Isolation and identification of bovine herpes virus -1 (BHV-1) from semen of foreign breeds bulls

Madbouly, H. M.; Tamam, S. M.; Abd-El-Gaid, A. M.

doi:10.21608/jvmr.2008.77831

Cited by 5 publications

(6 citation statements)

References 4 publications

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“…Typical pocks were observed onto CAM on subsequent passages. The characteristics pock lesions were also observed by Madbouly et al , Madbouly and Hussein [ 14 , 19 ] when they collected semen sample from dairy herds having genital form of infection. The result of present study showed large and small pock lesions were on CAM.…”

Section: Discussionsupporting

confidence: 58%

“…All samples were collected in the capped sterile plastic vials and transported on ice to the laboratory and processed as the method by Madbouly et al . [ 14 ] as quickly as possible, after freezing and thawing several times (3-5 cycles) semen was diluted in ratio of 1:10 with fetal bovine serum (OIE) then centrifuged at 3000 rpm for 10 min. The suspension of these specimens was filtered through 0.45 μm size filters and used for virus isolation.…”

Section: Methodsmentioning

confidence: 99%

“…For virus isolation, 11-day-old embryonated chickens eggs were inoculated through CAM route [ 14 ]. Five serial passages were performed for obtaining the good titer of virus and the presence of BHV-1 was confirmed by AGID test [ 15 ].…”

Section: Methodsmentioning

confidence: 99%

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Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Samrath¹,

Shakya²,

Rawat³

et al. 2016

Vet World

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Aim:Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin-Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test. Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test. Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as "bunch of grapes" at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test. Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test.

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Section: Discussionsupporting

confidence: 58%

Section: Methodsmentioning

confidence: 99%

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Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Samrath¹,

Shakya²,

Rawat³

et al. 2016

Vet World

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“…It was used for the detection of antibody against BHV1 in cattle and buffaloes serum and was used according to [8,17]. Diluted to1/100 then each sample was added to the micro-plate wells.…”

Section: Indirect Elisamentioning

confidence: 99%

Serological and Molecular Identification of Infectious Bovine Rhinotrachetitis Virus Isolation and Adaptation in Embryonated Chicken Eggs

Zeedan¹,

Abdalhamed²,

Ghazy³

et al. 2018

J Antivir Antiretrovir

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Infectious bovine rhinotracheitis virus (IBRV) is a major pathogen in livestock animals and had led to significant economic losses to the industrial production worldwide. However, IBR symptoms are not life-threatening. The present study was achieved to make seroprevalence and isolation of bovine herpes virus type (BHV-1) from nasal and ocular swabs samples of suspected cows and buffaloes, identification by serological and polymerase chain reaction (PCR). Total of 380 blood samples were collected from suspected 287 cattle and 93 buffaloes in different districts at Beni-suef and El-Fayoum governorates in winter 2017. Nasal and ocular discharge swabs were collected from cattle and buffaloes with clinical respiratory signs (Nasal discharge, cough, lachrymal discharge with or without mild diarrhea and higher body temperature). A total of 106 (27.89%) samples were positive by indirect ELISA and the positivity of 80 (27.87%) samples from cattle and 26 (27.96%) samples from buffaloes located at different centers at Beni-Suif and El-Fayoum governorates. Virus was isolated from nasal and ocular discharges swabs samples and it was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cells. Virus-infected CAM showed congestion, edematous vacuole, thickening small foci ranged from 2 to 3 mm in diameter, scattered on CAM membrane and MDBK cell line inoculated blind serial passages at 3rd passage showed characteristic of cytopathic effect (CPE). Identification of virus isolated on CAM and infected cell culture fluid gave precipitation against positive specific anti-BHV-1 immune serum by AGPT and clear blue zone by Dot ELISA, absent of pock lesion in pock reduction test (PRT), and confirmed by PCR with product size of 175 bp. Finally BHV-1 virus was isolated from nasal and ocular discharges of cattle and buffaloes but the further extensive study still need for clear final classification by phylogenic analysis.

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“…Namun demikian, pertumbuhan virus tersebut pada embrio ayam tidak menimbulkan kematian dan perubahan patologi pada embrio ayam. Menurut Madbouly et al (2008), bahwa isolasi BHV-1 yang berasal dari swab preputium menghasilkan bentukan lesi pock yang spesifik pada membrana korioallantois. Karakteristik dari pock tersebut adalah berukuran besar dengan tepi opak rata dan adanya area sentral nekrosis yang berwarna keabuan.…”

Section: Metode Penelitianunclassified

"Deteksi Bovine Herpesvirus-1 Secara Immunohistokimia pada Membran Korioallantois Telur Ayam Berembrio (IMMUNOHISTOCHEMISTRY DETECTION OF BOVINE HERPESVIRUS-1 IN CORIOALLANTOIC MEMBRANE OF CHICKEN EMBRYONATED EGG)"

Kristianingrum

Tabbu²,

Sutrisno³

et al. 2015

jveteriner

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Infectious Bovine Rhinotracheitis (IBR) adalah penyakit yang disebabkan oleh Bovine Herpes virus-1 (BHV-1) pada sapi dengan gejala klinis depresi, anoreksia, pembengkakan vulva, kemerahan pada vestibula, pustula pada mukosa yang bisa melanjut menjadi ulserasi. Berdasarkan penelitian sebelumnya virus IBR yang berasal dari swab nasal ini dapat ditumbuhkan dalam membrana korioallantois telur ayam berembrio. Penelitian ini bertujuan untuk meneguhkan diagnosis infeksi virus IBR pada sapi yang ditumbuhkan dalam telur ayam berembrio sebagai pengganti kultur sel. Lima sampel swab nasal dari sapi yang didiagnosis positif menderita IBR dengan peneguhan secara Polymerase Chain Reaction dan kultur sel, diinjeksikan pada membran korioallantois telur ayam berembrio umur 9-10 hari. Pengamatan lesi dilakukan 3-5 hari setelah inokulasi. Penanaman kembali (pasase) dilakukan sebanyak tiga kali. Lesi karakteristik berupa pock dapat diamati pada membran korioallantois dengan ukuran 5-7 mm, berbentuk bulat, tepi opak dengan area sentral nekrosis. Selanjutnya lesi pock diproses jaringan untuk pewarnaan Hematoksilin dan Eosin (HE) dan immunohistokimia. Berdasarkan hasil pewarnaan dengan menggunakan HE ditemukan bentukan badan inklusi intranuklear, dan vakuolisasi epitel sel membran. Hasil pewarnaan immunohistokimia menunjukkan reaksi positif terhadap antibodi BHV-1 pada sel epitel membran korioallantois. Berdasarkan hasil penelitian ini dapat disimpulkan bahwa telur ayam berembrio dapat digunakan sebagai media untuk deteksi virus penyebab IBR. Kata-kata kunci : bovine herpes virus-1, lesi pock, membran korioallantois ABTRACT Infectious Bovine Rhinotracheitis (IBR) is caused by Bovine Herpes virus-1 in the cattle. The clinical signs demonstrate depression, anorexia, swelling of the vulva, redness of the vestibule, pustule and ulcer on the vaginal mucosal. Based on previous research, IBR virus from the nasal swab could be grown in chorio-allantoic membrane of embryonated chicken eggs. This study aim was to confirm whether IBR virus in cattle could be grown in embryonated chicken eggs as a substitute for cell culture. A total of five nasal swab samples from the cows that were positive for IBR infection (diagnosed by Polymerase Chain Reaction and cell culture) were inoculated on the chorio-allantois membrane of embryonated chicken eggs. Observation of lesions performed at 3-5 days after inoculation. Re-inoculation (passage) was done three times. Pock characteristic lesions were observed on the corioallantoic membrane with the size of 5-7 mm, rounded shape, opaque edge, with necrosis in the central area. Furthermore, pock lesions were processed for hematoxylin and eosin staining and immuno-histochemistry. The result of hematoxylin and eosin staining showed that the formation of intranuclear inclusion bodies and vacuolization of the epithelial cell of membrane was observed. Immuno-histochemistry staining showed positive reaction for antibodies against BHV-1 in the epithelial cells membrane. In conclusion, embryonated chicken eggs could be use...

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Isolation and identification of bovine herpes virus -1 (BHV-1) from semen of foreign breeds bulls

Cited by 5 publications

References 4 publications

Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Serological and Molecular Identification of Infectious Bovine Rhinotrachetitis Virus Isolation and Adaptation in Embryonated Chicken Eggs

"Deteksi Bovine Herpesvirus-1 Secara Immunohistokimia pada Membran Korioallantois Telur Ayam Berembrio (IMMUNOHISTOCHEMISTRY DETECTION OF BOVINE HERPESVIRUS-1 IN CORIOALLANTOIC MEMBRANE OF CHICKEN EMBRYONATED EGG)"

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