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: Infectious diseases and microbial antibiotic resistance are the major problems of fish farming industry annually causing remarkable losses. Apart from the economic losses caused by these infections, some of these agents are zoonotic and may be transmitted to humans.OBJECTIVES: This study was aimed to identify the common causative agents of infections in rainbow trout farms and to determine their antibiotic resistance toward some common antibiotics.METHODS: Sampling was performed during a nine-month period between March and December 2021 by visiting and inspecting rainbow trout farms and the affected fish with disease symptoms were obtained from the farmed fish in Mazandaran, Lorestan, Chaharmahal and Bakhtiari and Zanjan provinces. Bacterial culture was undertaken from anterior kidney or spleen organs and the isolated bacterial strains were identified by phenotyping, biochemical and molecular assays. Antibiotic resistance pattern was evaluated by disk diffusion method (DDM) and minimum inhibition concentration against erythromycin, oxytetracycline, florfenicol, enrofloxacin and nitrofurantoin.RESULTS: Seventy-four bacterial isolates of Gram-positive cocci or Gram-negative coccobacilli were isolated. In phenotyping, biochemical and molecular (PCR) assays Lactococcus garvieae (12 isolates, 16.2 %), Aeromonas hydrophila (9 isolates, 12.2 %), Streptococcus iniae (17 isolates, 23 %), Streptococcus agalactiae (20 isolates, 27 %), and Yersinia ruckeri (16 isolates, 21.7 %) were identified. The majority of these isolates were obtained from the fish farms in Mazandaran province. Erythromycin and oxytetracycline with 87.8 % resistance were antibiotics with the highest resistance, while enrofloxacin with 24.3 % resistance revealed the lowest level of resistance. Antibiotic resistance rates for florfenicol and nitrofurantoin were also 43.2 % and 44.4 %, respectively. The highest antibiotic resistance was detected in the bacterial isolates of Lactococcus garvieae, Aeromonas hydrophila, Streptococcus iniae, Streptococcus agalactiae and Yersinia ruckeri, respectively.CONCLUSIONS: This study shows that the spread of streptococcosis, lactococcosis, yersiniasis and Aeromonas septicemia and their frequent treatments has led to an increase in antibiotic resistance, especially against commonly used drugs such as erythromycin and oxytetracycline.

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: Clostridium perfringens (C. perfringens) is an anaerobic Gram-positive bacillus with spores, whose biotype A is responsible for a variety of diseases, including intestinal inflammation, bloody diarrhea, and gas gangrene, and hemorrhagic bowel syndrome. Genetic variety can explain the bacteria’s phenotypic diversity, geographic distribution, host specificity, pathogenicity, antibiotic resistance, and virulence. A molecular method using the pattern of DNA bands classifies bacteria based on the size of fragments produced by enzymatic digestion of the genome.OBJECTIVES: This study aims to standardize the polymerase chain reaction (PCR)- restriction fragment length polymorphism (RFLP) method in identifying the genetic diversity of C. perfringens biotype A isolates.METHODS: The genomic DNA of the investigated strains was extracted, and the complete sequence of the alpha toxin gene locus was synthesized using specific primers designed by PCR technique. Enzymatic cleavage of the synthesized amplicons was performed with the Mse l restriction enzyme, and the resulting fragments were separated by electrophoresis and analyzed by ImageJ and NTSYSPC software.RESULTS: The findings showed that the alpha toxin gene locus sequence may change and is not conserved. In this research, 4 different patterns were identified based on enzymatic cleavage. Mutations in this locus can lead to diversity in C. perfringens biotype A and the creation of new strains.CONCLUSIONS: The results of this research showed that the alpha toxin gene locus could be considered a DNA molecular marker in C. perfringens, and the PCR-RFLP technique can be used as a tool for typing this bacterium and estimating the phylogenetic relationships through comparative studies of nucleotide sequences.

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: Salmonellosis is a dangerous disease that can threaten the health of humans and animals. This disease can lead to economic losses annually; therefore, many studies have been conducted to prevent this disease.OBJECTIVES: The current study aims to design a recombinant chimera protein HBHA-Omp28 as a vaccine against Salmonella typhimurium.METHODS: The nucleotide and amino acid sequences of Omp28 and HBHA proteins were first extracted from the NCBI database. Then, the recombinant chimera of HBHA-Omp28 was bioinformatically assembled using a rigid linker. Epitope prediction of T and B cells, antigenicity, allergenicity, and physicochemical features assessments of HBHA-Omp28 were done using Immune Epitope Database (IEDB), ABCpred, VaxiJen, AllerTOP and ProtParam online servers, respectively. To assess the secondary and tertiary structures, the Self-Optimized Prediction Method with Alignment (SOPMA) and the Iterative Threading ASSEmbly Refinement (I-TASSER) server were used, respectively. Molecular docking between recombinant chimera and TLR4/MD2 receptor was assessed by ClusPro server. Finally, after codon optimization of nucleotide sequence of recombinant chimera to express in Escherichia Coli k-12 strain, the cloning of recombinant chimera in pET21-a (+) vector was examined.RESULTS: The designed recombinant chimera was classified as an antigenic and non-allergenic protein with molecular weight of 34.19 kDa. According to the results of molecular docking study, the HBHA-Omp28 protein was able to bind to TLR4/MD2 receptor using 9 hydrogen bonds. The results of cloning study demonstrated that HBHA-Omp28 successfully cloned into pET21-a (+).CONCLUSIONS: The designed recombinant chimera can be an appropriate vaccine against salmonella bacteria.

BACKGROUND Agricultural pesticides can cause environmental pollution and damage to aquatic organisms. Bensulfuron-methyl is a widely used herbicide in agricultural fields, especially rice fields. Despite the solubility of Bensulfuron-methyl in water and its entry into aquatic environments, limited research has been conducted on the toxicity of this herbicide in aquatic organisms.OBJECTIVES: This study aims to investigate the effects of chronic toxicity of Bensulfuron-methyl in common carp (Cyprinus carpio).METHODS: The fish were divided into four groups. Group 1 was considered as a control, and groups 2, 3, and 4 were exposed to 10 %, 20 %, and 30 % of the 96 h lethal concentration 50 of Bensulfuron-methyl equal to 0, 0.162, 0.324 and 0.486 g/L.  After 21 days, blood samples, serum levels, and liver tissue of fishes were analyzed.RESULTS: The number of white blood cells increased in groups 2 and 3 (received 0.162 and 0.324 g/L Bensulfuron-methyl) compared to group 1, while a significant decrease was observed in group 4 (received 0.486 g/L Bensulfuron-methyl) compared to other groups. The number of red blood cells, the amount of hemoglobin, and the percentage of hematocrit in groups 3 and 4 showed a significant decrease compared to other groups, and the values of mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were not significantly different in any groups. The amount of total serum protein in groups 3 and 4 decreased significantly compared to the control group. Serum glucose showed a significant increase in groups 3 and 4 compared to other groups. The values for aspartate aminotransferase, alanine transaminase, and alkaline phosphatase enzymes showed an increasing trend with the increase of Bensulfuron-methyl concentration. The most liver tissue damage was observed in group 4, which included hyperemia, hepatocyte vacuolar degeneration, edematous cell infiltration, bile duct hyperplasia, and hepatic necrosis.CONCLUSIONS: The increase in the concentration of Bensulfuron-methyl can cause liver tissue damage and changes in hematological and serum biochemical markers in common carp.

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.

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.