Effect of a Topical Formulation on Infective Viral Load in Lambs Naturally Infected with Orf Virus

Delia Lacasta,1 Ramses Reina,2 Marta Ruiz de Arcaute,1 Luis Miguel Ferrer,1 Alfredo Angel Benito,3 Maria Teresa Tejedor,4 Irache Echeverria,2 Hector Ruiz,1 Silvia Martinez Cardenas,1 Peter Andrew Windsor5 1Animal Pathology Department, Instituto Agroalimentario de Aragón-IA2 (Universidad de Zaragoza-CITA), Veterinary Faculty of Zaragoza, Zaragoza, 50013, Spain; 2Instituto de Agrobiotecnología (CSIC-Gobierno de Navarra), Mutilva, 31192, Navarra, Spain; 3EXOPOL S.L, San Mateo de Gállego, Zaragoza, Spain; 4Anatomy, Embryology and Animal Genetics Department, CIBER CV (Universidad de Zaragoza-IIS), Veterinary Faculty of Zaragoza, Zaragoza, 50013, Spain; 5Sydney School of Veterinary Science, The University of Sydney, Camden, NSW, 2570, AustraliaCorrespondence: Delia LacastaFacultad de Veterinaria de Zaragoza, C/Miguel Servet 177, Zaragoza, 50013, SpainTel +34 609676727Email dlacasta@gmail.comIntroduction: Orf is a highly contagious eruptive viral disease of the skin and mucosa of sheep and goats. Although vaccination with live or attenuated orf virus is the preferred option for disease control, the vaccine is unavailable in many countries. Treatment of orf lesions involves standard hygiene and in numerous cases, management of presumptive secondary infections with antibiotics, increasing risks of antimicrobial resistance (AMR). The wound dressing formulation Tri-Solfen® containing two local anaesthetics (lignocaine and bupivacaine), adrenaline and an antiseptic (cetrimide) in a gel formulation was developed for pain relief in sheep undergoing surgical husbandry procedures in Australia. Recently, TS therapy was found to reduce suffering and enhance recovery in cattle and buffalo with oral and skin lesions due to foot-and-mouth disease (FMD) virus infection. It was noted that TS has a low pH and is potentially viricidal, potentially aiding disease control.Methods: One-month-old lambs (n=14), naturally infected with orf, were recruited from a farm during a natural outbreak of the disease. The animals were selected at the early stages of the infection and randomly divided into two cohorts: Group A (n=11) treated with the topical wound gel formulation (TS); and Group B (n=3) an untreated control group. Swabs were obtained before treatment (T0) and on days one (T1), 3 (T2) and 5 (T3) post-treatment, then submitted to direct DNA extraction with real-time PCR quantification, plus incubation with primary tissue cultures from ovine skin fibroblasts (OSF) and T-immortalized goat embryonic fibroblasts (TIGEF).Results: Although no significant differences were found in the clinical progression of the lesions and PCR quantification (p=0.722) between these small cohorts, there was a significant difference (p< 0.05) in reduction in infective viral load between the groups when assessed in OSF cell cultures between T0 and T3.Conclusion: These preliminary findings suggest that treatment of early stage lesions with this TS may reduce the infective viral load present in orf lesions.Keywords: sheep, contagious ecthyma, wound formulation, local therapy