Lumpy Skin Disease Virus Infection Research Articles

Class IIa histone deacetylase (HDAC) inhibitor TMP269 suppresses lumpy skin disease virus replication by regulating host lysophosphatidic acid metabolism.

Lumpy skin disease virus (LSDV) infection poses a significant threat to global cattle farming. Currently, effective therapeutic agents are lacking. TMP269, a small molecule inhibitor of class IIa histone deacetylase inhibitor, plays a vital role in cancer therapy. In this study, we demonstrated that TMP269 treatment inhibits the early-stage replication of LSDV in a dose-dependent manner. RNA sequencing data revealed that metabolism-related signaling pathways were significantly enriched after LSDV infection. Furthermore, untargeted metabolomics analysis revealed that lysophosphatidic acid (LPA), a key metabolite of the glycerophospholipid pathway, was upregulated following LSDV infection and downregulated after TMP269 treatment. In addition, exogenous LPA promotes LSDV replication by activating the mitogen-activated protein kinase (MEK)/extracellular-signal-regulated kinase (ERK) signaling pathway and suppressing the host's innate immune response. Furthermore, treatment with the LPA receptor inhibitor Ki16425 suppressed LSDV replication and promoted the host's innate immune response. These findings suggest that LSDV infection can induce LPA expression and aid viral activation of the MEK/ERK signaling pathway and escape of the host's innate immune response, whereas TMP269 treatment can inhibit LPA production and limit its promotion of LSDV replication. These data identified the antiviral mechanism of TMP269 and a novel mechanism by which LSDV inhibits host innate immune responses, providing insights into the development of new preventive or therapeutic strategies targeting altered metabolic pathways.IMPORTANCELumpy skin disease virus (LSDV) poses a significant threat to global cattle farming. Owing to insufficient research on LSDV infection, pathogenesis, and immune escape mechanisms, prevention and control methods against LSDV infection are lacking. Here, we found that TMP269, a class IIa histone deacetylase inhibitor, significantly inhibited LSDV replication. We further demonstrated that TMP269 altered LSDV infection-induced host glycerophospholipid metabolism. In addition, TMP269 decreased the accumulation of lysophosphatidic acid (LPA), a key metabolite in glycerophospholipid metabolism, induced by LSDV infection, and exogenous LPA-promoted LSDV replication by activating the mitogen-activated protein kinase (MEK)/extracellular-signal-regulated kinase (ERK) signaling pathway and suppressing the host innate immune response. Our findings identified the antiviral mechanism of TMP269 and a novel mechanism by which LSDV manipulates host signaling pathways to promote its replication, offering insights into the development of novel antiviral agents against LSDV infection.

Molecular epidemiology and phylogenetic insights of lumpy skin disease in cattle from diverse agro-ecological regions of Punjab, Pakistan.

Lumpy skin disease (LSD) is an emerging, highly contagious transboundary disease of bovines caused by the Lumpy skin disease virus (LSDV), responsible for substantial economic losses to the dairy, meat, and leather industries in Pakistan as well as various countries around the world. Epidemiological information on LSD is scarce in Punjab, Pakistan. Therefore, a molecular epidemiological study was conducted in two agro-ecologically diverse districts (Bhakkar and Jhang) of Punjab, Pakistan. A total of 800 blood samples were randomly collected from the jugular vein of clinically suspected cattle with nodular lesions using a multistage cluster sampling technique. The sampling unit was indigenous, crossbred, and exotic breeds of cattle. Four hundred samples were collected from each district. Ten union councils (UC) were selected from each district, and two villages were selected from each union council. From each village, twenty cattle were selected for sample collection. The PCR-based overall prevalence of LSDV in clinically suspected cattle using the P32 gene was 36.25% (36.25%; 290/800). The multivariable logistic regression analysis indicated that animals who were not treated with acaricide (P = 0.014; OR = 1.459; C.I = 1.079-1.972), body condition score (emaciated animals; P = 0.019; OR = 1.573; CI = 1.076-2.301), and gender (female; (P = 0.016; OR = 1.435; CI = 1.072-1.969) were significantly at higher risk for LSDV infection in cattle. The phylogenetic insights revealed that our isolates were linked to Kenya, China, Russia, Egypt, India, Zimbabwe, Iraq, and Iran. It can be concluded that LSD is widely distributed in the study area, with evidence of genetic diversity. Further studies are required on genetic composition using variable genetic markers for effective control and eradication of LSDV in Pakistan.

Evidence of natural lumpy skin disease virus (LSDV) infection and genetic characterization of LSDV strains from water buffaloes (Bubalus bubalis) in India.

Lumpy skin disease (LSD) has emerged as a threat to cattle production in Asia, and India has been facing LSD epidemics since 2019. Although water buffalo (Bubalus bubalis) is susceptible to natural LSDV infection, there have been no confirmed reports of LSDV infection in water buffalo in India. In this study, we investigated suspected cases of LSD in water buffaloes from 12 Indian states and one union territory during 2020-2023. Buffaloes showed mild to moderate clinical disease with fever and nodular skin lesions, but most remained asymptomatic. Eighteen of 177 (10.18%) buffaloes in 12 districts in three states tested positive for LSDV by real-time PCR, while 22 of 57 (38.59%) from nine districts in six states tested positive for LSDV-neutralizing antibodies, demonstrating the prevalence of LSDV infection in buffaloes over a wider geographic area. Successful virus isolation and nucleotide sequencing confirmed natural LSDV infection in buffaloes. Phylogenetic analysis of complete GPCR, RPO30, and EEV gene sequences revealed the presence of wild-type strains of two divergent LSDV lineages (1.2.1 and 1.2.2) in buffaloes. The 1.2.2 strains were closely related to the dominant LSDV strain (subcluster 1.2.2, KSGP-like) circulating in India, while the 1.2.1 strains clustered with strains from the Middle East, Europe, and the Balkans, confirming that there have been multiple introductions of LSDV into India. The detection of viruses with identical sequences in buffaloes and local cattle implied that the LSDV strain found in buffaloes is probably of cattle origin. We also found evidence of cocirculation of LSDV 1.2.1 and 1.2.2 wild-type strains in the same area, highlighting the importance of LSDV surveillance and genetic analysis. This is the first confirmed report of natural LSDV infection in water buffaloes in India. Further investigations are needed to assess the impact of LSDV infection in buffalo production and the role of buffalo in LSD epidemiology.

Hematological, Biochemical and Cytokine Profiles in Cattle: Effects of Lumpy Skin Disease Virus Infection and Farm Size

Abstract Lumpy skin disease (LSD) is a contagious viral disease of cattle. This experiment aimed to study the influence of lumpy skin disease virus (LSDV) infection and farm size on hematological, biochemical parameters and cytokines in 55 beef cattle. The groups included LSDV–infected cattle from large farms (Group 1), LSDV–infected cattle from small farms (Group 2), uninfected cattle from large farms (Group 3), and uninfected cattle from small farms (Group 4). The hematological, biochemical values and cytokine profiles were measured. The results showed that red blood cells, hemoglobin, platelets, white blood cells, neutrophils, and lymphocytes were higher in infected cattle than in the uninfected cattle. The level of eosinophils of the cattle from the large farm was higher than that of the small farm. Gamma glutamyl transferase in the infected cattle was higher than in the uninfected cattle. Albumin of the cattle from the small farm was higher than that from the large farm. IFN–γ in infected cattle was higher than in uninfected cattle, while TNF–α in the infected cattle was lower than in the uninfected cattle. Our study indicated that LSDV infection altered hematological and biochemical parameters, including cytokine profiles, with farm size potentially influencing these alterations.

The expanding host range of lumpy skin disease virus in wild and domestic animals.

Clinical lumpy skin disease (LSD) predominantly affects cattle and, to lesser extent domestic water buffalos. Whilst earlier work focussed on the disease in Africa, the recent emergence of LSD virus (LSDV) as a major cause of disease in Asia has led to a widening range of susceptible hosts for the virus. This article lists the wild and domestic ungulates in which LSDV infection has been confirmed and considers the significance of the disease for these species in Asia.

Hematological and biochemical studies of lumpy skin disease virus infection in cattle

Hematological and biochemical studies of lumpy skin disease virus infection in cattle

Subclinical infection caused by a recombinant vaccine-like strain poses high risks of lumpy skin disease virus transmission.

Lumpy skin disease (LSD) is a transboundary viral infection, affecting cattle with characteristic manifestations involving multiple body systems. A distinctive characteristic of lumpy skin disease is the subclinical disease manifestation wherein animals have viremia and shed the virus through nasal and ocular discharges, while exhibiting no nodules but enlarged lymph nodes that are easily oversighted by inexperienced vets. Further research on the role of subclinically ill animals in the transmission of LSD virus (LSDV) can contribute to the development of more effective tools to control the disease worldwide. Thus, this study aims to determine the potential role of subclinical infection in virus transmission in a non-vector-borne manner. To achieve this, we inoculated animals with the recombinant vaccine-like strain (RVLS) Udmurtiya/2019 to cause clinical and subclinical LSDV infection. After the disease manifestation, we relocated the subclinically ill animals to a new clean facility followed by the introduction of another five animals to determine the role of RVLS-induced subclinical infection in the virus transmission via direct/indirect contact. After the introduction of the naïve animals to the relocated subclinically ill ones in a shared airspace, two introduced animals contracted the virus (clinically and subclinically), showing symptoms of fever, viremia, and seroconversion in one animal, while three other introduced animals remained healthy and PCR-negative until the end of the study. In general, the findings of this study suggest the importance of considering LSDV subclinical infection as a high-risk condition in disease management and outbreak investigations.

Development of membrane protein-based vaccine against lumpy skin disease virus (LSDV) using immunoinformatic tools.

Lumpy skin disease, an economically significant bovine illness, is now found in previously unheard-of geographic regions. Vaccination is one of the most important ways to stop its further spread. Therefore, in this study, we applied advanced immunoinformatics approaches to design and develop an effective lumpy skin disease virus (LSDV) vaccine. The membrane glycoprotein was selected for prediction of the different B- and T-cell epitopes by using the immune epitope database. The selected B- and T-cell epitopes were combined with the appropriate linkers and adjuvant resulted in a vaccine chimera construct. Bioinformatics tools were used to predict, refine and validate the 2D, 3D structures and for molecular docking with toll-like receptor 4 using different servers. The constructed vaccine candidate was further processed on the basis of antigenicity, allergenicity, solubility, different physiochemical properties and molecular docking scores. The in silico immune simulation induced significant response for immune cells. In silico cloning and codon optimization were performed to express the vaccine candidate in Escherichia coli. This study highlights a good signal for the design of a peptide-based LSDV vaccine. Thus, the present findings may indicate that the engineered multi-epitope vaccine is structurally stable and can induce a strong immune response, which should help in developing an effective vaccine towards controlling LSDV infection.

Identification of the murine osteoblastic cell MC3T3-E1 as a permissive cell line in response to lumpy skin disease virus

Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in China. Screening suitable cells for LSDV replication is vital for future research on pathogenic mechanisms and vaccine development. Previous comparative studies have identified that the rodent-derived BHK21 is a highly susceptible cell model to LSDV infection. Using western blot, indirect immune-fluorescence assay, flow cytometry, and transmission electron microscopy methods, this study is the first to identify the murine osteoblastic cell line MC3T3-E1 as a novel permissive cell model for LSDV infection. The establishment of MC3T3-E1 as a suitable infectious cell model enhances our understanding of the species range and cell types of the permissive cells and nonpermissive that support LSDV replication. It is helpful to accelerate future research on the pathogenesis, clinical application, and vaccine development of LSDV.

Emergence of lumpy skin disease virus (LSDV) infection in domestic Himalayan yaks (Bos grunniens) in Himachal Pradesh, India.

In this study, we investigated and confirmed natural lumpy skin disease virus (LSDV) infection in Himalayan yaks (Bos grunniens) in Himachal Pradesh, India, based on clinical manifestations and results of genome detection, antibody detection, virus isolation, and nucleotide sequencing. Subsequent phylogenetic analysis based on complete GPCR, RPO30, and EEV gene sequences revealed that the LSDV isolates from these yaks and local cattle belonged to LSDV subcluster 1.2.1 rather than the dominant subcluster 1.2.2, which is currently circulating in India, suggesting a separate recent introduction. This is the first report of natural LSDV infection in yaks in India, expanding the known host range of LSDV. Further investigations are needed to assess the impact of LSDV infection in yaks.

Emergence of Lumpy Skin Disease Virus Infection in Yaks, Cattle‐Yaks, and Cattle on the Qinghai–Xizang Plateau of China

Lumpy skin disease (LSD) is a viral disease caused by lumpy skin disease virus (LSDV), which mainly infects cattle and can cause huge economic losses. In May 2023, yaks, cattle‐yaks, and cattle in Tibet (Xizang), China, developed fever, skin nodules, and severe discharges and were suspected to be cases of LSD. Samples from these animals were analyzed using molecular biology and serological methods. The RPO30, P32, and GPCR genes were amplified by PCR and sequenced, and the whole genome of the virus was determined using viral metagenomics technology. Sequencing results showed that it was indeed an LSDV infection, and enzyme‐linked immunosorbent assay results confirmed the presence of LSDV antibodies. The whole genome phylogenetic tree shows that LSDV/CHINA/Tibet/2023 is different from the previous epidemic strains in China, but clusters with India 2022 strain. This is the first report of LSD in yaks, cattle‐yaks, and cattle on the highest altitude plateau in the world.

MiRNA profiling of primary lamb testicle cells infected with lumpy skin disease virus.

In this study, miRNA profiling of cells infected with lumpy skin disease virus (LSDV) was conducted for the first time. When compared to mock-infected cells, LSDV-infected primary lamb testicle (LT) cells showed dysregulation of 64, 85, and 85 miRNAs at 12 hours postinfection (hpi), 48 hpi, and 72 hpi, respectively. While some of these miRNAs were found to be dysregulated at a particular time point following LSDV infection, others were dysregulated at all three time points. Analysis of the differentially expressed miRNA-mRNA interaction networks, Gene Ontology analysis of the predicted targets, and KEGG analysis of highly enriched pathways revealed several cellular factors/pathways involved in protein/ion/enzyme binding, cell differentiation, movement of subcellular components, calcium reabsorption, aldosterone synthesis and secretion, and melanogenesis. Some selected upregulated (oar-mir-379-5p, oar-let-7d, Chr10-18769, Chr2_5162 and oar-miR-493-5p) and downregulated (ChrX-33741, Chr3_8257 and Chr26_32680) miRNAs were further confirmed by quantitative real-time PCR. These findings contribute to our understanding of virus replication, virus-host interactions, and disease pathogenesis, and the differentially expressed miRNAs and their cellular targets may serve as biomarkers as well as novel targets for therapeutic intervention against LSDV.

A Real-Time Recombinase Polymerase Amplification Assay for Specific Detection of Lumpy Skin Disease Virus.

Lumpy skin disease virus (LSDV) infection, accompanied by loss of hide quality, poor reproductive efficiency, consistent degenerative emaciation, and milk yield reduction of animals, causes severe economic implications in endemic zones. The heterologous attenuated goat pox (GTPV) vaccine (AV41 strain) was used in China to prevent LSDV infection. Only a few LSDV detection methods that distinguish LSDV from GTPV vaccine strains have been reported before. For simple, rapid, and specific detection of LSDV, the real-time recombinase polymerase amplification (RPA) method was established with the specific primers and probes designed according to the conserved regions of ORF132 gene sequences. The assay could be finished within 20 min at a constant temperature (39 °C). This method had a limit of detection (LOD) of 15 copies/μL for LSDV and no cross-reaction with the nucleic acids of goat pox virus, infectious bovine rhinotracheitis virus, Pasteurella multocida, and bovine healthy tissue. Furthermore, 43 clinical samples were detected by this method and the real-time PCR recommended by the World Organisation for Animal Health (WOAH), with a kappa value, was 0.94. These results demonstrated that the real-time RPA method for detecting LSDV developed in this study was characterized by high sensitivity and specificity, which has wide application value in the clinical diagnosis and detection of LSDV in China.

Lumpy Skin Disease Virus Infection Activates Autophagy and Endoplasmic Reticulum Stress-Related Cell Apoptosis in Primary Bovine Embryonic Fibroblast Cells.

Poxviruses have been associated with humans for centuries. From smallpox to mpox to lumpy skin disease virus (LSDV), members of the poxvirus family have continued to threaten the lives of humans and domestic animals. A complete understanding of poxvirus-mediated cellular processes will aid in the response to challenges from the viruses. In this study, we demonstrate that LSDV infection results in an abnormal ultrastructure of the endoplasmic reticulum (ER) lumen in primary bovine embryonic fibroblast (BEF) cells, and we further show that an ER imbalance occurs in LSDV-infected BEF cells. Additionally, we believe that ER stress-related apoptosis plays a role in the late apoptosis of BEF cells infected with LSDV, primarily through the activation of the CCAAT/enhancer binding protein homologous protein (CHOP)-Caspase-12 signal. In addition to cell apoptosis, a further investigation showed that LSDV could also activate autophagy in BEF cells, providing additional insight into the exact causes of LSDV-induced BEF cell death. Our findings suggest that LSDV-induced BEF cell apoptosis and autophagy may provide new avenues for laboratory diagnosis of lumpy skin disease progression and exploration of BEF cell processes.

Lumpy Skin Disease Virus Infection in Free-Ranging Indian Gazelles (Gazella bennettii), Rajasthan, India.

Near a zoo in Bikaner, India, 2 free-ranging Indian gazelles (Gazella bennettii) displayed nodular skin lesions. Molecular testing revealed lumpy skin disease virus (LSDV) infection. Subsequent genome analyses revealed LSDV wild-type strain of Middle Eastern lineage. Evidence of natural LSDV infection in wild gazelles in this area indicates a broadening host range.

Evaluation of haematological, serum biochemical and oxidative stress parameters in cattle naturally infected with lumpy skin disease virus.

Lumpy skin disease (LSD) is a vector-borne viral transboundary disease of cattle caused by the LSD virus (LSDV). Despite investigations on clinical and outbreak features of LSDV, information on disease pathogenesis and alternative changes in blood parameters are scarce. Keeping this in view, the current study was designed to determine haematological, serum biochemical, and oxidative stress parameters in naturally infected cattle with LSDV during the recent surge of outbreaks in Punjab, Pakistan. A total of 35 blood samples was collected from polymerase chain reaction-confirmed LSDV-infected cattle for assessment of all parameters. The haematological examination of blood samples showed a significant reduction (p < 0.05) in different variables of erythrogram and leucogram. On the other hand, differences between levels of various serum biochemical parameters with the significant increase in levels of alkaline phosphatase, serum alanine aminotransferase, aspartate aminotransferase, and blood urea nitrogen were observed in LSDV naturally infected cattle. Moreover, malondialdehyde levels for lipid peroxidation and nitrate concentration were markedly elevated whereas glutathione S-transferase fluorescent and serum superoxide dismutase enzymes showed a decrease in levels. The current study suggests that alternations in haematological and serum biochemical parameters following LSDV infection stimulate oxidative stress and such findings may be useful for early and rapid diagnosis and improvement in the treatment strategy of the disease.

The comparative study revealed that the hTERT-CSF cell line was the most susceptible cell to the Lumpy skin disease virus infection among eleven cells

Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in Asia, including China. Improving the propagation of LSDV is important for diagnostics and vaccine production. Our study identified and compared the LSDV susceptibility of eleven standard cells using western blot, indirect immune-fluorescence assay, quantitative PCR, and 50 % tissue culture infectious dose. Our finding revealed that the LSDV strain could infect five cell lines and show a cytopathic effect. Furthermore, the hTERT-CSF cell line had the highest level of virus in the five cell models, followed by BHK-21, MDBK, Vero, and hTERT-ST. Hence, hTERT-CSF could be used as a candidate cell line for basic and applied research, clinical application, and LSDV vaccine development, providing a vital reference in LSDV and other viruses.

Evidence of lumpy skin disease virus infection in camels

Countries in the Indian subcontinent are currently facing a deadly epidemic of lumpy skin disease (LSD). LSD is primarily a disease of cattle. Buffaloes may sometimes develop mild illness, however, other domestic animals are considered resistant to LSD. We confirmed the LSDV infection in camels as evidenced by skin nodules on the body surface of the affected camels, isolation of LSD virus (LSDV) and amplification of LSDV-specific gene segments from the skin nodules (PCR), nucleotide sequencing of the viral genome and, demonstration of anti-LSDV antibodies in serum. Phylogenetic analysis based on nucleotide sequencing of ORF011, ORF012 and ORF036 revealed that the virus (LSDV/Camel/India/2022/Bikaner) is related to the historical NI-2490/Kenya/KSGP-like field strains which are predominantly circulating in the Indian subcontinent. This is the first report wherein LSDV has been to infect camels.

Detection of Culex tritaeniorhynchus Giles and Novel Recombinant Strain of Lumpy Skin Disease Virus Causes High Mortality in Yaks.

Lumpy skin disease virus (LSDV) is capable of causing transboundary diseases characterized by fever, nodules on the skin, mucous membranes, and inner organs. The disease may cause emaciation with the enlargement of lymph nodes and sometimes death. It has had endemic importance in various parts of Asia in recent years, causing substantial economic losses to the cattle industry. The current study reported a suspected LSDV infection (based on signs and symptoms) from a mixed farm of yak and cattle in Sichuan Province, China. The clinical samples were found positive for LSDV using qPCR and ELISA, while LSDV DNA was detected in Culex tritaeniorhynchus Giles. The complete genome sequence of China/LSDV/SiC/2021 was determined by Next-generation sequencing. It was found that China/LSDV/SiC/2021 is highly homologous to the novel vaccine-related recombinant LSDV currently emerging in China and countries surrounding China. Phylogenetic tree analysis revealed that the novel vaccine-associated recombinant LSDV formed a unique dendrograms topology between field and vaccine-associated strains. China/LSDV/SiC/2021 was found to be a novel recombinant strain, with at least 18 recombination events via field viruses identified in the genome sequence. These results suggest that recombinant LSDV can cause high mortality in yaks, and its transmission might be due to the Culex tritaeniorhynchus Giles, which acts as a mechanical vector.

Seroepidemiological Survey of Lumpy Skin Disease among Cattle in El Gezira State (Central Sudan)

Lumpy skin disease (LSD) is an infectious viral disease that affects cattle, buffalos and selected wild animals. A cross-sectional study was conducted in El Gezira State, Central Sudan to determine the seroprevalence and risk factors of LSD in cattle in seven localities. A total of 197 serum samples were collected randomly from apparently healthy cattle during the period between April – May, 2021 and tested serologically with a commercial ELISA kit. The overall true seroprevalence of lumpy skin disease virus (LSDV) was 24.6% (50/198). Univariate analysis showed that there was a significant difference (p &lt; 0.05) in seropositivity to LSD between age groups, sex as well as breed. Multivariate analysis revealed that only two independent risk factors (age and breed) were statistically significant (p &lt; 0.05). It is apparent from the present study that LSDV infection is quite prevalent among cattle in El Gezira State, Central Sudan. To the best of the author’s knowledge, this is the first seroepidemiological study of LSDV infection in Central Sudan. The authors recommend that the concerned authorities expend more efforts to control this economically important disease in Sudan.