Contagious Ecthyma Research Articles

Vacina experimental produzida em cultivo celular confere proteção parcial contra o ectima contagioso em ovinos

O ectima contagioso (também conhecido como orf), é uma doença debilitante de ovinos e caprinos causada pelo vírus do orf (ORFV). A vacinação tem sido usada com relativo sucesso no controle da doença. No entanto, as vacinas atuais contêm amostras virulentas do agente, são produzidas por escarificação cutânea de animais, e apresentam eficácia questionável. Assim, o presente trabalho teve como objetivo produzir e testar a eficácia de uma vacina experimental produzida em cultivo celular. A cepa IA-82 do ORFV foi submetida a 21 passagens em cultivo de células BHK-21 e usada para vacinar ovinos jovens (n=30), por escarificação cutânea na face interna da coxa. A vacinação produziu pústulas e crostas em 16 dos 30 ovinos vacinados, indicando imunização adequada. Noventa dias após a vacinação, ovinos vacinados (n=16) e controles (n=16) foram inoculados com uma cepa virulenta do ORFV (10(6,9)DICC50/mL) após escarificação na comissura labial. Todos os animais desenvolveram lesões típicas de ectima, incluindo hiperemia, vesículas, pústulas e crostas. No entanto, os animais vacinados desenvolveram lesões mais leves e passageiras do que os controles, e os escores clínicos foram estatisticamente diferentes (p<0,05) entre os dias 10 e 22 pós-desafio. Além disso, o tempo de duração da doença foi significativamente inferior (p<0,05) nos animais vacinados. Os animais vacinados também excretaram menor quantidade de vírus (p<0,05) e por um período significativamente mais curto do que os controles (13 dias versus 22 dias, p<0,001). Esses resultados demonstram a proteção parcial conferida pela vacina experimental e, dependendo da melhoria dos índices de imunização e proteção, são promissores no sentido da utilização de vacinas contra o ORFV produzidas em cultivo celular.

Evaluation of homologous and heterologous protection induced by a virulent field strain of orf virus and an orf vaccine in goats

To evaluate cross protection provided by administration of contagious ecthyma vaccines against strains of orf virus in goats. 126 Boer-Spanish crossbred goats (3 to 20 days old). 85 goats were vaccinated with a goat-derived contagious ecthyma vaccine. Of these, 41 were challenge exposed with the virus strain for the contagious ecthyma vaccine, 40 were challenge exposed with a more virulent field strain of orf virus, and 4 were lost to predation or died. Another 41 goats were vaccinated with a vaccine produced from a more virulent field strain of orf virus; of these, 18 were challenge exposed with the virus strain of the goat-derived contagious ecthyma vaccine, 18 were challenge exposed with the more virulent field strain of orf virus, and 5 were lost to predation or died. Vaccination with the goat-derived contagious ecthyma vaccine did not significantly reduce the number of goats with lesions or lesion severity caused by challenge exposure with the more virulent field strain of orf virus. Vaccination with the vaccine produced from the more virulent field strain of orf virus significantly reduced the number of goats with lesions attributable to challenge exposure with the virus strain of the goat-derived contagious ecthyma vaccine, but it failed to significantly reduce lesion severity. Vaccination did not result in cross protection for the 2 strains of orf virus. This may have been attributable to antigenic differences and may be a factor in outbreaks of contagious ecthyma in vaccinated goats.

Orf virus DNA vaccines expressing ORFV 011 and ORFV 059 chimeric protein enhances immunogenicity

BackgroundORFV attenuated live vaccines have been the main prophylactic measure against contagious ecthyma in sheep and goats in the last decades, which play an important role in preventing the outbreak of the disease. However, the available vaccines do not induce lasting immunity in sheep and goats. On the other hand, variation in the terminal genome of Orf virus vaccine strains during cell culture adaptation may affect the efficacy of a vaccine. Currently, there are no more effective antiviral treatments available for contagious ecthyma.ResultsWe constructed three eukaryotic expression vectors pcDNA3.1-ORFV011, pcDNA3.1-ORFV059 and pcDNA3.1-ORFV011/ORFV059 and tested their immunogenicity in mouse model. High level expression of the recombinant proteins ORFV011, ORFV059 and ORFV011/ORFV059 was confirmed by western blotting analysis and indirect fluorescence antibody (IFA) tests. The ORFV-specific antibody titers and serum IgG1/IgG2a titers, the proliferation of lymphocytes and ORFV-specific cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α) were examined to evaluate the immune responses of the vaccinated mice. We found that mice inoculated with pcDNA3.1-ORFV 011/ORFV059 had significantly stronger immunological responses than those inoculated with pcDNA3.1-ORFV011, pcDNA3.1-ORFV059, or pcDNA3.1-ORFV011 plus pcDNA3.1-ORFV059. Compared to other vaccine plasmids immunized groups, pcDNA3.1-ORFV011/ORFV059 immunized group enhances immunogenicity.ConclusionsWe concluded that DNA vaccine pcDNA3.1-ORFV011/ORFV059 expressing ORFV011 and ORFV059 chemeric-proteins can significantly improve the potency of DNA vaccination and could be served as more effective and safe approach for new vaccines against ORFV.

Rift Valley Fever in Ruminants, Republic of Comoros, 2009

Rift Valley Fever in Ruminants, Republic of Comoros, 2009

Severe Persistent Contagious Ecthyma Cases in Twin Goats

Severe Persistent Contagious Ecthyma Cases in Twin Goats

Phylogenetic analysis of Croatian orf viruses isolated from sheep and goats

BackgroundThe Orf virus (ORFV) is the prototype of the parapoxvirus genus and it primarily causes contagious ecthyma in goats, sheep, and other ruminants worldwide. In this paper, we described the sequence and phylogenetic analysis of the B2L gene of ORFV from two natural outbreaks: i) in autochthonous Croatian Cres-breed sheep and ii) on small family goat farm.ResultsSequence and phylogenetic analyses of the ORFV B2L gene showed that the Cro-Cres-12446/09 and Cro-Goat-11727/10 were not clustered together. Cro-Cres-12446/09 shared the highest similarity with ORFV NZ2 from New Zealand, and Ena from Japan; Cro-Goat-11727/10 was closest to the HuB from China and Taiping and Hoping from Taiwan.ConclusionDistinct ORFV strains are circulating in Croatia. Although ORFV infections are found ubiquitously wherever sheep and goats are farmed in Croatia, this is the first information on genetic relatedness of any Croatian ORFV with other isolates around the world.

Contagious ecthyma in Cuba: Evidence for a unique causative viral strain

Contagious ecthyma in Cuba: Evidence for a unique causative viral strain

Virological and clinico-pathological features of orf virus infection in experimentally infected rabbits and mice

Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts to establish animal models have yielded conflicting and non-reproducible results. We herein describe the characterization of ORFV infection and disease in rabbits and mice. A protocol of intradermal inoculation was employed to inoculate 10 8.5TCID 50/mL of ORFV strain IA-82 in the skin of ears, of the back and labial commissures. All inoculated rabbits presented a clinical course characterized by erythema, macules, papules/vesicles or pustules that eventually dried originating scabs. Local signs started around days 3 and 4 post-inoculation (pi) and lasted 3–10 days. Virus was recovered from lesions between days 2 and 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit milder clinical course occurred in 5/10 inoculated mice; virus was recovered from lesions from three animals. Inoculated lambs – used as controls – developed severe lesions of contagious ecthyma. VN tests performed at day 28pi failed to detect neutralizing antibodies in all inoculated animals. In contrast, convalescent rabbit sera were positive by ELISA at dilutions from 100 to 400. These results show that rabbits are susceptible to ORFV infection and thus may be used to study selected aspects of ORFV biology.

Histopatología de las infecciones víricas cutáneas más frecuentes

We describe the histopathological characteristics of viral skin infections. Herpes simplex virus and varicella-zoster virus produce an intraepidermal vesicle with variable degrees of epithelial necrosis. Typical findings include keratinocytes with ballooned nuclei with a ground-glass appearance and giant multinucleated keratinocytes. In the endothelial cells of the dermal blood vessels, cytomegalovirus produces large eosinophilic nuclear inclusions surrounded by a clear halo. Human herpes virus 8 is etiologically associated with Kaposi sarcoma. In its early stages, this tumor contains blood vessels with a fine endothelium passing through the dermal collagen bundles. In the plaque and nodular stages, the vessel lumens are more clearly visible and there is a progressive increase in the number of neoplastic spindle cells with a low degree of pleomorphism and atypia, and occasional mitoses. The infiltrate is made up of lymphocytes and plasma cells. Contagious ecthyma and milker's nodule give rise to an acanthotic epidermis with ballooned keratinocytes containing eosinophilic cytoplasmic viral inclusions. Molluscum contagiosum shows lobules of epithelium that open onto the epidermal surface and characteristic inclusion bodies. Acanthosis, papillomatosis, and hyperkeratosis are observed in common warts, with confluence of the epidermal ridges in the centre of the lesion and koilocytes.

A review of chemical, biological and fertility control options for the camel in Australia

Since their introduction to Australia in 1840 the one-humped camel, Camelus dromedarius, has gone from the colonist’s companion to a conservationist’s conundrum in the fragile arid ecosystems of Australia. Current management techniques are failing to curb present population growth and alternatives must be sought. This review assess the applicability of currently registered and developmental vertebrate pesticides and fertility control agents for camel control, as well as examining the potential usefulness of known C. dromedarius diseases for biological control. Not surprisingly, little is known about the lethality of most vertebrate pesticides used in Australia to camels. More has been published on adverse reactions to pharmaceuticals used in agriculture and the racing industry. An examination of the literature on C. dromedarius diseases, such as camel pox virus, contagious ecthyma and papillomatosis, indicates that the infections generally result in high morbidity but not necessarily mortality and this alone may not justify their consideration for use in Australia. The possibility exists that other undiscovered or unstudied biological control agents from other camilid species may offer greater potential for population control. As a long-lived species the camel is also not ideally suited to fertility control. Notwithstanding, anti-fertility agents may have their place in preventing the re-establishment of camel populations once they have been reduced through mechanical, biological or chemical means. Delivery of any generic chemical or fertility control agent will, however, require a species-tailored pathway and an appropriate large-scale deployment method. Accordingly, we put forward avenues of investigation to yield improved tools for camel control.

Health management practices and disease prevalence in dairy sheep systems in Argentina

A cross-sectional survey of 19 dairy sheep farms in Argentina was carried out with the purpose to know farm management, health practices, and occurrence and mortality of diseases. The survey comprised 40% of all sheep milking farms in Argentina. A questionnaire was conducted by way of personal interviews with sheep owners during farm visits. The proportions of farms reporting routine vaccination for clostridial diseases, contagious ecthyma, pneumonia and mineral and vitamin parenteral administration were 63%, 47.3%, 16.6% and 42.1% respectively. Regular treatment against lice was used in 37.5% of the farms, and 89.5% o the farmers treated against gastrointestinal nematodes (GIN). The mean number of GIN drenches per farm was 2.26±1.78 annually. In 68.4% of the milking flocks the California Mastitis Test was regularly done and 55.6% of the farmer managers had sampled their flocks once a year for Brucella ovis antibodies. During the pre-mating period respectively 68.4% and 50% of farmers clinically examined their rams and ewe for general health and teeth condition. The udders of ewes were frequently inspected at the start of each milking period. The most important parasite problems noticed were GIN (reported by 57.9% of farmers), lice (57.9%) and scabies (10.5%) and the most frequent infectious diseases were ecthyma (73.7%), pneumonia and other respiratory problems (57.9%), clinical mastitis (55.6%), clostridial diseases (36.9%) and foot lameness (35.2%). Photosensitivity (47.4%) and ruminal acidosis (42.1%) were reported as other frequent toxic or metabolic disorders. Owners mentioned that the mean lifespan or milk productive time per ewe was 4.5±1.4 years. Perinatal lamb mortality was 8.5% and the total flock mortality rates, above the first 24 h of life was 6.9%. The high rates of lamb mortality during the pre-weaning (10.3%) and post-weaning (5.9%) periods indicate that this problem, as well as the most prevalent diseases, should be the subject of further studies.

Identification and phylogenetic analysis of an Orf virus isolated from an outbreak in sheep in the Jilin province of China

This study investigated an acute outbreak of contagious ecthyma dermatitis that occurred in November 2008 in a herd of 180 small-tailed Han sheep in the Jilin province of China. The pathological findings of this case revealed severe vascular proliferation, viral cytopathic changes in keratinocytes by vacuolar degeneration, ballooning degeneration, and eosinophilic cytoplasmic inclusions, suggesting that the disease could have been caused by an Orf virus (ORFV) infection. Immunohistochemistry, indirect immunofluorescence (IFA), and transmission electron microscopy supported the diagnosis of ORFV infection. Finally, the pathogen of the disease was identified using polymerase chain reaction (PCR) and sequences of major envelope protein genes (ORFV 011(B2L) and ORFV 059(F1L)). The full-length ORFV 011(B2L) and ORFV 059(F1L) genes were cloned and sequenced. The nucleotide and amino acid sequences of ORFV 011(B2L) and ORFV 059(F1L) genes in these outbreaks were analyzed, and their phylogenetic trees were constructed. The phylogenetic studies of ORFV 011(B2L) genes and ORFV 059(F1L) genes showed that the ORFV-Jilin province isolate clustered in different branches and was closer to the ORFV-Mukteswar 67/04 isolate and the ORFV-OV/C2 isolate, respectively. To date, there is no report on the molecular characterization of any Orf virus with other isolates around the world in mainland China. Further, the above results may provide some insight into the genotype of the etiological agent responsible for the contagious ecthyma dermatitis outbreak in the Jilin province, and could also provide a comparative view of the central coding region genomics of parapoxvirus (PPV).

Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples

BackgroundOrthopoxvirus (OPV) and Parapoxvirus (PPV) have been associated with worldwide exanthematic outbreaks. Some species of these genera are able to infect humans and domestic animals, causing serious economic losses and public health impact. Rapid, useful and highly specific methods are required to detect and epidemiologically monitor such poxviruses. In the present paper, we describe the development of a nested-multiplex PCR method for the simultaneous detection of OPV and PPV species directly from exanthematic lesions, with no previous viral isolation or DNA extraction.Methods and ResultsThe OPV/PPV nested-multiplex PCR was developed based on the evaluation and combination of published primer sets, and was applied to the detection of the target pathogens. The method showed high sensitivity, and the specificity was confirmed by amplicon sequencing. Exanthematic lesion samples collected during bovine vaccinia or contagious ecthyma outbreaks were submitted to OPV/PPV nested-multiplex PCR and confirmed its applicability.ConclusionThese results suggest that the presented multiplex PCR provides a highly robust and sensitive method to detect OPV and PPV directly from clinical samples. The method can be used for viral identification and monitoring, especially in areas where OPV and PPV co-circulate.

Investigations into the Cause of Foot-and-Mouth Disease Virus Seropositive Small Ruminants in Cyprus During 2007

In 2007, serological evidence for foot-and-mouth disease (FMD) infection was found as a result of differential diagnostic testing of Cypriot sheep suspected to be infected with bluetongue or contagious ecthyma. Seropositive sheep and goats were subsequently uncovered on ten geographically clustered flocks, while cattle and pigs in neighbouring herds were all seronegative. These antibodies were specific for serotype-O FMD virus, reacting with both structural and non-structural (NS) FMD viral proteins. However, no FMD virus could be recovered from the seropositive flocks. FMD had not been recorded in Cyprus since 1964 and there has been no vaccination programme since 1984. Since all the seropositive animals were at least 3 years old and home-bred, it was concluded that infection had occurred approximately 3 years previously had passed un-noticed and died out spontaneously. It therefore appears that antibodies to FMD virus NS proteins can still be detected around 3 years after infection of small ruminants, but that virus carriers cannot be detected at this time. This unusual situation of finding evidence of historical infection in a FMD-free country caused considerable disruption and alarm and posed questions about the definition of what constitutes a FMD outbreak.

Detection and phylogenetic analysis of Orf virus from sheep in Brazil: a case report

BackgroundOrf virus (ORFV), the prototype of the genus Parapoxvirus (PPV), is the etiological agent of contagious ecthyma, a severe exanthematic dermatitis that afflicts domestic and wild small ruminants. Although South American ORFV outbreaks have occurred and diagnosed there are no South American PPV major membrane glycoprotein B2L gene nucleotide sequences available.Case presentationan outbreak of ovine contagious ecthyma in Midwest Brazil was investigated. The diagnosis was based on clinical examinations and molecular biology techniques. The molecular characterization of the virus was done using PCR amplification, cloning and DNA sequencing of the B2L gene. The phylogenetic analysis demonstrated a high degree of identity with ORFV strains, and the isolate was closest to the ORFV-India 82/04 isolate. Another Brazilian ORFV isolate, NE1, was sequenced for comparative analysis and also showed a high degree of identity with an Asian ORFV strain.ConclusionDistinct ORFV strains are circulating in Brazil. This is the first report on the phylogenetic analysis of an ORFV in South America.

Capripoxvirus identification by PCR based on P32 gene

Sheeppox virus (SPV) and goatpox virus (GPV) belong to the capripoxvirus genus of Poxviridae family. Sheeppox and goatpox along with contagious ecthyma (CE) are endemic diseases in Iran. Capripox laboratory conformation based on virological and serological techniques are time consuming, laborious and most of them of low specificity, because of close antigenic relationship between capripoxvirus and parapoxvirus. The aim of this study was to develop a capripoxvirus specific PCR assay for SPV & GPV identification on the basis of 390 bp fragment of P32 gene encoding capripoxvirus immunodominant antigen. PCR reaction was optimized using two reference strains of SPV & GPV and four field isolates in tissue culture supernatants. The identity of PCR product was confirmed by sequence analysis and the sensitivity of PCR was performed with 10 fold serial dilutions genomic LK cell DNA infected with capripoxvirus. This PCR was carried out on Twenty-nine biopsy samples from different organs of sheep and goats suspected to SPV & GPV against six biopsy samples infected with CE. Twenty-five SPV & GPV samples were positive and all of CE samples were negative. This PCR assay showed high sensitivity in detection of capripoxvirus DNA and good specificity in differentiation of capripoxvirus from parapoxvirus.

Diagnosis | Contagious ecthyma

Diagnosis | Contagious ecthyma

Development of a contagious ecthyma vaccine for goats

To identify a strain of contagious ecthyma virus from goats that possesses the appropriate characteristics for an effective vaccine for goats. 25 goat kids used for vaccine development and 100 goat kids used for evaluation of vaccine efficacy. 5 strains of contagious ecthyma virus were tested in a vaccination-challenge study to identify the best strain to be the seed strain for a contagious ecthyma vaccine. The vaccine derived from the chosen viral stain was tested at 2 concentrations for efficacy in a vaccination-challenge study. 2 of 5 viral strains induced moderate to severe scabs following infection, and 3 viral strains protected the goats from wild-type virus challenge following vaccination. Viral strain 47CE was selected as the seed source for the production of a contagious ecthyma vaccine because of the larger vaccine-to-challenge scab formation ratio. Vaccine 47CE protected all goat kids (48/48) following challenge with the wild-type contagious ecthyma virus; all goat kids (32/32) in the control group had scab formation following challenge with the wild-type contagious ecthyma virus, which indicated no protection following administration of vaccine diluent. A vaccine containing a caprine strain of contagious ecthyma virus used in goats appeared to provide the characteristics needed for an effective vaccine, including good scab production and protection from wild-type infection. This vaccine may potentially provide better protection for goats from contagious ecthyma than currently available vaccines labeled for sheep.

Recurrent localised cutaneous parapoxvirus infection in three cats

CASE HISTORY: Three cats were presented with single proliferative lesions affecting one foot, which failed to heal after medical treatment, and recurred despite surgical resection. PATHOLOGICAL FINDINGS: Histologically, the lesions were proliferative and papillary. There was marked acanthosis, rete peg formation, and compact orthokeratosis, with large numbers of bacteria in the orthokeratotic scale. Some biopsies had multifocal keratinocyte swelling of the stratum granulosum, and amphophilic intracytoplasmic inclusions were present in some of the swollen cells. The dermis consisted of a light fibrous stroma with marked capillary proliferation. Parapoxviruses were detected in the lesions of all cats by electron microscopic examination. PCR analysis detected orf virus (contagious ecthyma virus) in two cats, and orf virus was cultured from one cat. DIAGNOSIS: Parapoxvirus infection in cats CLINICAL RELEVANCE: Parapoxvirus infection should be considered as a differential diagnosis when dealing with proliferative, non-healing lesions on the feet of cats, especially cats in rural areas. The recovery of orf virus from a cat with typical poxvirus lesions extends the range of species affected by this virus.

Ectima contagioso em ovinos e caprinos no semi-árido da Paraíba

Descrevem-se dez surtos de ectima contagioso em caprinos e dois em ovinos no semi-árido da Paraíba. Os animais jovens foram os mais afetados, mas em oito surtos animais adultos também foram afetados, principalmente cabras lactantes que apresentavam lesões no úbere. Em nenhum dos rebanhos afetados havia antecedentes de vacinação contra ectima contagioso. Histologicamente a epiderme apresentava acantose com hiperplasia pseudoepiteliomatosa e com múltiplos focos de degeneração hidrópica formando vesículas ou pústulas. Inclusões intracitoplasmáticas eosinofílicas foram observadas em queratinócitos. Havia restos celulares necróticos formando crostas, hiperqueratose e microabscessos intracorneais ou extensas áreas ulceradas. Na derme observou-se infiltrado inflamatório mononuclear difuso. Também foi registrado um caso humano em um pesquisador que tinha administrado planta por via oral a um caprino que, posteriormente, apresentou lesões de ectima contagioso. Neste caso, também foram observadas lesões características em uma biopsia e o vírus foi identificado por microscopia eletrônica. Conclui-se que a doença é endêmica no semi-árido e que é necessária a vacinação sistemática dos rebanhos para diminuir os prejuízos econômicos causados pela mesma. A utilização de luvas para o manuseio de animais afetados é necessária para evitar casos humanos da enfermidade.