Determination of the Infectivity of Cryopreserved Theileria annulata Sporozoites in Tick Derived Stabilates Iran Ak-93 Strain, by In Vivo and In Vitro Methods.

BACKGROUND: The protozoan parasite Theileria annulata is the causative agent of tropical theileriosis in cattle. Vaccination is recommended by administration of attenuated schizont-infected cell lines. The expected protective immunity post-vaccination can be demonstrated by challenge test through inoculation of highly virulent infective sporozoites. The aim of this study was to produce Hyalomma anatolicum anatolicum tick infected with T. annulata (local strain) for preparation of tick-derived sporozoite stabilates for molecular characterization and infectivity test assay. METHODS: A local T. annulata strain was used for experimental infection of calves. A field isolate of H. a. anatolicum was isolated, laboratory-reared and infected by blood-feeding on Theileria infected above-mentioned calves. The infectivity of calf, tick and prepared stabilate were confirmed by clinical signs of theileriosis, microscopic inspection, RT-PCR and in vitro cell culture. RESULTS: The tick stabilate was prepared and cryopreserved in liquid nitrogen. The infectivity of the tick stabilate was verified by in vivo bioassay, in vitro cell culture infection, microscopic inspection in salivary glands and RT-PCR assay. The in vitro produced cell line in this study was characterized by T. annulata Cytochrome b gene analyzing. CONCLUSION: The infectivity of a new prepared tick-derived sporozoite stabilate was confirmed in susceptible calves; by microscopically, post mortem, tick microscopic and molecular assays. Moreover, naïve PBMCs were transformed and proliferated by T. annulata infected tick stabilate to immortal T. annulata schizont infected cell line. The potent infective sporozoite tick derived stabilate could be used for vaccine efficacy and challenge test as well as in vaccine development.

Peste des Petits Ruminants Virus in Vulnerable Wild Small Ruminants, Iran, 2014-2016.

In 2014-2016, >1,000 wild goats and sheep in 4 northern and central provinces of Iran died from peste des petits ruminants virus (PPRV) infection. Partial nucleoprotein sequencing of PPRV from 3 animals showed a close relationship to lineage 4 strains from China. Control measures are needed to preserve vulnerable ruminant populations.

West Nile virus monitoring in migrating and resident water birds in Iran: are common coots the main reservoirs of the virus in wetlands?

A molecular and serological study was carried out to determine the West Nile virus (WNV) status in different species of wild water birds. From 2003 to 2007, samples were collected from 519 birds representing 26 different species in Iran. Out of 519 serum samples tested for WNV antibodies, 78 (15%) were positive when tested using virus neutralization and immunofluorescence. Antibodies of WNV were detected in 71 out of 131 common coot (Fulica atra) samples. In comparison, only 7 out of 388 birds that were belonged to 25 other species of water birds revealed positive results. For most Anatidae species, no positive duck in serological tests was found. Further, no WNV viral RNA-positive samples were found in this study. Results of this investigation provide important information about the prevalence of WNV in wild water birds in Iran and indicate the potential role and importance of common coots in ecology of WNVs.

Avian influenza virus monitoring in wintering waterbirds in Iran, 2003-2007.

BACKGROUND: Virological, molecular and serological studies were carried out to determine the status of infections with avian influenza viruses (AIV) in different species of wild waterbirds in Iran during 2003-2007. Samples were collected from 1146 birds representing 45 different species with the majority of samples originating from ducks, coots and shorebirds. Samples originated from 6 different provinces representative for the 15 most important wintering sites of migratory waterbirds in Iran. RESULTS: Overall, AIV were detected in approximately 3.4% of the samples. However, prevalence was higher (up to 8.3%) at selected locations and for certain species. No highly pathogenic avian influenza, including H5N1 was detected. A total of 35 AIVs were detected from cloacal or oropharyngeal swab samples. These positive samples originated mainly from Mallards and Common Teals.Of 711 serum samples tested for AIV antibodies, 345 (48.5%) were positive by using a nucleoprotein-specific competitive ELISA (NP-C-ELISA). Ducks including Mallard, Common Teal, Common Pochard, Northern Shoveler and Eurasian Wigeon revealed the highest antibody prevalence ranging from 44 to 75%. CONCLUSION: Results of these investigations provide important information about the prevalence of LPAIV in wild birds in Iran, especially wetlands around the Caspian Sea which represent an important wintering site for migratory water birds. Mallard and Common Teal exhibited the highest number of positives in virological and serological investigations: 43% and 26% virological positive cases and 24% and 46% serological positive reactions, respectively. These two species may play an important role in the ecology and perpetuation of influenza viruses in this region. In addition, it could be shown that both oropharyngeal and cloacal swab samples contribute to the detection of positive birds, and neither should be neglected.