Lumpy Skin Disease Virus Infection Research Articles

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.

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.

LSDV126 gene based molecular assays for specific detection and characterization of emerging Lumpy Skin Disease virus

Lumpy skin disease (LSD) is a highly infectious and economically important viral disease, which is currently emerging in the Indian subcontinent. LSD is caused by Lumpy Skin Disease Virus (LSDV) under the genus Capripoxvirus and the family Poxviridae. Since its first incursion in India in the year 2019, the virus is rapidly disseminating through different means like direct contact, fomites and mainly by blood-feeding insects. As the disease has never been reported from India or neighbouring countries, there is a lack of planning and preparatory measures in terms of diagnostics and vaccines to control the disease. In the absence of any homologous vaccine, a live attenuated heterologous goat pox vaccine (Uttarkashi strain) is now being widely used in the country for the prevention of LSDV infection. Use of live attenuated goat pox virus vaccine necessitates the availability of an assay which could specifically detect and differentiate LSDV from goat pox virus. In this study, nucleotide sequences of LSDV126 gene encoding extracellular enveloped virus protein of circulating LSDV and goat pox virus were determined and analyzed. Deletion of 27 nt tandem repeats was observed in LSDV in comparison to goat pox and LSDV vaccine viruses. The deletion region was targeted for designing primers specific to LSDV, but not goat pox virus. A novel isothermal polymerase spiral reaction (PSR) was optimized as pen side diagnostic for prompt and sensitive detection of genomic DNA of LSDV. The assay was found to be highly sensitive and specific when compared to the real-time PCR. The assay was found to be specifically detecting only LSDV but not the goat pox virus. The limit of detection was identified as 9 × 10−6 ng of positive DNA. The assay will provide a point of care tool that will be a boon for the successful control of LSD in India.

Assessing machine learning techniques in forecasting lumpy skin disease occurrence based on meteorological and geospatial features.

Lumpy skin disease virus (LSDV) causes an infectious disease in cattle. Due to its direct relationship with the survival of arthropod vectors, geospatial and climatic features play a vital role in the epidemiology of the disease. The objective of this study was to assess the ability of some machine learning algorithms to forecast the occurrence of LSDV infection based on meteorological and geological attributes. Initially, ExtraTreesClassifier algorithm was used to select the important predictive features in forecasting the disease occurrence in unseen (test) data among meteorological, animal population density, dominant land cover, and elevation attributes. Some machine learning techniques revealed high accuracy in predicting the LSDV occurrence in test data (up to 97%). In terms of area under curve (AUC) and F1 performance metric scores, the artificial neural network (ANN) algorithm outperformed other machine learning methods in predicting the occurrence of LSDV infection in unseen data with the corresponding values of 0.97 and 0.94, respectively. Using this algorithm, the model consisted of all predictive features and the one which only included meteorological attributes as important features showed similar predictive performance. According to the findings of this research, ANN can be used to forecast the occurrence of LSDV infection with high precision using geospatial and meteorological parameters. Applying the forecasting power of these methods could be a great help in conducting screening and awareness programs, as well as taking preventive measures like vaccination in areas where the occurrence of LSDV infection is a high risk.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11250-022-03073-2.

Diagnosis of naturally occurring lumpy skin disease virus infection in cattle using virological, molecular, and immunohistopathological assays.

Background and Aim:Lumpy skin disease (LSD) is a contagious viral disease that has great economic losses among Egyptian breeding flocks. The present study was designed to compare the results of different diagnostic approaches used for the diagnosis of LSD virus (LSDV).Materials and Methods:A total of 73 skin nodule samples were collected from suspected infected cattle with LSDV from some Egyptian governorates during 2019 and 2020. Trials for virus isolation (VI) and identification on embryonated chicken eggs (ECEs) were conducted. Molecular detection, histopathological, and immunohistochemical examination were also conducted.Results:The virus was isolated into ECEs, and 58 samples of 73 were positive and gave a characteristic pock lesion on the chorioallantoic membrane. Twenty-two representative nodular skin specimens of the 58 positive samples were selected to be used for molecular, histopathological, and immunohistochemistry (IHC) diagnosis. Conventional polymerase chain reaction succeeded in detecting LSDV DNA in all tested 22 skin nodule samples. Histological examination of skins of different cases revealed various alterations depending on the stage of infection. IHC was used as a confirmatory test for detecting LSDV antigen in the tissues of the skin nodules of infected cattle using specific anti-LSDV antibodies. Lumpy skin viral antigen was detected within the cytoplasm of the epidermal basal cells layer and prickle cell and within the cytoplasm of the hair follicles’ epithelial outer and inner roots.Conclusion:This study confirmed the prevalence of LSDV infection in different Egyptian governorates during 2019 and 2020. In addition, histopathology and IHC could be potential methods to confirm Lumpy skin disease infection besidesVI and molecular detection.

Minimum Infective Dose of a Lumpy Skin Disease Virus Field Strain from North Macedonia

Infection with Lumpy Skin Disease virus (LSDV), as well as infections with other Capripox virus species, are described as the most severe pox diseases of production animals and are therefore listed as notifiable diseases under the guidelines of the World Organization for Animal Health (OIE). To our knowledge there is only a single study examining dose dependency, clinical course, viremia, virus shedding, as well as serological response following experimental LSDV “Neethling” inoculation. Here, we inoculated cattle with four different doses of LSDV strain “Macedonia2016”, a recently characterized virulent LSDV field strain, and examined clinical symptoms, viremia, viral shedding, and seroconversion. Interestingly, around 400 cell culture infectious dose50 (CCID50) of LSDV-“Macedonia2016” were sufficient to induce generalized Lumpy Skin Disease (LSD) in two out of six cattle but with a different incubation time, whereas the other animals of this group showed only a mild course of LSD. However, differences in incubation time, viral loads, serology, and in the clinical scoring could not be observed in the other three groups. In summary, we concluded that experimental LSDV infection of cattle with an infectious virus titer of 105 to 106 CCID50/mL of “Macedonia2016” provides a robust and sufficient challenge model for future studies.

Lumpy Skin Disease Virus Infection: An Emerging Threat to Cattle Health in Bangladesh

Lumpyskin disease (LSD) is a highly infectious and economically important transboundary disease of cattle caused by LSD virus (LSDV) LSD is characterized by fever and the appearance of nodular lesions in the skin covering all parts of the body LSD is currently endemic in many African countries and causing significant economic losses LSD has recently spread out of Africa into the Middle East region, and still continuing to spread other territories of the world In September 15, 2019 the first occurrence of LSD outbreak in Bangladesh has been reported to the OIE (World Organization for Animal Health) LSD is a vector-borne disease, and climatic conditions of Bangladesh are favorable for the propagation of vectors, which should influence the rapid transmission of LSD in the cattle population of Bangladesh Therefore, LSD may appear as a big challenge to cattle health in Bangladesh in the near future Consequently, it is very important to take strategic plans and immediate control measures for LSD outbreak control and prevention in Bangladesh In this review article, we will focus on epidemiology and modes of LSDV transmission, and its effective control and preventive measures to reduce or eradicate the LSDV infection from a recently infected country like Bangladesh In addition, we will highlight on impacts of LSD outbreaks in cattle industry of Bangladesh, and necessity of farmers’ awareness building and telemedicine service during the Coronavirus disease 2019 (COVID-19) outbreaks in controlling LSD

Molecular characterization of the 2018 outbreak of lumpy skin disease in cattle in Upper Egypt.

Background and Aim:Lumpy skin disease (LSD), an infectious disease of cattle, is characterized by raised nodules on the skin. Although the morbidity rate of LSD is low, it has a considerable fatality rate. Despite the annual mass vaccination of livestock with sheep pox vaccine (Veterinary Serum and Vaccine Research Institute, Egypt) enforced by Egyptian authorities, the LSD virus (LSDV) continues to circulate almost every summer. The present study aimed to discover the cause of cows naturally infected with LSDV circulating in Upper Egypt during the summer of 2018 using polymerase chain reaction (PCR) assay and to analyze their phylogenetics against reference genome sequences.Materials and Methods:We cultured LSDV in specific pathogen-free embryonated chicken eggs (SPF-ECE) and used conventional PCR to identify fusion and P32 genes, previously deposited in GenBank (MN694826, MN694827, and MN954664). Sequencing and phylogenetic analyses were performed on these two highly conserved viral genes.Results:LSDV infection of SPF-ECE resulted in characteristic white pock lesions. PCR products were identified on 1.5% agarose gel after electrophoresis at the expected positions for the fusion and P32 genes at 472 and 587 bp, respectively.Conclusion:The present study revealed that the two viral genes were identified from the Beni Suef and Sohag Governorates in all clinical cases and confirmed the circulation of LSDV in this outbreak. After sequencing, these genes were identical to those of the LSDV that had been identified and recorded in GenBank for the past 3 years.

Lumpy skin disease virus: transmission to dipteran vectors using animal and ex-vivo models

Lumpy skin disease (LSD) is a tropical neglected viral disease of cattle, characterised by numerous cutaneous lesions disseminated throughout the body. Historically endemic to the African continent, it has become a threat to Europe following the outbreaks of LSD in the Middle East and Eastern Europe. LSD virus (LSDV) is a Capripoxvirus transmitted by insect vectors. Experimental and epidemiological studies have indicated a role for the stable fly (Stomoxys calcintrans) and the mosquito Aedes aegypti. Nevertheless the relative importance of these vector species and others is unclear. A study was designed to explore the risk of transmission of LSDV from cattle to different vector species including Aedes aegypti, Culex quinquefasciatus, Stomoxys calcitrans and Culicoides nubeculosus. Cattle was challenged with LSDV to produce a bovine experimental model used as a natural source of LSDV to the potential vectors. Cattle samples were taken to quantify LSDV in different tissues and characterise the disease. All insect species were allowed to feed on LSDV-challenged cattle at regular intervals and incubated for up to eight days. This data was then used to model the dynamics of LSDV infection and transmission. All four species were able to acquire and maintain LSDV for up to eight days post feeding, and the risk of transmission from bovine donor to insect was dependent on the severity of the disease. A model was then generated using ex-vivo skin lesions and infectious blood that will allow further studies of the role of these vectors.