Molecular Detection and Phylogenetic Analysis of Anaplasma Spp. in Cattle in Al-Qadisiyah Province of Iraq

Abstract

Anaplasma spp. are widely spread rickettsial bacteria transmitted by ticks and placing high impacts on veterinary and public health. A limited number of studies have been carried out on Anaplasmosis in the central part of Iraq. This study was conducted to determine the presence of Anaplasma spp. in cattle in Al-Qadisiyah province, Iraq. A total of 400 blood specimens were collected from cattle suffering from heavy tick infestation. Cattle were blood-sampled from four hyper-endemic areas with ticks. Blood samples were screened using microscopic and polymerase chain reaction (PCR) methods. Diff-quick stained blood smears revealed Anaplasma-like inclusion bodies in 254 (63.5%) samples. According to the 16S rRNA-gene-based PCR analysis, Anaplasma spp. was detected in 124 of the 400 (31%) samples, divided as 96/254 (37.8%) among the microscopical positive samples and 28/146 (19.17%) among the microscopical negative samples. Phylogenetic analysis based on the partial 16S rRNA gene sequencing of ten-PCR positive samples were 99–97% identical to sequences deposited in the GenBank, revealing presence of A. phagocytophilum, A. marginale and unnamed Anaplasma spp. in 40%, 20%, and 40% samples, respectively. Relationships among Anaplasma spp. infections and cattle breed, age, and sex were analyzed. Calves less than one year old showed significantly higher rates (p<0.005) than those from other age groups, whereas sex and breed demonstrated no significant differences (p˃0.001). This study shows that a variety of Anaplasma spp., were endemic in central part of Iraq and is still a hidden problem in cattle in the hyperendemic areas of tick, which requires serious control strategies.

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

“…The ticks which transmit a wide range of veterinary important pathogens, are hyperendemic in the middle and south of Iraq (14). In Iraq, Recent studies of A. ovis and A. marginale in Al-Qadisiyah province, Al-Nasiriyah city and Duhok province were detected in significant infection rate in cattle, sheep and goat respectively (15)(16)(17). The most clinical signs which observed by clinical examination on infected sheep were agreed with Ayyz et al (15).…”

“…In Iraq, Recent studies of A. ovis and A. marginale in Al-Qadisiyah province, Al-Nasiriyah city and Duhok province were detected in significant infection rate in cattle, sheep and goat respectively (15)(16)(17). The most clinical signs which observed by clinical examination on infected sheep were agreed with Ayyz et al (15). 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).…”

Application of a nested polymerase chain reaction assay to detect Anaplasma bovis and Anaplasma phagocytophilium in cattle in Lorestan province, Iran

“…The overall prevalence of A. phagocytophilum in cattle in Lorestan was lower than the prevalence reported in cattle in different zones of Iran (15.5%) and Gilan province, north of Iran (24.6%) but higher than those reported in the center(1%) of Iran (Noaman and Moradi, 2019;Noaman et al, 2016;Salehi-Guilandeh et al, 2018). The prevalence rate of A. phagocytophilum in cattle was also reported in Iran's neighbouring countries inclusive of 2.66% in Pakistan (Iqbal et al, 2019), 30.8% in Turkey (Aktas and Özübek, 2015), and 40% in Iraq (Ayyez et al, 2019) as well as other Asian countries, including 2.1% in South Korea (Seo et al, 2018), 2.4% in China (Yan et al, 2020), and 2% in Japan (Ybañez et al, 2013). However, some disagreement among reports could originate from the different locations with various climatic conditions, different seasons, tick generation, various diagnostic methods (with different specificity and sensitivity), and the difference in the number of samples (Yan et al, 2020).…”