Iraqi Journal of Veterinary Sciences

This research work was conducted as well as the determination of the resistance to antibiotics of these isolated species of E. coli in Nineveh governorate to assess the incidence of Escherichia coli (E. coli) contamination in different fish farms and local fish markets. The total number of fish samples used in the present study was 153, including 75 samples from various fish farms and 78 samples from different local markets in Mosul. The current study showed that the percentage of E. coli isolated from fish farms was 24% (18/75) and 35.9% (28/78) from local markets. While it showed a positive result for E. coli with serotype (O157:H7) with a percentage was 9.3 and 14.1% from both farmed fish and market fish samples, respectively. Additionally, all E. coli positive isolates possess the specific uidA gene, which was detected using the PCR technique. The highest sensitivity of E. coli bacteria to the antibiotic’s ciprofloxacin, trimethoprim, and gentamicin was 96, 94, and 86%, respectively. At the same time, the highest percentage of resistance of E. coli to the antibiotics cephalothin, tetracycline, erythromycin, and amoxicillin was 100, 64, 64, and 62%, respectively. To reduce health risks to consumers, these results provide useful basic information for the proper management of these environments in order to prevent fecal contamination in fish farms and the fish sold in local markets.

This study aimed to characterize and identify the genotypes of Fasciola spp. isolated from sheep, goats, and cattle in Duhok province based on the ITS1 region of rDNA. About 54 adult Fasciola flukes were individually isolated from the livers of naturally infected ruminants. After morphological identification, the genomic DNA of 54 isolated Fasciola spp. was successfully extracted, and the ITS1segment (518 bp) of rDNA was amplified. The amplicons were confirmed by gel electrophoresis and yielded mono cleared bands. Five amplicons from these samples (2 sheep, 2 cattle, and 1 goat) were selected for sequencing and then compared with NCBI-GenBank sequences for genotyping and phylogenetic analysis. Sequencing analysis and the BLAST results revealed that 3/5 of the resultant sequences were F. hepatica and 2/5 were F. gigantica. The ITS1 sequences were submitted to NCBI-GenBank with accession numbers: OM920533, OM920534, OM948733, OM948683, and OM918714. Alignment analysis of the current study and GenBank ITS1 sequences showed the presence of nucleotide variations between F. hepatica and F. gigantica species (interspecific), which were enough to separate them. At the same time, they were not observed within the same species of Fasciola (intraspecific). The pairwise identity percentage of intraspecific and interspecific Fasciola isolates was 100% and 99.2-99.6%, respectively. Phylogenetic analysis of the ITS1 sequences demonstrated that the Fasciola isolates of this study were clustered into two clades (hepatica and gigantica clades). The present study concluded that both Fasciola spp. (F. hepatica and F. gigantica) existed among the infected ruminants in Duhok province and are closely related to intraspecific Fasciola isolates from different countries in the Middle East, Asia, Europe, and Africa.

The aim of this study is to detect the 28S (rRNA) gene sequences of Oestrus ovis larvae by conventional polymerase chain reaction and to compare their genetic relatedness utilizing phylogenetic analysis. Fifty larvae were collected from sheep for DNA extraction after slaughtering during the period from the beginning of February until the end of April 2019 in Baghdad city. PCR product appeared as the band size 950 bp. Ten of the product PCR were selected for sequence analysis to obtain the partial nucleotides 28S (rRNA) gene. After that the sequence were recorded in National Center for Biotechnology Information (NCBI) with ID NO. (MT875427, MT875428, MT875429, MT875430, MT875431, MT875432, MT875433, MT875434, MT875435, MT875436) for O. ovis larvae. Then, compared these accession number with another global registered in NCBI by using phylogenetic tree examination which show NCBI-BLAST homology sequence identity between them, and these results were confirmed 99% identity with Spain and Brazil isolates and 98% with Italy.

Sarcoptes scabiei var cameli is the most frequent zoonotic species of mites causing mange in camels worldwide. The prevalence of camel’s mange in Iraq is still little studied. Thus, this research is conducted to detect S. scabieiin camels in the four provinces of the Middle-Euphrates area: Al-Muthanna, Al-Diwaniyah, Najaf, and Babil, from January 2020 to December 2020. The Molecular technique depending on the conventional polymerase chain reaction (cPCR) is performed for the direct detection of S. scabiei based on the mitochondrial cytochrome oxidase subunit 1 (COX1) gene from skin scrape lesion samples. The results reveal that 125 out of 425 samples (29.41%) of the examined camels are infested with S. scabiei. According to the sex of the infested animals, the infestation rate was higher in females than in males, 85 (30.91 %) and 40 (26.67%) respectively. In addition, the 1.5 year age shows the highest number of infestation (83 out of 85) with a percentage of 97.65%, but the percentages are 21 out of 60 (35%) and four out 68 (5.88%) in 2 and 7 years old animals, respectively. The results also record that infested animals found in Najaf and Al-Diwaniyah have the highest number of infestations, with of 36% and 35%, respectively. The findings also demonstrate that the highest infestation percentage is during the winter months (January and February), with of 92.31% and 80%, respectively. The sequencing and phylogenetic analysis shows that the local isolates of the Iraqi camels are consistent with the isolates recorded in China.

The genetic relatedness of Salmonella enterica sub sp. enterica isolated from human and animal origin has more interest as its data possibly will offer an essential confirmation for the source of human infection. This study aimed to determine the genetic relationship of S. enterica subspecies enterica isolated from human and animal sources. A total of 300 samples were collected from two primary sources, human and animal, from two different regions, Baghdad and Basrah governorates. For constructing the phylogenetic Tree of Salmonella enterica subspecies enterica, the sequencing of PCR product (positive samples) for each target genes 16s rRNA, avrA, and spvC were analyzed using BLAST analysis to determine the similarities and differences between the Iraqi strains and the existing global strains. The similarity rate in the first gene 97.77%, the second gene 98.29%, and the third 96.82%, respectively. The genetic Tree of each of the three genes was set up separately using two methods, Maximum Likelihood, and the second Minimum evolution. The phylogenetic analysis reveals that Iraqi strains of Salmonella are highly similar, and they share the same sequence of 16s rRNA gene with national Salmonella strains. However, their bases of avrA and spvC genes are not similar. This difference leads us to conclude that the Iraqi Salmonella evolution path was characterized by its path in developing global strains with some correlation in some samples; it may be linked with the same ancestors from which it emerged.

Pseudomonas has been recognized as a unique meat spoiling organism. The proliferation of these spoilage organisms might influence the organoleptic meat quality. Therefore, the current investigation is being carried out to detect pseudomonas associated with meat displayed in Mosul city retails. A total of 150 meat samples of beef, mutton and chicken meat (50 of each) were collected. Molecular identification of pseudomonas aeruginosa in meat is performed by targeting the16S rRNA gene and rpoB gene. Fifty-three isolates of pseudomonas species were obtained from all types of meat (35.33%), including 23 (46 %) for beef meat,11 (22%) for mutton and 19 (38%) for chicken meat. Enumeration of pseudomonas species in beef and mutton were 1.47*10⁴, 1.92*10⁴ CFU/g, respectively^, while counts were 21.3*10⁴ CFU/g in chicken meat. Polymerase chain reaction results revealed the presence of 16SrRNA gene in all tested isolates 53/53 (100%). pseudomonas aeruginosawas isolated at (39.62%) from meat samples according to the detection of the rpoB gene.In conclusion, the prevalence of pseudomonas in meat at Mosul city retails negatively impacted meat quality and consumer confidence. Also, the PCR approach aids the rapid detection of pseudomonas as spoilage organisms in meat to reduce financial loss.Therefore, hygienic measurements should be applied to reduce meat spoilage and conserve consumer health during meat production and preservation.

There are 13 virulence-related genes in E. coli isolates. The10 genes of these isolates were selected from avian pathogenic E. coli in some Iraqi broiler farms. Six of these virulence-related genes (iroN, iucC, frz operon, iucD, papC, and R4) were investigated in these isolates by PCR. Eighty percent of the isolates had one or more virulence-associated genes. Two APEC separates carried just one gene, iroN or iucC. According to preliminary evidence, the iroN and iucC genes may express their pathogenicity independently. All of the strains had the same iroN gene, making them all pathogenic. The results of these isolates were confirmed by PCR to have the six pathogenic genes: 80% positive for iucC, 50% positive for iucD, 100% positive for iroN, 10% positive for frz operon, 10% positive for papC, and 0% positive for R4 respectively. These six virulence genes were detected with different percentages in isolates; the iroN gene was found in all isolates but the other virulence genes were found with different percentages in E. coli isolates.According to, detection the iroN and such genes are displaying their pathogenicity separately from each other.

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 rRNAwas 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.

The aim of this study was to provide field outbreaks data with FAdVs in Ninevah governorate to emphasize the importance of the disease due to high mortality and production losses. A total of 729,500 broilers collected from 64 flocks at 14 different locations in Nineveh governorate during the second half of 2020. were included in this study. Histopathological changes of the liver in infected birds have been studied. Molecular identification of FAdV was accomplished by DNA extraction from liver samples using DNeasy Tissue Kit. Results reviled that there were 51892 mortalities representing 7.11%. It was noted that the broiler flocks were infected during their 2^nd-6^th weeks of age, being the highest in the 5^th week of age. Decreased mortality was detected from July to December. being 11.3, 7.91, 7.08, 6.38, 5.94 and 4.95%, respectively. Microscopical examination of the liver manifested the pathognomonic presence of eosinophilic intranuclear inclusion bodies related to the disease. PCR findings revealed positive results of FAdVs. It could be concluded that the environmental stress and immunosuppressive agents could contribute to the percentage and duration of mortalities in broiler flocks. 

The current study examined 100 small intestines collected randomly from sheep slaughtered in the abattoir and butcher’s shops from different Mosul city / Iraq areas of both sexes (55 females, 45 males) and different ages. Moniezia expansa was diagnosed in 9 samples of intestines by studying the morphometric characteristics of these tapeworms, especially the mature segments, in which both the ovaries and vitelline glands appeared in the ring shape on either side of the body segments and the rosette-like shape of the interproglotidial glands. No significant difference was noticed between males and females of sheep in our study, and the infection rate was 10% in sheep less than a year old and older than two years, with no significant difference between the age groups. The results of the molecular analysis by using conventional polymerase chain reaction technique confirmed the diagnosis of these worms, which belong to the genus Moniezia, with a product reaction of 700 base pairs. The sequencing result shows two strains of Moniezia expanza, which isolated from Iraq (Moniezia expansa-Iraqi one and Moniezia expansa-Iraqi 2) were similar to each other had a significant distance to other strains. The study also showed that Moniezia expansa is different from the same species in other countries.

Globally, extended-spectrum ß-lactamase (ESBL)/Ampicillin ß-lactamase (AmpC) producing Escherichia coli has become the greatest threat for distributing antibiotic resistance. Accordingly, this study was designed to detect and screen the genes that confer resistance in E. coli isolated from sheep as main livestock in Mosul city. Forty E. coli isolates previously recovered from milk and fecal samples were included in this study. These isolates were obtained from healthy ewes, their lambs, and also from ewes with clinical mastitis. Polymerase chain reaction (PCR) was used to confirm the E. coli isolates targeting the 16sRNA gene. Furthermore, screening of different genotypes of ESBL/AmpC was conducted using specific primers. The results showed that the CTX-M gene was predominant among ESBL genotypes and recorded 40/40 (100%). While, SHV and TEM genes recorded 7/40 (17.5%) and 5/40 (12.5%), respectively. Moreover, fecal carriage of resistance genes was more than that obtained from milk in both healthy and diseased animals. However, none of the 40 isolates showed positive results for AmpC genes. The presence of different genotypes of ESBL E. coli isolated from feces or milk origin may act as a potential source for transferring antibiotic resistance to humans, other animals, and the environment.

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

Anaplasma spp. are significant arthropod-borne bacteria globally, but documented information about anaplasmosis in small ruminants in the north of Iraq is insufficient. Hence, this study was conducted to determine the prevalence of Anaplasma spp. and identify sheep and goat tick vector populations in Sulaymaniyah Governorate, north Iraq. The study population consisted of 470 sheep and 145 goats from 45 livestock farms in 10 geographical locations of Sulaymaniyah Governorate. The study was accomplished from April to December 2017. Blood samples were taken from the jugular vein and used for DNA extraction. Polymerase chain reaction (PCR) was conducted using primers based on the 16S rRNA of Anaplasma spp. Fragments of PCR products were sequenced for phylogenetic analysis. The prevalence of Anaplasma spp. was 58.9% based on the PCR results. Furthermore, 58.9% of sheep and 57.9% of goats were positive for anaplasmosis. The sequences represented 100% identity with previously documented GenBank isolates of A. ovis from Iran, the Netherlands, China, and Mongolia. Altogether, 150 Ixodid ticks were picked from small ruminants within the same flocks and were identified based on morphological features. Various infestation rates were observed; about 40% of the Ixodid ticks belonged to Rhipicephalus sanguineus, 34% belonged to Rhipicephalus turanicus, 18% were Hyalomma anatolicum, and 8% were Boophilus microplus (Rhipicephalus microplus). The present report is the first molecular study of Anaplasma species in small ruminants from Sulaymaniyah Governorate in northern Iraq to the best of our knowledge. The study concluded that anaplasmosis was endemic in small ruminants from the investigated areas. 

The objective of this study was to develop Polymerase Chain Reaction (PCR) procedure for detection of Salmonella Typhimurium in artificially contaminated chicken meat. The experiments were conducted with various dilutions of Salmonella Typhimurium reference the American Type Culture Collection ATCC (ATCC13311^TM 4.4*10⁷) High concentration 4.4*10³ Colony Forming Units (CFU)/ml, low concentration 4.4*10² CFU/ml, very low concentration 4.4*10¹ CFU/ml inoculated in chicken meat, in order to determine limits of detection (LOD), optimum incubation times 18 to 20 hours of pre-enrichment in Buffered Peptone Water (BPW 1%). Hence, cultural methods and DNA extraction were performed according to kits instruction. The microbiological cultural test was capable to detect 1.76 CFU/mL, whereas PCR examination was able to detect 0.18 CFU/ml of initial dilution of Salmonella Typhimurium inoculated in chicken meat. Interestingly, the results were achieved in a less time period than that of classical culture. The PCR technique is beneficial in the methodology for detection of Salmonella in chicken meat.

 Sarcosystis spp., has a close relationship with muscles due to its unique localization within skeletal muscle in humans and the animals it infects, as the chronic condition of the disease causes significant economic losses, especially in terms of meat production as a result of the formation of cysts, whether macroscopic or microscopic, in their muscle fibers. Sarcosystis tenella and Sarcosystis arieticanis are the most important pathogenic cysts forming in sheep. In this study, 50 samples of diaphragm muscles of sheep slaughtered in the butchers' shops and the Mosul abattoir were examined grossly, histologically, and using PCR technique as a diagnostic tool to identify or diagnose the causative and responsible species of these changes. The diaphragm samples appeared white and pale on the macroscopic examination, while the tissue lesions were characterized by the presence of Sarcosystis in different numbers and sizes among the muscle fibers, which led to the occurrence of zinker necrosis and intense infiltration of inflammatory cells, especially eosinophil, monocyte, macrophage and giant cells, and also oedema and proliferation of fibroblast. With the formation of fibrous tissue whose intensity was inferred (mild, medium and intense) by using the masson’s trichrome stain. The results of the molecular analysis using the nested PCR technique indicated that these diagnosed microscopic cysts belong to Sarcosystis tenella with a reaction product of 800bp and 500bp.

Staphylococcus (S.) aureus is the main facultative organism of contagious intramammary infections from lactating animals. It is considered a major foodborne organism that can cause food poisoning conditions around the world. Camels are very important to the lifestyle of many countries because they can produce milk that contains the major components such as proteins, energy, vitamins, and minerals. The present study used a polymerase chain reaction (PCR) method on a base of the nuc gene as a target gene, which is a specific gene that recognizes the S. aureus amongst other microorganisms. Fifty milk samples have been collected from camels from different areas of the Nineveh Governorate, Iraq. According to the phenotypic characteristics, isolation and identification of S. aureus have been accomplished by characterizing the shape of the colonies, painting the suspected isolates by gram stain, using the biochemical tests such as coagulase and catalase. In this study, S. aureus was isolated from 70% (35/50) camel milk samples. The classical method of identifying the S. aureus isolated from camel milk was consistent with the PCR method. The PCR technique indicated that all positive S. aureus possessed the nuc gene. The increased percentage of S. aureus isolated from the camel milk has a relationship with the type of farm management, poor nutrition, and/or environmental conditions, rather than treatment of the infected camel. The PCR method is considered one of the best-used techniques to identify the S. aureus isolated from camel milk by detection of nuc gene, the specific gene of S. aureus.

This study aimed to detect Brucella antibodies in the sera of dairy cows and to identify Brucella species in the milk of seropositive cows. A total of 100 sera and 100 milk samples were collected from two 50-cows groups (group 1 with and group 2 without a history of reproductive problems and/or decreased milk production). Rose Bengal plate test and indirect ELISA were used to explore Brucella antibodies in the serum samples and thereafter milk samples of seropositive cows were undergone PCR analysis using Brucella genus specific primers and 3 pairs of species specific primers for identification of B. abortus, B. melitensis and B. suis. The RBPT showed 22 cows were carriers for the Brucella antibodies, 18 in group 1 and 4 in group 2 whereas the iELISA showed only 10 cows out of these 22 cows were positive, 9 in group 1 and only 1 cow in group 2. The PCR assay, which was performed on milk samples of the RBPT positive cows, revealed 18 samples were positive for the Brucella genus and the Brucella abortus species and were negative for Brucella melitensis and Brucella suis species. As a conclusion, the results of this study showed that brucellosis has been encountered in cows with or without a history of reproductive problems, and the RBPT followed by PCR assay for milk samples of the seropositive cows could provide more specific detection than performing either test alone and could be more useful for rapid screening of brucellosis in dairy cows.

Shepherd dogs have been implemented in the transmission and distribution of many threatening pathogens. The presence of extended-spectrum-cephalosporin resistant Escherichia coli (ESCR E. coli) in dog feces can constitute a significance risk to human health due to transmission of antibiotics resistance from dogs to humans, other animals and the surrounding environment. Therefore, in this study, phenotypic and molecular characterization of fecal ESCR E. coli were investigated in shepherd dogs accompanied sheepherders in urban areas. Sixty-seven fresh fecal samples were collected from shepherd dogs from different regions of Mosul city. Bacteriological examination of ESCR E. coli was done using MacConkey agar with cefotaxime followed by subsequent PCR confirmation of the CTX-M gene using specific primers and molecular characterization using specific primers directed to CTX-M-1, 2 and 9 groups. The results of bacterial examination showed successful confirmation of ESCR E. coli which has been isolated from fecal samples of shepherd dogs 58.2% (39/67). In addition, detection of CTX-M gene was confirmed in 53.7% (36/67) of E. coli isolates. Furthermore, molecular characterization of CTX-M gene revealed the presence of only one genotype belongs to CTX-M-1. However, both of CTX-M-2 and CTX-M-9 genotypes were not detected in this study. This study concluded that shepherd dogs have an essential role in carrying and spreading of ESCR E. coli especially in urban regions.

Extended spectrum β-lactamases producing Enterobacteriaceae (ESBL-E) have emerged recently as the main cause that facilitates the spreading of antibiotic resistance worldwide. Due to its composition and nutritive values, raw cow milk is vulnerable to bacterial contamination from different sources, especially ESBL-E. Accordingly, present study aimed to detect the ESBL-E in the raw milk of healthy cows. 80 raw cow milk samples were collected from unorganized farms and cows belong to individual owners and investigated for the presence of ESBL-E with the main focusing on CTX-M type. The bacterial isolation was performed using selective MacConkey agar plus cefotaxime (MC⁺), while PCR was used to confirm the species of the isolated bacteria and presence of CTX-M gene. The results showed that 28.75%(23/80) samples were ESBL-E positive and distributed as following, 82.61%(19/23) were pure E. coli isolates, 4.35%(1/23) was pure K. pneumoniae isolate and finally, 13.04%(3/23) were mixed of both E. coli and K. pneumoniae isolates. Moreover, the total number of positive ESBL-E was 26 isolates with the majority of them were belong to E. coli and recorded 84.61%(22/26), while K. pneumoniae was recorded less 15.39%(4/26). Additionally, the CTX-M gene was successfully identified in all ESBL-E positive isolates by using PCR, including E. coli and K. pneumoniae isolates. The results of this study assert the importance of raw cow milk as a potential source of ESBL-E that might be transmitted to humans.

Because there was no such study done on identification of tick species by PCR technique in in Duhok Governorate, therefore present study was done to identify tick species by using molecular study by using of 16S rRNA and DNA sequencing. About 1000 ticks were collected from both sheep and goat, form Duhok Governorate including: Barwaria, Zakho, Sumeil, Mangeshik, Sersing, Shekhan and Akre, between May and June 2016, between April and June 2017. The result found during this study were six species under two genera of the hard ticks were identified by molecular study and sequencing including: three species were under the genus Hyalomma and three species were under the genus Rhipiciphalus that infect small ruminants in Duhok governorate from these species a new species under the Hylomma genra (Hyalomma asiatium asiaticum) with accession number (MN594484), was first time reported in Duhok governorate. Also phylogenetic tree was constructed depend on the 16S rRNA.

This study was conducted to investigate Anoplocephalidia Cestoda in sheep and goat and evaluate the 18s rRNA to genetically differentiate the genera of this family. Sixty sample tapeworms were collected from small intestines of 30 sheep and 30 goats from different slaughterhouses in Al-Najaf and Al-Qadisiyah provinces, during September, 2016 to February, 2017. Based on polymerase chain reaction (PCR) and 18s rRNA gene partial sequencing (18sGPS) methods used, tapeworm infection of sheep and goat’s intestines was 32.9% and 31.4%, respectively. The partial gene sequencing of the 18S rRNA gene showed two closely related isolates of M. benedeni which are aligned distinctly to an NCBI isolate of the same species from China. For T. giardia, the outcomes of the phylogenetic analysis unveiled three distinct local isolates which were similar to an NCBI database isolate from China. The current data ensure the importance of the molecular techniques in differentiating between Thysaniezia (Helictometra) giardi and Moniezia species that were identified for their presence in the small intestines of sheep and goats.

The study was done for described genotypically characterize of Staph. aureus isolated from the oral cavity of canary birds in Mosul city using polymerase chain reaction technique which was achieved by amplifying of the thermonuclear nuc gene specialized with Staph. aureus. Sixty birds were examined from variable ages of both sexes from different regions of Mosul city for the period of 1/5/2018-1/6/2019 was carried out. The results indicate that 35 samples gave Staph. aureus with the percentage of 58.4%. These isolates are positive for pigmentation of mannitol salt agar, hemolysis on blood agar, catalase and coagulase-positive, gram staining and oxidase negative. PCR technique indicate that all 35 isolates were positive for the nuc gene and produce amplicon of 166 bp. These results considered positive and it is very specific for bacterial isolates of staph aureus as well as may be used for strain isolation, characterization, and differentiation from other types of bacteria.

This study were conducted in Al-Diwaniya province, in south Iraq during the period from Februaryto July 2019 to determine the rate of infection of Cryptosporidium parvum in domestic chicken, study the effect of some epidemiological factors such as sex and months on the rate of infection, addition to the molecular identification of Cryptococcus parvum by amplification HSP70 gene by conventional PCR. Number of collected fecal sample was 210 from domestic chicken and stained by Ziehl-Neelsen stain. The results of the microscopic examination showed that 108(51.4%) out of 210 fecal samples were infected with Cryptosporidium spp. The statistical analysis founded no marked difference in prevalence of infection between sexes. Significant difference was recorded between infection rate during the months of the study and higher prevalence of infection rate was observed in March 11.9%, while lowest infection rate was observed in July 5.23%. and June 5.23%. Genomic DNA was extracted from 108 fecal samples and HSP70 gene for C. parvum was amplified by PCR. PCR technique is showed that out of 108 fecal samples 21.3% were positive for Cryptosporidium parvum.

Toxoplasmosis is a cosmopolitan zoonotic parasitic disease of mammals and birds; human infection occurs through consumption of raw or undercooked meat. Little was known about the infection rate of T. gondii among free range local chickens (Gallus domesticus) in Duhok province. Therefore, the present study was carried out to determine the infection rate in Duhok province by using ELISA (IgG) and conventional PCR. A total of 368 blood samples were collected from free range local chickens distributed in five different areas of Duhok province during the period from November 2016 to March 2017. The collected blood samples were from different sexes (hens and cocks) and from different age groups (less than 6 months and older than 6 months). The data found that the total infection rate was (84 / 368) 22.8% by ELSIA. The presence of the infection was confirmed by PCR and DNA sequencing. In this study, there were differences from area to area in the infection rates, the highest rate was reported in Semel district at 33.7% which was significantly (p

A few reports are available for detection of L. monocytogenes in fish in Iraq, however, the current study was undertaken to investigate the potential role of Listeria spp. in common carp fish in Baghdad province, Iraq. A total of fresh thirty raw common carp (Cyprinus carpio) were purchased from fish sellers of various local markets in Baghdad city from (December 2017 to March 2018) The viscera was removed aseptically, the bacterial isolation and identification was conducted by a conventional culture method using Listeria selective media, biochemical tests and Vitek 2 for gram-positive. Pathogenicity of isolates was studied in vivo by inoculating mice with bacterium. Targeting virulence associated genes was used to detect the virulence and to confirm the L. monocytogenes isolates. The isolates were tested for antimicrobial susceptibility by disk diffusion method for 12 antibiotics. The results revealed that 6.66% of L. monocytogenes were identified from common carp fish viscera and the isolates were pathogenic in mice. L. monocytogenes virulence associated genes were detected in both isolates, while L. innocua virulence associated gene (Lin0372) was detected in one of the two isolates. The isolates were resistant to 7 out of 12 antibacterial drugs including tetracycline, ampicillin, methicillin, cefixime, oxacillin, cefotaxime and penicillin G. The results suggest that presence of L. monocytogenes in fish may have a serious role in public health hygienic in humans.

Toxoplasmosis is very important zoonotic disease in the world cause by an obligate intracellular protozoan parasite called Toxoplasma gondii can infect human and all warm-blood animals, beef consider from most important source for infection with T. gondii and there is no really data and study about the rate of the infection in beef in Al-Diwaniyah province, so for this reason the aim of this work was designed. A total of 300 samples which collected from heart, tongue, muscles, of 100 slaughtered beef of local and imported cattle, throughout the period from September 2017 to May 2018, initially examined microscopically for searching on T. gondii bradyzoites then all suspected samples was subjected to conventional PCR technique through B1gene amplification to confirm the infection, in addition to analyzed the recorded data for each sample to determine the effect of some factors on prevalence of infection like organ, season and animal age. Out of 300 tested samples only 53 were confirm positive T. gondii DNA. The infection in local beef was higher (22%) than in imported (13.5%), while there is no difference in infection among different examined organs. Regarding to effect of some factors, the autumn season recorded highest rate of infection with significant differences rather than others seasons in both local and imported beef, whereas, age appears with no effect on infection. The local cattle meat is riskier than the imported due to the higher rate of infection with T. gondii, and the animal age cannot affect on the infection rate, in comparing with the season which play role in this rate.

Paratuberculosis or Johne’s disease is a chronic debilitating disease mainly infects ruminants and caused by Mycobacterium paratuberculosis. Previous serological studies in Mosul city confirm the presence of positive reactants for paratuberculosis in cattle. However, culture methods to confirm the disease need a long incubation period and also special media. Raw cow’s milk is considered as potential source for transmission of M. paratuberculosis in cows’ herds. Accordingly, this study aimed to detect the presence of M. paratuberculosis specifically in the raw cow’s milk using polymerase chain reaction (PCR) technique as a rapid, sensitive and reliable method. A total of 50 samples of raw cow’s milk were collected from cows suffering from emaciation and unresponsive to antibiotic treatment. All the samples were subjected to DNA extraction and direct amplification PCR. The results showed that 3 (6%) out of 50 milk samples were positive for M. paratuberculosis. This is the first study in Mosul city that confirms the presence of M. paratuberculosis in raw cow’s milk using PCR technique. In conclusion, raw cow’s milk could be an important source for M. paratuberculosis infection in dairy cows, and also PCR technique could be helpful in rapid diagnosis of paratuberculosis.

Shigella is an intracellular bacterium can infect both human and animal. Its species especially Shigella dysenteriae cause shigellosis worldwide, with 165 million cases of severe bloody diarrhea and mucoid feces. The aim of this study was to find a rapid, sensitive and specific method for screening Shigella in raw bovine contaminated milk. For this goal, 70 samples of milk collected in sterile containers for isolating of Shigella and culturing it on selective media to identify and characterize its morphology, biochemical and molecular characteristics. This study was compared between three different DNA extraction techniques for polymerase chain reaction (direct DNA extraction using a kit, alkaline DNA extraction, and filtrated milk). Our results showed that PCR was able to detect Shigella in 15 out of 15 cases after the milk samples filtered. In other words, the filter technique can be used to detect Shigella in contaminated milk.

Laboratory methods are essential for the diagnosis of Mycoplasmal infection. There are three laboratory approaches are essential for the diagnosis of Mycoplasmal infection in chicken including direct methods by culture method and polymerase chain reaction, and indirect methods by detection of Mycoplasmal antibodies by serological tests. This study aimed to detection of Mycoplasma by culture and PCR technique. Two hundred seventy-six samples were collected from infected adult boiler chicken in Salah Al-din province which suffering from respiratory signs and /or joint infection, 202 respiratory and 74 articular samples. According to the results of culture, Mycoplasma isolated in rate of 35.1% (36.6% from respiratory samples and 31.1% from articular samples). The sensitivity of culture was 100%, while the specificity of culture was 97.9% when comparing with PCR results. The current study concluded that the respiratory infection was more than articular infections, and Mycoplasma gallisepticum more distributed than Mycoplasma synoviae among chickens.

Toxocara (T.) canis is a nematode parasite of canines; belong to the Ascarididae family, which accidentally infected humans. Puppies expel the eggs with the feces from the fourth week of the life cycle. This study is the first study in Iraq for detection seroprevalence in stray dogs and extended from January to September 2017. Our study was aimed to investigate the seroprevalence of T. canis infection in stray dogs from different areas in the Al-Diwaniya province, Iraqto detection of specific IgG antibodies to T. canis compared to Conventional PCR technique with the effect of the risk factor. One hundred of the blood sample and one hundred of a faecal sample of same dogs after shooting were studied usingindirect ELISA test and PCR. The result revealed that 71% of the dogs had a seropositive result for this parasite by ELISA test. Dog age is an important factor and affects seroprevalence, were shown that positive rate in adult dogs was more 83.05% than the young dogs 53.65%, while no significant between dogs according to sex. PCR technique showed 58% of dogs were positive forinternal transcribed spacer 1 (ITS1) ribosomal RNA. The sensitivity and specificity of ELISA test was 79 and 40% respectively.

Babesia is one of hemoprotozoan parasite transmitted by arthropod vectors which responsible for causing of Babesiosis disease in bovine worldwide. The present study was designed for microscopic identification, molecular, and phylogenetic analysis of Babesia species in buffalo from slaughter house in Al-Najaf city of Iraq. The study performed in three months of summer season (August into September 2017) and animals ages and sex were included in this study. The direct microscopic prevalence results were show highest prevalence of haemoprotozoa prevalence at Babesia sp. 45.74%. The prevalence of Babesia sp. related to animal sex, were show in male 43.48% and female was 52%, with non-significant differences. The Prevalence of Babesia sp. related to age were show 12.50%, 92.86% and 30% in young, adult and old age respectively with significant differences (P<0.05). The prevalence of Babesia sp. related to month of study were show. 28.57%, 62.50% and 42.86 in August, September and October respectively and with non-significant differences. Molecular study results were based on PCR and DNA sequencing method by phylogenetic tree analysis (MEGA 6.0) and NCBI-BLAST Homology Sequence Identity to differentiation Babesia species typing. The Babesia species prevalence results were show identified two Babesia species, high prevalence of Babesia bovis (38.30%) were closed related to NCBI-Blast Babesia bovis (HQ264126.1) with homology sequence identity 97-100% and Babesia bigemina 7.45% were closed related to NCBI-Blast Babesia bigemina (KU206291.1) with homology sequence identity 95-99%, then 43 Babesia species includes (B. bovis and B. bigemina) were submitted into NCBI-Genbank and provided accession numbers (MH503811-MH503853). In conclusion, this study concluded that Phylogenetic tree and homology sequences identity was show accurate in differentiation of Babesia species, and these species can be isolated at from local water buffalo from slaughter house in Al-Najaf city, of Iraq.

Two hundred faeces sample were collected from cattle with different age and sex in Al- Diwaniyah Province. The study was conducted in the period between November 2016 and November 2017. Salmonella typhimurium bacteria identified by routine methods such as culturing on selective media, biochemical test and agglutination test using monovalent and multivalent antisera. PCR was can detection type-1 fimbriae gene coding for fimC of Salmonella typhimurium. Results showed that Salmonella isolates were 14.5% in the bovine fecal samples. Also, the serotyping of isolates by using monovalent and polyvalent antisera revealed that all Salmonella isolates in cows were S. typhimurium. The PCR technique was used for detection of type-1 fimbriae coding gene by specific primer for fimC gene. All S. typhimurium isolates in cows appeared to be contained this gene show one distinct band MW.289 bp when electrophoresed on agarose gel. The results of this score indicated that the PCR technique potentate a loud specify in the disclosing of S. typhimurium especially the serotype that encoded to fimC gene type-1 fimbriae isolated from cows in comparison to other routine diagnostic tests.

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 study was initiated for the first time for identification, using sequencing and phylogenetic analyses, of pseudocowpox PCPV that inhabit dairy cows in Al-Qadisiyah province, Iraq. Scab sampling was performed to obtain specimens from udder and teats of 18 affected cows. Initially, a polymerase chain reaction (PCR) method was followed to target a 408-bp piece of the GM_CSF/IL-2 inhibition factor gene (GIF) that belongs to PCPV. Then, the PCR products were sent out to partial sequencing of the GIF gene. The results of the PCR have indicated the presence of the virus in only 3 out of 18 samples. When the sequences were studied using phylogeny, the results have revealed that one of our PCPV strains has a close matching with some of the world strains such as from New Zealand. While two of the current study strains have clustered together with a strain from Finland. The results of our study confirm the presence of the PCPV in dairy cows that induces milker’s nodules.

Bovine fibropapilloma and papilloma occur in different parts of the skin of animals. Bovine Papillomavirus (BPV) is an oncogenic virus making benign tumor lesion of together mucosal and cutaneous tissue in cattle. In order to confirm the clinical diagnosis; the study planned to make the molecular detection of BPV (DNA) using Polymerase Chain Reaction (PCR) from skin lesions and the phylogenetic analysis. Thirty-eight samples of skin lesions were collected from cattle clinically suspected to be infected with bovine papilloma virus from herds in Al-Qadisiyah Governorate in 2016, the primary clinical diagnosis depended on the morphological appearance and features of the lesion. Deoxyribonucleic Acid (DNA) was extracted from skin lesions; the DNA was examined by PCR technique using specific primer to BPV-1 /L gene-1. Twenty-two samples out of 38 (57,9%), which were collected from different regions in Al-Qadisiyah Governorate, were positive. The sequences of four positive samples of DNA product amplification of (BPV) type-1, L1 gene confirmed the PCR results. These samples had the DNA presented in four accession numbers KY662042-1, KY662043-1, KY662040-1 and KY662041-1. This study proofed that cutaneous bovine papillomatosis related with BPV-1 infection in the cattle herds has affinity to solid skin rather than other epithelial and mucosal tissue.

To detect Anaplasma marginale among carrier cattle by using polymerase chain reaction (PCR) technique, 64 blood samples, fromhealthy cows in abattoir of Al-Nasiriyah city were collected from June till August, 2017 in this study. By targeting MAR1bB2 gene with the molecular weight of approximately 265 bp, Anaplasma marginale were detected in18 samples (28.125%). One of these positive sample was recoded in National Center for Biotechnology Information, NCBI; Gene Bank.

The purpose of this study was to determine the prevalence of clinical, subclinical and chronic infection with the equine parasite T. equi in some Egyptian localities (Cairo and Giza governorates). A panel of 396 equine blood samples representing 141 horses, 250 donkeys and 5 mules was collected from equines during the period from April 2015 to March 2016 using microscopic examination and conventional PCR. Microscopically a twenty two (5.56%) of 396 were positive for T. equi merozoites that appeared as small rounded, pyriform shaped and maltase cross shaped merozoites. Among 8/141(5.67%) horses and 14/250 (5.60%) donkeys were found to have positive for T.equi. A one hundred blood samples (45 horses, 50 donkeys and 5 mules) selected randomly were also examined by PCR. The results of PCR showed 30/100(11/45 (24.4%) horses, 18/50 (36%) donkeys and 1/5 (20%) mule) were positive for T.equi. When the sequenced PCR amplicons (n=3) were aligned to the reference nucleotide sequences of T. equi accessed in Genbank, the horse isolate showed insertion of Thymine (T) base at position 23 and substitution of Thymine (T) base with Cytosine (C) base at position 91, while the donkey and mule isolates have no alterations when compared to the reference sequences. The phylogenetic analysis showed that the sequenced PCR isolates belonged to T. equi. The obtained sequences were deposited in the GeneBank database under accession numbers MF192854, MF192855 and MF192856.

Research on genetic polymorphism of growth hormone (GH) and receptor growth hormone (rGH) has not been done in crossbred of Limousin cattle, so it is interesting to be examined. Blood samples were taken from 14 Madura calves were artificially inseminated with Limousin cement. DNA amplification is done by using Polymerase Chain Reaction (PCR) method, Restriction Fragment Length Polymorphism (RFLP) method to determine the genotype. DNA sequencing was done to determine nucleotide sequences of GH unit genes. The results showed that identification of GH and rGH gene polymorphisms was done by breaking DNA fragments from 432 and 298 bp in Madura and Limousin cattle (Madrasin) ie, L and V alleles have a frequency of 0.67 and 0.33 for the GH gene, respectively. This proves that the crossed-breeding of Madrasin have V allele that is not owned by the Madura cattle. While in the rGH gene, the A allele is 0.92 and the G allele is 0.08, with the frequency of the A allele larger than the G allele. This research concluded: that GH and rGH undergo changes on polymorphisms in Madrasin cattle can be used as a basis for selection.

In the present study, forty donkeys of different ages and sexes at Giza Zoo, Egypt were investigated between October 2015 and September 2016 for the presence of hydatidosis disease. Hydatid cysts were detected in the livers of 10% of the examined donkeys and these cysts had a fertility rate 100%. Female donkeys were infected with cysts more than males and all infected donkeys were old aged with no cases of infection were detected in young or adult donkeys. Using molecular tools, the DNA extracted from cysts that had been isolated was subjected to PCR amplification, using synthesized oligonucleotide primers, and these were constructed to target the 299 bp within the (ND2) gene, which is considered to be specific for the Echinococcus equinus genotype. The sequenced PCR products showed homology to E.equinus (G4 or horse strain genotype). These results can be used in future to pursue the epidemiological status of the causative strain of hydatidosis in equines at the study area.

Theileriosis is parasitic infection causes by obligate intracellular protozoa of the genus Theileria. T. lestoquardi is the most virulent species in sheep and goats which causes a severe disease with a high morbidity and mortality rate. In this study the phylogenetic relationships between two local isolate of T. lestoquardi and nine T. lestoquardi global isolates as well as Babesia ovis out-group isolate were analyzed using the 18S rRNA gene sequence. The multiple sequence alignment analysis and neighbor joining phylogenetic tree analysis were performed by using ClustalW multiple sequence alignment online based analysis of 1098bp 18S rRNA gene was amplified by polymerase chain reaction. Phylogenetic analysis results of these gene sequences revealed that T. lestoquardi local isolates were closely related to T. lestoquardi Iran isolate (JQ917458.1) and two Iraq Kurdistan isolates (KC778786.1 and KC778785.1) more than other countries. This study represents the first report on the use of molecular phylogeny to classify T. lestoquardi obtained in Middle Region of Iraq.

Two groups of broiler chickens were used in this study. One was reared under typical conditions at the animal house of Veterinary Medicine College/Mosul University- Iraq, while the other group was reared under common commercial farm conditions. Fifty and 80 birds from the two respective groups were sacrificed at 49 days of age for detecting Toxoplasma gondii by using Species-specific PCR technique. Results of Latex agglutination test indicated, principally, that 29.3% and 49.2% of the serum samples were positive for the birds of both groups, respectively. Titer figures ranged between 1:20 to 1:320 where the highest value was 1:160 (39.3%) and the lowest was 1:20 (5.8%). Confirmation of 38 and 64 serum samples, using Latex agglutination test was performed by PCR technique, from the two respective groups of chickens. Of those, 8 samples from the college birds and 35 from the commercial farm birds were confirmed positive by giving band of 133 bp, according to specific primers designated on gene B1.Based on these results, pursuing the PCR technique is considered, so far, a most sensitive method for Toxoplasma gondii detection. Also, positive PCR results are counted on as an early marker for reactivation and useful means in monitoring therapies.