Author : Monyer A. Al-Fatlawi

Microscopic and PCR-based detection of Babesia bovis and Babesia bigemina in camel females from Al-Diwaniyah Province, Iraq

Monyer A. Abdulameir Al-Fatlawi; Ghaidaa Abass; Zainab Hussein chaid

Iraqi Journal of Veterinary Sciences, In Press
DOI: 10.33899/ijvs.2022.133428.2226

This study was carried out to identify the etiological pathogens responsible of protozoal-like disease conditions occurred in camel females from Al-Diwaniyah Province, Iraq. For this reason, 125 camel females (one blood sample per animal) that showed signs of weakness and pale mucus membranes were considered for the study. The samples of blood were explored microscopically and via the use of a polymerase chain reaction (PCR) method that the targeted glutamine-dependent carbamoyl phosphate synthase (CPSII) gene for identifying Babesia bovis and 18S rRNA gene for detecting B. bigemina. The results of the microscopic technique uncovered the occurrence of babesia spp. in 76 (60.8%) of the examined samples, which encourage the use of the PCR to identify the protozoal species. The PCR findings demonstrated that B. bovis and B. bigemina were detected in 8 (8.9%) and 11 (12.22%), respectively, of the positive microscopic samples. The study findings reveal that weakness and paleness of mucus membranes in camel females can be attributed to the presence of infections by blood protozoa, mainly Babesia bovis and Babesia bigemina.

Indicative parameters for liver fascioliasis at pre-clinical and clinical phases in cows from Al-Diwaniyah city, Iraq

Nawras K. Kadhum; Salah M. Karim; Khalefa A. Mansour; Monyer A. Al-Fatlawi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 3, Pages 653-657
DOI: 10.33899/ijvs.2022.132266.2076

The current work emphasized understanding the liver functions while having pre-clinical or clinical fascioliasis accurately detect the disease phase from cows in Al-Diwaniyah City, Al-Qadisiyah Province, Iraq. The experimental design included the use of 30 cows in Al-Diwaniayh province divided into ten clinically healthy cows (control group), ten acutely infected cows with no apparent clinical signs (pre-clinical group), and ten chronically infected cows with observed clinical signs (clinical group), such as yellowish discoloration of the mucus membranes. Blood samples were collected from each cow for performing the following tests: Alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate transaminase (AST), gamma-glutamyl transferase (GGT), serum levels of pyruvate, lactate, and Cholesterol. The results showed that all the enzymes from the pre-clinical group were significantly (P<0.05) higher in their serum levels compared with those from the control group to elevate to significantly (P<0.05) higher levels in the clinical group in comparisons with those from the control and pre-clinical groups, excluding the ALT that revealed no significant (P>0.05) difference between the pre-and clinical groups. A significant (P<0.05) increase was seen in the pre-clinical group compared to those from the control group, with no significant (P>0.05) difference between the pre-and clinical groups. No significant (P>0.05) differences were recorded between all study groups at serum pyruvate and lactate levels. The present study reveals that the fascioliasis, dependent on the infestation phase, can progressively change the serum levels of the parameters mentioned above and may feasibly be used together as indicators for the accurate detection of the disease stage.

COX1 gene and ITS-2 region: A comparative study of molecular diagnosis of Parabronema skrjabini in camels (Camelus dromedaries), Al-Najaf Province, Iraq

Monyer A. Al-Fatlawi; Haider H. A-Fatlawy

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 77-84
DOI: 10.33899/ijvs.2021.129228.1634

The current study was contributed to the analysis of the nucleotide sequence pattern of the nucleotide sequence of the tissue DNA isolates based on the Internal Transcribed Spacer ribosomal DNA (ITS2rDNA) gene and Cytochrome c oxidase subunit 1 mitochondrial DNA (COX1 mtDNA) using the traditional polymerase chain reaction (150 samples of abomasum collected directly from camel carcasses after working in Al-Najaf slaughterhouse from 1/10/2019 to 1/2/2020). ITS2rDNA were well suited for the prepared primer with size783 bp and identical ratio ranged 81.17-99.73% of the same species, as indicated a high similarity of the isolates taken from two-humped camels in China or less related to Parabronema skrjabini in sheep and goat. In addition, the study identified the number of the mutations within the four COX1 gene and ITS-2 region, which were the most conservative region of the host's species, supporting the concept of host specificity with Parabronema skrjabini. The COX1 gene and ITS-2 region applied to confirm the diagnosis using a universal primer, as it included eight isolates with a size of 689 bp, identical values were ranged from 84.99-98.02% depending on the multiple sequence alignment and showed an increase in the substitution level among isolates at an upper taxonomic level. Studying of the COX1 gene and ITS-2 region in Parabronema skrjabini demonstrated a significant relation in the cluster and an early common ancestor with isolates of the two-humped camel (China). As for the COX1 gene and ITS-2 region, the phylogenetic relationship supported the ribosomal gene results, especially with Habronema muscae or related species such as Habronema majus, Dirofilaria repens, Dipetalonema evansi, Setaria tundra, Cercopithifilaria sp towards the root node. Therefore, considering COX1 gene and ITS-2 region as an ideal tool in determining the phylogenetic history of the sequence maps, but less conservative mode than the ITS2 ribosome gene based to a taxonomic species level.

Molecular characterization and phylogenetic analysis of Escherichia coli isolated from milk of cattle affected by mastitis

Azhar A. Neamah; Khilood H. Fahad; Jenan N. Sadeq; Monyer A. Al-Fatlawi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 251-254
DOI: 10.33899/ijvs.2021.129934.1702

This evolution-based study aimed to reliably identify the epidemiological prevalence of Escherichia coli that wasrecovered from affected milk of cattle by mastitis, study the evolution of this bacterium, and describe some isolates using polymerase chain reaction (PCR) technique and DNA sequencing. Here, we collected 50 cattle milk samples and submitted them to conventional bacterial isolation and identification using enrichment culture method and biochemical tests. Then, we confirmed the results by PCR technique based on 16S ribosomal RNA gene. The results showed that E. coli was isolated from cattle at (36%), and this was confirmed by PCR that showed highly specific detection of E. coli isolates at (100%). DNA sequencing of partial 16S ribosomal RNA gene showed (99%) homological identity with NCBI-Blast E. coli isolates and the phylogenetic analysis showed genetic similarity (0.5 genetic changes). In conclusion, this was the first study in Iraq to report genetic relationship between E. coli isolated from milk of mastitis-infected cattle. Therefore, it is essential to define the role of animals as an important source in the distribution of some pathogens that are related to public health.

Morphological and phylogenetic characterization of Oestrus ovis larvae in sheep: Al-Qadisiyah province, Iraq

Nadia S. Alhayali; Mohenned A. Alsaadawi; Monyer A. Al-Fatlawi; Mansoor J. Alkhaled

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 133-137
DOI: 10.33899/ijvs.2021.129529.1656

The fly larvae infect the nasal cavities and sinuses (frontal and maxillary) of sheep, goats, and a range of wild ruminants, forming a disease called oestrosis (Nasal myiasis or nasal bot). The disease is one of the significantly diseases for the Iraqi small ruminant industry that causes detrimental economic losses. The current work was carried out to morphologically- and molecularly-characterize O. ovis larvae collected from sheep in a slaughterhouse in Al-Qadisiyah province, Iraq. The study depended on collecting 20 larvae (at different stages) from 20 sheep from 15 October till 17 December 2020. The morphological examination was done using a stereomicroscope and relying on larval characteristic features, including the posterior end, spiracles, and cephalopharyngeal skeleton. The molecular characterization was performed utilizing polymerase chain reaction (PCR) and partial gene sequencing (PGS) methods of the cytochrome c oxidase subunit I (cox1) gene at 700-bp and 300-bp regions. Morphologically, the first-stage larvae (L1) showed characteristic mouth hooks, while the second-stage larvae (L2) revealed clear terminal stigmas. For the third-stage larvae (L3), the color of body segments and their spines' were the most important features for this larval stage. The PCR showed amplification at both regions 700bp and 300bp, in 8 and 7 isolates, respectively. The PGS revealed 15 different local isolates in genetic level aligned with isolates from Kyrgyzstan, Italy, Spain, and Turkey. This study shows the important strain differences of O. ovis that infect the local sheep in Al-Qadisiyah province, Iraq.

Major-surface-protein-4-gene-based detection of Anaplasma marginale isolated from sheep in Al-Diwaniyah province, Iraq

Saba F. Klaif; Asaad J. Abid; Monyer A. Al-Fatlawi; Mansoor J. Ali

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 85-88
DOI: 10.33899/ijvs.2021.129230.1635

This study was purposed for confirming detection and typing of Anaplasma spp in infected sheep from Al-Diwaniyah province, Iraq. Sampling of 50 blood specimens was performed from clinically-identified infection of anaplasmosis. The samples of the blood were subjected to DNA extraction followed by polymerase-chain-reaction (PCR)-based detection of the Anaplasma marginale using major surface protein (MSP4) gene. The results have shown that 8 blood samples were infected with A. marginale. The PCR-based identification has revealed a confirmative identification of the Anaplasma marginale in the infected sheep. The study identifies Anaplasma marginale as a member of infectious agents that affect sheep in the study city. 

Genotypic analysis of ticks species infesting cattle in Al-Diwaniyah abattoir

Mansoor J. Ali; Wisam R. Raheem Atiyah; Monyer A. Abdulameir Al-Fatlawi; Saba F. Khlaif

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 4, Pages 673-677
DOI: 10.33899/ijvs.2020.127772.1525

Different vectors are considered critical for disease transmission between animals; however, ticks play a significant role in the dissemination of various infectious illnesses of animals and human importance. The current work was carried out to categorize ticks genetically of those isolated from cattle that entered Al-Diwaniyah abattoir. In the present study, 50 tick samples were collected and subjected to microscopic examination and genetic-based methods of polymerase chain reaction and partial gene sequencing, both utilized the mitochondrial cytochrome c oxidase subunit I (COX1) gene as a genotyping element. The findings of the microscopic examination showed that the ticks were from Hyalomma spp. Further analysis, the polymerase chain reaction revealed the genus of Hyalomma of the ticks, but when the PGS was performed, one sample of H. detritum, three samples of H. excavatum, and two samples of H. marginatum were identified. When the phylogenetic analyses were conducted, H. detritum showed close genetic similarity to an isolate from Spain EU827695.1. H. excavatum revealed similarity with isolates from India MK863382.1 and Turkey MT230050.1. In contrast, H. marginatum displayed close identity to an isolate from Iran (MG557555.1). In conclusion, these findings may indicate evolutionary links of the locally identified isolates to different world isolates, probably due to the trade-moving of animals.

Morphological and molecular identification of Parabronema skrjabini of camels (Camelus dromedary) in Najaf province

Haider H. A-Fatlawy; Monyer A. Al-Fatlawi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 3, Pages 507-512
DOI: 10.33899/ijvs.2020.127101.1459

The current study was conducted during the period from September 2019 to December 2019, the number of examined samples 150 abomasums isolated from males 127 and females 23 to identify the species of Parabronema skrjabini that infected one-humped camel slaughtered in Al-Ashraf Najaf slaughterhouse. The microscopic examination of the worms was properties compared to other worms of the digestive system, and then confirmed using genetic markers with the polymerase chain reaction (PCR). Finally, the use of sequencing and phylogenetic analysis technologies relative to those that are predominant in world regions registered in the Gene bank. The results of the microscopic examination showed that Parabronema skrjabini distinguished by a red color, females are curved dorsally and longer than males with a vertically curved and head features that resemble a horseshoe for both sexes. The number of infected samples is 53 (35.33%) and the non-infected samples 97 (64.64%). The highest rate of infection during December month 63.41%. On the other hand, the prevalence rate has been reported 65.21% females and 29.92% males with significant differences. In this research, PCR technique was used the molecular examination with the selection of the highest DNA samples, which were 10 samples to determine the alignment range according to the ITS2 gene, all samples were well suited to primer in length 783 bp and confirmed the diagnosis of these nematodes.

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.

Clinical and molecular identification of ruling Theileria annulata strains in cattle calves in Al-Diwaniyah province, Iraq

Monyer A. Alfatlawi; Asaad A. Jasim; Noor E. Jarad; Saba F. Khlaif

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 115-119
DOI: 10.33899/ijvs.2020.126429.1319

This study aimed to investigate the evolutionary status of T. annulata in Al-Diwaniyah province, Iraq. In this study, the clinical examination of 50 infected animals was performed with blood sample collection (2.5ml per animal), and drug targets cytochrome b, a vital component of the electron transfer chain in the mitochondria of the protozoan, cytb gene was targeted using a polymerase chain reaction (PCR) procedure. Also, 18S rRNA gene as a molecular target for the PCR and a partial gene sequencing (PGS) were included. The PCR that involved using the 18S rRNA and cytb genes as genetic targets revealed amplification of the targeted pieces at 620bp and 1092bp, respectively, in all tested samples. The18S rRNA gene sequence of local T. annulata isolates were aligned with global reference strains for T. annulata recorded in the GenBank. The local strains were close, 100%, in their identity to isolates from Iran, Turkey, and Pakistan; however, they were 99% similar to a nucleotide sequences from India and Bangladesh. Diseased calves showed clinical signs such as high fever (40.3-41.5°C), decreased appetite or in appetence, asymmetrical enlargement of superficial lymph nodes particularly the pre-scapular ones, some cases with diarrhea, pale or icteric mucus membrane of eyes, bulging eyes, lacrimation, ecchymotic hemorrhages on the sclera, incoordination, nervous signs (Dullness, depression, lethargy), salivation, and bloated young calves. The data observed from the present inspecting work may reveal genetic evolution in the local strains with others recorded in the GeneBank. This means that our local strains might have close relationships with some global strains.

Synergizing the deltamethrin larvicidal activity against Aedes albopictus larvae using cinnamaldehyde in Diwaniyah, Iraq

Mansour J. Ali; Azhar C. Karawan; Dhafer R. Al-Fetly; Monyer A. Alfatlawi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 317-320
DOI: 10.33899/ijvs.2019.126026.1212

The current work on mosquito larvae was performed to evaluate the resistance status of larvae to deltamethrin (DM) and to detect if the larvicidal activity (LA) of this chemical could be synergized after exposing the larvae to cinnamaldehyde (CD). Here, 200 Aedes albopictus larvae were employed for the experiment and were divided randomly into 2 groups (100/each group and placed in petri-dishes (PD), 10 larvae/PD), and they are the DM group (1ml of 0.04 mg/l in 99ml of distilled water (DW) was placed to each PD) and the DM+CD group (1ml of 0.04 mg/l and 1ml of 0.9mg/l respectively were placed with 98ml DW in each PD). The experiment was lasted for 24hrs. Larvae were detected to have resistance against DM as 45% to 60% of the larvae were killed by the DM, 40% to 55% resistance rate. However, when evaluating DM activity with the use of CD, the LA was synergized showing mortality in 87% to 92% of the larvae in which a significant increase in the mortality in DM+CD group was noticed more than that in the DM group. Furthermore, RT-qPCR was run to identify the expression status of the P540 monooxygenase gene, Cyp6p15, and found that the gene expression was significantly inhibited in the DM+CD group when comparing that in the DM group that showed overexpression of this gene. This work results provide viable information about the potential activity of the cinnamaldehyde in synergizing the larvicidal activity of deltamethrin.

Molecular identification of allelic genotypes of pyrethroid-insecticide resistance in housefly, Iraq

Monyer Abdulameir Abd Alfatlawi; Mansour Ali; Hassan Naser

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 209-212
DOI: 10.33899/ijvs.2019.125538.1060

According to global-wide presence of insecticides resistance to pyrethroids, the current study identified the purpose to detect the allelic genotypes regarding this issue in house flies in Iraq. From the governorate of Al-Qadisiyah, Iraq, 60 morphologically and molecularly recognized house flies were caught from 6 different regions. Using a technique called polymerase chain reaction (PCR) amplification of specific allele (PASA), PCR was employed to reveal the presence of allele-genetic variations in the para-type sodium channel (para) gene to recognize knockdown resistance (kdr) mutation from the homozygous-wild type of complete susceptibility (sus/sus) to the mutated-homozygous type of complete resistance (kdr/kdr) or to the mutated-heterozygous type (kdr/sus). Here, these genotypes were targeted using specific primers to identify these genetic variations. The results have declared the presence of the sus/sus at 100%-frequency rate in all flies, and none of the other genotypes were detected (0%) in all flies. This valued piece of result indicates the reality of resistance persistence due to lack of insecticide-spraying programs in the governorate. This study provides high-quality information about the current status of insecticide resistance in house flies in Iraq about supporting the fact of genetic-base development of such resistance via frequent use of insecticides.

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.

Morphological and phylogenetic study of Hyalomma anatolicum in Al-Najaf, Iraq

M. A. AL-Fatlawi; M. J. Ali; H. H. Albayati

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 2, Pages 261-266
DOI: 10.33899/ijvs.2019.153860

Studies had been previously conducted to genetically identify species of ticks in Iraq. Therefore, the current investigational study was conducted to recognize the species of 50 ticks collected from infested skin of cattle. The current study defined the ticks to be from Hyalomma genus depending on theirmorphological features. Using mitochondrial cytochrome oxidase subunit I (CoxI) gene, 16 ticks were further confirmed using polymerase chain reaction (PCR). Two PCR products were subjected to DNA sequencing to name the species of the ticks and compare them to some other known ticks in neighbor and world countries. The sequencing results identified the ticks to be Hyalomma anatolicum. One isolate is closely similar to Indian and Iranian isolates, and the other isolate is clustered alone by itself. The results indicated that H. anatolicum is one of the wide-spread ticks that affect cattle in Al Najaf province, Iraq.