Porcine circovirus type 2d (PCV2d) is one of the predominant genotypes associated with porcine circovirus-associated diseases, which highlights the need for effective vaccination strategies. This study assessed the expression profile of the major histocompatibility complex class II transactivator (CIITA) gene in pigs immunized with recombinant PCV2d virus-like particles (VLPs). Recombinant PCV2d capsid protein was expressed in insect cells and self-assembled into VLPs, which were further used to immunize piglets. Peripheral blood mononuclear cells were analyzed for CIITA expression by qRT-PCR and interferon-gamma (IFN γ) production by ELISA. Following PCV2d VLP immunization, CIITA expression increased over time, peaked on day 21, and was associated with elevated IFN-γ secretion. A significant positive correlation between CIITA expression and IFN γ levels was detected in pooled analyses. These findings demonstrate that PCV2d VLPs endorse Th1-biased immune responses and further enhance the CIITA-mediated antigen presentation, supporting their potential as an effective vaccine platform against PCV2d.
Porcine circovirus type 2d (PCV2d) is the predominant genotype etiological agent of porcine circovirus-associated disease (PCVAD), causes substantial economic losses to the swine industry worldwide, including in India. Since currently available vaccines are mainly based on the PCV2a genotype, which is unable to offer cross protective immunity against emerging PCV2d strains, development of PCV2d specific vaccine is necessary. In this study, an indigenous virus-like particle (VLP)-based vaccine candidate was developed using the capsid coding gene (orf2) of an Indian PCV2d isolate expressed in a baculovirus-insect cell system. Recombinant capsid protein expression and self-assembly into VLPs were confirmed by Western blotting and transmission electron microscopy. The purified VLPs showed favourable thermostability as well as pH stability and elicited strong humoral and cellular immune responses in in vivo porcine model. A delayed booster immunization strategy resulted in prolonged antibody persistence. In further, in vivo challenge study with virulent homologous PCV2d virus vaccinated animals were protected against PCV2 clinical signs. Further, absence of detectable viral DNA in swab and tissue samples, and markedly alleviated histopathological lesions compared with unvaccinated controls animals. Overall, these findings suggest that the indigenous PCV2d VLP vaccine candidate is highly immunogenic and provides protective immunity against homologous PCV2d challenge, highlighting its potential indigenous vaccine candidate for controlling PCVAD against PCV2d.
African Swine Fever (ASF) causes severe economic losses in the global pig industry, characterized by high mortality rates and lack of an effective vaccine. Interestingly, India's indigenous Doom pig breed has displayed tolerance to ASF, remaining seropositive without detectable viremia. This study investigated the genetic basis of ASFV tolerance, focusing on the STING1 gene, a key component of the cGAS-STING antiviral signaling pathway. In-silico docking identified a potential ASFV protein binding site within exon 5 of STING1. A non-synonymous SNP (CGG/TGG; R→W) at position 148 in this region was selected for genotyping. We genotyped 119 pigs, representing tolerant (Doom), susceptible, and ASFV-infected groups, using allele-specific PCR and Sanger sequencing. Doom pigs and most other breeds, including ASFV-infected samples, displayed a conserved GG genotype. Susceptible Manipuri Black and Ghoongroo pigs, however, displayed both AA and GG genotypes, yet still succumbed to the disease. A significant downregulation (0.436-fold) of STING1 was observed in ASFV-infected spleen tissue, indicating active immune evasion by the virus. The lack of a unique allele in Doom pigs compared to susceptible breeds indicates that ASFV tolerance is unlikely to be associated with this STING1 SNP. Therefore, genome-wide studies are recommended to identify markers truly associated with ASFV tolerance in this resilient breed.
Anti-Mullerian hormone (AMH) is a biomarker of reproductive lifespan in cows. In our previous study, we developed an in-house ELISA for bovine AMH. Afterwards, we targeted to validate this ELISA in field condition and to compare the efficacy of this ELISA with other commercial assays. Also, we aimed to standardize the age-specific AMH level in the Bos indicus cows using both assays. For this, blood samples were collected from cows of 0–10 yrs of age (Sahiwal and Tharparkar breed). The minimum AMH level was found at 0–6 months of age (199.07 ± 44.89 and 180.87 ± 32.76 pg/mL for each breeds. Afterwards, it increased significantly (p < 0.05) and reached a peak value between 1.5–2.0 and 2.0–2.5 years in Sahiwal and Tharparkar (1497.71 ± 77.18 1725.14 ± 114.81 pg/mL). Afterwards, it starts decreasing with age. Both the assays showed high correlation. The age-specific profiling of AMH was done for the first time in indigenous cows and the in-house ELISA was found as efficient as the commercial assay.
Pigs serve as critical reservoirs and amplifiers for numerous zoonotic viral diseases, presenting substantial public health challenges in India. This study highlights the epidemiology and emerging trends of key zoonotic viruses associated with pigs, emphasizing their role in endemic and emerging disease dynamics. Japanese encephalitis virus (JEV) persists as a major concern, with pigs acting as amplifying host, while hepatitis E virus (HEV) remains a prominent cause of viral hepatitis, transmitted via contaminated water and pork products. Emerging high-fatality viral zoonoses caused by Nipah virus (NiV) and recurrent threats from swine influenza virus (SIV) demonstrate that the zoonotic landscape is evolving. Furthermore, zoonotic viruses like rotavirus, pseudorabies (ADV or SuHV-1), porcine astrovirus (PAstV), and Torque teno sus virus (TTSuV) reflect the expanding diversity of pig-associated pathogens in India. Emerging evidence also implicates viruses such as Chandipura virus (CHPV) in localized outbreaks, indicating broader zoonotic potential. Novel risks such as swine acute diarrhea syndrome coronavirus (SADS-CoV) and SARS-CoV-2 emphasize the role of pigs as potential intermediaries for pandemic-prone viruses. This comprehensive study evaluates the prevalence, outbreak dynamics, and public health implications of zoonotic viral diseases of pigs in India, providing valuable direction for developing effective control measures.
This article considers the sparse solutions of the tensor complementarity problem (TCP). For a TCP with Z-tensor we establish a connection between the least element of the feasible set of TCP and the sparse solution of the TCP. We propose a p norm regularized minimization problem and show that it can approximate sparse solution applying the regularization of parameter.
This research aimed to characterize the mitochondrial genome of the Ghoongroo (GH) pig, a notable breed in India, along with its crossbred varieties, to elucidate their matrilineal components, evolutionary history, and implications for conservation. Seven pigs (5 GH, 2 crossbred, namely Rani and Asha) were sequenced for complete mitochondrial genome, while 24 pigs (11 GH, 6 Rani, and 7 Asha) were sequenced for the complete D-loop of the mitochondrial genome. The genome size of these pigs was determined to be 16,690 bp. Analysis of the mitochondrial sequences and phylogenetics uncovered two distinct matrilineal components within the GH population, a phenomenon also observed in its crossbred counterparts, Rani and Asha. Phylogenetic analysis demonstrated a clear clustering of GH sequences into two clades, indicating the presence of two independent maternal lineages. The phylogenetic study using complete mitogenome also indicated that GH pigs were originated locally from Indian wild boar independently from Asian and European pig population. Haplotype analysis from complete D-loop sequences revealed 10 different haplotypes, with some sequences shared among GH, Rani, and Asha, while others differed due to varying matrilineal origins. The haplotype analysis using complete mitogenome sequences revealed 16 different haplotypes with some shared sequences among GH. Furthermore, examination of tRNA genes and nucleotide composition of different genes namely rRNAs, COX1, COX2, ATP6, ND4, ND5, ND6, Cytb offered insights into genetic diversity within these pigs. The findings suggest that geographical isolation and historical events likely contributed to the emergence of distinct maternal lineages within the GH breed. This study underscores the significance of mitochondrial DNA analysis in uncovering hidden genetic diversity within seemingly uniform populations. The molecular insights gained into the genetic makeup of GH pigs could aid in designing effective breeding programs for conservation efforts and highlight its significance in understanding the broader context of pig domestication in India.
African swine fever (ASF) is a devastating viral disease in pigs, caused by an enveloped DNA virus (ASFV), with 100
In this article, we introduce column adequate tensor in the context of tensor complementarity problem and consider some important properties. The tensor complementarity problem is a class of nonlinear complementarity problem with the involved function being defined by a tensor. We establish the inheritance property and invariance property of column adequate tensors. We study TCP (q,𝒜) in the frame of LCP (q̅,A̅) . We show that TCP (q,𝒜) has ω -unique solution under some assumptions.
Since the first described outbreak of African Swine Fever (ASF), which is caused by African swine fever virus (ASFV), in Kenya, over a century ago, no approved commercial vaccine is widely available to prevent the disease. Research has focused on two main aspects of the disease. Firstly, targeting the virus virulence factors by identifying proteins responsible for host immune evasion that can be targeted to develop vaccine candidates. Novel strategies, including mRNA-based vaccines and synthetic ASFV genomes with reverse genetics, enable safe, immunogenic, and rapid development of live-attenuated and subunit vaccine candidates. Secondly, by identifying host determinants that confer resistance to ASF. ASFV infects both domestic pigs and wild boars irrespective of age and breed, causing nearly 100 % mortality. Warthogs and bushpigs, natural hosts of ASFV, show no clinical signs despite developing viremia, likely due to distinct innate and adaptive immune responses, epigenetic modifications, and environmental factors. Genetic and epigenetic investigations of this tolerance, focusing on type I interferon (IFN) induction and NF-κB pathways, which are often targeted by viral immune evasion proteins. Understanding these interactions and genetic variations between tolerant/resistant and susceptible animals may guide vaccine development and the creation of tolerant/resistant breeds. This review covers the ASFV genome, transmission, pathogenesis, virus's immune evasion strategies, and the host immunological response against ASFV.
African Swine Fever (ASF) is a highly contagious transboundary viral disease affecting domestic pigs worldwide, often resulting in nearly 100 % mortality due to the lack of effective vaccines. However, wild species such as warthogs (Phacochoerus sp.) and bush pigs (Potamochoerus sp.) do not exhibit clinical symptoms of the disease, previous studies showed that amino acid substitutions in a proto-oncogene, RelA, a subunit of NF-κB found in warthogs might be responsible for their resistance. Expanding this study over an Indian breed named Doom which was considered tolerant due to lack of much information regarding their ASFV-positive cases and identifying the genetic basis of tolerance might help in creating a tolerant breed, thereby controlling the spread of the disease. So, this study was initiated to investigate the polymorphic signature in RelA gene of Doom breed similar to warthogs. Initially molecular docking studies identified a potential interaction between the N-terminal sub-domain of Rel homology domain of porcine RELA and the African Swine Fever Viral (ASFV) protein A238L, a hypothetical viral protein of unknown function. This amino acid interaction corresponds to a nucleotide sequence of exon 10 of porcine RelA gene. Further, ARMS-PCR (Amplification Refractory Mutation System-PCR) and PCR-RFLP (Restriction Fragment Length Polymorphism) were optimized to target a single nucleotide polymorphism (SNP, CCT/GCT: Pro→Ala) identified in the exon 10 region through Ensemble genome browser. PCR results showed the presence of homozygous CC genotype, coding for proline, in all pig breeds tested, including Doom. Sanger sequencing confirmed the CC genotype across all breeds, indicating that amino acid substitutions in this RELA domain were not present in Doom and therefore may not be responsible for ASF tolerance. Since disease tolerance is a polygenic trait relying on one gene might not reveal the exact cause for their tolerance against ASFV. Future research can be targeted on other set of genes playing important role in innate immune pathways or Genome Wide Association Study can be preferred to identify and associate SNPs with Doom's tolerance against ASFV.
Classical Swine Fever (CSF) is a highly contagious viral disease that causes substantial economic losses in the swine industry. Timely detection of CSFV infection is essential for effective disease control; however, current antibody-based diagnostic methods are limited due to the delayed host immune response. In this study, we developed a novel peptide-based lateral flow assay (LFA) targeting the E2 antigen of the Classical Swine Fever Virus (CSFV) to enable rapid and field-deployable detection. A synthetic peptide derived from the immunodominant region of the CSFV E2 protein was identified through bioinformatics analysis, synthesized, and conjugated to Keyhole Limpet Hemocyanin (KLH) to enhance its immunogenicity. Polyclonal antibodies were generated in rabbits, purified, and validated via ELISA to confirm specificity to CSFV, showing no cross-reactivity with other swine viruses such as ASFV, PRRSV, PCV2, and PPV. Silver nanoparticles (AgNPs) were biosynthesized using Mangifera indica leaf extract and characterized by UV-vis spectroscopy, FTIR, and dynamic light scattering (DLS). These nanoparticles were then conjugated to the purified anti-E2 antibodies and incorporated into LFA strips. The resulting assay exhibited high sensitivity, capable of detecting CSFV antigen concentrations as low as 20 ng/ml, with excellent specificity and no cross-reactivity. This peptide-based LFA provides a rapid, reliable, and cost-effective diagnostic tool for on-site CSF surveillance and outbreak management.
Escherichia coli and Staphylococcus aureus are the most important food borne pathogen transmitting from animal meat and meat products. Therefore, it is vital to design an accurate and specific diagnostic tool for identifying those food-borne pathogens in animal meat and meat products. In the current study, E. coli, methicillin-resistant and sensitive S. aureus (MRSA and MSSA) were simultaneously detected using a developed triplex PCR-based technique. To obtain an optimal reaction parameter, the multiplex assay was optimised by changing just one parameter while holding the others constant. Specificity of the assay was assessed using several porcine bacterial template DNA. The plasmid DNA was used to test the multiplex PCR assay’s sensitivity and interference in spiked pork samples. E. coli, MRSA, and MSSA each have PCR amplified products with sizes of 335, 533, and 209 bp, respectively. The assay detects a minimum microbial load of 102 CFU/μl for all the three pathogens and can identify bacterial DNA as low as 10−2 ng/µl. The assay was validated employing 210 pork samples obtained from retail meat shops and slaughter houses, with MRSA, E. coli, and MSSA with the occurrence rate of 1.9
Mitochondrial DNA (mtDNA) serves as a valuable molecular marker for constructing matrilineal genealogies and tracing the evolutionary history of animals. This study aimed to characterize the complete mitochondrial genome of the Indian wild pig (IWB) (Sus scrofa cristatus) and identify IWB-specific DNA sequences that could be used as genomic signatures to differentiate IWB from domestic Indian pigs (IDP) in forensic cases. For the purpose, three wild IWB from a rescue centre were used for the characterization of the mitochondrial genome of the IWB. The mitochondrial genome was sequenced by the primer walking technique using 30 overlapping primers. The mitochondrial genome of the IWB was found to be 16,689 bp long containing 37 genes coding for 2 rRNAs, 22 tRNAs, 13 protein coding genes, and 1 D-loop region similar to the mitogenome of other pigs. Sequence analysis of the D-loop of IWB with other IDP indicated some signature sequence for IWB like duplication and transition event from 1090th to 1099th position, deletion of a 10 bp sequence at the 755th position, insertion of (CA) at the 137th position, and substitution of AT to GA at the 638th position. These variations specially the duplication along with transition event causes creation of unique signature sequence (-ACACAAACCT-) in the IWB that could serve as signature sequences for the IWB and be used as markers for differentiation of IWB from IDP breeds in academic as well as forensic or vetero-legal cases. Overall, a total of 36 polymorphic positions were identified in the IWB, with 29 sites being unique to the IWB only and seven being common to the Doom and HDK75 pig breeds. None of the common polymorphic sites were identified in prevailing domestic pig populations. Phylogenetic analysis of the mitochondrial genome revealed the distinct separation of the IWB from IDP. The results of genetic distance evaluation showed that the Doom pig breed was the closest to the IWB. This study provides valuable insights into the mitogenome characterisation, signature sequence and genetic distance analysis of the IWB and establishes a foundation for future studies on the conservation of this protected species.
A culture-free herbal aided assay designed for rapid detection of Extended spectrum β-lactamase producing bacterial isolates in piggery farm and slaughter house waste samples. The assay was validated with 309 piggery farm and slaughterhouse waste samples. The relative accuracy and relative sensitivity of the developed assay was found to be 85.1
Since 2020, African swine fever (ASF) has affected all pig breeds in Northeast India except Doom pigs, a unique indigenous breed from Assam and the closest relatives of Indian wild pigs. ASF outbreaks result in significant economic losses for pig farmers in the region. Based on sequencing and phylogenetic analysis of the B646L (p72) gene, it has been determined that ASFV genotype II is responsible for outbreaks in this region. Recent studies have shown that MYD88, LDHB, and IFIT1, which are important genes of the immune system, are involved in the pathogenesis of ASFV. The differential expression patterns of these genes in surviving ASFV-infected and healthy Doom breed pigs were compared to healthy controls at different stages of infection. The ability of Doom pigs to withstand common pig diseases, along with their genetic resemblance to wild pigs, make them ideal candidates for studying tolerance to ASFV infection. In the present study, we investigated the natural resistance to ASF in Doom pigs from an endemic area in Northeast India. The results of this study provide important molecular insights into the regulation of ASFV tolerance genes.
In this paper, we introduce set-valued tensor complementarity problem where the elements of the involved tensors are defined based on a set-valued mapping. We study several properties of the solution set under the framework of set-valued mapping. We provide the necessary and sufficient conditions for the zero solution of a set-valued tensor complementarity problem. We introduce limit $R_0$-property for the set of tensors and establish a connection between limit $R_0$-property and the level boundedness of the merit function of the corresponding set-valued tensor complementarity problem.
Viral infection disrupts the normal regulation of the host gene’s expression. In order to normalise the expression of dysregulated host genes upon virus infection, analysis of stable reference housekeeping genes using quantitative real-time-PCR (qRT-PCR) is necessary. In the present study, healthy and African swine fever virus (ASFV) infected porcine tissues were assessed for the expression stability of five widely used housekeeping genes (HPRT1, B2M, 18 S rRNA, PGK1 and H3F3A) as reference genes using standard algorithm. Total RNA from each tissue sample (lymph node, spleen, kidney, heart and liver) from healthy and ASFV-infected pigs was extracted and subsequently cDNA was synthesized, and subjected to qRT-PCR. Stability analysis of reference genes expression was performed using the Comparative delta CT, geNorm, BestKeeper and NormFinder algorithm available at RefFinder for the different groups. Direct Cycle threshold (CT) values of samples were used as an input for the web-based tool RefFinder. HPRT1 in spleen, 18 S rRNA in liver and kidney and H3F3A in heart and lymph nodes were found to be stable in the individual healthy tissue group (group A). The majority of the ASFV-infected organs (liver, kidney, heart, lymph node) exhibited H3F3A as stable reference gene with the exception of the ASFV-infected spleen, where HPRT1 was found to be the stable gene (group B). HPRT1 was found to be stable in all combinations of all CT values of both healthy and ASFV-infected porcine tissues (group C). Of five different reference genes investigated for their stability in qPCR analysis, the present study revealed that the 18 S rRNA, H3F3A and HPRT1 genes were optimal reference genes in healthy and ASFV-infected different porcine tissue samples. The study revealed the stable reference genes found in healthy as well as ASF-infected pigs and these reference genes identified through this study will form the baseline data which will be very useful in future investigations on gene expression in ASFV-infected pigs.