Sheep Pox Research Articles

Prevalence and PCR based molecular characterization of goat pox virus from field outbreaks of Multan and Bahawalnagar, Pakistan

This study was designed to check the prevalence and PCR-based molecular characterization of goat pox virus (GTPV) in the Multan and Bahwalnagar regions of Punjab, Pakistan. Capripox virus (CPPV) is the cause of goat pox (GTP) and sheep pox (SPP) disease, it highly affects the morbidity and mortality rate of goats and sheep. In this study, the 80 tissues and blood samples of goats were collected on age basis from the goat farms, slaughter houses, tanneries and domestic animals. The epidemiological data was also collected. The collected samples were processed for DNA extraction. We characterized the goat pox virus (GTPV) with specific primers of P32 gene by PCR. Then each amplified product was analyzed by agarose gel electrophoresis visualized by UV fluorescence light. This study showed that Infants of goats (2-10 months) in Multan showed 25% while adult goat in Multan showed 14.2 % positive results. In Bahawlnagar, the affected infants of goats (2-10 months) found were 31.25% while adult infected goats were 11.1%. Both primers were equally effective for the characterization of unknown samples. The most effected goats were adult female and infants.
 J. bio-sci. 28: 95-103, 2020

Poxvirus encoded eIF2α homolog, K3 family proteins, is a key determinant of poxvirus host species specificity

Protein kinase R plays a key role in innate antiviral immune responses of vertebrate animals. Most mammalian poxviruses encode two PKR antagonists, E3 (dsRNA binding) and K3 (eIF2α homolog) proteins. In this study, the role of K3 family proteins from poxviruses with distinct host tropisms in determining the virus host range was examined in a vaccinia E3L deletion mutant virus. It was found that K3 orthologs from the species-specific poxviruses (taterapox virus, sheeppox virus, myxoma virus, swinepox virus and yaba monkey tumor virus) restored the virus replication competency in cells derived from their natural hosts or related animal species. Further, it was found that the residues located in the helix insert region of the protein, K45 of vaccinia K3 and Y47 of the sheep poxvirus ortholog 011, are critical for the virus host species specificity. These observations demonstrate that poxvirus K3 proteins are major determinants of the virus host specificity.

Potential of Using Capripoxvirus Vectored Vaccines Against Arboviruses in Sheep, Goats, and Cattle.

The genus capripoxvirus consists of sheeppox virus, goatpox virus, and lumpy skin disease virus, which affect sheep, goats, and cattle, respectively. Together capripoxviruses cause significant economic losses to the sheep, goat, and cattle industry where these diseases are present. These diseases have spread into previously free bordering regions most recently demonstrated with the spread of lumpy skin disease virus into the Middle East, some Eastern European countries, and Russia. This recent spread has highlighted the transboundary nature of these diseases. To control lumpy skin disease virus, live attenuated viral vaccines are used in endemic countries as well as in response to an outbreak. For sheeppox and goatpox, live attenuated viral vaccines are used in endemic countries; these diseases can also be contained through slaughter of infected animals to stamp out the disease. The thermostability, narrow host range, and ability of capripoxviruses to express a wide variety of antigens make capripoxviruses ideal vectors. The ability to immunize animals against multiple diseases simultaneously increases vaccination efficiency by decreasing the number of vaccinations required. Additionally, the use of capripoxvirus vectored vaccines allows the possibility of differentiating infected from vaccinated animals. Arboviruses such as bluetongue virus and Rift Valley fever viruses are also responsible for significant economic losses in endemic countries. In the case of Rift Valley fever virus, vaccination is not routinely practiced unless there is an outbreak making vaccination not as effective, therefore, incorporating Rift Valley fever vaccination into routine capripoxvirus vaccination would be highly beneficial. This review will discuss the potential of using capripoxvirus as a vector expressing protective arboviral antigens.

Capripoxviruses: Exploring the genetic relatedness between field and vaccine strains from Egypt.

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.

Review on Alternative Vaccine of LSD

Lumpy skin disease is of significant economic impact for the cattle industry in Africa. Due to cross protection within the Capri poxvirus genus, sheep poxvirus (SPPV) vaccines have been widely used for cattle against lumpy skin disease virus (LSDV). In the Middle East and the Horn of Africa, these vaccines have been associated with incomplete protection and adverse reactions in cattle post vaccination. The different studies confirm that the real identity of the commonly used Kenyan sheep and goat pox vaccine virus (KSGP) O-240 is not SPPV but is actually LSDV. The low-level attenuation of this virus is likely to be not sufficient for safe use in cattle, causing clinical disease in vaccinated animals. In addition, Isiolo and Kedong goat pox strains, capable of infecting sheep, goats and cattle are identified for potential use as broad-spectrum vaccine candidates against all capripox diseases.

Pastoralist knowledge of sheep and goat disease and implications for peste des petits ruminants virus control in the Afar Region of Ethiopia

Pastoralist areas of Ethiopia are vulnerable to drought, causing livelihood loss and famine. One approach to increasing pastoralist resilience is the control of livestock disease, but there is limited information from pastoralist areas to inform control strategies. This study aimed to explore pastoralist concepts of small ruminant disease and implications for infectious disease surveillance and control in the pastoralist Afar Region.During 2013–14, qualitative and quantitative methods were applied in two villages of one district in the mid-west of the region. Semi-structured group interviews, incorporating participatory tools, explored pastoralist knowledge of small ruminant diseases and their impact. These were followed by multiple visits in different seasons to 70 households for semi-structured and informal interviews, observation of management practices, clinical examinations, and weekly questionnaires of mortality and morbidity. Thematic analysis was applied to interview transcripts and field notes, and descriptive statistical analysis to quantitative data.Afar concepts of disease causation, terminology and treatment were predominantly naturalistic, related to observable signs and physical causes, rather than personalistic factors (misfortune due to magical or spiritual agents). Disease occurrence was associated with malnutrition and adverse weather, and disease spread with contact between animals during grazing, watering and migration. Disease occurrence varied by season with most syndromes increasing in frequency during the dry season.Names for disease syndromes were related to the main clinical sign or body part affected; 70 terms were recorded for respiratory syndromes, diarrhoea, sheep and goat pox, lameness, skin diseases, ectoparasites, urinary and neurological syndromes and abortion. Some syndromes with pathognomonic signs could be linked to biomedical diagnoses but most were non-specific with several possible diagnoses. The syndromes causing greatest impact were diarrhoea and respiratory disease, due to mortality, reduced milk production, weight loss, abortion, weak offspring and reduced market value. Afar applied a range of traditional methods and modern medicines to prevent or treat disease, based on livestock keeper knowledge, advice of local specialists and occasionally advice from district veterinarians or animal health workers.In relation to surveillance for peste des petits ruminants (PPR), several terms were used for PPR-like syndromes, depending on the predominance of respiratory or diarrhoea signs. Therefore, whenever these terms are encountered during surveillance, the associated disease events should be fully investigated and samples collected for laboratory confirmation. The Afar naturalistic concepts of disease parallel biomedical concepts and provide a good foundation for communication between veterinarians and pastoralists in relation to PPR surveillance and control measures.

Genetic studies of terminal regions of vaccine and field isolates of capripoxviruses

Sheeppox and goatpox are two of the most important diseases associated with significant economic loss and impact on animal trade. In spite of the use of vaccines, outbreaks are being reported on several occasions. Therefore, deciphering the host specificity and virulence of sheeppox virus (SPPV) and goatpox virus (GTPV) is important in developing effective vaccines. It is opined that genes located in the terminal regions play a major role in determining host range and/or virulence. In the present study, nine isolates (6 GTPV and 3 SPPV; included both vaccine and virulent viruses) were genetically characterized by targeting 11 genes (7 host-range and 4 virulence genes) which are located in the terminal regions of capripoxviruses. In the genetic analyses, it was observed that there are several nucleotide and amino acid signatures which are specific for either SPPV or GTPV. However, surprisingly, none of the 11 genes could be able to differentiate the vaccine and field viruses of GTPV and SPPV. Our study indicates that the genes of the terminal regions may have a role in determining the host-specificity but the involvemet in determinatin of virulence/attenuation is not certain at least for the isolates used in the current study. Therefore, it is likely that some other genes located in terminal/central regions may also play a role in determination of virulence and pathogenesis which needs to be confirmed by whole-genome sequencing of several vaccine and virulent viruses.

OVERVIEW OF THE EPIDEMIC SITUATION ON CERTAIN INFECTIOUS ANIMAL DISEASES IN THE RUSSIAN FEDERATION IN 2018

The paper presents the results of analysis of epidemic situation on infectious animal diseases subject to compulsory OIE notification: FMD, African swine fever, classical swine fever, lumpy skin disease, sheep and goat pox, and highly pathogenic avian influenza. The paper presents data on the specified diseases situation in the Russian Federation, broken down by Subjects, and also provides a comparative description of the data for 2017 and 2018. Besides, attention is focused on some features of the epidemic process, depending on the season. For a long time the Russian Federation has been infected with several highly dangerous animal diseases, such as African and classical swine fever, FMD, lumpy skin disease, sheep and goat pox, and highly dangerous avian influenza. The specified above infections cause great damage to animal husbandry in the country which has a negative impact on its export potential. Analysis of the ASF epidemic situation demonstrates that the Russian Federation has been permanently infected and there is a trend for the geographic spread of the epidemic. Despite the fact that the number of ASF outbreaks newly detected in 2018 is almost half that of 2017, the situation remains tense. The forecast is still unfavorable. Lumpy skin disease has been reported in the Russian Federation for four years already. Herewith, from year to year the disease has been invading new territories. The HPAI epidemic situation is of great concern as in 2018 82 outbreaks were reported in poultry population which is 2.3 times more than in 2017. Besides, there is a trend for the epidemic extension. Thus, in 2018 avian influenza was reported in previously disease free Subjects of the Russian Federation. In 2018 the Russian Federation remained permanently infected with FMD, sheep and goat pox, and classical swine fever.

Comparative analysis of miRNAs in exosomes released by sheeppox virus-infected ovine testicular cells

Exosomes, secreted by various cells, are nanometer-scale vesicles with the functions in intercellular communication. To understand a role of exosomal miRNAs in the sheeppox virus infection, exosomes were isolated from sheeppox virus-infected sheep testicular cells 0 h, 24 h and 72 h post infection. The results of transmission electron microscopy and size distribution showed that all three exosome samples were spherical particles with negatively-stained membrane, ranging from 39 nm to 127 nm in diameter. A total of 106 known and 279 novel miRNAs were identified, and 78 known and 54 novel miRNAs were commonly detected in three exosome samples. Compared with the exosomes by the uninfected controls, a total of 34 known miRNAs were aberrantly expressed in the exosomes from infected cells. In agreement with the sequencing data, the expression of oar-miR-21 and oar-miR-10b was shown to be the highest in exosomes at 24 h after SPPV-infected, and the expression of oar-let-7f was the highest in exosomes at 72 h. Conversely, the expression of oar-let-7b and oar-miR-221 was significantly decreased 24 h and 72 h post infection compared with 0 h. The analysis results also revealed that differentially expressed miRNAs were mostly involved in an immune system process and stimulus response. These results provide rich data to further investigate a role of exosomal miRNAs in SPPV-host interactions.

Outbreak Investigation of Sheep pox in District Barkhan, Balochistan, Pakistan

During mid-January 2018, significant number of suspected sheep pox cases was reported to Director Animal Health from district Barkhan, Balochistan. An investigation team was deployed after unusual high mortality and complaint of skin lesions accompanied by lacrimal & nasal discharge. The aim of this investigation was to know the magnitude of the disease, identify risk factors and implement preventive measures so as to stop further spread of disease. We defined a sheep pox confirmed case as sheep with fever >40˚C and necrotic skin lesions along with one or more of the following: Papules, nodules anywhere on the body, Mucopurulent nasal discharge, hyperemia, oedema (generalized), swollen eyelids and enlarged prescapular lymph node. By using pre tested questionnaire rates and frequencies were calculated using Epi InfoTM 7 Software. A case control study was conducted in March 2018. During active case finding, 43 farms qualified as suspected for sheep pox infection with a total infected population of 4050 animals. Of these, infected adult sheep comprised 76% (3081), followed by 24% (969) lambs. The case fatality rate was 33% (1337/4050) with an overall attack rate of 55% (4050/7438). Majority 49% (21) responded that wool shearing within a flock was done, 42% (18) introduced new animals in a flock without prior quarantine but only 5% (2) were aware of isolation and 21% (9) vaccinate the animals. New animals with in flocks (OR: 3.7, CI 1.34 – 10, P value 0.008) and wool shearing (OR: 3.15, Cl 1.24 – 7.9, P value 0.01) were statistically associated with disease spread. Sheep pox outbreak occurred in the area with new animal arrival, low vaccine coverage and breach in biosecurity and sanitary measure. Standard biosecurity and sanitary prophylaxis measures were recommended to control the outbreak.

Extended sequencing of vaccine and wild-type capripoxvirus isolates provides insights into genes modulating virulence and host range.

The genus Capripoxvirus in the subfamily Chordopoxvirinae, family Poxviridae, comprises sheeppox virus (SPPV), goatpox virus (GTPV) and lumpy skin disease virus (LSDV), which cause the eponymous diseases across parts of Africa, the Middle East and Asia. These diseases cause significant economic losses and can have a devastating impact on the livelihoods and food security of small farm holders. So far, only live classically attenuated SPPV, GTPV and LSDV vaccines are commercially available and the history, safety and efficacy of many have not been well established. Here, we report 13 new capripoxvirus genome sequences, including the hairpin telomeres, from both pathogenic field isolates and vaccine strains. We have also updated the genome annotations to incorporate recent advances in our understanding of poxvirus biology. These new genomes and genes grouped phenetically with other previously sequenced capripoxvirus strains, and these new alignments collectively identified several recurring alterations in genes thought to modulate virulence and host range. In particular, some of the many large capripoxvirus ankyrin and kelch-like proteins are commonly mutated in vaccine strains, while the variola virus B22R-like gene homolog has also been disrupted in many vaccine isolates. Among these vaccine isolates, frameshift mutations are especially common and clearly present a risk of reversion to wild type in vaccines bearing these mutations. A consistent pattern of gene inactivation from LSDV to GTPV and then SPPV is also observed, much like the pattern of gene loss in orthopoxviruses, but, rather surprisingly, the overall genome size of ~150kbp remains relatively constant. These data provide new insights into the evolution of capripoxviruses and the determinants of pathogenicity and host range. They will find application in the development of new vaccines with better safety, efficacy and trade profiles.

TESTING SHEEP AND GOAT POX VIRUSES FOR THEIR REPRODUCTION IN PRIMARY AND SUBCULTURED CELLS

Results of tests of sheep and goat poxviruses for their reproduction in primary and subcultured cell cultures derived from lamb and goat kid kidneys and testicles are presented. Monolayer cultures were subcultured by 5 passages in plastic vials and infected with sheep and goat poxviruses. It was shown that production ARRIAH strain of sheep pox virus and ARRIAH 2003 strain of goat pox virus successfully propagated both in primary lamb and goat kid kidney and testicle cell cultures and lamb and goat kid kidney and testicle cell subcultures. Activity of sheep and goat poxviruses passaged 5 times was 5.5–6.0 lg TCID50/cm3. Taking into account that modern cell cultivation conditions allow primary trypsinized cell populations subcultivation up to 25–30th passage, subcultures together with continuous cell lines can be used for large-scale sheep and goat poxvirus production and research purposes.

Understanding the epidemiology of sheep-pox outbreaks among vaccinated Algerian sheep and post vaccination evaluation of the antibodies kinetics of the commercially used vaccine

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.

An HRM Assay to Differentiate Sheeppox Virus Vaccine Strains from Sheeppox Virus Field Isolates and other Capripoxvirus Species

Sheep poxvirus (SPPV), goat poxvirus (GTPV) and lumpy skin disease virus (LSDV) affect small ruminants and cattle causing sheeppox (SPP), goatpox (GTP) and lumpy skin disease (LSD) respectively. In endemic areas, vaccination with live attenuated vaccines derived from SPPV, GTPV or LSDV provides protection from SPP and GTP. As live poxviruses may cause adverse reactions in vaccinated animals, it is imperative to develop new diagnostic tools for the differentiation of SPPV field strains from attenuated vaccine strains. Within the capripoxvirus (CaPV) homolog of the variola virus B22R gene, we identified a unique region in SPPV vaccines with two deletions of 21 and 27 nucleotides and developed a High-Resolution Melting (HRM)-based assay. The HRM assay produces four distinct melting peaks, enabling the differentiation between SPPV vaccines, SPPV field isolates, GTPV and LSDV. This HRM assay is sensitive, specific, and provides a cost-effective means for the detection and classification of CaPVs and the differentiation of SPPV vaccines from SPPV field isolates.

Result of propagation of sheep and goat pox viral vaccine strains into vero cells

Background: Sheep and goat pox viral disease, which affects negatively to our country’s economy by prevalence and infection, has high mortality and morbidity rate. Although our country manufactures sheep and goat pox viral vaccine using lamb’s testicle tissue in the Biocombinat (Bio-factory), in winter, there is high number of diseased animals, lamb testicle is scarce, therefore there is a need to produce cell-culture based sheep and goat pox vaccine.
 Materials and methods: Russian VNIIZJ strain type 2 sheep pox vaccine and Chinese goat pox live vaccine (serial number 010030) antigens were used after 20 times of dilution and propogated into Vero and BHK-21 cell culture. By PCR the result was examined and sequenced by ABI3130xl sequencer machine and sequences were compared by MEGA7 program
 Results: 3 days after infection sheep pox and 4 days after infection goat pox were shown CPE (cytopathic effect or cytopathogenic effect) respectively, 6 days after the infection both of them has been shown full CPE. From these infected materials we isolated viral DNA and run PCR assay using Biocombinat’s strain as a positive control. PCR products were all equal, 289 bp long. When we compare these DNA sequences of sheep and goat pox viruses, they were 92% identical to complete genome of Indian sheep and goat pox virus.
 Conclusion: We can produce cell-culture based live vaccine and diagnostic tests for sheep and goat pox viral disease by re-propagating these strains into Vero.
 Хонь, ямааны цэцгийн вирусын вакцины омгийг vero эсэд дасгасан дүн
 Хураангуй: Хонь, ямааны цэцгийн вируст өвчин халдвар, тархалт өндөртэй бөгөөд өвчлөл, хорогдлынтүвшин өндөр байгаа нь манай улсын эдийн засагт сөргөөр нөлөөлж байна. Манай улс Биокомбинат (Био-үйлдвэр) үйлдвэрт хонины цэцэг өвчний эсрэг вакциныг хурганы төмсөгний анхдагч эдэд өсгөвөрлөх замаар амьд вакцин, ямааны цэцэг өвчний эсрэг вакциныг ямаанд халдвар хийж ам, хамрын орчинд үүссэн шархны эдийг химийн бодисоор идэвхгүйжүүлэх замаар тус тус үйлдвэрлэж байна. Ялангуяа хонины цэцэг өвчний вакциныг зөвхөн мал төллөх хаврын цагт үйлдвэрлэх боломжтой байдаг нь өвлийн цагт гарсан өвчнийг хянах аргагүйд хүрч байна. Иймээс цаг хугацаанаас хамааралгүй вакцин үйлдвэрлэхэд дамжмал эсийн технологи хэрэглэх зайлшгүй шаардлагатай байна. Оросын VNIIZJ омогашигласан 2 төрлийн хонины цэцгийн амьд вакцин, Хятадын ямааны цэцгийн амьд вакцин (серийн дугаар 010030) шингэлж, Vero эсийн өсгөвөрт халдаасан. Стандарт ПГУ-аар үр дүнг шалгаж, ABI3130xl sequencer машинаар үүсгэгчийн нуклеотидын дарааллыг тодорхойлсон ба MEGA7 програмаар дарааллуудыг харьцуулж удам зүйн мод байгуулсан. Эсэд халдвар хийснээс 3 хоногийн дараа хонины цэцэг, 4 хоногийн дараа ямааны цэцэг CPE (cytopathic effect) буюу эс эмгэгшүүлэх нөлөө тус тус үзүүлсэн бөгөөд 6 хоногийн дараа хоёулаа бүрэн CPE үзүүлсэн байна. Эдгээр вирус агуулсан эсийн тэжээлт орчноос бид вирусын ДНХ ялгаж Биокомбинат-ын хонины цэцгийн Перего омгийг эерэг хяналт болгон ашиглаж ПГУ-ын шинжилгээг явуулсан. ПГУ-ын бүтээгдэхүүн бүгд адилхан, 289 хос суурийн урттай байсан. Хонь, ямааны цэцгийн вирусын ДНХ-ийн нуклеотидын дарааллыг харьцуулж үзэхэд хониныцэцгийн вирусын бүтэн геномтой 92%, ямааны цэцгийн вирусын бүтэн геномтой 90% адилханбайв. Бид эдгээр өвчний үүсгэгчийг Vero эсэд халдвар хийх нөхцлийг тогтворжуулсан ньдамжмал эсийн өсгөвөрт суурилсан амьд вакцины түүхий эдийг ихээр бэлтгэх боломжтойболлоо.
 Түлхүүр үг: эсийн өсгөвөр, эсийн эмгэгшил, днх дараалал, амьд вакцин

Presence of bluetongue and epizootic hemorrhagic disease viruses in Egypt in 2016 and 2017

BTV and EHDV are closely-related orbiviruses that are transmitted between domestic and wild ruminants via the bites of hematophagous midges. Previous studies have reported seropositivity against BTV antibodies in sheep and goats in two Egyptian governorates (Beni Suef and Menoufia). However, no recent data are available on the BTV serotype(s) circulating in Egypt and the likely presence of EHDV has never been explored. This study investigated the presence of BTV and EHDV among cattle which had been found BTV-seropositive by ELISA method. These cattle living in proximity to sheep and goats previously found BTV-seropositive. These cattle displayed no clinical signs of BT but reproductive problems had been reported in herds. A total of 227 cattle blood samples were therefore collected in 2016 and 2017. Ninety-four of the 227 animals tested by a BTV ELISA were positive for BTV antibodies (41.4%). Of these 94 ELISA-positive cattle, only 83 EDTA-blood samples were available and therefore tested for BTV and EHDV genome detection by RT-PCR and sequencing.Of the cattle sampled in 2016, results revealed that two were RT-PCR-positive for BTV and seven for EHDV. Sequencing showed the presence of EHDV-1 and BTV-3 genome sequences. EHDV-1 S2 shared 99.5% homology with an EHDV-1 S2 from a strain isolated in 2016 in Israel. BTV-3 S2 and S8 sequences shared >99.8% nucleotide similarity with the BTV-3 Zarzis S2 and S8 sequences (Tunisian BTV, also detected in 2016). Of the 66 blood samples tested following their collection in 2017, they were all EHDV-negative by RT-qPCR while five were BTV- positive by RT-qPCR. However, attempts to identify the BTV serotype of these five samples were unsuccessful. Only part of BTV S8 was sequenced and it showed 79% nucleotide similarity with S8 of atypical BTV serotypes (particularly with BTV-26 and another BTV serotype strain isolated from a sheep pox vaccine). Overall, these findings demonstrate that both BTV and EHDV were circulating in Egypt in 2016 and 2017.

Trends and future prediction of livestock diseases outbreaks by periodic regression analysis

Livestock disease outbreaks become a burden to the animal husbandry farmers and cause great economic loss in India. Period regression analysis is used to find the periodic or cyclic character of livestock disease outbreaks in animals, as many other natural phenomena in environment is periodic or cyclic in nature. In present study, livestock disease outbreaks of anthrax (AX), black quarter (BQ), enterotoxaemia (ET), haemorrahgic septicemia (HS), bluetongue (BT), foot-and-mouth disease (FMD), peste des petits ruminants (PPR), sheep and goat pox (SGP), babesiosis (BA), fasciolosis (FA), theileriosis (TH) and trypanosomosis (TR) were analyzed using periodic regression to know the trend and future prediction of outbreaks. Time series data on disease outbreaks, month and year was collected from National Animal Disease Referral Expert System database for 2001–2016. The regression curves were prepared with baseline, observed outbreaks and upper bound curves for 12 livestock diseases. The analysis revealed decreasing trend for AX, BQ, ET, HS, FMD, PPR, SGP and a cyclical trend of peak occurrence for every 4–5 years was observed in BQ, PPR, SGP, FA and TR. However, TR showed increasing trend and BT, BA, FA, TH outbreaks were maintained at the same trend in the past and future also. Further, BQ in 2026, ET in 2020, HS in 2022, FMD in 2023, outbreak numbers may touch the zero point, if the preventive measures are continued for these diseases effectively. Thus, continuous and constant efforts are needed for prevention of livestock diseases outbreaks from all stakeholders, which will improve the economy of farmers in India.

RISK OF INTRODUCING HIGHLY DANGEROUS ANIMAL VESICULAR DISEASES INTO THE RUSSIAN FEDERATION

Animal vesicular diseases are rather widespread in the world. Transboundary vesicular diseases are mainly registered in African and Asian countries. They primarily include foot and mouth disease, sheep and goat pox, peste des petits ruminants, lumpy skin disease, swine vesicular disease, vesicular stomatitis, as well as relatively new Seneca Valley virus infection. The transboundary spread of vesicular diseases is mainly caused by legal and illegal movements of animals, animal products, feeds, live vaccines; migration of wild animals across the borders from neighboring countries and passive mechanical transmission of infection. The risk of vesicular disease agent introduction with legal import of live animals into the territory of the Russian Federation is minimal. Most likely, the causative agent can be introduced when livestock products are illegally imported from the following countries: China, Turkey, India, Mongolia, Iran, Kazakhstan, etc. Violations of foreign trade regulations pose a direct threat to the epidemic situation and economic security of Russia. There is a possibility of the introduction of vesicular agents with animal feed imports into the country since some viruses can potentially survive for a long time in some ingredients. Taking into account the fact that some neighboring countries are infected with vesicular diseases, the threat of infection introduction into Russia through migration of wild animals along the North Caucasian, Central Asian and Far Eastern routes still remains.

IMMUNOLOGICAL RESPONSE OF A NEW TRIVALENT CAPRIPOXVIRUS VACCINE IN PREGNANT EWES AND DOES

This study was conducted as a field trial of using a new candidate trivalent Capripoxvirus (CaPVs) vaccine in pregnant ewes and does to detect safety degree and level of immunity, in comparison with commercial Romanian sheep pox viral (RSPPV) vaccine. Monitoring the post-vaccinal reaction and measurement of cellular and humoral immunity by evaluating the lympho­cyte proliferation assay (LPA) and serum neutralization test (SNT), respectively, in addition to detect the acquired maternal immunity in new-born lambs and kids .The post-vaccine reaction in animals vaccinated with trivalent CaPVs vaccine was more noticeable than that vaccinated with RSPPV vaccine without significant difference (P-Value>0.05). In vitro, the evaluation of cel­lular and humoral immunity revealed that the animals vaccinated with trivalent vaccine can significantly induce lymphocyte proliferation response in addition to of high level of antibodies (Abs) in comparison to that vaccinated with RSPPV vaccine (P-Value <0.05) with peak titre of lymphocyte proliferation and Abs titre at 10th and 28th, respectively. Furthermore, the evaluation of maternal immunity in new-born lambs and kids from previously vaccinated dams with trivalent CaPVs and RSPPV vaccines by SNT was shown that the peak level of receiving immunity was appeared at 1st week after birth with increasing its level in lambs and kids of dams vaccinated with trivalent CaPVs vaccine than those of dams vaccinated with RSPPV vaccine with a significant difference in between (P-Value <0.05). The result of this study provide a good profile about a high immune response with a long level of maternal immunity provided with trivalent CaPVs vaccine in comparison to the RSPPV vaccine so toward about the preferable use of new vaccine to control infection with sheep and goat pox diseases. Trivalent CaPVs vaccine could be candidate to be used against lumpy skin disease (LSD) due to antigenic relationship between genus CaPVs strains.Key words: does; ewes; monovalent RSPPV; safety; trivalent CaPVs

A framework for reviewing livestock disease reporting systems in high-risk areas: assessing performance and perceptions towards foot and mouth disease reporting in the Thrace region of Greece, Bulgaria and Turkey.

Disease reporting is an essential frontline component of surveillance systems, particularly for detecting incursions of new and emerging diseases. It has the advantages of being comprehensive and continuous, with the potential to reduce the time of disease detection and the extent of consequent spread. A number of exotic diseases, including sheep and goat pox, lumpy skin disease, peste des petits ruminants and foot and mouth disease have historically entered into south-eastern Europe through the Thrace region, which extends across neighbouring areas of Greece, Bulgaria and Turkey. In this high-risk area, multiple factors can reduce the sensitivity of disease reporting across the diverse production systems and animal health services need robust and effective disease reporting systems. While describing a training exercise designed to provide animal health services of the three countries with the knowledge and skills for conducting comprehensive in-country assessments, we provide an initial evaluation of the sensitivity of foot and mouth disease reporting and identify gaps and constraints in the Thrace region. An expert elicitation approach was used to consult official veterinarians from central and local animal health authorities of the three countries, and scenario trees modelling was applied to analyse the collected data. The reported sensitivity of disease reporting often varied between the central and local veterinary authorities within the three countries. Awareness of clinical disease, of reporting procedures and of biosecurity measures affected the early stages of disease reporting, particularly in the production systems identified at lower reporting sensitivity such as small ruminant's herds, mixed bovine herds and backyard herds. Despite its limitations this training exercise provided an effective framework (a) to develop capacities of the veterinary services in the region and (b) to supply initial evidence for guiding further interventions targeting those sectors and stakeholders at lower reporting sensitivity to reduce risks of disease introduction.