Genetic characterization of highly pathogenic avian influenza A (H5N8) virus isolated from domestic geese in Iraq, 2018.

BACKGROUND: Influenza viruses are a continuous threat to avian and mammalian species, causing epidemics and pandemics. After the circulation of H5N1 in 2006, 2015, and 2016 in Iraq, an H5N8 influenza virus emerged in domestic geese in Sulaymaniyah Province, Iraq. This study analyzed the genetic characteristics of the Iraqi H5N8 viruses. RESULTS: An HPAI virus subtype H5N8 was identified from domestic backyard geese in the Kurdistan Region, north Iraq. Phylogenic analyses of the hemagglutinin (HA) and neuraminidase (NA) genes indicated that Iraq H5N8 viruses belonged to clade 2.3.4.4 group B and clustered with isolates from Iran, Israel, and Belgium. Genetic analysis of the HA gene indicated molecular markers for avian-type receptors. Characterization of the NA gene showed that the virus had sensitive molecular markers for antiviral drugs. CONCLUSIONS: This is the first study ever on H5N8 in Iraq, and it is crucial to understand the epidemiology of the viruses in Iraq and the Middle East. The results suggest a possible role of migratory birds in the introduction of HPAI subtype H5N8 into Iraq.

Genetic characterization and phylogenic analysis of H5N1 avian influenza virus detected in peafowl in Kirkuk province, Iraq.

A highly pathogenic avian influenza (HPAI), H5N1, was detected for the first time in peafowls in Kirkuk province, Iraq in 2015. Genetic analysis of the Kirkuk H5N1 indicated molecular markers for avian-type receptors. The Kirkuk H5N1 hemagglutinin gene had an infrequent amino acid cleavage site (SPQREKRRKRGLF), and neuraminidase genes showed sensitive molecular markers for antiviral drugs. Additionally, the phylogenetic analysis found that the Kirkuk H5N1 belonged to subclade 2.3.2.1c. Our results showed that the 2015 H5N1 from the Iraqi city of Kirkuk exhibited new genetic characterization and was different from the 2006 H5N1 isolate from Iraq.

Molecular detection of Theileria spp. in native sheep and estimation of hemato-biochemical parameters from Sulaimani province/Iraq.

Theileriosis, the hemoprotozoan infection, is an endemic condition in tropical and subtropical areas. In this study, conventional PCR analysis was applied to detect the natural infection of native sheep with theileriosis and estimate its effect on hemato-biochemical parameters. The study was carried out in five regions of Sulaimani province, northern Iraq. From May to October 2021, a total of 360 blood samples were collected randomly from the jugular vein of sheep belonging to 23 flocks with a history of tick infestations. After PCR for theileriosis, the hematobiochemical parameters were evaluated by an automatic analyzer using commercial kits. The PCR results represented that 71.7% of the examined sheep were infected with Theileria parasites. The highest prevalence rate (74.6%) was reported in Said Sadiq, and the lowest prevalence (69.5%) was from Bazian. The infection rates in Mawat, Qaradagh, and Sharazoor were 73.1, 70.3, and 71.8%, respectively. The hemogram data revealed a significant decrease in erythrocyte count, hemoglobin concentration, and hematocrit values. Erythrocyte indices also showed significant increases in MC, MCH, and MCHC levels, but no significant differences were detected between the counting of leukocytes, lymphocytes, and granulocytes. Biochemical analysis revealed a significant decrease in total protein, albumin, and creatinine levels with a significant increase in urea and AST levels in infected sheep with theileriosis. Alteration in hemato-biochemical parameters from infected animals can outline the impact of theileriosis on body metabolism and blood factors in naturally infected sheep.

Expression Level of the mip, pmp18D, and ompA Genes in Chlamydia abortus Isolated from Aborted Ewes.

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.

Divergent Analyses of Genetic Relatedness and Evidence-Based Assessment of Therapeutics of Staphylococcus aureus from Semi-intensive Dairy Systems.

Use of antibiotics without following standard guidelines is routine practice in developing countries which is giving rise to genetic divergence and increased drug resistance. The current study analyzed genetic divergence and drug resistance by S. aureus and therapeutic efficacy of novel antibiotic combinations. The study revealed that 42.30% (minimum 20%-maximum 70%) of milk samples are positive for S. aureus. Study also revealed seven SNPs in the S. aureus nuc gene (c.53A>G, c.61A>G, c.73T>C, c.93C>A, c.217C>T, c.280T>C, and c.331T>A). Local isolates Staph-2 and Staph-3 were closely related to Bos taurus nuc gene (bovine S. aureus), while Staph-1 was closely related to Homo sapiens (human S. aureus) indicating shifting of host. Change of two amino acids and staphylococcal nuclease conserved domain was observed in all local isolates of S. aureus. The isoelectric points predicted by protParam of Staph-1, Staph-2, and Staph-3 proteins were 9.30, 9.20, and 9.20, respectively. The antibiotic susceptibility profile of S. aureus presented highest resistance against penicillin (46.67%) and glycopeptide (43.33%). When a single antibiotic regimen was adopted in a field trial, the highest efficacy was reported in the case of oxytetracycline (80%) while lowest was presented by azithromycin. Among antibiotics' combined regimen, the highest efficacy (80%) was presented by gentamicin with oxytetracycline: cefotaxime with vancomycin; and ciprofloxacin with vancomycin. The current study concluded rising percentages of S. aureus from dairy milk, proofs of genetic host shifts, and altered responses of in on field therapeutics.

First Report of Multiple Drug-resistant Gastrointestinal Nematodes of Sheep in Arbat District, Sulaymaniyah, Iraq Detected By in Vivo and in Vitro Methods.

INTRODUCTION: The control of gastrointestinal nematodes (GINs) in small ruminants is principally dependent on anthelmintic therapy, which encounters the rising problem of anthelmintic resistance (AR) development. Veterinarians reported anthelmintic failure in several sheep farms in Arbat District, Sulaymaniyah, northern Iraq, which called for a systematic study about the efficacy of three commonly used drugs: albendazole, ivermectin, and levamisole. MATERIAL AND METHODS: A faecal egg count reduction test (FECRT) was conducted to elucidate the anthelmintics' efficacies, with coprocultures to determine the GINs parasitising sheep in the study area. Larval development assays (LDAs) were conducted to determine the drugs' median inhibitory concentrations (IC50). RESULTS: The FECRT revealed that AR was widespread to all three drugs, and ivermectin was the least effective, reducing the faecal egg counts (FECs) by 50.5% to 57.1%. The coprocultures revealed that the GIN genera of sheep in Arbat District were Trichostrongylus, Nematodirus, and Trichuris, and the resistance was mainly due to Trichostrongylus species. The mean IC50 of albendazole, ivermectin, and levamisole were 0.073 ng/mL, 7.97 ng/mL, and 1.43 ng/mL, respectively. CONCLUSION: This study is the first Iraqi report of AR confirmed by both in vivo FECRT and in vitro LDA methods.

Comparison of a Barium Chloride Test with ELISA for Pregnancy Detection in Cows.

INTRODUCTION: Early detection of pregnancy is vital for appropriate reproductive management programmes to facilitate the rapid re-insemination of non-pregnant females and reduce the calving interval. MATERIAL AND METHODS: A barium chloride test was compared with a commercial progesterone ELISA to detect pregnancy in non-descriptive cows and investigate if it could be applied as an alternative to ELISA in the field. Blood and urine samples were collected from 74 cows with recorded insemination dates. The progesterone ELISA and barium chloride assay were implemented to detect progesterone (P4) in blood and urine specimens, respectively. The cows' reproductive systems were examined after they were slaughtered to determine the uterus's status. Macroscopic examination of the uterus was used as a reference standard for both tests. RESULTS: The sensitivity rates of the P4 ELISA and barium chloride test to detect pregnant cows were 100.0% and 79.4%, and to detect the corpus luteum (CL) were 83.0% and 87.0%, respectively, their sensitivity increasing in the presence of the CL. The ELISA and barium chloride tests were 79.7% and 52.7% accurate in the diagnosis of pregnancy. The accuracy of the barium chloride test in CL detection increased to 81.0%, and that of the ELISA to 86.4%. There were no significant differences (P = 0.052) between the barium chloride assay and ELISA when they were utilised for the identification of the CL. CONCLUSION: The barium chloride test can be an inexpensive and time-saving alternative to ELISA in pregnancy diagnosis when the insemination date is known.