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Original Article the Role of the Gray Wolf in Rabies Transmission in Iran and Preliminary Assessment of an Oral Rabies Vaccine in This Animal

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Original Article the Role of the Gray Wolf in Rabies Transmission in Iran and Preliminary Assessment of an Oral Rabies Vaccine in This Animal J Med Microbiol Infect Dis, 2017, 5 (3-4): 56-61 Original Article The Role of the Gray Wolf in Rabies Transmission in Iran and Preliminary Assessment of an Oral Rabies Vaccine in this Animal Alireza Gholami1*, Siamak Massoudi2, Majid Kharazian Moghaddam2, Mahmoud Ghazi Marashi2, Mahyar Marashi2, Rouzbeh Bahsar1, Ahmad Fayaz1, Maryam Fazeli1, Firouzeh Farahtaj1, Nader Howaizi1, Mohammad-Reza Shirzadi3 1National Reference Center for Rabies, Pasteur Institute of Iran, Tehran, Iran; 2Department of Environment, Wildlife Diseases Group, Wildlife Bureau, Tehran, Iran; 3Department of Zoonoses, Center for Communicable Disease Control, Ministry of Health and Medical Education, Tehran, Iran Received Feb 20, 2018; Accepted Feb 24, 2018 Introduction: In Iran, 95% of animal bites are attributed to domestic dogs and cats, while one-third of rabies victims are from wildlife. The wolf is one of the primary vectors of wildlife in the country. This study was aimed to assess the impact of wildlife on rabies transmission and to evaluate the efficacy of an oral rabies vaccine (ORV) in gray wolf (Canis lupus pallipes). Methods: We studied the incidence of wolf attacks by analyzing a 10-year period of data (2000 to 2009) available in the Ministry of Health and Medical Education, the Veterinary Organization and Pasteur Institute of Iran. In the next step, five captive gray wolves were fed by Raboral V-RG® vaccine baits (Merial®, part of Boehringer Ingelheim, Germany) and monitored for rabies neutralizing antibodies over 20 months by rapid fluorescent focus inhibition test (RFFIT). Results: Our analysis showed that 55% of animal bites occurred in rural areas and the remaining in urban areas. The most rabid wolves were from Fars province in southern Iran. The vaccine baits were palatable for the hungry wolves. Seroconversion with antibodies titers ≥0.5 IU/ml occurred after 12 weeks and maintained for 78 weeks indicating potential protection against rabies after receiving a single ORV bait. Conclusion: Our results suggested that a single dose of vaccination with V-RG®, though initially designed for foxes and raccoons, can protect the gray wolves against rabies as well. J Med Microbiol Infect Dis, 2017, 5 (3-4): 56-61. DOI: 10.29252/JoMMID.5.3.4.56 Keywords: Wolf, Rabies, Iran, Oral rabies vaccination, Anti-rabies antibodies. INTRODUCTION Rabies is an acute progressive viral encephalitis and one been identified in Iran’s wildlife with the disease spillovers of the most critical zoonotic diseases worldwide with the to domestic animals in these areas [9, 10]. Previous studies fatality rate of almost 100% [1]. The disease is commonly showed that dogs and foxes in the north and wolves in the transmitted through the bite of rabid animals. The west and northwest of Iran were the most important vectors etiological agent of the disease is an enveloped single- of the disease. Dogs were the most frequent potential stranded negative-sense RNA virus belonging to the source of exposure to rabies (62%) during 1993-1994 and Rhabdoviridae family and the genus Lyssavirus 2008-2009. In Khorasan and Kerman provinces in the east (Order Mononegavirales). Members of this family occur on and south of Iran, wild rabies was more critical in all continents except Antarctica [2]. Carnivores are the comparison to other regions [11, 12]. In Ilam province, main reservoirs of the virus in nature and dogs play a western Iran, dogs were responsible for the majority of significant role in the transmission of rabies to humans in animals bites (89.2%) during 1999-2009, followed by cats most developing countries. Whereas, wild animals are (5%), wolves (0.7%), jackals (0.6%), foxes (0.3%), and mainly responsible for the dissemination of rabies in other animals (4.2%) [13]. In Tehran, most of the animal developed countries [3]. Some of these countries have bites were inflicted by dogs (65.9%), followed by cats controlled sylvatic cycle of rabies by oral vaccination. (25.44%), ferrets (3.98%), monkies (1.52%), hamsters Transmission of rabies from wildlife to domestic animals (1.41%), and other animals (1.84%) [14]. and humans remains a severe public health concern in many Vaccination of wild animals and dogs can prevent the countries [4]. In addition to human deaths, rabies is circulation of the rabies virus in the wildlife and its responsible for substantial cattle losses and pose a threat to rare carnivore species [5, 6]. In northern provinces of Iran, the majority of rabies occurs in livestock kept in barns [7, *Correspondence: Alireza Gholami 8]. Rabies, as an endemic disease, circulates among both National Reference Center for Rabies, Pasteur Institute of wild and domestic animals of Iran. Annually, the national Iran, No. 69, Pasteur Ave, Tehran, Iran, 1316943551 reference center for rabies, Pasteur Institute of Iran, Email: [email protected] confirms several hundred rabid animals. In addition to the dogs, wild carnivores such as foxes, jackals, and wolves are Tel: +98 (21) 64112227 Fax: +98 (21) 64112813 vectors of rabies in wildlife. Until now, 62 rabies foci have http://jommid.pasteur.ac.ir Gholami et al. transmission to domestic animals and humans as well. some oral vaccine baits for rabies control in wildlife [20]. Annually, about 300-400 thousands owned dogs receive a For example, according to the EU requirements, the SAG2 vaccination by veterinary clinics in Iran. However, this baits were efficient for vaccination of the red fox and approach does not cover stray dogs. The leftovers and raccoon dog, and Raboral V-RG® has been widely used to garbage attract stray dogs and wild animals resulting in an control rabies in foxes and raccoons [21, 22]. This increased risk of human exposure to animal bites. This recombinant anti-rabies vaccine, due to using a live virus, is condition also increases the possibility of crossbreeding of only recommended for wild animals. The recombinant dogs and wolves. More than 160,000 animal bites by vaccinia virus is stable at elevated temperatures and can be various animals are reported annually, of which 10.8% are delivered orally. Different reports showed the elimination attributed to wolves (CDC, Ministry of Health and Medical of sylvatic rabies by using Raboral V-RG® in United States, Education (MOHME) of Iran). In some developing Canada, and several other countries [23]. countries, preventive vaccination against rabies is History of the wolf associated rabies in Iran recommended for people and especially for children who demonstrates the aggressive behavior and excessive live in endemic areas. In most endemic regions of the world, physical power of this animal in inflicting deep and 99% of rabies transmissions to humans are due to the dog numerous injuries to the victim's body. Hence, the rabid bites. Hence, mass vaccination of dogs is considered as the wolf has long been considered as one of the most dangerous most efficient way to control human rabies [15]. In Iran, carriers of the disease in wildlife of Iran [24]. The current dogs and cats are responsible for almost 95% of the bites, study has analyzed the role of wild animals in the whereas wild animals are responsible for more than one- transmission of rabies in Iran and the ability of Raboral V- third of human rabies deaths. RG® as an ORV in the production of neutralizing antibodies The sylvatic cycle of rabies preserves the virus among against rabies virus in the captive gray wolves (Canis lupus wildlife and maintains the likelihood of rabies reemergence pallipes). [16]. Elimination of rabies in wildlife and prevention of the disease transmission from wildlife to domestic and MATERIAL AND METHODS companion animals in endemic areas can be achieved by ® mass vaccination campaigns [17]. In developed countries, Vaccines. We used Raboral V-RG vaccine baits for attempts are directed to control rabies in wildlife, and oral oral vaccination. Each bait block (size: ~1 ¼” x 1 ¼”) 7.7 rabies vaccination (ORV) has received considerable contained 2 ml of recombinant vaccinia virus (10 plaque attention as a promising and socially acceptable approach. forming units per ml) encoding the rabies virus surface The oral bait vaccines can be distributed over extensive glycoprotein, coated by fishmeal. For comparison, Rabisin areas in a short period. Until now, several strains of rabies vaccine was used for vaccination through parenteral route. ® vaccine baits have been developed and applied to the The commercial company of Merial manufactures both wildlife worldwide [18, 19], and the WHO has approved vaccines. Fig. 1. Location of Koohsar town (shown by the red square) in Alborz province of Iran, where the wolves were kept, and the vaccination study was performed J Med Microbiol Infect Dis 57 2017 Vol. 5 No. 3-4 Wolf rabies and oral vaccination in Iran Animals. Five adult healthy captive gray wolves (Canis reversed with 200 mcg/kg in 0.4 ml/10 kg of Atipamezole lupus pallipes) including two males and three females with hydrochloride. A microchip was implanted subcutaneously an average weight of 36 and 26 kg, respectively were used in each animal for further examinations. in this study. A group of four wolves was fed by vaccine Statistical analysis of data. Statistical analysis of the baits, and a single wolf received vaccination through data obtained from the Veterinary Organization and parenteral route. Production of neutralizing antibodies was Department of Environment of the Islamic Republic of Iran monitored for 16 months. This interventional study was were performed by the SPSS software version 9 (SPSS Inc., performed at wildlife rehabilitation center of Issar Chicago, USA). Statistics were given in the form of mean recreation and cultural complex, Koohsar county, Karaj, ±SD along with the median. Iran (Fig. 1). Vaccine Administration. Before feeding with baits, the animals were starved for 24 h, and then each animal in the RESULTS test group was allowed to feed on one bait.
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