Poxvirus Infections in Dairy Farms and Transhumance Cattle Herds in Nigeria.

BackgroundPoxviruses within the Capripoxvirus, Orthopoxvirus, and Parapoxvirus genera can infect livestock, with the two former having zoonotic importance. In addition, they induce similar clinical symptoms in common host species, creating a challenge for diagnosis. Although endemic in the country, poxvirus infections of small ruminants and cattle have received little attention in Botswana, with no prior use of molecular tools to diagnose and characterize the pathogens.MethodsA high-resolution melting (HRM) assay was used to detect and differentiate poxviruses in skin biopsy and skin scab samples from four cattle, one sheep, and one goat. Molecular characterization of capripoxviruses and parapoxviruses was undertaken by sequence analysis of RPO30 and GPCR genes.ResultsThe HRM assay revealed lumpy skin disease virus (LSDV) in three cattle samples, pseudocowpox virus (PCPV) in one cattle sample, and orf virus (ORFV) in one goat and one sheep sample. The phylogenetic analyses, based on the RPO30 and GPCR multiple sequence alignments showed that the LSDV sequences of Botswana were similar to common LSDV field isolates encountered in Africa, Asia, and Europe. The Botswana PCPV presented unique features and clustered between camel and cattle PCPV isolates. The Botswana ORFV sequence isolated from goat differed from the ORFV sequence isolated from sheep.ConclusionsThis study is the first report on the genetic characterization of poxvirus diseases circulating in cattle, goats, and sheep in Botswana. It shows the importance of molecular methods to differentially diagnose poxvirus diseases of ruminants.

Lumpy skin disease (LSD) is an infectious economically important viral disease of cattle. Lumpy skin disease virus (LSDV) is still circulating in Egypt, despite the annual mass vaccination with sheep pox virus vaccine recommended by the Egyptian authorities. This study was carried out on two farms of pregnant Holstein cows vaccinated with Bovivax LSD-N® Vaccine (farm I) and Servac Capri-C vaccine® (farm II). After 40-60 days post-vaccination, mild clinical signs were detected in 3% of cows on the farm I whereas a more severe clinical infection was developed in 40% of cows on farm II. LSDV was isolated on the chorioallantoic membrane (CAM) of 11 days old embryonated chicken egg (ECEs) and Madin Darby bovine kidney (MDBK) tissue culture. LSDV was identified in collected skin tissues by Transmission electron microscopy (TEM) and histopathological examination. Finally, LSDV was confirmed by polymerase chain reaction (PCR) amplification at 192 base pair (bp) of the P32 gene and two samples were selected for DNA sequencing. LSDV developed characteristic pock lesions in inoculated ECEs. MDBK cell culture developed a prominent LSDV cytopathic effect at the 3rd passage. Viral particles were detected in the cytoplasm of both epidermal cells and dermal macrophages by TEM. Histopathological examination revealed different lesions correlated withLSDV infection age. LSDV was confirmed in all tested samples by PCR. Our strains (Dakahlia- 2020-1 and Dakahlia- 2020-2) were closely related to other Egyptian LSDV strains on GenBank with 98.2%-100% identity. The present study proved conclusive evidence that the live attenuated sheep poxvirus vaccine poorly protects Egyptian cattle against LSDV, while the LSDV Neethling strain vaccine gave a promising and sufficient protection rate

Background and Aim:Lumpy skin disease (LSD) and sheep pox are economically important Capripoxvirus-induced diseases of cattle and sheep, respectively. Despite the extensive vaccination program adopted by Egyptian veterinary authorities, LSD and sheep pox are still prevalent and spread throughout the whole country. The current study was designed for molecular characterization and phylogenetic analysis of LSD virus (LSDV) and Sheep pox virus (SPPV) recovered from field cases in Egypt along with vaccinal strains to assess their genetic relatedness.Materials and Methods:Skin biopsies were collected from naturally infected cases of LSD in Ismailia (n=3 farms) and Beni-Suef (n=2 farms) Governorates and sheep pox in Beni-Suef (n=1 flock). Virus isolation was carried out on primary ovine fetal kidney and heart cell cultures. DNA was extracted from infected materials (skin lesions, infected cell cultures) as well as LSDV Neethling vaccine strain and Romanian SPPV vaccine strain. Polymerase chain reaction was performed using oligonucleotide primers targeting the entire open reading frame of G protein-coupled receptors (GPCR) gene and gene sequences were analyzed.Results:Virus isolation on primary ovine fetal kidney and heart cell culture revealed a cytopathic effect at the third passage characterized by rounding of infected cells and margination of nuclear chromatin. Comparative sequence analysis of GPCR gene revealed that Egyptian LSDV isolated from Ismailia and Beni-Suef shared 99:100% nucleotide and amino acid (AA) identities with each other. In comparison to the vaccinal strains, Egyptian LSDV isolates shared 98:99 nucleotide and AA identities with LSDV Neethling vaccine strain and 93:94% with SPPV Romanian vaccine strain. No differences at the nucleotide or AAs were observed between the SPPV vaccine and virulent strains (100% identity). Phylogenetic analyses revealed that LSDV Neethling vaccine strain is more related to field Egyptian LSDV and clustered within the LSDV group while Romanian SPPV vaccine strain clustered in a separate clade with SPPV field isolates.Conclusion:Comparative sequencing and phylogenetic analyses of the GPCR gene reveal a minimal genetic variation between LSDV field isolates from different locations and a close relationship between virulent field strains and homologous vaccines.

Lumpy skin disease virus (LSDV) is a significant threat to cattle, particularly in countries like Thailand, where outbreaks have necessitated the importation of diagnostic kits and vaccines. This study aimed to evaluate several recombinant extracellular enveloped virion (EEV) protein candidates, including F13L, A33R, A34R, and B5R, for their potential use in serological detection assays for LSDV specific antibodies in cattle. Given the challenges associated with LSDV research, such as its classification as a Class III biological agent in Thailand, gene synthesis was employed to produce these proteins. The recombinant proteins were expressed in a prokaryotic system and analyzed using SDS-PAGE and Western blotting. Among the candidates, F13L demonstrated the highest correlation with the results from a commercially available and validated ELISA, yielding 85.7%, and 75% positive for the infected and vaccinated groups, respectively, identifying it a promising candidate for serosurveillance activities during active LSDV outbreaks. Sequence analysis confirmed a 100% match between the F13L designed from the Neethling type strain 2490 and various Thai LSDV strains from the 2021 outbreaks, underscoring its potential as a conserved diagnostic marker. The availability of recombinant F13L and its reactivity with cattle sera from LSDV infected or vaccinated animals, demonstrated in this study, suggests it could also serve as a potential candidate for vaccine development. The study concludes that recombinant F13L shows great promise for the development of LSDV serological assays, though further optimization and validation are necessary to harness its diagnostic potential.

BackgroundLumpy Skin Disease (LSD) is endemic in sub-Saharan countries and is currently a global threat to the cattle industry. Information on the circulating Capripoxvirus lumpyskinpox, formerly known as Lumpy Skin Disease Virus (LSDV), and other poxviruses infecting cattle is very scant in Tanzania. The current study aimed to confirm and characterize LSDV and other poxviruses infecting cattle, from LSD suspected outbreaks in Tanzania.MethodsA total of 24 samples were collected from four LSD suspected outbreaks reported in Tanzania between February and May 2023. Samples were screened for LSDV genome by real-time PCR and then subjected to a high-resolution multiplex melting (HRM) assay where 10 samples were positive for Capripoxvirus (CaPV) and one sample was Parapoxvirus (PPV) positive. Four LSDV genes; RPO30, GPCR, EEV glycoprotein and B22R and the partial B2L gene of PPVs were analyzed.ResultsAll targeted LSDV genes from the Tanzanian isolates showed 100% similarity and isolates clustered with commonly circulating LSDV field isolates. Furthermore, the single nucleotide polymorphism (SNP) at position 240 (A-> G) of the EEV gene differentiates the Tanzanian LSDVs from the group of ancient Kenyan LSDV isolates while the B22R sequences of the Tanzanian LSDV isolates differed from the LSDV Neethling and LSDV KSGP-0240 derived vaccines. Sequence analysis of the partial B2L gene of the Tanzanian parapoxvirus bovinestomatitis, formerly known as Bovine papular stomatitis virus (BPSV) showed a different BPSV strain circulating compared to publicly available sequences.ConclusionThese findings confirm the presence of LSDV in Tanzania, which suggesting the need for establishing an effective control program and continuous monitoring. The presence of a typical profile for Tanzania BPSV is an indication that, although never reported before, BPSV is established in the country therefore this virus should be included in the differential diagnosis of LSDV.

Detection of lumpy skin disease (LSD) virus and distribution of blood-sucking insects as potential vector in Indonesia.

Lumpy Skin Disease (LSD) is a new disease of cattle in Bangladesh. It is endemic in Africa but through the last few years disease beings to spread to other countries of the world. The disease was widely spreaded in the many other countries in Asia and some parts of Europe. In Bangladesh, the disease was first time detected in April 2019, in southern part and then continued to spread all over the country.The disease caused enormous economic losses causing cutaneous and internal lesions, affecting milk production, hide quality and in some cases death of infected animal. LSD suspected samples were collected from different areas of the country during the period from July 2019 to January 2020. In this study, a total of 36 clinically suspected LSD samples of skin crustnodules, pus and ocular discharge were collected. Samples were examined by the published PCR protocol for LSD virus, GPV and SPV. Around 78% samples were found positive for LSD virus in PCR test. LSD virus was also identified from pus and ocular discharge of infected cattle. The virus can grow in the lamb testicular cell and clinically the disease is characterized by distinctive nodular lesions mostly on the skin of the affected animals.The results indicated that the LSD virus is circulating in the outbreak are as and is an emerging transboundary cattle disease in Bangladesh. Bang. J. Livs. Res. Vol. 26 (1&2), 2019: P. 15-20

Lumpy skin disease (LSD), an economically significant disease in cattle caused by lumpy skin disease virus (LSDV), is endemic to nearly all of Africa. Since 2012, LSDV has emerged as a significant epizootic pathogen given its rapid spread into new geographical locations outside Africa, including the Middle East, Eastern Europe, and Asia. To assess the genetic diversity of LSDVs in East Africa, we sequenced and analyzed the RPO30 and GPCR genes of LSDV in twenty-two archive samples collected in Ethiopia, Kenya, and Sudan before the appearance of LSD in the Middle East and its incursion into Europe. We compared them to publicly available sequences of LSDVs from the same region and those collected elsewhere. The results showed that the East African field isolates in this study were remarkably similar to each other and to previously sequenced field isolates of LSDV for the RPO30 and GPCR genes. The only exception was LSDV Embu/B338/2011, a field virus collected in Kenya, which displayed mixed features between the LSDV Neethling vaccine and field isolates. LSDV Embu/B338/2011 had the same 12-nucleotide insertion found in LSDV Neethling and KS-1 vaccines. Further analysis of the partial EEV glycoprotein, B22R, RNA helicase, virion core protein, NTPase, and N1R/p28-like protein genes showed that LSDV Embu/B338/2011 differs from previously described LSDV variants carrying the 12-nucleotide insertion in the GPCR gene. These findings highlight the importance of the constant monitoring of genetic variation among LSDV isolates.

Lumpy skin disease virus (LSDV) is a vector-transmitted poxvirus that causes disease in cattle. Vector species involved in LSDV transmission and their ability to acquire and transmit the virus are poorly characterized. Using a highly representative bovine experimental model of lumpy skin disease, we fed four model vector species (Aedes aegypti, Culex quinquefasciatus, Stomoxys calcitrans, and Culicoides nubeculosus) on LSDV-inoculated cattle in order to examine their acquisition and retention of LSDV. Subclinical disease was a more common outcome than clinical disease in the inoculated cattle. Importantly, the probability of vectors acquiring LSDV from a subclinical animal (0.006) was very low compared with that from a clinical animal (0.23), meaning an insect feeding on a subclinical animal was 97% less likely to acquire LSDV than one feeding on a clinical animal. All four potential vector species studied acquired LSDV from the host at a similar rate, but Aedes aegypti and Stomoxys calcitrans retained the virus for a longer time, up to 8 days. There was no evidence of virus replication in the vector, consistent with mechanical rather than biological transmission. The parameters obtained in this study were combined with data from studies of LSDV transmission and vector life history parameters to determine the basic reproduction number of LSDV in cattle mediated by each of the model species. This reproduction number was highest for Stomoxys calcitrans (19.1), followed by C. nubeculosus (7.1) and Ae. aegypti (2.4), indicating that these three species are potentially efficient transmitters of LSDV; this information can be used to inform LSD control programs.IMPORTANCE Lumpy skin disease virus (LSDV) causes a severe systemic disease characterized by cutaneous nodules in cattle. LSDV is a rapidly emerging pathogen, having spread since 2012 into Europe and Russia and across Asia. The vector-borne nature of LSDV transmission is believed to have promoted this rapid geographic spread of the virus; however, a lack of quantitative evidence about LSDV transmission has hampered effective control of the disease during the current epidemic. Our research shows subclinical cattle play little part in virus transmission relative to clinical cattle and reveals a low probability of virus acquisition by insects at the preclinical stage. We have also calculated the reproductive number of different insect species, therefore identifying efficient transmitters of LSDV. This information is of utmost importance, as it will help to define epidemiological control measures during LSDV epidemics and of particular consequence in resource-poor regions where LSD vaccination may be less than adequate.

Simple SummaryLumpy skin disease is a significant viral infection affecting cattle, and it is characterized by skin nodules, fever, and potentially fatal outcomes, all of which contribute to substantial economic losses in the livestock industry. This review synthesizes existing understanding of the molecular mechanisms employed by the Lumpy Skin Disease Virus (LSDV), focusing on its ability to enter host cells, replicate, and evade immune defenses. Through comparative analysis with vaccinia virus, a well-characterized relative, we elucidate both conserved and unique strategies that enhance viral survival and transmission. The findings demonstrate that LSDV encodes multiple proteins capable of disrupting host immunity, thereby promoting infection. These insights deepen our comprehension of viral pathogenesis and provide a foundation for developing improved vaccines and control measures aimed at mitigating outbreaks, enhancing animal health, and supporting sustainable agricultural economies.Lumpy Skin Disease Virus (LSDV), a member of the poxvirus family, represents a significant threat to global cattle industries. This review presents an analysis of LSDV-encoded proteins and their interactions with host systems, elucidating the molecular mechanisms governing viral life cycle progression and immune evasion strategies. We provide detailed characterization of the complex architecture of LSDV virions, including Intracellular Mature Virus (IMV), Extracellular Enveloped Virus (EEV), lateral bodies, and the core components, while summarizing the crucial functions of viral proteins throughout various stages of infection—entry, replication, transcription, translation, assembly, and egress. Particular attention is given to the immunomodulatory strategies employed by LSDV to subvert both innate and adaptive immune responses. These mechanisms encompass molecular mimicry of cytokines and chemokines, interference with antigen presentation pathways, inhibition of key immune signaling cascades, and modulation of apoptosis and autophagy processes. Through comparative analysis with homologs from related poxviruses, especially vaccinia virus, we highlight both evolutionarily conserved functions and potential unique adaptations in LSDV proteins. This review further identifies critical knowledge gaps in current understanding and proposes promising research directions. We emphasize that integrating multi-omics approaches with structural biology will be essential for advancing our understanding of LSDV pathogenesis and for developing novel preventive and therapeutic strategies against this important animal pathogen.

Lumpy skin disease (LSD) is a severe infectious, emerging transboundary disease of cattle, caused by a Pox family DNA virus. Lumpy skin disease virus (LSDV) infection is associated with a febrile response followed by emergence of widespread dermal nodules. In addition to the skin, LSDV resides in multiple internal organs and can be isolated from the blood of infected cattle. LSDV is suggested to be mechanically transmitted by biting arthropods. Live attenuated vaccines are commonly used to control disease and its spread. We have characterized the tropism, replication, and dissemination of a LSDV field isolate and of an attenuated vaccine strain usingin vitrosystems. To follow virus infection and dissemination in living cells, we have generated recombinant viruses expressing green fluorescent protein (GFP) under a synthetic viral promoter. Recombinant, GFP-expressing, LSDVs demonstrated similar replication kinetics to their corresponding parental LSDV strains in a bovine kidney cell line (MDBK). We further demonstrated that LSDV-GFP productively replicated in a bovine macrophage cell line and in primary bovine foreskin cells with no apparent differences between the field isolate and the vaccine strain. When bovine peripheral blood mononuclear cells (PBMCs) were infected with either LSDV recombinant strain, we observed specific viral driven GFP fluorescence as well as significant viral gene expression. However, infected PBMCs failed to support substantial viral DNA replication and release of infectious progeny. Subsequent analysis of the anti-viral response revealed that heat treated (HT) LSDV induced the expression of interferon- stimulated genes (ISGs) in PBMCs, but this response was suppressed by infectious viruses. Finally, we show that despite failed replication, LSDV infected PBMCs transmitted the virus to recipient co-cultured MDBK cells and produced infectious foci, suggesting a potential role of PBMCs in LSDV dissemination.HighlightsVirulent and attenuated LSDV productively replicated in bovine kidney and bovine macrophage cell lines as well as in primary fibroblasts.Adherent white blood cells were susceptible to LSDV field and attenuated vaccine infection.LSDV showed active viral transcription in PBMCs yet no significant viral genome replication or production of infectious progeny.PBMCs infected with heat-treated LSDV but not with fully infectious viruses upregulated ISGs’ RNA.PBMCs transmitted and disseminated LSDV to contacting permissive cells.

Lumpy skin disease (LSD) is the most fatal viral disease that causes huge economic losses in Cattle farming. The study aims to assess the knowledge, awareness, and practices of dairy farmers in Palampur Tehsil regarding LSD, identify constraints in adopting control measures, and inform effective disease management strategies. The study was conducted from January to June 2025 among 150 farmers from five panchayats (Andreta, Saliana, Sungal, Arla Khas and Paror) in Palampur Tehsil, Kangra district, Himachal Pradesh, India. The survey study on Lumpy Skin Disease in cattle involves systematic data collection and analysis to understand its prevalence, risk factors, and impact on the affected cattle population. The results on the awareness of LSD among dairy farmers revealed that overall, farmers exhibited a neutral or uncertain attitude toward the disease. There was no significant difference in attitude between farmers who had previously encountered lumpy skin disease and those who had not. However, participation in seminars, group discussions, and lectures on lumpy skin disease showed a positive correlation with farmers' attitudes and awareness. Knowledge about the symptoms, modes of transmission, first aid, and preventive and control measures was generally found to be at a high or medium. It is reported that 43.33% dairy farmers exhibited a high level of awareness about lumpy skin disease, while 32.66% and 24% of farmers have medium and low awareness of the disease. Media exposure, interpersonal communication channels, seminars, and workshops attended had a significant role in the general awareness of lumpy skin disease. The majority of the farmers used mobile phones as a source of information (41.33%), other sources are newspaper (33.33%), television (32.66%), and radio (22.66%). The overall distribution of dairy farmers based on awareness of first aid and disease management was also studied, and it is worth stating that the majority of dairy farmers had high awareness of first aid and disease management, i.e. 42.66%, while 36% and 21.33% of farmers had medium and low awareness about disease management. Regarding constraints faced by farmers in adopting control measures, most felt constraints were decreased milk costs during the pandemic (53.33%) and reduction in the production & quality of milk in infected cattle (49.33%).

Lumpy skin disease (LSD) is a vector-borne infection caused by the poxvirus lumpy skin disease virus (LSDV) and is a serious disease of cattle, water buffalo, and banteng. While the disease has never occurred in Australia, it is regarded as a growing threat to the Australian cattle industry as there is on-going spread of the disease throughout Asia. The development of geospatial decision support tools, such as spatial epidemiological modelling, may assist in assessing areas at greater risk of this threat. To guide the design of disease modelling approaches to support future risk-based surveillance, existing LSDV epidemiological models need to be evaluated. In this study, we performed a literature review to evaluate existing LSDV epidemiological models, identify key risk factors for introduction and spread of LSDV, and consider previously adopted control strategies. The PRISMA guidelines were used to establish the processes for article selection and information extraction, and the PICO process was used to formulate search terms. From studies that met our inclusion criteria, we extracted information on LSDV epidemiological model structure and parameterisation, risk factors for LSDV transmission and spread, and biosecurity control strategies. The literature search retrieved a total of 402 articles from four databases, of which 68 were identified for inclusion in this review following screening. Of the 68 articles reviewed, 47 explored risk factors associated with LSDV transmission and spread, four explored risk factors of LSDV introduction, four explored existing surveillance strategies in LSD-free countries, and 14 presented epidemiological models. Our findings indicate that there are various risk factors for LSDV transmission in LSD endemic countries, including long-distance airborne movement of infected vectors such as stable flies and cattle movement between countries over land borders. Key risk factors for LSDV spread in LSD endemic countries include physical environmental characteristics, weather conditions, and population distributions of livestock and vectors. Our results indicate that while a variety of modelling studies have been conducted, the majority of studies experimentally explored LSD transmission mechanisms in vectors and cattle. Spatial and spatio-temporal models have primarily been developed for LSD endemic countries and focus on the spread of the disease in terms of environmental factors in relation to previous LSD events. There were very few studies on LSD-free countries, and these only focussed on risk of LSD introduction through specific entry pathways. This review did not identify any literature exploring the risk of spread of LSDV following introduction in LSD-free countries or geospatial modelling of the suitability of LSD-free countries for LSDV incursions. In conjunction with the risk parameters and models described in the identified literature, there is need to consider a wide range of risk factors specific to Australia to inform the design of risk-based surveillance for LSD in Australia.

Lumpy skin disease virus (LSDV) causes lumpy skin disease (LSD), a highly contagious cattle disease that leads to substantial economic losses to the global cattle industry. Currently, it is imperative to further elucidate its biological characteristics and analyze the global epidemiological dynamics. In this study, two isolates of LSDV with genetic recombination were identified in Northern and Eastern China, which demonstrated broad host cell entry ability. Through electron microscopy, we further revealed its morphogenetic characteristics across its replication cycle for the first time. Viral particles sequentially formed crescent membranes, nucleoids, and lateral bodies, and ultimately developed into four types of mature virions: intracellular mature virions (IMVs), intracellular enveloped virions (IEVs), cell-associated enveloped virions (CEVs), and extracellular enveloped virions (EEVs). Whole-genome phylogenetic analysis revealed that both isolates belonged to Clade R4. Based on the global reference strains, our integrated analysis for temporal and geographical information revealed that LSDV has progressively expanded its endemic range, particularly in Asia, where recombinant Clade R4 strains have recently emerged as the predominant epidemic strains. Strikingly, recombination analysis detected a limited number of recombination signals between the LSDV isolates and goatpox virus (GTPV) or sheeppox virus (SPPV) strains, suggesting that the possibility of recombination between heterologous vaccines and LSDV cannot be fully excluded. These data may provide important information for prevention and control of LSD global outbreaks.

Lumpy skin disease (LSD) is an endemic infectious disease of cattle in Egypt. This survey aimed to define the prevalence of clinical and sub-clinical LSD virus (LSDV) infection among cattle and investigate their contact with water buffaloes (Bubalus bubalis) in order to improve the understanding of LSD epidemiology. Cattle and buffalo were examined owing to the appearance of skin lesions. Because clinical signs were consistent with LSDV infection, samples from cattle in a non-grazing dairy farm (n = 450) were submitted for LSDV testing together with those from the in-contact buffaloes (n = 100). Results revealed that the intra-herd percentage of cattle infected with LSDV varied with the detection method. This ranged from 22.4% to 65.4% by virus isolation (VI) and polymerase chain reaction (PCR), respectively, in clinical cattle samples, compared to 0% and 10% by VI and PCR in non-clinical cases. Using the neutralising index (NI), LSDV antibodies were found in 100% (n = 100) of the tested cow’s sera (NI = > 2.0 and ≥ 3.0), whereas buffalo’s sera (n = 34) displayed little increase in antibody level (NI ≥ 1.5). None of the buffalo were positive for LSDV by VI and PCR. In addition, there were no significant differences in LSD prevalence among the cattle with regard to age and sex. In conclusion, the occurrence of LSD in cattle warrants a further epidemiological study of the spread of the disease in the area and adoption of control and prevention strategies. In addition, the PCR assay was confirmed to be useful in the diagnosis of LSDV and for wider epidemiological studies.