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BACKGROUND: Heparin is a sulfated glycosaminoglycan Blood is a common source for DNA detection in all kinds of samples, and anticoagulants such as heparin and ethylenediaminetetraacetic acid (EDTA) are used to prevent coagulation. Because heparin has a strong inhibitory effect on polymerase chain reaction (PCR), it is not used in samples that will be tracked by DNA. There are physical, chemical, and enzymatic methods to eliminate the inhibitory effect of heparin on PCR test. OBJECTIVES: First, to compare the intensity of the inhibitory effect of two anticoagulants, heparin, and EDTA, on the Real-Time PCR (qPCR), and then to investigate the impact of the heparinase enzyme present in the medium culture extract of Pseudomonas aeruginosa, on removing the inhibitory effect of heparin during the real-time PCR. METHODS: In the present study, two blood samples containing heparin and EDTA were subiected to a real-time PCR test to check the intensity of the inhibitory effect. Then, the medium culture extract of Pseudomonas aeruginosa was added to the heparinized blood sample infected with Escherichia coli bacteria in two groups with different conditions. In the first group, the DNA in the heparinized blood sample was extracted by the phenol-chloroform isoamyl alcohol method. Then, these samples were incubated with the extract of Pseudomonas aeruginosa bacteria culture medium at different hours, but in the second group, the samples were incubated at different hours before DNA extraction. Also, the DNA concentration in both groups was measured by a Nanodrop device, and finally, all samples were subiected to a real-time PCR test. RESULTS: The results of the research samples showed that although the heparinized blood sample contains more DNA concentration than the EDTA blood sample, it completely prevents genome replication. Also, incubating heparinized blood with Pseudomonas aeruginosa culture medium extract before DNA extraction for more than 24 hours removes the inhibitory effect of heparin during the real-time PCR, even at a lower cycle threshold than the EDTA-containing sample. CONCLUSIONS: The Pseudomonas aeruginosa culture medium extract may enable researchers to use heparinized blood samples for genome amplification and diagnosis without using expensive and limited commercial heparinase enzyme. [ABSTRACT FROM AUTHOR]

Background and Aim: Anaplasmosis caused by Anaplasma spp. is an important arthropod-borne disease of various vertebrates with health concerns to humans. The present study aimed to investigate the detection and genotype of Anaplasma phagocytophilum using microscopic examination, real-time PCR technique, and phylogenetic analysis in dogs from Mashhad shelters, Khorasan-Razavi province. Materials and Methods: For this purpose, 250 blood specimens were collected during routine health checkups from dogs in different shelters in Mashhad, Iran, in 2020. First, smears were prepared from the blood specimens, stained with Giemsa, and examined under the light microscope for Anaplasma inclusions. Then, the genomic DNAs were extracted from buffy coats of blood specimens and screened by real-time PCR for the presence of Anaplasma infection by amplifying a 1400 bp sequence of 16S rRNA belonging to the Anaplasma genus. Finally, sequencing and BLAST analyses were carried out on the amplified fragments for the phylogenetic assessments in positive specimens. Results: A total of 9 dogs (3.60%), including 5 females (3.40%) and 4 males (3.88%) were found to be positive for Anaplasma infection in real-time PCR. Moreover, in blood smear observation, A. phagocytophilum morulae were detected in the neutrophils of 3 PCR positive animals. Conclusion: This study provides important data regarding A. phagocytophilum in dogs and the degree of genetic homology/heterogeneities among these pathogen strains from dogs and humans in Iran and other countries. To our knowledge, this is the first molecular evidence on the infection of A. phagocytophilum in sheltered dogs of the region. [ABSTRACT FROM AUTHOR]

Infectious laryngotracheitis (ILT) is a highly contagious, infectious respiratory disease that is caused by Gallid Herpesvirus-1 (GaHV-1, namely ILTV) mainly affecting pullets and laying hens. The disease has not been reported in broilers and backyard chicken in Iran, previously. The present study aimed to identify ILTV strains in outbreaks with severe respiratory clinical signs that occurred in Semnan province (Iran) in broiler farms and backyard chicken farms during June 2019 and March 2020. For this purpose, specimens were investigated from twentyeight broiler farms with severe respiratory symptoms and a mortality rate of 5.5-29% and 2 backyard flocks. The presence of ILTV was assessed by histopathology, conventional PCR targeting of ICP4 gene fragment, and sequencing analysis. Comparison of detected genotypes with strains from Iran and neighboring countries demonstrated the high homology of detected isolated with field strains from Turkey. [ABSTRACT FROM AUTHOR]

Background: West Nile Virus (WNV) is an arboviral infection continuing to be as major threat to human health as well as the livestock industry all around the world. Birds including pigeons are one of the potential reservoirs for WNV. This study aimed to detect the presence of WNV genome in feral pigeons circulating in Semnan and Khorasane-Razavi Provinces (Iran) including 10 urban and 10 suburban areas. Methods: Totally, 150 samples (brain and kidney) were collected equally from feral pigeons and the presence of WNV genome was evaluated in these samples after RNA extraction. Results: All the samples were negative for the presence of WNV-RNA in this investigation. Conclusion: Although obtained result indicated no evidence of WNV genome in feral pigeons but complementary studies regarding serologic detection of WNV in vertebrate hosts as well as pigeons and identification of arthropod vectors seems necessary for comprehensive determination about infection status in these areas. [ABSTRACT FROM AUTHOR]

Copyright of Iranian Journal of Veterinary Medicine is the property of University of Tehran and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

BACKGROUND: Staphylococcus aureus is one of the most important pathogenic microorganisms in meat products, especially those that are repeatedly handled by hand in the production process. Beta-lactam drugs, especially new generations of Cephalosporins, are used for treatment of most infections that are caused by Staphylococcus aureus. But the production of beta-lactamase enzyme by some strains has led to the failure for treating the infections that are associated with this organism. OBJECTIVES: The present study was conducted to evaluate the prevalence and comparison of the antimicrobial effect of methanolic extracts of red pepper and red onion on Staphylococcus aureus with beta-lactamase gene that was isolated from minced meat in Semnan city. METHODS: For this reason, sampling from 30 distribution and supply centers of packaged meat in Semnan city was performed in hygienic conditions and all of the samples were tested for presence of Staphylococcus aureus with betalactamase gene by biochemical methods and molecular confirmation by PCR assay. Also, the antibacterial effect of red pepper and red onion extracts on these isolates was evaluated by minimum inhibitory concentration (MIC), minimum bacterial concentration (MBC), well distribution and bacterial growth curve tests. RESULTS: The results showed that 16.6 percent of samples were contaminated with Staphylococcus aureus with beta-lactamase gene. Red pepper and red onion extracts had good antibacterial effects on these isolates and in all the tests, the red pepper extract was more effective than the red onion extract. CONCLUSIONS: By proving stronger antimicrobial effect of red pepper, it is recommended to use pepper in sufficient amounts along with onion in foods that are made from minced meat like all kinds of Kebab. [ABSTRACT FROM AUTHOR]

Copyright of Kafkas Universitesi Veteriner Fakultesi Dergisi is the property of University of Kafkas, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective(s): Escherichia coli strains are common pathogens that can cause urinary tract infections (UTI). This study aimed to assess E. coli phylogroups and virulence types in male and female UTI patients. Materials and Methods: In the present study, 160 uropathogenic E. coli (UPEC) isolates (from both sexes) were assigned to phylogroups/types and some extraintestinal virulence factors were detected within them by multiplex-PCR. Results: The isolates from women and men were predominantly distributed within phylogroup B2 and D, respectively. The presence of D2 phylotype was higher in men isolates than women, significantly (P=0.045). In male isolates papEF and sfa/focDE are more prevalent in B2 group than D, significantly (P=0.048; P=0.035). The prevalence of hly in B2 group is significantly higher than D (P=0.034) in female isolates. Conclusion: This study highlighted different features of E. coli genotypes from phylogenetic and virulence point of view implicated in UTI's in both human genders. [ABSTRACT FROM AUTHOR]

Aflatoxin B-1 (AFB1) is one of the major mycotoxins causing food contamination. Previous studies have shown that AFB1 can induce carcinogenicity and toxic effects in the isolated perfused rat liver and these effects are associated with its metabolites and peroxidation activity. Here we surveyed whether these pathogenic effects of AFB1 are associated with TNF-α as an inflammatory cytokine in general liver damages. In this study, we used twenty male Wistar rats (250-300 g). Rats were divided into four groups. Control group was pre-treated with LPS and then perfused with KHBB. The second group was pretreated with PTX and LPS and then perfused with KHB. The third group was pre-treated with LPS and then perfused with AFB-1 and KHB. The last group was pretreated with LPS and PTX and then perfused with AFB1 and KHB. Results revealed that aflatoxin B1 significantly increased the enzyme activity of aminotransferase and levels of lipid peroxidation. Also, the levels of Glutathione decreased in the aflatoxin group significantly. TNF-α released in perfusate and increased in aflatoxin B1 group significantly and decreased in AFB-1+PTX. Exposure to Aflatoxin B1 may induce reactive oxygen species, so these species may induce overproduction of proinflammatory cytokines such as TNF-α and may cause more damage to hepatic cells. [ABSTRACT FROM AUTHOR]

Background: This study was conducted to detect the prevalence of EHEC virulence genes and antimicrobial resistance profile of Escherichia coli strains belonging to B2 phylogroup implicated in Urinary tract infections in Semnan, Iran. Methods: From 240 urine samples 160 E. coli strains were isolated, biochemically. Then, E. coli isolates were examined by Multiplex-PCR for phylogenetic typing and detection of virulence genes (hly, stx1, stx2, eae) associated with Enterohemorrhagic E. coli. Finally, Antimicrobial resistance of E. coli isolates were characterized using Disk Diffusion method. Results: From 160 E. coli isolates, 75 strains (47%) were assigned to B2 phylogenetic group and prevalence of virulence genes were as follow: hly (21.3%), stx1 (16%), stx2 (10.6%) and eae (6.7%), subsequently. Phenotypic antimicrobial resistance of B2 isolates showed that all isolates were sensitive to Meropenem and Furazolidone and then highest frequency of resistance was observed to Streptomycin, Oxytetracycline, Neomycin, Nalidixic acid and Ampicillin (98.7% to 49.3%). Also low resistance prevalence was observed in case of Ceftizoxime, Lincospectin, Imipenem, Chloramphenicol and flurefenicole (16% to 1.3%). Conclusion: The data suggest a high prevalence of antibiotic resistance in UPEC strains belonging to B2 phylogroup even for the antimicrobials using in pet and farm animals and their potential to cause EHEC specific clinical symptoms which may represent a serious health risk since these strains can be transmitted to GI tract and act as a reservoir for other uropathogenic E. coli and commensal strains. [ABSTRACT FROM AUTHOR]

Background: Activation of inflammatory cells can cause more chemicals induced-hepatotoxicity. Aflatoxin B1 (AFB1) is a fungal toxin that induces acute hepatotoxicity in humans and animals. This study was conducted to examine the effect of co-exposure LPS and various aflatoxin B1 doses on the damage hepatic parameters in isolated perfused rat liver. Methods: Thirty-two male wistar rats (250-300g) were divided to eight groups including Control and LPS; three groups with various doses of AFB1 (0.01, 0.1 and 1 ppm) and three groups with various doses of AFB1 and Lipopolysaccharide (LPS) (300 ppm). Activity of Aspartate transaminase (AST) and Alanine transaminase (ALT) were determined in perfusate. Thiobarbituric acid reactive substances (TBARS) and Glutathione (GSH) concentrations were measured in homogenate liver. Results: At two groups of AFB 1 (with LPS and without LPS) at AFB1 concentrations of 0.1 and 1 ppm, elevation of AST and ALT enzymes activity were indicated. Values of GSH in both of groups (AFB 1 with LPS and without LPS) had reduced at concentration of 1 ppm. TBARS concentrations were enhanced in AFB1 concentration of 1 ppm in both of groups (with LPS and without LPS), however in comparison between groups (with LPS and without LPS) in similar concentrations significant different did not observe (P<0.05). Conclusion: Non-injurious dose of LPS did not enhance liver susceptibility to various doses of AFB1 in perfused rat liver. This may be in part of due to extrahepatic factors, which contribute, in more liver damage. [ABSTRACT FROM AUTHOR]

ABSTRACT Seventy-two 1-day-old broilers were allocated into four groups (basal diet as control, basal diet supplemented with 100, 200, and 400 mg/kg Zataria multiflora essential oil) and fed for 6 weeks. In the term, chicks were slaughtered and their breast fillets were stored at 4C for 12 days. Lipid peroxidation was assessed on the basis of thiobarbituric acid reactive substances formed, whereas microbial growth was assessed on the basis of total viable counts, lactic acid bacteria, Enterobacteriaceae and psychrotrophic bacteria. Results indicated that incorporation of Z. multiflora essential oil in broiler diets caused the delay of lipid peroxidation in raw breast meat (from 296.08 to 472.65 µg/kg malondialdehyde [MDA] equivalents) in comparison with 854.71 µg/kg MDA equivalents of the control sample at the last day of the experiment ( P < 0.05). Moreover, breast samples containing higher concentration of Z. multiflora essential oil showed lower bacterial count than the control and 100 ppm essential oil supplemented samples ( P < 0.05) during the experiment. PRACTICAL APPLICATIONS This paper evaluates the effect of dietary Zataria multiflora Boiss. essential oil supplementation on the microbial growth and lipid peroxidation of broiler breast fillets during refrigerated storage. It seems that Z. multiflora essential oil can delay the peroxidation and microbial spoilage of chicken breast fillets. Therefore, Z. multiflora essential oil might be used in novel applications as a nutritional supplement or a functional food component. [ABSTRACT FROM AUTHOR]

Aflatoxin B-1 (AFB1) is one of the major mycotoxins causing food contamination. Previous studies have shown that AFB1 can induce carcinogenicity and toxic effects in the isolated perfused rat liver and these effects are associated with its metabolites and peroxidation activity. Here we surveyed whether these pathogenic effects of AFB1 are associated with TNF-α as an inflammatory cytokine in general liver damages. In this study, we used twenty male Wistar rats (250-300 g). Rats were divided into four groups. Control group was pre-treated with LPS and then perfused with KHBB. The second group was pretreated with PTX and LPS and then perfused with KHB. The third group was pre-treated with LPS and then perfused with AFB-1 and KHB. The last group was pretreated with LPS and PTX and then perfused with AFB1 and KHB. Results revealed that aflatoxin B1 significantly increased the enzyme activity of aminotransferase and levels of lipid peroxidation. Also, the levels of Glutathione decreased in the aflatoxin group significantly. TNF-α released in perfusate and increased in aflatoxin B1 group significantly and decreased in AFB-1+PTX. Exposure to Aflatoxin B1 may induce reactive oxygen species, so these species may induce overproduction of proinflammatory cytokines such as TNF-α and may cause more damage to hepatic cells.