Abortion in small ruminants is a significant problem in Iraq and causes severe economic losses in sheep farms. Chlamydia abortus causes enzootic abortion in ewes and is associated with reproductive problems in sheep in Sulaimani province – Northern Iraq. During a lambing season in 2017, abortion was widespread among several sheep flocks in different regions of Sulaimani (Kalar, Said Sadiq, and Chamchamal), and C. abortus was one of the causes. Accordingly, we carried out this study to isolate and identify C. abortus in aborted ewes in these regions. We collected 30 samples of aborted fetuses from five herds in which abortions had been observed. The pathogen isolation was done by inoculation into embryonated chicken eggs and conventional PCR was used to identify C. abortus in clinical specimens. C. abortus was identified in one of the 30 aborted fetuses (3.33%) from the Kalar district, and all the remaining 29 samples (96.66%) were found positive to Brucella abortus. The gene ompA encoding the outer membrane protein of C. abortus was sequenced and got the accession number MK643153 in NCBI GenBank. The sequence was named C. abortus strain Sul/2017. Our isolate showed 99.79% homology with Sul/014 (accession No. KY399850) and differed from the latter by two amino acid substitutions at E115K and K259N. The topology of the phylogenetic tree based on the ompA gene showed that the isolate belongs to C. abortus and has a common ancestor with isolates of sheep in Iraq and Tunisia with accession numbers KY399850 and HQ62243, respectively.
Abortion is one of the most critical factors affecting lambing rates and, as a result, sheep farm profitability. It is also significant from a zoonotic viewpoint, in addition to financial losses. In sheep flocks, Campylobacter fetus causes infectious infertility, embryonic death, and miscarriages. The study investigated C. fetus from aborted fetuses and vaginal swab samples collected from sheep flocks in the Sulaimani province by the polymerase chain reaction. Thirty-eight aborted fetuses and 70 vaginal swabs were collected from sheep flocks in three districts of Sulaimani province (Kalar, Said Sadiq, and Chamchamal) from March 2018 to June 2019. The pathogen was identified in clinical specimens using conventional PCR. C. fetus was isolated in 16 of 38 aborted fetuses 42.1% and 13 of 70 vaginal swabs from aborted ewes 18.6%. The C. fetus gene 16S rRNA was sequenced and received the accession number MW694741 in NCBI GenBank. Phylogenetic analysis of 16S rRNA gene sequences designated that the C. fetus isolates formed a separate branch displayed the highest similarity and clustered with MN203686.1 and EU773268.1 accessions in a specific clade. A lower degree of affinity of C. fetus was revealed with Campylobacter coli and Campylobacter jejuni.
Out of 150 eggs were collected randomly as (local egg 50 and imported egg 100) from different markets of Sulaimani city, during the period from Three (3%) of imported eggs were found contaminated with Salmonella. Three different Salmonella serotypes were identified including; Salmonella typhimurium (one strain), Salmonella ohio (one strain) and Salmonella enteritidis (one strain) which represented 1% for each.
In this manuscript, we report the proteins macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) that are expressed in different times of pregnancy in mice infected with Chlamydia abortus. Enzootic abortion of ewes (EAE) by C. abortus, an obligate intracellular pathogen, is a critical zoonotic disease-causing significant economic loss to livestock farming globally. This study was carried out for the detection and characterization of macrophage infectivity potentiator (mip, CAB080), major outer membrane protein (momp, CAB048), and polymorphic outer membrane protein (pmp18D, CAB776) using RT-qPCR. These proteins are believed to be expressed as virulence factors in C. abortus isolated from aborted ewes. BALB/c mice (pregnant and nonpregnant) were used as an animal model to be injected intraperitoneally with C. abortus culture in Vero cells since the endometrial lymphoid tissues of these animals resembles that of ewes. Also, the short duration of pregnancy in mice makes them a suitable animal model for obstetric studies. Tissue samples were taken from the mice after 10, 15, and 20 days of pregnancy to compare the expression of the genes mip, pmp18D, and ompA. Transcription level was quantified using RT-qPCR, the GAPDH transcription quantification, as a normalization signal. Abortion occurred in pregnant mice, and apparent differences between the transcriptional levels of the mip, pmp18D, and ompA genes in the samples taken during different time intervals of pregnancy were not observed (p > 0.05). The result indicated that the three bacterial genes, mip, pmp18D, and ompA, play a role as virulence factors in abortion and are differentially expressed in pregnant and nonpregnant animals. Inactivation of the genes is suggested to confirm the hypothesis.
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