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24 pages, 3149 KiB  
Article
Evaluation of Aggregate Oral Fluid Sampling for Early Detection of African Swine Fever Virus Infection
by Bonto Faburay, Kathleen O’Hara, Marta Remmenga, Theophilus Odoom, Sherry Johnson, William Tasiame, Matilda Ayim-Akonor, Benita Anderson, Kingsley Kwabena Amoako, Diane Holder, Wu Ping, Michelle Zajac, Vivian O’Donnell, Lizhe Xu, Robin Holland, Corrie Brown, Randall Levings and Suelee Robbe-Austerman
Viruses 2025, 17(8), 1089; https://doi.org/10.3390/v17081089 - 6 Aug 2025
Abstract
African swine fever (ASF) needs to be controlled, and prevention of the spread of African swine fever virus (ASFV) is dependent on enhanced surveillance and early disease detection. Commercial swine operations, especially in North America, Europe, and Asia, are characterized by comparatively large [...] Read more.
African swine fever (ASF) needs to be controlled, and prevention of the spread of African swine fever virus (ASFV) is dependent on enhanced surveillance and early disease detection. Commercial swine operations, especially in North America, Europe, and Asia, are characterized by comparatively large numbers of pigs, and sampling individual pigs, which represents the main strategy for current ASF surveillance, can be both costly and labor intensive. A study performed in Ghana was designed to estimate the diagnostic sensitivity of pen-based aggregate oral fluid testing for ASFV in infected pigs in a pen of 30 animals and to evaluate its utility as a tool to support surveillance of ASF in the US. This study was performed in three phases: (i) virus (Ghana ASFV24) amplification in a target host species to generate the challenge inoculum; (ii) titration of the inoculum (10% spleen homogenate) in target host species to determine the minimum dose inducing acute ASF in pigs with survival up to 5–6 days post-inoculation (dpi); and (iii) the main study, involving 186 pigs, consisting of 6 replicates of 30 pigs per pen and one seeder pig inoculated with wildtype ASFV (highly virulent genotype II) per pen. Daily sampling of aggregate oral fluids, uncoagulated blood, oropharyngeal swabs, fecal and water nipple swabs, and recording of rectal temperatures and clinical observations was carried out. The seeder pigs were each inoculated intramuscularly with 0.5 mL of the 10% spleen homogenate, which induced the desired clinical course of ASF in the pigs, with survival of up to 6 dpi. ASFV DNA was detected in the seeder pigs as early as 1 dpi and 2 dpi in the blood and oropharyngeal swabs, respectively. Transmission of ASFV from the seeder pigs to the contact pig population was detected via positive amplification of ASFV DNA in aggregate oral fluid samples at 3 days post-contact (dpc) in 4 out of 6 pens, and in all 6 pens, at 4 dpc. Testing of oropharyngeal swabs and blood samples from individual pigs revealed a variable number of ASFV-positive pigs between 3 and 5 dpc, with detection of 100% positivity between 6 and 18 dpc, the study endpoint. These findings demonstrate the potential utility of aggregate oral fluid sampling for sensitive and early detection of ASFV incursion into naïve swine herds. It also demonstrates that testing of environmental samples from the premises could further enhance overall ASF early detection and surveillance strategies. Full article
(This article belongs to the Collection African Swine Fever Virus (ASFV))
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20 pages, 450 KiB  
Article
Four Organic Protein Source Alternatives to Fish Meal for Pacific White Shrimp (Penaeus vannamei) Feeding
by Yosu Candela-Maldonado, Imane Megder, Eslam Tefal, David S. Peñaranda, Silvia Martínez-Llorens, Ana Tomás-Vidal, Miguel Jover-Cerdá and Ignacio Jauralde
Fishes 2025, 10(8), 384; https://doi.org/10.3390/fishes10080384 - 5 Aug 2025
Viewed by 39
Abstract
The use of eco-organic ingredients as a source of protein in aquaculture diets needs important attention due to the growing demand for organic seafood products. The present study evaluated the effects of fish meal substitution by different organic ingredients on the growth, body [...] Read more.
The use of eco-organic ingredients as a source of protein in aquaculture diets needs important attention due to the growing demand for organic seafood products. The present study evaluated the effects of fish meal substitution by different organic ingredients on the growth, body composition, retention efficiency, enzyme activity, and nutrient digestibility of white shrimp Penaeus vannamei. The four dietary formulations tested were formulated with organic ingredients and the fish meal was replaced by the following organic protein meals: Iberian pig viscera meal (PIG), trout by-product meal (TRO), insect meal (FLY), and organic vegetable meal (WHT), in addition to a control diet (CON) that included 15% fish meal. A growth trial was carried out for 83 days, raising 1 g shrimp to commercial size (20 g). Shrimp were stocked at 167 shrimp/m3 (15 individuals per 90 L tank). The results showed that the growth obtained by shrimp fed with TRO (19.27 g) and PIG (19.35 g) were similar in weight gain to the control diet (20.76 g), while FLY (16.04 g) and WHT (16.73 g) meals resulted in a significant lower final weight. The FLY diet showed significantly lower protein digestibility (68.89%) compared to the CON, PIG, TRO, and WHT diets, and significantly higher trypsin activity (0.17 mU/g) compared to shrimp fed with the PIG, TRO, and WHT diets. Shrimp fed with WHT have a significantly lower body weight percentage of protein (19.69%) than shrimp fed with the WHT and TRO diets, and some significant differences in dietary aminoacidic levels affecting amino acid body composition. These results indicate that Iberian pig viscera and trout by-product meal can successfully replace fish meal in Pacific white shrimp aquaculture. Full article
(This article belongs to the Special Issue Advances in Aquaculture Feed Additives)
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23 pages, 5970 KiB  
Review
Practical Review on Aetio-Pathogenesis and Symptoms in Pigs Affected by Clinical and Subclinical Oedema Disease and the Use of Commercial Vaccines Under Field Conditions
by Juan Hernandez-Garcia, Isaac Ballarà Rodriguez, Ramon Jordà Casadevall, Sergi Bruguera, David Llopart and Emili Barba-Vidal
Animals 2025, 15(15), 2275; https://doi.org/10.3390/ani15152275 - 4 Aug 2025
Viewed by 195
Abstract
The impact of Oedema Disease produced by Shiga toxigenic Escherichia coli (STEC) in swine is increasing in some production countries due to increasing limitations on treatment with antimicrobials and zinc oxide, either because of the increased prevalence of multi-resistant strains or because of [...] Read more.
The impact of Oedema Disease produced by Shiga toxigenic Escherichia coli (STEC) in swine is increasing in some production countries due to increasing limitations on treatment with antimicrobials and zinc oxide, either because of the increased prevalence of multi-resistant strains or because of legal restrictions. The main pathological effect of Shiga toxin 2e is represented by damage to the endothelial cells of the blood vessel walls, leading to liquid extravasation and oedema formation in multiple tissues. These oedemas are generally easily identifiable in acute clinical cases. However, disease caused by Shiga toxin can occur without any externally visible oedema in the pigs, as observed in the subclinical presentation of Oedema Disease. It also causes productive losses, so it is important to identify and/or diagnose cases to set up control measures in order to optimize production and health. This article includes a comprehensive review of lesions and signs caused by Shiga toxin toxicosis in pigs, as well as other insights about the aetiology and epidemiology of STEC in pigs, and the effect of Shiga toxin recombinant toxoid vaccines in reducing these clinical and subclinical signs under field conditions. Full article
(This article belongs to the Section Pigs)
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10 pages, 588 KiB  
Article
Genome-Wide Association Study of Gluteus Medius Muscle Size in a Crossbred Pig Population
by Yu He, Chunyan Bai, Junwen Fei, Juan Ke, Changyi Chen, Xiaoran Zhang, Wuyang Liu, Jing Li, Shuang Liang, Boxing Sun and Hao Sun
Vet. Sci. 2025, 12(8), 730; https://doi.org/10.3390/vetsci12080730 - 3 Aug 2025
Viewed by 107
Abstract
The size of the gluteus medius muscle (GM) in swine significantly impacts both hindlimb conformation and carcass yield, while little is known about the genetic architecture of this trait. This study aims to estimate genetic parameters and identify candidate genes associated with this [...] Read more.
The size of the gluteus medius muscle (GM) in swine significantly impacts both hindlimb conformation and carcass yield, while little is known about the genetic architecture of this trait. This study aims to estimate genetic parameters and identify candidate genes associated with this trait through a genome-wide association study (GWAS). A total of 439 commercial crossbred pigs, possessing both Landrace and Yorkshire ancestry, were genotyped using the Porcine 50K chip. The length and width of the GM were directly measured, and the area was then calculated from these values. The heritabilities were estimated by HIBLUP (V1.5.0) software, and the GWAS was conducted employing the BLINK model implemented in GAPIT3. The heritability estimates for the length, width, and area of the GM were 0.43, 0.40, and 0.46, respectively. The GWAS identified four genome-wide significant SNPs (rs81381267, rs697734475, rs81298447, and rs81458910) associated with the gluteus medius muscle area. The PDE4D gene was identified as a promising candidate gene potentially involved in the regulation of gluteus medius muscle development. Our analysis revealed moderate heritability estimates for gluteus medius muscle size traits. These findings enhance our understanding of the genetic architecture underlying porcine muscle development. Full article
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25 pages, 1990 KiB  
Article
Fecal and Environmental Shedding of Influenza A Virus in Brazilian Swine: Genomic Evidence of Recent Human-to-Swine Transmission
by Nágila Rocha Aguilar, Beatriz Senra Alvares da Silva Santos, Bruno Zinato Carraro, Brenda Monique Magalhães Rocha, Jardelina de Souza Todao Bernardino, Ana Luiza Soares Fraiha, Alex Ranieri Jeronimo Lima, Gabriela Ribeiro, Alessandra Silva Dias, Renata Rezende Carvalho, Bruna Ferreira Sampaio Ribeiro, Marta Giovanetti, Luiz Carlos Júnior Alcântara, Sandra Coccuzzo Sampaio, Maria Carolina Quartim Barbosa Elias Sabbaga, Rafael Romero Nicolino, Zélia Inês Portela Lobato, Maria Isabel Maldonado Coelho Guedes, Cesar Rossas Mota Filho, Vincent Louis Viala, Bruna Coelho Lopes and Erica Azevedo Costaadd Show full author list remove Hide full author list
Pathogens 2025, 14(8), 753; https://doi.org/10.3390/pathogens14080753 - 31 Jul 2025
Viewed by 261
Abstract
Surveillance of swine influenza A virus (swIAV) traditionally focuses on respiratory matrices, yet emerging evidence suggests that fecal shedding and secondary environmental contamination may also contribute to viral dissemination. In this study, we collected and analyzed nasal, rectal, environmental, milk, and colostrum samples [...] Read more.
Surveillance of swine influenza A virus (swIAV) traditionally focuses on respiratory matrices, yet emerging evidence suggests that fecal shedding and secondary environmental contamination may also contribute to viral dissemination. In this study, we collected and analyzed nasal, rectal, environmental, milk, and colostrum samples from naturally infected pigs in a commercial farm in Minas Gerais, Brazil. IAV RNA was detected in 25% of samples, including 42% from asymptomatic animals, with nasal swabs showing higher detection rates (30%) than rectal swabs (20%), though rectal Ct values were consistently higher, indicative of lower viral loads. We successfully isolated viable viruses from feces and effluent samples. Whole-genome sequencing revealed co-circulation of enzootic pH1N1 clade #2 (HA) and pN1 clade #4 (NA), alongside human-origin H3N2 sequences clustering within clade 3C.2a1b.2a.2a.1, and N2 segments related to pre-3C human lineages from 2001 to 2002. Phylogenetic and p-distance analyses support both recent reverse zoonosis and historical transmission events. Detection of complete HA/NA sequences from rectal swabs and treated effluent further emphasizes the surveillance value of non-respiratory matrices. The integration of respiratory and fecal/environmental sampling appears important to achieve more comprehensive IAV monitoring in swine herds and may have significant implications for One Health strategies in Brazil and beyond. Full article
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16 pages, 2491 KiB  
Article
High-Yield Production of PCV2 Cap Protein: Baculovirus Vector Construction and Cultivation Process Optimization
by Long Cheng, Denglong Xie, Wei Ji, Xiaohong Ye, Fangheng Yu, Xiaohui Yang, Nan Gao, Yan Zhang, Shu Zhu and Yongqi Zhou
Vaccines 2025, 13(8), 801; https://doi.org/10.3390/vaccines13080801 - 28 Jul 2025
Viewed by 335
Abstract
Background/Objectives: Porcine circovirus type 2 (PCV2) infection causes porcine circovirus disease (PCVD), a global immunosuppressive disease in pigs. Its clinical manifestations include post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS), which cause significant economic losses to the swine industry. [...] Read more.
Background/Objectives: Porcine circovirus type 2 (PCV2) infection causes porcine circovirus disease (PCVD), a global immunosuppressive disease in pigs. Its clinical manifestations include post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS), which cause significant economic losses to the swine industry. The Cap protein, which is the major protective antigen of PCV2, can self-assemble to form virus-like particles (VLPs) in the insect baculovirus expression system. Few studies have compared the expression of Cap proteins in different baculovirus expression systems. Methods: In this study, we compared two commonly commercialized baculovirus construction systems with the Cap protein expression in various insect cells. Results: The results demonstrate that the flashBAC system expressed the Cap protein at higher levels than the Bac-to-Bac system. Notably, when expressing four copies of the Cap protein, the flashBAC system achieved the highest protein yield in High Five cells, where it reached 432 μg/mL at 5 days post-infection (dpi) with 27 °C cultivation. Animal experiments confirmed that the purified Cap protein effectively induced specific antibody production in mice and swine. Conclusions: This study provides critical data for optimizing the production of the PCV2 Cap protein, which is of great significance for reducing the production cost of PCV2 vaccines and improving the industrial production efficiency. Full article
(This article belongs to the Section Veterinary Vaccines)
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25 pages, 3631 KiB  
Article
Prebiotic Xylo-Oligosaccharides Modulate the Gut Microbiome to Improve Innate Immunity and Gut Barrier Function and Enhance Performance in Piglets Experiencing Post-Weaning Diarrhoea
by James S. Stanley, Stephen C. Mansbridge, Michael R. Bedford, Ian F. Connerton and Kenneth H. Mellits
Microorganisms 2025, 13(8), 1760; https://doi.org/10.3390/microorganisms13081760 - 28 Jul 2025
Viewed by 444
Abstract
During commercial pig production, weaning is a major stressor that disrupts the gut microbiome, compromises intestinal barrier integrity, and increases the susceptibility of piglets to pathogens. This often results in post-weaning diarrhoea (PWD), leading to growth retardation, morbidity, and economic loss. This study [...] Read more.
During commercial pig production, weaning is a major stressor that disrupts the gut microbiome, compromises intestinal barrier integrity, and increases the susceptibility of piglets to pathogens. This often results in post-weaning diarrhoea (PWD), leading to growth retardation, morbidity, and economic loss. This study investigated the effects of dietary xylo-oligosaccharide (XOS) supplementation on the growth performance and gut health of 216 piglets with naturally occurring PWD. Piglets received either 0 (CON), 50 (XOS-50), or 500 (XOS-500) mg XOS/kg feed from weaning at 28 days of age (d1) for 54 days. XOS-500 significantly improved body weight at d22 and d54, but had no effect on average daily gain, daily feed intake (DFI), or feed conversion ratio. The intestinal microbiota alpha-diversity was unaffected by XOS, though jejunal beta diversity differed between CON and XOS-500 groups at d22. Jejunal Chao richness correlated positively with d54 body weight, while ileal Chao richness correlated negatively with DFI. Salmonella was present in all diet groups but did not differ in abundance; however, the levels were negatively correlated with alpha diversity. XOSs increased Lactobacillus (d22, d54) and Clostridium_XI (d22), while reducing Veillonellaceae spp. (d22). XOSs reduced jejunal goblet cell (GC) density at d22 but increased duodenal and jejunal GCs and reduced duodenal crypt depth at d54. XOSs upregulated the genes for the tight junction proteins CLDN2, CLDN3, ALPI, and ZO-1, while downregulating the cytokine IL-8. These findings highlight XOSs’ potential to improve growth and gut health in weaning piglets with naturally occurring PWD, to maintain productivity and enhance welfare. Full article
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36 pages, 528 KiB  
Review
Advancements in Modern Nucleic Acid-Based Multiplex Testing Methodologies for the Diagnosis of Swine Infectious Diseases
by Jingneng Wang, Lei Zhou and Hanchun Yang
Vet. Sci. 2025, 12(8), 693; https://doi.org/10.3390/vetsci12080693 - 24 Jul 2025
Viewed by 293
Abstract
Swine infectious diseases, often caused by multiple co-infecting agents, pose severe global threats to pig health and industry economics. Conventional single-plex testing assays, whether relying on pathogen antigens or nucleic acids, exhibit limited efficacy in the face of co-infection events. The modern nucleic [...] Read more.
Swine infectious diseases, often caused by multiple co-infecting agents, pose severe global threats to pig health and industry economics. Conventional single-plex testing assays, whether relying on pathogen antigens or nucleic acids, exhibit limited efficacy in the face of co-infection events. The modern nucleic acid-based multiplex testing (NAMT) methods demonstrate substantial strengths in the simultaneous detection of multiple pathogens involving co-infections owing to their remarkable sensitivity, exceptional specificity, high-throughput, and short turnaround time. The development, commercialization, and application of NAMT assays in swine infectious disease surveillance would be advantageous for early detection and control of pathogens at the onset of an epidemic, prior to community transmission. Such approaches not only contribute to saving the lives of pigs but also aid pig farmers in mitigating or preventing substantial economic losses resulting from infectious disease outbreaks, thereby alleviating unwanted pressure on animal and human health systems. The current literature review provides an overview of some modern NAMT methods, such as multiplex quantitative real-time PCR, multiplex digital PCR, microarrays, microfluidics, next-generation sequencing, and their applications in the diagnosis of swine infectious diseases. Furthermore, the strengths and weaknesses of these methods were discussed, as well as their future development and application trends in swine disease diagnosis. Full article
(This article belongs to the Special Issue Exploring Innovative Approaches in Veterinary Health)
15 pages, 2688 KiB  
Article
Recombinant Tetrameric Neuraminidase Subunit Vaccine Provides Protection Against Swine Influenza A Virus Infection in Pigs
by Ao Zhang, Bin Tan, Jiahui Wang and Shuqin Zhang
Vaccines 2025, 13(8), 783; https://doi.org/10.3390/vaccines13080783 - 23 Jul 2025
Viewed by 359
Abstract
Background/Objectives: Swine influenza A virus (swIAV), a prevalent respiratory pathogen in porcine populations, poses substantial economic losses to global livestock industries and represents a potential threat to public health security. Neuraminidase (NA) has been proposed as an important component for universal influenza [...] Read more.
Background/Objectives: Swine influenza A virus (swIAV), a prevalent respiratory pathogen in porcine populations, poses substantial economic losses to global livestock industries and represents a potential threat to public health security. Neuraminidase (NA) has been proposed as an important component for universal influenza vaccine development. NA has potential advantages as a vaccine antigen in providing cross-protection, with specific antibodies that have a broad binding capacity for heterologous viruses. In this study, we evaluated the immunogenicity and protective efficacy of a tetrameric recombinant NA subunit vaccine in a swine model. Methods: We constructed and expressed structurally stable soluble tetrameric recombinant NA (rNA) and prepared subunit vaccines by mixing with ISA 201 VG adjuvant. The protective efficacy of rNA-ISA 201 VG was compared to that of a commercial whole inactivated virus vaccine. Pigs received a prime-boost immunization (14-day interval) followed by homologous viral challenge 14 days post-boost. Results: Both rNA-ISA 201 VG and commercial vaccine stimulated robust humoral responses. Notably, the commercial vaccine group exhibited high viral-binding antibody titers but very weak NA-specific antibodies, whereas rNA-ISA 201 VG immunization elicited high NA-specific antibody titers alongside substantial viral-binding antibodies. Post-challenge, both immunization with rNA-ISA 201 VG and the commercial vaccine were effective in inhibiting viral replication, reducing viral load in porcine respiratory tissues, and effectively mitigating virus-induced histopathological damage, as compared to the PBS negative control. Conclusions: These findings found that the anti-NA immune response generated by rNA-ISA 201 VG vaccination provided protection comparable to that of a commercial inactivated vaccine that primarily induces an anti-HA response. Given that the data are derived from one pig per group, there is a requisite to increase the sample size for more in-depth validation. This work establishes a novel strategy for developing next-generation SIV subunit vaccines leveraging NA as a key immunogen. Full article
(This article belongs to the Special Issue Vaccine Development for Swine Viral Pathogens)
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15 pages, 1363 KiB  
Article
The Effects of Dietary Supplementation of Chestnut Tannic Acid on the Growth Performance, Gut Morphology and Microbiota of Weaned Piglets
by Jinzhou Zhang, Yuting Zhang, Yuya Wang, Yanwei Li, Dongyang Liu, Hongbing Xie, Yongqiang Wang, Meinan Chang, Liping Guo and Zhiguo Miao
Metabolites 2025, 15(7), 477; https://doi.org/10.3390/metabo15070477 - 15 Jul 2025
Viewed by 369
Abstract
Background/Objectives: This study investigated the effects of chestnut tannic acid (TA) on the growth performance, the expression of tight junction proteins and the composition of the gut microbiota of weaned piglets, which could provide novel insights into the application of TA in [...] Read more.
Background/Objectives: This study investigated the effects of chestnut tannic acid (TA) on the growth performance, the expression of tight junction proteins and the composition of the gut microbiota of weaned piglets, which could provide novel insights into the application of TA in swine production. Methods: In a 42-day trial, 180 healthy, 21-day-old Duroc × Landrace × Yorkshire piglets were randomly assigned to a Control group and four treatment groups (TA1–4), fed commercial diets supplemented with 0, 0.06%, 0.12%, 0.18% or 0.24% TA. Each group had six replicates of six pigs each. Results: The average daily gain in all TA groups, the jejunal and ileal villus height and the villus height-to-crypt depth ratio in the TA3 and TA4 groups were markedly increased (p < 0.05). The mRNA levels of MUC2 and ZO-1 were upregulated in the TA3 group, as were those of MUC4 in the jejunum and ileum and claudin in the duodenum and ileum; glutathione peroxidase and total antioxidant capacity were upregulated in the duodenum and jejunum in the TA3 group, and total superoxide dismutase was increased in all the TA2 groups (p < 0.05). Conversely, the malondialdehyde significantly decreased in all the TA groups (p < 0.05). TA supplementation improved the alpha diversity of the intestinal microflora and augmented probiotic abundance while reducing that of pathogenic bacteria. The contents of acetic, isobutyric, valeric, isovaleric, hexanoic and propionic acids, as well as total short-chain fatty acids (SCFA), were higher in the TA2 and TA3 groups (p < 0.05). Conclusions: TA inclusion in piglet diets improved the intestinal environment by upregulating the antioxidant enzymes, improving intestinal morphology and promoting probiotic growth and SCFA production while reducing pathogenic bacterial abundance, consequently enhancing the gut barrier and the growth of weaned piglets. Full article
(This article belongs to the Section Animal Metabolism)
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23 pages, 22555 KiB  
Article
Citrate Transporter Expression and Localization: The Slc13a5Flag Mouse Model
by Jan C.-C. Hu, Tian Liang, Hong Zhang, Yuanyuan Hu, Yasuo Yamakoshi, Ryuji Yamamoto, Chuhua Zhang, Hui Li, Charles E. Smith and James P. Simmer
Int. J. Mol. Sci. 2025, 26(14), 6707; https://doi.org/10.3390/ijms26146707 - 12 Jul 2025
Viewed by 361
Abstract
The sodium–citrate cotransporter (NaCT) plays a crucial role in citrate transport during amelogenesis. Mutations in the SLC13A5 gene, which encodes the NaCT, cause early infantile epileptic encephalopathy 25 and amelogenesis imperfecta. We analyzed developing pig molars and determined that the citrate concentrations in [...] Read more.
The sodium–citrate cotransporter (NaCT) plays a crucial role in citrate transport during amelogenesis. Mutations in the SLC13A5 gene, which encodes the NaCT, cause early infantile epileptic encephalopathy 25 and amelogenesis imperfecta. We analyzed developing pig molars and determined that the citrate concentrations in secretory- and maturation-stage enamel are both 5.3 µmol/g, with about 95% of the citrate being bound to mineral. To better understand how citrate might enter developing enamel, we developed Slc13a5Flag reporter mice that express NaCT with a C-terminal Flag-tag (DYKDDDDK) that can be specifically and accurately recognized by commercially available anti-Flag antibodies. The 24-base Flag coding sequence was located immediately upstream of the natural translation termination codon (TAG) and was validated by Sanger sequencing. The general development, physical activities, and reproductive outcomes of this mouse strain were comparable to those of the C57BL/6 mice. No differences were detected between the Slc13a5Flag and wild-type mice. Tooth development was extensively characterized using dissection microscopy, bSEM, light microscopy, in situ hybridization, and immunohistochemistry. Tooth formation was not altered in any detectable way by the introduction of the Flag. The Slc13a5Flag citrate transporter was observed on all outer membranes of secretory ameloblasts (distal, lateral, and proximal), with the strongest signal on the Tomes process, and was detectable in all but the distal membrane of maturation-stage ameloblasts. The papillary layer also showed positive immunostaining for Flag. The outer membrane of odontoblasts stained stronger than ameloblasts, except for the odontoblastic processes, which did not immunostain. As NaCT is thought to only facilitate citrate entry into the cell, we performed in situ hybridization that showed Ank is not expressed by secretory- or maturation-stage ameloblasts, ruling out that ANK can transport citrate into enamel. In conclusion, we developed Slc13a5Flag reporter mice that provide specific and sensitive localization of a fully functional NaCT-Flag protein. The localization of the Slc13a5Flag citrate transporter throughout the ameloblast membrane suggests that either citrate enters enamel by a paracellular route or NaCT can transport citrate bidirectionally (into or out of ameloblasts) depending upon local conditions. Full article
(This article belongs to the Special Issue Molecular Metabolism of Ameloblasts in Tooth Development)
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9 pages, 429 KiB  
Article
The Problem of the Presence of DNA in Cosmetic and Medicinal Products Obtained from Animals on the CITES List
by Aleksandra Figura, Magdalena Gryzinska and Andrzej Jakubczak
Genes 2025, 16(7), 805; https://doi.org/10.3390/genes16070805 - 8 Jul 2025
Viewed by 298
Abstract
Background: The illegal trade in wildlife remains a critical threat to biodiversity, prompting the development of international regulatory frameworks such as the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). One of the key challenges in enforcement is [...] Read more.
Background: The illegal trade in wildlife remains a critical threat to biodiversity, prompting the development of international regulatory frameworks such as the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). One of the key challenges in enforcement is the detection of CITES-listed species in highly processed consumer goods. Methods: This study investigates the use of molecular techniques to detect animal DNA in two selected commercially available medicinal products—a balm and a gel—marketed with ingredients suggestive of protected species such as the brown bear (Ursus arctos) and the medicinal leech (Hirudo medicinalis). Results: Although DNA from these target species was not detected, the analysis revealed the presence of genetic material from the American mink (Neovison vison) and domestic pig (Sus scrofa), indicating the undeclared use of animal-derived substances. While limited in scope, these findings suggest potential ethical and transparency concerns, particularly for consumers adhering to vegetarian, vegan, or religious dietary practices. Conclusions: The study illustrates the feasibility of applying DNA-based screening methods in complex, degraded matrices and their potential as supportive tools in consumer product assessment. However, broader studies are necessary before drawing general regulatory or conservation conclusions. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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17 pages, 4748 KiB  
Article
Impact of the Gut Microbiota–Metabolite Axis on Intestinal Fatty Acid Absorption in Huainan Pigs
by Jing Wang, Liangying Zhu, Yangyang Wang, Qiang Ma, Xiangzhou Yan, Mingxun Li and Baosong Xing
Microorganisms 2025, 13(7), 1609; https://doi.org/10.3390/microorganisms13071609 - 8 Jul 2025
Viewed by 472
Abstract
The gut microbiota critically influences lipid metabolism and fat deposition in pigs, processes that underpin pork quality preferences and differentiate the meat traits of Chinese indigenous breeds (fat-type) from those of Western commercial breeds (lean-type). To explore the mechanisms underlying breed-specific fatty acid [...] Read more.
The gut microbiota critically influences lipid metabolism and fat deposition in pigs, processes that underpin pork quality preferences and differentiate the meat traits of Chinese indigenous breeds (fat-type) from those of Western commercial breeds (lean-type). To explore the mechanisms underlying breed-specific fatty acid absorption, we compared the rectal and colonic microbiota and metabolite profiles of Huainan and Large White pigs using 16S rRNA sequencing and untargeted metabolomics. HN pigs exhibited enriched Lactobacillus johnsonii and Lactobacillus amylovorus, along with a significantly higher Firmicutes/Bacteroidetes ratio. Functional predictions further revealed elevated microbial pathways related to glycolysis, pyruvate metabolism, and ABC transporters in HN pigs. Conversely, LW pigs showed increased abundance of potentially pro-inflammatory bacteria and enriched pathways for lipopolysaccharide (LPS) biosynthesis. Metabolites such as 4-ethyl-2-heptylthiazole and picolinic acid were significantly upregulated in HN pigs and served as robust biomarkers (Area Under the Curve, AUC = 1.0),with perfect discrimination observed in both rectal and colonic samples. Integrative analysis identified 52 co-enriched microbial and metabolic pathways in HN pigs, including short-chain fatty acid (SCFA) production, lipid biosynthesis and transport, amino acid metabolism, ABC transporter activity, and the PPAR signaling pathway, supporting a microbiota–metabolite axis that enhances fatty acid absorption and gut immune balance. These findings provide mechanistic insight into breed-specific fat deposition and offer candidate biomarkers for improving pork quality via precision nutrition and breeding. Full article
(This article belongs to the Section Veterinary Microbiology)
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17 pages, 2146 KiB  
Article
Development of an Effective Single-Dose PCV2/CSFV Bivalent Subunit Vaccine Against Classical Swine Fever Virus and Porcine Circovirus Type 2
by Yu-Chieh Chen, Wen-Bin Chung, Hso-Chi Chaung, Yen-Li Huang, Chi-Chih Chen and Guan-Ming Ke
Vaccines 2025, 13(7), 736; https://doi.org/10.3390/vaccines13070736 - 8 Jul 2025
Viewed by 563
Abstract
Background/Objectives: Porcine Circovirus Type 2 (PCV2) impairs pigs’ immune systems and increases susceptibility to co-infections, including Classical Swine Fever (CSF), a highly contagious disease listed by the World Organisation for Animal Health (WOAH) as notifiable. Therefore, swine operations in CSF-endemic regions are [...] Read more.
Background/Objectives: Porcine Circovirus Type 2 (PCV2) impairs pigs’ immune systems and increases susceptibility to co-infections, including Classical Swine Fever (CSF), a highly contagious disease listed by the World Organisation for Animal Health (WOAH) as notifiable. Therefore, swine operations in CSF-endemic regions are encouraged to immunize piglets with both PCV2 and CSFV vaccinations. Currently, there is no commercially available bivalent vaccine for PCV2/CSFV. Methods: In this study, a total of twenty 4-week-old SPF pigs were administered our formulated PCV2/CSFV bivalent subunit vaccine, containing soluble CSFV-E2 (50 µg) and PCV2-ORF2 (100 µg) antigens with a porcine-specific CpG adjuvant. After 4 weeks of vaccination, all pigs were evaluated for efficacy against PCV2 and CSFV. Results: Pigs were only immunized once and showed significantly increased neutralizing or ELISA antibody titers against both viruses four weeks post-vaccination. After viral challenges, vaccinated pigs displayed no clinical signs or lesions and had markedly reduced CSFV and PCV2 viral loads in the serum and tissues compared to controls. Conclusions: These results demonstrate that a single dose of the PCV2/CSFV bivalent subunit vaccine is safe and effective in young pigs, induces strong antibody responses, and suppresses viral replication, making it a promising tool for swine disease control and cost-effective vaccination strategies. Full article
(This article belongs to the Special Issue Vaccination Against Major Respiratory Pathogens in Livestock Farming)
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Article
Development and Application of a Multiplex Real-Time TaqMan qPCR Assay for the Simultaneous Detection of African Swine Fever Virus, Classical Swine Fever Virus, Porcine Reproductive and Respiratory Syndrome Virus, Pseudorabies Virus, and Porcine Circovirus Type 2
by Dongdong Yin, Shuangshuang Xu, Yayun Liu, Hao Guo, Mengdie Lan, Lei Yin, Jieru Wang, Yin Dai, Xuehuai Shen, Kai Zhan and Xiaocheng Pan
Microorganisms 2025, 13(7), 1573; https://doi.org/10.3390/microorganisms13071573 - 3 Jul 2025
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Abstract
Since its emergence in China in 2018, African swine fever virus (ASFV) has posed a severe threat to the pig farming industry due to its high transmissibility and mortality rate. The clinical signs of ASFV infection often overlap with those caused by other [...] Read more.
Since its emergence in China in 2018, African swine fever virus (ASFV) has posed a severe threat to the pig farming industry due to its high transmissibility and mortality rate. The clinical signs of ASFV infection often overlap with those caused by other swine viruses such as classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine circovirus type 2 (PCV2), making timely and precise diagnosis a considerable challenge. To address this, we established a TaqMan-based multiplex real-time quantitative PCR (qPCR) assay capable of simultaneously detecting ASFV, CSFV, PRRSV, PRV, and PCV2. Specific primer-probe sets were developed targeting conserved genomic regions: the ASFV P72 gene, CSFV 5’UTR region, PRRSV ORF6, PCV2 cap gene, and PRV gB gene. After thorough optimization, the assay demonstrated robust analytical performance, exhibiting strong target specificity with no cross-detection of non-target pathogens. The detection threshold was determined to be 10 copies/μL per virus, indicating high assay sensitivity. Repeatability analysis revealed low variability, with intra- and inter-assay coefficient of variation values remaining below 2.3%. When applied to 95 clinical samples, the multiplex assay yielded results that were fully consistent with those obtained using commercially available singleplex qPCR kits. In conclusion, the multiplex TaqMan qPCR method developed in this study is characterized by high specificity, sensitivity, and reproducibility. It provides a reliable and efficient diagnostic tool for the simultaneous detection and differential diagnosis of ASFV and other clinically similar viral infections in swine, thereby offering robust technical support for swine disease surveillance and control. Full article
(This article belongs to the Special Issue Viral Infection on Swine: Pathogenesis, Diagnosis and Control)
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