Keywords : PCR

Molecular characterization of extended spectrum cephalosporin resistant Escherichia coli isolated from dogs

Ihsan M. Ahmed; Sumaya Y. Aldabbagh; Dhyaa M. Jwher

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2020.127032.1441

Shepherd dogs have been implemented in the transmission and distribution of many threatening pathogens. The presence of extended-spectrum-cephalosporin resistant Escherichia coli (ESCR E. coli) in dog feces can constitute a significance risk to human health due to transmission of antibiotics resistance from dogs to humans, other animals and the surrounding environment. Therefore, in this study, phenotypic and molecular characterization of fecal ESCR E. coli were investigated in shepherd dogs accompanied sheepherders in urban areas. Sixty-seven fresh fecal samples were collected from shepherd dogs from different regions of Mosul city. Bacteriological examination of ESCR E. coli was done using MacConkey agar with cefotaxime followed by subsequent PCR confirmation of the CTX-M gene using specific primers and molecular characterization using specific primers directed to CTX-M-1, 2 and 9 groups. The results of bacterial examination showed successful confirmation of ESCR E. coli which has been isolated from fecal samples of shepherd dogs 58.2% (39/67). In addition, detection of CTX-M gene was confirmed in 53.7% (36/67) of E. coli isolates. Furthermore, molecular characterization of CTX-M gene revealed the presence of only one genotype belongs to CTX-M-1. However, both of CTX-M-2 and CTX-M-9 genotypes were not detected in this study. This study concluded that shepherd dogs have an essential role in carrying and spreading of ESCR E. coli especially in urban regions.

Detection of CTX-M gene in extended spectrum β-lactamases producing Enterobacteriaceae isolated from bovine milk

Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 397-402
DOI: 10.33899/ijvs.2020.126909.1412

Extended spectrum β-lactamases producing Enterobacteriaceae (ESBL-E) have emerged recently as the main cause that facilitates the spreading of antibiotic resistance worldwide. Due to its composition and nutritive values, raw cow milk is vulnerable to bacterial contamination from different sources, especially ESBL-E. Accordingly, present study aimed to detect the ESBL-E in the raw milk of healthy cows. 80 raw cow milk samples were collected from unorganized farms and cows belong to individual owners and investigated for the presence of ESBL-E with the main focusing on CTX-M type. The bacterial isolation was performed using selective MacConkey agar plus cefotaxime (MC+), while PCR was used to confirm the species of the isolated bacteria and presence of CTX-M gene. The results showed that 28.75%(23/80) samples were ESBL-E positive and distributed as following, 82.61%(19/23) were pure E. coli isolates, 4.35%(1/23) was pure K. pneumoniae isolate and finally, 13.04%(3/23) were mixed of both E. coli and K. pneumoniae isolates. Moreover, the total number of positive ESBL-E was 26 isolates with the majority of them were belong to E. coli and recorded 84.61%(22/26), while K. pneumoniae was recorded less 15.39%(4/26). Additionally, the CTX-M gene was successfully identified in all ESBL-E positive isolates by using PCR, including E. coli and K. pneumoniae isolates. The results of this study assert the importance of raw cow milk as a potential source of ESBL-E that might be transmitted to humans.

Molecular differentiation of Thysaniezia (Helictometra) giardi and Moniezia species based on 18s rRNA gene in small ruminants

Monyer A. Alfatlawi; Yahia Kh. Ismail; Mansour J. Ali; Azhar C. Karawan; Israa N. Ibadi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 105-108
DOI: 10.33899/ijvs.2020.126407.1313

This study was conducted to investigate Anoplocephalidia Cestoda in sheep and goat and evaluate the 18s rRNA to genetically differentiate the genera of this family. Sixty sample tapeworms were collected from small intestines of 30 sheep and 30 goats from different slaughterhouses in Al-Najaf and Al-Qadisiyah provinces, during September, 2016 to February, 2017. Based on polymerase chain reaction (PCR) and 18s rRNA gene partial sequencing (18sGPS) methods used, tapeworm infection of sheep and goat’s intestines was 32.9% and 31.4%, respectively. The partial gene sequencing of the 18S rRNA gene showed two closely related isolates of M. benedeni which are aligned distinctly to an NCBI isolate of the same species from China. For T. giardia, the outcomes of the phylogenetic analysis unveiled three distinct local isolates which were similar to an NCBI database isolate from China. The current data ensure the importance of the molecular techniques in differentiating between Thysaniezia (Helictometra) giardi and Moniezia species that were identified for their presence in the small intestines of sheep and goats.

Molecular identification of new circulating Hyalomma asiaticum asiaticum from sheep and goats in Duhok governorate, Iraq

Shameeran Ismael; Lokman Tayib Omer

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 79-83
DOI: 10.33899/ijvs.2020.126330.1298

Because there was no such study done on identification of tick species by PCR technique in in Duhok Governorate, therefore present study was done to identify tick species by using molecular study by using of 16S rRNA and DNA sequencing. About 1000 ticks were collected from both sheep and goat, form Duhok Governorate including: Barwaria, Zakho, Sumeil, Mangeshik, Sersing, Shekhan and Akre, between May and June 2016, between April and June 2017. The result found during this study were six species under two genera of the hard ticks were identified by molecular study and sequencing including: three species were under the genus Hyalomma and three species were under the genus Rhipiciphalus that infect small ruminants in Duhok governorate from these species a new species under the Hylomma genra (Hyalomma asiatium asiaticum) with accession number (MN594484), was first time reported in Duhok governorate. Also phylogenetic tree was constructed depend on the 16S rRNA.

Study of Staphylococcus aureus isolated from the mouth of canary

Saba A. Hussein

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 301-304
DOI: 10.33899/ijvs.2019.125937.1192

The study was done for described genotypically characterize of Staph. aureus isolated from the oral cavity of canary birds in Mosul city using polymerase chain reaction technique which was achieved by amplifying of the thermonuclear nuc gene specialized with Staph. aureus. Sixty birds were examined from variable ages of both sexes from different regions of Mosul city for the period of 1/5/2018-1/6/2019 was carried out. The results indicate that 35 samples gave Staph. aureus with the percentage of 58.4%. These isolates are positive for pigmentation of mannitol salt agar, hemolysis on blood agar, catalase and coagulase-positive, gram staining and oxidase negative. PCR technique indicate that all 35 isolates were positive for the nuc gene and produce amplicon of 166 bp. These results considered positive and it is very specific for bacterial isolates of staph aureus as well as may be used for strain isolation, characterization, and differentiation from other types of bacteria.

Molecular detection and seroprevalence of Toxoplasmosis in free range local chickens (Gallus domesticus) in Duhok province, Iraq

Farhad B. Mikaeel; Adel T. Al-Saeed

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 247-252
DOI: 10.33899/ijvs.2019.125885.1173

Toxoplasmosis is a cosmopolitan zoonotic parasitic disease of mammals and birds; human infection occurs through consumption of raw or undercooked meat. Little was known about the infection rate of T. gondii among free range local chickens (Gallus domesticus) in Duhok province. Therefore, the present study was carried out to determine the infection rate in Duhok province by using ELISA (IgG) and conventional PCR. A total of 368 blood samples were collected from free range local chickens distributed in five different areas of Duhok province during the period from November 2016 to March 2017. The collected blood samples were from different sexes (hens and cocks) and from different age groups (less than 6 months and older than 6 months). The data found that the total infection rate was (84 / 368) 22.8% by ELSIA. The presence of the infection was confirmed by PCR and DNA sequencing. In this study, there were differences from area to area in the infection rates, the highest rate was reported in Semel district at 33.7% which was significantly (p

Molecular detection of Cryptosporidium parvum in chicken in Al-Diwaniya province

Noor I. Jarad

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 441-445
DOI: 10.33899/ijvs.2019.126159.1249

This study were conducted in Al-Diwaniya province, in south Iraq during the period from Februaryto July 2019 to determine the rate of infection of Cryptosporidium parvum in domestic chicken, study the effect of some epidemiological factors such as sex and months on the rate of infection, addition to the molecular identification of Cryptococcus parvum by amplification HSP70 gene by conventional PCR. Number of collected fecal sample was 210 from domestic chicken and stained by Ziehl-Neelsen stain. The results of the microscopic examination showed that 108(51.4%) out of 210 fecal samples were infected with Cryptosporidium spp. The statistical analysis founded no marked difference in prevalence of infection between sexes. Significant difference was recorded between infection rate during the months of the study and higher prevalence of infection rate was observed in March 11.9%, while lowest infection rate was observed in July 5.23%. and June 5.23%. Genomic DNA was extracted from 108 fecal samples and HSP70 gene for C. parvum was amplified by PCR. PCR technique is showed that out of 108 fecal samples 21.3% were positive for Cryptosporidium parvum.

Detection and pathogenicity of Listeria monocytogenes in common carp (Cyprinus carpio) fish in Baghdad, Iraq

Nagham M. Al-Gburi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 311-316
DOI: 10.33899/ijvs.2019.125980.1205

A few reports are available for detection of L. monocytogenes in fish in Iraq, however, the current study was undertaken to investigate the potential role of Listeria spp. in common carp fish in Baghdad province, Iraq. A total of fresh thirty raw common carp (Cyprinus carpio) were purchased from fish sellers of various local markets in Baghdad city from (December 2017 to March 2018) The viscera was removed aseptically, the bacterial isolation and identification was conducted by a conventional culture method using Listeria selective media, biochemical tests and Vitek 2 for gram-positive. Pathogenicity of isolates was studied in vivo by inoculating mice with bacterium. Targeting virulence associated genes was used to detect the virulence and to confirm the L. monocytogenes isolates. The isolates were tested for antimicrobial susceptibility by disk diffusion method for 12 antibiotics. The results revealed that 6.66% of L. monocytogenes were identified from common carp fish viscera and the isolates were pathogenic in mice. L. monocytogenes virulence associated genes were detected in both isolates, while L. innocua virulence associated gene (Lin0372) was detected in one of the two isolates. The isolates were resistant to 7 out of 12 antibacterial drugs including tetracycline, ampicillin, methicillin, cefixime, oxacillin, cefotaxime and penicillin G. The results suggest that presence of L. monocytogenes in fish may have a serious role in public health hygienic in humans.

Detection of Mycobacterium paratuberculosis in raw cow’s milk using polymerase chain reaction (PCR) technique

Ihsan M. Ahmed; Raad A. Al-Sanjary; Haiffa H. Alkazaly

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 83-86
DOI: 10.33899/ijvs.2019.125556.1075

Paratuberculosis or Johne’s disease is a chronic debilitating disease mainly infects ruminants and caused by Mycobacterium paratuberculosis. Previous serological studies in Mosul city confirm the presence of positive reactants for paratuberculosis in cattle. However, culture methods to confirm the disease need a long incubation period and also special media. Raw cow’s milk is considered as potential source for transmission of M. paratuberculosis in cows’ herds. Accordingly, this study aimed to detect the presence of M. paratuberculosis specifically in the raw cow’s milk using polymerase chain reaction (PCR) technique as a rapid, sensitive and reliable method. A total of 50 samples of raw cow’s milk were collected from cows suffering from emaciation and unresponsive to antibiotic treatment. All the samples were subjected to DNA extraction and direct amplification PCR. The results showed that 3 (6%) out of 50 milk samples were positive for M. paratuberculosis. This is the first study in Mosul city that confirms the presence of M. paratuberculosis in raw cow’s milk using PCR technique. In conclusion, raw cow’s milk could be an important source for M. paratuberculosis infection in dairy cows, and also PCR technique could be helpful in rapid diagnosis of paratuberculosis.

Detection of Shigella in raw bovine milk by polymerase chain reaction

Noor Soulieman; Aemaan Al-Mariri; Faizah Al-Atrash

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 9-16
DOI: 10.33899/ijvs.2019.125758.1146

Shigella is an intracellular bacterium can infect both human and animal. Its species especially Shigella dysenteriae cause shigellosis worldwide, with 165 million cases of severe bloody diarrhea and mucoid feces. The aim of this study was to find a rapid, sensitive and specific method for screening Shigella in raw bovine contaminated milk. For this goal, 70 samples of milk collected in sterile containers for isolating of Shigella and culturing it on selective media to identify and characterize its morphology, biochemical and molecular characteristics. This study was compared between three different DNA extraction techniques for polymerase chain reaction (direct DNA extraction using a kit, alkaline DNA extraction, and filtrated milk). Our results showed that PCR was able to detect Shigella in 15 out of 15 cases after the milk samples filtered. In other words, the filter technique can be used to detect Shigella in contaminated milk.

Investigate the Toxoplasma gondii infection in the consumed beef in Al-Diwaniyah province

Farah M. Sakban; Noman N. A’aiz

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 95-99
DOI: 10.33899/ijvs.2020.164336

Toxoplasmosis is very important zoonotic disease in the world cause by an obligate intracellular protozoan parasite called Toxoplasma gondii can infect human and all warm-blood animals, beef consider from most important source for infection with T. gondii and there is no really data and study about the rate of the infection in beef in Al-Diwaniyah province, so for this reason the aim of this work was designed. A total of 300 samples which collected from heart, tongue, muscles, of 100 slaughtered beef of local and imported cattle, throughout the period from September 2017 to May 2018, initially examined microscopically for searching on T. gondii bradyzoites then all suspected samples was subjected to conventional PCR technique through B1gene amplification to confirm the infection, in addition to analyzed the recorded data for each sample to determine the effect of some factors on prevalence of infection like organ, season and animal age. Out of 300 tested samples only 53 were confirm positive T. gondii DNA. The infection in local beef was higher (22%) than in imported (13.5%), while there is no difference in infection among different examined organs. Regarding to effect of some factors, the autumn season recorded highest rate of infection with significant differences rather than others seasons in both local and imported beef, whereas, age appears with no effect on infection. The local cattle meat is riskier than the imported due to the higher rate of infection with T. gondii, and the animal age cannot affect on the infection rate, in comparing with the season which play role in this rate.

Microscopic identification, molecular and phylogenetic analysis of Babesia species in buffalo from slaughter house in Al-Najaf city of Iraq

Rashaa Ateaa; Mansour Alkhaled

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 251-258
DOI: 10.33899/ijvs.2019.162882

Babesia is one of hemoprotozoan parasite transmitted by arthropod vectors which responsible for causing of Babesiosis disease in bovine worldwide. The present study was designed for microscopic identification, molecular, and phylogenetic analysis of Babesia species in buffalo from slaughter house in Al-Najaf city of Iraq. The study performed in three months of summer season (August into September 2017) and animals ages and sex were included in this study. The direct microscopic prevalence results were show highest prevalence of haemoprotozoa prevalence at Babesia sp. 45.74%. The prevalence of Babesia sp. related to animal sex, were show in male 43.48% and female was 52%, with non-significant differences. The Prevalence of Babesia sp. related to age were show 12.50%, 92.86% and 30% in young, adult and old age respectively with significant differences (P<0.05). The prevalence of Babesia sp. related to month of study were show. 28.57%, 62.50% and 42.86 in August, September and October respectively and with non-significant differences. Molecular study results were based on PCR and DNA sequencing method by phylogenetic tree analysis (MEGA 6.0) and NCBI-BLAST Homology Sequence Identity to differentiation Babesia species typing. The Babesia species prevalence results were show identified two Babesia species, high prevalence of Babesia bovis (38.30%) were closed related to NCBI-Blast Babesia bovis (HQ264126.1) with homology sequence identity 97-100% and Babesia bigemina 7.45% were closed related to NCBI-Blast Babesia bigemina (KU206291.1) with homology sequence identity 95-99%, then 43 Babesia species includes (B. bovis and B. bigemina) were submitted into NCBI-Genbank and provided accession numbers (MH503811-MH503853). In conclusion, this study concluded that Phylogenetic tree and homology sequences identity was show accurate in differentiation of Babesia species, and these species can be isolated at from local water buffalo from slaughter house in Al-Najaf city, of Iraq.

Detection of Mycoplasma gallisepticum and Mycoplasma synoviae by using of cultural and PCR technique

N.A. Jafar; Bashar Noomi

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 469-473
DOI: 10.33899/ijvs.2019.125484.1016

Laboratory methods are essential for the diagnosis of Mycoplasmal infection. There are three laboratory approaches are essential for the diagnosis of Mycoplasmal infection in chicken including direct methods by culture method and polymerase chain reaction, and indirect methods by detection of Mycoplasmal antibodies by serological tests. This study aimed to detection of Mycoplasma by culture and PCR technique. Two hundred seventy-six samples were collected from infected adult boiler chicken in Salah Al-din province which suffering from respiratory signs and /or joint infection, 202 respiratory and 74 articular samples. According to the results of culture, Mycoplasma isolated in rate of 35.1% (36.6% from respiratory samples and 31.1% from articular samples). The sensitivity of culture was 100%, while the specificity of culture was 97.9% when comparing with PCR results. The current study concluded that the respiratory infection was more than articular infections, and Mycoplasma gallisepticum more distributed than Mycoplasma synoviae among chickens.

Isolation and identification of Salmonella typhimurium bacteria with detection of type-1 fimbriae coding gene by polymerase chain reaction (PCR) technique

Zeena Saleh; B.M. Al-Muhana; Kh. Hamdan; M.S. Jawad; S.F. Klaif

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 195-199
DOI: 10.33899/ijvs.2019.162961

Two hundred faeces sample were collected from cattle with different age and sex in Al- Diwaniyah Province. The study was conducted in the period between November 2016 and November 2017. Salmonella typhimurium bacteria identified by routine methods such as culturing on selective media, biochemical test and agglutination test using monovalent and multivalent antisera. PCR was can detection type-1 fimbriae gene coding for fimC of Salmonella typhimurium. Results showed that Salmonella isolates were 14.5% in the bovine fecal samples. Also, the serotyping of isolates by using monovalent and polyvalent antisera revealed that all Salmonella isolates in cows were S. typhimurium. The PCR technique was used for detection of type-1 fimbriae coding gene by specific primer for fimC gene. All S. typhimurium isolates in cows appeared to be contained this gene show one distinct band MW.289 bp when electrophoresed on agarose gel. The results of this score indicated that the PCR technique potentate a loud specify in the disclosing of S. typhimurium especially the serotype that encoded to fimC gene type-1 fimbriae isolated from cows in comparison to other routine diagnostic tests.

Serodiagnosis of Toxocariasis by ELISA test using anti- T. canis IgG antibodies in stray dogs compared to PCR

Noor Jarad; A.K. Abbas; N.N. Aἀiz

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 367-370
DOI: 10.33899/ijvs.2019.163081

Toxocara (T.) canis is a nematode parasite of canines; belong to the Ascarididae family, which accidentally infected humans. Puppies expel the eggs with the feces from the fourth week of the life cycle. This study is the first study in Iraq for detection seroprevalence in stray dogs and extended from January to September 2017. Our study was aimed to investigate the seroprevalence of T. canis infection in stray dogs from different areas in the Al-Diwaniya province, Iraqto detection of specific IgG antibodies to T. canis compared to Conventional PCR technique with the effect of the risk factor. One hundred of the blood sample and one hundred of a faecal sample of same dogs after shooting were studied usingindirect ELISA test and PCR. The result revealed that 71% of the dogs had a seropositive result for this parasite by ELISA test. Dog age is an important factor and affects seroprevalence, were shown that positive rate in adult dogs was more 83.05% than the young dogs 53.65%, while no significant between dogs according to sex. PCR technique showed 58% of dogs were positive forinternal transcribed spacer 1 (ITS1) ribosomal RNA. The sensitivity and specificity of ELISA test was 79 and 40% respectively.

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.

Sequencing-based phylogenetic-study of Babesia spp detected in tick tissues in Al-Diwaniyah province, Iraq

Marwa Saleem Hajeel; Monyer Abdulameir Abd Alfatlawi

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 9-12
DOI: 10.33899/ijvs.2019.125512.1034

Our study purpose was to investigate the evolution of Babesia spp isolated from tissues of ticks that were found on 150 cows in Al-Diwaniyah province, Iraq. To fulfill the required purpose, sampling of 10 ticks was performed from each infested cow. These obtained ticks were morphologically recognized first, and then they were introduced to Lab investigation that was started with crushing the tick tissues to extract the genomic DNA of the Babesia spp. The DNA was then applied to polymerase chain reaction (PCR) method to recognize the amplification of the region that is related to the 18S rRNA gene. The resulted-amplified products were sequenced for the purpose of confirming and doing the phylogenetic analyses. Here, our study has demonstrated 2 different species according to the results of the sequencing and the phylogenetic analyses of the tested Babesisa species. These 2 species are SP1 and SP2. When the phylogenetic tree was built up, the results showed that SP1 and SP2 are closely related to Babesia bovis (HQ264126.1), an isolate from Texas, USA. Our study indicates interesting and valued data that could be used to study various aspects of the tick, Babesia species, and their control in Al-Diwaniyah City, Iraq.

Molecular study of Anaplasma marginale parasite in carrier cattle in Al-Nasiriyah city

N. R. Al-Kasar; M. M. Flayyih; A. D. Al-Jorany

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 2, Pages 299-301
DOI: 10.33899/ijvs.2019.153867

To detect Anaplasma marginale among carrier cattle by using polymerase chain reaction (PCR) technique, 64 blood samples, fromhealthy cows in abattoir of Al-Nasiriyah city were collected from June till August, 2017 in this study. By targeting MAR1bB2 gene with the molecular weight of approximately 265 bp, Anaplasma marginale were detected in18 samples (28.125%). One of these positive sample was recoded in National Center for Biotechnology Information, NCBI; Gene Bank.

Polymorphism of growth hormone gene in the artificial insemination result of Madura cattle with Limousin semen as a reference for genetic selection

B. Utomo; E. Safitri

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 1, Pages 113-118
DOI: 10.33899/ijvs.2018.153832

Research on genetic polymorphism of growth hormone (GH) and receptor growth hormone (rGH) has not been done in crossbred of Limousin cattle, so it is interesting to be examined. Blood samples were taken from 14 Madura calves were artificially inseminated with Limousin cement. DNA amplification is done by using Polymerase Chain Reaction (PCR) method, Restriction Fragment Length Polymorphism (RFLP) method to determine the genotype. DNA sequencing was done to determine nucleotide sequences of GH unit genes. The results showed that identification of GH and rGH gene polymorphisms was done by breaking DNA fragments from 432 and 298 bp in Madura and Limousin cattle (Madrasin) ie, L and V alleles have a frequency of 0.67 and 0.33 for the GH gene, respectively. This proves that the crossed-breeding of Madrasin have V allele that is not owned by the Madura cattle. While in the rGH gene, the A allele is 0.92 and the G allele is 0.08, with the frequency of the A allele larger than the G allele. This research concluded: that GH and rGH undergo changes on polymorphisms in Madrasin cattle can be used as a basis for selection.

Use molecular techniques as an alternative tool for diagnosis and characterization of Theileria equi

M.A. El-Seify; N.M. Helmy; N.M. Elhawary; Sh.S. Sorour; A.M. Soliman

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 1, Pages 5-11
DOI: 10.33899/ijvs.2018.153787

The purpose of this study was to determine the prevalence of clinical, subclinical and chronic infection with the equine parasite T. equi in some Egyptian localities (Cairo and Giza governorates). A panel of 396 equine blood samples representing 141 horses, 250 donkeys and 5 mules was collected from equines during the period from April 2015 to March 2016 using microscopic examination and conventional PCR. Microscopically a twenty two (5.56%) of 396 were positive for T. equi merozoites that appeared as small rounded, pyriform shaped and maltase cross shaped merozoites. Among 8/141(5.67%) horses and 14/250 (5.60%) donkeys were found to have positive for T.equi. A one hundred blood samples (45 horses, 50 donkeys and 5 mules) selected randomly were also examined by PCR. The results of PCR showed 30/100(11/45 (24.4%) horses, 18/50 (36%) donkeys and 1/5 (20%) mule) were positive for T.equi. When the sequenced PCR amplicons (n=3) were aligned to the reference nucleotide sequences of T. equi accessed in Genbank, the horse isolate showed insertion of Thymine (T) base at position 23 and substitution of Thymine (T) base with Cytosine (C) base at position 91, while the donkey and mule isolates have no alterations when compared to the reference sequences. The phylogenetic analysis showed that the sequenced PCR isolates belonged to T. equi. The obtained sequences were deposited in the GeneBank database under accession numbers MF192854, MF192855 and MF192856.

Prevalence and molecular studies on Echinococcus equinus isolated from necropsied donkeys

A.Y. Desouky; N.M. Helmy; Sh.S. Sorour; M.M. Amer

Iraqi Journal of Veterinary Sciences, 2017, Volume 31, Issue 2, Pages 101-106
DOI: 10.33899/ijvs.2017.145605

In the present study, forty donkeys of different ages and sexes at Giza Zoo, Egypt were investigated between October 2015 and September 2016 for the presence of hydatidosis disease. Hydatid cysts were detected in the livers of 10% of the examined donkeys and these cysts had a fertility rate 100%. Female donkeys were infected with cysts more than males and all infected donkeys were old aged with no cases of infection were detected in young or adult donkeys. Using molecular tools, the DNA extracted from cysts that had been isolated was subjected to PCR amplification, using synthesized oligonucleotide primers, and these were constructed to target the 299 bp within the (ND2) gene, which is considered to be specific for the Echinococcus equinus genotype. The sequenced PCR products showed homology to E.equinus (G4 or horse strain genotype). These results can be used in future to pursue the epidemiological status of the causative strain of hydatidosis in equines at the study area.

Phylogenetic study of Theileria lestoquardi based on 18SrRNA gene Isolated from sheep in the middle region of Iraq

M.J.A. Alkhaled; N.N. A'aiz; H.H. Naser

Iraqi Journal of Veterinary Sciences, 2016, Volume 30, Issue 2, Pages 27-32
DOI: 10.33899/ijvs.2016.121380

Theileriosis is parasitic infection causes by obligate intracellular protozoa of the genus Theileria. T. lestoquardi is the most virulent species in sheep and goats which causes a severe disease with a high morbidity and mortality rate. In this study the phylogenetic relationships between two local isolate of T. lestoquardi and nine T. lestoquardi global isolates as well as Babesia ovis out-group isolate were analyzed using the 18S rRNA gene sequence. The multiple sequence alignment analysis and neighbor joining phylogenetic tree analysis were performed by using ClustalW multiple sequence alignment online based analysis of 1098bp 18S rRNA gene was amplified by polymerase chain reaction. Phylogenetic analysis results of these gene sequences revealed that T. lestoquardi local isolates were closely related to T. lestoquardi Iran isolate (JQ917458.1) and two Iraq Kurdistan isolates (KC778786.1 and KC778785.1) more than other countries. This study represents the first report on the use of molecular phylogeny to classify T. lestoquardi obtained in Middle Region of Iraq.

Using species-specific PCR technique to detect Toxoplasma gondii in broiler chickens

R.A. Al-Sanjary; T.H. Hussein

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue 2, Pages 53-56
DOI: 10.33899/ijvs.2012.67452

Two groups of broiler chickens were used in this study. One was reared under typical conditions at the animal house of Veterinary Medicine College/Mosul University- Iraq, while the other group was reared under common commercial farm conditions. Fifty and 80 birds from the two respective groups were sacrificed at 49 days of age for detecting Toxoplasma gondii by using Species-specific PCR technique. Results of Latex agglutination test indicated, principally, that 29.3% and 49.2% of the serum samples were positive for the birds of both groups, respectively. Titer figures ranged between 1:20 to 1:320 where the highest value was 1:160 (39.3%) and the lowest was 1:20 (5.8%). Confirmation of 38 and 64 serum samples, using Latex agglutination test was performed by PCR technique, from the two respective groups of chickens. Of those, 8 samples from the college birds and 35 from the commercial farm birds were confirmed positive by giving band of 133 bp, according to specific primers designated on gene B1.Based on these results, pursuing the PCR technique is considered, so far, a most sensitive method for Toxoplasma gondii detection. Also, positive PCR results are counted on as an early marker for reactivation and useful means in monitoring therapies.