جستجوی مقالات مرتبط با کلیدواژه « ژن n » در نشریات گروه « دامپزشکی »

Brucellosis or malt fever (Mediterranean fever) occurs due to the growth and multiplication of the gram-negative coccobacillus Brucella in mammalian cells, and this disease is important as a common disease between humans and animals. Several serological tests have been investigated to identify this bacterium, however, these methods have limitations in the accurate identification of the bacterium. The present study was conducted with the aim of investigating and molecular identification of Brucella bacteria in samples collected from infected sheep in Sistan region using multiplex PCR technique. At first, blood and milk samples were collected from infected sheep. Confirmation of bacterial infection was initially done using blood samples and Rose Bengal serology method. Next, DNA was extracted from the collected milk and molecular analysis was performed using multiplex PCR and amplification of Omp31 and BLS genes, in order to amplify fragments with sizes of 347 bp and 256 bp respectively. Out of the 40 clinical samples, all of them showed a positive Rose Bengal test, but molecular analysis using multiplex PCR showed 5 false positive samples confirmed with the Rose Bengal test. Due to the higher sensitivity and accuracy and short time of identification the molecular methods can be used as a suitable option for identifying pathogens.

Gallinarum disease caused by Salmonella gallinarum is one of the most challenging diseases in the poultry industry. In this study, the level of Salmonella gallinarum contamination of industrial chicken flocks in northwestern Iran, antibiotic resistance and frequency of some virulence genes (msgA, spiA, pagC) were investigated and evaluated. All the samples taken from the herds with suspected clinical symptoms of salmonellosis from the clinics of East Azarbaijan, West Azarbaijan, Ardabil and Zanjan provinces were examined and confirmed by microbial culture, chemical analysis, PCR and serological methods. After confirming the diagnosis of involvement with Salmonella gallinarum, antibiotic resistance was evaluated by disking method and the abundance of virulence genes in each isolate was checked. Of the 25 isolates of Salmonella gallinarum in this study, the highest microbial resistance was reported to tetracycline (76%), oxytetracycline (72%) and the highest sensitivity to amikacin (80%) and Fozbek (72%). 100% of the isolates were resistant to at least 6 types of antibiotics. In the study of the isolates, 8% had the msgA gene, 28% had the spiA gene, and 8% had the pagC gene, of which two isolates had all three virulence genes.

Enzymes play a very important role in various industries and in medicine. The BSFA genomic region is also considered one of the most important operons in biotechnology and bioengineering and is used in the preparation of fibrin-breaking drugs (fibrinolytic) or clot-breaking drugs to destroy blood clots in vessels (thrombosis and heart attack treatment) and this enzyme can be used in therapeutic cases. The thermophilic bacillus carries various enzyme genes and the goal is to investigate the genomics and cloning of the fibrinolytic gene (bsfA) from soil thermophilic bacilli in Escherichia coli origami bacteria by Real time PCR and SDS PAGE methods.

Bacillus strains were isolated and identified from a total of 70 soil samples from the areas around Tehran. Then bsfA gene was extracted from bacilli by PCR method. The amplified fragment was inserted into the pTG19 expression vector by TA cloning (Transfer activity) method. In the next step, the recombinant vector was transformed into Escherichia coli origami bacteria and cloning was confirmed using common methods. PCR reaction was performed for the bsfA gene with the mentioned primers.

As a result, all of the 12 Bacillus strains isolated (100%) had the bsfA gene. The results of RNA cloning of suspected colonies extracted cDNA was made and the expression level of bsfA gene was checked by real time PCR test. Determining the molecular identity of the Bacillus genus carrying the bsfA gene was done using primers. Finally, the expression of the bsfA gene in Escherichia coli origami bacteria was confirmed by the sequence of the PCR product.

As a result of this research, they managed to find native bacilli producing bsfA, and finally, successfully transferred the gene of this enzyme from Bacillus bacteria to E.coli bacteria so that this enzyme can be produced in E.coli with more production and economic efficiency. To create bacteria with high expression and recombination ability, it can be a big step in increasing the production of this enzyme in the treatment of patients suffering from thrombosis, which prevents embolism and death.

Enterococci are a part of the opportunistic pathogens, which are very important in medicine. These bacteria can cause a variety of diseases in both dogs and human. The aim of the present study was to investigate the frequency detection of Enterococcus faecalis and Enterococcus faecium in companion dogs in Ahvaz and review of risk factors including age, gender, breed and diarrheic status in animals. Also, the prevalence of virulence genes was evaluated including gelatinase (gelE) and (ccf) and antibiotic susceptibility measured in obtained samples. Sampling was performed from the rectum of the 150 dogs (36 cases diarrheic and 114 non-diarrheic). The samples were evaluated by two methods of bacterial culture and PCR. In bacterial culture, 122 isolates, suspected to Enterococcus species, were isolated and subsequently the detection of SodA gene specific to Enterococcus faecalis and Enterococcus faecium was performed by PCR. Overall forty-five positive isolates were identified, which thirty four of which were Enterococcus faecalis (75.5%) and 11 were Enterococcus faecium (24.5 %). In regard to identify virulence genes (gelE and ccf), 36 out of 45 isolates were positive for virulence genes. Twenty six isolates (57.77%) had virulence genes, 5 isolates (11.11%) ccf gene and 5 other isolates (11.11%) gelE gene. In all, nine isolates (20%) had no virulence gene. Fourteen different antibiotics were used to determine antibiotic susceptibility that indicated all isolates were resistant to azithromycin, streptomycin, ampicillin and imipenem. Thereafter, the highest resistance was related to erythromycin and cephalexin (95.5%), trimethoprim sulfamethoxazole (84.4%) and gentamicin (80%), respectively. Also, the highest sensitivity was related to nitrofurantoin (62.2%), penicillin G (60%) and enrofloxacin (55.5%), respectively. There was no significant relationship between risk factors such as age, gender, breed and diarrheic condition with the presence of Enterococcus in the studied dogs (P>0.05). The results showed that the prevalence of Enterococci was relatively significant (30%) in dogs of Ahvaz district. Antibiotic resistance was significant in the two species of Enterococci. Finally, because of the very high importance of antibiotic resistance, appropriate administration of antibiotics is recommended.

Pets such as dogs and cats are potential sources of transmissible infections such as Yersinia enterocolitica to humans, especially children.The gastrointestinal tract of these animals keeps the infection in them and can be considered as a carrier for their owners. This study was conducted with the aim of investigating the frequency of Yersinia enterocolitica infection in apparently healthy dogs and cats and their owners in Isfahan city. For this purpose, fecal swabs were collected from 115 apparently healthy dogs and cats and their owners, and Yersinia contamination and the presence of virulence genes were investigated using culture and molecular methods. The results obtained in the present study showed Yersinia enterocolitica infection in 32% of dogs, 4% of cats, 24% of dog owners and 4% of cat owners. A significant relationship was observed between age, disease and the way of keeping with the prevalence of Yersinia enterocolitica in dogs, and the level of infection of dog and cat owners was directly related to the infection of dogs and cats. Antibiogram results showed that the highest sensitivity and resistance were related to tetracycline and ciprofloxacin antibiotics, respectively. Infected animals may be the main source of infection. These results can be useful in prevention and control programs.

Today, the widespread use of petroleum products has led to environmental pollution and has created serious problems for the health of the environment. Streptomyces make up 50% of the total population of soil actinomycetes are filamentous, gram-positive, and essentially aerobic. These bacteria are abundant in natural aquatic and terrestrial environments, are not nutritionally hardy, require only a source of carbon and nitrogen along with mineral salts, and they are able to remain in the soil as spores for a long time. The aim of the study was to clone the Streptomyces  catechol 2, 3-dioxygenase gene (C2,3) gene present in the soil in Escherichia coli bacteria in order to remove oil pollutants.

In this study, 58 samples were collected from a depth of 5 to 10 cm in different areas of the suburb of Tehran and oil contaminated areas such as the oil depot and its surroundings. After culturing, diluting and selecting suspicious Streptomyces colonies for identification, after confirmation of Streptomyces bacteria based on colony appearance, microscopic characteristics, and biochemical tests for final confirmation of molecular identification, their 16srRNA sequence was amplified and replicated, was examined. In the next step, after DNA extraction of the samples, PCR was used to amplify the C2,3 gene. The C2,3 gene was then cloned into Escherichia coli Origami by PTG19-T vector. Finally, the amount of gene expression was determined by Real Time PCR and after sequencing the phylogenetic tree was drawn.

The enzyme catechol 2, 3-dioxygenase can be extracted from limited sources such as Streptomyces, so the need for gene amplification of this enzyme is important from the bacterial sources that produce it. Therefore, in this study, this enzyme was isolated from Streptomyces. As a result of this study, we were able to find native Streptomyces that produce the enzyme catechol 2, 3-dioxygenase, and finally, the gene of this enzyme was successfully introduced into Escherichia coli Origami.

In this study, in order to create of a recombinant host with high expression capacity, it can be considered a big step towards increasing the production of this enzyme in the industry. Further investigation to find new strains producing catechol 2, 3-dioxygenase can lead to a new method of removing oil contaminants.

The presence of fat in the carcasses of broilers is one of the main challenges in the poultry industry. Hepatic lipogenesis is an important step related to lipid metabolism in broilers that is controlled by a set of genes. The aim of the present study was to investigate the gene expression profile to identify genes that are effective in the lipogenesis pathway. In the first, the gene expression profiles related to three tissues of the liver, breast, and abdominal fat of broilers fed with two diets containing high starch, low fiber, low fat (LF), and low starch, high fiber, high fat (HF) were downloaded from GEO database. In the next step gene expression analysis related to the three tissue mentioned was performed by GEO2R web-based software. The study of biological pathways was performed by the WebGestalt tool and the gene network was reconstructed by Cytoscape software.  The analysis of the results showed that there was no significant difference in the expression profile due to feeding with two different diets in the breast muscle tissue and abdominal fat, but 975 genes were significantly different in the liver tissue. Based on a network and biological analysis, 10 genes with the highest degree were involved in the biological activities of fatty acid biosynthesis, acyl-coenzyme A biosynthesis, fatty acid metabolism, and fatty acid degradation. Therefore, these genes can be important as key genes controlling hepatic lipogenesis. Therefore, based on our findings, which are the result of the interaction between nutrition and the genome, we can suggest diets for breeding poultry, which can be used as a suitable diet for the production of low-fat meat.

Newcastle disease (ND) is a highly contagious disease in poultry caused by the Newcastle disease virus (NDV). ND is considered one of the most important poultry diseases worldwide and may cause up to 100% mortality in infected chickens. Given the high mortality caused by NDV, the disease diagnosis and characterization of the circulating viral strains is critical. In this study, further studies have been performed to determine the molecular identity of a velogenic Newcastle disease virus genotype XIII.2.1 named RT30/2010, which is the first report of an XIII.2.1 NDV from Iran. In this study, virus purification was performed in two steps by the plaque purification method. The complete coding sequence of the NP gene was investigated to provide additional information about the nature and molecular epidemiology of RT30/2010. For this purpose, the NP gene of this virus was amplified using specific primers and cloned into PTZ57R/T vector. Rapid amplification of cDNA ends (RACE) was used to characterize the 3’ end of the virus genome, which includes the beginning region of the NP gene. Phylogenetic and genetic distance analysis of the nucleoprotein gene of RT30/2010 place the virus in a clade of Pakistani viruses of XIII.2.1. Therefore, RT30/2010 has a different origin compared to other viruses identified from Iran and is in the category of Newcastle disease viruses detected from Pakistan. The information from this study helps to further characterize the virus and makes it possible to use this virus as a challenge or a vaccine strain against XIII.2.1 viruses.

Staphylococcus aureus is one of the most important agents causing mastitis in dairy cattle. The emergence of methicillin resistance in S. aureus is a concern in veterinary medicine and public health. Due to the identification of new types of genes encoding methicillin resistance in recent years, the aim of this study was to investigate the presence of methicillin resistant (MRSA) and types of mec gene for the first time in the northeast of Iran. In the present study, the identity of 54 staphylococcus aureus isolates causing bovine subclinical mastitis were investigated using biochemical and molecular tests. Among them, the species-specific nuc gene was detected in 44 strains. Then, resistance of isolates to penicillin, cefoxitin, enrofloxacin, tylosin and trimethoprim-sulfamethoxazole were determined using the disk diffusion method. Finally, the molecular investigations for detection of the mecA, mecC, mecB genes were conducted. According to the results, the highest resistance were related to penicillin (54%) and trimethoprim-sulfamethoxazole (23%), and the lowest resistance was related to taylosin (7%). The mecA gene was observed in only one isolate (2.3%), while the isolate was not identified as MRSA in phenotypic test. The results of the present study show that the incidence of methicillin resistance in bovine mastitis strains of the area is not high. This study also showed that less known types of mec genes were not common in this region. However, further studies must be perform in this regard.

Pseudomonas aeruginosa is a Rod-shaped Gram-negative and strict aerobic bacterium that can causes infection in the urinary system, respiratory system, middle membrane of the skin, soft tissues, bacteremia, etc. Ceftolozan-tazobactam (C/T) is an effective antibiotic in controlling of this bacterium. But recently, reports of the emergence of resistant strains have created a serious challenge in the fight against infections caused by P. aeruginosa. ampC is one of the main genes for β-lactamase synthesis. Studies have shown that Ceftolozane-tazobactam has better outer membrane permeability and improved stability against chromosomal ampC β-lactamase than do other β-lactam antibiotics. Considering the importance of ampC in the process of P. aeruginosa resistance to the Ceftolozane-tazobactam antibiotic, the co-expression network of ampC was reconstructed and the resistance emerging process was analyzed. 31 genes in the beta-lactamase process in P. aeruginosa showed correlation with ampC. Network topology analyzes showed that five genes ampC, ampD, poxB, ampR and nagZ have the highest importance in the network. ampC gene had the highest number of direct connections with other genes in the network with 16 connections. Eight pathways had the highest number of representatives among genes. Based on the statistical analysis of the significant pathways among the studied genes, it seems there is a direct relationship between pyruvate metabolism and resistance to β-lactam antibiotics. Results showed ampC is the main gene for resistance to Ceftolozane-tazobactam. Although studies have shown that poxA and poxB genes do not have a significant effect on P. aeruginosa resistance to the antibiotics, the importance of these two genes in the reconstructed network emphasizes that more studies should be conducted on them.

Red blood cells play an essential role in the oxygen transport and the immune system. Moreover, hematologic parameters are an important clinical indicator of various diseases including anemia and metabolic syndrome.

The present study aimed to conduct genome-wide association studies (GWAS) based on gene-set enrichment analysis to identify the loci associated with hematological traits using 630K arrays.

For this purpose, the phenotype records included 498 genotyped Alpine Merino sheep were used for red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and RBC volume distribution width coefficient of variation (RWD_CV). Genome-wide association study was performed with hematological traits using TASSEL software. Using biomaRt2 R package R, SNP was assigned to genes. GO, KEGG, DAVID, and PANTHER databases were used to assign the genes to functional categories.

11 SNP markers on chromosomes 1, 2, 3, 6, 8, 10, 11, 14, and 20 located in TRPC4, SPAT1, TMCC2 (RBC), KRT26, GPLD1, EPAS1 (HGB), RAC2, HSPD1, PDGFRA (HCT) and BBS1, HAG1, PIK3R3, STXBP5, FCER1G  (MCH, MHCH, RWD_CV) genes were identified. Based on the pathway analysis, 17 pathways from gene ontology and biological pathways were associated with hematological traits (P˂0.05). The pathways have important functions in the development and differential of red blood cells, hypoxia, adaptation process, environmental stress, and platelet activation.

In total, this study supported previous results from the GWAS of hematological traits, and also revealed additional regions in the sheep genome associated with important traits, using these findings could be potentially useful for genetic selection in the breeding programs.

Salmonella is one of the leading causes of foodborne illness and serious health concerns worldwide. The high Prevalence of Salmonella is one of the problems in the livestock industry. Due to the prevalence of resistance to antibiotics, in this study we evaluated the antibacterial effects of Yarrow hydroalcoholic extract on Salmonella with antibiotic resistance gene isolated from diarrhea dogs. For this purpose, fecal samples of 83 dogs with diarrhea were immediately cultured by swap in differential media. After confirmation of Salmonella isolates in both molecular and microbial methods, the presence of aminoglycoside resistance genes (aac (3) -IIa and aac (3) -Ia) was investigated. The antibacterial effect of yarrow extract and MIC (minimum inhibitory concentration) was investigated. Finally, the results were analyzed in SPSS 20 software. The results of the present study showed that the prevalence of Salmonella bacteria in the fecal sample of diarrhea dogs was 8.5%.In the MIC study among the measured groups, the lowest inhibitory concentration in mg/ml for strains with aac (3) -Ia, aac (3) -IIa, aac (3) -Ia + aac (3) -IIa and isolates without resistance genes were 78.2±66.3, 46.9±22.1, 62.5 and 93.8±44.2, respectively. No significant differences were observed between these groups (p>0.05). Due to the fact that yarrow has different compounds that can have a strong effect on the inhibition of various microorganisms. Therefore, choosing the right treatment strategies such as the use of medicinal plants like yarrow can play an effective role in reducing the prevalence and epidemic of bacterial diseases.

Marek's disease (MD) is a highly contagious neoplastic disease in poultry. The causative virus is an alphaherpesvirus that causes immunosuppression and predisposes poultry to other pathogens.  Although the MD is more common in laying, the incidence of the disease is also increasing in broilers. Due to the increasing rate of infection with the marker virus in broiler chickens, this study aimed to investigate the occurrence of MD in broiler poultry farms in Semnan, Iran. The feather follicle epithelium and spleen tissues were collected randomly from 15 broilers flocks at 6-10 weeks old in Semnan slaughterhouse and were subjected to polymerase chain reaction by amplifying the meq gene. The results of this study showed of 15 flocks, two (13.6%) had a positive reaction. There was no evidence of macroscopic lesions among the studied flocks. Considering the infection of 13.6% of the studied flocks, without any macroscopic complications, it seems the MD virus is circulating in broilers and could transmit to the other birds. It is indicated that the current national vaccination has not stopped the disease in commercial chickens. Due to the immunosuppressive properties of the virus, it is considered that the MD virus, along with other immunosuppressive agents such as adenoviruses, has an essential role in the development of immunosuppression syndrome in broilers in Iran, and to reduce the economic losses caused by this disease, effective prophylactic measures should be considered by the country's health officials.

Salmonella unlike other members of the Enterobacteriaceae family are voluntary intracellular parasites and all of them are potentially pathogenic. These bacteria are easily transmitted directly or indirectly from animal, animal to human, and human to human. Therefore, the prevention and control of salmonellosis in humans are largely dependent on its prevention and control in animals. This study was performed to investigate the molecular properties of Salmonella enteritidis isolates in red meat offered to the consumer market of Shahrekord city. For this purpose, 300 red meat samples were collected135 samples were positive for Salmonella species. In biochemical and molecular studies, 37 isolates of Salmonella enteritidis were detected. Out of 37 Salmonella enteritidis isolates, inv A and stn genes were estimated in all isolates (100%), spvA in 28 isolates (75.67%), and sefA gene in 20 isolates (54%). According to the results of periodic microbial monitoring and tests and more measures by the officials of the provincial veterinary department in order to reduce the amount of microbial contamination is very important.

Pseudomonas aeruginosa a potential risk factor for food poisoning especially in aquatic foods like shrimp. In order to determine and compare the rate of infection in different species of shrimp in two seasons of summer and autumn in Isfahan and Chabahar cities, the abdominal muscle of 35 shrimps in each season of each city were sampled. Samples were homogenized in peptone water medium, then incubated and cultured in PCA medium. Suspected colonies were isolated and then confirmatory tests of Pseudomonas aeruginosa and confirmation of the presence of nanI, the specific gene of Pseudomonas aeruginosa by PCR.In summer, in Isfahan and Chabahar, 48.5% and 25.7% were infected respectively. In autumn, the prevalence was 22.9% for Isfahan and 20% for Chabahar, respectively. In Isfahan, the highest rate of infection was related to Metapenaueus affinis and the lowest to Penaeus semisulcatus in summer. In autumn, the highest number of infected samples was related to Penaeus merguiensis and the lowest to Penaeus semisulcatus. In Chabahar city, in both seasons, the most infected samples were related to Metapenaeus affinis, and the lowest to Panaeus semisulcatus and in autumn was related to both Penaeus semisulcatus and Litopenaeus vannamei.There was a high prevalence of this bacterium in samples of different shrimp species in both seasons and both cities, which indicates a high risk of food poisoning of Pseudomonas aeruginosa due to improper consumption of shrimp. Cold chain observance during storage, transportation and sell of shrimp will be very effective in controlling this problem.

Aspergillosis is an opportunistic fungal infection in animals. The disease is mainly respiratory, but other disease manifestations also occur in poultry. Researchers have shown that, one of the reasons for the increase in drug resistance in Aspergillus species is the overexpression of the CYP51A gene. This study compared CYP51A gene expression of Aspergillus fumigatus isolated from poultry against fluconazole and nano-fluconazole. Fluconazole liposomal-nanoparticles were prepared by the thin layer hydration method. We dissolved 5.12 mg of fluconazole powder in 1 ml of sterile distilled water and 6 ml of chloroform-methanol organic solvent. We added 51 mg of lecithin and 5 mg of cholesterol to it. The size of nanoparticles was 88.9±12.1 nm and the surface charge of these nanoparticles was -20.12±1.88 mv. We also used a Scanning-Electron Microscope to study the structure of nanoparticles. Thirty samples of A. fumigatus were collected from poultry lung nodules. Minimum inhibitory concentration (MIC) was performed by standard Broth Microdilution method according to NCCLS-M38A2 to evaluate the MIC of fluconazole and nano-fluconazole against isolates. We selected two high-resistance isolates to fluconazole and used them to determine the CYP51A gene expression level by real-time PCR. The results showed that nano-fluconazole had a lower MIC than fluconazole and in lower concentrations of the drug inhibited the growth of Aspergillus fumigatus isolates. CYP51A gene expression was increased in fluconazole and nano-fluconazole-treated isolates compared to the untreated state. Conversely, a decrease in CYP51A gene expression was observed in the exposure to nano-drug compared with the normal drug.