BACKGROUND:Bovine bone high-temperature hydrolysate (BBHH) is the main raw material that is used to prepare emulsified products including sauce packets, beef bone broth and bovine bone white soup. These traditional foods are favored by consumers for their unique nutritional value and sensory quality in Asia. However, as an emulsifier, BBHH alone has limited emulsification and stabilization capacity. RESULTS:This study investigated how NaCl affects the stability of BBHH emulsion by influencing the adsorption properties, microstructure and physicochemical interactions. The adsorption rate (34.65 ± 2.05%) and thickness of the adsorbed layer (100.55 ± 0.45 nm) were the highest at 100 mm NaCl. The cross-linking of oil droplets was enhanced at 100 mm NaCl, whereas the fine network of emulsion was damaged with excessive NaCl concentration (200 mm) observed by cryo-scanning electron microscopy. Raman spectroscopy confirmed that the unfolding of the interfacial protein and strengthening oil-protein interactions increased emulsification. Moreover, BBHH emulsions showed different emulsifying stability by NaCl induction, such that the emulsifying stability index values were 78.09 ± 1.21% at 100 mm NaCl and 68.91 ± 0.35% at 200 mm, which could be elucidated by the interfacial properties, indicating that the interfacial properties were essential for the emulsions. CONCLUSION:The present study demonstrated that BBHH emulsion exhibited good stability at lower ionic strength (≤ 100 mm NaCl). These discoveries provide valuable insights into the relationship between emulsion stability and interfacial properties, as well as contribute to a further understanding of NaCl induction in the food industry. © 2025 Society of Chemical Industry.

Abstract The coronavirus porcine epidemic diarrhea virus (PEDV) is still widespread in pigs rapidly due to its highly contagious, resulting in huge economic losses to the pig industry. Herein, we investigated the molecular divergence between PEDV and other related coronaviruses, the results showed that the spike (S) gene exhibited larger dS (synonymous substitutions per synonymous site) values than other genes. In the assay of the extent of positive selection, eight amino acid (aa) sites of S protein showed strong signals of positive selection, and seven aa sites of them were located in the surface of S protein (S1 domain), suggesting the high selection pressure of S protein during receptor binding. We analyzed the complete genomes of 647 strains retrieved from the GenBank database. Topologically, the high similarity between the complete genome and S gene indicated that the S gene is more representative of the evolutionary relationship at the genome-wide level than other genes. Structurally, the dominant of the highly mutated residues of S protein indicated that the evolutionary pattern is highly S1 domain related. We constructed the haplotype networks using the PEDV S gene, the results showed that the strains are obviously clustered geographically in the lineages corresponding to genotype GI and GII. The alignment analysis on representative strains in the main haplotypes revealed 3 distinguishable nucleic acid sites among those strains, suggesting a putative evolutionary mechanism of PEDV. These findings provide several new fundamental insights into the evolution of PEDV and the guidance for developing effective prevention countermeasures against PEDV.

Immunoglobulin A (IgA) of sows is critically important for assessing piglets' protective capacity against porcine epidemic diarrhea virus (PEDV). Here, we report a therapeutic chimeric anti-PEDV IgG/IgA expressed by Chinese hamster ovary (CHO) cells for oral treatment of PED. The chimeric anti-PEDV IgG/IgA was produced by the CHO cell lines, in which the heavy chain was constructed by combining the VH, Cγ1 and hinge regions of PEDV IgG mAb 8A3, and the Cα2 and Cα3 domains of a Mus musculus immunoglobulin alpha chain. The chimeric anti-PEDV IgG/IgA could neutralize the strains of CV777 (G1), P014 (G2) and HN1303 (G2) in vitro effectively, showing broad-spectrum neutralization activity. The in vivo challenge experiments demonstrated that chimeric anti-PEDV IgG/IgA (9C4) produced in the CHO cell supernatant could alleviate clinical diarrhea symptoms of the PEDV infection in piglets. In general, our study showed that chimeric anti-PEDV IgG/IgA produced from CHO cell line supernatants effectively alleviates PEDV infection in piglets, which also gives the foundation for the construction of fully functional secretory IgA by the J chain introduction to maximize the antibody therapeutic effect.

The large-scale outbreaks of Pseudorabies in china since 2011 in vaccine-immunized pig farms were caused by the highly virulent Pseudorabies virus (PRV). To investigate the factors involved in the virulence enhancement of the variant PRV, four recombinant viruses with the virulence gene’s replacement, gI/gE/11K and TK/gI/gE/11K , between the variant strain HN1201 and the classical strain Fa were constructed based on bacterial artificial chromosome infectious clones. Compared with their parental strain, the viral titers of the recombinant PRV strains are not strongly influenced by the replacement of TK/gI/gE/11K , while as previously reported, the strain HN1201 and its derived viruses showed the higher mortality, the severer clinical symptoms and tissues damage than that of strain Fa and its derived strains. In summary, the TK/gI/gE/11K genes of variant strain HN1201 showed no contribute to its virulence enhancement compares with the classical strain.

我国生猪产业发展中,重大疫病的流行可造成产业巨大经济损失,包括2006年暴发的高致病性猪蓝耳病、2011年底开始的由猪伪狂犬病病毒变异株引起的猪伪狂犬病,特别是自2018年8月以来,由非洲猪瘟病毒感染引起的非洲猪瘟疫情使我国养猪业遭受重创.非洲猪瘟病毒传播途径多样,在环境中可长时间存活,难以清除,且目前无疫苗用于预防,因此,非洲猪瘟病毒将与其他疫病一起继续威胁我国生猪产业的发展.

自2019年12月以来,由SARS-CoV-2感染造成的COVID-19对全球人类的健康构成了巨大威胁.为研究SARS-CoV-2与猪冠状病毒之间的相关性,本实验采用生物信息学分析方法对SARS-CoV-2和常见猪冠状病毒的纤突(S)蛋白氨基酸序列进行遗传进化分析、对S蛋白S1亚基的受体结合域空间结构进行相似性分析,并采用双抗原夹心ELISA试剂盒检测了常见猪冠状病毒阳性血清中的SARS-CoV-2抗体.结果显示,SARS-CoV-2与猪冠状病毒的遗传关系较远,其S1亚基的受体结合域空间结构差异大,常见猪冠状病毒阳性血清中的SARS-CoV-2特异性抗体均为阴性.表明SARS-CoV-2与猪冠状病毒亲缘关系远,难以通过感染猪进而接触人类以实现跨物种感染,威胁公共卫生安全的风险极小.本研究为COVID-19病原溯源与跨物种生物安全性分析提供基础.

Foot-and-mouth disease (FMD), caused by FMD virus (FMDV), is a highly contagious and economically devastating viral disease of cloven-hoofed animals worldwide. In this study, the coexpression of small ubiquitin-like modifier (SUMO)–fused capsid proteins of FMDV serotype O by single plasmid in Escherichia coli was achieved with an optimal tandem permutation (VP0–VP3–VP1), showing a protein yield close to 1:1:1. After SUMO removal at a low level of protease activity (5 units), the assembled FMDV virus-like particles (VLPs) could expose multiple epitopes and have a size similar to the naive FMDV. Immunization of pigs with the FMDV VLPs could induce FMDV-specific humoral and cellular immune responses effectively, in a dose-dependent manner. These data suggested that the stable FMDV VLPs with multiple epitope exposure were effective for the induction of an immune response in pigs, which laid a foundation for the further development of the FMDV subunit vaccine.

Background Candida is the common conditionally pathogenic fungus that infected human and animal clinically. C. tropicalis had been isolated from the skin and hair of healthy pigs, but with no report of fatal infection in gastrointestinal diseases. Case presentation In a pig farm in Henan Province of China, about 20 % of pregnant and postpartum sows suffered from severe gastrointestinal diseases, with a mortality rate higher than 60 % in the diseased animals. The sows had gastrointestinal symptoms such as blood in stool and vomiting. Necropsy revealed obvious gastric ulcers, gastrointestinal perforation, and intestinal hemorrhage in the gastrointestinal tract, but no lesions in other organs. The microbial species in gastric samples collected from gastric ulcer of the diseased sows then was initially identified as Candida by using routine systems of microscopic examination, culture characteristics on the medium Sabouraud dextrose agar medium. The fungus was further identified as C. tropicalis by species-specific PCR and sequencing. This study revealed an infection of C. tropicalis in sows through gastrointestinal mucosa could cause fatal digestive system disease and septicemia. Conclusions For the first time, a strain of C. tropicalis was isolated and identified from the gastric tissue of sows with severe gastrointestinal diseases. PCR and sequencing of ITS-rDNA combined with morphology and histopathological assay were reliable for the identification of Candida clinically.

为评估猪伪狂犬病病毒(Pseudorabies virus,PRV)灭活疫苗(HN1201-ΔgE株)免疫后对PRV流行毒株和经典毒株的保护效果,本研究对试验猪分别免疫PRV灭活疫苗(HN1201-ΔgE株)和PRV活疫苗(Bartha-K61),免疫后第0、7、10、14、17、21、24和28天采血测定PRV gB抗体,并分别使用PRV流行毒株HN1201株和经典毒株闽A株测定免疫后第0、7、14、21和28天血清的中和抗体水平,于免疫后第28天分别使用HN1201株和闽A株攻毒并观察,之后测定体温,测定攻毒后第7和14天PRV gE抗体,及攻毒后0～8 d的排毒情况.结果显示,HN1201-ΔgE免疫组较Bartha-K61免疫组gB抗体和中和抗体产生早,且抗体水平较高.两个免疫组试验猪在攻毒后虽然均无明显临床症状,且免疫组织化学检测(IHC)组织中的病毒抗原均为阴性,但HN1201-ΔgE免疫组试验猪脏器未见任何病理损伤,Bartha-K61免疫组试验猪部分脏器具有病理损伤.与未免疫对照组相比,2个免疫组试验猪在HN1201株和闽A株攻毒后,gE抗体转阳时间晚且排毒率低,HN1201-ΔgE免疫组gE抗体水平整体均低于Bartha-K61免疫组,攻毒后排毒检测中,Bartha-K61免疫组于2个毒株攻毒后第3～5天可检测到排毒,而HN1201-ΔgE免疫组全程未检测到排毒.研究结果表明,灭活疫苗(HN1201-ΔgE株)对PRV流行毒株和经典毒株均可提供完全保护.

2019年底新型冠状病毒(SARS-CoV-2)造成了对人类健康构成了巨大威胁的疫情,但其与猪冠状病毒基因组序列相似性低,在决定病毒宿主嗜性和毒力的纤突(spike,S)蛋白结构上具有较大差异,据此推测不能构成跨种传播.对蝙蝠携带冠状病毒多样性和分布区域的研究可有助于分析其对人类和养殖业的威胁,在公共卫生中具有积极意义.针对突发疫病,快速响应获得原始材料数据有助于疫病诊断、治疗和预防研究的开展.多种人冠状病毒S蛋白被证明是最适合开发的疫苗及治疗性抗体靶标,可在动物疫苗等产品的设计中借鉴.

Foot-and-mouth disease (FMD) is caused by FMD virus (FMDV) is a highly contagious disease of ruminants, which is primarily controlled by vaccination. The monitoring of antisera after vaccination is currently depending on liquid-phase blocking ELISA (LPBE). Recently, bacterium-original FMD virus-like particle (VLP) showed the potential as vaccine candidates. In this study, to minimize the risk of live virus involvement, the Escherichia coli original VLP of FMDV serotype O were used as the immunogen for monoclonal antibodies (Mabs) production and the capture antigen in the development of a solid-phase competition ELISA (SPCE). The samples with a percentage inhibition of >50% were considered positive in the SPCE assay. The concordance rate of the Mab-based SPCE compared with the LPBE for clinical serum samples test was 93.4%, and with a high agreement (kappa = 0.892) with LPBE in antibody duration monitoring. Results indicated that the VLP-based SPCE had high specificity and sensitivity, which provides an alternative method for postimmunization antibody evaluation of FMDV serotype O.

Foot-and-mouth disease (FMD), caused by FMD virus (FMDV), is a highly contagious epidemic disease, which is controlled primarily by prophylactic vaccination and serological monitoring after vaccination. Here, we have developed a solid-phase competition ELISA (SPCE) method based on virus-like particles (VLPs) of FMDV serotype A. The use of VLPs in the SPCE assay as a replacement for inactivated FMDV provides a high level of biosafety. The SPCE showed high concordance rates when compared with the virus neutralization test and liquid-phase blocking ELISA for testing clinical serum samples and successive serological monitoring (kappa = 0.925). Thus, this SPCE is an alternative method for post-immunization detection of antibodies against FMDV serotype A, with high specificity and sensitivity.

Rotavirus A (RVA) G9 genotype is recognized as an emerging genotype which is spreading worldwide, however, our knowledge on pathogenicity of this virus is limited. In this study, porcine RVA strain HN03 was successfully isolated on MA-104 cells, and the isolate was propagated continuously for 7 passages after a virus cloning at passage 3. The virus titers from 4 to 10 passages ranged from 10(7.1) to 10(8.1) TCID50/ml. The growth curve of HN03 strain in cell culture was determined, and the virus production dynamics was confirmed by immunoperoxidase monolayer assay (IPMA). Sequence and phylogenetic analyses based on full-length VP7 and partial VP4 genes indicated that HN03 strain belongs to genotype G9P[23]. In addition, the sixth passage of strain HN03 in cell culture was subjected to 3-day-old piglets. All infected piglets developed severe watery diarrhea within 24 hr post-inoculation (hpi), but recovered from disease after 72 hpi. RVA antigen could be detected by IHC in the cytoplasm of villous enterocytes as early as 2 hr after appearance of clinical symptoms and virus antigen load kept increasing in the next 30 hr. The dynamics of RVA HN03 strain proliferation on cells and in pigs extended our understanding of rotavirus pathogenicity.

Pseudorabies (PR) is an economically important viral disease of pigs which can infect numerous species of mammals including rodents. Commercial PR vaccines have been widely used worldwide to control and eradicate this disease. However, some PRV vaccines such as Bartha-K61 were occasionally reported to be lethal to mice. Since mice are commonly found in pig farms, the safety issue of PRV live vaccine across different species was never addressed. In this study, PRV vaccine strain Bartha-K61 was in vivo propagated in mice for five passages. The mortality of mice ranged from 80%-100% at each passage of PRV infection. The fifth passage of PRV was used to infect piglets to test its virulence on this species. The infected piglets clinically behaved normally and survived by the end of study (terminated at 10days post-infection). Histopathologically, there was infiltration of eosinophile granulocyte in tonsil and lung and no other changes were observed in other organs of infected pigs. Immunohistochemistry staining results showed that PRV antigen was only found in lung sample of one piglet. Therefore, the above results suggested there was no safety concern of Bartha-K61 PRV vaccine on pigs after the vaccine virus was passaged in mice for 5 times. The result of this study may suggest that mice may play a minimal role in the derivation of PRV vaccine-like field viruses that are believed to cause disease in young pigs.

As virulence-determining genes, RR1 and RR2 encode the small subunit and large subunit of viral ribonucleotide reductase (RR) in pseudorabies virus which have been extensively studied in mice. However, their role in pigs has not been adequately investigated. In this study, we deleted RR1 and RR2 genes based on a TK/gE/gI triple gene-deleted pseudorabies virus and tested its efficacy in pigs as a vaccine candidate. The rescued virus showed similar growth properties and plaque size in vitro as its parent strain. In an animal study, the virus could elicit humoral immune responses shown by generation of gB-specific antibodies and virus neutralizing antibodies. However, vaccination could not provide protection against virulent pseudorabies virus challenge since vaccinated pigs showed clinical pseudorabies-specific syndromes. The deficiency in protection may due to the generation of late and low levels of gB antibodies and virus neutralizing antibodies.

Background: Porcine cirovirus type 1 (PCV1) and type 2 (PCV2) are circulating in Chinese pig herds and the infected pigs develop antibodies to both viruses. Current commercial available ELISA kits cannot differentiate PCV2-specific antibodies from the mixtures of PCV1 and PCV2 antibodies in PCV1/2-infected or PCV2-vaccinated pigs. Therefore, the need for developing PCV2-specific ELISA methods is urgent to evaluate PCV2 antibody level in exclusion of PCV1 antibody interference after PCV2 vaccination.Results: Virus-like particles (VLPs) of PCV2 based on the recombinant Cap protein were expressed in Escherichia coli. A competing ELISA was established by using the VLPs as coating antigen and a PCV2-specific monoclonal antibody as the competing antibody. The competing ELISA was compared with the results obtained by using an immunoperoxidase monolayer assay on 160 serum samples. The sensitivity and specificity of this competing ELISA were determined as 96.5 and 96.0 %, at 2 standard deviation from the mean or 91.8 and 100 % at 3 standard deviations from the mean. Next, a serological survey of 1297 vaccinated serum samples collected from commercial pig herds in Beijing, Hunan and Henan provinces in China was conducted. The results showed that 85.9 % of sera having positive PCV2 antibodiesConclusions: The competing ELISA we developed in this study was both sensitive and specific to PCV2 and was suitable for large-scale PCV2 antibody monitoring in exclusion of PCV1 antibody interference after PCV2 vaccination.

We report the strategies leading to the large-production of soluble non-tag full-length porcine circovirus type 2 (PCV2) Cap protein in Escherichia coli. Under neutral pH condition, the purified recombinant Cap protein derived from E. coli expression self-assembles into homogenous round virus-like particle at the similar size of that of the intact PCV2 virus, which is further characterized by Cryo-EM single particle structure determined at 4.5Å. The engineered PCV2 rCap VLP was tested as a subunit vaccine for the protective efficacy against PCV2 challenge on 3-week old piglets. Similar to commercial available PCV2 vaccine, the Cap VLP-immunized piglets developed specific antibody-mediated response and were protected from the virulent SH PCV2 strain challenge. Hence, the production of E. coli based PCV2Cap-VLP could be applied as a cost-friendly and effective subunit vaccine to control PCV2 spreading in developing countries.

NADC30-like PRRSV has been recently reported and became endemic in vaccinated pig herds in China. The outbreaks of disease in vaccinated pigs indicated the inefficacy of commercial PRRSV vaccines. In this study, five commercial PRRSV vaccines that have been widely used in China were used to evaluate the efficacy to a NADC30-like PRRSV infection. The vaccinated pigs were challenged with HNjz15, a NADC30-like PRRSV at 28 days post vaccination. Compared to unvaccinated pigs, the vaccinated pigs clinically shortened the period of fever with less pig numbers of clinical manifestations and had improved body weight gain at the end of the study. However, the vaccinated pigs developed viremia with similar kinetics and suffered pathological lesions in lung and lymphoid tissues as the unvaccinated pigs. The virus load in tonsil, lung and lymph nodes detected by immunohistochemistry staining in vaccinated pigs was also similar to that in unvaccinated pigs which indicated the inability of vaccination to eradicate the virus from tissues of vaccinated pigs. Therefore, the above results suggested current commercial PRRSV vaccines could not provide complete protection to the NADC30-like PRRSV infection.

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The pathogenicity of HNjz15, an NADC30-like strain of porcine reproductive and respiratory syndrome virus (PRRSV), was investigated and compared to that of a highly pathogenic PRRSV JAX1 strain. Six-week-old pigs infected with each virus showed typical clinical symptoms, including high fever and respiratory disorders. Pigs infected with JXA1 had more-severe clinical manifestations than pigs infected with HNjz15. HNjz15 replicated in vivo with kinetics similar to those of JXA1 but induced a lower level of PRRSV-specific antibody at the beginning of virus infection. Histopathologically, JXA1 infection led to more-severe lung lesions and broader organ tropism than HNjz15 did. Different from what was observed with the previously reported NADC30-like PRRSV JL580 strain, all HNjz15-infected pigs survived until the end of the study. All of these results indicated that NADC30-like PRRSV HNjz15 is virulent to pigs but is less pathogenic than the JXA1 and JL580 PRRSV strains.