Main Subjects : Veterinary Microbiology

Molecular detection of Mycoplasma spp. from camel’s milk

Ammar M. Al-AAlim; Omar H. Sheet; Zahraa M. AL-Jumaa; Mohammad A. Hamad

Iraqi Journal of Veterinary Sciences, 2023, Volume 37, Issue 2, Pages 333-337
DOI: 10.33899/ijvs.2022.134635.2388

The dromedary camels are essential because of their multiple uses for transport, wool, meat, and milk production. Camel’s milk is considered exceptional nutrition, so it uses for therapeutic and high nutrition. Mastitis may affect camel’s milk production, cause significant economic loss, and is associated with zoonotic disease. The study aimed to detect the prevalence of Mycoplasma spp. especially M. bovis in camel’s milk using the polymerase chain reaction (PCR) method as a primary molecular diagnostic technique. Fifty milk samples were collected from she camels suffering from subclinical mastitis in Iraq. The result of the current study declared that 26.66% of the camel’s milk samples were positive for Mycoplasma. In contrast, the prevalence rate of Mycoplasma (M.) bovis in all the samples was 61.53%. The study concludes that Mycoplasma and especially M. bovis are considered one of the bacteria that cause subclinical mastitis in camels, and using the PCR method is regarded as a more rapid, simple, and current for detecting the Mycoplasma bacteria.

Characterization and phylogenetics of beta-lactamase Temoneira gene in Escherichia coli of the Bali cattle on Lombok island, Indonesia

Kholik Kholik; Pudji Srianto; Aulanniam Aulanniam; Fedik Abdul Rantam; Sri Pantja Madyawati

Iraqi Journal of Veterinary Sciences, 2023, Volume 37, Issue 2, Pages 487-493
DOI: 10.33899/ijvs.2022.135062.2441

This study aims to determine the character and phylogenetics of the β-lactamase Temoneira (blaTEM) gene in Escherichia coli isolated from the reproductive tract of Bali cattle with repeat breeder cases. This study was conducted in June 2021 that used 16 female Bali cattle with repeat breeder cases to take their reproductive tract fluid using a plastic sheet gun. Isolation of Escherichia coli, using Eosin Methylene Blue Agar was identified by biochemical tests. Antibiotic susceptibility test on E. coli samples was carried out by disc diffusion method. The screening test for the presence of extended-spectrum β-lactamase was done using the double-disk approximation test method. Characterization of the blaTEM gene in E coli, using the PCR method and sequences of the blaTEM gene, were phylogenetically analyzed. The results showed that 4 E. coli isolates were obtained from 16 samples of reproductive tract fluid. The sensitivity test to antibiotics of 4 isolated samples of E. coli showed that 50% were resistant to penicillin and cefotaxime. Samples of E. coli resistant to penicillin and cefotaxime showed synergy results in the double-disk approximation test. The PCR results showed that the samples of E coli encoded the blaTEM gene which was located at the 560 bp position on gel electrophoresis. The phylogenetic tree analysis found that the blaTEM gene encoded by E. coli was related to E. coli encoding the blaTEM-206 gene. The character blaTEM gene in samples of E. coli showed the character of blaTEM-206 gene of extended-spectrum β-lactamase-producing E. coli. Escherichia coli from the reproductive tract of Bali cattle showed the character of E. coli S2.2-EK pEC-S2.2 blaTEM gene for class A broad-spectrum B-lactamase TEM-206 with a high degree of kinship.

Mycoplasma from the upper respiratory tract and conjunctival infections in household dogs

Saba A. Hussein; Mohammad A. Hamad

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue Supplement I, Pages 137-141
DOI: 10.33899/ijvs.2022.135824.2525

The study was performed to isolate and detect Mycoplasma as a causative agent of the upper respiratory tract and conjunctival infections in household dogs in Mosul city. One hundred household dogs of different ages, sex, and breed were subjected to the study, including 60 dogs suffering from moderate to severe respiratory infections and 40 healthy dogs in Mosul city from 1/2/2022 to 15/6/2022. Three hundred swabs were collected, including nasal swabs 100, oropharyngeal swabs 100, and conjunctival swabs 100. The swabs were cultured in Mycoplasma media and incubated at 37ºC with 5% CO2 in a candle jar for 4-14 days. Light and the dissecting microscope examined the growing colonies microscopically under low magnification. The colonies were also stained with modified Dienes stain. The results of the current study indicated that respiratory infections were predominant in young male household dogs compared with conjunctival infections in all examined dogs. The conjunctival infections were obvious only in Husky and Belgium dogs. The high isolation rate of Mycoplasma was from upper respiratory tract infections in diseased female dogs more than one-year age 100%. In contrast, the conjunctival infections were more dominant and had a higher isolation rate in less than one-year males. The current study revealed a high Mycoplasma isolation rate in respiratory swabs of Terrier and German dogs, while the conjunctival swabs were positive for mycoplasmal culture in Husky and Belgium dogs 100%. In conclusion, mycoplasmal infections were more dominant in upper respiratory infections in female German and Terrier dogs.

Immune response strategies of Brucella melitensis and their antigens in rats

Basher S. Noomi; Sanaa S. Ahmed; Hiba Y. Khalaf; Nihad A. Jafar

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue Supplement I, Pages 27-30
DOI: 10.33899/ijvs.2022.134990.2431

Brucella melitensis is an intracellular bacterium and is the main brucella species that cause abortion and placenta retention in sheep and goats. It has many mechanisms to evade the immune response. The current study aimed to investigate Brucella melitensis strategies for producing immune responses in rats after challenging the bacterium. For this purpose, live and killed Brucella melitensis REV1 strain was given to rats subcutaneously, and immunological markers like TLR2, TLR4, IFN- γ, and anti-brucella antibodies were determined. The results showed that the level of immunological markers like TLR2 and TLR4 did not significantly increase in rat groups inoculated with live Brucella melitensis, while it increased in the rats’ groups vaccinated with the sonicated Brucella melitensis; also, the results showed an increase in the level of IFN-γ and anti-brucella antibody titers in all animal groups. The study concluded that the inoculation with killed bacteria and REV1 could protect the animals against challenging doses, as seen when the groups were inoculated with challenge dose of the bacterium.

Morphological detection of dermatophytes isolated from cattle in Wasit province

Dhurgham A. Alhasan; Hawraa F. Al-Abedi; Talal J. Hussien; Atheer Q. Mohammad Ali

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue Supplement I, Pages 167-172
DOI: 10.33899/ijvs.2022.135833.2530

This study aimed to diagnose the skin lesions in cattle that were suspected to be ringworm in order to determine the species causing these lesions. A total of 40 samples of skin desquamation and hair lesions were collected from cattle in a farm located in the Wasit province during the period from October 2021 to March 2022. Diagnosis used to be carried out the usage of direct microscopy of potassium hydroxide, isolation of the fungus through the use of Sabouraud dextrose agar supplemented with chloramphenicol and cycloheximide, as well as by way of detecting those fungi through staining with Lactophenol cotton blue. The diagnosis results revealed that infection of cattle with ringworm represented 20/40 (50%) of skin lesions and the majority of the common dermatophytes spp was identified as Trichophyton mentagrophytes 15/20 (75%) and Microsporum canis5/20 (25%) depending on the fungal growth. In conclusion, the most common skin lesions of cattle in Wasit province were identified as ringworm.

Molecular identification of virulence genes of Pseudomonas aeruginosa isolated from fish (Cyprinus carpio) in Mosul city

Asmaa J. Altaee; Sumaya Y. Aldabbagh

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 4, Pages 953-958
DOI: 10.33899/ijvs.2022.132660.2119

Depending on the nature of the fish's environment, they are susceptible to many pathogens, including bacterial causes, so the goals of the current study are isolation and molecular identification of Pseudomonas aeruginosa with its prevalence rate and detected virulence genes from fresh common carp fish. The swabs were taken from the gills, skin, intestine and muscles of 75 fish samples from variable localities from Mosul city during the period September to December in the year 2021. The prevalence percentage of bacteria was 26.66% which was confirmed by traditional microbiological tests which included (phenotype culture, microscopically features and API-test) and molecular identification. The isolates formed 42.5, 37.5, 15, 5% from gills, skin, intestine and muscles, respectively. The molecular results of forty isolates determine that Pseudomonas aeruginosa have rpoB 100%, and virulence genes oprL, toxA, and algD, which are express the outer membrane protein, exotoxin A and alginate respectively occur as 97.5% for the oprL gene and 100% of both toxA and algD genes.

Multi-drug resistant phenotypes of extended-spectrum β-lactamase (ESBL)-producing E. coli from layer chickens

Adewale Olopade; Asinamai A. Bitrus; Asabe H. Halimat Momoh-Zekeri; Pwaveno H. Bamaiyi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 4, Pages 945-951
DOI: 10.33899/ijvs.2022.132655.2117

Antimicrobial resistance (AMR) is a growing and emerging public health problem worldwide. This research determines the occurrence of ESBL E. coli and antimicrobial resistance profiles of E. coli on eggshells from selected layer chickens. The shells of 270 egg samples were swabbed to detect the presence of E. coli. E. coli isolates were recovered from 73(23%) of the 270 samples collected. The isolates were subjected to antimicrobial susceptibility testing using six panel antibiotics (ampicillin, tetracycline, sulphamethoxazole-trimethoprim, gentamicin, imipenem, and ciprofloxacin) using the disk diffusion method. The isolates showed the highest resistance to Ampicillin 95.9%, closely followed by tetracycline 89%, sulphamethoxazole-trimethoprim 72%, gentamicin 41.1%, and imipenem 1.4%. Also, 78% of the isolates were multi-drug resistant. A 56/73 (76.7%) out of seventy-three isolates were screened as presumptive ESBL-E. coli by culture on ESBL CHROM agar and 42/56 (75%) of the isolates yielded ESBL-producing E. coli based on the production of ESBL by double disc diffusion method. The questionnaire survey results showed that all farms used antimicrobial agents for therapeutic or prophylactic purposes. Also, not all the farms had suitable biosecurity measures. The findings of this study indicated that eggshells are potential reservoirs for multi-drug resistant E. coliand ESBL-Producing E. coli.

Phylogenetic tree constructed of Salmonella enterica subspecies enterica isolated from animals and humans in Basrah and Baghdad governorates, Iraq

Maitham S. Sadiq; Rasha M. Othman

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 4, Pages 895-903
DOI: 10.33899/ijvs.2022.132478.2096

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.

Effect of garlic oil nanoemulsion against multidrug resistant Pseudomonas aeruginosa isolated from broiler

Amal S. El-Oksh; Dalia M. Elmasry; Ghada A. Ibrahim

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 4, Pages 877-888
DOI: 10.33899/ijvs.2022.132430.2094

This work aimed to focus on the antibacterial properties of garlic nanoemulsion on some multidrug resistance (MDR) strains of Pseudomonas aeruginosaisolated from broiler farms and hatcheries in Sharkia and Ismailia governorates, Egypt. Pseudomonas spp. was isolated in 21.3% of collected samples. It was isolated from younger broilers 1-10 days with an incidence rate of 22% (11/50), older broilers 16% (8/50), dead embryo in shell 31.4% (11/35), and from hatcheries was 13.3% (2/15). There was a variable range of antibiotic resistance ranging from 66.7-100% against the isolated strains of P. aeruginosa. Tetracycline and sulfamethoxazole-trimethoprim achieved the highest resistance rates, while penicillin and gentamycin were of a lower rate. However, norfloxacin, ciprofloxacin, and colistin were the most sensitive antibiotics against examined MDR P. aeruginosa.16SrDNA gene was found in ten P. aeruginosa isolates. Theseisolates were found to be virulent as oprL gene was detected in all isolates 100%. In addition, tetA(A),blaTEM,arr, and mexRantibiotic resistance genes were shown positive 100% in all MDR P. aeruginosa isolates. Minimum Inhibitory Concentration (MIC) values showed that garlic nanoemulsion (GN) was effective against examined P. aeruginosa at different concentrations. GN had 29.61% sulfur compounds of active components with 0.52 ug/ml of IC50 and 40.94 nm size with polydispersity index: 0.165 using dynamic light scattering had a 19.6± 5.11mV. In conclusion, the application of garlic nanoemulsion is an excellent alternative candidate to antibiotics for treatment because it significantly reduced the gene expression levels of MDR P. aeruginosa in broiler farms.

Control of biofilm-producing Aeromonas bacteria in the water tanks and drinkers of broiler poultry farms using chitosan nanoparticle-based coating thyme oil

Asmaa N. Mohammed; Amira S. Attia

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 3, Pages 659-669
DOI: 10.33899/ijvs.2021.131253.1935

In a drinking water distribution system, biofilm-producing bacteria are considered an alarm bell for increased emergence of waterborne pathogens. This study aims to monitor the prevalence of biofilm-forming Aeromonas species in the drinking water distribution systems in different broiler chicken farms. The antimicrobial activity of thyme essential oil (TEO), thyme essential oil nano-emulsion (TEO-N), chitosan (CS), chitosan nanoparticles (CS-NPs), and both CS and CS-NP-based coating TEO against the different Aeromonas spp.was evaluated using the broth microdilution and agar well diffusion assay. The overall prevalence rate of Aeromonas spp. was 49.3% (74.0/150). The highest rate of Aeromonas isolates was noted in water drinkers and tanks 75.0% (30/40) and 62.5% (25/40), respectively) followed by feedstuff 40.0% (12/30). In contrast, the highest percentage of biofilm-producing Aeromonas spp. was Aeromonas hydrophila 70.0% (14/20) followed by Aeromonas caviae 30.0% (6/20). The fatal effect of CS-NPs against all isolated Aeromonas spp. was achieved 100% at 1.5 and 2.0 µg/mL. Moreover, chitosan nanoparticles coating thyme essential oil (CS-NPs/TEO) verified the lethal effect 100% on both A. hydrophila and A. caviae at the ratio of 1:1 and 1:0.75 µg/mL. In conclusion, the main source of Aeromonas spp. in the drinking water distribution system was the unhygienic status of water tanks and drinkers that allowed biofilm to produce due to aggregation of Aeromonas bacteria on the inner surface of that equipment. Both CS-NPs and CS-NPs/TEO could be applied as a sanitizer and/or disinfectant for Aeromonas biofilm control.

Detection of Campylobacter fetus in aborted ewes in Sulaimani province by PCR

Eman D. Arif

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 3, Pages 647-651
DOI: 10.33899/ijvs.2021.131225.1931

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.

Secondary bacterial infection of hydatid cysts infected livestock animals (In vitro study)

Maimonah Q. Yahya; Zahraa Khairi-Aldeen Mohi-Aldeen

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 3, Pages 761-768
DOI: 10.33899/ijvs.2022.131873.2016

Isolation and identificationof the bacteria-infected hydatid cystsof livestock animals were carried out. The study lasted for five months. A total of 302 slaughtered livestock were checked to find hydatid cyst naturally infected livers and lungs of sheep and goat at local Mosul abattoir/ Nineveh/ Iraq. Each cysts' aspirated fluid was cultured separately on blood or nutrient agar at 37C for 24 hours then detected secondary bacteria. Each bacterium was incubated independently with 1000 viable protoscolices in a culture tube containing tryptic soy broth at 37°C and examined every two hours for 6 hours, followed by 12, 24, 36, and 48 hours afterward.The degeneration rate of protoscolices resulting from treatment with bacteria isolated depending on viability was studied in vitro.The results were revealed that the liver was the preponderant affected organ in sheep and goats. About 57.1% of sheep and 50% of goats harbored fertile cysts. Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Proteus mirabilis, and Pseudomonas aeruginosa bacteria were observed in the infected cysts. The most common bacteria infect the cyst was Escherichia coli. The protoscoleces treated with the bacterial isolates had completely deteriorated, whereas 97% of protoscoleces throughout the control groups were still alive and intact even after completing the incubation time. The experiments detect a significant time-dependent scolicidal effect on decreased viability of protoscolices in vitro study. This could pave the door for more research into the scolicidal validity of bacteria or their by-products as protoscolices both in vitro and in vivo.

Molecular detection of ESBL/AmpC ß-Lactamase Escherichia coli isolated from sheep in Mosul city

Fatma R. Rafee Mahmood; Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 387-392
DOI: 10.33899/ijvs.2021.130380.1810

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.

Molecular characterization of extended spectrum beta-lactamase producing Klebsiella pneumoniae isolated from cows in Mosul city, Iraq

Sumaya Y. Aldabbagh

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 375-380
DOI: 10.33899/ijvs.2021.130341.1803

Cows are considered as reservoir hosts for many pathogenic bacteria that are resistant to broad-spectrum beta lactamase(ESBL). Presence of ESBL resistant K. pneumoniae in nasal of cows and beef meat constitutes a risk for public health due to transfer of antibiotic resistance gene from cows to environment, humans and farm animals. Therefore, the current study was concluded the detection of molecular characteristics of ESBL producing Klebsiella pneumoniae that was isolated from both cow's nostrils and from local beef samples. Fifty nasal swabs were collected from farms cows in Mosul city, and 50 samples of beef from local butcher shops for the period from February to August 2020. Bacterial isolation and identification tests were conducted for ESBL resistant Klebsiella pneumoniae using MacConkey agar medium with-1 μg/ml cefotaxime. PCR was carried out to confirm the results using special primers (SSKP 1 F and SSKP 1 R) for Klebsiella (the target gene 16srRNA). Then, a molecular examination was performed using the precursors CTX-M, TEM and SHV. Through bacterial isolation, 36 (72%) and 28 (56%) isolates were belonging to ESBL resistant Klebsiella pneumoniae from nasal samples and beef meat respectively. The CTX-M, TEM and SHV genes formed (100, 89.2, 85.7) % and (100, 72.2, 71.4) % for each the meat and nasal samples respectively. This study showed that cows play a major role in transferring ESBL producing Klebsiella pneumoniae from cows to humans as a result of environmental handling or consumption contaminated meat. 

Some immunological responses in rats injected with prepared bacterin toxoid of local methicillin resistant Staphylococcus aureus

Abdullah A. Abdulazeez Sheehan; Mohammed H. Khudor; Fanar A. Isihak

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 401-406
DOI: 10.33899/ijvs.2021.130401.1815

This study was aimed to prepare bacterin toxoid from local isolate of methicillin resistant Staphylococcus aureus (MRSA) isolated from bovine mastitis in Basrah province. Isolated MRSA was detected and confirmed using polymerase chain reaction. Then inactivated bacterin toxoid was prepared. A total of 45 male albino rats were used in 3 mainly groups to assess the bacterin toxoid. For immunological response, the concentrations of IL-4, CD4+, and CD8+ T-cells were estimated using enzyme linked immunosorbent assay. The results showed increase in the concentrations of IL-4, CD4+ and CD8+ T-cells in vaccinated group comparing with non-vaccinated control group. Increasing concentrations of IL-4 enhance humoral immune response by stimulation of Th2 that effect directly on B-cells differentiating them to plasma cells responsible for production of specific antibodies against MRSA. Increased CD4+T-cells also enhance humoral immune response as a result of interaction between them and antigen-presenting cells which presented major histocompatibility complex (MHC) type II on their surface, while increased CD8+T-cells enhance cellular immune response as a result of interaction between them and somatic effected cells presented MHC type I on their surface leading to differentiation them to Cytotoxic T lymphocytes (CTLs) responsible for the killing of effected cells. It was concluded that locally prepared bacterin toxoid proved their efficacy to stimulate both humoral and cell mediated immunity in rats as an experimental animal model.

Phenotypic characterization and antibiogram of extended spectrum ß-lactamase (ESBL)/AmpC-producing Escherichia coli isolated from sheep

Fatma R. Mahmood; Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 2, Pages 303-307
DOI: 10.33899/ijvs.2021.130112.1732

This study aimed to isolate and characterize extended-spectrum ß-lactamases (ESBLs) and AmpC producing E. coli in sheep in Mosul city. A total of 260 milk and fecal samples were collected aseptically from healthy ewes (n=60), their respective lambs (n=60), and ewes with clinical mastitis (n=40). Standard bacterial isolation and identification on special culture media were performed to isolate ESBL/AmpC producing E. coli. While special antibiotic discs D68C MASTDISCS® Combi AmpC and ESBL ID set were used to characterize positive ESBL/AmpC E. coli. The results showed that 99/260 (38.1%) of tested samples were ESBL-E. coli positive and distributed as follows, 7/60 (11.7%) and 39/60 (65%) from milk and feces of clinically healthy ewes, respectively, and 37/60 (61.7%) from feces of clinically healthy lambs, while 4/40 (10%) and 12/40 (30%) from the milk and feces of ewes with clinical mastitis, respectively. However, we could not obtain any AmpC positive isolate from all tested samples. The high recovery percentages of ESBL from feces or milk of sheep reflect the arbitrary use of the antibiotic in sheep farming. This could significantly increase the resistance of the bacterial population that might represent a potential source for transmission of antibiotic resistance to humans.

Molecular characterization and phylogenetic analysis of Escherichia coli isolated from milk of cattle affected by mastitis

Azhar A. Neamah; Khilood H. Fahad; Jenan N. Sadeq; Monyer A. Al-Fatlawi

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 251-254
DOI: 10.33899/ijvs.2021.129934.1702

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.

Extraction and purification of lipopolysaccharide from Escherichia coli (local Isolate) and study its pyrogenic activity

Ammar M. Al-AAlim; Ali A. Al-Iedani; Mohammad A. Hamad

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 45-51
DOI: 10.33899/ijvs.2021.128963.1614

In this study, we tried to extract and purify the LPS from E. coli local isolate and determine the molecular weight, purity, and pyrogenic effect of the product and compare it with standard E. coli O55:B5 LPS, the E. coli LPS was extracted by using hot phenol method then SDS- PAGE was used with both Coomassie blue and silver nitrate stain to determine its molecular weight and protein contamination also we used HPLC to the estimation of E. coli LPS purity and finally the pyrogenicity of extracted E. coli LPS was tested by using rabbit pyrogen test. The result showed that the hot phenol method with enzymatic treatment gave highly pure LPS with a high yield reach up to 242.4 mg, staining the SDS page gel with Coomassie blue and silver nitrate uncover the high purification of the extracted LPS (ELPS) with no protein contamination, with a molecular weight range between 15-23 kDa, HPLC test reveals that purity of ELPS was 100 % compared with standard LPS. The rabbits' pyrogen test confirmed that the biological activity of ELPS. In conclusion, the LPS was extracted with high purity compare with standard LPS and without any protein or DNA contamination by using the hot phenol method also the extracted rough LPS was slightly lighter than the standard LPS used but this did not affect its biological activity which remained intact 

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

Saba F. Klaif; Asaad J. Abid; Monyer A. Al-Fatlawi; Mansoor J. Ali

Iraqi Journal of Veterinary Sciences, 2022, Volume 36, Issue 1, Pages 85-88
DOI: 10.33899/ijvs.2021.129230.1635

This study was purposed for confirming detection and typing of Anaplasma spp in infected sheep from Al-Diwaniyah province, Iraq. Sampling of 50 blood specimens was performed from clinically-identified infection of anaplasmosis. The samples of the blood were subjected to DNA extraction followed by polymerase-chain-reaction (PCR)-based detection of the Anaplasma marginale using major surface protein (MSP4) gene. The results have shown that 8 blood samples were infected with A. marginale. The PCR-based identification has revealed a confirmative identification of the Anaplasma marginale in the infected sheep. The study identifies Anaplasma marginale as a member of infectious agents that affect sheep in the study city. 

Immunotherapeutic effect of chitosan and listeriolysin O on Listeria monocytogenes infection in mice

Hero I. Ali; Shaimaa N. Yasssein

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 149-155
DOI: 10.33899/ijvs.2021.132332.2082

This study aimed to evaluate the effect of dietary chitosan and listeriolysin O (LLO) on the immune response against Listeria monocytogenes infection in mice. L. monocytogenes isolate was obtained from the unit of Zoonotic Disease, College of Veterinary Medicine, University of Baghdad. It was reactivated and cultured on PALCAM agar and exposed to a confirmatory diagnostic test. listeriolysin O (LLO) was extracted and purified. Sixty mice were used and divided into four groups each one involving 15 mice; the first group was fed on diet supplemented with a chitosan for 4 weeks. At the end of the 2nd week, normal saline was injected S/C; the second group was fed on a normal diet without any addition. At the end of 2nd week, 0.3 ml of LLO was injected S/C, then the booster dose of LLO was given after 14 days of 1st injection; the third group was fed on a diet supplemented with chitosan as in G1, and treated as in G2 and the fourth group fed on a normal diet without any addition and treated as in G1. A skin test was performed on 5 mice of each group while a Challenge test by injection of 0.2ml of 1x109 CFU/ml of viable L. monocytogenes intraperitoneally was performed on other mice. The concentration of IgG titer and IL6 were measured. The results revealed that the third group recorded significantly higher values in the skin thickness, IgG, and IL6 concentrations when compared with other groups at different times which indicate that LLO and chitosan may support each other to provide the most beneficial effect by eliciting of good cellular and humeral immunity against listeriosis.

Multi-drug resistance profile of extended spectrum β-Lactamases producing Escherichia coli isolated from sheep in Mosul city

Fatma R. Mahmood; Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 37-40
DOI: 10.33899/ijvs.2021.130475.1829

Multi-drug resistance (MDR) Escherichia coli have become a major threat due to their ability to overcome different types of antibiotics. However, Extended Spectrum β-lactamase E. coli (ESBLE) imposes an additional threat due to their ability to resist the 3rd generation cephalosporins. Accordingly, our study aimed to investigate the antibiogram profile of ESBLE isolates obtained from sheep. A Total of 40 ESBLE isolates were included in this study which represents sheep fecal and milk samples. Twelve antibiotics were selected to perform antibiotic sensitivity tests following standard microbiological methods. The results of the study showed that the highest resistance percentages were recorded for tetracycline 97.5%, ciprofloxacin 80%, trimethoprim/sulfamethoxazole 65%, and streptomycin 57.5%. While other antibiotics recorded lesser values. On the other hand, all isolates were susceptible to gentamycin and tobramycin each at 92.5%, followed by chloramphenicol and levofloxacin each at 82.5% and nitrofurantoin 72.5%. While fewer values of sensitivity were recorded for streptomycin, trimethoprim/sulfamethoxazole, azithromycin, nalidixic acid, ciprofloxacin and, tetracycline. The study concluded that ESBLE of sheep origin that have additional resistance features to other antibiotics could be a major threat for spreading resistance and contaminating the environment and finally impose negative impact for response to antibiotic treatment in humans.

Isolation and antimicrobial resistance of Staphylococcus spp., enteric bacteria and Pseudomonas spp. associated with respiratory tract infections of sheep

Teba A. Aziz; Inam J. Lafta

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 53-58
DOI: 10.33899/ijvs.2021.131098.1917

Sheep are considered as an important part of livestock in the worldwide, particularly in Iraq, as they provide meat, milk, leather, wool, and manure. The present study aim is isolation and identification of staphylococci, enteric bacteria and Pseudomonas spp. Totally, 115 samples were collected from sheep (100 samples were collected from the nasal cavity of local sheep suffering from respiratory infections, and 15 samples were collected from apparently healthy local sheep). All the samples were collected from seven flocks located in Abu Ghraib and Al-Radwaniyah, Baghdad governorate, Iraq. The samples were taken during the period from October 2020 to February 2021. Staphylococcus spp., Pseudomonas spp., andenteric bacteria were detected firstly by using classical diagnostic methods, and secondly they were identified to the species level by using the corresponding analytical profile index (API 20 staph, API 20NE and API 20E) and via the Vitek2 system. Importantly, from nasal swabs, 79% bacterial isolates were obtained, including Enterobacter cloacae, Escherichia coli, Raoutltella planticola, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus among others. The antimicrobial susceptibility test showed multi-drug resistant S. aureus, P. aeruginosa and K. pneumoniae. To conclude, several aerobic bacteria were isolated from the nasal cavity of diseased sheep. An extensive study is needed to determine the economic and public health impacts of these bacteria.

Isolation and molecular detection of enterotoxigenic Staphylococcus aureus from raw milk of cows

Dunya D. Taher; Safana A. Yassin; Mariam H. Abdulkareem

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue Supplement I-III, Pages 137-141
DOI: 10.33899/ijvs.2021.131957.2030

Staphylococcus aureus is a microbe associated with human’s food poisoning. It is caused by isolates producing different heat stable enterotoxins which act as one of the most spread worldwide gastroenteritis causes. The goal of the current study was to assess the incidence of S. aureus (enterotoxigenic) in the raw milk samples of cows which were collected from various places of Baghdad city. Isolation and identification of S. aureus were done by conventional laboratory method and the diagnoses were confirmed by using molecular method RT-PCR to detect the specific gene of S. aureus nuc gene. In addition, we investigate the occurrence of some of Staphylococcal enterotoxins genes such as SEA, SEB, SEC, SED, and SEE. We found that 12/50 (24%) of the isolates were S. aureus and these isolates carried one or more of the enterotoxin genes such as enterotoxin type SEC 12/12 (100) followed by enterotoxin type SEA 7/12 (58.33%) and only 1/12 (8.33%) was type SED, while type SEB and SEE were negative 0%. Some of the isolates have genes that coded for two enterotoxins patterns such as SEA and SEC 7/12 (58.33%) and SEC and SED 1/12 (8.33%). Fewer of the isolates expressed only one genotype of enterotoxin gene like SEC 4/12 (33.3%). This study has proven that there was a high chance of occurrence for the enterotoxigenic S. aureus at the raw milk of cows in Baghdad city. 

Genetic diversity and clonal relatedness of Aeromonas hydrophila strains isolated from hemorrhagic septicemia’s cases in common Carp (Cyprinus carpio) farms

Zanan M. Taha

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 4, Pages 643-648
DOI: 10.33899/ijvs.2020.127566.1511

The objective of this study was to determine molecular typing and comparison analysis of 24 Aeromonas hydrophila isolated from the diseased fish with hemorrhagic septicemia in freshwater ponds and cage in Mosul and Duhok cities, Iraq. A total of 24 A. hydrophila isolates that were collected from various fish ponds and cage, were used in this study. Identification of isolates was made by the standard microbiological and molecular methods. ERIC-PCR was done with different primers to establish the genetic relationship between strains. ERIC-PCR typing showed that 24 strains of A. hydrophila were classified into 11 ERIC types (genotypes). Genotypes 9 and 7 represented the most prevalent clone. All A. hydrophila strains that were isolated from the same fish were genetically diverse. There was minimal genetic similarity between some strains which were retrieved from the same geographical source area. Also, some isolates from different geographic source area were showed a 100% genetically similar. Aeromonas hydrophila was genotypically heterogeneous and clonally dispersed among different fish ponds and cage in Mosul and Duhok cities, Iraq. Besides, one fish can be infected with more than one strains of A. hydrophila.

Molecular characterization of extended spectrum cephalosporin resistant Escherichia coli isolated from dogs

Ihsan M. Ahmed; Sumaya Y. Aldabbagh; Dhyaa M. Jwher

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 3, Pages 473-478
DOI: 10.33899/ijvs.2020.127032.1441

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.

Molecular analysis of ompA gene Pasteurella multocida Indonesia local isolates

A. T. Soelih Estoepangestie; Arini Rahmi Dewi; Suwarno Suwarno; Didik Handijatno; Rahaju Ernawati; Wiwiek Tyasningsih

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 211-216
DOI: 10.33899/ijvs.2019.125934.1191

The aim of this research was to analyze ompA molecular gene of Pasteurella multocida buffalo isolate and bovine isolate from Nusa Tenggara Timur, Indonesia and Katha strain isolate from hemorrhagic septicemia vaccine. Determinant of P. multocida local isolates ompA gene amplification sequencing PCR then conducted to see the sequence of nucleotide sequences of ompA gene. The results of PCR amplification showed an amplicon of 559 bp of all isolates. The homology analysis result of the isolates ranged from 93 - 100% with 13 P. multocida isolates from GenBank, and phylogenetic tree analysis shows that buffalo isolate was closely related to Katha strain, Iran, India and China isolate. Whereas bovine isolate far enough with buffalo and Katha strain isolate. Nucleotide sequences were compared to amino acids then by the method of Kolaskar and Tongaonkar antigenicity predicted antigens in P. multocida. B cell epitope predictions from local isolates and Katha strain were found in five peptides QVSPVFAG, IPELALRVEYQ, GQSVYVPEVVSKT, LKSASVAVAG, and ANYLVAKG.

Detection of CTX-M gene in extended spectrum β-lactamases producing Enterobacteriaceae isolated from bovine milk

Ihsan M. Ahmed

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 397-402
DOI: 10.33899/ijvs.2020.126909.1412

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.

Study the effect of cloned pET-32a(+) plasmid by Lysostaphin gene against Staphylococcus aureus

Jalal Yseen Mustafa

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 233-238
DOI: 10.33899/ijvs.2020.126698.1362

Lysostaphin is a protein zinc metalloproteinase, extracted from Staphylococcus simulans, which disrupting peptide layer of S. aureus. In this study, Lysostaphin gene was detected in the S. simulans isolates. The molecular weight of the Lysostaphin gene is 750 bp. We were used the pET-32a(+) plasmid to cloning lysostaphin gene which transformed to competent rubidium chloride E. coli DH5α for producing the lysostaphin protein. The lysostaphin protein from this gene which isolated from S. simulans, then used the expression of used to killed S. aureus, which has the thick layer of wall that is the very difficult bacteria response to treatment. The result was reported succeeded pET-32a (+) plasmid to expressed lysostaphin gene and gave lysostaphin protein with high quality and quantity. As well as the result was appeared the high accuracy of his tag method in protein extraction and purification, and the quality and quantity more than other studies.

Investigation of gcat gene and antibiotic resistance pattern of Aeromonas hydrophila isolated from hemorrhagic septicemia’s cases in fish farms

Zanan M Taha; Shaaban Tayar Sadiq; Warheen Aziz Khalil; Kaheen Yusif Muhammad-Ali; Hayam Salih Yosif; Hozan Nizar Shamil

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 2, Pages 375-380
DOI: 10.33899/ijvs.2020.126876.1405

The significance of Aeromonas hydrophila concerning hemorrhagic septicemia in aquaculture farms production in Duhok province, Iraq was investigated. Antibiotic-resistant profiles of isolates were also investigated with 8 antibiotics. Bacterial isolates were identified by using morphological and biochemical tests and confirmed molecularly by amplification of gcat gene. Out of 25 examined fish, only 19 fish were harbored A. hydrophila. Twenty-four A. hydrophila strains were isolates from 100 organ samples. Ninety-six percentages of the isolates were resistant to each of the imipenem and gentamicin, followed by doxycycline 92%, ciprofloxacin and trimethoprim-sulfamethoxazole 88%, norfloxacin 58% and ceftriaxone 33%. None were resistant to levofloxacin. Eighty-eight percentages were multiple antibiotics resistant. The high isolation rate of A. hydrophila in our study indicates that this species was the major cause of the outbreak in hemorrhagic septicemia’s cases in our area affecting carp farms and the high rate of resistance should be considered as these isolates can serve as a resistance source for human being during food series and make a great challenge for their therapeutic opportunity.

Incidence of internal parasites of the slaughtered local breeds of ducks and geese

Baydaa Younis Al-lahaibi; Manal Himmadi Hasan; Ahlam Fathi Altaee

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 39-44
DOI: 10.33899/ijvs.2020.126242.1272

This study was carried out to determine the presence of gastrointestinal parasites of local ducks and geese in Nineveh province. Sixty-four ducks and seventy geese of different ages and sexes were purchased from local markets. Necropsy findings in ducks reviled a total infection rate of 68.8% was with protozoa, 50% was with nematodes, while 28.1% were with cestodes. On the other hand, in geese, the percentages with the mentioned parasites were 78.6% with protozoa, 54.2% with nematodes, 31.4% with cestodes. Four types of nematodes were identified in ducks; Ascaridia galli, Heterakis gallinarum, Heterakis isolonche, and Subulura brompti, the same were also found in geese except Heterakis.isolonche. Cestodes identified in ducks and geese were Railletina tetragona, Railletina echinobothrda, Railletina cesticillus and Coantaenia infundibulum. The detected protozoa include Eimeria spp., Tyzeria spp., Wenyonella spp., Cryptosporidia spp., Giardia spp. Double infection with parasite was higher in ducks while the triple infection in geese was the higher.

Physiological effects of lactic acid bacteria against melamine induced toxicity in female albino rats

Ali H. Jameel; Mohammed J. Mohammed; Manal S. Mahdi; Karkaz M. Thalj

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 1-7
DOI: 10.33899/ijvs.2020.126183.1259

The aim of this study was to investigate the ameliorative effect of two type of lactic acid bacteria Lactobacillus casei and Lactobacillus acidophilus against melamine toxicity by some physiological indicators in mature female rats after 21 days. In this study using 35 of female mature rats and divided randomly into seven groups each group contain five animals. The results showed that melamine caused a significant decrease in the organs weights liver and spleen and increase in kidney weight with increase of melamine concentration. Also showed to decrease in value of hemoglobin, red blood cells, white blood cells, lymphocyte and platelets, while the values of granules were increasing with increase of melamine concentration as compared with control group. Also found that the addition of melamine led to increase in cholesterol, low density lipoproteins and blood glucose, while the values of triglyceride and high density lipoproteins was decreased with increase of melamine concentration. The addition of two types of lactic acid bacteria L. casei and L. acidophilus led to decreasing the negative effect of melamine on the values of all the parameters determined.

Molecular fingerprinting of methicillin resistant Staphylococcus aureus strains isolated from human and poultry in Duhok, Iraq

Hishiyar A Hado; Mahde S Assafi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 99-103
DOI: 10.33899/ijvs.2020.126375.1310

Methicillin-resistant Staphylococcus aureus (MRSA) has been recently identified in poultry and farm workers. The aim of this work was to investigate the epidemiological relatedness of MRSA among chickens and farmworker. MRSA isolates (n=50) from human (n=14) and from chikens (n=36) were tested for molecular epidemiological relatedness between human and poultry. RAPD-PCR was carried out for fingerprinting of MRSA isolates genome. Seven genotypes group (A-G) have been identified. All human MRSA were belonging to genotype A. Whereas, chickens MRSA isolates was belonging to different genotype patterns groups (A-G). To conclude, human MRSA was belonging to one genotype pattern but the chickens MRSA strains were belonging to seven genotypes. The genotype pattern A was the most dominant among all MRSA isolates. It is possible that the chickens play an important role for the human exposure to MRSA by direct contact. Further studies are required to address the relatedness between human and chicken MRSA.

Evaluation the safety and synergistic effect of NiFe2O4 nanoparticles with antibiotic against Pseudomonas aeruginosa

Hala M. Majeed; Heba Younis Khalef; Halah Abdulkhaleq Awadh; Bashar Sadeq Noomi; Nihad Abdul-Hussain jafar; Khaild Ahmad Hadi

Iraqi Journal of Veterinary Sciences, 2021, Volume 35, Issue 1, Pages 71-77
DOI: 10.33899/ijvs.2020.126298.1294

The antimicrobial resistance currently impedes and threatens the future of effective prevention and treatment of the continually expanding range of infections caused by bacteria. This study aimed to identify the bacterial causes the wound infection among animals and using the antibiotic/nanoparticles mixture as a new attempt for the treatment the wound infection induced in rats. For this purpose, 112 swabs wound infection cases in the different animal types (36 sheep, 21 goats, 12 cows, 4 horses, 8 dogs, 9 rabbits, 7 genies pigs and 15 rats) were studied in the for bacterial isolation. The Pseudomonas aeruginosa was tested for its sensitivity to the antibiotics and the nanoparticles (CoFe2O4 and NiFe2O4) in vitro by using the MIC method. Also the wound infection was induced in the rats and the effect of nanoparticles/antibiotics mixture were tested in vivo. The results showed that P. aeruginosa was the predominant bacterial type that the caused wound infection. The minimum inhibitor concentration of NiFe2O4 and CoFe2O4 nanoparticles were 32 µg /ml and 16 µg /ml respectively. A clear synergistic effect of antibiotic/ nanoparticles as antibacterial were noticed which appear as a decrease in MIC and increase of the inhibitory diameter zone. According to the result of Random Amplification of Polymorphic DNA test, the nanoparticles effects on genetic material of P. aeruginosa observed as an appearance/disappearance of bands, increase in thickness and clarity of the bands.

Study of Staphylococcus aureus isolated from the mouth of canary

Saba A. Hussein

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 301-304
DOI: 10.33899/ijvs.2019.125937.1192

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.

Genetic detection to Aeromonas hydrophila proteolytic activity in milk samples (cows, buffaloes and goats) in Basra governorate

Rawa B Banay

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 253-258
DOI: 10.33899/ijvs.2019.125888.1174

This study aim is to determine the incidence and the virulence of Aeromonas hydrophila in raw milk, randomly collected from Basra governorate by using of polymerase chain reaction (PCR) technique. In this study, the total number of raw milk samples collected from cows, buffaloes and goats that kept from different the regions of Basra governorate were 90 samples. The PCR technique is modern method which regarded as a reliable tool to detect virulent gene of the A. hydrophila isolates. The PCR assays using the primers sets SerAh-F and SerAh-R resulted in the amplification of 650-bp bands from the targeted proteases gene of the A. hydrophil. The result of the present study showed that the results of PCR concerning the proteolytic activity of A. hydrophila in the tested raw milk samples according to animals' source. The higher percentage of the proteolytic activity was found in the cow's raw milk samples 40% and in the buffalo's milk samples was 26.7% while, the proteolytic activity did not find in the goat's milk samples. The association between the source of the milk sample and proteolytic A. hydrophila positive results was considered to be statistically highly significant. The higher percentage of the A. hydrophila isolates found in the raw cow milk was 40%, and the A. hydrophila isolates found in the raw buffalo milk was 26.7%, while, the A. hydrophila isolates did not find in the goat milk.

Pathogenesis of Salmonella enterica serovar albany in experimental infected SPF BALB/c Mice

Jesny B. Sabri; Imad I. Al-Sultan; Khalil Altaif; Sherly Peter; Mohammed J. Saadh

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 339-344
DOI: 10.33899/ijvs.2019.126269.1282

Salmonellosis remains an important zoonotic disease and public health concern, Salmonella enterica serovar Albany is one of the motile serovars which has been identified from poultry and humans. However, its pathogenic potentials and shedding probability and duration from infected/colonized chickens have never been reported. To assess its pathogenic potentials and shedding probability 6 SPF BALB/c mice was inoculated with 0.1ml volume for each mice bacterial solution of 108 CFU/ml of Salmonella entrica serovar Albany after 24 hours the segments of the duodenum, jejunum, ileum, caecum and colon were fixed to study the histopathology and the polymerase chain reaction (PCR) was used to confirm the Salmonella entrica serovar Albany in the intestinal mucus swabs. The control group consist of 6 SPF BALB/c mice were inoculated with 0.1ml of 0.9% normal saline, The pathogenesis incidence rate of the disease caused by S. entrica serovar Albany revealed that prominent blood vessels on caecum 100%, red intestinal serosal 100%, infiltration of inflammatory cells in the crypt of liberkhun and submuscular layer of small intestine 100%, enterocyte necrosis 100%, haemorrhagic enteritis 83.3%, cecitis 33.3%, colonitis 66.6%, villus atrophy 100%, crypt atrophy 100%, and detachment of epithelial tissue 50%, can occur as soon as 24 hours post infection. Infected S. entrica serovar Albany was also successfully re-isolated from the intestinal swabs which revealed that the mice is potentially shed the bacteria through feces.

Natural heavy infection with immature sarcocysts of Sarcocytis spp. in sheep in Mosul city: A case report

Nadia S. Alhayali; Manal H. Hasan; Karam Y. Al-Mallah

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 373-376
DOI: 10.33899/ijvs.2019.125994.1210

This study included recording the natural heavy infection with immature macrocysts (Sarcocysts) of Sarcocytis spp. in sheep. The sheep is one years old which is slaughtered at butcher shop at Mosul city in May 2018. This is the first case recorded of natural infection with immature sarcocysts of Sarcocystis spp. in Mosul city. Many of small nodules were observed during slaughter, these nodules are seen within esophageal muscles in different sizes and shapes, they were distributed randomly throughout esophageal muscles. Most of the sarcosystis were small in size the mean of size between 20- 28×28-42 µm they were histological examination showed that presence of only metrocytes. This confirmed the diagnosis that the sarcocysts were immature macrocysts (sarcocysts) for the Sarcosystis spp. In our study, heavy infected case with Sarcocystis reveals the fact that large numbers of cats(final hosts) in contact with sheep in pastures is considered the main risk factor for infection and feed with raw meat from infected sheep, which is very important for carcass condemnation when the meat inspection when abnormalities are found which indicate that the part of carcass, is unfit for human consumption it is condemned, which means the economic loss for livestock.

Inhibition of Escherichia coli biofilm formation by Streptomyces sdLi crude extract

Ahmed J. Neamah; Abdul-kareem S. Al-Al-Yassari; Mohammed A. Hamed; Miran A. AlRammahi

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 2, Pages 305-310
DOI: 10.33899/ijvs.2019.125965.1202

Biofilm is a microbial-protecting environment initiated on surfaces that reveals major health problems such as biofilms represented by dental plaques. Fighting biofilm formation is a hugely demanded process. Here, the crude extract of Streptomyces sdLi (sediment lake Iraq-sdLi) was used to check the anti-biofilm formation bioactivity (ABFB) against Escherichia coli (Orooba Meteab Diwanyah 4, OMD4) isolated from milk samples. Using a cross-streak method, each strain of Streptomyces spp. was tested for the best broad-spectrum ABFB. A triplex polymerase chain reaction (TPCR) method targeted specific genes and a fragment (hemin receptor molecule (chuA), uncharacterized protein YjaA (yjaA), and chuA TspE4.C2) was used to categorize 18 isolated OMD4. Using the alcoholic extract of liquid growth of the best strain with ABFB, a crystal violet biofilm assay (CVBA) was employed to test the ABFB against OMD4. The results of the screening test revealed Streptomyces sdLi with strongest ABFB; however, ethyl acetate, as one of the sdLi extracting solvents, was the most potent in in inhibiting the biofilm formation. The TPCR resulted in 18 isolates categorized into four groups A, B1, B2, and D in which B2 and D are known for their significant pathogenic activities in humans and animals. The results of the CVBA showed that Streptomyces sdLi extract was potential for its ABFB. This study recognizes that the Streptomyces sdLi extract is potential for deactivating biofilm formation by pathogenic E. coli which encourages future studies to consider this microorganism and/or its extract as a cure for the treatment of E. coli related illnesses in humans and animals.

Bioremediation of lead and cadmium and the strive role of Pediococcus pentosaceus probiotic

Raghad Jaafar

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 51-57
DOI: 10.33899/ijvs.2019.125581.1092

Consumption of food and water contaminated with heavy metals poses a huge threat to the life. Both of Lead (Pb) and Cadmium (Cd) are heavy metals and important environmental pollutants. Away from traditional treatments, the current study aims to adopt probiotic bacteria Pediococcus pentosaceu to treat heavy metal pollution. Present results indicated a good probiotic property of P. pentosaceus, where they were able to survive pH range from 3-9, during incubation periods 3 and 24 hours, and bile salt range 0.15-0.5% for the same period. The number of bacteria in gastric (pH 3) and intestinal juices (pH 8) after 24 hours of incubation was 390 and 205, respectively. Bacteria showed an inhibitory effect against pathogenic bacteria Salmonella sp. The antibiotic susceptibility test revealed them resistant to clindamycin, intermediate resistant against benzylpenicillin, ampicillin, and their sensitivity to the rest tested antibiotics. Isolated bacteria identified based on their morphology, biochemical characteristic in addition to the use of automated instrument for bacterial identification (Vitek II), and depending on the results bacteria were identified as P. pentosaceus. In bioremediation study, the lowest inhibitory concentration of lead and cadmium and (MIC) was done, followed by assay the removal capacity by P. pentosaceus, using atomic absorption spectrometry (AAS) analysis. Bacteria show high MIC (1800 and 150 ppm) for Pb and Cd respectively. With removal efficiency for Pb 62.10-68.39% in the concentrations 25 and 50 ppm, respectively, and for Cd 52.71-11.25% in the same concentrations. Depending on the present finding probiotic bacteria (P. pentosaceus) can apply in the bioremediation of heavy metals in the fish ponds when contamination occurs, in addition to their tradition used as safety additive to prevent fish disease and an enhancement agent .Finally the isolation of these bacteria from fish ponds can be considered as a good indicator for a healthy state of fish ponds in the studied area.

Detection of Mycobacterium paratuberculosis in raw cow’s milk using polymerase chain reaction (PCR) technique

Ihsan M. Ahmed; Raad A. Al-Sanjary; Haiffa H. Alkazaly

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 83-86
DOI: 10.33899/ijvs.2019.125556.1075

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.

Detection of methicillin-resistant Staphylococcus aureus in broiler and broilers farm workers in Duhok, Iraq by using conventional and PCR techniques

Mahde S. Assafi; Hishiyar A. Hado; Ibtessam S. Abdulrahman

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 15-22
DOI: 10.33899/ijvs.2019.125757.1145

Livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) has become a global public health concern. The purpose of this study was to investigate the prevalence rates of MRSA infection amongst broiler chickens and broilers farm workers. The total samples used in this study were 306. Cloacal swab samples from 231 broilers and nasal swab samples from 75 broilers farm workers were collected from five farms in Duhok city, Iraq. Isolation and identification of MRSA isolates were carried out and the antibiotic susceptibility were screened. Molecular characterization of all isolates was performed by using polymerase chain reaction (PCR) technique to detect the mecA gene.S. aureus was detected among 84% (63/75) of the farms workers samples and among 84.8% (196/231) of the broiler's samples. The S. aureus isolated from farm workers and broilers appeared resistant to oxacillin 28.6% (18/63), and 32.1% (63/196), respectively. MRSA colonization in farm workers and broilers was 24% (18/75) and 27.3% (63/231) respectively. The S. aureus isolates showed the most resistant to chloramphenicol and the least resistant to vancomycin. The results of the PCR assays revealed that 85.7% (12/14) of S. aureus isolates from farm workers and 44.4% (16/36) of S. aureus isolates from broilers were positive for the mecA gene. The direct handling of broilers by farm workers plays the important role for transport the MRSA isolates from broilers to broilers farm workers.

Investigate the Toxoplasma gondii infection in the consumed beef in Al-Diwaniyah province

Farah M. Sakban; Noman N. A’aiz

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 95-99
DOI: 10.33899/ijvs.2020.164336

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.

Detection of Shigella in raw bovine milk by polymerase chain reaction

Noor Soulieman; Aemaan Al-Mariri; Faizah Al-Atrash

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 9-16
DOI: 10.33899/ijvs.2019.125758.1146

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.

Detection of the extended spectrum β-lactamase produced by Escherichia coli from dairy cows by using the Vitek-2 method in Tulungagung regency, Indonesia

Akyun R. Putra; Mustofa H. Effendi; Setiawan Koesdarto; Suwarno Suwarno; Wiwiek Tyasningsih; Acts T. Estoepangestie

Iraqi Journal of Veterinary Sciences, 2020, Volume 34, Issue 1, Pages 203-207
DOI: 10.33899/ijvs.2019.125707.1134

The use of antibiotics in inappropriate on food producing animals can lead to resistance many of the pathogenic bacteria to the various types of antibiotics, one of which is the Escherichia coli (E. coli) which produces extended spectrum β-lactamase (ESBL). Antibiotic resistance in animals and humans has become a global problem that needs attention and immediate management by using specific antibiotics that used for therapeutic the infected animals. The aim of this study was to isolate and detect E. coli producing ESBL. All E. coli from the surface of dairy cow rectal swabs in Sendang District, Tulungagung Regency, Indonesia using the Vitek-2 method. The number of rectal swab samples used in the present study was 50. The results of this study showed that all the samples were suspected of being E. coli, based on the morphological growth of colonies on the EMBA media. The isolates were identified by using the biochemical tests. All the samples were positive. In this study the double disc synergy test (DDST) method was using to confirm the ESBL. The antibiotics were used amoxicylyn-clavulanate, ceftazidime and cefotaxime for DDST. In additional ESBL confirmation test was used the Vitek-2 method. The presence of ESBL producing by E. coli isolated from rectal dairy swabs in tulungagung was 6% (3/50).

Isolation and identification of Salmonella typhimurium bacteria with detection of type-1 fimbriae coding gene by polymerase chain reaction (PCR) technique

Zeena Saleh; B.M. Al-Muhana; Kh. Hamdan; M.S. Jawad; S.F. Klaif

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 195-199
DOI: 10.33899/ijvs.2019.162961

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.

Different vectors used to transform and clone of nonstructural NS1 gene of Influenza B in Escherichia coli

Ali Dawood

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 329-333
DOI: 10.33899/ijvs.2019.162964

Flu is a highly contagious and common illness caused by influenza A, B, and C viruses. The aim of the present study was to investigate the transformation and cloning of NS1B gene with pET-32a, pET-32b and pQE-81L in Escherichia coli BL21(DE3) and DH5α. pUC57-NS1B synthetic gene was transform and clone in Escherichia coli BL21(DE3). Isolation, single digestion and ligation with pET-32b using HindIII restriction enzyme. Amplification of recombinant DNA was done with conventional PCR after transformation. Screening with IPTG of colonies. Gel electrophoresis was done for each step of cloning after isolation. Isolation, double digestion and ligation with pET-32a and pQE-81L using SacI, PstI and HindIII respectively.Recombinant DNA was attempted to be transformed into E. coli strains BL21 (DE3) and DH5α. pUC57 plasmid carrying NS1B gene was successful transformed and isolated from E. coli BL21 (DE3). Designed primers used for PCR of NS1B showed successful amplification. First screening of pET-32b-NS1B colonies using white/blue method, cloning NS1B into pET-32b using single restriction digestion with HindIII, pET-32a using double restriction digestion with SacI and HindIII and pQE-81L using double restriction digestion with PstI and HindIII gave unexpected result. This result may relate to re-ligation of digested vector for single digestion and uncompleted digestion for vectors of double restriction digestion. Current study has suggested that recombinant NS1B gene can be cloned using single digestion with other expression vectors.

Detection of Mycoplasma gallisepticum and Mycoplasma synoviae by using of cultural and PCR technique

N.A. Jafar; Bashar Noomi

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 469-473
DOI: 10.33899/ijvs.2019.125484.1016

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.

Propagation of Salmonella enterica serovar typhimurium in embryonated chicken egg

Ammar Al-AAlim; Sumaya Aldabbagh; Haiffa Hussen Alkazaly; Mozahim Al-Attar

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 81-86
DOI: 10.33899/ijvs.2019.163171

The study included the injection of Salmonella enterica serovar typhimuriumisolated from starlings bird in embryonated chicken eggs. The eggs were divided into eight groups, each group contain 6 eggs. The G1 and G2 groups were injected with the sterile normal saline solution in the choriaollantoic membrane (CAM) and yolk sac as negative control. The group G3 and G4 injected by bacterial suspension at a concentration of 104 cfu/ml in the CAM and yolk sac while the G5 and G6 injected with bacterial concentration 106 cfu/ml in the CAM and yolk sac respectively. Finally, the G7 and G8 groups were injected with 108 cfu/ml of bacterial suspension in the CAM and yolk sac respectively. The results showed that the highest percentage of death in eggs embryos was 100% in the sixth group after 96 hours of injection. There was also a significant increase in the number of bacteria in correlation with time of incubation. The highest rate of bacterial isolate was 8,19log10, 8,26log10 after 96 and 144 hours in the sixth group, while the highest number of bacterial isolates was 7.04log10 and 6.31log10 in the third and fourth groups after 48 and 96 hours of injection respectively. The results of the statistical analysis showed a significant difference in the number of bacterial isolate after 24 hours of injection in both concentrations compared to other incubation times. A significantrelationship was also found between the amount of the dose used and the bacterial disease. this study concluded that Salmonella enterica serovar typhimurium isolated from starlings can cause pathological changes and effect on hatchery percentage in embryonated chicken eggs.

Evaluation the efficiency of the fungus Metarhizium anisopliae as biocontrol agent for adults of hard ticks Hyalomma anatolicum

S.Sh. Shahatha

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 57-62
DOI: 10.33899/ijvs.2019.163088

This study was conducted to evaluate the efficacy of the fungus Metarhizium anisopliae  as a vital agent for the control of the Hyalomma anatolicum, which is infested on buffalo fields in some villages of Anbar province, Iraq. The results showed that different concentrations of the fungus 4.2*110, 4.2*310, 4.2*510 pg/ml were capable of killing the tick eggs, and the kill rate was proportional to the higher concentrations used. After 3 days of treatment, moreover causing a high proportion of phenotyping deformation in male and female ticks.

Nad1 gene analysis of Echinococcus granulosus from sheep in Aqrah city, Iraq

R.N. Hamoo; nashaat Ghalib Mustafa; S.A. Abdulraheem

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 341-345
DOI: 10.33899/ijvs.2019.162965

Echinococcus granulosus (E. granulosus) is a dog tapeworm cestoda; it is larval stage responsible to cystic echinococcosis, one of the most common and dangerous worldwide zoonotic parasitic disease. The aim of this study was the molecular identification of the local strain of E. granulosus isolated from sheep liver slaughtered in the principal abattoir of Aqrah city, Northern of Iraq during Jun-Nov. 2017. In this study, 37 sheep liver infected by E. granulosus, 12 of high DNA purity fertile (have protoscolices) cyst of them were considered. A molecular study conducted on the mitochondrial NADH dehydrogenase 1 (nad1) gene. Results demonstrated that E. granulosus isolates were sheep strain (G1) genotype, with fascinating highly corresponding 95% and 96% to global isolates, particularly to north African and Mediterranean countries, by employing phylogenetic tree analysis. So, the isolates of our project were deposited in Genbank (accession No. MG792129). This study findings provide that the local isolates of E. granulosus from sheep liver in Aqrah city, Northern of Iraq are loyally equivalent to global strains and isolates, in addition, nad1 gene considers a perfect biomarker in a molecular identification and phylogenetic study of this parasite.

Molecular identification of allelic genotypes of pyrethroid-insecticide resistance in housefly, Iraq

Monyer Abdulameir Abd Alfatlawi; Mansour Ali; Hassan Naser

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 209-212
DOI: 10.33899/ijvs.2019.125538.1060

According to global-wide presence of insecticides resistance to pyrethroids, the current study identified the purpose to detect the allelic genotypes regarding this issue in house flies in Iraq. From the governorate of Al-Qadisiyah, Iraq, 60 morphologically and molecularly recognized house flies were caught from 6 different regions. Using a technique called polymerase chain reaction (PCR) amplification of specific allele (PASA), PCR was employed to reveal the presence of allele-genetic variations in the para-type sodium channel (para) gene to recognize knockdown resistance (kdr) mutation from the homozygous-wild type of complete susceptibility (sus/sus) to the mutated-homozygous type of complete resistance (kdr/kdr) or to the mutated-heterozygous type (kdr/sus). Here, these genotypes were targeted using specific primers to identify these genetic variations. The results have declared the presence of the sus/sus at 100%-frequency rate in all flies, and none of the other genotypes were detected (0%) in all flies. This valued piece of result indicates the reality of resistance persistence due to lack of insecticide-spraying programs in the governorate. This study provides high-quality information about the current status of insecticide resistance in house flies in Iraq about supporting the fact of genetic-base development of such resistance via frequent use of insecticides.

Serotyping, virulence gene expression and phenotypic characterization of E. coli O157:H7 in colibacillosis affecting buffalo calves in Basra governorate

H.A. Naji; Wessam Monther Mohammed Saleh; M. Hanoon; I. Imad; Y. Salim

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 445-451
DOI: 10.33899/ijvs.2019.163198

The objective of the current study was to detect the clinical signs of colibacillosis in buffalo calves, isolate E. coli O157:H7, detect its virulence gene eaeA using PCR and estimate its prevalence.The current study sampled 120 buffalo calves aged 1day to 5 months from the Al- Basra veterinary Hospital and Private veterinary clinic within the Basra province between October 2017 and July 2018. A total of 100 calves were naturally diarrheic and the other 20 calves served as controls. The clinical sings in the diarrheic subjects included a significant increase in body temperature, heart rates, respiration rates and capillary refill time as compared to control group. Other clinical signs included whitish to yellowish watery diarrhea with tincture of blood, anorexia, weakness, depression, weak suck reflex, dry oral mucous membranes, cold extremities, weak peripheral pulse, dehydration and death. Using phenotypic characterization tools like MacConky agar, EMB agar, biochemical tests and Viteck, 83 out 100 diarrhea samples confirmed E. coli. Using CT- SMACT agar, 31 out of 83 E. coli isolates were E. coli O157:H7 positive. The PCR result indicated that 47 out of the 83 isolated E. coli samples were positive for eaeA virulence gene. In conclusion, this study is a debut in the report of E. coli and E. coli O157:H7 isolation and genes identification in buffalo calves in Iraq. Therefore, proper prevention and control measures are requisite to curtail the mortality and morbidity rate caused by Colibacillosis.

Diagnostic study for Listeria monocytogenes isolated from brain and meat of sheep in Mosul city

Sumaya Aldabbagh

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 2, Pages 51-55
DOI: 10.33899/ijvs.2019.163087

This study included collect of 150 samples from brain and meat of sheep from the slaughterhouse and local butchers shop in Mosul city. 50 sample from each (brain, cutting meat, and minced meat) which used for detection of listeria monocytogenes. The International Standard Organization (ISO) methods were used for isolation. The isolated bacteria were diagnosed according to bacterial morphology, culture, and biochemical characteristics. 10 isolates were obtained, which included 2(4%) isolates from the brain of sheep, 3 (6%) isolates from cut meat and 5(10%) from minced meat. Virulence factors tests were used for bacterial isolates which include, lecithinase, lipase, protease, esterase, and hemolysin. Antibiotic sensitivity test for bacterial isolates was also used for some antibiotics. The results indicated that all isolates were sensitive to Ampicillin, Gentamycin, Chloramphenicol, and resistant to Nalidixic acid. However, they showed variant sensitivity to other antibiotics. In conclusion, this study documented that L monocytogenes can be isolated from brain and meat of sheep in Mosul city.

Molecular characterization of enterohemorrhagic E. coli O157 and O153 isolated from tissue camel and human stool samples in Al-Diwaniyah, Iraq

Saba Falah Klaif; Zeena Foaad Saleh; Alaa abd alkadhem Jawad

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 81-86
DOI: 10.33899/ijvs.2019.125530.1052

The present study aimed to describe the genetic relationships of zoonotic characterization of Escherichia coli isolated from human and livestock camel clinical infection. The study includes collected (50) meat of camel and (50) stool human samples. These samples were foreword to traditional bacterial isolation and identification using enrichment culture method and biochemical tests, then confirmed by PCR technique based on Gyr B gene Escherichia coli and DNA sequencing was done on some positive isolates. The results show that Escherichia coli were isolated from animals at 42 (84%) and 39 (78%) from human infection. The PCR technique was show highly sensitive and specific confirmative detection of Escherichia coli the positive results into 40 (95%) meat sample of camel, and 35 (89.7%) stool sample of a human. To evaluate of Virulence E.coli,we used specific virulence hlyA gene from NCBI-GenBank, published sequence of E. coli hly A gene (Genbank code: X94129.1) and the results show high of presence of virulence gene hly A in camel in percentage (19) 45% than of virulence gene in human (15) 38%. DNA sequencing of a partial sequence of GyrB gene was shown highly homology sequence identity with NCBI-Blast Escherichia coli strain O157H7 isolates from human and O153H3 from the camel. The phylogenetic analysis was shown there is clear genetic similarity at between human and animal’s E. coli isolates and then the gene sequence deposited into NCBI-Genbank accession numbers (MG560867.1, MG560866.1). Also, study design for detection of some virulence gene hly A Escherichia coli. In conclusion, there prevalence E. coli in humans and camel. Therefore, it is essential to define the role of animals as an important source for the distribution of pathogen related to public health. Our study found gyrB gene sequence could be used for identification and making a phylogenetic analysis of gyrB nucleotide.

The genetic relationship for Klebsiella pneumoniae isolated from human urinary tract and beef

Saba Falah Klaif; Hassan Naser; Jenan N. Sadeq

Iraqi Journal of Veterinary Sciences, 2019, Volume 33, Issue 1, Pages 75-80
DOI: 10.33899/ijvs.2019.125531.1053

The present study aimed to describe the genetic relationships of zoonotic characterization of Klebsiella pneumoniae isolated from Human urinary tract and beef. The study includes (50) urine samples from human and (50) beef samples. The isolation and identification of Klebsiella pneumonia were done by using enrichment culture method and Vitek 2, then confirmed by PCR technique based on 16S ribosomal RNA gene which designed in this study using NCBI-GenBank (LT599801.1) and DNA sequencing was done on some positive isolates. The results show that Klebsiella pneumoniae was isolated from Beef at 38(76%) And from human at 32(64%) by vitek2. The PCR technique was show highly sensitive and specific confirmative detection of Klebsiella Pneumonia isolates at Clarify DNA sequencing of a partial sequence of 16S ribosomal RNA gene was shown homology sequence identity highly with NCBI-Blast Klebsiella pneumoniae isolates. The phylogenetic analysis was show clear genetic similarity at (0.5 genetic change) between human and beef in Klebsiella pneumoniae isolates. The gene sequence deposited into NCBI-GenBank accession numbers (MF314450.1, MF314451.1, MF314452.1, MF314453.1). In conclusion, the study presents the first report in Iraq of genetic relationship among K. pneumoniae isolates from beef and humans. Therefore, it is essential to define the role of animals as an important source for the distribution of pathogen related to public health.

Plasmid transformation and curing of nalidixic acid gene in Staphylococcus aureus isolated from buffaloes mastitis and workerʼs hands

D. A. Khaleel; R. M. Othman; B. Y. Khudaier

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 2, Pages 167-174
DOI: 10.33899/ijvs.2019.153845

The present study was designed to detect resistant site of nalidixic acid through transformation and plasmid curing of S. aureus strains isolated from buffalo milk with subclinical mastitis and workers’ hands. A total of 37 S. aureus isolates including 17 isolates recovered from buffalo milk infected with subclinical mastitis, in addition to 20 isolates recovered from workers’ hands. All 37 isolates were investigated by detection of the 23S rRNA gene and various other species specific genes including coa, nuc and clfA. The antibiotic resistance of S. aureus isolates was performed by the discs diffusion method using 19 antibiotics. Plasmid transformation method was carried out by transferring the plasmid isolated from S. aureus into competent Escherichia coli HB 101 in order to detection the resistant site of nalidixic acid. Plasmid curing was accomplished by preparing different concentrations of nalidixic acid (100, 150, 200, 250 and 300 µg/ml) and cultured transformed E.coli on LB agar supported with each of the aforementioned concentrations. The molecular results showed that six isolates (five isolates from milk samples and one from workers’ hands) were identified as S. aureus by coa, nuc, and clfA species specific primers. The six S. aureus isolates were found to be resistant to at least 5 antibiotics which included the nalidixic acid. The results of plasmid transformation revealed that E. coli was able to grow on LB agar supported with 100µg/ml, 150 µg/ml, 200 µg/ml and 250 µg/mlof nalidixic acid and failed to grow on 300 µg/ml concentration.

Antimicrobial and antifungal activity of pumpkin (Cucurbita pepo) leaves extracted by four organic solvents and water

H. Mohammed; R.S. Najem; S.S.A. Altekrity

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 1, Pages 33-39
DOI: 10.33899/ijvs.2018.153791

Pumpkin is a rich source of vitamin A, being high in beta-carotene, a precursor to vitamin A. It provides substantial fiber, niacin, and lutein (important antioxidant). Pumpkin seeds have many health benefits, some of which include a good source of protein, zinc, and other vitamins, and are even said to lower cholesterol, Pumpkin plant was mentioned in the holy Quran as protector to protect the prophet Yonah, peace upon him after his expulsion from the whale. The present work was design to elucidate and evaluate different organic solvents i.e. (Distilled water, Ethanol, Hexane, and Petroleum ether) extracts of pumpkin leaves against some of the pathogenic bacteria and fungi. The results showed pumpkin leaves extracts were able to inhibit bacterial (Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, Proteus mirabilis and Pseudomonas aeruginosa) and fungal (Aspergillus fumigatus, Aspergillus niger, and Candida albicans) growth, comparable with the known antibiotic Ciprofloxacin and the antifungal drug Kenazole. There were no significant differences among different solvents in their ability to produce anti- microbial activity except petroleum ether. Petroleum ether extracts did not show any bacterial growth retardation while it showed anti –fungal inhibition in higher concentrations for Aspergillus fumigates and Aspergillus niger, while Candida albicans seem to be resistant to the petroleum ether extract of pumpkin leaves.

Detection of Escherichia coli hlyA gene and Staphylococcus aureus Sea gene in raw milk of buffaloes using RT-PCR technique in AL- Qadisiyah province

J.N. Sadeq; Kh.H. Fahed; H.J. Hassan

Iraqi Journal of Veterinary Sciences, 2018, Volume 32, Issue 1, Pages 87-91
DOI: 10.33899/ijvs.2018.153815

The aim of this study was to determines the prevalence of virulence gene hemolysin A (hly A) Escherichia coli and Staphylococcal enterotoxins (sea) in Staphylococcus aureus in raw milk buffaloes. In molecular laboratory, real-time polymerase chain reaction (RT-PCR) technique has been performed for 24 samples which have been taken randomly from Buffaloes milk, using primers of high specificity for Escherichia coli hlyA gene and Staphylococcus aureus Sea genes. The results showed different degrees of the studied genes activities. Four out of 24 samples represented S. aureus Sea gene (16.6%) whereas 16 out of 24 samples represented E. coli hlyA gene (66.6%). this study concluded that buffaloes milk might be a source of contamination with pathogenic bacteria of virulent genes which may have different levels of activities.

Investigating and identifying Chlamydia psittaci in asymptomatic and symptomatic domestic dogs in middle province of Iran

M.M. Sarmeidani; P. Keihani; P. Rezaei; H. Momtaz; S.H. Heidari

Iraqi Journal of Veterinary Sciences, 2017, Volume 31, Issue 2, Pages 91-94
DOI: 10.33899/ijvs.2017.145603

C. psittaci is one of the dog’s pathogen which can cause respiratory disorders in various hosts and human beings. Chlamydiae are obligatory interacellular bacteria which belong to Chlamydiales. Conjunctival and pharyngeal swabs were taken from 50 captive dogs presented at veterinary clinics of Isfahan and Shahrekord to determine the percentage of infection and prevalence of C. Psittaci in domestic dogs. Samples were collected during 2014 from a total of 7 different breeds of dog; 1-German shepherd 2-Terrier 3- Mixed Poodle 4-Doberman pinscher 5-Persian sheepdog 6- Siberian husky 7-Pekingese breeds were sampled. The molecular PCR method was used to detect this microorganism in captive dogs and C. psittaci was detected in 9 (18%) of them.

Isolation and identification of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli from broiler in Erbil, Iraq

M.N. Al-Sharook; A.A. Hassan

Iraqi Journal of Veterinary Sciences, 2017, Volume 31, Issue 1, Pages 31-38
DOI: 10.33899/ijvs.2017.126717

Extended-spectrum beta-lactamase-producing Escherichia coli isolated from slaughtered broilers in retail market that sell live chickens in Erbil city, Iraq. Forty-one cloacal fecal samples from broiler caecum were investigated from January to April 2016. ESBLs strains were isolated using MacConkey agar supplemented with cefotaxime 1 mg/l and the isolates were identified phynotypically by biochemical tests, TBX agar and VITEK-2 compact system. A total of 34 Escherichia coli and 4 Proteus mirabilis were analysed for determination of ESBL/AmpC by disc diffusion test using antimicrobial 68DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + ESBL inhibitor, cefpodoxime + AmpC inhibitor and cefpodoxime + ESBL inhibitor + AmpC inhibitor and 67DC MAST® ESβL discs group including cefpodoxime, cefpodoxime + clavulanate, ceftazidime, ceftazidime + clavulanate, cefotaxime and cefotaxime + clavulanate. The phenotypic results showed that in group 68DC discs 23.7% E. coli were resistant to cefpodoxime and in group 67DC discs 73.7% of E. coli and 7.9% of P. mirabilis were resistance to one or more of the cefpodoxime, ceftazidime and ceftazidime. Final results revealed that 78.0% of samples were ESBLs/ AmpC positive. This study is the first examination to determine phenorypically E. coli producing ESBLs/AmpC in broiler chickens in Iraq. Conclusion, the healthy broiler can be a major source of ESBLs/AmpC and the possibility that transmitted to humans through the food chain, direct contact and the surrounding environment raises the concerns about public health and safety of poultry meat and the negative consequences of drug therapy that causes the spread of antibiotic resistance.

Evaluation of role of probiotic IMBO reduce the effect of collibacilosis in newborn calves

A.J. Aletabi; J.M. Khalaf; A.S. Albana

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue Suppl. IV, Pages 321-328
DOI: 10.33899/ijvs.2012.168754

The aim of this sutdy is to evaluate the effect of using the (IMBO) as probiotic in the reduction of diarrhea caused by E.coli in newborn calves. Using 150 newborn calves naturally infected with E.coli which is divided into three groups 50 animals in each group. First group, the animals of this group were inoculated Biomin IMBO as probioric orally given daily for 16 day with 6 gm\animal, while second group, the animals of this group were inoculated IMBO as probitic with same previous dose after natural infected with E.coli and suffering from diarrhea. While the third group (control group) which was not inoculated the IMBO as probiotic. The E.coli was isolated and it is virulence was evaluated by k99 pili test, the bacterial count for the feces sample of the calves was done as many hematological test which include (leukocyte count, differential WBCs count, packed cell volume, as well as TPP concentration and albumin and globulin for day (1,7,16) of the study. The probiotic was positively effective in the decrease of the severity of clinical sings and significantly increase calves weight which were treated by the probiotic, in the day (16) of the study and increase in the mortality rate compared with control group. The study shows probiotics use contributed in decrease of the number of E.coli excreted by feces of naturally infected calves with colibacillosis. Hematology test results showed significant increase in total leukocyte count, neutrophil, monocyte count and significant decrease in pact cell volume in the first and second group compared with control group. While serum test showed significant increase in total protein and globulin and significant decrease in albumin value. From the results of the study we notice the positive effect of probiotic IMBO on the health state after causing infection by colibacillosis and decrease the severity of clinical signs and increase the immunity respond.

Hypersensitivity pneumonitis by actinomycetes and it is relation to IgE and IgG levels

R. S. Al-Esayad; E. Gh. H. Al-Sammak

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue Suppl. IV, Pages 329-338
DOI: 10.33899/ijvs.2012.168755

Isolation and identification of the species related to actinomycetes which have a role in Hypersensitvity Pneumonitis (HP). A total of (241) samples were collected from different sources including 83 sputum samples, 83 blood samples from the workers in cows, sheep poultry farms, poultry slaughter house, textuary, flour, tannery factories, sawmills, and hospitalized patient as well as 25 samples from the worker environment (soil, air, water), through August 2008 to February 2009. The isolates were identified to species level depending on morphological, biochemical and physiological tests, including the species Nocardia spp, Rhodococcus equi, Saccharomonospora viridis, Nocardia nova, Coryneformbacteria were isolated in high percentage from individual with normal level of IgE 100 IU\ml with and without clinical symptoms, and with less frequency the species Nocardia spp, Citrococcus spp, Coryneformbacteria in individual with high level of IgE > 100 IU \ ml. The species Saccharomonospora viridis, Nocardia nova, Nocardia spp. also were isolated from individual with high level of IgG.> 25 IU \ ml compared with whom have normal level of IgG 25 IU \ ml. were the species Saccharomonospora viridis, Coryneformbacteria, Nocardia spp, Citrococcus spp, Rhodococcus equi, isolated. But not isolated from the control.

Relationshipe between lipid peroxidation and parasitic infection in birds

A.A. Ashour; A.I. Ahmad

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue Suppl. III, Pages 303-307
DOI: 10.33899/ijvs.2012.168741

The present work was concerned with an effect of the different parasitic infection in pigeon on some importance biochemical changes in these birds such as malondialdehyde level (the last produce of lipid peroxidation) and cholesterol level. Stale these mode of studies are limited. This study include two major steps, the first stage includes diagnosis many types of parasitic infections in the pigeon body, which divided in three main groups; Blood parasitic infection, Gastro intestinal parasitic infections, and Mixed parasitic infections. After that the study was designed to investigate the role of biochemical changes (Lipid peroxidation and cholesterol) associated with parasitic infection in pigeons. Results of the present study show that Many parasites were diagnosed like Haemoproteus sp, plasmodium sp, Eimeria sp, Ascaridia sp, and Subulura sp. Healthy pigeon showed a significant increase in serum malondialdehyde when compared with the parasitic infected pigeon, in three types of parasitic infection, however blood, gastrointestinal and mixed infection. In comparative between the types or three group of parasites in pigeons, we saw that significant increasing in malondialdehyde level in blood parasitic infection compared with gastrointestinal infected, while there's no significant change between gastrointestinal infected and mixed parasitic infected, also there's no significant changeable between blood infection and mixed parasitic infection. The cholesterol level in serum of pigeon didn't show a significant change between healthy and infected pigeon.

Detection of organ bacterial load in quails

Mohammad Ali Hamad; Ammar M. Al-AAlim; Sumaya Y. Aldabbagh; H.H. Ali

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue Suppl. II, Pages 47-51
DOI: 10.33899/ijvs.2012.168711

A total of thirty normally quail birds were bought from local market in Mosul city in the period from September-October 2011. Quail birds were slaughtered and samples taken aseptically from internal organs of each bird for bacteriological investigation. The result showed isolation of 203 bacterial isolates from different organs of quail birds. The isolates ranged from Corynebacterium spp. 29.6 % (60 isolates) as a high percent, then E. coli 18.2% (37 isolates), Staphylococcus aureus 16.3 %(33 isolates), Bacillus spp. 14.8% (30 isolates), Enterococcus faecalis 9.9% (20 isolates), Klebsiella pneumoniae 6.4% (13 isolates), Proteus spp. 1.9% (4 isolates), Pasteurella multocida 1.9% (4 isolates) and Coagulase -vestaphyloco-ccus 1% (2 isolates). This study showedCorynebacterium spp. and E. coli were dominant bacteria in the internal organs of quail birds. Many studies reported that quail birdswere resistant to many bacterial diseases, so that these birds may act asmechanical transporting for different bacterial species to humans and animals with the risky of transporting of resistance bacterial species for many antibiotics.

Detection of Toxoplasma gondii antibodies in horses in Mosul, Iraq

Mohammad N. Alshahery; Reem S. Mansour

Iraqi Journal of Veterinary Sciences, 2012, Volume 26, Issue Suppl. II, Pages 39-41
DOI: 10.33899/ijvs.2012.168677

This study was aimed to verify the presence of Toxoplasma gondii antibodies in equine sera in Mosul city, Iraq. Seventy nine samples of sera were examined (70 female and 9 male) by latex agglutination test (LAT) and 2–Mercaptoethanol test (2-ME). Results showed that anti-bodies to T. gondii using LAT were detected in 72.2% (71.4% female and 77.8% male) whereas 57% (57.1% female and 55.6% male) of infected horses were detected by 2-ME. 

Pathological findings associated with experimental Mycobaterium bovis infection in rabbits

Khalel H. Al-Joboury

Iraqi Journal of Veterinary Sciences, 2005, Volume 19, Issue 1, Pages 83-89
DOI: 10.33899/ijvs.2005.37424

During the 2nd week post inoculation of thirteen rabbits with Mycobacterium bovis tuberculosis lesions appeared in the lungs, liver, spleen, kidney, mediastinal and hepatic lymph nodes and in the omentum with an equal distribution in these organs. During the 4th week post inoculation, these tuberculosis lesions increased in size to become well developed granulomas with caseated centers. These granulomas persisted to the 6th, 8th and 10th weeks post inoculation and became more encapsulated later on. Three rabbits died during the 7th week post inoculation due to generalized tuberculosis.