BACKGROUND: Poultry farming is one of the most productive and economic agricultural sectors. However, the bacterial contamination and the activity of darkling beetles (Tenebrionidae) as a potential reservoir of Salmonella in meat poultry farms can inflict direct and indirect damages.OBJECTIVES: The present study aimed to identify the darkling beetles and their accompanying Enterobacteriaceae contamination in Isfahan chicken farms.METHODS: Darkling beetles were collected and identified based on their morphological aspects from different parts of 16 poultry farms (4 from each geographical area) in Isfahan Province, Iran. Then, 80 samples of darkling beetles were cultured on selective-differential media culture of the Enterobacteriaceae family using the homogenization and enrichment method. The isolated bacteria were identified based on physiological and molecular characteristics. Also, specific antisera were used to determine serological groups.RESULTS: The results revealed that all collected darkling beetles’ samples belonged to the species Alphitobius diaperinus (Col., Tenebrionidae), and from 80 microbial culture samples from the beetles, isolated bacteria belonged into 4 genera: Escherichia sp. (20 isolates, 25 %), Klebsiella sp. (8 isolates, 10 %), Proteus sp. (22 isolates, 27.5 %), and Salmonella sp. (30 isolates, 37.5 %). Among them, the Salmonella genus accounted for the highest percentage of darkling beetles’ contamination. In the serological assay, the isolated Salmonella were classified into two serogroups, A (23 isolates, 76.67 %) and C (C2 and C3) (7 isolates, 23.33 %), which the A serogroup was the most frequent.CONCLUSIONS: In this study, the A. diaperinus species was isolated and identified for the first time from poultry farms, and this pest, with a high percentage of Salmonella infection, is introduced as one of the reservoir sources of bacterial contamination in the broiler farms.

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 subjected 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 subjected 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.

BACKGROUND: Lyme disease is caused by a gram-negative spirochete bacterium called “Borrelia burgdorferi” and can be transmitted by the bite of infected ticks to humans and animals. This disease has a global geographic distribution Dogs can be infected by the bite of the Ixodes ricinus tick.OBJECTIVES: This study aims to detect the prevalence of Borrelia burgdorferi in guard dogs in Isfahan, Iran.METHODS: In this study, blood samples were collected from 97 guard dogs with an average age of 3.5 years in Isfahan city and analyzed by the blood smear method and the nested polymerase chain reaction method (molecular study). The data were statistically analyzed in SPSS software (version 24) using Fisher's exact test and chi square.RESULTS: Twelve samples (12.37 %) were found molecularly positive, which were for 10 male dogs (10.30 %) and two female dogs (2.06 %). There was no evidence of the presence of bacteria in the blood smear and hematological changes were not found in complete blood count tests (CBC Test) performed on infected samples. There was a significant difference in the levels of infection between the age groups of 1 to 2 years (P=0.029) and between this age group (1 to 2 years) and the age group >3 years (P=0.032. (There was a significant difference in infection levels between dogs with different gender.CONCLUSIONS: The prevalence of Borrelia burgdorferi in dogs of Isfahan City is relatively high. Due to the ease of transmission of this pathogen between humans and animals, special attention should be paid for its control and timely detection.

BACKGROUND: Ischemia-reperfusion injury (IRI) can induce major changes in the function of different organs, including the liver. Studies have indicated that ischemic post-conditioning (HIPO) can protect the tissues against ischemia-reperfusion injury.OBJECTIVES: To investigate the antioxidant and anti-inflammatory effects of ischemia post-conditioning against the IRI of the rat liver through four 30-second cycles of alternating ischemia and reperfusion, before 24-hour persistent reperfusion.METHODS: Fifteen rats were randomly divided into 3 groups 1) operation control group, 2) ischemia-reperfusion (IR) group whose liver was exposed to 60-miute ischemia of by 24-hour reperfusion and 3) ischemic post-conditioning (IR+IPO) group underwent the same procedure as the second group except that before persistent reperfusion, the rats were subject to post-conditioning by four 30-second cycles of alternating ischemia and reperfusion. The changes induced by IRI and the antioxidant and anti-inflammatory effects of ischemic post-conditioning were assessed by the serum level of IL-6 using the ELISA method, malondialdehyde (MDA), and total antioxidant capacity (TAC). RESULTS: Ischemic post-conditioning potentiated antioxidant effects and reduced the inflammation caused by the IR in the liver. The serum level of IL-6 reduced from 394.4±126.4 to 124.4±29.07 pg/ml (post-conditioning group), and the tissue MDA reduced from 431.4±76.53 to 207.2±25.77 nmol/g) compared to the IR group. The data revealed that the levels of the indices returned almost to the level of the operation control group (P<0.001). Additionally, the total antioxidant capacity of the liver significantly improved (P<0.01) from 11.58±1.87 (in the IR group) to 17.53±2.51 mmol/mg (in the IR+IPO group).CONCLUSIONS: The study demonstrates that the protective effect of ischemic post-conditioning against IR-induced injury may be mediated through decreasing inflammation and improving antioxidant activities.

BACKGROUND: Recently, a subpopulation of monocytes expressing Tie2 receptors has been identified, playing an important role in tumor angiogenesis. Selective depletion of Tie2-expressing monocytes in tumor-bearing mice can inhibit tumor angiogenesis. Some of these macrophages have been shown to be located near the tumor blood vessels, forming vessels in these areas paracrinely during the angiogenesis process.OBJECTIVES: This study aims to investigate the role of putative phosphorylation sites on Tie2 receptor in tumor- associated macrophages connected to human endothelial cells.METHODS: In this study, we used a series of Tie2 mutants. After transfection of tumor-associated macrophages  with these mutants, they were evaluated for physical connection using the surface plasmon resonance technique and by non-contact co-culture of these macrophages with endothelial cells.RESULTS: Mutation in the tyrosine residues 1106 and 1111 had an inhibitory effect on macrophage binding to endothelial cells, resulting in deterioration of the angiogenic activity of these cells.CONCLUSIONS: Tie2 receptor and its downstream molecular pathways such as AKT/PI3 have a role in the interaction of tumor-associated macrophages with human endothelial cells, directly (via physical binding) and indirectly (through secretion of factors affecting angiogenesis). This emphasize the importance of the molecular mechanisms of Tie2 receptor activation in the interactions of endothelial cells with tumor-associated macrophages and as an anti-angiogenic therapy for cancer.

BACKGROUND: Nowadays, the interest in functional food has dramatically increased. Herbal plants and functional foods have health-enhancing effects on consumers due to their medicinal, antioxidant, and nutritional properties. Probiotics are one of the most emerging and popular functional food products. OBJECTIVES: This study aims to assess the effect of irradiated and non-irradiated saffron petal extract on the viability of Lacticaseibacillus paracasei (M4PM99) in probiotic yogurt.METHODS The ethanolic extract of irradiated saffron petals with a 10 KGy dose of gamma ray at concentrations of 25, 50, and 75 mg/mL and non-irradiated extract at the same concentrations were used and their effect on the viability of Lacticaseibacilus paracasei and their antioxidant and physicochemical properties in set yogurt were studied. Probiotic survival, pH, acidity, content of total phenolic compounds, DPPH inhibition percentage, and sensory properties on days 0, 7, 14, and 21 were assessed.RESULTS: Both irradiated and non-irradiated saffron extracts significantly increased the viability of probiotic bacteria compared to the control sample (P<0.05). The addition of extracts was effective in increasing acidity and decreasing pH compared to the control (P<0.05). With the increase in the amount of extract, the percentage of DPPH inhibition and phenolic compounds significantly increased in the irradiated samples (P<0.05). The effect of storage time was also significant on these indicators, such that the antioxidant properties and phenolic compounds increased until the 14th day and then decreased (P<0.05). In the sensory evaluation, in terms of taste, odor, and color, the lowest score was related to the sample containing 0.75% extract. No significant difference was observed in other concentrations compared to the control sample.CONCLUSIONS: Saffron petal extract has a positive effect on the viability of probiotics during storage. Gamma irradiation has a significant effect on the amount of total phenolic compounds and the antioxidant activity of saffron petal extract. It can be used as a natural antioxidant in dairy products.

BACKGROUND: The H9N2 avian influenza is one of the most important viral diseases of poultry in Asian countries. Continuous mutations in the hemagglutinin (HA) protein as the main viral antigen make interpreting the HA-based serology test results difficult.OBJECTIVES: In this experimental study, the ability of polyclonal antibodies against the conserved motif of HA protein in detecting heterologous H9N2 isolates was evaluated.METHODS: Based on the bioinformatics data, a conserved motif of HA protein was selected and expressed in the pET-28a(+) vector. The recombinant peptide was mixed with an oil adjuvant and injected into specified pathogen free (SPF) chickens to produce the polyclonal antiserum. Similarly, antiserum from the inactivated virus was prepared. The efficiency of the prepared antisera was evaluated in the cross hemagglutination inhibition (HI) test using the homologous and heterologous H9N2 viruses.RESULTS: The average titer of HI antibody against homologous and heterologous viruses using the polyclonal antiserum was equal to five. This titer was estimated to be seven for the homologous virus and much lower for heterologous viruses with inactivated virus antiserum. The difference was statistically significant.CONCLUSIONS: According to the evaluations, this antiserum has a suitable efficiency in identifying different isolates of the H9N2 virus, and compared to the usual antiserum, it detects different antigens of a subtype more accurately. The resulting polyclonal antiserum does not contain non-specific inhibitors, which may cause problems interpreting serological findings. The results showed that polyclonal antiserum could identify H9N2 viruses isolated in different years by HI test; however, its validation requires testing with more viruses.

BACKGROUND: Temporomandibular joint (TMJ) is created by the connection between the mandibular condylar process and the mandibular fossa of the temporal bone in all mammalian species. This is a synovial joint and has a fibrocartilage disc. The joint disorders affect mastication, movements of TMJ, and mandibular functions. TMJ imaging can be evaluated by radiography; however, CT imaging is preferred, since the images have no superimposition and present more details of different structures and therefore show a more accurate evaluation of TMJ.OBJECTIVES: This study aimed to provide accurate statistics on TMJ disorders and to evaluate their prevalence based on CT-scan findings in dogs and cats.METHODS: In this study, 265 cases of cats and dogs (102 cats and 163 dogs) were evaluated. A total of 59 cases (38 cats and 21 dogs) had temporomandibular joint disorders. In this study, information on sex, breed, and age regarding TMJ disorders were described in detail.RESULTS: In the evaluated CT images, osteoarthritis, fracture, dislocation (luxation and subluxation), ankylosis, osteochondrosis, dysplasia, and osteomyelitis were observed.CONCLUSIONS: This study indicated that although the total number of dogs was larger than that of cats, the incidence of temporomandibular joint disorders was higher in cats than in dogs. The most common disorders in cats and dogs were fractures, followed by dislocations (luxation and subluxation), and then osteoarthritis exhibited the highest frequency. In the next rank of frequency in cats, ankylosis and then osteomyelitis were observed. Additionally, osteochondrosis and dysplasia and then ankylosis were observed in dogs in the next ranks.

BACKGROUND: 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.OBJECTIVES: 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.METHODS: 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.RESULTS: 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.CONCLUSIONS: 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.

BACKGROUND: Ovine pulmonary adenocarcinoma, also known as jaagsiekte, is a chronic, contagious, and transmissible lung cancer. It is prevalent in ovine spp while rarely occurring in caprine with long incubation period. The disease is mostly observed in older animals (over 2 years old).OBJECTIVES: The present study aims to determine the prevalence of the disease based on histopathological diagnosis and investigate its correlation with age and sex in the slaughtered local sheep in Garmsar (Semnan province).METHODS: Herein, the lungs of 9030 slaughtered sheep are morphopatthologically examined for the presence of ovine pulmonary adenocarcinoma.RESULTS: Based on the morphopathological examination, ovine pulmonary adenocarcinoma was observed in the lungs of 438  (4.87 %) out of 9030 sheep. That said, 250 indicated the classical form, as firm, white to grayish coalescing masses mostly in the cranio-ventral lobes; this form is associated with wet cut surface and frothy fluid in the airways. The remaining 188 sheep showed the atypical form, as small, clear demarcated nodules mostly in diaphragmatic lobes associated with dry cut surface and minor fluid in the airways. Almost similar histopathological changes were seen in the two forms. An acinar or papillary growth of neoplastic cells in the alveoli and polypoid proliferation of bronchiolar epithelium were observed in both forms. However, there were variable amounts of connective tissue, and infiltration of lymphocytes and plasma cells in the interstitial tissue of the affected alveoli and no metastatic lesion in the lymph nodes. The peribronchial and peribronchiolar lymphoid aggregates were consistent features in most of the cases studied.CONCLUSIONS: The findings of the present study, as the first report of ovine pulmonary adenocarcinoma in sheep from Garmsa county, revealed the higher prevalence of this disease compared to that reported in previous reports in the country. Moreover, according to the obtained results, atypical and classical forms represented different stages of a single disease spectrum.