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1.
Biology (Basel) ; 13(6)2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38927250

RESUMEN

Artificial insemination (AI) with liquid-preserved semen has recently become common in pig breeding. The semen doses are produced in a centralized manner at the boar stud and then subsequently distributed and transported to sow farms. However, vibration emissions during transportation by logistic vehicles may adversely affect the quality of boar sperm. Therefore, this study aimed to explore the impact of vibration-induced emissions on sperm quality and function under simulated transportation conditions. Each time, ejaculates from all 15 boars were collected and then pooled together to minimize individual variations, and the sample was split using an extender for dilution. Different rotational speeds (0 rpm, 80 rpm, 140 rpm, 200 rpm) were utilized to simulate varying intensities of vibration exposure using an orbital shaker, considering different transportation times (0 h, 3 h, and 6 h). Subsequently, evaluations were conducted regarding sperm motility, plasma membrane integrity, acrosome integrity, mitochondrial function, adenosine triphosphate (ATP) levels, mitochondrial reactive oxygen species (ROS) levels, pH, glycolytic pathway enzyme activities, and capacitation following exposure to vibration emissions. Both vibration time and intensity impact sperm motility, plasma membrane integrity, and acrosomal integrity. Vibration exposure significantly reduced sperm ATP levels, mitochondrial membrane potential, and the levels of mitochondria-encoded proteins (MT-ND1, MT-ND6) (p < 0.05). After vibration emission treatment, the pH value and mitochondrial ROS levels significantly increased (p < 0.05). Inhibition of sperm glycolysis was observed, with reduced activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH), along with decreased lactate levels (p < 0.05). Additionally, sperm tyrosine phosphorylation levels were significantly reduced by vibration emissions compared to the control group (p < 0.05). After the vibration emission treatment, the number of sperm bound to each square millimeter of oviduct explants decreased significantly compared to the control group (p < 0.05). Similarly, compared to the control group, using semen subjected to vibration stress for AI results in significantly reduced pregnancy rates, total born litter size, live-born litter size, and healthy born litter size (p < 0.05).

2.
Viruses ; 15(6)2023 05 25.
Artículo en Inglés | MEDLINE | ID: mdl-37376538

RESUMEN

It is well established that PRRSV elimination is an effective strategy for PRRS control, but published reports concerning successful PRRSV elimination cases in farrow-to-finishing herds are rare. Here, we have reported a successful PRRSV elimination case in a farrow-to-finish herd by employing a "herd closure and rollover" approach with some modifications. Briefly, the introduction of pigs to the herd was stopped and normal production processes were maintained until the herd reached a PRRSV provisional negative status. During the herd closure, strict biosecurity protocols were implemented to prevent transmission between nursery pigs and sows. In the current case, introducing gilts before herd closure and live PRRSV exposure were skipped. In the 23rd week post-outbreak, the pre-weaning piglets started to show 100% PRRSV negativity in qPCR tests. In the 27th week, nursery and fattening barns fully launched depopulation. In the 28th week, nursery and fattening houses reopened and sentinel gilts were introduced into gestation barns. Sixty days post-sentinel gilt introduction, the sentinel pigs maintained being PRRSV antibody negative, manifesting that the herd matched the standard of the provisional negative status. The production performance of the herd took 5 months to bounce back to normal. Overall, the current study provided additional information for PRRSV elimination in farrow-to-finish pig herds.


Asunto(s)
Síndrome Respiratorio y de la Reproducción Porcina , Virus del Síndrome Respiratorio y Reproductivo Porcino , Enfermedades de los Porcinos , Porcinos , Animales , Femenino , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Síndrome Respiratorio y de la Reproducción Porcina/prevención & control , Sus scrofa , Destete
3.
BMC Vet Res ; 19(1): 2, 2023 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-36597091

RESUMEN

BACKGROUND: Porcine Teschovirus (PTV), also named Teschovirus A, is prevalent in pig populations, mainly causing neurological symptoms, diarrhea, pneumonia, and reproductive failure, however the morbidity and mortality are usually low in pig farms. CASE PRESENTATION: In this study, we reported a PTV outbreak investigation in one large-scale pig farm in China with severe symptoms including diarrhea, lethargy, locomotor ataxia, nystagmus, paralysis of the hind limbs, and coma in piglets. More importantly, the mortality reached 38% in suckling pigs, which is remarkably high in PTV history. A novel PTV strain, named HeNZ1, was isolated from cerebral samples of one suckling pig and the genome sequence was obtained by NGS sequencing. Phylogenetic and evolutionary divergence analyses revealed that HeNZ1 belongs to PTV genotype 2. Surprisingly, the VP1 coding region of HeNZ1 shares the highest sequence similarity with European PTV-2 strains, instead of China domestic PTV-2 strains, implying it may not derive from China local PTV-2 strains. Multiple sequence alignment and B cell epitope prediction of PTV VP1 and VP2 protein revealed 10 B cell epitopes, 5 mutant clusters and 36 unique mutation sites, of which 19 unique mutation sites are located in B cell epitopes and exposed on the surface of VP1 or VP2, implying significant antigenic drift potential of HeNZ1. CONCLUSION: These results indicate that HeNZ1 is a highly virulent PTV-2 strain, which capable of causing severe neurological symptoms and high mortality in piglets. Bioinformatic analysis suggest that HeNZ1 is genetically and antigenically different from other Chinese PTV-2 strains. Overall, current case expanded our understanding of PTV-2 clinical spectrum and revealed the emergence of a highly virulent PTV-2 strain with substantial genetic diversity and antigenic drift potential in VP1 and VP2.


Asunto(s)
Encefalomielitis , Infecciones por Picornaviridae , Enfermedades de los Porcinos , Teschovirus , Porcinos , Animales , Filogenia , Epítopos de Linfocito B , Diarrea/veterinaria , China/epidemiología , Encefalomielitis/veterinaria , Infecciones por Picornaviridae/veterinaria
4.
Meat Sci ; 196: 109052, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36455423

RESUMEN

Accurate and rapid determination of meat quality traits plays key roles in food industry and pig breeding. Currently, most of the spectroscopic instruments developed for meat quality determination can only obtain the spectral average value of the sample, so it is difficult to evaluate the spatial variation of meat quality traits. In this study, we evaluated the predictive potential of 14 meat quality traits based on large-scale VIS/NIR hyperspectral images collected by SpecimIQ. When predictions were based solely on hyperspectral data, the prediction accuracy (R2cv) for the majority of meat qualities ranged from 0.60 to 0.70. After adding texture information, the prediction accuracy of all traits is improved by different magnitudes (R2cv increases from 1.5% to 16.4%). Finally, the best model was utilized to visualize the spatial distribution of Fat (%) and Moisture (%) to assess their homogeneity. These results suggest that hyperspectral imaging has great potential for predicting and visualizing various meat qualities, as well as industrial applications for automated measurements.


Asunto(s)
Carne de Cerdo , Carne Roja , Porcinos , Animales , Imágenes Hiperespectrales/veterinaria , Carne , Fenotipo
5.
Metabolites ; 12(12)2022 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-36557235

RESUMEN

Excessive fat accumulation is a common phenomenon in cultured fish, which can cause metabolic disease such as fatty liver. However, the relative regulatory approach remains to be explored. Based on this, two feeding trials were conducted. Firstly, fish were fed either a normal-fat diet (NFD) or a high-fat diet (HFD) for eight weeks and sampled at the 2nd, 4th, 6th, and 8th week after feeding (Experiment I). In the first four weeks, fish fed an HFD grew faster than those fed an NFD. Conversely, the body weight and weight gain were higher in the NFD group at the 6th and 8th weeks. Under light and transmission electron microscopes, fat accumulation of the liver was accompanied by an obvious endoplasmic reticulum (ER) swell. Accordingly, the expressions of atf-6, ire-1, perk, eif-2α, atf-4, grp78, and chop showed that ER stress was activated at the 6th and 8th weeks. In Experiment II, 50 mg/kg 4-PBA (an ERs inhibitor) was supplemented to an HFD; this was named the 4-PBA group. Then, fish was fed with an NFD, an HFD, and a 4-PBA diet for eight weeks. As the result, the excessive fat deposition caused by an HFD was reversed by 4-PBA. The expression of ER stress-related proteins CHOP and GRP78 was down-regulated by 4-PBA, and the transmission electron microscope images also showed that 4-PBA alleviated ER stress induced by the feeding of an HFD. Furthermore, 4-PBA administration down-regulated SREBP-1C/ACC/FAS, the critical pathways of fat synthesis. In conclusion, the results confirmed that ER stress plays a contributor role in the fat deposition by activating the SREBP-1C/ACC/FAS pathway. 4-PBA as an ER stress inhibitor could reduce fat deposition caused by an HFD via regulating ER stress.

6.
Int J Biol Macromol ; 222(Pt B): 1778-1788, 2022 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-36195228

RESUMEN

p-Coumaric acid and caffeic acid were grafted onto chitosan through a non-radical synthesis method to improve the properties of chitosan and expand its application in food industry. Structural characterization demonstrated that the -COOH of the two phenolic acids were bonded to the -NH2 of the chitosan and formed an acylamino. The grafting ratios of p-coumaric acid-modified chitosan (Cm-CTS) and caffeic acid-modified chitosan (Cf-CTS) reached 10.30 % and 9.78 %, respectively. After modification, the water solubility of the chitosan greatly improved from 9.33 % (native chitosan, Nt-CTS) to 77.33 % (Cm-CTS) and 100 % (Cf-CTS). Besides, the involvement of phenolic acid and caffeic acid endowed the chitosan with strengthened antioxidation and antibacterial activities against Escherichia coli and Staphylococcus aureus. Nt-CTS and the modified chitosans were coated on the pork surface. The results indicated that Nt-CTS effectively inhibited pork spoilage and the modified chitosans could further prolong the shelf life of pork.


Asunto(s)
Quitosano , Carne de Cerdo , Carne Roja , Animales , Porcinos , Quitosano/química , Amidas/farmacología , Ácidos Cafeicos/química , Antibacterianos/farmacología , Escherichia coli
7.
Antioxidants (Basel) ; 11(7)2022 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-35883767

RESUMEN

Oxidative stress is a common phenomenon in aquaculture, which can be induced by nutritional or environmental factors. Generally, oxidative stress causes poor growth performance, metabolic dysregulation, and even the death of aquatic animals. To identify a nutritional intervention strategy, high-fat diet (HFD) feeding (Experiment I) and acute ammonia nitrogen challenge (Experiment II) tests were carried out. In Experiment I, HFD feeding significantly decreased the growth performance concomitantly with excessive fat deposition in the liver and abdomen. The addition of 4-PBA in the diet improved the excessive fat accumulation. The activities of antioxidative enzymes were suppressed, and the levels of lipid and protein peroxidation were increased, indicating that HFD feeding induced oxidative stress. The endoplasmic reticulum stress (ERs) related genes were downregulated in the HFD group. Under a transmission electron microscope (TEM), more swollen and dilated ER lumen could be observed. These results indicated that the HFD induced ERs activation. Although 4-PBA acted as a potent ERs inhibitor, as evidenced by the alleviated alterations of ERs molecules and the ER ultrastructure, the oxidative stress was also attenuated by 4-PBA. In Experiment II, dietary 4-PBA improved the tolerance to the acute ammonia nitrogen challenge, as lower mortality and serum aminotransferase activity was found. Further results showed that 4-PBA decreased the peroxidation content and attenuated ERs, thus confirming the correlation between oxidative stress and ERs. Our findings showed that dietary 4-PBA supplementation can attenuate oxidative stress induced by a HFD or acute ammonia challenge; the mechanism is related to its potent inhibition effect for ERs.

8.
J Hazard Mater ; 431: 128498, 2022 06 05.
Artículo en Inglés | MEDLINE | ID: mdl-35278944

RESUMEN

To prevent the toxic effect of amantadine (AMD) on humans, a sensitive and direct detection method is required. The conventional enzyme-linked immunosorbent assay (ELISA) usually shows a monochromatic gradient color variation with the concentration of the target; hence, it is not a sensitive method for naked-eye detection. In this work, Ag-Au nanorings with highly tunable plasmon properties were synthesized as colorimetric nanosensors. The growth of Ag on the hollow nanorings led to rich color variations. Ag-Au nanorings were integrated into ELISA for the sensitive detection of AMD with the naked eye. The proposed method showed high sensitivity for the qualitative and quantitative detection of AMD, the visible cut-off value (0.2 ng mL-1) and limit of detection (0.071 ng mL-1) were 10-fold and 4.7-fold lower, respectively, than those of conventional ELISA. This method showed a linear range of 0.08-2 ng mL-1 and 4-12 ng mL-1. The detection results of this method on 100 samples (food samples and municipal water) agreed well with those of liquid chromatography-tandem mass spectrometry. The proposed plasmonic ELISA has high sensitivity, easy operation, and naked-eye readout.


Asunto(s)
Oro , Nanopartículas del Metal , Amantadina/análisis , Colorimetría , Ensayo de Inmunoadsorción Enzimática , Oro/química , Humanos , Límite de Detección , Nanopartículas del Metal/química
9.
Anim Nutr ; 8(1): 265-276, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34988308

RESUMEN

This study aimed to assess the changes of small intestinal morphology, progenitors, differentiated epithelial cells, and potential mechanisms in neonatal piglets. Hematoxylin and eosin staining of samples from 36 piglets suggested that dramatic changes were observed in the jejunum crypts depth and crypt fission index of neonatal piglets (P < 0.001). The number of intestinal stem cells (ISC) tended to increase (P < 0.10), and a decreased number of enteroendocrine cells appeared in the jejunal crypt on d 7 (P < 0.05). Furthermore, the mRNA expression of jejunal chromogranin A (ChgA) was down-regulated in d 7 piglets (P < 0.05). There was an up-regulation of the adult ISC marker gene of SPARC related modular calcium binding 2 (Smoc2), and Wnt/ß-catenin target genes on d 7 (P < 0.05). These results were further verified in vitro enteroid culture experiments. A mass of hollow spheroids was cultured from the fetal intestine of 0-d-old piglets (P < 0.001), whereas substantial organoids with budding and branching structures were cultured from the intestine of 7-d-old piglets (P < 0.001). The difference was reflected by the organoid budding efficiency, crypt domains per organoid, and the surface area of the organoid. Furthermore, spheroids on d 0 had more Ki67-positive cells and enteroendocrine cells (P < 0.05) and showed a decreasing trend in the ISC and goblet cells (P < 0.10). Moreover, the mRNA expression of spheroids differed markedly from that of organoids, with low expression of intestinal differentiation gene (Lysozyme; P < 0.05), epithelial-specific markers (Villin, E-cadherin; P < 0.05), and adult ISC markers (leucine-rich repeat-containing G protein-coupled receptor 5 [Lgr5], Smoc2; P < 0.001), and up-regulation of fetal marker (connexin 43 [Cnx43]; P < 0.05). The mRNA expression of relevant genes was up-regulated, and involved in Wnt/ß-catenin, epidermal growth factor (EGF), Notch, and bone morphogenetic protein (BMP) signaling on d 7 organoids (P < 0.05). Spheroids displayed low differentiated phenotype and high proliferation, while organoids exhibited strong differentiation potential. These results indicated that the conversion from the fetal progenitors (spheroids) to adult ISC (normal organoids) might largely be responsible for the fast development of intestinal epithelial cells in neonatal piglets.

10.
J Hazard Mater ; 423(Pt B): 127204, 2022 02 05.
Artículo en Inglés | MEDLINE | ID: mdl-34555767

RESUMEN

Herein, we propose a lateral flow immunoassay (LFIA) based on the dual spectral-overlapped fluorescence quenching of polydopamine nanospheres (PDANs) caused by the inner filter effect to sensitively detect sulfamethazine (SMZ). The fluorescence quenching LFIA device consists of four parts: absorbent pad, polyvinyl chloride pad, sample pad, and nitrocellulose membrane. Compared with traditional quenchers such as gold nanoparticles (AuNPs) with single spectral-overlapped quenching ability, PDANs can quench the excitation light and emission light of three fluorescence donors (aggregation-induced emission fluorescent microsphere, AIEFM; fluorescent microsphere, FM; and quantum dot bead, QB). The fluorescence intensity changes (ΔF) are numerically larger for PDANs-LFIA (ΔFAIEFM = 2315, ΔFFM = 979, ΔFQB = 910) than those for AuNPs-LFIA (ΔFAIEFM = 1722, ΔFFM = 833, ΔFQB =;520). AIEFM-based PDANs-LFIA exhibits a large ΔF (2315) in response to the changes in the SMZ concentration, and produces a high signal-to-noise ratio. The limit of detection (LOD) and visual LOD of LFIA based on PDANs quenching AIEFM for the detection of SMZ in chicken are 0.043 and 0.5 ng/mL, respectively. The results confirm that the proposed method can be used for the detection of hazardous materials in practical applications.


Asunto(s)
Nanopartículas del Metal , Nanosferas , Oro , Inmunoensayo , Indoles , Límite de Detección , Polímeros , Sulfametazina
11.
Int J Biol Macromol ; 194: 246-253, 2022 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-34875310

RESUMEN

To further improve the performance of chitosan in food processing and preservation, this study investigated the grafting of the caffeic acid onto the chitosan in non-enzymatic and enzymatic systems. Result suggested that the caffeic acid was successfully incorporated into the chitosan in the non-enzymatic system, and the grafting ratio of modified chitosan (CA@CTS-N) was 7.49%. Moreover, lipase had a significant positive effect on the grafting reaction of the chitosan, and the modified chitosan prepared in enzymatic system (CA@CTS-E) obtained a higher grafting ratio, which was 11.82%. In both systems, the carboxyl of the caffeic acid was bonded to the amino of the chitosan and formed carbonyl ammonia. After the introduction of foreign group, many changes occurred in the functional properties of the modified chitosan. First, the water solubility of the chitosan was significantly improved from 0.00285 (native chitosan, CTS) to 0.221 (CA@CTS-N) and 0.774 g/100 mL (CA@CTS-E). The caffeoyl had a significant impact on the emulsifying properties of the chitosan. Compared with those of CTS, the modified chitosan had stronger antioxidation and antibacterial activities against Escherichia coli, Staphylococcus aureus, and Candida albicans. Finally, the pork treated with the modified chitosan exhibited longer shelf life than that treated with CTS.


Asunto(s)
Ácidos Cafeicos/química , Quitosano/análisis , Quitosano/química , Carne de Cerdo , Preservación Biológica , Fenómenos Químicos , Concentración de Iones de Hidrógeno , Carne Roja , Solubilidad , Análisis Espectral
12.
Viruses ; 13(12)2021 12 16.
Artículo en Inglés | MEDLINE | ID: mdl-34960796

RESUMEN

Porcine epidemic diarrhea virus (PEDV), an enteric coronavirus, causes neonatal pig acute gastrointestinal infection with a characterization of severe diarrhea, vomiting, high morbidity, and high mortality, resulting in tremendous damages to the swine industry. Neither specific antiviral drugs nor effective vaccines are available, posing a high priority to screen antiviral drugs. The aim of this study is to investigate anti-PEDV effects of carbazole alkaloid derivatives. Eighteen carbazole derivatives (No.1 to No.18) were synthesized, and No.5, No.7, and No.18 were identified to markedly reduce the replication of enhanced green fluorescent protein (EGFP) inserted-PEDV, and the mRNA level of PEDV N. Flow cytometry assay, coupled with CCK8 assay, confirmed No.7 and No.18 carbazole derivatives displayed high inhibition effects with low cell toxicity. Furthermore, time course analysis indicated No.7 and No.18 carbazole derivatives exerted inhibition at the early stage of the viral life cycle. Collectively, the analysis underlines the benefit of carbazole derivatives as potential inhibitors of PEDV, and provides candidates for the development of novel therapeutic agents.


Asunto(s)
Antivirales/farmacología , Carbazoles/farmacología , Virus de la Diarrea Epidémica Porcina/efectos de los fármacos , Animales , Antivirales/química , Carbazoles/química , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Relación Dosis-Respuesta a Droga , Estructura Molecular , Células Vero , Acoplamiento Viral/efectos de los fármacos , Replicación Viral/efectos de los fármacos
13.
Anim Nutr ; 7(4): 1070-1077, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34738037

RESUMEN

The present study aimed to investigate whether inflammation-associated responses in piglets are induced by high protein (HP) through activating nuclear factor kappa B (NF-κB) signaling. Sixteen piglets (35 d of age, Duroc × [Landrace × Yorkshire], weaned at d 21, initial BW = 9.70 ± 0.11 kg) were allocated to 18% and 26% CP (HP group) at random, comprising 8 replicate pens per treatment. The piglets were slaughtered to collect intestinal tissues when apparent, persistent, and stable diarrhea syndromes happened (on d 12). No significant differences were observed in their growth performance (P > 0.05), but reduction by 19.11%, 25.31%, 23.64% of ADFI, ADG, and G:F, respectively was detected in the HP group. The HP group had greater (P = 0.002) diarrhea rates. Furthermore, dietary HP had lower ileal villus height (VH; P = 0.048), ratio of villus height to crypt depth (VH/CD ratio; P = 0.016), and colonic CD (P = 0.034), as well as had the trend (P = 0.075) to reduce the ileal villus absorptive area. Moreover, HP diets significantly elevated the goblet cell numbers in the ileal villi (P = 0.016) and colonic crypts (P < 0.001) and up-regulated (P = 0.012) the mRNA expression of mucin2 (Muc2) in the ileum. In addition, HP diets increased the myeloperoxidase concentration in the ileum (P = 0.002) and colon (P = 0.007) of piglets. Dietary HP significantly down-regulated the mRNA expression of tumor necrosis factor-α (TNF-α; P < 0.001) in the ileum, induced nitric oxide synthase (iNOS; P = 0.040) and interleukin-22 (IL-22; P = 0.008) in the colon, and inclined to down-regulate interleukin-1ß (IL-1ß; P = 0.076) expression in the colon. The relative protein abundance of Galectin-3 (P = 0.046) in the colon and the ratio of phosphorylation NF-κB to NF-κB (p-NF-κB/NF-κB ratio) in the ileum of HP piglets were also greater (P = 0.038). These results suggest that dietary HP may cause diarrhea in piglets by activating NF-κB signaling induced intestinal inflammation.

14.
Int J Biol Macromol ; 190: 343-350, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34492247

RESUMEN

In this paper, a method for the enzymatic modification of pectin, in which gallic acid (GA) and propyl gallate (PG) were grafted onto pectin molecules in an aqueous/organic two-phase system catalyzed by lipase, was proposed. The potential reaction mechanism was explored through UV-Vis, FTIR and 1H NMR spectroscopic methods and density functional theory. Results suggested that the lipase played a dual role during the modification by catalyzing the hydrolysis of methyl ester bonds of pectin in the aqueous phase and the esterification between the 4-OH of GA and PG and the -COOH of pectin in the organic phase. Moreover, the effects of GA and PG on the antioxidant and the antibacterial activities of pectin were evaluated, and results showed that the antioxidant and the antibacterial activities of modified pectin were better than those of native pectin. The effect of modified pectin on the quality of fresh bass (Lateolabrax maculatus) was further studied. Results suggested that, compared to control group, the total viable count, histamine level, malondialdehyde content and acid value of bass fillets treated with modified pectin were significantly reduced, whereas the sensory score was significantly increased.


Asunto(s)
Antibacterianos/farmacología , Antioxidantes/farmacología , Ácido Gálico/química , Pectinas/química , Galato de Propilo/química , Animales , Lubina , Teoría Funcional de la Densidad , Escherichia coli/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Conformación Molecular , Espectroscopía de Protones por Resonancia Magnética , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/efectos de los fármacos
15.
Viruses ; 13(7)2021 07 08.
Artículo en Inglés | MEDLINE | ID: mdl-34372529

RESUMEN

Pseudorabies virus (PRV) is an economically significant swine infectious agent. A PRV outbreak took place in China in 2011 with novel virulent variants. Although the association of viral genomic variability with pathogenicity is not fully confirmed, the knowledge concerning PRV genomic diversity and evolution is still limited. Here, we sequenced 54 genomes of novel PRV variants isolated in China from 2012 to 2017. Phylogenetic analysis revealed that China strains and US/Europe strains were classified into two separate genotypes. PRV strains isolated from 2012 to 2017 in China are highly related to each other and genetically close to classic China strains such as Ea, Fa, and SC. RDP analysis revealed 23 recombination events within novel PRV variants, indicating that recombination contributes significantly to the viral evolution. The selection pressure analysis indicated that most ORFs were under evolutionary constraint, and 19 amino acid residue sites in 15 ORFs were identified under positive selection. Additionally, 37 unique mutations were identified in 19 ORFs, which distinguish the novel variants from classic strains. Overall, our study suggested that novel PRV variants might evolve from classical PRV strains through point mutation and recombination mechanisms.


Asunto(s)
Genoma Viral , Herpesvirus Suido 1/genética , Filogenia , Seudorrabia/epidemiología , Análisis de Secuencia de ADN , Animales , Brotes de Enfermedades , Variación Genética , Genómica , Genotipo , Herpesvirus Suido 1/clasificación , Sistemas de Lectura Abierta , Seudorrabia/virología , Recombinación Genética , Porcinos , Enfermedades de los Porcinos/virología
16.
Monoclon Antib Immunodiagn Immunother ; 39(6): 222-227, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33351712

RESUMEN

Porcine epidemic diarrhea virus (PEDV) is an enteric swine coronavirus. Recent PEDV eruption in East Asian and North American countries made it notorious and caused huge economic losses to the porcine industry. Nonstructural protein 13 (nsp13) is a nucleic acid helicase/NTPase that plays a critical role in viral gene transcription and viral replication. To investigate the function of nsp13 in the context of PEDV infection, here, PEDV nsp13 gene was amplified and cloned into pET28a/pET30a/pGEX-6P-1 expression vectors. The recombinant his-tagged nsp13 and GST-tagged nsp13 were expressed. Purified his-tagged nsp13 from pET28a-nsp13 vectors was chosen for immunization. Three monoclonal antibodies (mAbs) named 5A9, 5C7, and 5G7 were identified from the hybridoma cells, and were characterized by Western blot analysis and immunofluorescent assay, which demonstrated high specificity of these three mAbs with pCAGGS-HA-nsp13. All three mAbs belong to IgG1+ kappa subclass. However, only mAb 5A9 could effectively and specifically recognize PEDV expressing nsp13. Furthermore, the generated antibody against nsp13 could be applied to investigate nsp13 function during PEDV replication.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Gastroenteritis Porcina Transmisible/terapia , Virus de la Diarrea Epidémica Porcina/inmunología , Proteínas no Estructurales Virales/inmunología , Animales , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/uso terapéutico , Línea Celular , Femenino , Células HEK293 , Humanos , Inmunización , Metiltransferasas/inmunología , Ratones , Ratones Endogámicos BALB C , ARN Helicasas/inmunología , Porcinos
17.
Virus Genes ; 56(6): 687-695, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32944812

RESUMEN

Porcine deltacoronavirus (PDCoV) has been recently identified as an emerging enteropathogenic coronavirus that mainly infects newborn piglets and causes enteritis, diarrhea and high mortality. Although coronavirus N proteins have multifarious activities, the subcellular localization of the PDCoV N protein is still unknown. Here, we produced mouse monoclonal antibodies against the PDCoV N protein. Experiments using anti-haemagglutinin antibodies and these monoclonal antibodies revealed that the PDCoV N protein is shuttled into the nucleolus in both ectopic PDCoV N-expressing cells and PDCoV-infected cells. The results of deletion mutagenesis experiments demonstrated that the predicted nucleolar localization signal at amino acids 295-318 is critical for nucleolar localization. Cumulatively, our study yielded a monoclonal antibody against the PDCoV N protein and revealed a mechanism by which the PDCoV N protein translocated into the nucleolus. The tolls and findings from this work will facilitate further investigations on the functions of the PDCoV N protein.


Asunto(s)
Nucléolo Celular/genética , Infecciones por Coronavirus/virología , Proteínas de la Nucleocápside de Coronavirus/genética , Deltacoronavirus/genética , Gastroenteritis Porcina Transmisible/virología , Interacciones Huésped-Patógeno/genética , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/química , Anticuerpos Antivirales/biosíntesis , Anticuerpos Antivirales/química , Línea Celular , Nucléolo Celular/metabolismo , Infecciones por Coronavirus/patología , Proteínas de la Nucleocápside de Coronavirus/metabolismo , Deltacoronavirus/crecimiento & desarrollo , Deltacoronavirus/metabolismo , Células Epiteliales/metabolismo , Células Epiteliales/ultraestructura , Células Epiteliales/virología , Gastroenteritis Porcina Transmisible/patología , Expresión Génica , Hemaglutininas Virales/genética , Hemaglutininas Virales/metabolismo , Riñón/patología , Riñón/virología , Ratones , Señales de Localización Nuclear , Transporte de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Eliminación de Secuencia , Porcinos
18.
J Colloid Interface Sci ; 578: 124-134, 2020 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-32521352

RESUMEN

Pollution of tetracyclines (TCs) in swine wastewater has been a critical concern worldwide. Notably, multiple anions (e.g. PO43-, NO3-) coexist in the actual environments, which could significantly influence the TCs removal. In the current study, MgAl layered double oxide (MgAl-LDO) was adopted for investigating the TC removal performance with/without PO43- or NO3-. In all systems, the adsorption performance exhibited two different approaches between low and high TC concentrations. In the single system, pseudo-second-order and the Freundlich model fitted well to the equilibrium adsorption data when TC concentration was below 125 mg·L-1, while the pseudo-first-order and the linear model could describe the removal process at high TC concentration (>125 mg·L-1). The maximum adsorption capacity was 83.56 mg·g-1. In the co-existing system, the adsorption capacity was slightly enhanced when TC concentration below 150 mg·L-1 however was inhibited at high concentration (>150 mg·L-1). Combined with the characterization analyses, the interaction mechanism at low concentration was primarily surface adsorption on reconstructed LDH from LDO in the TC-alone system. It is worth mention that both PO43- and NO3- facilitated the formation of LDH via rehydration of LDO which enhanced surface adsorption in the co-existing system. At high TC concentration, the formation of tetracycline-metal complexes played a dominant role in TC removal in the single system, whereas diminished complexation in the binary system led to the decreased TC removal. This study provides a theoretical and practical guidance for MgAl-LDO on the efficient remediation of actual tetracyclines wastewater.


Asunto(s)
Óxidos , Contaminantes Químicos del Agua , Adsorción , Animales , Cinética , Nitratos , Fosfatos , Porcinos , Tetraciclinas
19.
Asian-Australas J Anim Sci ; 32(12): 1809-1815, 2019 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-30744341

RESUMEN

OBJECTIVE: Copy number variations (CNVs) are a major source of genetic diversity complementary to single nucleotide polymorphism (SNP) in animals. The aim of the study was to performed a comprehensive genomic analysis of CNVs based on high density whole-genome SNP markers in Chinese Dongxiang spotted pigs. METHODS: We used customized Affymetrix Axiom Pig1.4M array plates containing 1.4 million SNPs and the PennCNV algorithm to identify porcine CNVs on autosomes in Chinese Dongxiang spotted pigs. Then, the next generation sequence data was used to confirm the detected CNVs. Next, functional analysis was performed for gene contents in copy number variation regions (CNVRs). In addition, we compared the identified CNVRs with those reported ones and quantitative trait loci (QTL) in the pig QTL database. RESULTS: We identified 871 putative CNVs belonging to 2,221 CNVRs on 17 autosomes. We further discarded CNVRs that were detected only in one individual, leaving us 166 CNVRs in total. The 166 CNVRs ranged from 2.89 kb to 617.53 kb with a mean value of 93.65 kb and a genome coverage of 15.55 Mb, corresponding to 0.58% of the pig genome. A total of 119 (71.69%) of the identified CNVRs were confirmed by next generation sequence data. Moreover, functional annotation showed that these CNVRs are involved in a variety of molecular functions. More than half (56.63%) of the CNVRs (n = 94) have been reported in previous studies, while 72 CNVRs are reported for the first time. In addition, 162 (97.59%) CNVRs were found to overlap with 2,765 previously reported QTLs affecting 378 phenotypic traits. CONCLUSION: The findings improve the catalog of pig CNVs and provide insights and novel molecular markers for further genetic analyses of Chinese indigenous pigs.

20.
Anim Sci J ; 89(6): 868-875, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29602194

RESUMEN

Pigs show extensive variation in exterior appearance. This variation has been explored as one of the selection targets to form breeding features in the pig industry. In this study, a customized Affymetrix Axiom Pig1.4M array plate was used to conduct genome-wide association studies (GWAS) for two exterior traits-spotted coat color (SC) and facial type (FT)-in Chinese Dongxiang spotted pigs. Two and 12 single nucleotide polymorphisms (SNPs) were identified to be associated with SC and FT at the 5% genome-wide significant level, respectively. Of these SNPs, two associated with SC were located around the DACH1 gene on Sus Scrofa chromosome (SSC)11. Eleven SNPs associated with FT were located within a 7.68 Mb region (29.03-36.71 Mb) on SSC7. Based on the GWAS results and the biological functions of genes, we highlight EDNRB as a candidate gene for SC and HMGA1 and RPS10 as the potential genes affecting facial variation. The findings contribute to the final characterization of causative genes and mutations underlying the effects of the SSC7 and SSC11 loci and improve our understanding of the genetic basis of phenotypic variation in Chinese indigenous pigs.


Asunto(s)
Variación Genética/genética , Estudio de Asociación del Genoma Completo/métodos , Fenotipo , Carácter Cuantitativo Heredable , Porcinos/genética , Animales , Cromosomas/genética , Color , Proteínas del Ojo/genética , Femenino , Proteína HMGA1a/genética , Cabello , Humanos , Masculino , Polimorfismo de Nucleótido Simple , Receptor de Endotelina B/genética , Proteínas Ribosómicas/genética , Factores de Transcripción/genética
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