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 prescapular 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.

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 their morphological features. Using mitochondrial cytochrome oxidase subunit  (Cox) 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 widespread ticks that affect cattle in Al Najaf province, Iraq.

The present study was performed to detect the molecular and the phylogenetic identification of species that belonging to the genus of Moniezia Blanchard, 1891 which affected intestines of sheep in Al-Diwaniyah city, Iraq; fifty intestine samples were sought for the infestation of Moniezia spp. from the city slaughterhouse from 1 October to 30 November 2017, this tapeworm was found to infest the intestines of 13 sheep. For morphological identify the genus of this tapeworm, eggs from one gravid proglottid of the thirteen worms were examined, polymerase chain reaction (PCR) and the PCR-productbased sequencing were applied on 4 Moniezia tapeworms targeting a specific region of the 18S rRNA gene. The sequencing has shown 2 species of Moniezia, SP1 and SP2 ,these two species revealed close matching on the phylogenetic tree to an according to the current study findings, Moniezia spp. affect on sheep in the city of Al-Diwaniyah, Iraq, these findings give interesting information about the evolution history of this worm in the studied city.

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.

This evolution-based study aimed to reliably identify the epidemiological prevalence of Escherichia coli that was recovered 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.

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 allelegenetic 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.

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.

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