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In September 2022, more than 50 years after its eradication from Spain, Sheep pox virus was confirmed by laboratory analysis in sheep showing characteristic lesions. This was the start of an outbreak that lasted 9 months and infected 30 farms dispersed over two different areas, Andalusia and Castilla-La Mancha. Early after the initial confirmation, an active surveillance based on clinical inspection with laboratory confirmation of sheep with clinical signs was started in restricted areas. This allowed the confirmation of Sheep pox in 22 out of 28 suspected farms, where limited numbers of sheep with mainly erythema and papules were found, indicative of early detection. Nevertheless, to improve active surveillance and stop the outbreak, clinical inspection was reinforced by laboratory analysis in all inspected farms, even when no clinically diseased sheep were detected. Although more than 35,000 oral swabs from 335 farms were analysed by real-time PCR in pools of five, only two out of six reported outbreaks in this period were detected by laboratory analysis before clinical signs were observed. Furthermore, additional insights were gained from the extensive laboratory surveillance performed on samples collected under field conditions. No evidence of Sheep pox virus infection was found in goats. Oral swabs proved to be the sample of choice for early detection in the absence of scabs and could be tested in pools of five without extensive loss in sensitivity; serology by ELISA was not useful in outbreak detection. Finally, a non-infectious genome of the virus could be detected months after cleaning and disinfection; thus, real-time PCR results should be interpreted with caution in sentinel animals during repopulation. In conclusion, the outbreak of Sheep pox virus in Spain showed that active clinical inspection with laboratory confirmation of clinically diseased sheep via oral swab testing proved a sensitive method for detection of infected farms, providing insights in laboratory surveillance that will be helpful for other countries confronted with Sheep pox outbreaks. [ABSTRACT FROM AUTHOR]

Pest des petits ruminants (PPR) and sheep pox (SP) viruses represent a hazard facing the sheep population. The present work dealt with the preparation of a combined live attenuated vaccine that protects sheep against the two diseases. Successfully, combined Peste des petits ruminants and sheep pox freeze-dried vaccine was proven to sterile and be free from different aerobic and anaerobic bacteria, fungi, and mycoplasma contaminants; safe (as no atypical reaction either local or systemic in mice and sheep and did not affect close contact unvaccinated sheep) and potent (providing vaccinated sheep with high protective specific immunity against both causative viruses) for 6 months (the experimental period) suggesting that such immunity will remain to not less than one year. It could be recommended the use of such a vaccine to control the two diseases using one shot of vaccination saves time, effort, and cost. [ABSTRACT FROM AUTHOR]

In September 2022, more than 50 years after its eradication from Spain, Sheep pox virus was confirmed by laboratory analysis in sheep showing characteristic lesions. This was the start of an outbreak that lasted 9 months and infected 30 farms dispersed over two different areas, Andalusia and Castilla-La Mancha. Early after the initial confirmation, an active surveillance based on clinical inspection with laboratory confirmation of sheep with clinical signs was started in restricted areas. This allowed the confirmation of Sheep pox in 22 out of 28 suspected farms, where limited numbers of sheep with mainly erythema and papules were found, indicative of early detection. Nevertheless, to improve active surveillance and stop the outbreak, clinical inspection was reinforced by laboratory analysis in all inspected farms, even when no clinically diseased sheep were detected. Although more than 35,000 oral swabs from 335 farms were analysed by real-time PCR in pools of five, only two out of six reported outbreaks in this period were detected by laboratory analysis before clinical signs were observed. Furthermore, additional insights were gained from the extensive laboratory surveillance performed on samples collected under field conditions. No evidence of Sheep pox virus infection was found in goats. Oral swabs proved to be the sample of choice for early detection in the absence of scabs and could be tested in pools of five without extensive loss in sensitivity; serology by ELISA was not useful in outbreak detection. Finally, a non-infectious genome of the virus could be detected months after cleaning and disinfection; thus, real-time PCR results should be interpreted with caution in sentinel animals during repopulation. In conclusion, the outbreak of Sheep pox virus in Spain showed that active clinical inspection with laboratory confirmation of clinically diseased sheep via oral swab testing proved a sensitive method for detection of infected farms, providing insights in laboratory surveillance that will be helpful for other countries confronted with Sheep pox outbreaks.

Sheep and goat pox (SGP) disease is a highly contagious and dangerous viral infection, characterized by fever, formation of papular-pustular lesions in the epithelium of the skin and respiratory tract of small ruminants. This disease causes enormous damage to sheep breeding due to the death of animals, forced slaughter, loss of productivity, costs for veterinary and sanitary and quarantine measures. The causative agents are two closely related viruses: sheep pox virus and goat pox virus, belonging to the family Poxviridae, the genus Capripoxvirus. SGP is considered an endemic in Kazakhstan and SGP outbreaks have been registered in a number of regions both in our country and bordering states. The uncontrolled spread of this infection can be associated with colossal economic losses and significant damage of the image of our country as an exporter of lamb. Thus, it is important to monitore the epizootological situation of SGP in the country, control outbreaks of infection, and develop veterinary and sanitary measurements to adequately anticipate of the disease spread. Therefore, the purpose of this study is to perform risk analyses of SGP transmission in Kazakhstan. As a result of the research, the SGP risk factors, epizootological characteristics of this infection in Kazakhstan in the last 10 years, and the districts with high risks regarding the possible occurrence of large outbreaks of SGP were identified and the epizootic visualization map was created. This work provides valuable information on the importance of SGP control and prevention programs in Kazakhstan. [ABSTRACT FROM AUTHOR]

Sheep pox is a highly contagious infectious viral disease (F/Poxviridae, G/Capripoxvirus), still causing economic damage in endemic zones like northern Africa, the Middle East, and many regions in Asia. Nevertheless, the clinical symptoms caused by sheep pox virus (SPPV) is similar to those caused by genetically related viruses, notably goat pox virus (GTPV) and lumpy skin disease virus (LSDV). One of the main objectives of the current study is to characterize the virus; nasal swabs were taken from sheep showing SPPV clinical signs, submitted to molecular sequencing based on the P32 gene marker, and proceeded to genetic comparisons. Phylogenetic results showed that all virus isolate sequences cluster in the SPPV clade, thus distinguishing them from those of GTPV and LSDV. These results give an accurate diagnostic and a clear view of endemic viral strains, guiding veterinary managers in control politics, especially in the choice of a homologous vaccine.

Introduction. Outbreaks of infectious diseases seriously hinder the preservation and increase of the number of small ruminants. Such infections include sheep pox virus (SPPV). According to the OIE data of 2021, SPP outbreaks were registered in countries such as Turkey, Israel, China, Maldives, Mongolia, Thailand, Russia, Algeria, Kenya, and in 2019 in Mangistau and Atyrau regions. In Kazakhstan annually conducts routine immunization of sheep at risk with a live attenuated vaccine produced by RIBSP. Materials and methods. The object of the study was the vaccine strain of NISHI and the virulent strain A of the sheep pox virus. The virus was propagated in Vero cells. To determine the harmlessness and immunogenicity, sheep of the Kazakh fine-wool breed aged from 6 to 12 months were used. Virological, serological and immunobiological methods were used in the study. Results. The results of the adaptation of the NISHI strain of SPPV to the Vero cell line are presented. Five passages in Vero cells resulted to the adaptation of the NISHI strain with the manifestation of a cytopathogenic effect specific to SPPV with a titer of 6.50 lg TCD50/ml. Following immunization, the formation of immunity was observed in animals on day 7 with an average protective titer 1.8 log2, which increased by day 21 to 4.33 log2. Conclusion. It has been established that the NISHI strain of SPPV retains its virological and immunobiological properties during reproduction in a Vero cell line.

Sheep pox (SPP) constitutes a global animal health scourge, despite the numerous efforts targeting the eradication of the disease implemented in affected countries. An efficient control and eradication strategy incorporates the use of live attenuated vaccines, which in turn requires a method for differentiation between vaccinated and infected sheep. The NISKHI live attenuated SPP vaccine (LAV) is abundantly used in Russia, Kazakhstan and other Central Asian countries. This study describes the development and evaluation of a real-time PCR with a high-resolution melting assay, capable of differentiating the NISKHI vaccine virus from circulating virulent field strains. The RNA polymerase subunit RPO132 gene contains a unique single nucleotide polymorphism (SNP) capable of altering the melting curves of amplicons from LAV and virulent field isolates circulating in the region. The melting temperature (Tm) of field isolates ranged from 75.47 °C ± 0.04 to 75.86 °C ± 0.08, while the vaccine strain averaged 76.46 °C ± 0.12. Subsequent evaluation of this assay demonstrated that the recent SPP outbreaks in central Russia may be attributed to virulent field isolates. This robust assay was proven to consistently and differentially detect the NISKHI LAV strain when analyzing clinical samples from affected sheep. [ABSTRACT FROM AUTHOR]

Modeling of potential nosoareals of sheep pox and goat pox that was performed in 2016 revealed a possible trend for aggravation of epizootic situation intensity with regard to these diseases up to 2020, but taking into account the cyclical fluctuations of the situation, it was assumed that the intensity level might reduce in 2017–2020. The highest frequency of reporting these nosounits was characteristic of regions with a higher probability of sheep pox and goat pox occurrence. The retrospective analysis was used to determine the structure, dynamics and properties of the epizootic process of these infectious diseases in the Republic of Tajikistan in 2000–2021. The main causes of sheep pox and goat pox outbreaks in the country were identified. The obtained results confirming the methodological validity of the epizootic situation analysis carried out in 2016 and correctness of the developed models of potential sheep pox and goat pox nosoareals, can become the basis for epizootological prediction of infectious disease risk when the pathogen interacts with susceptible animal population in specific climatic, socio-economic, organizational and managemental conditions. Based on systemic epidemiological analysis of the structure and dynamics of nosoareals, as well as risk assessment of sheep pox and goat pox entry, emergence and distribution, monitoring and establishment of infected and endemic zones, the features and patterns of distribution and occurrence of these diseases’ epizootic process were defined, which confirms the need for a systematic approach to epidemiological surveillance of highly dangerous and economically significant infectious diseases having a trend for transboundary spread, which will facilitate the solution of the problem regarding identification of possible threats and the implementation of veterinary and sanitary measures in case of disease occurrence in the territory of any country to ensure animal disease freedom.

Capripoxviruses include lumpy skin disease virus, sheep pox virus, and goat pox virus, which lead to economic impacts in ruminants. Live, attenuated commercial vaccines can be applied in cattle against lumpy skin disease virus because of cross-protection. Bakırkoy sheep pox vaccine is a live, attenuated vaccine used against lumpy skin disease virus in cattle in Turkey. Continuous cell lines are used in viral vaccine production. This study was performed to the propagation of vaccine virus in the Madin-Darby bovine kidney cells and determination of growth kinetics. At the seventh day post-inoculation, the cells became round and moved away from the surrounding cells and cytopathic effect formation started. After 80% cytopathic effect was formed on the tenth day post-inoculation, virus suspension was harvested and identified as sheep pox virus with real-time polymerase chain reaction analysis and serum neutralization test. The titre of the sheep pox-Madin-Darby bovine kidney vaccine virus was calculated as 105.75/mL TCID50. The highest value was observed on the seventh day but the titre decreased on the ninth day while virus titre of supernatant was detected the highest on the seventh day post-infection and it decreased on the eighth day post-inoculation. The highest titre was reached in the third freezing and thawing and decreased in the fifth thaw. In conclusion, the Madin-Darby bovine kidney cells can be used in sheep pox vaccine production. [ABSTRACT FROM AUTHOR]

Abstract Sheep pox, goat pox, and lumpy skin diseases are economically significant and contagious viral diseases of sheep, goats and cattle, respectively, caused by the genus Capripoxvirus (CaPV) of the family Poxviridae. Currently, CaPV infection of small ruminants (sheep and goats) has been distributed widely and are prevalent in Central Africa, the Middle East, Europe and Asia. This disease poses challenges to food production and distribution, affecting rural livelihoods in most African countries, including Ethiopia. Transmission occurs mainly by direct or indirect contact with infected animals. They cause high morbidity (75-100% in endemic areas) and mortality (10-85%). Additionally, the mortality rate can approach 100% in susceptible animals. Diagnosis largely relies on clinical symptoms, confirmed by laboratory testing using real-time PCR, electron microscopy, virus isolation, serology and histology. Control and eradication of sheep pox virus (SPPV), goat pox virus (GTPV), and lumpy skin disease (LSDV) depend on timely recognition of disease eruption, vector control, and movement restriction. To date, attenuated vaccines originating from KSGPV O-180 strains are effective and widely used in Ethiopia to control CaPV throughout the country. This vaccine strain is clinically safe to control CaPV in small ruminants but not in cattle which may be associated with insufficient vaccination coverage and the production of low-quality vaccines.

As notifiable diseases, lumpy skin disease (LSD), sheep pox (SPP), and goat pox (GTP) are associated with a profound effect on cattle, sheep, and goat farming industries. Development of the ELISA method could effectively facilitate serodiagnosis of the infected animals. This study aimed to develop an ELISA system based on the recombinant full-length and truncated P32 protein (Tr.P32) of goat pox virus. The P32 protein was expressed in Rosetta strain of E. coli using pET24a+ vector and evaluated by SDS-PAGE and Western blotting. Then, Tr.P32 was purified by Ni-NTA affinity chromatography under denaturing conditions and used to develop a capripoxvirus-specific ELISA. Checkerboard titration and receiver-operating characteristic (ROC) analysis were used to optimize the ELISA system and determine diagnostic specificity and sensitivity, respectively. The diagnostic potential of the developed ELISA was evaluated using positive and negative control sera collected from goat, sheep, and cattle. Results showed that the expression level of full-length P32 recombinant protein was negligible, while Tr.P32, a ~ 31 kDa recombinant protein, was expressed up to 0.270-0.300 mg/200 mL of culture media. The results of checkerboard titration revealed that 675 ng/well of Tr.P32 antigen and 1:10 dilution of control sera (anti GTPV HIS and healthy goat sera) caused maximum difference in absorbance between positive and negative goat sera. The recombinant Tr.P32 showed good reactions with antibodies against GTP virus (GTPV), SPP virus (SPPV), and LSD virus (LSDV), whereas no cross-reactions with anti-Orf virus antibodies were detected. By comparing with the neutralization index (NI), cut off, diagnostic sensitivity and specificity of the developed indirect-ELISA were estimated, 0.397, 94% and 96.6%, respectively. These findings indicate that the ELISA system based on Tr.P32 protein could potentially be used in sero-surveillance of all capripoxviruses; however, further investigations are required.

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Sheep and goat pox (SGPX) are systemic transmissible viral diseases characterized mainly by skin and internal lesions. This work aimed to provide the first molecular detection of sheep pox virus (SPV) in Iraq. Sheep pox virus (SPV) is a widely spread skin viral disease in Iraqi sheep and is associated with the formation of benign and malignant lesions in adult and young animals, resulting in notable economic losses in sheep and goat. In the current study, 24 sheep pox samples were collected from sheep from central Iraq. These samples were analyzed by polymerase chain reaction, and sequence analysis. Sheep pox virus DNA was detected in 18 of the samples (75%) in Iraqi sheep as the main causative agent for the disease. A partial sequence for the SGP P32 gene was deposited in Gene Bank. Phylogenetic analysis confirmed the presence of SPV-Iraqi isolate and showed that the origin of infection may be imported from India, china and USA sheep. This study presents the first report of SPV-infection in the Iraqi sheep and gives to spreading the knowledge of the origin of this disease. The results of this study will assist in the development of appropriate prophylaxis procedures and therapeutic measures.

H^t of occurrence of sheep and goat pox and peste des petits rum^nts requires development of new vaccines and improvement of available associated ones. Their administration allows for the reduction of vaccination time frames and cost. The study results demonstrated tat associated as well as monovalent virus vaccines against sheep pox, goat pox and peste des petits rum^nts have good antigenic properties. The vaccination induced high level of virus neutralizing antibodies against each of the above listed diseases and ensured formation of strong immunity tat is supported by the results of tallenge of sheep with virulent sheep pox virus (“Afghanistan” strain). In 14 and 21 days in blood of sheep immunized with the virus vaccine against sheep pox, goat pox and peste des petits rum^nts titres of antibodies against sheep pox virus ((4.17±0.12)-(4.84±0.31)) and PPR ((5.92±0.23)-(6.08±0.23)) were higher as compared to goat pox antibody titres ((1.99±0.89)-(3.53±0.58) log2). Herewith, antibody titres in blood sera of animals vaccinated with monovalent vaccine against sheep pox were higher against homologous virus ((3.62±0.12)- (4.08±0.31)) as compared to goat pox ((2.50±0.50)-(2.25±0.25) log2). The data demonstrate tat the associated vaccines may be used for preventive vaccination against sheep pox, goat pox and peste des petits rum^nts.

Antibodies were shown in blood of lambs vaccinated once with anti-sheep pox virus vaccine based "ARRIAH" strain at the age of one, two and three months and their titres were age-dependant. It was demonstrated that the sheep were resistant to the disease if their mean postvaccinal virus neutralizing antibody titres were within (1.75 ± 0.16)-(2.34 ± 0.24) log2. The regression models characterizing relationship between humoral factor levels and protective function of anti-sheep pox virus immunity in sheep were constructed. It was determined that 78% and 91% of animals in the flock should be seropositive with mean antibody titres of 1.43 log2 and 1.97 log2 to ensure protection of 80% and 95% of immunized animals, respectively.

Aim: The study was undertaken to assess the prevalence of antibodies to Capripoxviruses among small ruminants of Odisha, India. Materials and Methods: A total of 500 random serum samples collected from 214 sheep and 286 goats across 10 agro-climatic zones of Odisha, were screened using whole virus antigen-based indirect ELISA for antibodies against Capripoxviruses. Results were analyzed by suitable statistical methods. Results: Screening of 500 serum samples showed seropositivity of 8.88% and 31.47% in sheep and goats, respectively, for Capripoxviruses. The prevalence rate according to agro-climatic zone ranged from 0% (North Eastern coastal plain zone) to 48.57% (North central plateau zone) for goat pox, and 0% (Western undulating zone and North central plateau) to 22.22% (South Eastern ghat zone) for sheep pox. The difference in prevalence rates among the various agro-climatic zones was statistically significant (p

In this study, the ability of the combined vaccine against peste des petits ruminants (PPR) (Nigeria strain 75/1) and sheep pox (SPP) (NISKhI strain) to form a protective immune response for 12 months in Kazakh breed fine-fleeced sheep aged 6–12 months was demonstrated. The duration of the protective immunity of immunized sheep from PPR and from SPP was evaluated using a serum neutralization test (SNT), followed by testing of the resistance of vaccinated sheep to infection with the field strain Kentau-7 of the PPRV and the virulent strain A of the SPPV. The PPR antibody response was additionally measured by c-ELISA. A single immunization of sheep with a combined vaccine in a volume of 2.0 mL, containing the PPR and SPP vaccine viruses in the titers of 103.0 TCID50/mL, provided reliable protection of animals from two infections simultaneously for 12 months (observation period). At the same time, in sheep immunized with the combined vaccine, antibodies of PPRV persisted for up to 12 months, with slight fluctuations. The combined vaccine induced 100% clinical protection against the field strain of PPRV and the virulent strain of SPPV in immunized sheep for up to 12 months, while unvaccinated animals became ill with the manifestation of clinical signs specific to PPRV and SPPV.

Aim: To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt. Materials and Methods: A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened. Results: Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively. Conclusion: PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection.

In this review the authors analyzed the sheep pox disease, which occurs outbreaks all over the world particularly in Asia and Africa causing substantial losses in trade of animal and animal products. They categorized the sheep pox disease is one of the prioritized groups of diseases against which the World Organization for Animal Health is fighting. Data concerning a sheep poxes’ history, epidemiology, epizootiology, mortality and economic impact, clinical and pathological signs, features of capripoxvirus that forms the disease are given. Diagnosis treatment and vaccine have been investigated as well. The main conclusion is done according which the designing of new vaccine generation against the sheep pox disease could be as an alternative approach against sheep pox.

• SPPV is endemic in Algeria despite the implementation of a control program. • SPPV outbreaks occurred every years across all Algeria region. • Our study revealed the occurrence of an average of 449 outbreaks per year. • The neutralization index range between 0.73 and 1.22. Sheep pox is a disease of veterinary concern to small ruminant producers and veterinary diagnosticians, because of the associated tangible economic losses. The epidemiological analysis of sheep pox, among vaccinated sheep flock in Algeria from 2007 to 2016, showed that the disease outbreaks occurred every year and across all Algeria region with an average of 44.9 outbreaks per year, these outbreaks correlate with the region climate, the flocks' density and the transhumance practices. The one-year post vaccination antibody kinetics evaluation study of the commercially used vaccine in Algeria demonstrated a mild humoral response, the neutralization index range between 0.73 and 1.22. Therefore, the present study recommends a challenge study, using a virulent local strain, to evaluate the vaccine efficacy. Furthermore, quality control approach for the vaccine production processes is required. [ABSTRACT FROM AUTHOR]

Lumpy skin disease (LSD) is an emerging infectious disease of cattle. Since 2012, it has been seen throughout the Middle East region. The aim of this study was to compare the humoral response of three different dosages of the RM65 sheep pox (SPP) vaccine to assess the use of ten times sheep dose of the RM65 vaccine against lumpy skin disease, and to explore the possible causes of, and characterize the side effects caused by the RM65 vaccine. A blinded randomized collected study comprised 57 clinically normal, Holstein Friesian cattle which were randomly assigned into three experimental groups of 17 cattle according to the vaccine dose used (one, five and ten times the dose used for sheep in the field, and a control group of six cattle that did not receive the vaccine. Experimental animals were monitored closely for the development of any abnormality or side effects. Serum samples were collected for 6 weeks and were tested using serum neutralization assay. Decrease in total milk production was observed a week after vaccination and by the fifth week of the experiment, it had returned to prevaccination levels. Clinical side effects were seen in five animals that belong only to the group that received ten times of the SPP vaccine dose. Observed side effects included fever, decreased feed intake and milk production, as well as skin lesions. Skin nodules appeared between 7 and 17 days postvaccination, and remained for 11–17 days. Systemic reactions were likely to be associated with higher dosage and all affected cattle recovered uneventfully. Animals that received the highest dose (ten times the sheep dose) showed the best humoral response. The actual efficacy of the different concentration of the SPP RM65 should be evaluated based on a challenge experiment in a controlled environment. [ABSTRACT FROM AUTHOR]

Sheep and goat pox (SGPX) are systemic transmissible viral diseases characterized mainly by skin and internal lesions. This work aimed to provide the first molecular detection of sheep pox virus (SPV) in Iraq. Sheep pox virus (SPV) is a widely spread skin viral disease in Iraqi sheep and is associated with the formation of benign and malignant lesions in adult and young animals, resulting in notable economic losses in sheep and goat. In the current study, 24 sheep pox samples were collected from sheep from central Iraq. These samples were analyzed by polymerase chain reaction, and sequence analysis. Sheep pox virus DNA was detected in 18 of the samples (75%) in Iraqi sheep as the main causative agent for the disease. A partial sequence for the SGP P32 gene was deposited in Gene Bank. Phylogenetic analysis confirmed the presence of SPV-Iraqi isolate and showed that the origin of infection may be imported from India, china and USA sheep. This study presents the first report of SPV-infection in the Iraqi sheep and gives to spreading the knowledge of the origin of this disease. The results of this study will assist in the development of appropriate prophylaxis procedures and therapeutic measures. [ABSTRACT FROM AUTHOR]

Background and Objectives: Sheeppox virus causes systemic disease in sheep that is often associated with high morbidity and mortality. Protection against sheep pox is mainly based on medical prophylaxis, vaccination being the only way. In Morocco, and up to now, there is no available information about local challenge strain to use for controlling the efficiency of vaccines produced against sheep pox. Hence, the objective of the present study was to evaluate and compare the pathogenicity of seven Sheeppox virus (SPVs) isolates from 1993-1995 in Morocco. Materials and Methods: These seven SPV isolates have undergone various tests to evaluate their pathogenicity: Passages and titration on cell culture, Experimental inoculation on sheep, Virus-neutralization, In vivo titration and viral re-isolation by real-time PCR assay. Results: All infected lambs showed severe clinical signs, while most of them have been reproduced on 5 dpi and persisted until 21 dpi. The lambs infected by Oj1P4, Oj2P4 and BerP5 appeared lethargic, reluctant to move compared to those infected by other isolates. The results also revealed that all isolates were able to induce serological response. Virus isolation from infected organs and blood and amplification of the viral DNA by real-time PCR proved the presence of the virus in tissues and blood of infected lambs. These Moroccan SPVs demonstrated that the three isolates Oj1P4, Oj2P4 and BerP5 have a high pathogenicity; especially the BerP5 isolate which has an important infectious titer. Conclusion: These results demonstrate that the Berkane isolate is the most pathogenic of the tested isolates and it can be an excellent challenge strain for the control of the efficiency of vaccines against sheep pox produced in Morocco.

Abstract In this paper, we report on the development of a real time high-resolution melting (HRM) PCR assay for detection and differentiation among sheep pox virus (SPPV), goat pox virus (GTPV), field isolates and vaccine strains of lumpy skin disease virus (LSDV) based on high-resolution melting curve analysis of their target PCR amplicons. A 111 bp region of LSDV010 ORF, which harbors unique genetic differences for each of these viral species, was selected as the PCR target in this study. During the validation of this assay using DNA from clinical isolates originated from naturally infected animals from the different geographic locations and reference strains, the obtained PCR amplicons demonstrated that the melting temperature picks were specific for each tested viral species, i.e., 74.56 ± 0.04 °C for field LSDV, 74.95 ± 0.08 °C for vaccine LSDV, 74.24 ± 0.06 °C for SPPV and 73.61 ± 0.04 °C for GTPV. The assessment of the assay sensitivity utilizing a LSDV field strain as a PCR template revealed the assay detection limit as low as 0.1 TCD50 lg/ml. Overall, this assay based on Rotor-Gene Q (QIAGEN) platform was shown to be reproducible across replicates and operators and can be recommended as an additional diagnostic tool to the currently available molecular assays for detection and differentiation of the genus Capripoxvirus species, including the differentiation of vaccine strains of LSDV from field isolates. The assay can be used for detection of these viruses in animal- and insect-derived field specimens. Highlights • A high resolution melting PCR assay for the detection and discrimination among capripoxviruses. • The assay was sensitive and reproducible across multiple runs and was evaluated on a panel of reference and field samples. • This tool is an alternative means for routine surveillance of capripoxviruses. [ABSTRACT FROM AUTHOR]

Bluetongue (BT), is one of the OIE-listed major diseases of ruminants. Following the official report of BT virus serotype 3 (BTV-3) in a sheep in Cap Bon (Tunisia), blood and serum samples of ruminants were collected from some areas of Tunisia to further investigate the presence of this virus in the country. A quantitative real time RT-PCR has been first developed for the detection and quantitation of BTV-3 RNA from field specimens. Out of 62 collected blood samples, 23 were shown to be positive for BTV-3 RNA. Isolation on cell cultures was also possible from six samples. Genome sequencing revealed the circulation of two unrelated western strains of BTV-3, one circulating in Cap Bon and neighboring areas, and the other circulating nearby the border with Libya. The presence of a putative novel BTV serotype (BTV-Y TUN2017) in sheep introduced from Libya to Tunisia, genomically related to the BTV strain contaminating a commercially-available sheep pox vaccine and to BTV-26, has been also demonstrated. This finding highlights the pressing need for a prompt production and release of a novel inactivated BTV-3 vaccine to be used in case of emergence or proactively in the areas of Southern Europe at major risk of BTV introduction. The assessment of a novel vaccine will certainly exalt the role and importance of surveillance activities and collaboration with Northern African countries. [ABSTRACT FROM AUTHOR]

Summary: Sheep pox, a well‐known endemic capripox infection, has significant impacts on small ruminant populations in Tunisia. It is responsible for high economic losses throughout North Africa due to its enzootic nature and to the active animal transhumance existing in some governorates in Tunisia. The aim of this review was to analyse data gathered on annual vaccination campaigns designed to control its spread by reducing the level of endemicity and to describe diagnostic and management tools adapted to the Tunisian situation. Seasonal, temporal and spatial distributions of sheep pox outbreaks, as well as related clinical features, were found. It was concluded from this review that establishing strong herd immunization through individual animal immunization, creating adequate infrastructure, increasing awareness among breeders, setting up a field‐based surveillance network and improving routine diagnostic methods need to be the major components of a programme to eradicate the disease. It was also felt that cost‐benefit analyses of the surveillance and control strategies used would help in controlling its persistence. [ABSTRACT FROM AUTHOR]

Aim: The study was undertaken to assess the prevalence of antibodies to Capripoxviruses among small ruminants of Odisha, India. Materials and Methods: A total of 500 random serum samples collected from 214 sheep and 286 goats across 10 agro-climatic zones of Odisha, were screened using whole virus antigen-based indirect ELISA for antibodies against Capripoxviruses. Results were analyzed by suitable statistical methods. Results: Screening of 500 serum samples showed seropositivity of 8.88% and 31.47% in sheep and goats, respectively, for Capripoxviruses. The prevalence rate according to agro-climatic zone ranged from 0% (North Eastern coastal plain zone) to 48.57% (North central plateau zone) for goat pox, and 0% (Western undulating zone and North central plateau) to 22.22% (South Eastern ghat zone) for sheep pox. The difference in prevalence rates among the various agro-climatic zones was statistically significant (p<0.05) for goats, but not for sheep. Antibody prevalence rates among various districts were recorded to be the highest in Jagatsinghpur (30%) for sheep pox and Dhenkanal (80%) for goat pox. Conclusion: The study revealed serological evidence of Capripoxvirus infection in sheep and goat populations in the study area, in the absence of vaccination. Systematic investigation, monitoring, and reporting of outbreaks are necessary to devise control strategies. [ABSTRACT FROM AUTHOR]

Background:Sheep pox and goat pox are contagious viral diseases of sheep and goats, respectively. The diseases result in substantial economic losses due to decreased milk and meat production, damage to hides and wool, and possible trade restriction. A study was undertaken in Amhara region of Ethiopia. A cross-sectional study design was used to estimate the sero-prevalence and identify associated risk factors, while retrospective study design was used to assess the temporal and spatial distribution of the disease. A total of 672 serum samples were collected from 30 Kebeles and tested using virus neutralization test. Results: From a total of 672 sera tested, 104 (15.5%) were positive for sheep and goat pox virus antibody; from which 56 (17%) were sheep and 48 (14%) were goats. The diseases were prevalent in all study zones, the highest sero-prevalence was observed in South Gondar (20.9%) and the lowest in North Gondar and West Gojjam zones (11.9% each). From the potential risk factors considered (species, sex, age, agro-ecology and location); only sex and age were significantly associated (p < 0.05) with the diseases in multivariable logistic regression. Female and young animals were at higher risk than their counterparts. From January 2010 to December 2014, a total of 366 outbreaks, 12,822 cases and 1480 deaths due to SP and 182 outbreaks, 10,066 cases and 997 deaths due to GP were recorded in Amhara National Regional State. Conclusion: Both the serological and the outbreak data revealed that sheep and goat pox is one of the most prevalent and widespread diseases of sheep and goats in the study area. Hence, annual mass vaccination program must be implemented for economic and viable control of sheep and goat pox diseases in the Amhara region in particular and at a national level in general. [ABSTRACT FROM AUTHOR]

Background and Objectives: Sheeppox virus causes systemic disease in sheep that is often associated with high morbidity and mortality. Protection against sheep pox is mainly based on medical prophylaxis, vaccination being the only way. In Morocco, and up to now, there is no available information about local challenge strain to use for controlling the efficiency of vaccines produced against sheep pox. Hence, the objective of the present study was to evaluate and compare the pathogenicity of seven Sheeppox virus (SPVs) isolates from 1993-1995 in Morocco. Materials and Methods: These seven SPV isolates have undergone various tests to evaluate their pathogenicity: Passages and titration on cell culture, Experimental inoculation on sheep, Virus-neutralization, In vivo titration and viral re-isolation by real-time PCR assay. Results: All infected lambs showed severe clinical signs, while most of them have been reproduced on 5 dpi and persisted until 21 dpi. The lambs infected by Oj1P4, Oj2P4 and BerP5 appeared lethargic, reluctant to move compared to those infected by other isolates. The results also revealed that all isolates were able to induce serological response. Virus isolation from infected organs and blood and amplification of the viral DNA by real-time PCR proved the presence of the virus in tissues and blood of infected lambs. These Moroccan SPVs demonstrated that the three isolates Oj1P4, Oj2P4 and BerP5 have a high pathogenicity; especially the BerP5 isolate which has an important infectious titer. Conclusion: These results demonstrate that the Berkane isolate is the most pathogenic of the tested isolates and it can be an excellent challenge strain for the control of the efficiency of vaccines against sheep pox produced in Morocco. [ABSTRACT FROM AUTHOR]

Background: Clinically, sheeppox and goatpox have the same symptoms and cannot be distinguished serologically. A cheaper and easy method for differential diagnosis is important in control of this disease in endemic region. Methods: A duplex PCR assay was developed for the specific differential detection of Goatpox virus (GTPV) and Sheeppox virus (SPPV), using two sets of primers based on viral E10R gene and RPO132 gene. Results: Nucleic acid electrophoresis results showed that SPPV-positive samples appear two bands, and GTPVpositive samples only one stripe. There were no cross-reactions with nucleic acids extracted from other pathogens including foot-and-mouth disease virus, Orf virus. The duplex PCR assay developed can specially detect SPPV or GTPV present in samples (n = 135) collected from suspected cases of Capripox. Conclusions: The duplex PCR assay developed is a specific and sensitive method for the differential diagnosis of GTPV and SPPV infection, with the potential to be standardized as a detection method for Capripox in endemic areas. [ABSTRACT FROM AUTHOR]

The article discuses various developments associated with emergency animal disease management. It is noted that sheep pox and goat pox are highly contagious diseases that affect small ruminants, whose clinical signs include fever, papules and pustules on sparsely woolled or haired skin, hypersalivation and ocular and nasal discharge. It is noted that differential diagnoses include contagious pustular dermatitis, bluetongue, mycotic dermatitis, ectoparasites and photosensitisation.

Sheep pox (SP) still represents a major threat to animal wealth. Vaccination is the only reliable remedy for such highly infectious and contagious diseases. Chitosan nanoparticles (CNP) could be successfully added to live as well as inactivated vaccines to enhance their immune responses as it exhibits a good delivery system. In this study, CNP was added to maintenance essential media (MEM) with ratios of 3, 15 and 30 μg/ml. Such preparations were overlayed on confluent vero cell sheets for 7 days. CNP augmented MEM was proved to be unharmful for VERO cells. The prementioned concentrations of CNPMEM were mounted the infected sheets of cells that had been inoculated with the Romanian vaccinal strain of sheep pox virus (SPV). Appearance of the cytopathic effect as well as SPV infectivity titers were slightly improved particularly with CNP-MEM that was augmented with 15μg/ml. The former infected culture was used to prepare pilot batch of CNP supplemented sheep pox vaccine. The immune response of the pilot batch was assessed comparing with the conventionally produced SP vaccine by inoculating them into separate groups of susceptible sheep. There was slight increase in the serum neutralizing antibody titers in the group inoculated with the CNP augmented vaccine over the traditional one. It has been deduced that CNP could be used in sheep pox vaccine preparation to enhance the vaccine quality. [ABSTRACT FROM AUTHOR]

Capripoxviruses (CaPVs), consisting of Sheeppox virus (SPV), Goatpox virus (GPV), and Lumpy skin disease virus (LSDV) species, cause economically significant diseases in sheep, goats, and cattle, respectively. Quantitative real-time polymerase chain reaction (qPCR) assays are routinely used for rapid detection of CaPVs in surveillance and outbreak management programs. We further modified and optimized 2 previously published CaPV qPCR assays, referred to as the Balinsky and Bowden assays, by changing commercial PCR reagents used in the tests. The modified assays displayed 100% analytical specificity and showed no apparent changes in analytical sensitivities for detection of CaPVs compared with the original assays. Diagnostic sensitivities, assessed using 50 clinical reference samples from experimentally infected sheep, goats, and cattle, improved from 82% to 92% for the modified Balinsky assay and from 58% to 82% for the modified Bowden assay. The modified qPCR assays were multiplexed for detection of beta-actin as an indicator for potential false-negative results. The multiplex modified qPCR assays exhibited the same diagnostic sensitivities as the singleplex assays suggesting their utility in the detection of CaPVs. [ABSTRACT FROM AUTHOR]

The present study was aimed to asses clinical, histopathology, PCR and phylogenetic analysis of sheep pox virus (SPPV) in indigenous sheep. A sheep farm at Al Hassa District was struck by the sheep pox virus. The clinical signs were a sudden onset of fever, which peaks at 40-42°C, with discharges from the nose and eyes, and excessive salivation. The animal loses its appetite and is reluctant to move. Mortality was mostly seen in lambs. At necropsy, the skin lesions were erythematous macules and papules, measured 2 to 4 mm in diameter which may become larger and nodular. These lesions were frequently seen in face, back and tail. Multiple firm white nodules disseminated throughout the lobes of the lungs, particularly in lambs. Specific histopathologic features like epidermal hyperplasia with ballooning degeneration and the presence of intracytoplasmic eosinophilic inclusion bodies associated with hyperplasia of pneumocyte type II were seen in the affected animals. The collected samples were screened for the presence of SPPV DNAs using KS- 1.5/KS-1.6 and InS-1.1/Ins-1.1/based multiplex PCR. The p32 gene of selected two positive samples was sequenced and aligned with different SPPV, GTPV and LSDV available in GenBank. The phylogenetic analysis revealed that sheep pox virus strain; SPPV/Al-Hassa/2014/Saudi Arabia (accession number, KP342531) clustered on SPPV clad with SPPVs from India and China. [ABSTRACT FROM AUTHOR]

Sheep pox is endemic in most parts of Northern Africa and has the potential to cause severe economic problems. Live attenuated vaccines are used in Morocco, and in many other countries, to control the disease. Sheep pox virus ( SPPV) re-appeared in 2010 causing a nodular clinical form previously not observed in Morocco. The severe clinical signs observed during the course of this outbreak and initial reports citing similarity in nucleotide sequence between the Moroccan vaccine strain and field isolates warranted a more in depth analysis of this epizootic. In this study, sequence analysis showed that isolates obtained from four provinces of eastern Morocco were identical, demonstrating that a single SPPV strain was responsible for the 2010 epizootic. In addition, the genome fragments sequenced and phylogenetic analyses undertaken as part of this study showed significant differences between field isolates and the Moroccan vaccine strain. New PCR methods were developed to differentiate between wild-type isolates and vaccine strains of SPPV. Using these methods, no trace of wild-type SPPV was found in the vaccine and no evidence was found to suggest that the vaccine strain was causing clinical disease. [ABSTRACT FROM AUTHOR]

Aim: To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt. Materials and Methods: A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened. Results: Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively. Conclusion: PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection. [ABSTRACT FROM AUTHOR]

The objective of this immunohistochemical study was to examine the expression of caspase-3,-5,-7, and -9 in sheep pox lesions in 15 naturally infected crossbred merino lambs. Sheep pox was diagnosed based on typical gross and histopathological lesions in the skin and visceral organs that were evaluated pathologically and immunohistochemically. Macroscopically, lesions were observed in the skin and visceral organs. Characteristic sheep pox inclusion bodies and cellules claveleuses (sheep pox cells) were present in all cases. After histopathological examinations, tissue sections were immunostained for caspase-3,-5,-7, and -9. While strong positive reactions were observed in normal tissue, only weak expression was noted in poxvirus lesions, demonstrating marked caspase inhibition in naturally infected lambs. This finding suggests a role for caspases in sheep poxvirus infection and its pathogenesis. [ABSTRACT FROM AUTHOR]

Bluetongue (BT), peste des petits ruminants (PPR) and sheep pox are the most economically important viral diseases of sheep in India. Serum samples obtained from sheep in five northern states of the country were screened for antibody against these agents to explore the extent of spread of these infections. A total of 516 serum samples were screened for the presence of antibodies against BT and PPR viruses. Of these, 155 samples were also tested for antibodies against sheep pox virus. BT antibodies were found in 293 (56.8%) animals, PPR virus antibodies in 215 (41.7%) and sheep pox virus antibodies in 106 (68.3%). Of the serum samples tested, 25.2% were positive for antibodies against all three viruses. These findings clearly demonstrated not only the enzootic nature of disease, but also the co-existence of antibodies to more than one of these viruses which would indicate that concurrent infections were common. Therefore, control measures should focus in combating all three diseases simultaneously by exploring the possibility of a trivalent vaccine or the use of multiple genes expressing vectored vaccine.

Background: Sheeppox (SPP) is one of the priorities, high-impact animal diseases in many developing countries, where live attenuated vaccines are routinely used against sheeppox virus (SPPV). In an event of an SPP outbreak, historically disease-free countries would hesitate to use of live vaccines against SPPV due to the safety and trade reasons. Currently no killed SPPV vaccines are commercially available. In this study, we developed an inactivated Romanian SPPVvaccine and assessed its efficacy and potency in comparison with a live attenuated Romanian SPPV vaccine. Four naïve sheep were vaccinated once with the Romanian SPPV live attenuated vaccine and16 sheep were vaccinated twice with the inactivated vaccine. All sheep in the live vaccine group were included in the challenge trial, which was conducted using a highly virulent Moroccan SPPV field strain. Eight sheep of the inactivated vaccine group were challenged and the remaining sheep were monitored for seroconversion. Experimental animals were closely monitored for the appearance of clinical signs, body temperature and inflammation at the injection site. Two naïve sheep were used as unvaccinated controls. Results: The inactivated Romanian SPPV vaccine was found to be safe and confer a good protection, similar to the live vaccine. Specific antibodies appeared from seven days post vaccination and remained up to nine months. Conclusion: This study showed that the developed inactivated Romanian SPPV vaccine has a potential to replace attenuated vaccine to control and prevent sheep pox in disease-free or endemic countries. [ABSTRACT FROM AUTHOR]

In this report we describe the detection and identification of Bluetongue virus (BTV) contaminations in commercial vaccines. BTV RNA was detected in vaccine batches of Lumpy skin disease (LSD) and Sheep pox (SP) using quantitative PCR (qPCR) for VP1 and NS3 genes. Both batches were positive for VP1 and NS3 in qPCR. The LSD vaccine-derived sample was positive for VP1 and VP2 in conventional PCR. The SP vaccine-derived sample was examined by amplification of VP1, VP4, VP6, VP7, NS2 and NS3 gene segments in conventional PCR. The SP vaccine-derived sample was further propagated in embryonated chicken eggs (ECE) and Vero cells. Preliminary sequence analysis showed that the LSD vaccine-derived sequence was 98–99% similar to BTV9. Analysis of the six genomic segments from the SP vaccine-derived isolate showed the highest similarity to BTV26 (66.3–97.8%). These findings are particularly important due to the effect of BTV on cattle and sheep, for which the vaccines are intended. They also demonstrate the necessity of rigorous vaccine inspection and strict vaccine production control. [ABSTRACT FROM AUTHOR]

A Loop-mediated isothermal amplification (LAMP) assay targeting the highly conserved DNA polymerase gene of capripox virus genome was developed and evaluated for rapid detection of sheep pox and goat pox viruses. The optimized LAMP assay is found specific and sensitive for amplification of target DNA with a diagnostic sensitivity and specificity of 96.6% and 100% respectively compared to quantitative PCR. The detection rate of LAMP, PCR and Q-PCR assays is found to be 81.5%, 67% and 83% respectively. This LAMP assay has the potential for rapid clinical diagnosis and surveillance of sheep pox and goat pox in field diagnostic laboratories. [ABSTRACT FROM AUTHOR]