanum vulgare), rosemary (Salvia rosmarinus), clove (Syzygium aromaticum), thyme (Thymus), cinnamon (Cinnamomum verum), basil (Ocimum basilicum), therefore, the ability to prevent or inhibit the growth of Cladosporium spp., Lasiodiplodia spp., Colletotrichum spp., Fusarium spp., and Aspergillus spp. was studied. Cladosporium spp., Lasiodiplodia spp. Colletotrichum spp. Fusarium spp. y Aspergillus spp. fungi that were isolated and purified from the rot found on the banana peels (Musa paradisiaca). To perform a visual (macro-scopic) analysis of the Petri dish containing the pure fungus and a more detailed (microscopic) analysis of the fungal spores to classify it by genus, then, dilutions are prepared to inoculate onto amended PDA medium with chloramphenicol. Weekly re-inoculations are performed until iso-lated and purified fungi are obtained, which are then evaluated for in vitro inhibition and in vivo growth, according to the results of the in vivo analysis, the fungi were classified from higher to lower severity in: Colletotrichum spp., Lasiodiplodia spp., Aspergillus spp., Fusarium spp. and Cladosporium spp and based on fungal growth observations, it was found that basil and rosemary do not inhibit growth at the maximum concentration. Cinnamon inhibits at 400 ppm, rosemary halts fungal growth at 600 ppm, clove at 1000 ppm, and oregano at 800 ppm.

Leishmaniasis represents a severe global health problem. In the last decades, there have been significant challenges in controlling this disease due to the unavailability of licensed vaccines, the high toxicity of the available drugs, and an unrestrained surge of drug-resistant parasites, and human immunodeficiency virus (HIV)–Leishmania co-infections. Leishmania spp. preferentially invade macrophage lineage cells of vertebrates for replication after subverting cellular functions of humans and other mammals. These early events in host–parasite interactions are likely to influence the future course of the disease. Thus, there is a continuing need to discover a simple cellular model that reproduces the in vivo pathogenesis. Acanthamoeba spp. are non-mammalian phagocytic amoeba with remarkable similarity to the cellular and functional aspects of macrophages. We aimed to assess whether the similarity reported between macrophages and Acanthamoeba spp. is sufficient to reproduce the infectivity of Leishmania spp. Herein, we analyzed co-cultures of Acanthamoeba castellanii or Acanthamoeba polyphaga with Leishmania infantum, Leishmania amazonensis, Leishmania major, and Leishmania braziliensis. Light and fluorescence microscopy revealed that the flagellated promastigotes attach to the A. castellanii and/or A. polyphaga in a bipolar and or random manner, which initiates their uptake via pseudopods. Once inside the cells, the promastigotes undergo significant changes, which result in the obligatory amastigote-like intracellular form. There was a productive infection with a continuous increase in intracellular parasites. However, we frequently observed intracellular amastigotes in vacuoles, phagolysosomes, and the cytosol of Acanthamoeba spp. Our findings corroborate that Leishmania spp. infects Acanthamoeba spp. and replicates in them but does not cause their rapid degeneration or lysis. Overall, the evidence presented here confirms that Acanthamoeba spp. have all prerequisites and can help elucidate how Leishmania spp. infect mammalian cells. Future work exposing the mechanisms of these interactions should yield novel insights into how these pathogens exploit amoebae.

This paper discusses how mathematical models are used to predict environmental conditions. While these models are successful for larger scale phenomena, it becomes more complex for local weather or wave processes. Several approaches have been proposed to address this problem, including neural networks and statistical and mathematical models. The novelty in this approach is the use of statistical manifolds, which provide a more accurate way to estimate distances and cost functions. This work focuses on advancements in differential and information geometry for creating different types of models that can be used for environmental simulation and forecasting. These fields are vital for many institutes and centres worldwide, as they help solve challenges in areas such as meteorology, renewable energy, and aviation safety. Some potential IG applications to advance energy were given. Closing remarks with new open problems combined with the next research phase are provided.

The opportunistic pathogen Pseudomonas aeruginosa is responsible for numerous chronic infections and has a case fatality rate of 50%. All P. aeruginosa strains express β–lactamase, an enzyme capable of inactivating β–lactam antibiotics, including penicillins. However, β–lactams are still considered desirable therapeutics by physicians due to their low toxicity. The short-term goal of this study was to create an optimal model for ampC expression in P. aeruginosa and determine the selecting antibiotic to be used during mutagenesis. In the long term, this study aims to renew effectiveness of β–lactams by identifying potential allosteric binding sites on β–lactamase for the development of novel β–lactamase inhibitors. To create a system for ampC expression, the chromosomal P. aeruginosa ampC gene encoding for β–lactamase was cloned into Escherichia coli, then conjugated into P. aeruginosa ∆ampC. β– lactamase assays on recombinant E. coli and P. aeruginosa detected much higher production of β–lactamase in pMMB67HE, which was confirmed with antibiotic susceptibility testing (AST) to numerous β-lactams. From AST, carbenicillin was chosen as the antibiotic to select functional β–lactamase production during mutagenesis. Future work includes conducting mutagenesis on ampC to identify allosteric binding sites on the β–lactamase enzyme that may enable the discovery of new β–lactamase inhibitors. This work allows for the development of novel β–lactamase inhibitors that could renew the utility of β–lactams and drastically reduce morbidity and mortality from chronic Pseudomonas infections.

Wine is a processed beverage that offers high nutritional and health benefits. It is produced from grape must, which undergoes fermentation by yeasts (and sometimes lactic acid bacteria) to create a product that is highly appreciated by consumers worldwide. However, if only one type of yeast, specifically Saccharomyces cerevisiae, were used in the fermentation process, the resulting wine would lack aroma and flavor and may be rejected by consumers. To produce wine with a desirable taste and aroma, non-Saccharomyces yeasts are necessary. These yeasts contribute volatile aromatic compounds that significantly impact the wine’s final taste. They promote the release of primary aromatic compounds through a sequential hydrolysis mechanism involving several glycosidases unique to these yeasts. This review will discuss the unique characteristics of these yeasts (Schizosaccharomyces pombe, Pichia kluyveri, Torulaspora delbrueckii, Wickerhamomyces anomalus, Metschnikowia pulcherrima, Hanseniaspora vineae, Lachancea thermotolerans, Candida stellata, and others) and their impact on wine fermentations and co-fermentations. Their existence and the metabolites they produce enhance the complexity of wine flavor, resulting in a more enjoyable drinking experience.

Background: Aflatoxins (AFs) as toxic compounds are widely spread. They are frequent contaminants of crops that are produced by fungi species. Corn is of crops that widely used in the corn dry milling process; a four-step procedure comprises grain handling and milling, liquefaction and saccharification, fermentation, and co-product recovery that produces ethanol and animal feed. However, it is possible that corn as a feed for dry milling process might be contaminated with AFs. Therefore, monitoring AFs in during widespread this process results having a better understanding of the fate of AFs. Methods: In this study, a sample of corn kernel was contaminated with a high level of AFs using Aspergillus parasiticus. Then, the corn flour comprising 23400.5 µg/kg of the total level of aflatoxins (AFT), was hydrolyzed by enzymes and acid for ethanol fermentation with Saccharomyces cerevisiae. Also, the level of AFs was monitored during ethanol fermentation process. Results: The results showed that AFs did not detect in the ethanol produced through fermentation and it was safe. Also, during the experiments, the AFT level reduced by 58.5 and 95.7% for the ethanol fermentation with simultaneous saccharification and fermentation and ethanol production with acidic hydrolysis, respectively. Conclusion: The results of this study showed that highly contaminated corn with AFs can be used for ethanol fermentation through dry milling process and the produced ethanol was safe.

A dataset collected from Central-South Finland is used to investigate the mortality of boreal trees. The mortality rate is found to be the order of three times that predicted by earlier Nordic mortality models, being in the upper range of international literature observations. Small subpopulations of any tree species tend to die out. Mortality of downy birch increases with stand basal area, as well as with stand age. Mortality of Norway spruce and silver birch increases after 100 years, while that of Scots pine is invariant to age. As the relative loss rate of basal area is insensitive to stand basal area, the mortality of trees does not strongly regulate thinning practices.

The increasing demand for craft beer drives the search for novel ale yeast cultures from brew-ing-related wild environments. The focus of bioprospecting for craft cultures is to identify feral yeasts suitable to imprint unique sensorial attributes to the final product. Here, we integrated phylogenetic, genotypic, genetic, and metabolomic techniques to demonstrate that sour beer during aging in wooden barrel is a source of suitable craft ale yeast candidates. Differently from traditional lambic beer maturation phase, during aging of sour-matured production style beer, different biotypes of Saccharomyces cerevisiae dominated the cultivable in-house mycobiota, fol-lowed by Pichia membranifaciens, Brettanomyces bruxellensis, and Brettanomyces anomalus. In addi-tion, three putative S. cerevisiae x Saccharomyces uvarum hybrids were identified. S. cerevisiae feral strains sporulated, produced viable monosporic progenies, and have STA1 gene as well as the full-length promoter. During hopped wort fermentation four S. cerevisiae strains and the S. cere-visiae x S. uvarum hybrid WY213 overcame non-Saccharomyces strains in fermentative rate and ethanol production, except for P. membranifaciens WY122. This strain consumed maltose after a long lag phase, in contrast with the phenotypic profile descripted for the species. According to STA1+ genotype S. cerevisiae partially consumed dextrin, but no 4-vynil guaiacol was detected among VOCs, suggesting that these strains were phenolic off flavor-negative (POF-). In conclu-sion, the strains characterized here have relevant brewing properties and are exploitable as in-digenous craft beer starters.

In recent years, humanity has faced a growing challenge posed by the rise of antibiotic-resistant bacteria, resulting in an alarming surge in fatalities and the emergence of increasingly hard-to-manage diseases. Among these resilient pathogens is Acinetobacter baumannii, due to its escalating prevalence in hospitals, elevated mortality rates, and resistance to treatment. Despite its clinical significance, our understanding of this pathogen in non-hospital settings and its virulence factors remains limited. Therefore, this review aims to shed light on the latest research regarding its ecological niche, microbiological traits, and antibiotic resistance profiles. Recent investigations have expanded its detection to various environmental niches, including rivers, treatment plants, and soils. Moreover, this bacterium has been discovered in diverse food sources such as meat and vegetables, as well as in farm animals and household pets like dogs and cats. This broader presence of Acinetobacter baumannii outside of hospital environments underscores a significant risk of environmental contamination, necessitating heightened awareness and preventive measures to address potential public health implications.

Green spaces localized in cities create favourable environmental conditions for Ixodes ricinus, the most widespread and important vector of tick-borne pathogens in Central Europe. In this study, we described the prevalence of Borrelia burgdorferi s.l. and Babesia species found in mono- and double infections among I. ricinus ticks occurring in urban areas of the city of Poznań, west-central Poland. Two tick groups were examined: (i) a group of 1,029 host-seeking ticks (460 nymphs, 289 females, and 280 males and (ii) a group of 1,268 engorged female ticks removed from 1,115 tick-infested animals, dogs and cats. The overall prevalence of B. burgdorferi s.l. found in host-seeking ticks was almost two-fold higher in comparison to ticks derived from pets (8.7 vs. 4.7%, respectively, χ2 test, P = 0.001). Borrelia afzelii and B. garinii were the most prevalent species both in ticks from vegetation (3.7% and 3.7%, respectively) and from pets (3.7% and 0.6%, respectively). Babesia infections were slightly more common in feeding ticks compared to those from vegetation (6.0 vs. 4.7%, respectively), however, the difference was not significant. Three Babesia species including Ba. microti, Ba. venatorum and Ba. canis were identified. Babesia canis and Ba. microti were the most prevalent in host-seeking (2.6% and 1.4%, respectively) and feeding ticks (2.8% and 2.2%, respectively), whereas Ba. venatorum was the rarest piroplasm. All Ba microti obtained sequences (n=42) were identical to Ba. microti genotype "Jena/Germany" that is considered as pathogenic to humans. Furthermore, the sequences of Ba. venatorum (n=17) were identical with those isolated from patients in Italy, Austria and Poland. Altogether, out of 121 Babesia-positive ticks, 19.8% were simultaneously infected with B. burgdorferi s.l. spirochetes. We found similar prevalences of co-infections in host-seeking (1.1%) as well as in feeding ticks (1.0%). In the first group, Ba. canis prevailed in coinfections (73%), mostly with B. afzelii and B. garinii, in the second group, Babesia microti dominated with B. afzelii (92.7%). This is the first finding documenting the co-occurrence of both pathogens in ticks from companion animals. The detection of Ba. microti and Ba. venatorum sequences identical to strains pathogenic to humans as well as predominance of LB spirochetes, B. afzelii and B. garinii in I. ricinus from vegetation and animals indicates that there is a potential risk of acquiring human babesiosis and/or Lyme borreliosis in urban areas of the city of Poznań. Although, the overall risk of encountering a Ba. microti and B. burgdorferi s.l. co-infected tick was low (1.0%), the potential for more severe clinical symptoms and complications deserves notice. A relatively high infection rates of Ba. canis in dog-derived ticks (3.2%) and in host-seeking ticks (2.6%), suggests that I. ricinus could potentially be involved in the circulation of this piroplasm in areas, where the competent vector, Dermacentor reticulatus is absent or rare.

Pagiophloeus tsushimanus, a new record of wood-boring pest first reported in China in 2014, has significantly infested Cinnamomum camphora across various administrative districts in Shanghai. And it shows a concerning trend of spread. The larval stage of the insect, being concealed within the trunks of C. camphora, exhibits characteristics such as hidden activity, strong destructiveness, and difficulty in control. This severely affects the normal growth of the host trees. While the primary host plant for P. tsushimanus is the C. camphora, preliminary observations have shown that the pest can also complete its life cycle on Cinnamomum chekiangensis and Phoebe chekiangensis. Both of them are significant species for urban greening. To explore the host selection preference of this beetle, this study investigates the olfactory selection behavior of adult P. tsushimanus towards C. camphora, C. chekiangensis, and P. chekiangensis. Results from choice tests, no-choice tests, and "Y"-shaped olfactometer experiments indicated that both male and female adults exhibited a feeding preference for C. camphora branches, with females showing a preference for laying eggs on C. camphora branches as well. Behavioral responses demonstrated a positive chemotaxis towards volatiles from C. camphora and a negative chemotaxis towards volatiles from C. chekiangensis and P. chekiangensis. Overall, the findings suggest that both male and female adults have a selective preference for volatiles emitted by C. camphora. These provides a theoretical basis for monitoring and controlling host shift occurrences in P. tsushimanus.

The fate of critically sick patients with infections in intensive care units (ICUs) is significantly influenced by antimicrobial resistance, which has become a serious concern in developing nations. This observational study was conducted at the IBN Sina Specialised Hospital in Dhaka, Bangladesh, over a 12-month period from January 2021 to December 2021, in order to track the pattern of antibiotic resistance among the patients admitted to the intensive care unit (ICU). Aspiration pneumonia (29%) and diabetes mellitus (24%), among 200 patients, were the most common main diagnoses. 65 samples (or 26%) of the 250 samples had 85 bacteria identified from them. Urine (14.12%) and tracheal aspirate (64.71%) were the most frequently found sites of infection. In the sample from the tracheal aspirate, Pseudomonas spp. (30.59%), Escherichia coli (24.71%), Acinetobacter spp. (20%), and Klebsiella spp. (14.12) were the most frequently isolated microorganisms. The following antibiotics were shown to have the highest overall patterns of resistance: levofloxacin (68.24), amikacin (64.71), meropenem (49.41), ceftazidime (75.29), ciprofloxacin (78.82), and gentamicin (82.35). The most effective antibiotic against Klebsiella species was meropenem (64.61%), while Acinetobacter species were mostly susceptible to cotrimoxazole (64.67%) and piperacillin + tazobactam (60.50%). Most susceptible to netilmicin (70.48%) and meropenem (49.32%) were Escherichia coli, while Pseudomonas spp. were mostly responsive to colistin (55.14%) and netilmicin (52.25%). Antibiotic resistance is common among intensive care unit (ICU) patients, and most isolated microbes have resistance to traditional medicines.

A three-year survey was conducted to estimate the incidence of grapevine trunk diseases (GTDs) in Greece and identify fungi associated with the disease complex. In total, 310 vineyards in different geographical regions in northern, central, and southern Greece were surveyed, and 533 fungal strains were isolated from diseased vines. Morphological, physiological and molecular (5.8S rRNA gene-ITS sequencing) analyses revealed that isolates belonged to 35 distinct fungal genera, including well-known (e.g. Botryosphaeria sp, Diaporthe spp., Eutypa sp., Diplodia sp., Fomitiporia sp., Phaeoacremonium spp., Phaeomoniella sp.) and lesser-known (e.g. Neosetophoma sp., Seimatosporium sp., Didymosphaeria sp., Kalmusia sp.) grapevine wood inhabitants. GTDs-inducing population structure differed significantly among the discrete geographical zones. Phaeomoniella chlamydospora (26.62%, n=70), Diaporthe spp. (18.25%, n=48) and F. mediterranea (10.27%, n=27) were the most prevalent in Heraklion, whereas D. seriata, Alternaria spp., P. chlamydospora and Fusarium spp. were predominant in Nemea (central Greece). In Amyntaio and Kavala (northern Greece), D. seriata was the most frequently isolated species (&gt;50% frequency). Multi-genes (rDNA-ITS, LSU, tef1-α, tub2, act) sequencing of selected isolates, followed by pathogenicity tests revealed that Neosetophoma italica, Seimatosporium vitis, Didymosphaeria variabile and Kalmusia variispora caused wood infection with the former being the most virulent. To the best of our knowledge, this is the first report of N. italica associated with GTDs worldwide. This is also the first record of K. variispora, S. vitis and D. variabile associated with wood infection of grapevine in Greece. The potential associations of disease indices with vine age, cultivar, GTD-associated population structure and the prevailed meteorological conditions in different viticultural zones in Greece are presented and discussed.

This research explores the predictive capabilities of autonomous driving systems by integrating human factor considerations within the context of safety-critical events. Recognizing the significance of human behaviors in influencing driving dynamics, the study employs advanced modeling techniques to enhance the accuracy of predictions in scenarios that demand heightened safety measures. Traditional rule-based systems and monotonic logic often fall short in addressing the complexities of safety-critical events. To overcome these limitations, the research proposes the application of non-monotonic logic, allowing for flexible and adaptive reasoning that accommodates exceptions and context-specific information.The study emphasizes the importance of incorporating individual differences among drivers, such as risk-taking tendencies, reaction times, decision-making processes, and driving styles. By considering these human factors, the research aims to develop realistic and accurate autonomous driving models that capture the nuances of real-world driving scenarios, especially in safety-critical situations. The predictive model takes into account both internal and external factors, enabling the autonomous system to anticipate and respond effectively to unforeseen events.The primary goal is to provide autonomous vehicles with the capability to make plausible inferences, handle conflicting data, and adapt their behavior in real-time during safety-critical events. The proposed model integrates personalized cognitive agents for each driver, incorporating their unique preferences, characteristics, and needs. This personalized approach aims to optimize the safety and efficiency of autonomous driving, contributing to the ongoing evolution of intelligent transportation systems.In conclusion, this research contributes to advancing the field of autonomous driving by introducing a predictive model that leverages human factor considerations to enhance safety in safety-critical events. The incorporation of non-monotonic logic and individualized cognitive agents signifies a comprehensive approach to address the challenges associated with predicting autonomous driving behavior, paving the way for safer and more reliable autonomous vehicles in dynamic and unpredictable environments.

Background: The bacterium Capnocytophaga spp. is a part of the oral microbiome of dogs, which has zoonotic potential. This study aimed to assess factors that influence the prevalence of Capnocytophaga cynodegmi and Capnocytophaga canimorsus in Wrocaw’s dog population. Methods: Molecular detection (PCR) involved 105 swabs, examining parameters such as oral cavity condition, underlying health issues, recent antibiotics, age, medications, material collection, and dog chews. Results: The study highlighted significant associations between the presence of Capnocytophaga cynodegmi and oral cavity condition, material collection method, and use of dog’s chews. Conclusions: Correlations obtained in this study between the oral health of Wroclaw's dog population and the presence of Capnocytophaga cynodegmi suggest its significance in the oral microbiome of dogs. Limitations, including the small sample size, highlight the need for more research to comprehensively understand additional factors that affect the prevalence of Capnocytophaga spp. in dogs

Boswellia trees are the source of frankincense, which can be found throughout the Middle East and parts of Africa and Asia. Boswellia serrata extract has been shown to have anti-cancer, anti-inflammatory, and anti-microbial effects. Periodontitis is an oral chronic inflammatory disease that affects nearly half of the US population. We investigated the antimicrobial effects of B. serrata extract on two oral pathogens associated with periodontitis. Using the minimum inhibitory concentration and crystal violet staining methods, we demonstrated that Porphyromonas gingivalis growth and biofilm formation were impaired by treatment with B. serrata extracts. However, there were no significant effects on Fusobacterium nucleatum growth and biofilm formation. Using colony-forming units quantification and microscopy techniques, we also showed that concentrations of B. serrata that were not toxic for host cells decreased intracellular P. gingivalis infection in human gingival epithelial cells. Our results show antimicrobial activity of a natural product extracted from Boswellia trees (B. serrata) against periodontopathogens. Thus, B. serrata has the therapeutic potential for preventing and/or treating periodontal diseases. Future studies will identify molecular components of B. serrata extracts responsible for the beneficial effects.

Celtis sinensis is an adaptable species that is widely grown in southern China. In June 2022, a leaf blotch disease of C. sinensis was observed in Nanjing, Jiangsu, China. Based on morphological characterization, three isolates were identified as Alternaria species. Phylogenetic analysis of combined ITS, GAPDH, TEF1-α, RPB2 and Alt a 1 sequences identified the three isolates we obtained as Alternaria alternata and Alternaria koreana. Koch’s postulates were fulfilled in the greenhouse, and the pathogenicity of A. alternata and A. koreana was determined by leaf inoculation tests on C. sinensis seedlings. The symptoms of indoor inoculation were consistent with those in the field. A. alternata and A. koreana can grow at 15-35 °C, with an optimal growth temperature of 25 °C. The results of fungicide sensitivity experiments indicated that A. alternata and A. koreana were the most sensitive to prochloraz, which may be a useful strategy for the future prevention and control management of A. alternata and A. koreana. This study provides the first step for further research on A. alternata and A. koreana as pathogens of C. sinensis and provides a theoretical basis for future control strategies.

This study focuses on examining various approaches for Tensor Decompositions (TDs) and their potential applications in satellite imaging (SI) and deep learning (DL). The research highlights how these decompositions can contribute to the advancement of SI and DL, providing valuable insights for future utilization of TDs in research. It explores how these techniques contribute to the advancement of SI and DL methods, providing insights into their potential applications and suggesting future research directions. The study aims to enhance the use of TDs techniques to further advance research efforts in these fields. More importantly, the current investigation offers a thorough analysis of the potential advantages and reasons for employing TDs techniques in different domains, including satellite imaging and deep learning. The aim is to enhance research outcomes by utilizing TDs. The review also identifies unresolved issues and proposes future directions for further investigation in this field. Fundamentally, these proposed open problems will open new grounds to the research community to articulate, innovate, and provide more real-life applications to improve the current state of the art by delving into a wider vision for a higher-level performance of both satellite imaging (SI) and deep learning (DL). Looking at the bigger scenario, this also suggests that TDs could be potentially employed to revolutionize existing machine learning technologies as well as the current space AI industry.

It was aimed evaluate the nutritive value of three foods (Cecropia sp., Pterodon sp., and Inga sp.), for sloths (Bradypus variegatus), based on in vitro gas production. After a 14-day adaptation period to these foods, approximately 500g of gastric contents were collected from three female sloths, processed, and incubated with food samples to evaluate digestibility and in vitro degradation kinetics. The neutral detergent fiber (NDFcp) was greater (P&lt;0,05) in the leaves of Cecropia sp. The non-fibrous carbohydrate contents were greater in Pterodon sp. The greatest cellulose content was found in the leaves of C. pachystachya, as well as lowest value for hemicellulose. There were differences in the in vitro digestibility of crude protein (P&lt;0.05), higher for Inga sp. In terms of kinetic parameters, Pterodon sp. demonstrated higher production of total gases (Vt) and digestion rates of fibrous carbohydrates (kdNFC) (P&lt;0.05), possibly related to the lower NDF content and higher values of soluble carbohydrates, which favored fermentative processes. The leaves of Pterodon sp. and Inga sp. demonstrated potential for feeding B. variegatus. Feeding B. variegatus with Cecropia sp., due to the NDF content, can cause negative effects on dry matter consumption.

In the Ebro Delta, the abundance of burrowing bivalves has dramatically decreased, with the blue crab, Callinectes sapidus, being blamed by shellfish collectors. Trends from 2010 evidence a decrease in the capture of clams (Ruditapes spp.) before 2016 (start of blue crab fisheries), although further decline of both clam and cockle (Cerastoderma glaucum) occurred in 2018. In contrast, captures of razor clam (Ensis siliqua) have increased by 3.6-fold since 2016. Predation risk for these taxa with contrasting burrowing capacities (1.7 ± 0.3cm, 0.4 ± 0.2cm, and 26.3 ± 0.1cm, respectively), was assessed using predation preference (N= 5 tanks; 5 individuals of each species) and no-choice experiments (N= 5 tanks; 15 individuals) in the absence and in the presence of sand. Results showed that in the absence of sand, the razor clam, was fully preyed in 24h, clams in 96h, and cockles reached 60% after 144h. Conversely, when sand was present, only 4% of razor clams were predated, while clams and cockles reached 60-100% in 120-144h. No-choice results featured similar patterns depending on substrate availability. Overall, clams and cockles appear to be greatly vulnerable to blue crab predation, whereas the razor clam may escape thanks to its deeper burrowing capacity.

: Poultry products in Zambia form an integral part of the human diet in many households, as they are cheap and easy to produce. The burden of poultry diseases has, however, remained a major challenge. Growing consumer demand for poultry products in Zambia has resulted in non-prudent antimicrobial use on farms, intending to prevent and treat poultry diseases for growth optimization and maximising profits. This cross-sectional study aimed to identify the different types of bacteria causing diseases in chickens in Lusaka and to detect the extended-spectrum-lactamase (ESBL)-encoding genes. We collected 215 samples from 91 diseased chickens at three post-mortem facilities and screened them for gram-negative bacteria. Of these samples, 103 tested positive for various clinically relevant Enterobacteriaceae, including Enterobacter (43/103, 41.7%), Escherichia coli (20/103, 19.4%), Salmonella (10/103, 9.7%), and Shigella (8/103, 7.8%). Other isolated bacteria included Yersinia, Morganella, Proteus, and Klebsiella, which accounted for 21.4%. E. coli, Enterobacter, Salmonella, and Shigella were subjected to antimicrobial susceptibility testing. The results revealed that E. coli, Enterobacter, and shigella were highly resistant to tetracycline, ampicillin, amoxicillin, and trimethoprim-sulfamethoxazole, while Salmonella showed complete susceptibility to all antibiotics. The observed resistance patterns correlated with antimicrobial usage estimated from sales data from a large-scale wholesale and retail company. Six (6/14, 42.9%) E. coli isolates tested positive for bla_CTX-M, whilst eight (8/14, 57.1%) Enterobacter samples tested positive for bla_TEM. Interestingly, four (4/6, 66.7%) of the E. coli isolates carrying bla_CTX-M-positive strains were also positive for bla_TEM. Sanger sequencing of the PCR products revealed that five (5/6,83.3%) of the abovementioned isolates possessed the bla_CTX-M-15 allele. The results suggest the presence of potentially pathogenic ESBL-producing Enterobacteriaceae in poultry, threatening public health. Keywords:. Enterobacteriaceae, bla_CTX-M, bla_TEM

The serendipitous discovery of antiparasitic drugs, such as quinine and artemisinin from plant origin reveals that the search for new chemical pharmacophores from medicinal plants is valuable. The present study sought to explore the antiplasmodial, antileishmanial, and antitrypanosomal activities of extracts from Lippia adoensis. Crude extracts of L. adoensis leaves and twigs, which were obtained by extraction using 70% ethanol in water, were assayed for antiplasmodial activity against P. falciparum 3D7 and Dd2 through the SYBR green I-based fluorescence assay; and for antileishmanial, antitrypanosomal, and cytotoxic effects on Leishmania donovani, Trypanosoma brucei brucei, and Vero cells, respectively, using the resazurin colorimetric assays. The phytochemical analysis of the extracts was performed using a liquid chromatography-mass spectrometry (LC-MS) feature-based detection and molecular networking flow on Global Natural Product Social (GNPS). As a result, the crude extracts from L. adoensis inhibited growth of P. falciparum (3D7 and Dd2) (IC50s; (3D7): 10.008 and 97.467 μg/mL; (Dd2): 29.48 and 26.96 μg/mL), L. Donovani (IC50s: 22.879-10.522 μg/mL), and T. brucei brucei (IC50s: 2.3085-55.06 μg/mL). The extracts were found to be non-cytotoxic to Vero cells, thus yielding median cytotoxic concentrations (CC50s) above 100 μg/mL. The LC-MS tandem molecular networking flow predicted that the extracts contain valsafungin A and bacillamidin on the first cluster, and fatty acids, ketone and aldehyde derivatives on the second cluster. Overall, the present study demonstrated the antiparasitic effects of L. adoensis extracts, thus justifying the use of this plant in the traditional treatment of fever and malaria conditions. Nevertheless, detailed metabolomic studies and antiparasitic mechanisms of action of the extracts are expected to unveil the potential antiparasitic hit compounds.

Spodopotera frugiperda is a worldwide invasive pest that has caused major economic damage. According to classical biological control theory, natural enemies that can control invasive pests come from the area of origin as the pests that have gone through coadaptation processes. However, the new association’s approach suggests that local natural enemies are equally capable to control invasive pests. Due to the lack of data on the association of S. frugiperda and local natural enemies, research was conducted through a rapid survey to study the population density of S. frugiperda in Yogyakarta, Indonesia and to study the diversity of parasitoid associated with S. frugiperda. The results showed the difference in S. frugiperda population and parasitism rate, with 15 parasitoid species associated with S. frugiperda. Four egg parasitoids, eight larval parasitoids, and three pupal parasitoids were found to be associated with S. frugiperda for three years after it was first discovered in Indonesia. A new association was found between S. frugiperda and three parasitoid species, consisting of two egg parasitoids (Platygasteridaesp.01 and Platygasteridaesp.02) and a larval parasitoid (Eupelmidaesp.01). A relatively new association was found between S. frugiperda and two larval parasitoids (Coccygidium sp. and Stenobracon sp.), and an existing association was found between seven species including an egg parasitoid Telenomus remus, three larval parasitoids (Cotesia sp., Microplitis sp., Campoletis sp.), and three pupal parasitoids (Brachymeria lasus, B. femorata, and Charops sp.). Telenomus remus is the most dominant parasitoid, with a higher abundance and parasitism rate. This study also reported the first findings of the association of S. frugiperda with the larval parasitoid Megaselia scalaris in Indonesia. The result suggests another method for biological control using the new association approach since local natural enemies can foster quick adaptation to invasive pests.

Seeds of Arazá and Acai, in addition to the byproduct of the Sacha inchi seed, after the extraction of the oil known as Sacha Inchi oil press-cake (SIOPC), are considered wastes and often discarded; however, they have valuable potential as sources of nutrients and bioactive compounds. We aimed to analyze, the bromatological, and chemical composition of these three byproducts of the fruit industry. Further cytotoxic, antioxidant, and antihypertensive activities of extracts ethanol: water were evaluated through in vitro tests. The Bromatological test showed that SIOPC had a low protein content (8.1%), while AS had a high fiber content (48.9%), however, were poorest in nutrients Ethanol: water extracts from all seeds exhibited diverse profiles of secondary metabolites, with significant concentrations of polyphenols (155.88 µg gallic acid/100 mg dry weight), flavonoids (411.69 µg quercetin/100 mg dry weight), and carotenoids in AS (43.66 µg ß-carotene/100 mg dry weight). AS and ACS extracts excelled in ABTS and DPPH radical scavenging assays, and AS showed high α-glucosidase inhibition, previously unreported in the literature. The results suggest that both seeds and extracts are sources of nutritional and bioactive metabolites with high antioxidant and hypoglycemic activity. that could be used in the food industry and the health sector. These by-products underline the importance of sustainable harnessing these sources of the native Amazonian Region

Candida (C.) infections represent a serious health risk for people affected by inflammatory bowel disease. An important fungal virulence factor is the capacity to form biofilms on the colonized surface of the host. This research study aimed to determine the effect of C. tropicalis and C. albicans co-infection on dextran sodium sulfate (DSS)-induced colitis in mice. Colitis severity was evaluated using histology and colonoscopy. Mice were mono-inoculated with C. albicans, C. tropicalis or co-challenged with both species. Mice were administered 3% DSS to induce acute colitis. Biofilm activity was assessed using (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl] 2H-tetrazoliumhydroxide (XTT) assay. Abundance of C. albicans in the colon tissues was assessed by immunohistochemistry. Co-challenged mice showed decreased colitis severity compared to mono-inoculated mice. The XTT assay demonstrated a marked decrease of C. albicans biofilm production in a C. albicans culture incubated with C. tropicalis supernatant. Immunohistochemical staining showed that C. albicans was more abundant in the mucosa of C. albicans mono-inoculated mice compared to the co-inoculated group. These data indicate an antagonistic microbial interaction between the two Candida species, where C. tropicalis may produce molecules capable of limiting the ability of C. albicans to adhere to the host intestinal surface leading to a reduction in biofilm formation.

Maintaining the high overall health of farmed animals is a central tenant of their well-being and care. Intense animal crowding in aquaculture promotes animal especially in the absence of straightforward methods for monitoring their health. Here, we used bacterial 16S ribosomal RNA gene sequencing to measure population dynamics occurring during P. salmonis outbreaks. Our findings revealed a complex bacterial community consisting of a previously undescribed core pathobiome. Notably, we detected Aliivibrio wodanis and Tenacibaculum dicentrarchi on the skin ulcers of salmon infected with P. salmonis, while Vibrio spp. were enriched on infected gills. The prevalence of these co-occurring networks indicates the influence that coinfection plays in P. salmonis pathogenicity.

Due to its application as an anti-cancer drug, pazopanib (1) has attracted the interest of many researchers and several studies on pazopanib synthesis have been reported over the years. This paper provides a novel route for synthesizing N,2,3-trimethyl-2H-indazol-6-amine (5) which is a crucial building block in the synthesis of pazopanib from 3-methyl-6-nitro-1H-indazole (6). By alternating between the reduction and two methylation steps compound 5 was obtained in comparable yield (55.0%) to what has been reported (54%). Noteworthy, the last step of N2-methylation also yielded N,N,2,3-tetramethyl-2H-indazol-6-amine (5’) as a new compound ever reported. Furthermore, the data presented in this paper can serve as instrumental for future research aimed at further refining the synthesis process of pazopanib.

This study aimed to assess the microbiological quality of set of fruits and vegetables sold on stalls in the streets and open markets of Dakar, the capital city of Senegal. Samples purchased in seven sites were analyzed to isolate E. coli, Salmonella spp. and Vibrio spp. Some primary samples with a positive microbiological culture were subjected to DNA extraction and subsequent metagenomic analysis. A total of 240 fresh fruit and vegetable samples were collected including lettuce (n=40), tomatoes (n=40), mango slices (n=40), onion slices (n=40), mint leaves (n=40), strawberries and grapes (respectively n=20). Of these samples, 50.83% (122/240) and 60.41% (145/240) was contaminated with Escherichia coli and Vibrio spp. respectively, while Salmonella was not isolated in any product. Levels of contamination with both E. coli (1.3 103 to 6 106 CFU/g) and Vibrio spp. (4.55 102 to 8.73 106 CFU/g) were significantly above the thresholds acceptable for human consumption. The most contaminated samples were lettuce with a prevalence of 98% (39/40) for E. coli and for Vibrio spp. followed by mint leaves with 100% (40/40) and 93% (37/40) of the samples containing for E. coli and Vibrio spp. respectively. Out of 46 samples sequenced, metagenomic analysis revealed high contamination rates for E. coli, Vibrio spp. and Salmonella spp. with 100%, 67.39% and 93.47% of prevalence respectively. On the other hand, the alpha diversity analysis shows a high bacterial diversity in lettuce and mint leaf samples while beta diversity analysis highlighted the presence of two major clusters. Our results stress the need of a surveillance system that extends this investigation to a national scale while increasing the number of sampling sites and products analyzed.

The increase in incidence of superficial fungal infections combined with the emergence of antifungal resistance represents both a global health challenge and a considerable economic burden. Recently, dermatophytes, the main culprit causing superficial fungal infections, have started to exhibit antifungal resistance. This can be observed in some of the most common species such as Trichophyton rubrum and Trichophyton mentagrophytes. Importantly, the new subspecies, known as Trichophyton indotineae, has been reported to show high resistance to terbinafine, a first-line treatment for dermatophyte infections. Compounding these issues is the realization that diagnosing the causative infectious agents requires using molecular analysis that goes beyond the conventional macroscopic and microscopic methods. These findings emphasize the importance of conducting antifungal susceptibility testing to select the appropriate antifungal necessary for successful treatment. Implementing these changes may improve clinical practices that combat resistant dermatophyte infections.

Despite recent improvements in the methods used to eradicate and minimize mosquito populations, they still pose severe threats to human health. This study aimed to investigate the Larvicidal potentials of silver nanoparticles of Ocimum gratissimum on wild Anopheles larvae from Lagos. The plant used in this study was collected from the southeastern part of Nigeria, Ibite-Olo in Ezeagu Local Government Area of Enugu State. Wild Anopheles mosquito larvae were collected from a field at Kosofe Local Government Area of Lagos State, Nigeria where resistant Anopheline populations have been reported in and around Lagos State were brought to the laboratory and kept under standard insectary conditions (25–29 °C;78–82% relative humidity). Aqueous extract of Ocimum gratissimum was made and subsequently into the silver nanoparticles. A bioassay test was performed according to WHO guidelines, with six different concentrations to assess the larvicidal activity and values obtained were subjected to log prohibit regression analysis to obtain LC₅₀ and values with 95% confidence limit. The results of the larvicidal activities of the silver nanoparticles of Ocimum gratissimum showed that the mortality rate was dose-dependent with the percentage mortality increasing at a direct proportional rate as with dose and time of exposure. Ocimum gratissimum gave LC₅₀ at 0.5 ppm and LC₉₅ at 2.5 ppm respectively. The biosynthesized silver nanoparticles augmented the larvicidal activity of the silver nanoparticles of Ocimum gratissimum on wild Anopheles larvae from Lagos. Therefore, this study showed that the silver nanoparticles of Ocimum gratissimum have the potency to be used against an insecticide-resistant mosquito vector and are a suitable candidate for the development of novel natural insecticides.

BACKGROUND: Qu-aroma is a critical quality indicator for medium-high temperature Daqu (MT-Daqu), significantly influencing the aroma profile of strong-aroma Baijiu (SAB). SAB is produced across various regions in China, each with unique environmental conditions and production techniques. These regional differences are likely to cause variations in the Qu-aroma of MT-Daqu. Despite this, comprehensive research on the variations in Qu-aroma across different regions has been lacking. This study aims to address this gap by utilizing Gas Chromatography-Olfactometry-Mass Spectrometry (GC-O-MS) and electronic nose (E-nose) technology, combined with sensory evaluation, to analyze the differences in Qu-aroma characteristics and volatile compound profiles of MT-Daqu from seven distinct production areas.RESULTS: Sensory analysis identified seven broad aroma categories and 25 specific aromas characterizing the Qu-aroma of MT-Daqu samples. Chen aroma and roasted aroma were notably more prevalent in MT-Daqu samples from Sichuan compared to those from other regions (P < 0.05), while the fermentation aroma was less prominent in Sichuan samples. In total, 123 volatile compounds were detected in MT-Daqu, with 46 classified as aroma-active compounds. Among these, 21 aroma-active compounds were identified as markers distinguishing Sichuan MT-Daqu from those produced in non-Sichuan regions, showing significant differences in their concentrations (P < 0.05). Additionally, six major aroma-active compounds-ethyl isovalerate, ethyl heptanoate, hexanal, 1-nonanal, (E)-2,4-decadienal, and 2,3-dimethyl-5-ethylpyrazine-were found to significantly contribute to the aroma differences between Sichuan and non-Sichuan MT-Daqu.CONCLUSION: This study delineates the distinct Qu-aroma characteristics of MT-Daqu from various SAB production areas and identifies the aroma-active compounds responsible for these differences. The insights gained from this research offer valuable guidance for optimizing flavor profiles in MT-Daqu production, potentially enhancing the overall quality and regional distinctiveness of the product.

Ixodes scapularis is a blood-feeding obligate ectoparasite responsible for transmitting the Lyme disease (LD) agent, Borrelia burgdorferi. During the feeding process, I. scapularis injects B. burgdorferi into the host along with its saliva, facilitating the transmission and colonization of the LD agent. Tick calreticulin (CRT) is one of the earliest tick saliva proteins identified and is currently utilized as a biomarker for tick bites. Our recent findings revealed elevated levels of CRT in the saliva proteome of B. burgdorferi-infected I. scapularis nymphs compared to uninfected ticks. Differential precipitation of proteins (DiffPOP) and LC-MS/MS analyses were used to identify the interactions between Ixs (I. scapularis) CRT and human plasma proteins and further exploring its potential role in shielding B. burgdorferi from complement killing. We observed that although yeast expressed recombinant (r) IxsCRT binds to the C1 complex (C1q, C1r, and C1s), the activator of complement via the classical cascade, it did not inhibit deposition of the membrane attack complex (MAC) via the classical pathway. Intriguingly, rIxsCRT binds intermediate complement proteins (C3, C5, and C9) and reduces MAC deposition through the lectin pathway. Despite inhibition of MAC deposition in the lectin pathway, rIxsCRT did not protect a serum sensitive B. burgdorferi strain (B314/pBBE22Luc) from complement-induced killing. As B. burgdorferi establishes a local dermal infection before disseminating to secondary organs, it is noteworthy that rIxsCRT promotes the replication of B. burgdorferi in culture. We hypothesize that rIxsCRT may contribute to the transmission and/or host colonization of B. burgdorferi by acting as a decoy activator of complement and by fostering B. burgdorferi replication at the transmission site.

Chronic kidney disease (CKD) is characterized by a continuous decline in kidney function, often persisting for years or even a lifetime. This study presents a new exploration into the extraction and application of insect chitosan (Tenebrio molitor) targeting hemodialysis (HD). Initially, the potential of insect-derived chitosan as a sustainable and effective alternative to traditional sources (i.e. crustaceans) is demonstrated. Efficient extraction methods were developed (totaling between 6 h and 12 h), producing chitosan. Firstly, chitin was efficiently isolated from T. molitor through deproteinization and demineralization, with yields of approximately 5% (w/w). Subsequently, chitosan was obtained from the extracted chitin resulting in yields of between 65.0 ± 0.8% (w/w) and 79.3 ± 0.8% (w/w). Characterization using Fourier Transform Infrared Spectroscopy (FTIR) confirmed structural similarities with commercial chitosan and degrees of deacetylation in the 73 - 75% range. The bioactive properties of chitosan obtained from T. molitor, including antimicrobial and antioxidant activities, were evaluated. All the microorganisms tested were inhibited, exhibiting minimum lethal concentrations between 2 and 8 mg/mL, confirming the antimicrobial activity of the extracted chitosan. In addition, chitosan showed antioxidant activity in the range of 60 to 65 µmol Trolox equivalent/g, suggesting its viability for various medical applications.This study also developed a chitosan-based membrane (CH - M) culminating in a proof-of-concept phase for hemodialysis. The permeation characteristics of CH-M for urea and albumin were studied in vitro to assess their suitability as HD membranes. Urea was permeable to values of over 70% and albumin was retained.

Paratuberculosis, or Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic granulomatous enteritis affecting both domestic and wild ruminants. The agent was also found in wild mammals such as wild boar (Sus scrofa), however, the role of wild mammals in the epidemiology of MAP is unclear. During the research period, 941 free-ranging wild boar (S. scrofa) legally hunted in two locations in the Centre-eastern region of Portugal were examined. Ninety-seven wild boar exhibited one or more gross lesions and were tested for the presence of Mycobacterium avium subsp. paratuberculosis using acid-fast staining, mycobacterial culture, poly-merase chain reaction (PCR), and histopathological examination. Forty-five animals (46.4%, 95% CI: 36.5-56.3%) were identified as infected, as indicated by positive results in culture and/or PCR. The findings revealed that the most significant risk factor was being a juvenile compared to year-lings and adults (OR = 10.2, 95% CI: 2.2-48.0). Based on our results, 37.9% (n = 11) of the infected animals were considered suitable for human consumption. Our findings offer novel insights into mycobacterial infections in wild boar populations in Portugal and suggest that wild boar could be a source of human infection if zoonotic potential is considered.

Metagenomic analysis of Aedes aegypti and Ae. albopictus mosquitoes from diverse geographical regions of India revealed presence of several insect viruses of human interest. Most abundant reads found in Ae. aegypti mosquitoes were Phasi-Charoen like virus (PCLV), Choristoneura fumiferana granulovirus (CfGV), Cell fusing agent virus (CFAV) and Wenzhou sobemo-like virus 4 (WSLV4), whereas WSLV4 and CfGV constituted the highest percentage in Ae. albopictus viromes. Other reads which were of low percentage included Hubei mosquito virus 2, Porcine astrovirus 4 and Wild Boar astrovirus. PCLV and CFAV, which were found to be abundant in Ae. aegypti viromes, were absent in Ae. albopictus viromes. Among the viromes analyzed, Ae. aegypti sampled from Pune showed highest percentage (79.82%) of viral reads while Ae. aegypti mosquitoes sampled from Dibrugarh showed lowest percentage (3.47%). Shamonda orthobunyavirus, African swine fever virus, Aroa virus and Ilheus virus, having potential to infect vertebrates including humans, were also detected in both mosquito species, albeit in low read numbers. Reads of gemykibivirus, avian retrovirus, bacteriophages, herpesviruses and viruses infecting protozoans, algae, etc. were also detected in the mosquitoes. A high percentage of reads in the Ae. albopictus mosquito samples remained unclassified and warrant further investigation. Data generated in the present work may not only lead to studies to explain the influence of these viruses on replication and transmission of viruses of clinical importance but also find application as biocontrol agents against pathogenic viruses.

The predatory mirids Macrolophus pygmaeus Ramber and Nesidiocoris tenuis Reuter (Heteroptera: Miridae) are used for the biological control of Tuta absoluta Meyrick (Lep.: Gelechiidae) and other pests in tomato greenhouses. The functional response of 1 (young) and 10 (old) days old adult females and males of M. pygmaeus and N. tenuis on eggs of T. absoluta, was determined on tomato at two temperatures (20° and 25°C) and LD 16:8 and found to be Type II. M. pygmaeus females exhibited higher predation efficiency, than males at both tested temperatures. Young M. pygmaeus females had a higher efficiency than old ones, whereas males had a low efficiency irrespective of age. The predation efficiency of N. tenuis was high (but lower than M. pygmaeus) in both young females and males, although old females had a higher efficiency than the respective males. Our results show that the two predatory species have different functional response characteristics to their prey depending mainly on sex and age which may affect their role as biological control agents.

Burkholderia are a group of Gram-negative bacteria that can cause a variety of diseases in at-risk populations. B. pseudomallei and B. mallei, the etiological agents of melioidosis and glanders, respectively, are the two clinically relevant members of the B. pseudomallei complex (Bpc). The development of vaccines against Bpc species has been accelerated in recent years, resulting in numerous promising subunits and glycoconjugate vaccines incorporating a variety of antigens. However, a second group of pathogenic Burkholderia species exists known as the Burkholderia cepacia complex (Bcc), a group of opportunistic bacteria which tend to affect individuals with weakened immunity or cystic fibrosis. To date there have been few attempts to develop vaccines to Bcc species. Therefore, the primary goal of this review is to provide a broad overview of the various subunit antigens that have been tested in Bpc species, their protective efficacy, study limitations, and known or suspected mechanisms of protection. Then, we assess the reviewed Bpc antigens for their amino acid sequence conservation to homologous proteins found in Bcc species. We propose that protective Bpc antigens with a high degree of Bpc-to-Bcc sequence conservation could serve as components of a pan-Burkholderia vaccine capable of protecting against both disease-causing groups.

Fermented food matrices, including beverages, can be defined as the result of the activity of complex microbial ecosystems where different microorganisms interact according to different biotic and abiotic factors. Certainly, in industrial production the technological processes aim to control the fermentation to place safe foods on the market. Therefore, if food safety is the essential prerogative, consumers are increasingly oriented towards a healthy and conscious diet driving the production and consequently the applied research towards natural processes. In this regard, the aim to guarantee safety, quality and diversify of products should be reached limiting or avoiding the addition of antimicrobials or synthetic additives using the biological approach. In this paper, following the recent re-evaluation of non-Saccharomyces yeasts (NSYs) has been re-viewed the bio-protectant and biocontrol activity with a particular focus on their antimicrobial power, probiotic features and promoting functional aspects. In this review, the authors would underline the contribution of NSYs in all food production chain and its role in the technological and fermentative features for their practical and useful use as biocontrol agent in food preparations.

Lactic acid (LA) is a significant byproduct in Baijiu fermentation, with both D(-)-lactic acid (D(-)-LA) and L-lactic acid (L(+)-LA) being present due to the diverse microbial metabolism involved. L(+)-LA, a key industrial raw material, is metabolizable by humans, unlike D(-)-LA. This study aims to isolate microorganisms from fermented grains that preferentially produce L(+)-LA and apply these strains to Baijiu fermentation processes. We analyzed 106 commercially available Baijiu samples, along with fermented grains, yellow water, and Baijiu samples from 10 Baijiu factories, confirming the coexistence of D(-)-LA and L(+)-LA in fermentation products. Notably, a strong correlation was found between L(+)-LA in fermented grains and L-ethyl lactate (L(+)-EL) in Baijiu. Microbial community analysis revealed that Lactobacillus and Bacillus were the dominant bacterial groups, with a succession of microbial communities throughout fermentation. Two strains, A6 and B12, identified as Bacillus licheniformis and Bacillus cereus, respectively, were isolated for their high selectivity in L(+)-LA production. Both strains demonstrated a high yield of L(+)-LA in fermentation tests. Incorporating these strains into both liquid- and solid-state fermentation systems markedly increased the proportion of L(+)-EL relative to total ethyl lactate in Baijiu. This study successfully isolated and applied L(+)-LA-producing strains to fermentation models, offering a practical strategy for increasing L(+)-LA content in Baijiu.

Essential oils oregano (Origanum vulgare), rosemary (Salvia rosmarinus), clove (Syzygium aromaticum), thyme (Thymus), cinnamon (Cinnamomum verum), basil (Ocimum basilicum), have antifungal properties. This study evaluated its ability to inhibit growth of isolated fungi and purified obtained from the rot of the banana peel, for the purpose of combating or inhibiting fungal growth in bananas (Musa paradisiaca). The methodology consists in preparing the dilutions to inoculate on PDA medium (Potato Dextrose Agar) amended with chloramphenicol to re-inoculate until obtaining the isolated and purified fungi of Trichoderma spp., Aspergillus spp., Penicillium spp. and Fusarium spp., which are characterized macroscopically and microscopically, subsequently, the growth is analyzed in vivo and the inhibition percentage in vitro. The results of the analysis in vivo report that the fungi in order of severity from greatest to least, was Penicillium spp., Trichoderma spp, Fusarium spp. y Aspergillus spp., while that the analysis in vivo reports that the rosemary and the basil does not inhibit fungal growth, while that the clove inhibit a 800 ppm, the cinnamon to 400 ppm and the oregano to 200 ppm to the four fungi. -2077601433541Citation: To be added by editorial staff during production. Academic Editor: Firstname Lastname Received: date Revised: date Accepted: date Published: date Copyright: © 2024 by the authors. Submitted for possible open access publication under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).00Citation: To be added by editorial staff during production. Academic Editor: Firstname Lastname Received: date Revised: date Accepted: date Published: date Copyright: © 2024 by the authors. Submitted for possible open access publication under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

This article discusses advances in molecular technologies for detecting foodborne pathogens, specifically Listeria monocytogenes and Salmonella. Although traditional detection methods are reliable, they are time- and labor-intensive. On the other hand, molecular methods include polymerase chain reaction (PCR), quantitative real-time PCR (qPCR), and next-generation sequencing (NGS), which enable rapid, specific, and sensitive detection of pathogens. The introduction of such procedures has significantly improved food safety testing and monitoring. Although challenges such as accessibility and cost remain, future developments in molecular diagnostics will lead to greater implementation in improving pathogen management throughout the food supply chain.

Butirosins are naturally occurring aminoglycoside (AG) antibiotics featuring a 4,5-disubstituted 2-deoxystreptamine (2-DOS) with a (2S)-4-amino-2-hydroxybutyrate (AHBA) side chain. This side chain has been shown to confer resistance against AG modifying enzymes, leading to ongoing studies on the butirosin biosynthetic pathway and the corresponding enzymes. Butirosin is produced by Niallia (formerly Bacillus) circulans and Bacillus vitellinus, with most research focused on the former strain. To date, no whole genome analysis has been performed on B. vitellinus. In this study, we sequenced the complete genome of B. vitellinus NBRC 13296 and performed a comparative analysis of different butirosin biosyntheric gene clusters (BGCs), including those from N. circulans. The complete genome of B. vitellinus NBRC 13296 comprises a 6,331,192 base circlular chromosome with a GC content of 52.68%. The annotation revealed the presence of 5,605 CDSs, 70 tRNA genes, 30 rRNA genes, and 3 ncRNA genes in NBRC 13296. The highest dDDH and ANI values between NBRC 13296 and the most closely related type strain, Paenibacillus chitinolyticus KCCM 41400, were 97.8% and 98.66%, respectively. Based on these genome-based comparative analyses, we propose reclassifying B. vitellinus NBRC 13296 as P. chitinolyticus. Genome mining revealed 18 gene clusters encoding the biosynthesis of diverse secondary metabolites in the genome of B. vitellinus NBRC 13296, indicating the enormous biosynthetic potential of this strain. The predicted structural diversity of secondary metabolites includes aminoglycoside, PKS, NRPS, PKS-NRPS hybrids, metallophores, phosphonates, terpenes, β-lactones, and RiPP peptides. We then comparatively characterized butorosin BGCs previously studied in several N. circulans strains. Additionally, comparative genome analysis revealed complete butirosin BGCs identified from P. chitinolyticus KCCM 41400, P. chitinolyticus NRRL B-23119, P. chitinolyticus NRRL B-23120, P. chitinolyticus B-14908, P. chitinolyticus YSY-3.1, P. chitinolyticus JMW06, Paenibacillus sp. GbtcB18, Paenibacillus sp. HGH0039, Paenibacillus sp. MZ04-78.2. Finally, we identified the core region consisting of BtrS, BtrN, BtrM, BtrL, BtrA, BtrB, BtrC, BtrD, BtrD, BtrE, BtrF, BtrG, BtrH, BtrI, BtrI, BtrJ, BtrK, BtrO, BtrP, and BtrV, followed by an upstream region organizing BtrQ, BtrW, BtrX, BtrY, and BtrZ in the same transcriptional direction and sequential genetic arrangement, and a downstream region organizing various proteins based on BtrT, BtrR2, BtrU, and BtrR1. Our study provided insights into the reclassification of B. vitellinus NBRC 13296 to P. chitinolyticus and suggested the need for continued studies on butirosin biosynthesis from an enzymatic perspective.

Lecanicillium lecanii KMZW-1, is recognized for its potential as a bio-control agent against agricultural and forestry pests, particularly due to its compatibility with integrated pest management (IPM) strategies. This study aimed to investigate the complete genome of L. lecanii KMZW-1 and assess its pathogenicity against Bactrocera dorsalis. Whole genome sequencing revealed a genome size of 47,239,278 bp, comprising 27 contigs, with a GC content of 51.16%. The genome completeness was assessed as 97.93% using BUSCO analysis, AntiSMASH analysis identified 29 gene clusters associated with secondary metabolite biosynthesis providing insights into the genetic basis of its pathogenic mechanisms and biocontrol potential. The pathogenicity of L. lecanii KMZW-1 was assessed using concentrations ranging from 1×10^4 to 1×10^11 spores/mL. Higher-concentrations (1×10^10 to 1×10^11 spores/mL), resulted in significantly increased cumulative mortality rates of B. dorsalis adults compared to the control group. Notably, the pathogenicity was higher in male adults than in females. Probit regression analysis yielded LogLC50 values as 5.662 for female and 4.486 for male B. dorsalis adults. In summary, L. lecanii KMZW-1 exhibits strong insecticidal activity against B. dorsalis and shows potential as a biocontrol agent with IPM strategies. These findings provide robust genomic evidence supporting the use of L. lecanii KMZW-1in managing against B. dorsalis populations.

Pigeonpea (Cajanus cajan) is one of the important grain legume crops cultivated in the semi-arid tropics, playing a crucial role in the economic well-being of subsistence farmers. India is the major producer of pigeonpea, accounting for over 75 per cent of the world's production. Sterility mosaic disease (SMD), caused by Pigeonpea sterility mosaic virus (PPSMV) and transmitted by the eriophyid mite (Aceria cajani), is a major constraint to pigeonpea cultivation in the Indian subcontinent, leading to potential yield losses of up to 100 per cent. The recent characterization of another Emaravirus associated with SMD has further complicated the etiology of this challenging viral disease. This review focuses on critical areas, including the current status of the disease, transmission and host-range, rapid phenotyping techniques, as well as available disease management strategies. The review concludes with insights into the future prospects, offering an overview and direction for further research and management strategies.

The pressing threat of global warming poses significant risks to plant survival, adversely affecting their growth and agricultural productivity. It is crucial to understand how plants react to heat stress at the molecular level to develop crops that can withstand higher temperatures. In our study, we investigated the proteome dynamics of *Arabidopsis thaliana* seedlings under moderate heat stress conditions (30°C) using a combination of 15N-stable isotope labeling and the ProteinTurnover algorithm. This approach allowed us to closely examine the proteomic changes across different cellular compartments. Our findings revealed significant changes in the turnover rates of 571 proteins, with an average increase of 1.4-fold, indicating enhanced protein dynamics under heat stress. Notably, root soluble proteins showed more subtle changes, suggesting tissue-specific responses. We also detected significant turnover variations in proteins involved in redox signaling, stress response, and metabolism, highlighting the complex nature of these adaptive mechanisms. In contrast, proteins related to carbohydrate metabolism and mitochondrial ATP synthesis showed little change, underscoring their stability. This detailed analysis illuminates the proteomic adaptations of *Arabidopsis* seedlings to moderate heat stress, enhancing our understanding of plant thermal resilience and supporting efforts to develop thermotolerant crops.

Understanding mechanisms of ageing remains a complex challenge for biogerontologists, but recent adaptations of evolutionary ageing theories offer a compelling lens in which to view both age-related molecular and physiological deterioration. Ageing is commonly associated with a progressive loss of biochemical processes and degeneration of molecular material, however, the mechanisms of diminishing function are not simply a result of aberrant expression of these pathways. Natural selection pressures are at their highest in youthful periods to select genes advantageous to maximising reproductive capacity. After sexual maturation, selective pressure diminishes, subjecting individuals to maladaptive pleiotropic gene function once beneficial for developmental growth, but pathogenic later in life. Due to this selective ‘shadowing’ in ageing, mechanisms to counter hyperfunctional genes are unlikely to evolve within a given species. These genetic ‘run-on’ programmes are not dysfunctional per se and, in fact, remain highly robust. Thus, interventions capable of optimising gene function during the ageing process represent as attractive therapeutic modalities. Caenorhabditis elegans offers a unique model to explore high-throughput screening of interventions to combat hyperfunctional pathways involved in ageing. However, the majority of research to date employ whole-life vs late-life pathway manipulation of developmental/growth-related systems to claim ageing attenuation, despite significant impairments in early-life fitness and discordance with healthspan. Here, we discuss data supporting hyperfunctional changes at a global molecular and functional level in C. elegans, and how classical lifespan-extension mutants alter these dynamics. We later discuss the relevance of such mutant models for exploring mechanisms of ageing, highlighting that post-reproductive gene manipulation represents a far more translatable approach for C. elegans research not routed within pleiotropic constraints. More so, lifespan has become a routine readout for quantifying ‘ageing’, however, we make the argument that a compression of gerospan vs lifespan extension is a more meaningful goal for anti-ageing regimes with current knowledge. Similarly, mechanisms governing lifespan are distinct from those regulating health, and slowing senescent pathology accumulation appears more applicable for the abstraction to human ageing.

Individuals with BPD grapple with unstable relationships, marked by extreme emotional swings and interpersonal conflicts. Emotional instability, a distorted self-image, and impulsivity further complicate matters, often leading to self-harm and suicide attempts as desperate coping mechanisms. Genetics and adverse childhood experiences contribute to BPD's development, shaping brain function and emotional regulation. Diagnosis involves meticulous assessments, paving the way for personalized treatments like dialectical behavior therapy and, in severe cases, hospitalization. Early intervention, professional support, and societal understanding are pivotal for positive outcomes, and challenging mental health stigma.

The crayfish plague is a devastating disease of European freshwater crayfish and is caused by the oomycete Aphanomyces astaci (Ap. astaci), which was introduced to Europe around 1860. All European species of freshwater crayfish are susceptible to the disease, including the white-clawed crayfish Austropotamobius pallipes. The aquatic mould is primarily spread by North American crayfish species and can also disperse rapidly through contaminated wet gear moved between water bodies. This spread, coupled with competition from non-indigenous crayfish, has drastically reduced and fragmented native crayfish populations across Europe. Remarkably, the island of Ireland remained free from the crayfish plague pathogen for over 100 years, providing a refuge for A. pallipes. However, this changed in 1987 when a mass mortality event was linked to the pathogen marking the pathogen's introduction to the region. Fortunately, the plague did not establish and was not detected again in Ireland until 2015 when a molecular analysis linked a mass mortality event in the Erne catchment to Ap. astaci. Since then, the pathogen has appeared across the island. Between 2015 and 2023, Ap. astaci was detected in 18 Irish water catchments, revealing multiple genotypes. Intriguingly, the pathogen in Ireland exists without its natural host species. The uneven distribution of various genetic lineages strongly suggests human-mediated transport of zoospores via contaminated water equipment as a primary cause of spread. This review details the timeline of these events, Ap. astaci’s introductions into Ireland, its rapid spread, the genotypes documented, the issue of non-indigenous crayfish species in Ireland and management efforts.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts , but their overgrowth can lead to infection in the urogenital tract and at distal sites. Treatment of Ureaplasma spp. infections is difficult. The lack of a cell wall confers resistance to all β-lactam and glycopeptide antibiotics whereas the lack of de novo synthesis of folic acid makes them resistant to sulfonamides and diaminopyrimidines. Thus, there is a need to search for new drugs effective against these bacteria. In the presented studies, the effect of N,N-diethyldithiocarbamate (DDC) at a concentration of 2 µg/ml, 20 µg/ml and 200 µg/ml on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested. Our results indicated that all DDC concentrations used showed both bacteriostatic and bactericidal activity against both tested strains In brief, for the first time we showed the bacteriostatic and bactericidal activity of DDC against Ureplasma urealyticum and Ureaplasma parvum. Based on literature data, we conclude that this DDC activity is related to the inhibition of urease activity, which is an indispensable enzyme for these bacteria. Further research is needed and the results obtained in the present study should be verified in in vivo models of infection.

O26 enteropathogenic Escherichia coli (EPEC), possesses a unique pseudocapsule composed of polysaccharides identical to the O antigen in their lipopolysaccharide (LPS) membrane. Therefore, the impact of O26 polysaccharides on the immune evasion mechanisms of capsulated O26 EPEC was investigated. Our findings reveal that the concentration of GlcNAc, an immunodominant unit of O26 polysaccharides capable of activating the MBL (Mannose Biding Lecting) pathway of the complement system, was found to be lower in capsulated EPEC O26 compared to naked shiga toxin-producing E. coli (STEC) O26 strain. It was noted that, unlike the naked STEC strain, the capsulated EPEC O26 strain was protected against lysis by the alternative pathway of the com-plement system by inhibiting C3b adhesion to its bacterial surface. Capsulated EPEC O26 strains were also able to avoid phagocytosis by macrophages. However, the immune evasion provided by the capsule was overcome in the presence of anti-O26 polysaccharide antibodies. Additionally, these antibodies were able to recognize O26 EPEC in the presence of biofilm and inhibit their adhesion to human epithelial cells. Overall, the results indicate that the O26 polysaccharides can generate an effective humoral immune response, making them promising antigens for the de-velopment of a vaccine against capsulated O26 E. coli

Thanks to several Vitis vinifera backcrosses with an initial V. vinifera L. × V. rotundifolia (previously Muscadinia rotundifolia) interspecific cross, the MrRUN1/MrRPV1 locus (resistance to downy and powdery mildews) was introgressed in genotypes phenotypically close to V. vinifera varieties. To check the consequences of introgressing parts of the V. rotundifolia genome on gene expression during fruit development, we conducted a comparative RNA-seq study on single berries from different V. vinifera cultivars and V. vinifera × V. rotundifolia hybrids, including ‘G5’ and two derivative microvine lines, ‘MV102’ (resistant) and ‘MV32’ (susceptible) segregating for the MrRUN1/RPV1 locus. RNA-Seq profiles were analyzed on a comprehensive set of single berries from the end of the herbaceous plateau to the ripe stage. Pair-end reads were aligned both on V. vinifera PN40024.v4 reference genome, V. rotundifolia cv ‘Trayshed’ and cv ‘Carlos’, and to the few resistance genes from the original V. rotundifolia cv ‘52’ parent available at NCBI. Weighted Gene Co-expression Network Analysis (WGCNA) led to classifying the differentially expressed genes into 15 modules either preferentially correlated with resistance or berry phenology and composition. Resistance positively correlated transcripts predominantly mapped on the 4-5 Mb distal region of V. rotundifolia chromosome 12 beginning with the MrRUN1/MrRPV1 locus, while the negatively correlated ones mapped on the orthologous V. vinifera region, showing this large extremity of LG12 remained recalcitrant to internal recombination during the successive backcrosses. Some constitutively expressed V. rotundifolia genes were also observed at lower density outside this region. Genes overexpressed in developing berries from resistant accessions, either introgressed from V. rotundifolia, or triggered by these in the vinifera genome, spanned various functional groups, encompassing calcium signal transduction, hormone signaling, transcription factors, plant–pathogen-associated interactions, disease resistance proteins, ROS and phenylpropanoid biosynthesis. This transcriptomic insight provides a foundation for understanding the disease resistance inherent in these hybrid cultivars and suggests a constitutive expression of NIR NBS LRR triggering calcium signaling. Moreover, these results illustrate the magnitude of transcriptomic changes caused by the introgressed V. rotundifolia background in backcrossed hybrids, on a large number of functions largely exceeding the ones constitutively expressed in single resistant gene transformants.

Ethanolic extracts of propolis collected from Apis mellifera hives have been shown to reduce Nosema ceranae infection in honey bee workers. This study aimed to determine the propolis concentration that optimizes the control of N. ceranae infection in honey bee workers. Newly emerged worker bees were individually fed 2 μL of 50% sucrose solution (w/v) containing 105 N. ceranae spores per bee, and treated with 0%, 25%, 50%, and 75% ethanolic extracted propolis in 50% sucrose solution (v/v). The treated bees were maintained in cages at 34 ± 2˚C and 55 ± 5% RH throughout the study. The number of dead bees was counted and removed from the cages daily for 30 days to generate survival curves. The number of N. ceranae spores per bee (infectivity) was quantified using a hemocytometer, and the number of infected bees per hundred bees (infection rate) was determined. The survival rates of N. ceranae infected bees fed 25%, 50%, and 75% propolis (NP1, NP2, NP3, respectively) were significantly higher than those of infected bees not fed propolis (NO). Furthermore, the infectivity and infection rates of NP2 and NP3 groups were significantly lower than those of the NO group. The infectivity and infection rates of the NP1 group were not different than those of the NO group. Our data suggest that 50% and 75% propolis extracts are appropriate concentrations for treating N. ceranae infection in honey bees. Ethanolic extracted propolis can be an alternative treatment for N. ceranae infection in Western honey bees.

This study developed Makgeolli (Korean traditional alcoholic beverage) by adding raw 20% Codium fragile (Cf), a green seaweed, to a mixed base consisting of rice and oats (1:1 ratio) (COM) and analyzed its quality characteristics and antioxidant activity. The rice based Makgeolli supplemented with raw 20% Cf was designated as control (CRM). The physicochemical (pH, acidity), microbiological (yeast, lactic acid bacteria), antioxidant (DPPH, ABTS), and sensory properties (7-henodonic scale) of CRM and COM were investigated during 7 days of fermentation. The pH/acidity levels of CRM and COM reached 3.66/0.68 and 3.69/0.95 after 7 days of fermentation, respectively. Compared to CRM, COM had lower alcohol and sugar contents (P < 0.05) (CRM: 14.5%/9.8 Brix, COM: 11.2%/7.4 Brix). However, COM contained more yeast and lactic acid bacteria (P < 0.05) (CRM: 1.30/4.28 log, COM: 6.04/6.09 log) compared to CRM. The antioxidant activity of DPPH/ABTS (P < 0.05) in COM was approximately 1.5 times that of CRM (CRM: 40.00/51.57%, COM: 56.76/70.91%). In addition, COM had an excellent sensory preference score (P < 0.05) due to its overall fresh flavor and savory taste (CRM: 4.93, COM: 6.06). Therefore, COM was shown to be Makgeolli with enhanced antioxidant function, with yeast and lactic acid bacteria, and improved flavor, suggesting the possibility of commercializing this Makgeolli.

Megaselia scalaris and Senotainia tricuspis are two parasitoid flies of the honeybee Apis mellifera and are known to infest apiaries of different European and Mediterranean countries. However, their prevalence is still little known and no systematic control of their presence and countermeasures are currently adopted. To achieve a better knowledge about their distribution in apiaries, quantitative PCR (qPCR) based methods for their rapid detection directly in hive matrices were developed in this study. These were applied to samples of adult honeybees and hive debris collected in the summer season in Central Italy and Kosovo. In total 16 samples per type per region were analyzed. Both flies were detected in the two geographical contexts but more frequently in Central Italy. The positive apiaries in Central Italy were re-sampled one year later by capturing viable honeybees and isolating emerging flies after differentiation into adults. These were genotyped by sequencing regions of the mitochondrial cytochrome oxidase subunit I gene COI for M. scalaris and cytochrome B gene cytB for S. tricuspis amplified with specific primers designed in this study. Different genotypes of M. scalaris were detected and these were related to biotypes from distant nations. Some fly isolates were shown to harbor DNA or RNA of honeybee microbial pathogens. Results suggested to implement efficient control against these parasites in apiaries to limit their detrimental effects. The qPCR tests that have been developed can facilitate larger screening studies and early detection.

Finding, safe and reliable alternatives to fungicides is one of the most actual challenges in agriculture. In this regard, our experiment investigated the effectiveness of different elicitors, botanical extracts and essential oils to control grey mould (Botrytis cinerea) and powdery mildew (Podosphaera aphanis) on strawberry plants. Furthermore, the defence mechanisms activated in plants after treatments were investigated. The trial was conducted in field conditions under plastic tunnel with strawberry plants ‘Elsanta’. In the first experiment, a group of strawberry plants was treated before flowering with elicitors [acibenzolar-S-Methyl – (BTH), chitosan], botanical extracts (seaweed extract, alfalfa hydrolysate) and essential oils (thyme and juniper) to evaluate their ability to reduce the incidence of grey mould infection on flowers. In the second experiment, another group of plants was treated before and after experimental inoculation with P. aphanis. Results indicated that the incidence of flower infected by B. cinerea was significantly reduced by thyme and juniper essential oils, showing values that were not significantly different from the commercial fungicide penconazole (positive control). As a consequence, final yield of essential oil-treated plants was +27% higher as compared to control. No significant differences emerged for other tested products against grey mould. However, gene expression analysis showed an up-regulation of FaEDS1, FaLOX and PR gene expression (FaPR1, FaPR5, FaPR10) in leaves treated with BTH. The other tested natural substances also elicited defence-related genes, however at a lower level than BTH. In the experiment, all treatments applied prior inoculation significantly reduced the incidence and severity of powdery mildew as compared to control. At 28 days after inoculation, chitosan and thyme essential oil applications performed similarly to their positive controls (BTH and penconazole, respectively), showing a significant reduction of disease incidence (by -84 and -92%) as compared to control. Post-inoculum application of essential oils showed similar effective action to fungicide (penconazole) in reducing incidence and severity of powdery mildew. These results indicate that the tested substances could be used as alternatives to fungicides for the control of grey mould and powdery mildew in strawberry, therefore representing a valuable tool for the control of these fungal diseases under the framework of the sustainable agriculture.

Euler sums are alternating (or level two) extension of multiple zeta values (MZVs). Kaneko and Tsumura initiated the study of multiple T-values (MTVs), another level two generalization, by restricting the summation indices in the definition of MZVs to a fixed parity pattern. In this paper, we shall study finite MTVs and their alternating versions which are level two and level four variations of finite MZVs, respectively. We conjecture that all finite MZVs are in the Q-span of finite MTVs which in turn apparently lie in the span of finite Euler sums, and the inclusions are both proper. We shall first provide some structural results for Euler sums of small weights, guided by the author's previous conjecture that the finite Euler sum space of weight w is isomorphic to a quotient Euler sum space of weight w. Then, by utilizing some well-known properties of the classical alternating MTVs, we shall derive a few important Q-linear relations among the finite alternating MTVs, including the reversal, linear shuffle and sum relations. We then compute the upper bound for the dimension of the Q-span of weight w finite (alternating) MTVs for w<9, both rigorously using the newly discovered relations and numerically aided by computers.

This study is based on the clamor of fish farmers for a cheaper fish feed alternative to current commercial feeds. At present, 70-80% of aquaculture costs come from feed cost, keeping farmers' income below their expectation. This study aimed to find alternative feeds or supplementary feeds that can perform similarly to the existing commercial feeds thereby reducing farmer expense and cutting down their cost of production and increasing their profitability. To do that, various indigenous raw materials were selected based on an earlier study which could result in a better feed alternative. The formulations for the study included four grow-out treatments mainly: control (commercial feed), taro+control (powdered Colocasia esculenta+commercial fish feed), bloodmeal (formulated fish feed based on bloodmeal), banana stem+control (chopped banana stem+commercial fish feed). A total of 12,000 fingerlings initially weighing 16 g (TL=10 cm) individually were assigned randomly to the four treatments (3000 in each cage measuring 6 x 6 m). Week zero was used for acclimation period, and fingerlings were fed with commercial fish feed. Then the first week used the treatment diets which were adjusted based on average body weight and sampled weekly (ABW=23g; TL=11 cm). The result of the study first showed significant differences between treatments on their fourth week in terms of the diet treatments (MS=0.103, df=3, F=4.12, P=0.0076) with the control and the taro+control showing no difference in terms of weight and length (TC vs CF; P=0.4524). But during the fifth week, the bloodmeal treatment showed increased weight and length for fish sampled (MS=0.109, df=3, F=4.10, P=0.0076) and provided similar weight and length as those from the taro+control and the control (TC vs CF; P=0.3721 and BM vs CF; P=0.0587). During the succeeding weeks, there were no significant differences between feed treatments from 6th weeks onwards, whether with weight [MS=0.130, df=3, F=1.14, P=0.333] or total length of the treatments [MS=0.008, df=3, F=0.40, P=0.75]. The FCR ratio showed taro (1.81) as lowest, followed by blood meal (2.09), and control (2.24) and then banana stem (2.84). In terms of weight gain, fish from the bloodmeal treatment gained the highest (236.97 g), followed by control (223.33 g), then taro (217.67 g) and banana stem (196.99 g). As a result, we find that the tested treatments such as bloodmeal formulated feed, taro+commercial feed, and banana stem+commercial feed can serve as an alternative to purely commercial fish feeds. Cost-benefit analysis showed that a better gross profit margin of 42% and cost-benefit ratio of 1.37 was observed in taro+commercial feed diet.

We studied yeast hand carriage of 260 students from medicine (78), nursing (89), and medical laboratory scientist (93). We isolated yeasts in 27 students (10.4%), with no differences between careers or gender. A significant lower prevalence of carriage was shown in the clinical cycle (2.7%) compared to the basic cycle (13.5%) (p=0.022) and the preclinical cycle (13.5%) (p=0.014).Increased handwashing frequency and alcohol gel use decreased yeast carriage. Students who applied moisturizing hand cream two or more times a day had a lower frequency of yeast carriage (3.4%) compared to those who did not use it or used it only once a day (16.5%), showing a significant difference (p=0.016). The most prevalent species was C. parapsilosis sensu stricto (81.5%), followed by Meyerozyma guilliermondii (C. guillermondii) (7.4%), Trichosporon mucoides (7.4%), and R. mucilagenosa (3.7%). And one case showed mixed carriage of C. parapsilosis and C. albicans. All strains were sensitive to voriconazole, caspofungin, and anidulafungin. This study shows the yeast carriage on the hands of health students, especially of C. parapsilosis, and its reduction with the frequent use of infection control measures and moisturizing hand cream.

Indole-3-acetic acid (IAA), as an important regulator of potato growth, seriously affects the growth and yield of potato. Although many studies have reported that IAA-producing Bacillus can promote plant growth, little research has been done on its synthesis pathway and the molecular mechanisms. In this study, IAA-producing strain WL35 was identified as Fitibacillus barbaricus, and its yield was 48.79 mg·L-1. The results of the pot experiments showed that WL35 significantly increased plant height, stem thickness, chlorophyll content and number of leaves of potato plants by 31.68%, 30.03%, 32.93% and 36.59%, respectively. In addition, in the field experiments, WL35-treated plants increased commercial potatoes yield by 16.45%, vitamin C content by 16.35%, protein content by 75%, starch content by 6.60%, nitrogen, phosphorus and potassium accumulation by 9.98%, 12.70% and 26.76%, respectively. Meanwhile, the synthetic pathway of WL35 was found to be dominated by the tryptophan-dependent pathway, the IAM, TAM and IPA pathways worked together, and the pathways that played a role at different times were different. Furthermore, RNA-seq analysis showed that there were a total of 2875 DEGs regulated in the samples treated with WL35 seed dressing compared with the CK, of which 1458 genes were up-regulated and 1417 genes were down-regulated. Potato roots express differential genes enriched in processes such as carbohydrate metabolism processes and cellular polysaccharide metabolism, which regulate potato plant growth and development. The above results provide a theoretical basis for the further exploration of the synthesis pathway of IAA and its growth-promoting mechanism on potato.

Amphibians are widely known as a prolific source of bioactive metabolites. In this work we isolated and characterized compounds with antiparasitic activity from the oocytes of the toad Rhinella alata collected in Panama. Bio-guided isolation and structural elucidation was carried out using chromatographic and spectroscopic techniques. The organic crude extract was subjected to solid phase extraction followed by HPLC purification of the fraction with in vitro activity against Trypanosoma cruzi trypomastigotes. Seven steroids (1-7) of the bufadienolide family were isolated and their structures determined using NMR and MS analyses; of these 19-formyl-dyscinobufotalin (3) is reported as a new natural product. Compounds 1, 3-7, resulted with a good anti-trypanosomal activity profile, among these 16β-hydroxyl-hellebrigenin (1) and bufalin (7) showed significant selectivity for T. cruzi. Furthermore, molecular docking analysis showed compounds 1, 3 and 7 interact through H-bonds with the amino acid residues GLN-19, ASP-158, HIS-159 and TRP-177 from cruzipain, at the catalytic site. Given the lack of therapeutic options to currently treat the American trypanosomiasis, this work can serve as the basis for further studies that aim the development of bufadienolides or their derivatives as drugs against Chagas disease.

In the last decade, the application of non-Saccharomyces yeasts in oenology as a natural tool to obtain wine diversification and higher quality has aroused great interest. In this work, three M. pulcherrima strains, isolated from vineyard, were characterized through the evaluation of their main technological properties, antimicrobial activity and specific enzymatic activities (β-glucosidase, β-lyase, polygalacturonase and protease). The M. pulcherrima strains did not produce any inhibition against S. cerevisiae, while they were able to exert an antimicrobial action against some unwanted bacteria and yeasts frequently present in grape must and potentially causing alteration of wines. After this preliminary screening, M. pulcherrima AS3C1 has been selected to be used in the winemaking of red grape Vitis vinifera cv. Aglianico on a pilot scale. The effect of sequential inoculation of M. pulcherrima AS3C1 with a commercial strain of Saccharomyces cerevisiae was verified using for comparison a single inoculum with S. cerevisiae and a spontaneous fermentation. Our results showed a higher concentration of anthocyanins and catechins in wines obtained by sequential inoculation of M. pulcherrima AS3C1 and S. cerevisiae. On the basis of the data obtained, M. pulcherrima AS3C1 possesses an enzymatic profile and some oenological properties that could contribute positively to the definition of the chemical composition of wines, suggesting its possible use for red winemaking processes.

Water is essential for all life on earth. It is a major component that makes up living organisms and plays a vital role in multiple biological processes. It provides a medium for chemical and enzymatic reactions in the cell and is a major player for osmoregulation and maintenance of cell turgidity. Despite this, many organisms, called anhydrobiotes, are capable of surviving under extremely dehydrated conditions. Less is known about how anhydrobiotes adapt and survive under the desiccation stress. Studies have shown that morphological and physiological changes occur in anhydrobiotes in response to desiccation stress. Certain disaccharides and proteins, including heat shock proteins, intrinsically disordered proteins, and hydrophilins, play important roles in the desiccation tolerance of the anhydrobiotes. In this review, we summarize the recent findings of the desiccation tolerance in the budding yeasts Saccharomyces cerevisiae. We also propose that the yeast under desiccation could be used as a model to study neurodegenerative disorders.

Many contaminant yeast strains able to survive inside fuel ethanol industrial vats show detrimental cell surface phenotypes, such as filamentation, invasive growth, flocculation, biofilm formation and excessive foam production. Previous studies have linked some of these phenotypes to the expression of FLO genes, and the presence of gene length polymorphisms causing the expansion of FLO gene size appears to result in stronger flocculation and biofilm formation phenotypes. We have performed here a molecular analysis of FLO1 and FLO11 gene polymorphisms present in contaminant strains of S. cerevisae from Brazilian fuel ethanol distilleries showing strong foaming phenotypes during fermentation. The size variability of these genes was correlated with cellular hydrophobicity, flocculation and highly foaming phenotypes in these yeast strains. Our results also show that deleting the major activator of FLO genes (the FLO8 gene) from the genome of a contaminant and highly foaming industrial strain avoids problematic foam formation, flocculation, invasive growth and biofilm production by the engineered (flo8∆::BleR / flo8Δ::kanMX) yeast strain. Thus, the characterization of highly foaming yeasts and the influence of FLO8 in this phenotype opens new perspectives for yeast strain engineering and optimization in the sugarcane fuel-ethanol industry.

Beer is a fermented beverage with a history as old as human civilization and its productive process has been spread all around the world becoming unique in every country and iconic of entire populations. Ales and lagers are by far the most common beers; however, the combination of raw materials, manufacture techniques and aroma profiles are almost infinite, so it is not surprising to notice that there is a large amount of different beer styles, each of them with unique characteristics. Nowadays, diversification is becoming increasingly important in the brewing market and the brewers are continuously interested in improving and extending the already wide range of products, especially in craft brewery. One of the major components that can have a deep impact on the final product is yeast, since it is able to convert carbohydrates in wort, especially maltose and maltotriose, into ethanol, carbon dioxide and other minor aroma-active compounds. Saccharomyces cerevisiae (top‐fermenting yeasts used to produce ales) and Saccharomyces pastorianus (cryotolerant bottom‐fermenting hybrids between S. cerevisiae and Saccharomyces eubayanus responsible for the fermentation of lagers) are most used in breweries. However, an increasing number of different yeast starter cultures are commercially available, to improve the production efficiency also at relative low temperatures and to obtain desirable and diversified aroma profiles avoiding undesired compounds. Four main genetic engineering-free trends are becoming popular in craft brewing yeast development: 1) the research for novel reservoirs as source of new performant S. cerevisiae yeasts; 2) the creation of synthetic hybrids between S. cerevisiae and Saccharomyces non-cerevisiae in order to mimic lager yeasts by expanding their genetic background; 3) the exploitation of evolutionary engineering approaches; 4) the usage of non-Saccharomyces yeasts either in co-coculture or in sequential fermentation with S. cerevisiae. In the present work we summarized pro and contra of these approaches and provided an overview on the most recent advances on how brewing yeast genome evolved and domestication took place. Finally, we delineated how the correlations maps between genotypes and relevant brewing phenotypes can assist and further improve the search for novel craft beer starter yeasts.

Cytokinesis, the process by which the cytoplasm divides to generate two daughter cells after mitosis, is a crucial stage of the cell cycle. Successful cytokinesis must be coordinated with chromosome segregation and requires the fine orchestration of several processes, such as constriction of the actomyosin ring, membrane reorganisation, and, in fungi, cell wall deposition. In Saccharomyces cerevisiae, commonly known as budding yeast, septins play a pivotal role in the control of cytokinesis by assisting the assembly of the cytokinetic machinery at the division site and controlling its activity. Yeast septins form a collar at the division site that undergoes major dynamic transitions during the cell cycle. This review will discuss the functions of septins in yeast cytokinesis, their regulation and the implications of their dynamic remodelling for cell division.

Glucosinolates, the main secondary metabolites accumulated in cruciferous flora, have a major impact on fortifying plant immunity against diverse pathogens. Although Botrytis cinerea exhibits varying sensitivity to these compounds, current research has yet to fully understand the intricate mechanisms governing its response to glucosinolates. Different species of the genus Botrytis were exposed to glucosinolate-breakdown products revealing that B. fabae, B. deweyae and B. convoluta, species lacking the mfsG transporter gene (Bcin06g00026), were more sensitive to glucosin-olate-breakdown products than Botrytis species containing that gene such as B. cinerea, B. pseudo-cinerea and B. byssoidea. This finding was further corroborated by the inability of species lacking the mfsG gene to infect plants with a high concentration of glucosinolate breakdown products. These results challenge established correlations, revealing varying aggressiveness on different plant substrates. Expression analysis highlighted the gene's induction in the presence of isothiocyanate, and bioinformatic investigation identified homologous genes in other Botrytis species. Our study underscored the importance of advanced biotechnology to help understand these proteins and thus offer innovative solutions for agriculture

Leishmania infantum is an hemopathogen of importance for the health of domestic dogs (Canis lupus familiaris), causing canine leishmaniasis (CanL), as well as it is the etiological agent of human visceral leishmaniasis (HVL). This parasite was not reported in southern Brazil until the early 2000s, but CanL and HVL were increasingly reported in the last 15 years, mainly in cities bordering Argentina. The present study aimed to detect L. infantum in urban dogs and to determine the main clinical manifestations in the infected animals from Uruguaiana, a city with a high incidence of CanL. Fifty-one dogs suspected of having CanL in the urban perimeter of the city were clinically examined by veterinarians and investigated for the occurrence of L. infantum with two immunoassays (rapid chromatography test and ELISA) and real-time PCR. Clinical signs were compared in positive and negative L. infantum animals. A total of 31 dogs (60.8%) were infected with L. infantum. The main clinical manifestations associated to CanL dogs were onychogryphosis and peeling (p&lt;0.05). L. infantum was frequently detected in urban dogs from Uruguaiana, highlighting the health concerning situation in this city. The occurrence of some clinical signs (onychogryphosis/peeling) could help to detect more frequently CanL in the canine population.

This paper aimed to evaluate the growth, gas exchange and Chl a fluorescence in ipê-rosa seedlings, subjected to levels of 100, 75, 50 and 25% of water replacement (LWR), in a greenhouse. The morphometric variables of plant height, diameter at stem height, number of leaves and leaflets, root length and volume, plant dry mass and leaf area were evaluated. The potential evapotranspiration of seedlings (ETc) was obtained using direct weighing, considering as a reference the water replacement of 100% of the mass variation between subsequent days; the cultivation coefficients (kc) were obtained by the ratio between ETc and the reference evapotranspiration (ETo) obtained by the Penman-Montheit FAO-56 method. Biomass and evapotranspiration data were combined to determine water sensitivity. Diurnal fluxes of gas exchange (net photosynthesis rate, transpiration rate, stomatal conductance, internal and atmospheric carbon ratio, water use efficiency and leaf temperature) and Chl a fluorescence (Fv/Fm, ΦPSII, ETR, Fv’/Fm’, NPQ and qL) were evaluated. Water restriction caused reductions of 90.9 and 84.7% in the increase in height and diameter of seedlings subjected to 25% water replacement when compared to seedlings with 100% water replacement, while biomass accumulation reduced by 96.9%. The kc values increased throughout the seedling production cycle, ranging from 0.59 to 2.86. Maximum water sensitivity occurred at 50% water replacement, with Ky = 1.62. Maximum carbon assimilation rates occurred in the morning, ranging from 6.11 to 12.50 µmol m-2 s-1. Ipê-rosa seedlings regulate the physiology of growth, gas exchange, and Chl a fluorescence depending on the amount of water available, and only 25% of the water re-placement in the substrate allows the seedlings to survive.

Harmful algal blooms are a serious problem for humanity, so the development of tools and methods for their timely detection is an urgent task. The use of optical methods often allows scientists to solve problems of remote and automated monitoring. Temperature spectra of fluorescence can be used as distinctive features of algae, and the fluorescence parameters obtained from them can be used for identification. The paper presents a description of the fluorescence parameters of Alexandrium catenella, and shows their stability during cumulative cultivation. In addition, the cytometric properties of the culture and its pigment composition were studied. In addition, the article compares all the studied properties of this microalga with the alga Prorocentrum cordatum. We demonstrated clear differences in their fluorescence parameters, which makes it possible to distinguish these microalgae by optical methods, for example, during their dominant blooms.

Background: Helicobacter pylori (H. pylori) is a prevalent bacterial pathogen implicated in various gastroduodenal diseases, including chronic gastritis, peptic ulcer disease, and gastric cancer. The COVID-19 pandemic has significantly influenced antibiotic prescription practices, potentially exacerbating antibiotic resistance in H. pylori. Objective: This study aims to investigate antibiotic prescription during COVID-19 pandemic in Croatia and its possible impact on H. pylori antibiotic resistance, with a focus on Croatia. Methods: A comprehensive literature search was conducted to identify relevant studies from March 2020 to June 2024. The search strategy included terms related to COVID-19, antibiotic prescription, and Croatia. Studies were selected based on predefined eligibility criteria, focusing on observational research addressing antibiotic use during the pandemic. Results: The COVID-19 pandemic leads to significant fluctuations in community antibiotic consumption within the European Union/European Economic Area. In Croatia, antibiotic use in 2022 surpassed 2019 levels, driven by increased consumption of macrolides and other antibiotics. This surge, influenced by early pandemic treatment claims and penicillin shortages, contributed to rising antibiotic resistance in H. pylori. Resistance rates to clarithromycin and levofloxacin were notably high, driven by mutations in the 23S rRNA, gyrA, and gyrB genes. Discussion: The increased antibiotic use during the COVID-19 pandemic has highly likely complicated H. pylori eradication efforts, highlighting the necessity of judicious antibiotic use and robust antimicrobial stewardship. The pandemic underscored the need for new therapeutic strategies, optimized eradication regimens, and advanced diagnostic methods to manage bacterial infections effectively. Conclusions: The COVID-19 pandemic has significantly impacted antibiotic use and resistance patterns, posing new challenges for H. pylori eradication. Addressing these challenges requires a multifaceted approach, including the development of new drugs and advanced diagnostics, coupled with sustained efforts in antimicrobial stewardship to combat emerging resistance threats.

Candida albicans is present in oral health and has been linked with dental caries. So far, the behavior of the yeast in the presence of Streptococcus sanguinis has not been tested. The aim was to determine the effect of excess sugar as a cariogenic environment on the biological and physico-chemical parameters of single-species (C. albicans) and dual-species biofilms of C. albicans and S. sanguinis. An in vitro caries model was used. The biofilms were exposed to 10% sucrose or 0.9% NaCl for 5 min 3x/day/5 days. Viable cells, hyphal quantification, SEM, biofilm pH, H2O2 quantification and surface microhardness of enamel slabs were determined. Microbial viability and the number of hyphae were higher for sucrose exposure, in the dual-species biofilms induced by excess sugar and coexistence between the two microorganisms. Sucrose-exposed biofilms showed a progressive pH drop over the time course. H2O2 production was about 6 times higher for the dual-species biofilms. Enamel demineralization was higher in the single-species, suggesting an inhibitory effect due to S. sanguinis. Excess sugar increases the pathogenic potential of C. albicans biofilm. In dual-species biofilms, S. sanguinis prevents pronounced drops in pH, decreasing demineralization and stimulates the development of hyphae in the yeast.

BReast CAncer gene 1 (BRCA1) and BReast CAncer gene 2 (BRCA2) encode for tumor suppressor proteins which are critical regulators of the homologous recombination (HR) pathway, the most precise and important DNA damage response mechanism. Dysfunctional HR proteins cannot repair double-stranded DNA breaks in mammalian cells, a situation called HR deficiency. Since their identification, pathogenic variants and other alterations of BRCA1 and BRCA2 genes have been associated with an increased risk of developing mainly breast and ovarian cancer. Interestingly, HR deficiency is also detected in tumors not carrying BRCA1/2 mutations, a condition termed “BRCAness”. One of the main mechanisms causing the BRCAness phenotype is the methylation of the BRCA1/2 promoters and this epigenetic modification is associated with carcinogenesis and poor prognosis mainly among patients with breast and ovarian cancer. BRCA1 promoter methylation has been suggested as an emerging biomarker of great predictive significance, especially concerning Poly (ADP-ribose) Polymerase inhibitors (PARP inhibitor-PARPi) responsiveness, along with or beyond BRCA1/2 mutations. However, as its clinical exploitation is still insufficient, the impact of BRCA1/2 promoter methylation status needs to be further evaluated. The current review aims to gather the latest findings about the mechanisms that underline BRCA1/2 function as well as the molecular characteristics of tumors associated with BRCA1/2 defects, by focusing on DNA methylation. Furthermore, we critically analyze their translational meaning and the validity of BRCA methylation biomarkers in predicting treatment response and we suggest a diagnostic pipeline that could be implemented in liquid biopsy to aid precision pharmacotherapy in BRCA-associated tumors.

Inula viscosa is a widely used plant in traditional Mediterranean and Middle Eastern medicine for various illnesses. I. viscosa has been shown to have anticancerous effects against various cancers, but its effects against lung cancer have been under limited investigation. At the same time, I. viscosa is rich in terpenoids whose anti-lung cancer effects have been poorly investigated. This study aimed to examine the potential anticancerous properties of methanolic and aqueous extracts of stems and leaves of I. viscosa and its terpenoid-rich fraction against human lung cancer A549 cells. Results showed a high radical scavenging capacity and polyphenol and flavonoid contents of I. viscosa methanolic rather than aqueous extracts. In addition, leaves methanolic extracts (IVLM) showed the highest reduction in viability of A549 cells among all the extracts. IVLM also reduced the viability of human ovarian SK-OV-3, breast MCF-7, liver HepG2, and colorectal HCT116 cancer cells. A terpenoid-rich I. viscosa fraction (IVL DCM), prepared by liquid-liquid separation of IVLM in dichloromethane (DCM), displayed a substantial reduction in the viability of A549 cells (IC₅₀= 27.8 ± 1.5 µg/mL at 48 h) and the panel of tested cancerous cell lines but was not cytotoxic to normal human embryonic fibroblasts (HDFn). The assessment of IVL DCM phytochemical constituents using GC-MS analysis revealed 21 metabolites, highlighting an enrichment in terpenoids, such as lupeol and its derivatives, caryophyllene oxide, betulin, and isopulegol, known to exhibit proapoptotic, and antimetastatic functions. IVL DCM also showed robust antioxidant capacity and decent polyphenol and flavonoid contents. Furthermore, Western blotting analysis indicated that IVL DCM reduced proliferation (reduction of proliferation marker Ki67 and induction of proliferation inhibitor proteins P21 and P27), contaminant with P38 MAP kinase activation, and induced the intrinsic apoptotic pathway (P53/BCL2/BAX/ Caspase3/PARP) in A549 cells. IVL DCM also reduced the migration of A549 cells, potentially by reducing FAK activation. Future identification of anticancer metabolites of IVL DCM, especially terpenoids, is recommended. These data place I. viscosa as a new resource of herbal anticancer agents.