Investigation of Prenatal Infection in a Calf of a Rabid with Natural Delivery

Kuduz virusu ensefalomiyelitise neden olan son derece nöyrotropik bir virusdur. Gebe on yaşındakibir hoştayn inek durgunluk, böğürme, inkoordinasyon, dilin dışarı sarkması, kuyruk hareketlerindeazalma, salivasyon, kamburlaşma, aşırı duyarlılık, saldırganlık ve takiben felç gibi klinik belirtilergösterdi. İnek bir ay öncesinden bir köpek tarafından ısırılmıştı. O ölü bir erkek dana doğdu. İnekdoğumdan bir gün sonra öldü. İnek ve buzağının kafatası kemikleri açılarak beyinleri çıkartıldı.Klinik ve otopsi gözlemleri sadece kuduzdan şüphelendirdi. Beyinler kuduz yönünden teşhislaboratuvarında test edildi. İnekte kuduz hastalığı teşhisi yapıldı. Kuduz virusu, fare inokulasyonuile izole edildi. Rabies virus antijenleri ve spesifik virus genleri flöresan antikor test (FAT) vereverze transkription polimeraz zincir reaksiyonu (RT-PCR) ile belirlendi. Buzağının cerebellum vecornu ammonislerinden yapılan FAT testinde negri cisimciklerine rastlanmadı. Buzağı tükrük bezi,lenf düğümleri, böbrek, kalp kası, akciğer, karaciğer, barsak, dalak, spinal cord ve beynin cornuammonis ve cerebellum dokularından FAT, RT-PCR ve fare inokulasyonu yapıldı. Dokulardankuduz virusu izole edilemedi. Kuduz virusu antijenleri ve spesifik virus genlerine rastlanmadı.İnekten elde edilen pozitif RT-PCR amplifikasyon ürününden kısmi dizin analizi yapıldı veGenBank'da TR/EL/753/2009/cattle KM058759 kodu ile yayınlandı. RT- PCR amplifikasyonürününün virus nükleoproteini N geni kısmi 0-510 bp dizin analizinin NCBI programındadeğerlendirilmesinde izolatın Rabies Virus (RABV) olduğu doğrulandı. RABV' nin filogenetik analizibelirlendi. Dizinin filogenetik analizi sonucuna göre, Türkiye ve Türkiye'nin Ortadoğuda bulunankomşu ülke izolat dizinleri ile identik olduğu belirlendi. Bu araştırma, kuduz inekten doğan birbuzağıda rabies virusunun, bulunmaması spesifik bir vakada, transplesantal bulaşmanıngözlenmediğini göstermektedir.

Doğal Şartlarda Kuduz Bir İnekten Doğan Buzağıda Prenatal Enfeksiyonun Araştırılması

Rabies virus is a highly neuronotropic virus that causes encephalomyelitis. A pregnant 10-years-oldHolstein cow was presented with yawning, bellowing, incoordination, decreased feeling in hindlimbs, loss of tail movement, drooling saliva, "cud-dropping", inability to stand, hypersensitivity, andaggression followed by paralysis. The cow had been bitten by a dog from a month ago. It calved adead male calf. Than the cow died a day later after parturition. The skull of the cow and calf wereopened and brains were removed. Clinical and post mortem observation may only lead to asuspicion of rabies. Brains were examined for rabies in the diagnostic laboratory. Rabies wasdiagnosed in the cow. Rabies virus isolated in mouse inoculation. Rabies virus antigens andspecific virus genes were detected by fluorescent-antibody test (FAT) and reverse transcriptionpolymerase chain reaction (RT-PCR). But rabies was not diagnosed in the calf. Post mortemexamination of the dead male calf was made. There was congestion in the lungs and petechialhaemorrhages in the endocardium. Ammon's corn, cerebellum, spinal cord, umbilical cord, salivarygland, lymph nodes, kidney, myocardia, lung, liver, intestine and spleen of dead calf were collectedin a necropsy room. Fluorescent antibody test, hemi nested RT- PCR and the mouse inoculationtest were made for this tissues. Rabies virus not isolated in the mouse inoculation test. Rabiesvirus antigens and specific virus genes were not found by FAT and RT-PCR. Partial nucleotidsequence was made from the cow brain sample and published in GenBank withTR/EL/753/2009/cattle KM058759 code. According to the partial RT-PCR amplicon resultconfirmed that the isolate is Rabies Virus (RABV) by NCBI Programme. Phylogenetic analysis ofRABV identified 0-510 sequence of rabies virus nucleoprotein N gene. As a result, nucleotidsequence of the isolate were found as identical with the isolates obtained from province isolates, aswell as related with isolates sequences from neighboring countries of Turkey in the Middle East.This research demonstrated the absence of rabies virus in the fetus of a rabit cow thus, in thisspecific case, the transplacental transmission did not ocur.

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1. Fauquet EM, Mayo MA, Maniloff J, Desselgerger U, Ball LA. Virus taxonomy: Classification and nomenclature of viruses. Eighth report of the International Committee on the Taxonomy of Viruses. Virus Taxonomy. 8th Edition, San Diego: Academic Press, 2005.
2. Heaton PR, Johnstone P, Mc Elhinney LM, et al. Heminested PCR assay for detection of six genotypes of rabies and rabies related viruses. J Clin Microbiol 1997; 35: 2762-2766.
3. Meslin FX, Kaplan MM, Koprowski H. An Overview of Laboratory techniques in diagnosis and prevention of rabies and in rabies resarch. In: Meslin FX, Kaplan MM, Koprowski H. (Editor). Laboratory Techniques in Rabies, 4th Edition, Geneva: WHO, 1996: 9-27.
4. Office International des Epizooties (OIE). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Paris: OIE, 2008.
5. Woldehiwet Z. Rabies: Recent developments. Res Vet Sci 2002; 73: 17-25.
6. Iehl'e C, Dacheux L, Ralandison S, et al. Delivery and follow-up of a healthy newborn from a mother with clinical rabies. J Clin Virol 2008; 42: 82-85.
7. Müller-Holve W. Leitritz H, Knorr E. Early development of a child following rabies of the mother during pregnancy. Infection 1977; 5: 49-50.
8. Spence MR, Davidson DE, Dill Jr GS, Boonthai P, Sagartz JW. Rabies exposure during pregnancy. Am J Obstet Gynecol 1975; 23; 655-656.
9. Swende TZ, Achinge GI. Clinical rabies in pregnancy with delivery of a live fetus: A case report. Niger J Med 2009; 18; 1: 114-115.
10. Howard DR. Transplacental transmission of rabies virus from a naturally infected skunk. Am J Vet Res 1981; 42: 691-692.
11. Martell MA, Montes FC, Alcocer R. Transplacental transmission of bovine rabies after natural infection. J Infect Dis 1973; 127: 291-293.
12. Hemachudha T, Laothamatas J, Rupprecht CE. Human rabies: A disease of complex neuropathogenetic mechanisms and diagnostic challenges. Lancet Neurol 2002; 1: 101-109.
13. Hemachudha T, Kamolvarin N, Tirawatnpong T, et al. Anteand postmortem diagnosis of rabies in man and animal by PCR using nested primers. Neurol 1992; 42: 370.
14. World Health Organization Expert Committee on Rabies Eight Report, WHO Technical Report Series 1992.
15. Un H, Johnson N, Vos A, et al. Genetic analysis of four human rabies cases reported in Turkey between 2002 and 2006. Clin Microbiol Infect 2009; 15: 1185-1189.
16. Tamura K, Dudley J, Nei M, Kumar S: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007; 24: 1596-1599.
17. Mims CA. Vertical Transmission of Viruses. Microbiol Rev 1981; 45: 2267-286.
18. Viazhevich VK. Delivery of a normal infant from a mother in incubation period of rabies. Zh Microbiol Epidemiol Immunobiol 1957; 28: 105-106.