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16 pages, 8084 KiB  
Article
Neurotropic Tick-Borne Flavivirus in Alpine Chamois (Rupicapra rupicapra rupicapra), Austria, 2017, Italy, 2023
by Norbert Nowotny, Maria Lucia Mandola, Isabella Monne, Zoltán Bagó, Chiara Nogarol, Alice Fusaro, Katharina Dimmel, Barbara Moroni, Lisa Guardone, Jolanta Kolodziejek, Elisa Palumbo, Gabriela Stanclova, Adi Steinrigl, Gabriele Fidler, Cristina Bertasio, Irene Bertoletti, Alessandro Bianchi, Mattia Calzolari, Paola Prati, Nadia Vicari, Angela Salomoni, Maria Francesca Priore, Federica Gobbo, Aitor Garcia-Vozmediano, Tom Loney, Ahmad Abou Tayoun, Alawi Alsheikh-Ali, Paola De Benedictis, Jeremy V. Camp, Zdenek Hubalek, Ivo Rudolf, Davide Lelli and Ana Morenoadd Show full author list remove Hide full author list
Viruses 2025, 17(1), 122; https://doi.org/10.3390/v17010122 - 16 Jan 2025
Abstract
The European subtype of tick-borne encephalitis virus (TBEV-Eur; species Orthoflavivirus encephalitidis, family Flaviviridae) was the only tick-borne flavivirus present in central Europe known to cause neurologic disease in humans and several animal species. Here, we report a tick-borne flavivirus isolated from [...] Read more.
The European subtype of tick-borne encephalitis virus (TBEV-Eur; species Orthoflavivirus encephalitidis, family Flaviviridae) was the only tick-borne flavivirus present in central Europe known to cause neurologic disease in humans and several animal species. Here, we report a tick-borne flavivirus isolated from Alpine chamois (Rupicapra rupicapra rupicapra) with encephalitis and attached ticks, present over a wide area in the Alps. Cases were detected in 2017 in Salzburg, Austria, and 2023 in Lombardy and Piedmont, Italy. The virus strains exhibit 94.8–97.3% nucleotide identities to each other and are more closely related to Louping ill viruses (LIV; Orthoflavivirus loupingi; 90–92% identities) than to TBEV-Eur (less than 88%). The chamois-derived virus strains, tentatively termed “Alpine chamois encephalitis virus”, form a well-supported independent genetic clade with Spanish goat encephalitis virus, clearly separated from other LIV. This supports its designation as a new virus subtype with the proposed shared taxonomic name “Spanish goat and Alpine chamois encephalitis virus subtype” within the species Orthoflavivirus loupingi. The zoonotic potential of this newly identified virus subtype as well as its host range in other animal species including farm animals needs to be further investigated. Full article
(This article belongs to the Special Issue Tick-Borne Viruses: Transmission and Surveillance, 2nd Edition)
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Figure 1
<p>Location of the three cases of Alpine chamois encephalitis virus infections in Austria and northern Italy, indicated by red dots. The straight line distances between the cases were 390 km [case 1 (Salzburg) and 2 (Piedmont)], 334 km [case 1 (Salzburg) and 3 (Lombardy)], and 66 km [case 2 (Piedmont) and 3 (Lombardy)], respectively.</p>
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<p>Histological sections from the brain of case 1. (<b>A</b>) Nonpurulent leptomeningitis: Slight lymphoplasmohistiocytic infiltrate (*); (<b>B</b>–<b>D</b>) Nonpurulent encephalitis; (<b>B</b>): Slight lymphoplasmohistiocytic perivascular infiltrate (*); (<b>C</b>) Two glial nodules in brain stem (*); (<b>D</b>) Glial shrubbery in the cerebellum (*). Microphoto, H&amp;E, bar (<b>A</b>) = 100 µm, bars (<b>B</b>–<b>D</b>) = 180 µm.</p>
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<p>Maximum likelihood phylogenetic tree of the complete genomes of Alpine chamois encephalitis viruses and other TBEV, LIV, and LIV-like sequences constructed using the IQtree software and the GTR + F+I + G4 model according to BIC and a bootstrap of 1000 replicates. Sequences investigated in this study are shown in blue. The sequences of Spanish sheep encephalitis virus, Greek goat encephalitis virus, and Turkish sheep encephalitis virus are highlighted in yellow, and the sequence of Spanish goat encephalitis virus is highlighted in turquoise.</p>
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<p>SSE similarity plots over the whole genome sequences between the Alpine chamois encephalitis viruses (ACEV) from Austria and Italy, the Spanish goat encephalitis virus (SGEV), and the reference strains of tick-borne encephalitis virus European subtype (TBEV-Eur) and Louping ill virus (LIV). Tick and chamois virus sequences for Piedmont and Lombardy are tagged as a group. Similarity plots were generated using SSE version 1.2 using a sliding window of 600 and a step size of 100 nucleotides. The percentage of identities (<span class="html-italic">y</span>-axis) between 50 and 100% is shown over the whole genome alignment (<span class="html-italic">x</span>-axis).</p>
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<p>Structural alignment of the E protein. The secondary structure elements are defined based on an ESPript algorithm and are labeled as in a previous report on the LIV structure (PDB no. 6J5C) and are presented on top (Spiral lines indicate helices, arrows represent β strands and TT letters turns). The Arabic numerals 1–6 indicate cysteine residues that pair to form disulfide bonds. Lines above the sequences indicate domains I (red), II (yellow), and III (blue), as well as the fusion peptide loop (pink) and the dI/dIII and dIII/stem linkers (purple). Austrian and Italian ACEV strains are shown in a box.</p>
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<p>Map of Europe. Location of the three cases of Alpine chamois encephalitis virus infections in Austria and Northern Italy and approximate location of the cases of Spanish goat encephalitis virus infections in the Principality of Asturias (capital: Oviedo) in Northwestern Spain are displayed. The approximate straight line distance between cases of Alpine chamois encephalitis virus and Spanish goat encephalitis virus infections (for the latter, the exact location was not published) is between 1170 km (Piedmont case, Oviedo) and 1550 km (Salzburg case, Oviedo). The base map shapefile was obtained from the Geographic Information System of the EU Commission via the Eurostat webpage—the 2021 version at 20 M resolution showing NUTS level 3 regions—and annotated in QGIS v.3.4.15.</p>
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13 pages, 932 KiB  
Article
The Anthelmintic Activity of Nepeta racemosa Lam. Against Gastrointestinal Nematodes of Sheep: Rosmarinic Acid Quantification and In Silico Tubulin-Binding Studies
by Büşra Karpuz Ağören, Mahmut Sinan Erez, Esma Kozan, Aydın Dağyaran, Mevlüt Akdağ, Eduardo Sobarzo-Sánchez and Esra Küpeli Akkol
Pathogens 2025, 14(1), 77; https://doi.org/10.3390/pathogens14010077 - 15 Jan 2025
Viewed by 513
Abstract
Gastrointestinal nematodes (GINs) inflict significant economic losses on sheep and goat farming globally due to reduced productivity and the development of anthelmintic resistance. Sustainable control strategies are urgently needed including the exploration of medicinal plants as safer alternatives to chemical anthelmintics. This genus [...] Read more.
Gastrointestinal nematodes (GINs) inflict significant economic losses on sheep and goat farming globally due to reduced productivity and the development of anthelmintic resistance. Sustainable control strategies are urgently needed including the exploration of medicinal plants as safer alternatives to chemical anthelmintics. This genus of plants is used for anti-inflammatory, antioxidant, and antimicrobial activities. In this study, we aimed to evaluate the anthelmintic activities of Nepeta racemosa Lam. MeOH extract, n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), n-buthanol (n-BuOH) and aqueous (H2O) subextracts, and quantify rosmarinic acid in the active extract by the HPLC method, and perform in silico molecular docking studies of rosmarinic acid to examine its binding interactions with tubulin. The anthelmintic activity of the plant extracts on gastrointestinal nematode eggs and larvae (L3) of the sheep was assessed using in vitro test methods such as the egg hatch assay and larval motility assay, conducted over a 24 h period (1, 2, 3, 4, 6, 8, 24). All extracts exhibited 100% effectiveness in the egg hatch inhibition assay, regardless of concentration (50–1.5625 mg/mL). The EtOAc subextract shows the highest effectiveness at 79.66%, followed by the MeOH extract at 74.00%, water at 64.00%, n-hexane at 67.00%, and DCM at 61.00%, and the lowest effectiveness is observed with n-BuOH at 51.66% in the larval motility assay. The major compound of EtOAc extract, the most active extract of N. racemosa, was determined as rosmarinic acid and its amount in the extract was determined as 14.50 mg/100 mg dry extract. The amount of rosmarinic acid in the MeOH extract was found to be 0.21 mg/100 mg dry extract. n-Hexane, DCM, n-BuOH, and H2O extracts’ rosmarinic acid content was lower than the LOQ value. As tubulin plays an important role in the mechanism of anthelmintics, the major compound of the most active extract (NR-EtOAc) rosmarinic acid was docked onto the colchicine-binding site of the tubulin (5OV7) protein. Rosmarinic acid showed a similar activity spectrum to the anthelmintic drug albendazole. The discovery of low-cost and low-toxicity anthelmintic compounds is very important. Full article
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Figure 1
<p>The HPLC chromatograms of the MeOH extract, EtOAc subextract, and rosmarinic acid (330 nm).</p>
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<p>In silico binding interactions between rosmarinic acid and tubulin (PDB code: 5OV7). (<b>A</b>) Rosmarinic acid docked on the colchicine-binding site. (<b>B</b>) Three-dimensional representation of interactions between rosmarinic acid and colchicine-binding-site residues. (<b>C</b>) Two-dimensional representation of interactions between rosmarinic acid and colchicine-binding-site residues. Green, purple, pink, and yellow dashed lines represent hydrogen bonds, pi–sigma bonds, pi–alkyl bonds, and pi–sulphur bonds, respectively.</p>
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15 pages, 1407 KiB  
Article
Efficacy of Fat Supplements with Different Unsaturated/Saturated FA Ratios Undergoing First Postpartum Ovulation in Lactating Anovulatory Goats
by Caroline P. Silva, César C. L. Fernandes, Juliana P. M. Alves, Camila M. Cavalcanti, Felipe B. B. Oliveira, Alfredo J. H. Conde, Diana Celia S. N. Pinheiro, Darcio I. A. Teixeira, Anibal C. Rego and Davide Rondina
Vet. Sci. 2025, 12(1), 60; https://doi.org/10.3390/vetsci12010060 - 15 Jan 2025
Viewed by 232
Abstract
We investigated whether microalgae or linseed supply during the early postpartum period affects ovarian restimulation and supports the first postpartum ovulation in lactating anovulatory goats. Thirty-eight An-glo-Nubian-crossbred adult goats were allocated into three groups, one with a control diet (n = 12), [...] Read more.
We investigated whether microalgae or linseed supply during the early postpartum period affects ovarian restimulation and supports the first postpartum ovulation in lactating anovulatory goats. Thirty-eight An-glo-Nubian-crossbred adult goats were allocated into three groups, one with a control diet (n = 12), fed a total mixed ration (TMR) comprising chopped elephant grass and concentrate; an algal diet (n = 13), fed TMR + green microalgae (1% dry matter); and a linseed diet (n = 13), TMR + linseed (12% dry matter). Supplements were furnished from the second to fifth week (time of weaning). Goats were estrus synchronized on day 40 by insertion of an intravaginal CIDR device for 5 days, after which 0.075 mg PGF2α was applied to induce ovulation, and estrus was monitored for 72 h. From the 5th–15th day of ovulation induction, the corpus luteum (CL) area and progesterone rate were monitored. The algal and linseed groups showed lower feed intake (p < 0.001) and higher (p < 0.001) triglyceride levels/follicle numbers, respectively. After estrus induction, no differences were observed in estrus response; however, the linseed group showed more and larger growing follicles (p = 0.016 and p < 0.01), a higher ovulation rate (p < 0.05), a larger CL area (p < 0.05), and higher progesterone levels (p < 0.001). Linseed after delivery stimulates follicular growth before and after ovulation induction, favoring better CL quality during the first ovulation. Full article
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Graphical abstract

Graphical abstract
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<p>Timeline for the experimental design used in the study, describing the period of dietary supplementation and the hormonal protocol for estrus synchronization.</p>
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<p>Kidney fat thickness (<b>A</b>), subcutaneous loin fat thickness (<b>B</b>), triglyceride levels (<b>C</b>), and body weight (<b>D</b>), with measurements performed after delivery and during nutritional supplementation from 2 weeks to 5 weeks after delivery (21 days) in goats. Data are plotted as mean ± SEM. The <span class="html-italic">p</span>-value for the ANOVA effects for supplementation period are shown in the figure. Time, ANOVA effect for interval of assessment used. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, differences between groups.</p>
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<p>Total follicles counted by ultrasonography performed after delivery and during nutritional supplementation from 2 weeks to 5 weeks after delivery (21 days) in goats. Data are plotted as mean ± SEM. The <span class="html-italic">p</span>-value for the ANOVA effects for the supplementation period are shown in the figure. Time, ANOVA effect for interval of assessment used. * <span class="html-italic">p</span> &lt; 0.05 differences between groups.</p>
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<p>Peripheral progesterone levels (<b>A</b>) and luteal area (<b>B</b>), measured after ovulation induction by hormonal treatment started 40 days after delivery in goats previously supplemented with algae or linseed. Data are plotted as mean ± SEM. (<b>A</b>) The <span class="html-italic">p</span>-value for the ANOVA effects of diet group; the interval of assessments and interaction are shown in the figure. a,b <span class="html-italic">p</span> &lt; 0.05 differences between nutritional groups, shown in (<b>B</b>). * <span class="html-italic">p</span> &lt; 0.05, differences between groups.</p>
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12 pages, 1974 KiB  
Article
Occurrence and Multi-Locus Genotyping of Giardia duodenalis in Black Goats from Fujian Province, China
by Shou-Xiao Huang, Kai Hu, Peng-Fei Fu, Si-Ang Li, Yang Liu, Zhipeng Niu and Dong-Hui Zhou
Animals 2025, 15(2), 199; https://doi.org/10.3390/ani15020199 - 13 Jan 2025
Viewed by 354
Abstract
Giardia duodenalis is a zoonotic parasite that causes gastrointestinal diseases in both humans and animals. To evaluate the prevalence and genetic diversity of G. duodenalis in black goats, we collected 539 fecal samples from nine districts in Fujian Province, China. The presence of [...] Read more.
Giardia duodenalis is a zoonotic parasite that causes gastrointestinal diseases in both humans and animals. To evaluate the prevalence and genetic diversity of G. duodenalis in black goats, we collected 539 fecal samples from nine districts in Fujian Province, China. The presence of G. duodenalis was confirmed through nested PCR targeting the SSU rRNA gene, and genotyping was performed at the beta-giardin, glutamate dehydrogenase, and triosephosphate isomerase loci. Among the samples, 115 tested positive, yielding an overall infection rate of 21.34%. Assemblages A and E were identified, with assemblage E being predominant. Statistical analysis revealed significant regional differences in infection rates (p < 0.01), with Zhangzhou exhibiting the highest infection rate (39%) and Fuzhou the lowest (3.13%). No significant differences in infection rates were observed based on age: 24.56% (56/228) for goats <1 year, 14.92% (27/181) for goats 1–2 years, 26.8% (26/97) for goats 2–3 years, and 18.18% (6/33) for goats ≥ 3 years. Similarly, no significant differences were found between sexes: 24.84% (40/161) for males and 19.84% (75/378) for females. Notably, assemblage A, a zoonotic genotype, was detected, indicating a potential risk of cross-species transmission. This study contributes to a deeper understanding of G. duodenalis in black goats and provides critical data for the development of targeted control strategies in Fujian Province. Full article
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<p>Distribution of sampling sites in Fujian.</p>
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<p>The phylogenetic tree illustrating the evolutionary relationships of <span class="html-italic">G. duodenalis</span> from sheep was constructed based on SSU rRNA gene sequences using maximum likelihood analysis. Sequences representative of each sequence type identified in this study were included in the phylogenetic analysis. The arrow indicates the genotypes identified in this experiment.</p>
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<p>Phylogenetic evolutionary tree diagram of the <span class="html-italic">bg</span> (<b>A</b>), <span class="html-italic">gdh</span> (<b>B</b>), and <span class="html-italic">tpi</span> (<b>C</b>) loci. The arrow indicates the genotypes identified in this experiment.</p>
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14 pages, 1707 KiB  
Article
Ehrlichia Species in Dromedary Camels (Camelus dromedarius) and Ruminants from Somalia
by Aamir M. Osman, Ahmed A. Hassan-Kadle, Marcos R. André, Flávia C. M. Collere, Amir Salvador Alabí Córdova, Fabiano Montiani-Ferreira, Thállitha S. W. J. Vieira, Abdalla M. Ibrahim, Abdulkarim A. Yusuf, Rosangela Z. Machado and Rafael F. C. Vieira
Pathogens 2025, 14(1), 65; https://doi.org/10.3390/pathogens14010065 - 13 Jan 2025
Viewed by 386
Abstract
Ehrlichioses, caused by Ehrlichia species, are tick-borne diseases (TBDs) that affect animals and humans worldwide. This study aimed to investigate the molecular occurrence of Ehrlichia spp. in 530 animals (155 Dromedary camels, 199 goats, 131 cattle, and 45 sheep) in the Benadir and [...] Read more.
Ehrlichioses, caused by Ehrlichia species, are tick-borne diseases (TBDs) that affect animals and humans worldwide. This study aimed to investigate the molecular occurrence of Ehrlichia spp. in 530 animals (155 Dromedary camels, 199 goats, 131 cattle, and 45 sheep) in the Benadir and Lower Shabelle regions of Somalia. Blood DNA samples were tested for PCR targeting dsb and sodB genes of Ehrlichia spp. and PCS20 and map1 genes of E. ruminantium. The obtained sequences were submitted for phylogenetic analyses. Ehrlichia spp. were detected in 26.4% (140/530) of animals by dsb-PCR, with the highest prevalence in dromedary camels (54.8%), followed by cattle (29.8%), goats (7.0%), and sheep (4.4%). Dromedary camels, cattle, and goats had significantly higher infection odds compared to sheep (p < 0.05). Among dsb-PCR-positive samples, 76.9% (30/39) of cattle tested sodB-positive, while other species were negative. E. ruminantium was detected in 13.7% (18/131) of cattle by pCS20-PCR, but none were positive for the map1 gene. Phylogenetic analysis confirmed E. minasensis in camels, sheep, and goats and E. ruminantium in cattle, marking the first molecular evidence of E. minasensis in dromedary camels, sheep, and goats globally, and E. ruminantium in cattle from Somalia. These findings emphasize the need for further research on its economic and public health impact. Full article
(This article belongs to the Section Bacterial Pathogens)
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<p>Phylogenetic tree inferred by using maximum likelihood inference and GTR+G evolutionary model based on an alignment of 390 bp of the <span class="html-italic">dsb</span> gene. The sequences detected are highlighted in red in the present study. <span class="html-italic">Ehrilichia muris</span> was used as the outgroup.</p>
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<p>Phylogenetic tree inferred by using maximum likelihood inference and GTR+G evolutionary model based on an alignment of 300 bp of the <span class="html-italic">sodB</span> gene. The sequences detected are highlighted in red in the present study. The numbers at the nodes correspond to posterior probability values higher than 50% accessed with 1000 replicates. <span class="html-italic">A. marginale</span> was used as outgroups.</p>
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<p>Phylogenetic tree inferred by using maximum likelihood inference and GTR+G evolutionary model based on an alignment of 280 bp of the <span class="html-italic">E. ruminantium PCS20</span> gene. The sequences detected are highlighted in red in the present study. The numbers at the nodes correspond to posterior probability values higher than 50% accessed with 1000 replicates. <span class="html-italic">E. canis</span> was used as the outgroup.</p>
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14 pages, 4880 KiB  
Article
Effect of Liquid Marble 3D Culture System on In Vitro Maturation and Embryo Development of Prepubertal Goat Oocytes
by Andrea Podda, Linda Dujíčková, Federica Ariu, Giovanni Giuseppe Leoni, Dolors Izquierdo, Maria-Teresa Paramio and Luisa Bogliolo
Animals 2025, 15(2), 188; https://doi.org/10.3390/ani15020188 - 12 Jan 2025
Viewed by 360
Abstract
Suboptimal culture conditions during in vitro maturation (IVM) affect oocyte developmental competence and the viability of the resulting embryo. Three-dimensional (3D) culture systems provide a more biologically appropriate environment compared to traditional two-dimensional (2D) cultures. The aim of this study was to evaluate [...] Read more.
Suboptimal culture conditions during in vitro maturation (IVM) affect oocyte developmental competence and the viability of the resulting embryo. Three-dimensional (3D) culture systems provide a more biologically appropriate environment compared to traditional two-dimensional (2D) cultures. The aim of this study was to evaluate the effect of liquid marble (LM) microbioreactors as a 3D culture system on IVM and the subsequent embryo development of prepubertal goat oocytes. The cumulus–oocyte complexes (COCs) recovered from prepubertal goat ovaries underwent IVM in drops under oil (the 2D system and the control group) and in the 3D LM system (the LM group). After IVM, oocytes were parthenogenetically activated and cultured until the blastocyst stage. The control and LM groups showed similar rates of nuclear maturation (52.17% and 44.12%) and blastocyst formation (10.64% and 10.10%). Reactive oxygen species and glutathione levels and the density of transzonal projections (TZPs) in oocytes did not differ between groups. The LM system increased mitochondrial activity and modified the organization of these organelles in the oocyte cytoplasm compared to the control group. The LM microbioreactor demonstrated the ability to improve the mitochondrial status of the oocytes and was not harmful for oocyte IVM and subsequent embryo development. Therefore, LM could be used as a 3D cost-effective culture system for the IVM of prepubertal goat oocytes. Full article
(This article belongs to the Section Animal Reproduction)
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<p>Schematic picture of IVM culture systems. (<b>A</b>) 2D system with three drops of maturation medium containing oocytes covered with mineral oil. (<b>B</b>) The LM system—three 35 mm Petri dishes with LM droplets containing oocytes, placed in a 90 mm Petri dish with sterile water, to ensure enough humidity during the culture. (<b>C</b>–<b>G</b>) Preparation of LM droplets. (<b>C</b>) A drop of maturation media containing oocytes is being aspirated with a pipette. (<b>D</b>) The drop is slowly released into the dish with the powder. (<b>E</b>) After release, gently rolling of the drop in the powder is required to fully cover the drop. (<b>F</b>) A well-coated drop of maturation medium with the hydrophobic powder in the Petri dish. (<b>G</b>) The drop is aspirated by a Pasteur pipette and placed into the 35 mm Petri dish.</p>
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<p>Effect of the LM system on the meiotic maturation of prepubertal goat oocytes. GV: germinal vesicle, GVBD: germinal vesicle breakdown, MI: metaphase I, MII: metaphase II. Different superscripts indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the LM system on the TZPs density of prepubertal goat oocytes. (<b>A</b>) Phalloidin-FITC average fluorescence intensity in the zona area of COCs after recovery (T0), 6 h (T6) and 24 h (T24) of IVM. (<b>B</b>) Representative confocal images of oocytes with stained TZPs (actin, green) which appear as continuous filaments going from the CCs to the oocyte through the zona region. Values are expressed as the mean ± SEM. Different superscripts indicate significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the LM system on (<b>A</b>) the intracellular ROS and GSH levels of prepubertal goat oocytes based on the relative fluorescence intensity. (<b>B</b>) Epifluorescence photomicrographs of oocytes stained with H<sub>2</sub>DCF-DA and CellTracker Blue to detect ROS (green) and GSH (blue) levels respectively, in the tested groups. Values are expressed as the mean ± SEM.</p>
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<p>(<b>A</b>) Effect of the LM system on mitochondrial activity in prepubertal goat oocytes. (<b>B</b>) Representative images of mitochondrial activity in oocytes. Values are expressed as the mean ± SEM. Different superscripts indicate significant differences at <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Representative images of distribution patterns of mitochondria (red signal) in goat oocytes: (<b>A</b>) diffused, (<b>B</b>) semi-peripheral and (<b>C</b>) peripheral.</p>
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<p>Effect of the LM system on embryo development. (<b>A</b>) Cleavage at 24 and 30 h post activation, and the blastocyst rate after 8 days from activation. (<b>B</b>) Proportion of normal, expanded, hatching and hatched blastocysts on day 8 of culture in the tested groups. Differences between groups were not significant.</p>
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20 pages, 2018 KiB  
Article
A Comprehensive Analysis of Echinococcus granulosus Infections in Children and Adolescents: Results of a 7-Year Retrospective Study and Literature Review
by Cristina Maria Mihai, Ancuta Lupu, Tatiana Chisnoiu, Adriana Luminita Balasa, Ginel Baciu, Vasile Valeriu Lupu, Violeta Popovici, Felicia Suciu, Florin-Daniel Enache, Simona Claudia Cambrea and Ramona Mihaela Stoicescu
Pathogens 2025, 14(1), 53; https://doi.org/10.3390/pathogens14010053 - 10 Jan 2025
Viewed by 567
Abstract
Cystic echinococcosis (CE) is a neglected tropical parasitic disease linked with significant social and economic burdens worldwide. The scientific community has minimal information on echinococcosis in Romanian people, and hospital medical records are the only sources that may be used to investigate its [...] Read more.
Cystic echinococcosis (CE) is a neglected tropical parasitic disease linked with significant social and economic burdens worldwide. The scientific community has minimal information on echinococcosis in Romanian people, and hospital medical records are the only sources that may be used to investigate its status. A 7-year retrospective clinical study on pediatric patients with CE from Southeast Romania was performed, and 39 children and adolescents were included, aged 2–15 years old. They were hospitalized with cystic echinococcosis in the Pediatric Department and Pediatric Surgery Department of Constanta County Clinical Emergency Hospital “St. Apostle Andrew” between 1 January 2017 and 1 October 2024. Twenty-nine (74.36%) pediatric patients came from rural zones, and 10 (25.64%) had urban residences. In total, 28 children (71.79%) had contact with four different animals (dogs, goats, pigs, and sheep); only four were from urban zones, and they had contact only with dogs. Data regarding the length of hospital stay, cyst location, and complications were collected and analyzed. According to the medical files, the diagnosis was established using imaging techniques and serological tests for CE. IgE and IgG reported appreciable variations in correlation with all parameters, and significant differences (p < 0.05) were recorded. IgE levels considerably increased in cases of no animal contact, pulmonary involvement, complications, surgical treatment, and multiple hospitalizations. Moderate IgE values were recorded in cases of urban residences, pig and sheep contact, and hepatic involvement. The IgG concentration considerably increased with sheep contact and moderately increased in cases of rural zones, hepatic involvement, complications, and surgical treatment. The results show that incidental discovery, symptoms, complications, multiple dissemination, pulmonary involvement, and dog and pig contact increase the hospitalization time. Extensive data analysis supports our results. Our findings highlight the complexity of managing E. granulosus infections in children and evidence the importance of a multidisciplinary approach, combining early diagnostic tools, tailored medical therapy, and careful surgical intervention when necessary. Full article
(This article belongs to the Special Issue One Health: New Approaches, Research and Innovation to Zoonoses)
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Figure 1
<p>The main symptoms associated with organs involved in CE in pediatric patients. AP = abdominal pain; AP +1 = abdominal pain +1 = abdominal pain with fever; AP +2 = abdominal pain +2 = abdominal pain + fever + headache and abdominal pain + nausea and vomiting; Cough +1 = cough associated with one of the following symptoms: fever, hemoptysis, shortness of breath, or thoracic back pain; Cough +2 = cough + fever and hemoptysis, cough + fever and thoracic pain, cough + shortness of breath and fever; Other 1 = axillary adenopathy, back pain, left thoracic pain, tachycardia, or vomiting; ID = incidental discovery; MD = multiple organ diffusion; C = complications; HD = hospitalization days.</p>
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<p>(<b>A</b>) The main symptoms associated with MD. (<b>B</b>) The correlation between MD and CE localization. (<b>C</b>) MD association with complications. (<b>D</b>) Complication types. AP = abdominal pain; AP +1 = abdominal pain +1 = abdominal pain with fever; AP +2 = abdominal pain +2 = abdominal pain + fever + headache and abdominal pain + nausea and vomiting; Cough +1 = cough associated with one of the following symptoms: fever, hemoptysis, shortness of breath, or thoracic back pain; Cough +2 = cough + fever and hemoptysis, cough + fever and thoracic pain, cough + shortness of breath and fever; Other 1 = axillary adenopathy, back pain, left thoracic pain, tachycardia, or vomiting; ID = incidental discovery; MD = multiple organ diffusion; C = complications; HD = hospitalization days.</p>
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<p>Correlations between the main symptoms at presentation, organ involvement complications, and hospitalization days. AP = abdominal pain; AP +1 = abdominal pain +1 = abdominal pain with fever; AP +2 = abdominal pain +2 = abdominal pain + fever and headache and abdominal pain + nausea and vomiting; Cough +1 = cough associated with one of the following symptoms: fever, hemoptysis, shortness of breath, or thoracic back pain; Cough +2 = cough + fever and hemoptysis, cough + fever and thoracic pain, cough + shortness of breath and fever; Other 1 = axillary adenopathy, back pain, left thoracic pain, tachycardia, or vomiting; ID = incidental discovery; MD = multiple organ diffusion; C = complications.</p>
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<p>Correlations between organ involvement, laboratory analyses, treatment type, and hospitalization days. MH = multiple hospitalizations; MD = multiple organ diffusion; TM = medication (with albendazole); TS = surgical treatment; Eos = eosinophilia (N/µL); IgE (i.u./mL); IgG (i.u./mL).</p>
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18 pages, 3466 KiB  
Article
Isolation of a Novel Caprine Eimeria christenseni Strain (GC) in Canary Islands and Analysis of Parasitological, Clinical, and Pathological Findings on Experimentally Infected Goat Kids
by Emilio Barba, José Manuel Molina, Francisco Rodríguez, Otilia Ferrer, María Carmen Muñoz, Liliana M. R. Silva, María Cristina Del Río, José Adrián Molina, Anja Taubert, Carlos Hermosilla and Antonio Ruiz
Animals 2025, 15(2), 139; https://doi.org/10.3390/ani15020139 - 8 Jan 2025
Viewed by 393
Abstract
Eimeria christenseni is considered among the most pathogenic Eimeria species in goats. The aim of this study was to isolate an E. christenseni strain and to assess its infectivity, pathogenicity, and ability to develop a protective immune response. After previous collection of E. [...] Read more.
Eimeria christenseni is considered among the most pathogenic Eimeria species in goats. The aim of this study was to isolate an E. christenseni strain and to assess its infectivity, pathogenicity, and ability to develop a protective immune response. After previous collection of E. christenseni-positive faeces, purification of oocysts, and amplification in donor animals, an experimental infection was carried out. A total of 19 kids were divided into three groups: primary-infected and challenged, challenge control, and uninfected control. Infections were performed orally with 2 × 105 sporulated oocysts per animal. Oocyst shedding, clinical signs, and production parameters, in addition to haematological and histopathological features, were monitored. The results showed that the Gran Canaria (GC) E. christenseni strain had similar morphological and biological characteristics to those previously described, but no significant clinical signs were observed despite the high oocyst counts here recorded. The novel strain isolated would therefore be of low pathogenicity but still able to develop significant immunoprotective responses upon challenge infections. Its biological similarities to highly pathogenic species such as Eimeria ninakohlyakimovae and Eimeria arloingi might enable comparative studies aimed at developing alternative strategies for drug treatments, including Eimeria species (strain)-specific vaccination strategies for the efficient control of goat coccidiosis. Full article
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<p>Morphological characteristics of oocysts from the <span class="html-italic">E. christenseni</span> GC strain. (<b>A</b>) Full sporulated oocyst. (<b>B</b>) Sporulated oocyst with semidetached micropillar cap. (<b>C</b>) Sporulated oocyst with completely detached micropillar cap.</p>
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<p>OPG counts in goat kids challenge-infected (G1) and primary-infected (G2) with <span class="html-italic">E. christenseni</span> GC strain. The arrow shows the time of challenge infection (G1) or primary infection (G2). Data are expressed as mean ± SEM. (*) <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Weekly variation in body weight in goat kids challenge-infected (G1) and primary-infected (G2) with <span class="html-italic">E. christenseni</span> GC strain compared to uninfected controls (G3). Data are expressed as mean ± SEM. (**) <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Main histopathological findings in the intestinal mucosa of goat kids experimentally infected with <span class="html-italic">E. christenseni</span>. (<b>1</b>) Prominent hyperaemia and interstitial oedema of the lamina propria associated with mononuclear infiltration. (<b>2</b>) Intense diffuse mononuclear infiltration in the lamina propria. (<b>3</b>–<b>5</b>) Numerous intracellular macro- and microgametocytes at different stages of development. (<b>6</b>) Intense infiltration of inflammatory cells, rich in leucocytes eosinophils, associated with intracellular gamonts, schizonts, and oocysts in the lumen of necrotic glands.</p>
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21 pages, 891 KiB  
Review
Leveraging Microalgae to Achieve Zero Hunger: Enhancing Livestock Feed for Nutritional Security
by Leila Urrutia-Mazzuca, Marcia Mazzuca, María José Ibáñez-González and Tania Mazzuca-Sobczuk
Biomass 2025, 5(1), 4; https://doi.org/10.3390/biomass5010004 - 8 Jan 2025
Viewed by 403
Abstract
Achieving “Zero Hunger” (SDG 2) requires overcoming complex challenges, especially in vulnerable communities in developing countries. Livestock plays a key role in food security, but limited resources threaten productivity, prompting interest in innovative solutions like microalgae supplementation in ruminant diets. Microalgae offer potential [...] Read more.
Achieving “Zero Hunger” (SDG 2) requires overcoming complex challenges, especially in vulnerable communities in developing countries. Livestock plays a key role in food security, but limited resources threaten productivity, prompting interest in innovative solutions like microalgae supplementation in ruminant diets. Microalgae offer potential benefits by enhancing productivity and nutrition while addressing local protein deficiencies. However, barriers such as economic costs, processing requirements, and resistance to changing traditional feeding practices present challenges. This review examines the feasibility of microalgae-based livestock feed as a sustainable strategy to improve food security, particularly in arid, climate-affected regions. Biomass yield estimates suggest that small-scale cultivation can meet livestock nutritional needs; for example, a 22-goat herd would require approximately 88 g of microalgae per day to enrich meat with polyunsaturated fatty acids. Semi-continuous production systems could enable smallholders to cultivate adequate biomass, using local agricultural resources efficiently. This approach supports food security, improves meat quality, and strengthens community resilience. Collaboration among researchers, extension services, and local farmers is essential to ensure the effective adoption of microalgae feed systems, contributing to a sustainable future for livestock production in vulnerable regions. Full article
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<p>Differences between batch (<b>left</b>) and continuous (<b>right</b>) operation modes for a bubble column to produce microalgae. On top: schematic representation of the column operation; (<b>a</b>) working in batch between the operations of inoculation and the final harvesting, and (<b>b</b>) working in continuous after inoculation. Bottom: feasible (ideal) evolution of biomass concentration during the same time (<b>c</b>) after the first cycle of inoculation and up to the harvesting of the second cycle (total time = 57 days) (<b>d</b>) after the first inoculation on day 0 and during the 57 days of continuous culture. Continuous culture is established at day 13.</p>
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<p>Experience in cultivating microalgae on a family scale with secondary school students. Reprinted with permission of IQ-boletin. Figures (<b>a</b>–<b>d</b>) are home-made bioreactors for culturing <span class="html-italic">Spirulina</span>; Figure (<b>e</b>) shows the harvesting systems designed by secondary school students and (<b>f</b>) shows the drying system selected by the students.</p>
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14 pages, 2325 KiB  
Article
Genome-Wide Scans for Selection Signatures in Haimen Goats Reveal Candidate Genes Associated with Growth Traits
by Zhen Zhang, Jiafeng Lu, Yifei Wang, Zhipeng Liu, Dongxu Li, Kaiping Deng, Guomin Zhang, Bingru Zhao, Peihua You, Yixuan Fan, Feng Wang and Ziyu Wang
Biology 2025, 14(1), 40; https://doi.org/10.3390/biology14010040 - 7 Jan 2025
Viewed by 428
Abstract
Understanding the genetic characteristics of indigenous goat breeds is vital for their conservation and breeding. Haimen goats, native to China’s Yangtze River Delta, possess distinctive traits such as white hair, moderate growth rate, high-quality meat, and small body size. However, knowledge regarding the [...] Read more.
Understanding the genetic characteristics of indigenous goat breeds is vital for their conservation and breeding. Haimen goats, native to China’s Yangtze River Delta, possess distinctive traits such as white hair, moderate growth rate, high-quality meat, and small body size. However, knowledge regarding the genetic structure and germplasm characteristics of Haimen goats remains limited. In this study, we performed 20× whole-genome resequencing of 90 goats (60 Haimen goats and 30 Boer goats) to identify single-nucleotide polymorphisms (SNPs) and insertions/deletions (Indels) associated with growth traits. Here, we analyzed population genetic structure and genome-wide selection signatures between the Haimen and Boer goats based on whole-genome resequencing data. The principal component analysis (PCA) and neighbor-joining (N-J) tree results demonstrated significant genetic differentiation between the Haimen and Boer goats. The nucleotide diversity (Pi) and linkage disequilibrium (LD) decay results indicated higher genomic diversity in the Haimen goat population. Furthermore, selective sweep analysis identified candidate genes associated with growth traits. These genes exhibited strong selection signatures and were related to body size (DONSON, BMPR1B, and EPHA5), muscle development (GART, VGLL3, MYH15), and fat metabolism (ADAMTS5, LRP6, XDH, CPT1A, and GPD1). We also identified growth-related candidate genes (NCOR1, DPP6, NOTCH2, and FGGY) specific to Haimen goats. Among these genes, pancreatic lipase-related protein 1 (PNLIPRP1) emerged as the primary candidate gene influencing growth phenotypes. Further analysis revealed that a 26 bp Indel in PNLIPRP1 increased its gene expression, suggesting that this Indel could serve as a molecular marker for early marker-assisted selection, potentially enhancing early growth in goats. These findings provide valuable molecular markers and candidate genes for improving growth traits in Haimen goat breeding. Full article
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<p>Population genetics analyses and genome-wide scanning. (<b>A</b>,<b>B</b>) The distribution of SNPs and Indels on autosomal chromosomes of goats. (<b>C</b>) Representative photograph of Boer and Haimen goat (HM). (<b>D</b>) Principal component plot. The first (PC1) and second (PC2) principal components are displayed. (<b>E</b>) Neighbor-joining phylogenetic tree of 30 Boer and 60 HM. (<b>F</b>) Distribution of nucleotide diversity (Pi) in Boer and HM. (<b>G</b>) LD decay in two species.</p>
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<p>Functional enrichment analysis of candidate genes. (<b>A</b>) GO term enrichment for genes within the top 5% candidate region. (<b>B</b>) KEGG analysis of the selected genes within the candidate region.</p>
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<p>SNP and Indel analysis of the PNLIPRP1 gene in goats. (<b>A</b>) Schematically displaying Motif 1 and Motif 2 of the PNLIPRP1. (<b>B</b>) Gene evolution tree analysis of PNLIPRP1 in different species. (<b>C</b>) SNP and Indels were detected in PNLIPRP1, the loci of which are indicated in the schematic structure of the PNLIPRP1 gene. (<b>D</b>) Electrophoretic profile of amplified PNLIPRP1 26 bp Indel in 2% agarose gel. II genotype: 231 bp; ID genotype: 231 bp and 204 bp; DD genotype: 204 bp. (<b>E</b>) DNA sequencing maps of the 26 bp Indel in the PNLIPRP1 gene. The sequence with the red border shows the difference in sequence fragment.</p>
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<p>Expression pattern of PNLIPRP1 and its role in goat hepatic fat deposition. (<b>A</b>) Protein interaction network diagram constructed using Cytoscape software (version 3.7.2). (<b>B</b>) Expression of PNLIPRP1 in various tissues of 6-month-old goats. (<b>C</b>) Expression levels of fatty acid metabolism-related genes in different genotype populations. (<b>D</b>) PNLIPRP1 26 bp Indel wild-type and mutant-type protein tertiary structure prediction. (<b>E</b>) The effect of PNLIPRP1 26 bp Indel (II genotype or ID genotype) on the expression level of PNLIPRP1 in the liver. (<b>F</b>) HE and ORO staining images of liver with different genotypes; scale bars: 100 µm. Data are presented as the mean ± SEM. <sup>a–c</sup> The difference was significant when data had different letters (<span class="html-italic">p</span> &lt; 0.05), whereas there was no significant difference when data had the same letters (<span class="html-italic">p</span> &gt; 0.05).</p>
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18 pages, 7374 KiB  
Article
Lin28b-let-7 Modulates mRNA Expression of GnRH1 Through Multiple Signaling Pathways Related to Glycolysis in GT1-7 Cells
by Yujing Xie, Xin Li, Meng Wang, Mingxing Chu and Guiling Cao
Animals 2025, 15(2), 120; https://doi.org/10.3390/ani15020120 - 7 Jan 2025
Viewed by 348
Abstract
Lin28b and let-7 miRNA regulate mammalian pubertal initiation and Gonadotropin-releasing hormone (GnRH) production. However, it remains unclear which signaling pathways Lin28b regulates to modulate GnRH production. In this study, the mRNA expression levels of Lin28b and let-7 in the pubertal and juvenile goat [...] Read more.
Lin28b and let-7 miRNA regulate mammalian pubertal initiation and Gonadotropin-releasing hormone (GnRH) production. However, it remains unclear which signaling pathways Lin28b regulates to modulate GnRH production. In this study, the mRNA expression levels of Lin28b and let-7 in the pubertal and juvenile goat hypothalamus and pituitary gland were detected, and Lin28b expression in the pubertal hypothalamus decreased significantly compared with that in juvenile tissues. It was predicted that Lin28b might inhibit GnRH1 expression, which was verified in the GnRH-producing cell model GT1-7 cells. Lin28b inhibited GnRH1 expression and promoted Kiss1/Gpr54 signaling. The pyruvate content and the expression of Hif1a and Hk2, which were related to glycolysis, were also promoted by Lin28b overexpression. Additionally, 77 differentially expressed miRNAs (DEMIs) in Lin28b-overexpressed GT1-7 cells were identified. Bioinformatics analysis and mRNA expression of the target genes of DEMIs revealed that the MAPK and PI3K-AKT-mTOR signaling pathways were key pathways that involved the regulatory effect of Lin28b on GnRH. In GT1-7 cells, GnRH1 expression was suppressed by blocking mTOR signaling with rapamycin, which was rescued by Lin28b overexpression. These results indicate that Lin28b-let-7 regulates GnRH1 expression through several pathways, including the Kiss1/Gpr54, MAPK, and mTOR signaling pathways, which are all related to glucose metabolism and provide new insights into the molecular mechanism of the regulatory role of Lin28b on GnRH production. Full article
(This article belongs to the Section Small Ruminants)
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<p>The diagram of sample collection. Red triangles represent the day that goats were sacrificed. Juv, juvenile group, 1-month-old goats. PE, goats that were in proestrus. E, goats that were in estrus. ME, metestrus goats. DE, goats that were in diestrus.</p>
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<p>The expression of <span class="html-italic">Lin28b</span>, <span class="html-italic">GnRH1 let-7b</span>, and <span class="html-italic">let-7g</span> miRNA in the hypothalamus and (or) pituitary of goats. (<b>a</b>), <span class="html-italic">Lin28b</span> and (<b>b</b>), <span class="html-italic">GnRH1</span> in the hypothalamus. (<b>c</b>,<b>d</b>), <span class="html-italic">let-7b</span> in the hypothalamus and pituitary. (<b>e</b>,<b>f</b>), <span class="html-italic">let-7g</span> in the hypothalamus and pituitary. The different capital letters above the columns chart mean a significant difference with <span class="html-italic">p</span> &lt; 0.01, and the different small letter means a significant difference with <span class="html-italic">p</span> &lt; 0.05 in multiple comparisons. PE, proestrus. E, estrus. ME, metestrus. DE, diestrus. HY, hypothalamus. PI, pituitary.</p>
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<p>The <span class="html-italic">GnRH1</span>, <span class="html-italic">Kiss1</span>, and <span class="html-italic">Gpr54</span> expression and GnRH concentration in Lin28b+ and Lin28b− GT1-7 cells. (<b>a</b>), The <span class="html-italic">Lin28b</span> mRNA relative expression. (<b>b</b>), The GnRH concentration in the supernatant. (<b>c</b>), The <span class="html-italic">GnRH1</span> mRNA relative expression. (<b>d</b>), The relative expression of <span class="html-italic">Kiss1</span> and <span class="html-italic">Gpr54</span>. The different capital letters above the columns chart mean a significant difference with <span class="html-italic">p</span> &lt; 0.01, and the different small letter means a significant difference with <span class="html-italic">p</span> &lt; 0.05 in multiple comparisons.</p>
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<p>The pyruvate and ATP content and mRNA expression of related genes in Lin28b+ and Lin28b− GT1-7 cells. (<b>a</b>), The pyruvate content. (<b>b</b>), The ATP content. (<b>c</b>), The relative expression of <span class="html-italic">Pdk2</span>, <span class="html-italic">Hk2</span> and <span class="html-italic">Hif1a</span>. The different capital letters above the columns chart mean a significant difference with <span class="html-italic">p</span> &lt; 0.01, and the different small letter means a significant difference with <span class="html-italic">p</span> &lt; 0.05 in multiple comparisons.</p>
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<p><span class="html-italic">Lin28b</span> overexpression in GT1-7 cells affected the miRNA expression profile. (<b>a</b>), The classification composition of small RNA in GT1-7 cells. (<b>b</b>), Volcanic map of the DEMIs in GT1-7 cells. (<b>c</b>), Verification of DEMIs. The different capital letters above the columns chart mean significant difference with <span class="html-italic">p</span> &lt; 0.01. (<b>d</b>,<b>e</b>), GO and KEGG enrichment analysis of the target genes of DEMIs.</p>
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<p>The mRNA expression of several target genes of DEMIs and several key genes in the signaling pathway. (<b>a</b>), <span class="html-italic">Map4k3</span>, <span class="html-italic">Map2k4</span>, <span class="html-italic">Mapk9</span>, and <span class="html-italic">JunD</span>. (<b>b</b>), <span class="html-italic">Fgf1</span>, <span class="html-italic">Erbb2</span>, <span class="html-italic">Mapk1</span>, and <span class="html-italic">Atf4</span>. (<b>c</b>), <span class="html-italic">Kcnj3</span>, <span class="html-italic">InsR</span>, <span class="html-italic">Lepr</span>, and <span class="html-italic">Ccnd2</span>. (<b>d</b>), <span class="html-italic">Pik3r1</span>, <span class="html-italic">Akt1</span>, and <span class="html-italic">mTOR</span>. The different capital letters above the columns chart mean a significant difference with <span class="html-italic">p</span> &lt; 0.01, and the different small letter means a significant difference with <span class="html-italic">p</span> &lt; 0.05 in multiple comparisons.</p>
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<p>The mRNA relative expression of <span class="html-italic">GnRH1</span> and genes related to the mTOR signaling pathway in GT1-7 cells with rapamycin treatment and/or <span class="html-italic">Lin28b</span> overexpression. (<b>a</b>), <span class="html-italic">GnRH1</span>. (<b>b</b>,<b>c</b>), <span class="html-italic">InsR</span> and <span class="html-italic">Lepr</span>. (<b>d</b>,<b>e</b>), <span class="html-italic">Pik3r1</span> and <span class="html-italic">Akt1</span>. (<b>f</b>,<b>g</b>), <span class="html-italic">mTOR</span> and <span class="html-italic">S6k</span>. The different capital letters above the columns chart mean a significant difference with <span class="html-italic">p</span> &lt; 0.01, and the different small letter means a significant difference with <span class="html-italic">p</span> &lt; 0.05 in multiple comparisons. Con, control GT1-7 cells; 28b+, the GT1-7 cells with <span class="html-italic">Lin28b</span> overexpression; 200 μM and 500 μM, the GT1-7 cells treated with 200 μM rapamycin or 500 μM rapamycin, respectively; 28b + 200 μM, the GT 1-7 cells treated with 200 μM rapamycin and <span class="html-italic">Lin28b</span> overexpression at the same time; 28b + 500 μM, the GT 1-7 cells treated with 500 μM rapamycin and <span class="html-italic">Lin28b</span> overexpression at the same time.</p>
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<p>The predicted regulatory diagram of <span class="html-italic">Lin28b</span> and glycolysis.</p>
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<p>The overview of <span class="html-italic">Lin28b</span> and the signaling pathways that are involved in GnRH expression.</p>
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16 pages, 1445 KiB  
Article
Identification of Bacillus anthracis Strains from Animal Cases in Ethiopia and Genetic Characterization by Whole-Genome Sequencing
by Abebe Olani, Domenico Galante, Matios Lakew, Bayeta Senbeta Wakjira, Getnet Abie Mekonnen, Tesfaye Rufael, Tsegaye Teklemariam, Wondwosen Kumilachew, Shimalis Dejene, Ayele Woldemeskel, Adanech Wakjira, Getachew Abichu, Baye Ashenafi, Nigatu Kebede, Aklilu Feleke Haile, Fufa Dawo Bari, Laura Del Sambro and Tadesse Eguale
Pathogens 2025, 14(1), 39; https://doi.org/10.3390/pathogens14010039 - 7 Jan 2025
Viewed by 430
Abstract
Anthrax is a zoonotic disease characterized by rapid onset with usual fatal outcomes in livestock and wildlife. In Ethiopia, anthrax is a persistent disease; however, there are limited data on the isolation and molecular characterization of Bacillus anthracis strains. This study aimed to [...] Read more.
Anthrax is a zoonotic disease characterized by rapid onset with usual fatal outcomes in livestock and wildlife. In Ethiopia, anthrax is a persistent disease; however, there are limited data on the isolation and molecular characterization of Bacillus anthracis strains. This study aimed to characterize B. anthracis isolated from animal anthrax outbreaks between 2019 and 2024, from different localities in Ethiopia. B. anthracis was identified using standard microbiology techniques and confirmed by real-time PCR. For the first time in Ethiopia, the genetic diversity of five Bacillus anthracis strains, isolated from dead cattle and goats, was investigated by Whole Genome Sequencing (WGS) and bioinformatics analyses. The five sequenced strains were compared to one Ethiopian B. anthracis genome and the other 29 B. anthracis genomes available in the global genetic databases to determine their phylogeny. The genomes of the strains were also analyzed to detect the presence of antimicrobial resistance and virulence genes. The whole genome SNP analysis showed that the Ethiopian B. anthracis strains were grouped in the A clade. Three strains (BA2, BA5, and BA6) belonged to the A.Br.034 subgroup (A.Br.005/006), and two strains (BA1 and BA4) belonged to the A.Br.161 (Heroin) clade of the Trans-Eurasian (TEA) group. The findings of this study will contribute to expanding the current understanding of the anthrax hotspots in Ethiopia, and the phylogenetic correlation and/or diversity of the circulating strains. Full article
(This article belongs to the Special Issue Current Research on Bacillus anthracis Infection)
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<p>Map of Ethiopia showing the sites where <span class="html-italic">B. anthracis</span> strains were isolated.</p>
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<p>Ox dead of anthrax showing bloody discharge from the anus in Bonga Town (<b>A</b>); burying of a cow dead of anthrax in Deber Work Town, 2023 (<b>B</b>); bones collected following the death of a cow in Ada Berga District (<b>C</b>).</p>
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<p>Evidence of Gram-positive, thick long chains of bacilli (<b>A</b>); evidence of <span class="html-italic">B. anthracis</span> spores (in green) by malachite green staining (<b>B</b>); evidence of capsule pinkish-red stained, and bacilli blue stained by Giemsa staining (<b>C</b>).</p>
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<p>(<b>A</b>) Global phylogeny of 35 <span class="html-italic">B. anthracis</span> strains generated using the SNP Phylogeny (Samtools) Pipeline, with strains grouped by CanSNP classifications as defined [<a href="#B18-pathogens-14-00039" class="html-bibr">18</a>]; (<b>B</b>) Phylogenetic tree of representatives from the A.Br.034 (Ancient A) CanSNP group, and (<b>C</b>) Phylogenetic tree of representatives from the A.Br.161 (Heroin) CanSNP group. The reference genome used was ‘Ames Ancestor’ (NC_007530.2). Colored tips indicate CanSNP group nicknames, while empty circles represent strains that have not yet been assigned to a genetic group.</p>
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12 pages, 1751 KiB  
Article
Chronic Dexamethasone Disturbs the Circadian Rhythm of Melatonin and Clock Genes in Goats
by Liuping Cai, Qu Chen, Canfeng Hua, Liqiong Niu, Qijun Kong, Lei Wu and Yingdong Ni
Animals 2025, 15(1), 115; https://doi.org/10.3390/ani15010115 - 6 Jan 2025
Viewed by 479
Abstract
Dex is a drug commonly used as an immunosuppressive and anti-inflammatory agent in humans and animals. GCs have a profound impact on melatonin expression and biological rhythm. However, the effect of chronic exposure to Dex on melatonin secretion and biological clock gene expression [...] Read more.
Dex is a drug commonly used as an immunosuppressive and anti-inflammatory agent in humans and animals. GCs have a profound impact on melatonin expression and biological rhythm. However, the effect of chronic exposure to Dex on melatonin secretion and biological clock gene expression in ruminants is still unclear. Ten goats were randomly divided into two groups: the control group was injected with saline, and the Dex-treated group was intramuscularly injected daily for 21 d with 0.2 mg/kg Dex. The rhythm of melatonin secretion in the plasma was disturbed in the Dex group, and the plasma and colon levels of melatonin were lower in the Dex group compared to the control group (p < 0.05). Dex leads to a significant decrease in the expression of Arylalkylamine N-acetyltransferase (AANAT), a key melatonin synthase, in the pineal gland and colon. Detecting intestinal leakage-related indices showed that diamine oxidase (DAO) and lipopolysaccharide (LPS) content increased significantly in the Dex group (p < 0.05). We also detected genes associated with biological rhythms in the plasma. In the control group, the five tested genes showed circadian rhythms, but the circadian rhythms of Clock, Cry1, Cry2, and Per2 were abolished or blunted by the Dex (p < 0.05). Protein levels of CLOCK and BMAL1 in the colon changed significantly (p < 0.05). In conclusion, the above experimental results show that chronic exposure to Dex leads to the disorder of the circadian rhythms of melatonin secretion and clock genes. Full article
(This article belongs to the Section Animal Physiology)
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<p>Effect of chronic Dex exposure on the melatonin and cortisol content. (<b>A</b>) The concentration of melatonin in the plasma; (<b>B</b>) the concentration of cortisol in the plasma. (<b>C</b>) The melatonin content in the colon; (<b>D</b>) the level of AANTA in the pineal gland; (<b>E</b>) the level of AANTA in the colon. n = 5 goats per time point. The degree of fitting is represented by the R<sup>2</sup> values. Data are mean ± SEM; * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 compared with Con.</p>
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<p>The circadian rhythm parameters of clock genes in the plasma. (<b>A</b>) Clock genes; (<b>B</b>) Cry1 gene; (<b>C</b>) Cry2 gene; (<b>D</b>) Per2 gene; (<b>E</b>) Per3 gene. In the graphs, the data markers denote the mRNA expression levels of clock genes. The curves illustrate the 24 h period identified by cosinor analysis. There are 5 goats for each time point (n = 5). The R<sup>2</sup> values signify the degree of fitting. The values are in the form of mean ± SEM.</p>
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<p>Effect of chronic Dex exposure on the concentration of GSH and MDA in the plasma. (<b>A</b>) GSH content; (<b>B</b>) MDA content. n = 5 goats, and the values are mean ± SEM; * <span class="html-italic">p</span> &lt; 0.05 compared with Con.</p>
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<p>Effect of chronic Dex exposure on the concentration of DAO and LPS in the plasma. (<b>A</b>) DAO content; (<b>B</b>) LPS content. n = 5 goats, and the values are mean ± SEM, * <span class="html-italic">p</span> &lt; 0.05 compared with Con.</p>
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<p>Effect of chronic Dex exposure on the protein expression of CLOCK, BMAL1, and GR. (<b>A</b>) CLOCK protein expression in the colon; (<b>B</b>) BMAL1 protein expression in the colon; (<b>C</b>) GR protein expression in the colon; (<b>D</b>) GR protein expression in the cecum. n = 5 goats, and the values are mean ± SEM; compared with the Con, * <span class="html-italic">p</span> &lt; 0.05.</p>
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12 pages, 251 KiB  
Article
Seasonal and Altitudinal Effects on Chemical Composition and Rumen Degradability of Blackberry Leaves in Northwestern Italian Alps
by Sonia Tassone, Salvatore Barbera, Sara Glorio Patrucco, Hatsumi Kaihara and Khalil Abid
Animals 2025, 15(1), 111; https://doi.org/10.3390/ani15010111 - 6 Jan 2025
Viewed by 424
Abstract
The blackberry poses a threat as an invasive plant in various regions worldwide, where it aggressively competes with native species and risks delicate ecosystems. Livestock grazing has emerged as a potential strategy to mitigate its spread. This study investigated the effects of seasonal [...] Read more.
The blackberry poses a threat as an invasive plant in various regions worldwide, where it aggressively competes with native species and risks delicate ecosystems. Livestock grazing has emerged as a potential strategy to mitigate its spread. This study investigated the effects of seasonal variations and altitude on the chemical composition and in vitro degradability of blackberry leaves. The leaves accessible to goats were collected in the Northwestern Italian Alps across all seasons at three altitudes (low: 450 m, medium: 700 m, high: 1000 m). The findings indicated that blackberry leaves can serve as a cost-effective, high-protein, and high-fiber feed for ruminants in these regions. However, the goats exhibited a limited capacity for fiber degradation. Furthermore, the nutritional value of these leaves was significantly influenced by the season, altitude, and their interaction. Spring leaves had the highest protein content (241.9 g/kg dry matter) and the best digestibility, with lower lignin levels (69 g/kg dry matter). At higher altitudes, lignin content decreased significantly during winter compared to the other seasons, resulting in a significant increase in fiber degradability. These insights offer crucial guidance for optimizing the utilization of the blackberry plant in goat feeding systems and underscores the necessity of considering both seasonal and altitudinal factors in grazing management practices. Full article
30 pages, 4295 KiB  
Article
Characterisation of Staphylococcus aureus Strains and Their Prophages That Carry Horse-Specific Leukocidin Genes lukP/Q
by Stefan Monecke, Sindy Burgold-Voigt, Andrea T. Feßler, Martina Krapf, Igor Loncaric, Elisabeth M. Liebler-Tenorio, Sascha D. Braun, Celia Diezel, Elke Müller, Martin Reinicke, Annett Reissig, Adriana Cabal Rosel, Werner Ruppitsch, Helmut Hotzel, Dennis Hanke, Christiane Cuny, Wolfgang Witte, Stefan Schwarz and Ralf Ehricht
Toxins 2025, 17(1), 20; https://doi.org/10.3390/toxins17010020 - 3 Jan 2025
Viewed by 602
Abstract
Leukocidins of Staphylococcus (S.) aureus are bicomponent toxins that form polymeric pores in host leukocyte membranes, leading to cell death and/or triggering apoptosis. Some of these toxin genes are located on prophages and are associated with specific hosts. The genes lukP/Q [...] Read more.
Leukocidins of Staphylococcus (S.) aureus are bicomponent toxins that form polymeric pores in host leukocyte membranes, leading to cell death and/or triggering apoptosis. Some of these toxin genes are located on prophages and are associated with specific hosts. The genes lukP/Q have been described from equine S. aureus isolates. We examined the genomes, including the lukP/Q prophages, of S. aureus strains belonging to clonal complexes CC1, CC350, CC816, and CC8115. In addition to sequencing, phages were characterised by mitomycin C induction and transmission electron microscopy (TEM). All lukP/Q prophages integrated into the lip2=geh gene, and all included also the gene scn-eq encoding an equine staphylococcal complement inhibitor. The lukP/Q prophages clustered, based on gene content and allelic variants, into three groups. One was found in CC1 and CC97 sequences; one was present mainly in CC350 but also in other lineages (CC1, CC97, CC133, CC398); and a third one was exclusively observed in CC816 and CC8115. Prophages of the latter group additionally included a rare enterotoxin A allele (sea320E). Moreover, a prophage from a CC522 goat isolate was found to harbour lukP. Its lukF component could be regarded as chimaera comprising parts of lukQ and of lukF-P83. A putative kinase gene of 1095 basepairs was found to be associated with equine strains of S. aureus. It was also localised on prophages. However, these prophages were different from the ones that carried lukP/Q, and three different integration sites of kinase-carrying phages were identified. These observations confirmed the presence of prophage-located important virulence-associated genes in equine S. aureus and that certain prophages might determine the host specificity of the staphylococcal strains they reside in. Full article
(This article belongs to the Section Bacterial Toxins)
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Figure 1

Figure 1
<p>(<b>a</b>) Alignment of <span class="html-italic">lukQ</span> and <span class="html-italic">lukF-P83</span>, showing that sequences from CP138360, pubMLST ID 24359, and JXHY01000064 match the sequence of <span class="html-italic">lukQ</span> until position 702 and the one of <span class="html-italic">lukF</span>-PV83 henceforth. (<b>b</b>) Alignment of <span class="html-italic">lukP/lukM</span> sequences, showing that the sequences from CP138360, pubMLST ID 24359, and JXHY01000064 match the sequence of equine <span class="html-italic">lukP</span> rather than the one of bovine <span class="html-italic">lukM</span>.</p>
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<p>(<b>a</b>) Alignment of <span class="html-italic">lukQ</span> and <span class="html-italic">lukF-P83</span>, showing that sequences from CP138360, pubMLST ID 24359, and JXHY01000064 match the sequence of <span class="html-italic">lukQ</span> until position 702 and the one of <span class="html-italic">lukF</span>-PV83 henceforth. (<b>b</b>) Alignment of <span class="html-italic">lukP/lukM</span> sequences, showing that the sequences from CP138360, pubMLST ID 24359, and JXHY01000064 match the sequence of equine <span class="html-italic">lukP</span> rather than the one of bovine <span class="html-italic">lukM</span>.</p>
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<p>Transmission electron micrographs of phage particles from the preparation of strain IMT39173. (<b>A</b>), icosahedral phage. (<b>B</b>), prolate phage with large head diameter and oval shape. (<b>C</b>), prolate phage with small head diameter and angular shape. Negative contrast preparation with uranyl acetate. Size bars = 100 nm.</p>
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<p>Transmission electron micrographs of phage particles from the preparation of strain IMT37083. (<b>A</b>), icosahedral phage. (<b>B</b>), phage with mildly elongated head. Negative contrast preparation with uranyl acetate. Size bars = 100 nm.</p>
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<p>Transmission electron micrographs of phage particles from the preparation of strain V353. (<b>A</b>), prolate phage with large head diameter and oval shape. (<b>B</b>), prolate phage with small head diameter and angular shape. Negative contrast preparation with uranyl acetate. Size bars = 100 nm.</p>
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<p>Transmission electron micrograph of incomplete phage particles from the preparation of strain V641. (<b>A</b>), two prolate phage particles, one with small head diameter and angular shape (1) and one with large head diameter and oval shape (2). (<b>B</b>), head of an icosahedral phage particle. Negative contrast preparation with uranyl acetate. Size bars = 100 nm.</p>
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<p>Transmission electron micrographs of phage particles from the preparation of strain 17CS1042. (<b>A</b>), large icosahedral phage. (<b>B</b>), small icosahedral phage. Negative contrast preparation with uranyl acetate. Size bars = 100 nm.</p>
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