Main Subjects : Veterinary Virology

Isolation and identification of Circovirus in pigeon

Safwan Yousif Al-Baroodi; Mozahim yasen Al-Attar

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2020.126706.1364

The purpose of this study is first trial to detect of pigeon circovirus, so 1sr group include 100 cloacal swabs were collected 55 healthy and 45 ill pigeons, 36 yearlings and 64 adults, the 2nd group included organs was liver, spleen, bursa of Fabricius from 41 young pigeons 10-30 days old and bursa of Fabricius, liver, spleen from 28 dead in shell pigeon embryo in the 3rd group. DNA extracted from this samples and detection of virus DNA was attempt using polymerase chain reaction, after DNA amplification, the final products of the amplicon with 331 bp was cleared by using electrophoresis using agarose gel at concentration 2%. Results of viral DNA amplification were positive, which revealed as band in 331 bp the results showed that ill yearling pigeons recording high infectivity rate (66.7%)compare with healthy yearling pigeons and adult once, the bursa of Fabricius samples of dead yearling pigeons recorded high prevalence (36.58%) when compare with liver and spleen samples, DNA of pigeon circovirus high detected (60.71%) in bursa of Fabricius of dead in shell pigeon embryo. inconclusion pigeon circovirus affected the racing pigeon in Mosul, Iraq.

Development of in-house Taqman qPCR assay to detect equine herpesvirus-2 in Al-Qadisiyah city

Mohammed H. Al-Saadi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 365-371
DOI: 10.33899/ijvs.2019.126076.1229

EHV-2 is distributed in horses globally. It is clustered within gamma-herpesvirus subfamily and percavirus genus. EHV-2 infection has two phases: latent and lytic. In the later, EHV-2 mainly associated with respiratory and genital symptoms. However, in the quiescent phase of infection, EHV-2 stay dormant in the host till viral reactivation. Our previous study has showed that EHV-2 can be harboured by equine tendons, suggesting that leukocytes possibly carrying EHV-2 for the systemic dissemination. So far, numerous PCR protocols have been performed targeting the gB gene. However, this gene is heterogenic. Therefore, there is a need to develop a quantitative diagnostic approach to detect the quiescent EHV-2 strains. To do this, Taqman qPCR assay was developed to quantify the virus. This was performed by targeting a highly conserved gene known as DNA polymerase (DPOL) gene using constructed plasmid as a standard curve calibrator. The obtained results showed an infection frequency of 33% in which the EHV-2 load reached 6647 copies/100 ng DNA whereas the minimum load revealed as 2 copies/100 ng DNA. The median quantification was found as 141 copies/ 100 ng DNA. Establishment of a credited qPCR assay to quantify EHV-2 could be helpful in the control of the disease.

Prevalence of the bovine adenovirus type 3 by using direct fluorescent antibody technique in calves in Nineveh province

Abdulhakeem A. Sheet; Safwan Y. Al-Baroodi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 53-57
DOI: 10.33899/ijvs.2019.125476.1009

A total of 200 samples were collected from the calves for different ages from local and exotic breed by using nasal swabs, to investigate the prevalence of the bovine adenoviruses. The results showed that about 44% of the examined calves gave positive to immunofluorescence test, and the calves aged 6-9 months showed highly significant prevalence compare with other ages. The prevalence of virus infection in the exotic breed was 50.3% compared with local breed 22.2%. The calves with respiratory affections showed a higher prevalence from those which seem to be healthy. The current study concluded that the bovine adenovirus-type 3-virus has an effect on the breeding of calves in Nineveh Governorate.  

Diagnosis of reovirus infection in broiler breeders flocks by using PCR technique in Erbil province

Fanar Isihak

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 77-81
DOI: 10.33899/ijvs.2019.125469.1007

Avian reoviruses can infect birds without any clinical signs of infection, the infection may associate with different manifestations including viral arthritis/tenosynovitis and malabsorption syndrome. The objective of this study was to use advance methods representing by molecular methods (RT-PCR, RT-qPCR) in the diagnosis of ARV infection in broiler breeders' flocks. A 4 flocks of broiler breeders (ROSS breed) 39 weeks age with approximately10% morbidity rate due to Avian Reovirus (ARV). The clinical examination of 16 infected birds revealed unilateral lameness and swelling of hock joint. Blood samples were collected from wing vein of infected birds. Sera were tested for antibodies titer against ARV and Mycoplasma synoviae (MS). 5 of 16 positive samples were selected randomly for amplification by RT-PCR and RT-qPCR. The results showed in postmortem examination of infected birds, unilateral arthritis with visible joint lesions. Antibodies titer measured by ELISA in the sera of birds after 4 and 20 weeks of infection with ARV was positive and high. In RT- PCR1 of 5 samples gave positive reaction for amplification while in RT-qPCR all five samples gave positive results for amplification in comparison with +ve and -ve control.

Isolation and detection of reovirus from arthritis in chickens

Safwan Yousif Al-Baroodi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 59-63
DOI: 10.33899/ijvs.2019.125580.1093

In this study 70 samples were collected from 14-26 weeks old egg laying hens. Clinical signs of infected chickens characterized by lameness, swelling in hock joint. Samples include blood for preparation of serum also hock joints and tendon for virus isolation. Hyperimmune sera was prepared by injection of broiler chickens four times with vaccine strain Reo 1133R 0.2 ml subcutaneously in the neck. Samples were processed and prepared for virus isolation by using 6 days old embryonated chicken egg which was inoculated in yolk sac four passages. Lesions in egg embryo was recorded for each passage then the isolates were diagnosed by using neutralization test using convalescent and hyperimmune sera. Clinical signs of infected birds characterized by swelling and enlargement and edema of hock joint, postmortem lesions revealed swelling and injury in tendon, ulceration and erosions in cartilage and discoloration in synovial fluid, hemorrhage in the leg and yellow necrotic foci in the liver, the result of virus cultivation in embryonated chicken egg show dwarfism in growth, death of embryo with subcutaneous hemorrhage, initiated in 2nd passage and subsequent passages, this lesion increase in severity with progress of passages and with decrease in death time in hours and increase in titer of virus particles. The virus titer was decreased when neutralized by using neutralization test it gives 22 isolates were positive from 34 isolates.

Diagnosis and histopathological study of avian influenza virus-H5 (AIV-H5) in broiler farms

Fanar A. Isihak; Hana Kh. Ismail; Abed Alwaheed A. Wahid

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 101-107
DOI: 10.33899/ijvs.2019.125646.1120

This study was conducted for diagnosis and description of the pathological changes of AIV-H5 as the causative pathogen in Iraqi broiler farms. The current study was carried out on 84 broiler farms. Infected birds were tested for detection of the AIV infection from the tracheal swabs by rapid chromatographic AIV type A and H5 test kits. In RRT-PCR 8 samples (8 farms) of Trachea were selected to be tested by this assay. Samples of trachea, lung, and spleen from the dead birds with natural AIV-H5 infection were submitted for histopathological examination. seventy-two out of 84 farms tested for AIV-Type A gave positive results, and 58 out of 72 positives for type A-AIV gave a positive result for H5 antigen in a rapid chromatographic strip. The main gross lesions in the trachea of infected birds were severe congestion and hemorrhage. In the RRT-PCR assay, 8 out of 8 samples gave a distinct positive result for this test. The microscopic histopathological examination of infected tracheas showed obvious desquamation of lining epithelium with complete loss of cilia associated with congestion of blood vessels in lamina properia. Infected lungs revealed diffuse alveolar damage and severe multifocal vascular congestion. There was deposition of fibrinous material in the splenic tissue associated with the disappearance of the germinal centers. Thus, we concluded that AIV-H5 infection causes severe pathological and histopathological changes as a result of systemic infection. The RRT-PCR assay was highly sensitive and specific for the detection of highly pathogenic avian influenza virus subtypes.

Phylogenetic tree analysis study of bovine papillomaviruses type 1 based on L1 gene in Al-Qadisiyah governorate, Iraq

Khalefa Ali Mansour; Hassan Hachim Naser; Muthanna Hadi Hussain

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 151-155
DOI: 10.33899/ijvs.2019.125535.1057

Bovine fibropapilloma and papilloma occur in different parts of the skin of animals. Bovine Papillomavirus (BPV) is an oncogenic virus making benign tumor lesion of together mucosal and cutaneous tissue in cattle. In order to confirm the clinical diagnosis; the study planned to make the molecular detection of BPV (DNA) using Polymerase Chain Reaction (PCR) from skin lesions and the phylogenetic analysis. Thirty-eight samples of skin lesions were collected from cattle clinically suspected to be infected with bovine papilloma virus from herds in Al-Qadisiyah Governorate in 2016, the primary clinical diagnosis depended on the morphological appearance and features of the lesion. Deoxyribonucleic Acid (DNA) was extracted from skin lesions; the DNA was examined by PCR technique using specific primer to BPV-1 /L gene-1. Twenty-two samples out of 38 (57,9%), which were collected from different regions in Al-Qadisiyah Governorate, were positive. The sequences of four positive samples of DNA product amplification of (BPV) type-1, L1 gene confirmed the PCR results. These samples had the DNA presented in four accession numbers KY662042-1, KY662043-1, KY662040-1 and KY662041-1. This study proofed that cutaneous bovine papillomatosis related with BPV-1 infection in the cattle herds has affinity to solid skin rather than other epithelial and mucosal tissue.

First phylogenetic characterization of Pseudocowpox virus from cattle in Al-Qadisiyah province/ Iraq

Salah Mahdi Karim; Khalefa Ali Mansour; Ali Hassan Janabi; Nawras K. M. Al-Nakeeb

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 123-126
DOI: 10.33899/ijvs.2019.125525.1047

This study was initiated for the first time for identification, using sequencing and phylogenetic analyses, of pseudocowpox PCPV that inhabit dairy cows in Al-Qadisiyah province, Iraq. Scab sampling was performed to obtain specimens from udder and teats of 18 affected cows. Initially, a polymerase chain reaction (PCR) method was followed to target a 408-bp piece of the GM_CSF/IL-2 inhibition factor gene (GIF) that belongs to PCPV. Then, the PCR products were sent out to partial sequencing of the GIF gene. The results of the PCR have indicated the presence of the virus in only 3 out of 18 samples. When the sequences were studied using phylogeny, the results have revealed that one of our PCPV strains has a close matching with some of the world strains such as from New Zealand. While two of the current study strains have clustered together with a strain from Finland. The results of our study confirm the presence of the PCPV in dairy cows that induces milker’s nodules.

Molecular diagnosis and genetic relationship of foot and mouth disease virus serotype Asia1/Basne/Sul/2015

Jeza Muhamad Abdul aziz; Salih Ahmed Hama; Hawre Kamel Faraj

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 67-73
DOI: 10.33899/ijvs.2019.125519.1041

Foot and Mouth Disease (FMD) is the most economically important viral-induced livestock disease worldwide. From April to May of 2015, tongue epithelial tissue samples were collected from 36 cattle in six villages, which share the border with Iran. Samples were screened using RT-PCR to amplify a conserved region in the VP1 gene, and phylogenetic tree analysis was performed based on the VP1 nucleotide sequence results. Furthermore, the nucleotide sequence was converted to an amino acid sequence in order to detect similarities between the studied samples and those previously published in GenBank (NCBI). Epidemically, based on the amino acid residues, genetic similarity, and amino acid substitutions, the VP1 nucleotide sequences were determined to be close to a novel group, group VII, with 94% identity. The VP1 amino acid sequence analysis revealed a close relationship to the Asia/BAL/PAK/iso-2/2011 isolate (Accession no. JX435109), with 95.7% identity, which is not significantly different. Analysis of the studied samples revealed that the FMDV serotype Asia1 causing the outbreak in the Basne district belonged to group VII, which was introduced from the Balochistan province of Pakistan through illegal movement of animals from this region.

Molecular identification of peste des petits ruminants virus in wild goat and domestic small ruminants by real-time -PCR technique in Erbil-Iraq

E.P. Candlan; F.P. Khoran; L. Hana

Iraqi Journal of Veterinary Sciences, 2017, Volume 31, Issue 1, Pages 51-54
DOI: 10.33899/ijvs.2017.126710

In July 2010 outbreak was occurred in wild goat in Barzan, Sherwin mizzen and Mergasur in Kurdistan Region- Iraq. There were over 2700 deaths (both young and adult) during the period of July 2010 to October 2011. Based on the clinical signs and post-mortem findings, the involvement of peste des petits ruminants virus (PPRV) was suspected. This was confirmed by Real Time PCR technique using TaqMan®probes for the detection of Peste des petits ruminants. The results of Real-Time PCR for the 9 sample taken from 9 Wild goat there are 6 sample positive and 3 sample negative and 76 sample from domestic ruminants (sheep and goat) 63 samples was negative for PPR. This result confirms the diagnosis domestic ruminants in the region are routinely vaccinated with an attenuated vaccine based on the ‘Nigeria/75/1’ strain of PPRV.

Immunosuppressive effect of Marek's disease virus in broiler

Mozahim Yasen Al-Attar

Iraqi Journal of Veterinary Sciences, 2005, Volume 19, Issue 1, Pages 1-6
DOI: 10.33899/ijvs.2005.37262

تمت دراسة التأثیر المناعی المثبط للاصابة التجریبیة بفایروس مرض میرک فی فروج اللحم على مستوى المناعة المتکونة عند تلقیح حیوانات التجربة بلقاح مرض نیوکاسل وقد تم الکشف عن مستوى الأضداد المناعیة باستخدام اختبار تثبیط التلازن الدموی، حیث أظهرت نتائج هذا الاختبار انخفاض معیار الأضداد المتکونة ضد مرض نیوکاسل فی المجموعة الأولى المصابة تجریبیاً بفیروس مرض میرک والتی جرى تلقیحها بلقاح نیوکاسل بالمقارنة مع المجموعة الثانیة والتی اعطیت لقاح نیوکاسل فقط دون احداث الاصابة التجریبیة بفیروس مرض میرک، کما استخدم اختبار التحدی للتأکد من مستوى الحمایة التی أحدثها لقاح نیوکاسل حیث تبین أن هذا اللقاح أعطى حمایة بنسبة 50% فی المجموعة الأولى بینما کانت تلک النسبة 90% فی المجموعة الثانیة عند استخدام فیروس مرض نیوکاسل الضاری کجرعة التحدی لکلتا المجموعتین. وهذا یدل على أن الإصابة بفیروس مرض میرک فی الدواجن له تأثیر سلبی على مستوى المناعة التی یحدثها لقاح مرض نیوکاسل فی الدواجن.