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

Abstract

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.

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

“…Microscopic examination it's insufficient to detect the low level of Anaplasma infection (18)(19)(20), so that molecular methods used for detecting the undetected cases of infected animals (21,22). Using of the PCR technique for more specific and sensitive detecting of minimum level of Anaplasma in infected animals (23)(24)(25)(26).…”

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

“…Transferring the samples in a cold box to the parasitology lab of the College of Veterinary Medicine, Al-Qadisiyah University, gross examination of the samples grossly for collecting the adults, isolated worms were taken by forceps to diagnose species of parasites, depending on the diagnostic characteristics described by (1,15). Molecular identification of the nematodes uses extracted of 20 rDNA isolates and traditional PCR technique with higher purity of DNA (16)(17)(18), PCR master mix prepared by using AccuPower® PCR PreMix Kit and this master mix done according to company instructions included: DNA template 5µl, Forward primer 1.5µl, Reveres primer 1.5µl and PCR water 12µl. The PCR primers used in this analysis to the amplify ITS2rDNA gene of P. skrjabini, it is being designed according to (9), while COX1mtDNA gene was prepared as (non-degenerate primer) according to the molecular study of other Spirurida (19), these primers were produced from Bioneer company, Korea as following table 1, and reaction conditions of PCR for amplification showed as in the table 2.…”

Molecular characterization and phylogenic analysis of Anaplasma spp. in small ruminants from Sulaymaniyah governorate, Iraq

“…The forward primer 5′-TACCTTGTTACGACTT-3′ and reverse primer 5′-TGATCCTGGCTCAGAACGAACG-3′ were applied for amplification of the 1462 bp portion of the 16S rRNA gene of Anaplasma spp. (15,16).…”