Iraqi Journal of Veterinary Sciences

The Escherichia coli (E. coli) bacteria belong to the Enterobacteriaceae family can cause wide ranges of infections including enteric, extra enteric or opportunistic infection especially in young animals. E. coli outer membrane composed of phospholipid and lipopolysaccharide which reflect the inner and outer layer of the membrane, the main components of lipopolysaccharide LPS are lipid A, core oligosaccharide, and O- antigen. Immunity of LPS was clearly understood in the last 20 years when discovered signaling response in immune cell system, LPS acting as potent activators of immune system especially innate immunity and as a result produce a large variety of the adverse, unfavorable physiological response leading to injury of tissue and endotoxic shock and lastly death.
In this study, we tried to extract and purify the LPS from E.coli local strain and determine the molecular weight, purity, and pyrogenic effect of the product and compare with stander E.coli O55:B5 LPS, the result shows that the hot phenol method with enzymatic treatment give highly pure LPS with high yield reach 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 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 method of extraction used in this research was valid and give highly pure LPS with enduring substantial pyrogenic activity.

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

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

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.

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.

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.

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.

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.

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.

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.

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.

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.

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

This study 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.

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.

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.

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.

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).

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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 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.

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.

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.

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.

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.

This updated review tabulates confirmed pathogens of some diseases in selected animal species in Iraq. Laboratory confirmations were done on all the reported cases.

During the 2^nd 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 4^th week post inoculation, these tuberculosis lesions increased in size to become well developed granulomas with caseated centers. These granulomas persisted to the 6^th, 8^th and 10^th weeks post inoculation and became more encapsulated later on. Three rabbits died during the 7^th week post inoculation due to generalized tuberculosis.