Evidence of lumpy skin disease virus over-wintering by transstadial persistence in Amblyomma hebraeum and transovarial persistence in Rhipicephalus decoloratus ticks.

Lumpy skin disease is a debilitating cattle disease caused by the lumpy skin disease virus (LSDV), belonging to the genus Capripoxvirus. Epidemics of the disease usually occur in summer, when insect activity is high. Limited information is available on how LSDV persists during inter-epidemic periods. Transmission of LSDV by mosquitoes such as Aedes aegypti has been shown to be mechanical, there is no carrier state in cattle and the role of wildlife in the epidemiology of the disease seems to be of minor importance. Recent studies in ticks have shown transstadial persistence of LSDV in Rhipicephalus appendiculatus and Amblyomma hebraeum as well as transovarial persistence of the virus in Rhipicephalus decoloratus, R. appendiculatus and A. hebraeum. The over-wintering of ticks off the host as part of their life cycles is well known: A. hebraeum and R. appendiculatus over-winter, for example, on the ground as engorged nymphs/unfed (emergent) adults while R. decoloratus over-winters on the ground as engorged females. In this study, transstadial and transovarial persistence of LSDV from experimentally infected A. hebraeum nymphs and R. decoloratus females after exposure to cold temperatures of 5 °C at night and 20 °C during the day for 2 months was reported. This observation suggests possible over-wintering of the virus in these tick species.

Mechanical transmission of lumpy skin disease virus by Rhipicephalus appendiculatus male ticks.

Lumpy skin disease (LSD) is an economically important, acute or sub-acute, viral disease of cattle that occurs across Africa and in the Middle East. The aim of this study was to investigate if lumpy skin disease virus (LSDV) can be transmitted mechanically by African brown ear ticks (Rhipicephalus appendiculatus Neum.). Laboratory-bred R. appendiculatus males were fed on experimentally infected viraemic 'donor' cattle. Partially fed male ticks were then transferred to feed on an uninfected 'recipient' cow. The recipient animal became viraemic, showed mild clinical signs of LSD and seroconverted. Additionally, R. appendiculatus males were found to transmit LSDV through feeding on skin lacking visible lesions, demonstrating that viraemic animals without lesions at the feeding site of ticks may be a source of infection. This is the first time that transmission of poxviruses by a tick species has been demonstrated and the importance of this mode of transmission in the spread of LSDV in endemic settings is discussed.

Detection of lumpy skin disease virus in saliva of ticks fed on lumpy skin disease virus-infected cattle.

Lumpy skin disease is an economically important disease of cattle that is caused by the lumpy skin disease virus (LSDV), which belongs to the genus Capripoxvirus. It is endemic in Africa and outbreaks have also been reported in the Middle-East. Transmission has mostly been associated with blood-feeding insects but recently, the authors have demonstrated mechanical transmission by Rhipicephalus appendiculatus as well as mechanical/intrastadial and transstadial transmission by Amblyomma hebraeum. Saliva is the medium of transmission of pathogens transmitted by biting arthropods and, simultaneously, it potentiates infection in the vertebrate host. This study aimed to detect LSDV in saliva of A. hebraeum and R. appendiculatus adult ticks fed, as nymphs or as adults, on LSDV-infected animals, thereby also demonstrating transstadial or mechanical/intrastadial passage of the virus in these ticks. Saliva samples were tested for LSDV by real-time PCR and virus isolation. Supernatants obtained from virus isolation were further tested by real-time PCR to confirm that the cytopathic effects observed were due to LSDV. Lumpy skin disease virus was detected, for the first time, in saliva samples of both A. hebraeum and R. appendiculatus ticks. At the same time, mechanical/intrastadial and transstadial passage of the virus was demonstrated and confirmed in R. appendiculatus and A. hebraeum.

Goatpox virus (G20-LKV) vaccine strain elicits a protective response in cattle against lumpy skin disease at challenge with lumpy skin disease virulent field strain in a comparative study.

In this comparative study, we examine the safety of the sheeppox (SPP) and goatpox (GTP) vaccines and the protective response of these vaccines in cattle against a virulent lumpy skin disease (LSD) field strain. The vaccine safety was tested in rabbits, mice and cattle using ten times recommended dose. In the safety trial, none of the vaccinated animals showed any deviation from physiological norms or fever, inappetence or local/ generalized skin reactions. In the challenge trial, both SPP and GTP vaccine groups developed virus-neutralizing antibodies with an average titre of 2.1 log2 at 21 days post-vaccination. No significant difference in seroconversion was found in cattle vaccinated with SPP and GTP vaccines (P ≥ 0.05). When challenged with a virulent LSD field strain, one animal vaccinated with the SPP Niskhi vaccine strain showed typical LSD skin lesions at the injection sites of different dilutions of the challenge virus. All animals vaccinated with GTP G20-LKV vaccine strain showed full protection. After infection with the challenge virus, unvaccinated fully susceptible control cattle showed characteristic clinical signs of LSD. The average protective index for SPP and GTP vaccine groups was 5.3 ± 1.42 and 5.9 ± 0.00, respectively.

Field observations and experiences gained from the implementation of control measures against lumpy skin disease in South-East Europe between 2015 and 2017.

The first epidemics of lumpy skin disease (LSD) reported in Europe in 2015 severely affected the cattle farming sector in several Balkan countries. After the first incursion into Greece in 2015, the disease quickly spread across the Balkan region with over 7000 outbreaks reported by the end of 2016. Thanks to a coordinated regional control and eradication policy, the spread of the disease was halted by the end of 2017. Regional large-scale vaccination campaign with effective homologous vaccines and high vaccination coverage revealed to be essential for the successful control the disease, supported by other measures such as early detection of outbreaks, total or partial stamping out and restrictions on cattle movements. The aim of this paper is to discuss the field observations, challenges and lessons learnt while dealing with the first LSD epidemics in Europe. The cross-border collaboration by the veterinary authorities of all affected countries, coordinated by the European Commission and the technical support provided by many other international organizations played a fundamental role in stopping the spread of a disease that otherwise could have expanded further to the European territory causing a large damage to the whole European cattle farming industry. The experience obtained during the control of LSD epidemics indicates that in the future LSD spread can be effectively halted, provided that appropriate surveillance plans and vigilance remains in place in the areas at risk of re-incursion, especially those bordering endemic countries.

Transmission of lumpy skin disease virus: A short review.

Lumpy skin disease (LSD) is a viral transboundary disease endemic throughout Africa and of high economic importance that affects cattle and domestic water buffaloes. Since 2012, the disease has spread rapidly and widely throughout the Middle Eastern and Balkan regions, southern Caucasus and parts of the Russian Federation. Before vaccination campaigns took their full effect, the disease continued spreading from region to region, mainly showing seasonal patterns despite implementing control and eradication measures. The disease is capable of appearing several hundred kilometers away from initial (focal) outbreak sites within a short time period. These incursions have triggered a long-awaited renewed scientific interest in LSD resulting in the initiation of novel research into broad aspects of the disease, including epidemiology, modes of transmission and associated risk factors. Long-distance dispersal of LSDV seems to occur via the movement of infected animals, but distinct seasonal patterns indicate that arthropod-borne transmission is most likely responsible for the swift and aggressive short-distance spread of the disease. Elucidating the mechanisms of transmission of LSDV will enable the development of more targeted and effective actions for containment and eradication of the virus. The mode of vector-borne transmission of the disease is most likely mechanical, but there is no clear-cut evidence to confirm or disprove this assumption. To date, the most likely vectors for LSDV transmission are blood-sucking arthropods such as stable flies (Stomoxys calcitrans), mosquitoes (Aedes aegypti), and hard ticks (Rhipicephalus and Amblyomma species). New evidence suggests that the ubiquitous, synanthropic house fly, Musca domestica, may also play a role in LSDV transmission, but this has not yet been tested in a clinical setting. The aim of this review is to compile and discuss the earlier as well as the most recent research data on the transmission of LSDV.

Experimental evidence of mechanical lumpy skin disease virus transmission by Stomoxys calcitrans biting flies and Haematopota spp. horseflies.

Lumpy skin disease (LSD) is a devastating disease of cattle characterized by fever, nodules on the skin, lymphadenopathy and milk drop. Several haematophagous arthropod species like dipterans and ticks are suspected to play a role in the transmission of LSDV. Few conclusive data are however available on the importance of biting flies and horseflies as potential vectors in LSDV transmission. Therefore an in vivo transmission study was carried out to investigate possible LSDV transmission by Stomoxys calcitrans biting flies and Haematopota spp. horseflies from experimentally infected viraemic donor bulls to acceptor bulls. LSDV transmission by Stomoxys calcitrans was evidenced in 3 independent experiments, LSDV transmission by Haematopota spp. was shown in one experiment. Evidence of LSD was supported by induction of nodules and virus detection in the blood of acceptor animals. Our results are supportive for a mechanical transmission of the virus by these vectors.

Review: Capripoxvirus Diseases: Current Status and Opportunities for Control.

Lumpy skin disease, sheeppox and goatpox are high-impact diseases of domestic ruminants with a devastating effect on cattle, sheep and goat farming industries in endemic regions. In this article, we review the current geographical distribution, economic impact of an outbreak, epidemiology, transmission and immunity of capripoxvirus. The special focus of the article is to scrutinize the use of currently available vaccines to investigate the resource needs and challenges that will have to be overcome to improve disease control and eradication, and progress on the development of safer and more effective vaccines. In addition, field evaluation of the efficacy of the vaccines and the genomic database available for poxviruses are discussed.

Detection of Capripoxvirus DNA Using a Field-Ready Nucleic Acid Extraction and Real-Time PCR Platform.

Capripoxviruses, comprising sheep pox virus, goat pox virus and lumpy skin disease virus cause serious diseases of domesticated ruminants, notifiable to The World Organization for Animal Health. This report describes the evaluation of a mobile diagnostic system (Enigma Field Laboratory) that performs automated sequential steps for nucleic acid extraction and real-time PCR to detect capripoxvirus DNA within laboratory and endemic field settings. To prepare stable reagents that could be deployed into field settings, lyophilized reagents were used that employed an established diagnostic PCR assay. These stabilized reagents demonstrated an analytical sensitivity that was equivalent, or greater than the established laboratory-based PCR test which utilizes wet reagents, and the limit of detection for the complete assay pipeline was approximately one log10 more sensitive than the laboratory-based PCR assay. Concordant results were generated when the mobile PCR system was compared to the laboratory-based PCR using samples collected from Africa, Asia and Europe (n = 10) and experimental studies (n = 9) representing clinical cases of sheep pox, goat pox and lumpy skin disease. Furthermore, this mobile assay reported positive results in situ using specimens that were collected from a dairy cow in Morogoro, Tanzania, which was exhibiting clinical signs of lumpy skin disease. These data support the use of mobile PCR systems for the rapid and sensitive detection of capripoxvirus DNA in endemic field settings.

Adverse Reactions to Field Vaccination Against Lumpy Skin Disease in Jordan.

Lumpy skin disease (LSD) is an emerging disease in the Middle East region and has been recently reported in Jordan. The aim of this study was to investigate the adverse reactions that were reported after vaccine administration. Geographical areas enrolled in the study were free of the disease and away from the outbreak governorate. Sixty-three dairy cattle farms, with a total of 19,539 animals, were included in the study. Of those, 56 farms reported adverse clinical signs after vaccine administration. The duration between vaccine administration and appearance of adverse clinical signs ranged from 1 to 20 days (Mean = 10.3, SD ± 3.9). Clinical signs were similar to those observed with natural cases of lumpy skin disease. These were mainly fever, decreased feed intake, decreased milk production and variable sized cutaneous nodules (a few millimetres to around 2 cm in diameter) that could be seen anywhere on the body (head, neck, trunk, perineum), udder, and/or teats. Nodules were raised and firm initially and then formed dry scabs that could be peeled off the skin. The characteristic deep 'sit fast' appearance was rarely seen and most lesions were superficial. Some cattle had swollen lymph nodes, while a few pregnant animals aborted. The percentage of affected cattle ranged from 0.3 to 25% (Mean = 8, SD ± 5.1). Fever, decreased feed intake, and decreased milk production were seen in 83.9, 85.7, and 94.6% in cattle on the affected farms, respectively. All affected cattle displayed skin nodules over their entire bodies, while 33.9 and 7.1% of the affected farms reported nodular lesions present on the udders and teats, respectively. No mortalities were reported due to vaccine adverse reactions. Duration (course) of clinical signs ranged from 3 to 20 days (Mean = 13.7, SD ± 4.1). Two types of LSD vaccines were used by the farmers in this study. The first one was a sheep pox virus (SPPV) vaccine derived from the RM65 isolate [Jovivac, manufactured by Jordan Bioindustries Centre (JOVAC)] and the other an unlabelled one, which was later identified using PCR as a strain of lumpy skin disease virus (LSDV). Blood and skin samples collected from cattle vaccinated with the LSDV vaccine were positive for LSDV using both general and species-specific PCR primers, whereas those from cattle vaccinated with the Jovivac vaccine were negative. Adverse reactions observed in cattle after administration of the LSDV vaccine were reported to be more severe than those seen after Jovivac vaccine administration and were comparable with clinical signs observed in natural infections.

Excretion of lumpy skin disease virus in bull semen.

This work was done to establish the incidence and duration of excretion of lumpy skin disease virus (LSDV) in semen of experimentally infected susceptible bulls. Six serologically negative bulls 11-20 months of age were experimentally infected with a virulent field isolate (strain V248/93) of LSDV. Animals were observed for the development of clinical signs, blood was collected until day 90 after infection, and semen was collected every second day until day 18, then twice a week till day 63 and twice a month until three consecutive samples were negative when tested for LSDV by polymerase chain reaction (PCR). An aliquot of each sample which tested positive using PCR was inoculated onto cell monolayers for the recovery of virus. Two bulls developed severe lumpy skin disease (LSD), two bulls showed mild signs and two bulls showed a transient fever only. Multiple samples were positive on PCR from both of the severely affected bulls and one of the mildly affected bulls; between days 10 and 159, days 8 and 132, and days 10 and 21 respectively. Only one sample from each of the other three bulls was positive on PCR. Virus was only isolated from two samples from one of the severely affected bulls and from five semen samples from the other. This study confirmed the excretion of LSDV in bovine semen for prolonged periods, even when obvious clinical signs of the disease were no longer apparent.

The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques.

Lumpy skin disease (LSD) is a disease of cattle, primarily in Africa and Madagascar and rarely in the Middle East. It is caused by a capripoxvirus that belongs to the family Poxviridae. The disease is of economic importance in endemic areas. Effective control of LSD requires accurate and rapid laboratory techniques to confirm a tentative clinical diagnosis. Comparative studies on different diagnostic tests used at different stages of the disease have not been done. The aim of this study was to compare several of these tests. Six seronegative bulls, between 11 and 20 months of age, were infected intravenously and kept in an insect-free facility. The course of the infection was monitored. During a 3-month period blood samples and skin biopsies were collected for virus isolation and polymerase chain reaction (PCR). Skin biopsies were also examined using transmission electron microscopy (TEM). The incubation period in infected animals varied from 4-5 days. The length of the viraemic period did not correlate with the severity of clinical disease. Viraemia was detected from 1-12 days using virus isolation and from 4-11 days using the PCR, which is longer than has previously been reported. Virus was isolated from skin biopsies until Day 39 post infection (p.i.) and PCR could demonstrate viral DNA until Day 92 p.i. Transmission electron microscopy of negatively stained skin biopsies detected LSD virus only in one of the four bulls that developed skin lesions until Day 33 p.i. The PCR was a fast and sensitive method to demonstrate viral DNA in blood and skin samples. It could detect viral nucleic acid in skin lesions 53 days longer than virus isolation. Virus isolation from blood and skin samples was sensitive and reliable, but as a single test it may be too time-consuming to use although this depends on how rapidly the diagnosis must be confirmed. In conclusion, this study showed the PCR to be superior in detecting LSD virus from blood and skin samples. However, virus isolation is still required when the infectivity of the LSD virus is to be determined.

Lumpy skin disease: attempted propagation in tick cell lines and presence of viral DNA in field ticks collected from naturally-infected cattle.

Lumpy skin disease (LSD) is of substantial economic importance for the cattle industry in Africa and the Near and Middle East. Several insect species are thought to transmit the disease mechanically. Recent transmission studies have demonstrated the first evidence for a role of hard (ixodid) ticks as vectors of lumpy skin disease virus (LSDV). The aim of this study was to attempt in vitro growth of the virus in Rhipicephalus spp. tick cell lines and investigate in vivo the presence of the virus in ticks collected from cattle during LSD outbreaks in Egypt and South Africa. No evidence was obtained for replication of LSDV in tick cell lines although the virus was remarkably stable, remaining viable for 35 days at 28°C in tick cell cultures, in growth medium used for tick cells and in phosphate buffered saline. Viral DNA was detected in two-thirds of the 56 field ticks, making this the first report of the presence of potentially virulent LSDV in ticks collected from naturally infected animals.

Evidence of transstadial and mechanical transmission of lumpy skin disease virus by Amblyomma hebraeum ticks.

Lumpy skin disease (LSD) is an economically important disease caused by LSD virus (LSDV), a Capripoxvirus, characterized by fever and circumscribed skin lesions. It is suspected to be transmitted mechanically by biting flies. To assess the vector potential of Amblyomma hebraeum in transmission of LSDV, mechanical/intrastadial and transstadial modes of transmission of the virus by this tick species were investigated. Two cattle were artificially infected as sources (donors) of infection to ticks. Ticks were infected as either nymphs or adults. Male A. hebraeum ticks were partially fed on donor animals and transferred to recipient animals to test for mechanical/intrastadial transmission. Nymphal A. hebraeum were fed to repletion on donor animals. The emergent adult ticks were placed on recipient animals to test for transstadial transmission of the virus. Successful transmission of LSDV infection was determined in recipient animals by monitoring development of clinical signs, testing of blood for the presence of LSDV by real-time PCR, virus isolation and the serum neutralization test. This report provides further evidence of mechanical/intrastadial and, for the first time, transstadial transmission of LSDV by A. hebraeum. These findings implicate A. hebraeum as a possible maintenance host in the epidemiology of the disease.

Review: lumpy skin disease: an emerging threat to Europe, the Middle East and Asia.

Lumpy skin disease (LSD) is an economically devastating emerging viral disease of cattle. Lumpy skin disease is currently endemic in most African countries and has recently spread out of Africa into the Middle East region. In this article, we review the putative mechanisms of spread of LSD into the Middle East and the risks of further spread into Turkey, Europe and Asia. We also review the latest findings on the epidemiology of LSD, its mechanisms of transmission, the potential role of wildlife in its maintenance and spread and the diagnostic tests and control methods currently available.

A potential role for ixodid (hard) tick vectors in the transmission of lumpy skin disease virus in cattle.

Lumpy skin disease (LSD) is an economically important cattle disease. The disease is endemic in many African countries, but outbreaks have also been reported in Madagascar and the Middle East. The aim of this study was to investigate the potential role of ixodid (hard) ticks in the transmission of the disease. Cattle were infected with a virulent, South African field isolate of lumpy skin disease virus (LSDV). Three common African tick species (genera Rhipicephalus, Amblyomma and Rhipicephalus (Boophilus)) in different life cycle stages were fed on the infected animals during the viraemic stage and on skin lesions. Post-feeding, the partially fed male ticks were transferred to the skin of non-infected 'recipient' animals, while females were allowed to lay eggs that were then tested using the polymerase chain reaction (PCR) method and virus isolation. Nymphs were allowed to develop for 2-3 weeks after which time they were tested. The non-infected 'recipient' cattle were closely monitored, both skin and blood samples were tested using PCR and virus isolation, and serum samples were tested by the serum neutralization test. This is the first report showing molecular evidence of potential transmission of LSDV by ixodid ticks. The study showed evidence of transstadial and transovarial transmission of LSDV by R. (B.) decoloratus ticks and mechanical or intrastadial transmission by R. appendiculatus and A. hebraeum ticks.

A description of two outbreaks of capripoxvirus disease in Mongolia.

Mongolia had no reported cases of capripoxvirus disease from 1977 until an outbreak of sheeppox in 2006-2007 and then goatpox in 2008. The two outbreaks occurred in geographically distant areas of Mongolia and, most strikingly, were highly species-specific. The 2006-2007 sheeppox outbreak affected no goats and the 2008 goatpox outbreak affected no sheep despite communal herding. The diseases were diagnosed using the polymerase chain reaction and virus neutralisation test. The P32 gene of the Mongolian sheeppox and goatpox viruses from the recent outbreaks were sequenced and compared with an archived 1967 strain of Goatpox virus from Mongolia. The P32 gene of the 2006-2007 Mongolian Sheeppox virus strain was identical to previously published sheeppox strains. The P32 gene of the 2008 Mongolian Goatpox virus strain was identical to the gene from virus isolated from recent goatpox outbreaks in China and Vietnam. The archived Mongolian Goatpox virus strain was unique.