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.

Introduction: Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis, respectively the causative agents of bovine tuberculosis (bTB) and bovine paratuberculosis (PTB), share a high number of antigenic proteins. This characteristics makes the differential diagnosis of the diseases difficult. The interferon gamma (IFN-γ), C-X-C motif chemokine ligand 10 (CXCL10), matrix metallopeptidase 9 (MMP9), interleukin 22 (IL-22) and thrombospondin 1 (THBS1) bovine genes have already been shown to be accurate transcriptional biomarkers of bTB. In order to improve the diagnosis of bTB and PTB, in the present study we evaluated the risk of false positivity of these bTB biomarkers in cattle with PTB. Material and methods: The transcription of these genes was studied in 13 PTB-infected cattle, using Mycobacterium avium subsp. paratuberculosis (MAP)-stimulated peripheral blood mononuclear cells (PBMC). Results: Overall, the levels of IFN-γ, CXCL10, MMP9 and IL-22 transcripts in MAP-stimulated PBMC failed to differentiate animals with PTB from healthy animals. However, as bTB-afflicted cattle do, the MAP-infected group also displayed a lower level of THBS1 transcription than the non-infected animals. Conclusion: The results of this study add new specificity attributes to the levels of transcription of IFN-γ, CXCL10, MMP9 and IL-22 as biomarkers for bTB. | Instituto de Biotecnología | Fil: Klepp, Laura Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina | Fil: Klepp, Laura Ines. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina | Fil: Colombatti Olivieri, Maria Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina | Fil: Colombatti Olivieri, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina | Fil: Moyano, Roberto Damian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina | Fil: Moyano, Roberto Damian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina | Fil: Romano, Maria Isabel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina | Fil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina | Fil: Malovrh, Tadej. University of Ljubljana. Veterinary Faculty. Institute for Microbiology and Parasitology; Eslovenia | Fil: Ocepek, Matjaž. University of Ljubljana. Veterinary Faculty. Institute for Microbiology and Parasitology; Eslovenia | Fil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina | Fil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina | Fil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular (IABIMO); Argentina | Fil: Bigi, Fabiana. Consejo Nacional de investigaciones Científicas y Tecnológicas; Argentina

peer reviewed | [en] OBJECTIVE: Treatment of orofacial tumors in dogs is associated with high morbidity and reliable prognostic factors are lacking. Dynamic contrast-enhanced computed tomography (DCECT) can be used to assess tumor perfusion. The objectives of this study were to describe the perfusion parameters of different types of orofacial tumors and to describe the changes in perfusion parameters during radiotherapy (RT) in a subset of them. ANIMALS: 11 dogs with orofacial tumors prospectively recruited. CLINICAL PRESENTATION AND PROCEDURES: All dogs had baseline DCECT to assess blood volume (BV), blood flow (BF), and transit time (TT). Five dogs had repeat DCECT during megavoltage RT. RESULTS: 5 squamous cell carcinomas, 3 sarcomas, 1 melanoma, 1 histiocytic sarcoma, and 1 acanthomatous ameloblastoma were included. Blood volume and BF were higher in squamous cell carcinomas than in sarcomas, although no statistical analysis was performed. At repeat DCECT, 4 dogs showed a reduction in the size of their tumor during RT. Among these dogs, 3 showed an increase in BV and BF and 1 a decrease in these parameters between the baseline and the follow-up DCECT. The only dog whose tumor increased in size between the first and the second DCECT showed a decrease in BV and BF. CLINICAL RELEVANCE: Perfusion parameters derived from DCECT were described in a series of dogs with various types of orofacial tumors. The results suggest that epithelial tumors could have higher BV and BF than mesenchymal tumors, although larger sample sizes are needed to support these preliminary findings.