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19 pages, 3747 KiB  
Article
Furostanol Saponins and Ecdysteroids from Plants of the Genus Helleborus as Phagostimulants and Predator Deterrents for Larvae of Two Monophadnus Sawfly Species
by Jose M. Prieto, Alison M. Barker, Urs Schaffner, Joëlle Quetin-Leclercq, Alessandra Braca and Jean-Luc Boevé
Plants 2024, 13(16), 2230; https://doi.org/10.3390/plants13162230 (registering DOI) - 12 Aug 2024
Abstract
Sawfly species of the genus Monophadnus are specialised on Ranunculaceae plants from which the larvae can sequester furostanol saponins into the haemolymph, mainly (25R)-26-[(α-L-rhamnopyranosyl)oxy]-22α-methoxyfurost-5-en-3β-yl-O-β-D-glucopyranosyl-(1→3)-O-[6-acetyl-β-D-glucopyranosyl-(1→3)]-O-β-D-glucopyranoside (compound 1). In this work, TLC, GC-MS, and HPLC-DAD-ESI/MS analyses [...] Read more.
Sawfly species of the genus Monophadnus are specialised on Ranunculaceae plants from which the larvae can sequester furostanol saponins into the haemolymph, mainly (25R)-26-[(α-L-rhamnopyranosyl)oxy]-22α-methoxyfurost-5-en-3β-yl-O-β-D-glucopyranosyl-(1→3)-O-[6-acetyl-β-D-glucopyranosyl-(1→3)]-O-β-D-glucopyranoside (compound 1). In this work, TLC, GC-MS, and HPLC-DAD-ESI/MS analyses together with feeding, repeated simulated attacks, and ant deterrence bioassays were conducted to extend the chemoecological knowledge about two sawfly species specialised on H. foetidus L. (Monophadnus species A) and H. viridis L. (Monophadnus species B). Larvae of Monophadnus species B were mostly feeding on the squares treated with the n-butanol fraction from H. foetidus, compound 1 being its primary non-nutritional stimulant. In contrast, all H. viridis fractions stimulated feeding, with n-hexane marginally more active. β-sitosterol within n-hexane was determined as the nutritional stimulant. Quantitative analyses demonstrated that leaves of H. viridis but not H. foetidus contain the ecdysteroids 20-hydroxyecdysone and polypodine B. Moreover, the haemolymph of Monophadnus species B larvae reared on H. viridis contained the glycosides of polypodine B and 20-hydroxyecdysone at a concentration of 2.5 to 6.8 µmol/g fresh weight of haemolymph. This concentration is several thousand times higher than the concentration range of the aglycones in their host plant (3.63 × 10−4 to 2.23 × 10−4 µmol total ecdysteroids/g fresh weight of leaves), suggesting bioaccumulation. The larvae of both species fed on H. foetidus do not show any traces of ecdysteroids in their haemolymph, indicating a facultative role of these compounds in their defence as well as their inability to endogenously synthesise these compounds. The haemolymph containing ecdysteroids was a significant feeding deterrent against Myrmica rubra L. ant workers (one of their natural predators) at 0.8 mg/mL. The larvae kept effective deterrent levels of glycosylated ecdysteroids (≅175 mM) between simulated attacks on days 1 and 2, but the levels clearly decreased on day 3 (≅75 mM). Most larvae (89%) survived a first attack but only 23% a consecutive second one. As a conclusion, we report for the first time that two Monophadnus species feeding on H. viridis sequester phytoecdysteroids into the larval haemolymph in the form of glycosides. In addition, compound 1 possesses defensive and phagostimulant activities, and we present evidence for a combined effect of furostanol saponins and ecdysteroids as repellents against ants. Full article
(This article belongs to the Special Issue Medicinal Plants: Advances in Phytochemistry and Ethnobotany II)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Larvae of <span class="html-italic">Monophadnus</span> species A (<b>left</b>, picture by Alison M. Barker) and <span class="html-italic">Monophadnus</span> species B (<b>right</b>, picture by Jean-Luc Boevé) attached to the underside of a leaf of <span class="html-italic">Helleborus foetidus</span> L. and <span class="html-italic">Helleborus viridis</span> L., respectively.</p>
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<p>Steroidal furostanol saponin (25<span class="html-italic">R</span>)-26-[(α-L-rhamnopyranosyl)oxy]-22α-methoxyfurost-5-en-3β-yl-<span class="html-italic">O</span>-β-D-glucopyranosyl-(1→3)-<span class="html-italic">O</span>-[6-acetyl-β-D-glucopyranosyl-(1→3)]-<span class="html-italic">O</span>-β-D-glucopyranoside (compound <b>1</b>) found in <span class="html-italic">H. viridis</span>, <span class="html-italic">H. foetidus</span>, and the haemolymph of the two <span class="html-italic">Monophadnus</span> spp. that feed on them.</p>
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<p>Ranunculin (compound <b>2</b>) and its derivatives protoanemonin (compound <b>3</b>) and anemonin (compound <b>4</b>) are characteristic g-lactones found in <span class="html-italic">R</span>-type Ranunculaceae.</p>
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<p>The ecdysteroids 20-hydroxyecdysone (R = H) (compound <b>5</b>) and polypodine B (R = OH) (compound <b>6</b>) are naturally occurring in <span class="html-italic">Helleborus viridis</span> but not in <span class="html-italic">Helleborus foetidus</span>.</p>
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<p>Feeding bioassays of <span class="html-italic">Monophadnus</span> species B on paper squares treated with the initial crude methanol extract and its increasing polarity solvent fractions (<b>a</b>,<b>b</b>) and selected subfractions of the active <span class="html-italic">n</span>-butanol fraction of <span class="html-italic">Helleborus foetidus</span> (<b>c</b>) and the <span class="html-italic">n</span>-hexane fraction of <span class="html-italic">Helleborus viridis</span> (<b>d</b>) leaves. Orange and yellow colours indicate larvae feeding on fraction-treated and solvent control squares, respectively, at 6 h. (Pig) Pigments. (*) <span class="html-italic">p</span> &lt; 0.05; (**) <span class="html-italic">p</span> &lt; 0.01, Wilcoxon signed-rank tests.</p>
Full article ">Figure 6
<p>Feeding bioassay of <span class="html-italic">Monophadnus</span> species A on paper squares treated with 0.5 mg/mL of compound <b>1</b>. (**) <span class="html-italic">p</span> &lt; 0.01 and F-distribution were calculated according Hotelling’s T<sup>2</sup> test. Total weight of paper targets available was on average 80 mg per replicate (40 mg controls, 40 mg treatments), so larvae still had both choices available at the end of the experiment.</p>
Full article ">Figure 7
<p>HPLC/MS chromatograms of <span class="html-italic">Helleborus</span> spp. <span class="html-italic">n</span>-butanol extracts, haemolymph of <span class="html-italic">Monophadnus</span> species B larvae feeding on <span class="html-italic">Helleborus viridis</span>, and haemolymph of <span class="html-italic">Monophadnus</span> species A larvae feeding on <span class="html-italic">Helleborus foetidus</span>.</p>
Full article ">Figure 8
<p>Annotated MS spectra of the ecdysteroids detected in the haemolymph of <span class="html-italic">Monophadnus</span> species B feeding on <span class="html-italic">Helleborus viridis</span>.</p>
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<p>TLC analysis (mobile phase: BAW; reagent: cerium sulphate; picture taken under 254 nm UV light in inversed B/W for visual enhancement) of the haemolymph of <span class="html-italic">Monophadnus</span> species A (lanes denoted after sample codes 7 and 8; see <a href="#sec4dot1-plants-13-02230" class="html-sec">Section 4.1</a>) and species B (lanes denoted after sample codes 3, 4, 11, and 12; see <a href="#sec4dot1-plants-13-02230" class="html-sec">Section 4.1</a>) reared either in <span class="html-italic">Helleborus viridis</span> or <span class="html-italic">Helleborus foetidus</span>. The yellow circles highlight areas where ecdysteroid metabolites can be detected. The small numbers between dashed lines correspond to different metabolites found in that zone.</p>
Full article ">
14 pages, 2825 KiB  
Article
Method of Planning and Scheduling the Production Process of Yellow Mealworm Larvae for a Small Enterprise
by Arkadiusz Kowalski
Appl. Sci. 2024, 14(16), 7051; https://doi.org/10.3390/app14167051 (registering DOI) - 12 Aug 2024
Abstract
In the context of the growing demand for alternative protein sources with the growth of the human population and increasing ecological awareness, the rearing of yellow mealworm larvae (Tenebrio molitor) is a promising option for the production of sustainable protein. The [...] Read more.
In the context of the growing demand for alternative protein sources with the growth of the human population and increasing ecological awareness, the rearing of yellow mealworm larvae (Tenebrio molitor) is a promising option for the production of sustainable protein. The article presents a comprehensive approach to planning and scheduling the production of yellow mealworm larvae in a small enterprise, focusing on the organizational, technical, and economic aspects of the production process. The production installation, the method of rearing using an automated feeding system, and the monitoring of larvae development were described and an attempt was made to identify the key parameters of the process that affect its efficiency. Particular attention was paid to the calculation algorithm implemented in the spreadsheet, which allows the selection of the production batch size and the frequency of their launch, so as to maximize the available capacity of storage racks for cuvettes. In addition, the article analyses logistical challenges related to the production of larvae, including transport activities in order to meet, among others, the demand for feed. Finally, the estimation of revenues and economic indicators, such as profitability and return on investment, is presented, pointing to the need for further improvements in the production process and cost optimization to achieve favorable financial results. The results of the research emphasize the potential of rearing yellow mealworm larvae as a sustainable source of protein while simultaneously pointing to key areas that require further research and development. Full article
(This article belongs to the Special Issue Sustainable and Smart Agriculture)
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Figure 1

Figure 1
<p>The proposed shape of the production process of yellow mealworm larvae is divided into “larger” and “smaller” larvae fractions and “extra fattening”.</p>
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<p>The proposed three-level method of yellow mealworm larvae feeding.</p>
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<p>The growth rate of yellow mealworm larvae of various size fractions for the adopted feeding method.</p>
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<p>The idea of building a production plan considering the timeline.</p>
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<p>Schematic diagram of calculations for the production schedule and plan.</p>
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<p>Determination of the maximum number of occupied cuvettes when starting full-scale production.</p>
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<p>Weight of harvesting larvae when starting subsequent batches of 220 cuvettes every 7 days.</p>
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12 pages, 1225 KiB  
Article
Macroinvertebrates Associated with Macroalgae within Integrated Multi-Trophic Aquaculture (IMTA) in Earthen Ponds: Potential for Accessory Production
by Rafael Vieira, Miguel Ângelo Mateus, Carlos Manuel Lourenço Afonso, Florbela Soares, Pedro Pousão-Ferreira and Sofia Gamito
J. Mar. Sci. Eng. 2024, 12(8), 1369; https://doi.org/10.3390/jmse12081369 (registering DOI) - 11 Aug 2024
Abstract
The present work aims to evaluate the macroinvertebrate community associated with macroalgae in earthen pond systems to better understand their potential in detritus recycling and as an accessory production. Sampling took place on the settling pond of an aquaculture research station, where macroalgae [...] Read more.
The present work aims to evaluate the macroinvertebrate community associated with macroalgae in earthen pond systems to better understand their potential in detritus recycling and as an accessory production. Sampling took place on the settling pond of an aquaculture research station, where macroalgae permanently occurred at high densities. The results suggest differentiation between seasons but not between sites within the settling pond. Seasonal variation was observable in terms of macroinvertebrate density, biomass, and diversity. Two non-indigenous species of invertebrates were found, the crustaceans Grandidierella japonica and Paracerceis sculpta Amphipods were the most abundant group, and their high nutritional value can be exploited. Detritus and the epiphyte layer are the main food items for the invertebrates, reinforcing the advantages of these organisms being present to enhance the recycling of excess detritus and to transfer organic matter to upper trophic levels. These species, naturally present in aquaculture facilities, can improve the water quality and increase the variability of food nutrients for reared species. Full article
(This article belongs to the Topic Aquatic Environment Research for Sustainable Development)
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Figure 1

Figure 1
<p>Seasonal variation plots with standard error for (<b>a</b>) dried algae biomass by cubic metre of water; (<b>b</b>) invertebrates’ biomass by cubic metre of water; (<b>c</b>) invertebrates’ density by gram of algae dry weight; (<b>d</b>) invertebrates’ density by cubic metre of water.</p>
Full article ">Figure 2
<p>Scatter plots of taxa richness (<b>A</b>) and number of individuals (<b>B</b>) per algae dry weight (DW) based on season. A: Autumn (Y = 0.1608x + 5.030); Winter (Y = 0.3252x + 4.921); Spring (Y = 0.2446x + 12.96). B: Autumn (Y = 6.681x + 196.0); Winter (Y = 0.7874x + 582.9); Spring (Y = 166.2x − 565.5).</p>
Full article ">Figure 3
<p>Variation, per season, in the six most abundant species (<span class="html-italic">Monocorophium insidiosum</span>, <span class="html-italic">Hydrobia glyca</span>, <span class="html-italic">Paracerceis sculpta</span>, <span class="html-italic">Grandidierella japonica</span>, <span class="html-italic">Cymadusa filose</span> and <span class="html-italic">Microdeutopus gryllotalpa</span>). (<b>a</b>) Density per gram of dry algae and (<b>b</b>) density per cubic metre.</p>
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<p>Multidimensional scaling (MDS) carried out with macroinvertebrates associated with a macroalgae abundance dataset using Bray–Curtis similarity index and root transformation. Four samples were collected for each season: Autumn (Aut1–4), Winter (Win1–4), and Spring (Spr1–4).</p>
Full article ">Figure 5
<p>(<b>a</b>)Variation per season in the main taxonomic groups; (<b>b</b>) density of the most abundant benthic macroinvertebrates in Autumn sampling (<span class="html-italic">Capitella</span> sp., <span class="html-italic">Neanthes acuminata</span>, <span class="html-italic">Monocorophium insidiosum</span>, <span class="html-italic">Gammarella fucicola</span>, <span class="html-italic">Grandidierella japonica</span>, <span class="html-italic">Hydrobia glyca,</span> and <span class="html-italic">Peringia ulvae</span>).</p>
Full article ">
12 pages, 468 KiB  
Article
Effect of Adding the Antimicrobial L-Carnitine to Growing Rabbits’ Drinking Water on Growth Efficiency, Hematological, Biochemical, and Carcass Aspects
by Mohamed I. Hassan, Naela Abdel-Monem, Ayman Moawed Khalifah, Saber S. Hassan, Hossam Shahba, Ahmad R. Alhimaidi, In Ho Kim and Hossam M. El-Tahan
Antibiotics 2024, 13(8), 757; https://doi.org/10.3390/antibiotics13080757 (registering DOI) - 11 Aug 2024
Abstract
The current study was designed to assess the impact of L-carnitine (LC) supplementation in the drinking water of growing Alexandria-line rabbits on performance and physiological parameters. Two hundred eighty-eight 35-day-old rabbits were divided into four groups of twenty-four replicates each (seventy-two rabbits/treatment). The [...] Read more.
The current study was designed to assess the impact of L-carnitine (LC) supplementation in the drinking water of growing Alexandria-line rabbits on performance and physiological parameters. Two hundred eighty-eight 35-day-old rabbits were divided into four groups of twenty-four replicates each (seventy-two rabbits/treatment). The treatment groups were a control group without LC and three groups receiving 0.5, 1, and 1.5 g/L LC in the drinking water intermittently. The results showed that the group receiving 0.5 g LC/L exhibited significant improvements in final body weight, body weight gain, feed conversion ratio, and performance index compared to the other groups. The feed intake remained unaffected except for the 1.5 g LC/L group, which had significantly decreased intake. Hematological parameters improved in all supplemented groups. Compared with those in the control group, the 0.5 g LC/L group showed significant increases in serum total protein and high-density lipoprotein, along with decreased cholesterol and low-density lipoprotein. Compared to other supplemented groups, this group also demonstrated superior carcass traits (carcass, dressing, giblets, and percentage of nonedible parts). In conclusion, intermittent supplementation of LC in the drinking water, particularly at 0.5 g/L twice a week, positively influenced the productivity, hematology, serum lipid profile, and carcass traits of Alexandria-line growing rabbits at 84 days of age. Full article
(This article belongs to the Special Issue Natural Compounds as Antimicrobial Agents, 2nd Edition)
24 pages, 9689 KiB  
Article
Genome-Wide Identification, Evolution, and Expression Analysis of the Dirigent Gene Family in Cassava (Manihot esculenta Crantz)
by Mingchao Li, Kai Luo, Wenke Zhang, Man Liu, Yunfei Zhang, Huling Huang, Yinhua Chen, Shugao Fan and Rui Zhang
Agronomy 2024, 14(8), 1758; https://doi.org/10.3390/agronomy14081758 (registering DOI) - 11 Aug 2024
Abstract
Dirigent (DIR) genes play a pivotal role in plant development and stress adaptation. Manihot esculenta Crantz, commonly known as cassava, is a drought-resistant plant thriving in tropical and subtropical areas. It is extensively utilized for starch production, bioethanol, and animal feed. [...] Read more.
Dirigent (DIR) genes play a pivotal role in plant development and stress adaptation. Manihot esculenta Crantz, commonly known as cassava, is a drought-resistant plant thriving in tropical and subtropical areas. It is extensively utilized for starch production, bioethanol, and animal feed. However, a comprehensive analysis of the DIR family genes remains unexplored in cassava, a crucial cash and forage crop in tropical and subtropical regions. In this study, we characterize a total of 26 cassava DIRs (MeDIRs) within the cassava genome, revealing their uneven distribution across 13 of the 18 chromosomes. Phylogenetic analysis classified these genes into four subfamilies: DIR-a, DIR-b/d, DIR-c, and DIR-e. Comparative synteny analysis with cassava and seven other plant species (Arabidopsis (Arabidopsis thaliana), poplar (Populus trichocarpa), soybean (Glycine max), tomato (Solanum lycopersicum), rice (Oryza sativa), maize (Zea mays), and wheat (Triticum aestivum)) provided insights into their likely evolution. We also predict protein interaction networks and identify cis-acting elements, elucidating the functional differences in MeDIR genes. Notably, MeDIR genes exhibited specific expression patterns across different tissues and in response to various abiotic and biotic stressors, such as pathogenic bacteria, cadmium chloride (CdCl2), and atrazine. Further validation through quantitative real-time PCR (qRT-PCR) confirmed the response of DIR genes to osmotic and salt stress. These findings offer a comprehensive resource for understanding the characteristics and biological functions of MeDIR genes in cassava, enhancing our knowledge of plant stress adaptation mechanisms. Full article
22 pages, 2478 KiB  
Article
StreetLines: A Smart and Scalable Tourism Platform Based on Efficient Knowledge-Mining
by Georgios Alexandridis, Georgios Siolas, Tasos Papagiannis, George Ioannou, Konstantinos Michalakis, George Caridakis, Vasileios Karyotis and Symeon Papavassiliou
Digital 2024, 4(3), 676-697; https://doi.org/10.3390/digital4030034 (registering DOI) - 11 Aug 2024
Abstract
Identifying and understanding visitor needs and expectations is of the utmost importance for a number of stakeholders and policymakers involved in the touristic domain. Apart from traditional forms of feedback, an abundance of related information exists online, scattered across various data sources like [...] Read more.
Identifying and understanding visitor needs and expectations is of the utmost importance for a number of stakeholders and policymakers involved in the touristic domain. Apart from traditional forms of feedback, an abundance of related information exists online, scattered across various data sources like online social media, tourism-related platforms, traveling blogs, forums, etc. Retrieving and analyzing the aforementioned content is not a straightforward task and in order to address this challenge, we have developed the StreetLines platform, a novel information system that is able to collect, analyze and produce insights from the available tourism-related data. Its highly modular architecture allows for the continuous monitoring of varying pools of heterogeneous data sources whose contents are subsequently stored, after preprocessing, in a data repository. Following that, the aforementioned data feed a number of independent and parallel processing modules that extract useful information for all individuals involved in the tourism domain, like place recommendation for visitors and sentiment analysis and keyword extraction reports for professionals in the tourism industry. The presented platform is an outcome of the StreetLines project and apart from the contributions of its individual components, its novelty lies in the holistic approach to knowledge extraction and tourism data mining. Full article
(This article belongs to the Collection Digital Systems for Tourism)
17 pages, 3553 KiB  
Article
Comparative Transcriptome Analysis Unveils Regulatory Factors Influencing Fatty Liver Development in Lion-Head Geese under High-Intake Feeding Compared to Normal Feeding
by Jie Kong, Ziqi Yao, Junpeng Chen, Qiqi Zhao, Tong Li, Mengyue Dong, Yuhang Bai, Yuanjia Liu, Zhenping Lin, Qingmei Xie and Xinheng Zhang
Vet. Sci. 2024, 11(8), 366; https://doi.org/10.3390/vetsci11080366 (registering DOI) - 11 Aug 2024
Viewed by 80
Abstract
The lion-head goose is the only large goose species in China, and it is one of the largest goose species in the world. Lion-head geese have a strong tolerance for massive energy intake and show a priority of fat accumulation in liver tissue [...] Read more.
The lion-head goose is the only large goose species in China, and it is one of the largest goose species in the world. Lion-head geese have a strong tolerance for massive energy intake and show a priority of fat accumulation in liver tissue through special feeding. Therefore, the aim of this study was to investigate the impact of high feed intake compared to normal feeding conditions on the transcriptome changes associated with fatty liver development in lion-head geese. In this study, 20 healthy adult lion-head geese were randomly assigned to a control group (CONTROL, n = 10) and high-intake-fed group (CASE, n = 10). After 38 d of treatment, all geese were sacrificed, and liver samples were collected. Three geese were randomly selected from the CONTROL and CASE groups, respectively, to perform whole-transcriptome analysis to analyze the key regulatory genes. We identified 716 differentially expressed mRNAs, 145 differentially expressed circRNAs, and 39 differentially expressed lncRNAs, including upregulated and downregulated genes. GO enrichment analysis showed that these genes were significantly enriched in molecular function. The node degree analysis and centrality metrics of the mRNA–lncRNA–circRNA triple regulatory network indicate the presence of crucial functional nodes in the network. We identified differentially expressed genes, including HSPB9, Pgk1, Hsp70, ME2, malic enzyme, HSP90, FADS1, transferrin, FABP, PKM2, Serpin2, and PKS, and we additionally confirmed the accuracy of sequencing at the RNA level. In this study, we studied for the first time the important differential genes that regulate fatty liver in high-intake feeding of the lion-head goose. In summary, these differentially expressed genes may play important roles in fatty liver development in the lion-head goose, and the functions and mechanisms should be investigated in future studies. Full article
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Figure 1

Figure 1
<p>Phenotypic changes in liver and body weight after high-intake feeding in lion-head geese. (<b>A</b>) Goose liver in the groups of high-intake feeding and normal feeding. (<b>B</b>) The weight of liver. (<b>C</b>) The size of the goose liver. (<b>D</b>) The weight of goose (** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Overview of whole-transcriptome sequencing. (<b>A</b>) The density distribution of transcriptome was according to log10 (FPKM), original gene read counts were normalized using the FPKM method. (<b>B</b>) The 6−sample expression violin plot. (<b>C</b>) The principal component analysis (PCA) is useful for exploring the distance relationship between the 6 samples. (<b>D</b>) Pearson’s correlation matrix for mRNA, lncRNA, and circRNA profiles.</p>
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<p>Identification and analysis of mRNA, lncRNA, and circRNA associated with fatty liver. (<b>A</b>) Volcano plots of DEGs in mRNA. (<b>B</b>) Volcano plots of DEGs in lncRNA. (<b>C</b>) Volcano plots of DEGs in circRNA. Each dot in the plot represents a gene with its corresponding log2 fold change (FC) on the x-axis and <span class="html-italic">p</span>-value (log10) on the y-axis. The horizontal line indicates the significance threshold (false discovery rate &lt;5%), whereas the vertical line segregates genes with logFC &gt;1.5. Gray represents no significant difference between the two groups, red indicates the upregulated genes, and blue indicates the downregulated genes.</p>
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<p>Functional analyses of significantly enriched trends. The top 8 significant terms at the mRNA level. The size of dots shows the number of DEGs clustered in the same terms. The color of dots indicates the <span class="html-italic">p</span>-value.</p>
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<p>Construction and visualization of lncRNA–mRNA–circRNA network. (<b>A</b>) lncRNA–mRNA–circRNA network structure. (<b>B</b>) The betweenness centrality difference among lncRNAs, mRNAs, and circRNAs. (<b>C</b>) The closeness centrality difference among lncRNAs, mRNAs, and circRNAs. (<b>D</b>) The degree centrality difference among lncRNAs, mRNAs, and circRNAs. (<b>E</b>) Node degree of the lncRNAs, mRNAs, and circRNAs.</p>
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<p>Comparison and identification of differentially expressed RNAs in high-intake feeding in lion-head geese liver tissue by qRT-PCR. (<b>A</b>) The RNA-seq results revealed the upregulated genes by FPKM. (<b>B</b>) Verification of the upregulated genes by RT-qPCR. (<b>C</b>) The RNA-seq results revealed the downregulated genes by FPKM. (<b>D</b>) Verification of the downregulated genes by RT-qPCR. Log2 FC is expressed as mean ± SD. n = 3. The statistical significance of all genes reached <span class="html-italic">p</span> &lt; 0.05 (*** <span class="html-italic">p</span> &lt; 0.001; ** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05).</p>
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18 pages, 2668 KiB  
Article
Innovative Foraging Behavior of Urban Birds: Use of Insect Food Provided by Cars
by Jukka Jokimäki and Anna Ramos-Chernenko
Birds 2024, 5(3), 469-486; https://doi.org/10.3390/birds5030032 (registering DOI) - 10 Aug 2024
Viewed by 314
Abstract
Despite high-quality insect food being often restricted in cities, insects are important for the development of birds. Nonetheless, plenty of insects are smashed on cars, and they are available for those species that are able to use them. We used both our own [...] Read more.
Despite high-quality insect food being often restricted in cities, insects are important for the development of birds. Nonetheless, plenty of insects are smashed on cars, and they are available for those species that are able to use them. We used both our own data and community science and Internet sources for surveying global, national, and local data about birds using insects on cars. Our results contained a total of 309 observations of birds collecting insects on car panels, which indicated that 39 species used this food resource since 1928 in 31 countries. Most observations considered the House Sparrow, followed by the White Wagtail and several species of corvids. European urban bird species observed to use insects on cars had a larger residual brain size. There was also some indication that bird species using insects on cars had a larger number of innovations (i.e., production of novel behaviors), greater diet generalism, and longer times living in urbanized areas than birds not observed using insects on cars. Often these species are also resident and able to use food offered in feeding sites. We assume that more bird species will use insects on cars in the future, as urban insect populations continue to decline, and thereby insects on cars will increasingly become more important sources of food for urban birds. Full article
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<p>Country-based distribution and numbers of the observations of (<b>A</b>) House Sparrows (<span class="html-italic">Passer domesticus</span>), (<b>B</b>) White Wagtails (<span class="html-italic">Motacilla alba</span>), (<b>C</b>) Eurasian Jackdaws (<span class="html-italic">Corvus monedula</span>), (<b>D</b>) Carrion/Hooded Crows (<span class="html-italic">Corvus corone</span> and <span class="html-italic">Corvus cornix</span>, respectively) picking insects from cars, and (<b>E</b>) countries (marked in green with a green dot) were the responders reported that they have not observed such behavior.</p>
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<p>Proportion (%) of species of the total number of foraging observations (<span class="html-italic">n</span> = 61; blue bars) and of the total numbers of birds observed (<span class="html-italic">n</span> = 477; green bars) in the urban parking areas in Rovaniemi, Finland. Cmon = <span class="html-italic">Corvus monedula</span>, Cnix = <span class="html-italic">Corvus cornix</span>, Lrid = <span class="html-italic">Larus ridibundus</span>, Malb = <span class="html-italic">Motacilla alba</span>, Pdom = <span class="html-italic">Passer domesticus</span>, Pmaj = <span class="html-italic">Parus major</span>, Ppic = <span class="html-italic">Pica pica</span>, and other = sum of the following other species (<span class="html-italic">Apus apus</span>, <span class="html-italic">Carduelis chloris</span>, <span class="html-italic">Cyanistes caeruleus</span>, <span class="html-italic">Columba livia</span>, <span class="html-italic">Corvus corax</span>, <span class="html-italic">Delichon urbica</span>, <span class="html-italic">Fringilla coelebs</span>, <span class="html-italic">Hirundo rustica</span>, <span class="html-italic">Larus canus</span>, <span class="html-italic">Motacilla flava</span>, <span class="html-italic">Passer montanus</span>, <span class="html-italic">Turdus iliacus</span>, and <span class="html-italic">Turdus pilaris</span>).</p>
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17 pages, 1063 KiB  
Article
Intestinal Microbiome Profiles in Broiler Chickens Raised with Different Probiotic Strains
by Julia Marixara Sousa da Silva, Ana Maria De Souza Almeida, Ana Carolina Borsanelli, Flávia Regina Florencio de Athayde, Eduardo de Paula Nascente, João Marcos Monteiro Batista, Alison Batista Vieira Silva Gouveia, José Henrique Stringhini, Nadja Susana Mogyca Leandro and Marcos Barcellos Café
Microorganisms 2024, 12(8), 1639; https://doi.org/10.3390/microorganisms12081639 (registering DOI) - 10 Aug 2024
Viewed by 258
Abstract
The composition of the intestinal microbiota can influence the metabolism and overall functioning of avian organisms. Therefore, the objective of this study was to evaluate the effect of three different probiotics and an antibiotic on the microbiomes of 1.400 male Cobb® broiler [...] Read more.
The composition of the intestinal microbiota can influence the metabolism and overall functioning of avian organisms. Therefore, the objective of this study was to evaluate the effect of three different probiotics and an antibiotic on the microbiomes of 1.400 male Cobb® broiler raised for 42 days. The experiment was conducted with the following treatments: positive control diet (basal diet + antibiotic); negative control diet (basal diet without antibiotic and without probiotic); basal diet + Normal Avian Gut Flora (NAGF); basal diet + multiple colonizing strain probiotics (MCSPs); and basal diet + non-colonizing single strain probiotics (NCSSPs). The antibiotic (enramycin—antibiotic growth promoter) and probiotics were administered orally during all experiment (1 to 42 days), mixed with broiler feed. To determine the composition of the microbiota, five samples of ileal digesta were collected from 42-day-old chickens of each experimental group. The alpha and beta diversity of the ileal microbiota showed differences between the groups. MCSPs presented greater richness and uniformity compared to the positive control, negative control, and NCSSPs treatments, while the negative control exhibited greater homogeneity among samples than NCSSPs. MCSPs also showed a higher abundance of the genus Enterococcus. There were differences between the groups for low-abundance taxa (<0.5%), with NAGF showing higher levels of Delftia, Brevibacterium, and Bulleidia. In contrast, NCSSPs had a higher abundance of Ochrobactrum, Rhodoplanes, and Nitrospira. It was concluded that the treatments analyzed in this study induced modulations in the ileal microbiota of the chickens examined. Full article
(This article belongs to the Section Veterinary Microbiology)
13 pages, 1470 KiB  
Article
Reproductive Success of Tree Swallows at Abandoned Mine Drainage Treatment Ponds
by James S. Kellam, Julianna E. Lott, Anna R. Doelling and Isabella Ladisic
Birds 2024, 5(3), 440-452; https://doi.org/10.3390/birds5030030 (registering DOI) - 10 Aug 2024
Viewed by 229
Abstract
Abandoned mine drainage treatment ponds could have contrasting effects on the reproductive success of birds living in the vicinity. The ponds and associated vegetation may, like any other body of freshwater, provide beneficial habitats for the insects that the birds use to feed [...] Read more.
Abandoned mine drainage treatment ponds could have contrasting effects on the reproductive success of birds living in the vicinity. The ponds and associated vegetation may, like any other body of freshwater, provide beneficial habitats for the insects that the birds use to feed their young; or instead, the ponds may act as an ecological trap, attracting the birds to a habitat that is poor in quality and negatively impacting their productivity. We monitored nests of an aerial insectivore, the Tree Swallow (Tachycineta bicolor), to determine whether the distance between the ponds and the nests affected various reproductive parameters including clutch size, hatch rate, number of nestlings, nestling size and mass, number of fledglings, fledging rate, and fledge date. Data were collected over two breeding seasons (2022 and 2023) from a swallow population in southwestern Pennsylvania, USA. We found that the nests closest to the treatment ponds had significantly more nestlings and fledglings, earlier fledge dates, and a better fledging rate when compared to nests that were more distant from the ponds. However, all these parameters were well below previously published values, which suggests that the mine drainage ponds provide good nesting habitats relative to what is available in the region but that they do not represent high-quality habitats for this species overall. Full article
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<p>(<b>a</b>) Location of the study site within the United States. (<b>b</b>) Aerial view of the study site. Tree Swallow nest boxes were placed near abandoned mine drainage (AMD) treatment ponds and at locations that were more distant from the ponds. Nest box locations are shown on the map as red squares (NEAR locations), yellow squares (MID locations), and purple squares (FAR locations). Box locations are approximated.</p>
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<p>Average numbers ± SE of Tree Swallow eggs, hatched young, and fledglings at nests NEAR to (red points), at MID distance from (yellow points), and FAR from (purple points) abandoned mine drainage ponds. Overall means of three distance groups were significantly different.</p>
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<p>Wing lengths of nestling Tree Swallows and age at time of measurement. Only one nestling was measured per nest. The positive relationship between the two variables was significant (<span class="html-italic">p</span> &lt; 0.001), with no difference between slopes of nests in the NEAR (red points), MID (yellow points), and FAR (blue points) groups (<span class="html-italic">p</span> = 0.436). Regression lines and 95% confidence intervals are shown for each group. The R<sup>2</sup> values for the regression lines are as follows: NEAR = 0.45, MID = 0.55, FAR = 0.73).</p>
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<p>The mass of nestling Tree Swallows and age at time of measurement. Only one nestling was measured per nest. The positive relationship between the two variables was significant (<span class="html-italic">p</span> = 0.002), with no difference found between nests in the NEAR (red points), MID (yellow points), and FAR (blue points) groups (<span class="html-italic">p</span> = 0.451). Regression lines and 95% confidence intervals are shown for each group. The R<sup>2</sup> values for the regression lines are as follows: NEAR = 0.02, MID = 0.59, FAR = 0.59).</p>
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15 pages, 1248 KiB  
Article
Impact of Incorporating Dried Chaga Mushroom (Inonotus obliquus) into Gluten-Free Bread on Its Antioxidant and Sensory Characteristics
by Zbigniew Kobus, Monika Krzywicka, Agata Blicharz-Kania, Alicja Bosacka, Anna Pecyna, Eva Ivanišová, Katarzyna Kozłowicz and Eva Kovačiková
Molecules 2024, 29(16), 3801; https://doi.org/10.3390/molecules29163801 (registering DOI) - 10 Aug 2024
Viewed by 208
Abstract
Gluten-free bread is increasingly popular among individuals with celiac disease, and The incorporation of mushroom flour offers a novel method to enhance its nutritional profile, antioxidant content, and sensory properties. This study aimed to evaluate the antioxidant and sensory characteristics of gluten-free bread [...] Read more.
Gluten-free bread is increasingly popular among individuals with celiac disease, and The incorporation of mushroom flour offers a novel method to enhance its nutritional profile, antioxidant content, and sensory properties. This study aimed to evaluate the antioxidant and sensory characteristics of gluten-free bread with varying amounts of chaga mushroom flour (5%, 10%, 15%, 20%). The total contents of polyphenols and flavonoids were measured using a spectrophotometric method. Antioxidant activity was assessed through DPPH and FRAP methods, while textural properties were evaluated using the TPA test. Bread colour was analysed using the CIELab system, and sensory evaluation was performed by a panel of trained consumers. The results showed that gluten-free bread enriched with chaga flour had increased polyphenol and flavonoid content and enhanced antioxidant activity. The highest levels of polyphenols, flavonoids, DPPH, and FRAP activity were found in bread with 20% chaga. The addition of chaga mushroom significantly affected the bread’s hardness, cohesiveness, and chewiness. Specifically, 20% chaga flour had the most pronounced effect on hardness and elasticity, while 15% chaga flour had the greatest impact on chewiness and cohesiveness. The bread’s colour darkened with higher chaga concentrations. The results of sensory evaluation showed a negative correlation between consumer preferences and bread fortified with chaga mushroom flour. The overall consumer acceptability score indicates that only a small addition of mushroom flour (up to 10%) can be used to bake gluten-free bread. Full article
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<p>Gluten-free bread with chaga: 0, 5, 10, 15, 20% respectively (from left).</p>
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<p>Sensory evaluation of gluten-free bread with the addition of chaga mushroom.</p>
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10 pages, 776 KiB  
Article
Direct Observation of Feeding Behavior of Adult Tabanidae (Diptera) on Beef Cattle from Khon Kaen Province in Thailand
by Kanchana Thinnabut, Wanchai Maleewong and Ubon Tangkawanit
Insects 2024, 15(8), 602; https://doi.org/10.3390/insects15080602 (registering DOI) - 10 Aug 2024
Viewed by 205
Abstract
Tabanidae (horse flies and deer flies) are hematophagous insects that cause direct and indirect damage to animal production. The aims of this study were to determine the preferred site, time of day, and duration of tabanid feeding on beef cattle and identify factors [...] Read more.
Tabanidae (horse flies and deer flies) are hematophagous insects that cause direct and indirect damage to animal production. The aims of this study were to determine the preferred site, time of day, and duration of tabanid feeding on beef cattle and identify factors related to infestation by tabanids. The population of tabanids was surveyed on certain body parts of the beef cattle (fore udder, tail, navel, leg, dewlap, body, and under) during the morning hours (9.00–10.30 a.m.), midday (12.00–13.30 a.m.), and afternoon (15.30–17.00 p.m.) every day for 10 days. The findings showed that two genera, Tabanus Linnaeus, 1758, and Chrysops Meigen, 1803, landed on the cows. The leg was statistically significantly the most frequent landing site for tabanids (15.067 ± 7.54) compared with other parts. The average feeding duration for each insect was 2.76 ± 1.77 min. The results showed that a significant number of tabanids were present during midday, as compared with the morning and afternoon. Temperature was found to be positively associated with fly abundance. A regression model was derived in this study (y = 4.23x − 116.09). This information is important for tabanid control and prevention in beef cattle. Full article
(This article belongs to the Section Medical and Livestock Entomology)
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<p>The body parts of cattle where observations were focused: fore udder, tail, navel, leg (upper and lower), dewlap, body, and under.</p>
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<p>The frequency of feeding duration of tabanids on cattle.</p>
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16 pages, 874 KiB  
Article
Sustainable Energy Practices in Thailand and Japan: A Comparative Analysis
by Su Wutyi Hnin, Amna Javed, Jessada Karnjana, Chawalit Jeenanunta and Youji Kohda
Sustainability 2024, 16(16), 6877; https://doi.org/10.3390/su16166877 (registering DOI) - 10 Aug 2024
Viewed by 240
Abstract
This study investigates the comparative analysis of the divergent pathways of sustainable energy development in Thailand and Japan. It offers a nuanced analysis of their policy frameworks, technological advancements, and socioeconomic contexts. This study elucidates the distinct strategies of the two nations by [...] Read more.
This study investigates the comparative analysis of the divergent pathways of sustainable energy development in Thailand and Japan. It offers a nuanced analysis of their policy frameworks, technological advancements, and socioeconomic contexts. This study elucidates the distinct strategies of the two nations by leveraging a robust dataset from sources including the Electricity Generating Authority of Thailand (EGAT) and Japan’s Agency for Natural Resources and Energy (ANRE) toward renewable energy. The key findings indicate that Thailand has capitalized on policy instruments such as the Alternative Energy Development Plan 2018 (AEDP 2018) to augment its renewable energy capacity, particularly in the solar and biomass sectors. This policy-driven approach addresses the rural–urban energy divide and enhances energy access nationwide. Conversely, Japan’s trajectory is characterized by integrating technological innovations like smart grids and the Feed-in Tariff (FiT) system, which have catalyzed significant increases in solar energy adoption and efficiency. Japan places great emphasis on technological solutions that underscore its strategy to mitigate the legacy constraints of energy infrastructure post-Fukushima. The implications of these findings are extended beyond national borders, offering critical insights into the complex interplay between policy, technology, and social engagement in the renewable energy transition. This study highlights the potential for community-based renewable energy projects in Thailand to drive economic growth and social equity. At the same time, Japan’s experience illustrates the importance of regulatory reforms and technological leadership in overcoming structural barriers to energy innovation. These insights are particularly relevant for policymakers and stakeholders aiming to balance the imperatives of energy security, economic development, and environmental sustainability. Finally, this study emphasizes the need for tailored strategies that align renewable energy adoption with the unique contexts of each country, thereby enhancing global efforts against climate change. Full article
(This article belongs to the Section Energy Sustainability)
20 pages, 2047 KiB  
Article
Understanding Secondary Sarcopenia Development in Young Adults Using Pig Model with Chronic Pancreatitis
by Ewa Tomaszewska, Dorota Wojtysiak, Agnieszka Grzegorzewska, Małgorzata Świątkiewicz, Janine Donaldson, Marcin B. Arciszewski, Sławomir Dresler, Iwona Puzio, Sylwia Szymańczyk, Piotr Dobrowolski, Joanna Bonior, Maria Mielnik-Błaszczak, Damian Kuc and Siemowit Muszyński
Int. J. Mol. Sci. 2024, 25(16), 8735; https://doi.org/10.3390/ijms25168735 (registering DOI) - 10 Aug 2024
Viewed by 224
Abstract
Chronic pancreatitis (CP) in young individuals may lead to disease-related secondary sarcopenia (SSARC), characterized by muscle loss and systemic inflammation. In this study, CP was induced in young pigs, and serum levels of key hormones, muscle fiber diameters in various muscles, and the [...] Read more.
Chronic pancreatitis (CP) in young individuals may lead to disease-related secondary sarcopenia (SSARC), characterized by muscle loss and systemic inflammation. In this study, CP was induced in young pigs, and serum levels of key hormones, muscle fiber diameters in various muscles, and the mRNA expression of genes related to oxidative stress and programmed cell death were assessed. A decrease in muscle fiber diameters was observed in SSARC pigs, particularly in the longissimus and diaphragm muscles. Hormonal analysis revealed alterations in dehydroepiandrosterone, testosterone, oxytocin, myostatin, and cortisol levels, indicating a distinct hormonal response in SSARC pigs compared to controls. Oxytocin levels in SSARC pigs were significantly lower and myostatin levels higher. Additionally, changes in the expression of catalase (CAT), caspase 8 (CASP8), B-cell lymphoma 2 (BCL2), and BCL2-associated X protein (BAX) mRNA suggested a downregulation of oxidative stress response and apoptosis regulation. A reduced BAX/BCL2 ratio in SSARC pigs implied potential caspase-independent cell death pathways. The findings highlight the complex interplay between hormonal changes and muscle degradation in SSARC, underscoring the need for further research into the apoptotic and inflammatory pathways involved in muscle changes due to chronic organ inflammation in young individuals. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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<p>Muscle histomorphometry. (<b>A</b>) Representative images of the NADH-TR and MyHC stained sections of the diaphragm, the longissimus lumborum muscle, and the triceps brachii muscle in the control (C-pig) and cerulein-injected (SSARC-pig) groups, showing muscle fiber type variability. Type I fibers are stained brown, type IIa fibers are stained blue, and type IIb fibers are stained white; (<b>B</b>) Comparison of numbers of muscle fibers in the cross-section of the diaphragm, the longissimus lumborum muscle, and the triceps brachii muscle; (<b>C</b>) Comparison of the diameter of type I, IIa, and IIb muscle fibers in the diaphragm, longissimus lumborum muscle, and the triceps brachii muscle; (<b>D</b>) Comparison of the percentages of type I, IIa, and IIb muscle fibers in the diaphragm, longissimus lumborum muscle, and the triceps brachii muscle. In graphs, data are represented as the mean ± SE values (n = 5 in each group). Statistical significance: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 (two-tailed <span class="html-italic">t</span>-test adjusted for multiple comparisons using the Bonferroni method). In images in (<b>A</b>), all the scale bars show 100 μm.</p>
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<p>Time dependence of serum sarcopenic indicators during the experimental period: (<b>A</b>) dehydroepiandrosterone, DHEA; (<b>B</b>) testosterone, T; (<b>C</b>) cortisol; (<b>D</b>) oxytocin, OT; (<b>E</b>) myostatin, MNST; and (<b>F</b>) DHEA/cortisol ratio in pigs during the experimental period. Data are shown for the control group (C-pigs, black) and the cerulein-injected group (SSARC-pigs, red). The shaded area represents the 6-day-long period of daily cerulein injections (1 µg/kg b.w./day) in the SSARC-pig group. Data are presented as mean ± SE (n = 5 in each group). Statistical significance: * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001 (compared to day 0); # <span class="html-italic">p</span> &lt; 0.05; ## <span class="html-italic">p</span> &lt; 0.01 (between groups at a given time point).</p>
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<p>Relative gene expression of (<b>A</b>) antioxidant proteins, (<b>B</b>) signal proteins of programmed cell death, and (<b>C</b>) BAX/BCL2 mRNA ratio in pigs in the control (C-pig) and cerulein-injected (SSARC-pig) groups. The expression was normalized to the GAPDH housekeeping gene and is presented relative to the level observed in the control group. Due to the −ΔΔCT method’s exponential nature for calculating relative mRNA expression, the graphs depict the geometric means with SE (standard errors) (n = 5 in each group). Statistical significance: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 (two-tailed <span class="html-italic">t</span>-test adjusted for multiple comparisons using the Bonferroni method).</p>
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18 pages, 2701 KiB  
Article
Fungal Laccases and Fumonisin Decontamination in Co-Products of Bioethanol from Maize
by Marianela Bossa, Noelia Edith Monesterolo, María del Pilar Monge, Paloma Rhein, Sofía Noemí Chulze, María Silvina Alaniz-Zanon and María Laura Chiotta
Toxins 2024, 16(8), 350; https://doi.org/10.3390/toxins16080350 (registering DOI) - 10 Aug 2024
Viewed by 310
Abstract
Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. [...] Read more.
Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. The aims were to isolate and identify fungal laccase-producing strains, to evaluate laccase production, to determine the enzymatic stability under fermentation conditions, and to analyse the effectiveness in vitro of enzymatic extracts (EEs) containing laccases in degrading FB1. Strains belonging to Funalia trogii, Phellinus tuberculosus, Pleurotus ostreatus, Pycnoporus sanguineus and Trametes gallica species showed laccase activity. Different isoforms of laccases were detected depending on the evaluated species. For the FB1 decontamination assays, four enzymatic activities (5, 10, 15 and 20 U/mL) were tested, in the absence and presence of vanillic acid (VA) and 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) as redox mediators (1 and 10 mM). Trametes gallica B4-IMICO-RC EE was the most effective strain in buffer, achieving a 60% of FB1 reduction. Laccases included in EEs remained stable at different alcoholic degrees in maize steep liquor (MSL), but no significant FB1 reduction was observed under the conditions evaluated using MSL. This study demonstrate that although laccases could be good candidates for the development of a strategy to reduce FB1, further studies are necessary to optimise this process in MSL. Full article
(This article belongs to the Special Issue Effect of Mycotoxins on Crops and Their Prevention)
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<p>Specific and enzymatic activities of laccase enzymes from different enzymatic extracts of the laccase producing fungal isolates and the control strain (<span class="html-italic">Funalia trogii</span> B1-IMICO-RC), inoculated in broth for the laccase production. The enzymatic substrate used for the enzymatic activity determination was 2,2′-azino-di-3 ethylbenzothiazoline (ABTS). Error bars indicate the standard deviation within each treatment. Different capital letters denote significant differences between enzymatic activities (<span class="html-italic">p</span> value &lt; 0.01), and different lowercase letters indicate significant differences between specific activities (<span class="html-italic">p</span> value &lt; 0.01).</p>
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<p>Maximum specific activity observed during the 24-day incubation period of the laccase enzymes contained in the different enzymatic extracts of the laccase producing fungal isolates and the control strain (<span class="html-italic">Funalia trogii</span> B1-IMICO-RC), inoculated in broth for the laccase production. The enzymatic substrate used for the enzymatic activity determination was ABTS. Error bars indicate the standard deviation within each treatment. Different letters indicate significant differences between specific activities (<span class="html-italic">p</span> value &lt; 0.01).</p>
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<p>Native-PAGE (left side of each strain) vs. zymogram (right side of each strain) of the concentrated enzymatic extracts. Images of both gels were cut and presented one next to the other to show a better comparison of the results. Numbered arrows indicate the laccase or laccase isoenzyme synthesised by each strain. MW: molecular weight marker.</p>
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<p>Reduction percentages of FB<sub>1</sub> in the in vitro assays using buffer medium without redox mediators. A value of 100% signifies the total reduction of FB<sub>1</sub> compared to the control (0% reduction). Error bars represent the standard deviation of the replicates for each treatment. Different letters on the bars denote significant differences between treatments (<span class="html-italic">p</span> value &lt; 0.01).</p>
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<p>Stability of laccases from <span class="html-italic">Trametes gallica</span> B4-IMICO-RC (<b>a</b>) and <span class="html-italic">T. gallica</span> B7-IMICO-RC (<b>b</b>) under different alcoholic degrees in maize steep liquor (MSL) which was obtained from a bioethanol production process. M0, M12, M24 and M36 are samples collected at different times from the fermentation stage, corresponding to 0, 12, 24 and 36 h, respectively. The bars indicate the enzymatic activity (time 0 and 1 h) and the dotts linked by a dotted line represent the alcoholic degree of the MSL samples, measured at 0 h. Error bars indicate the standard deviation within each treatment. Different letters show significant differences between treatments (<span class="html-italic">p</span> value &lt; 0.01).</p>
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