BACKGROUND: The effect of nanoherbal substances in the treatment of infectious diseases before mating or during pregnancy must be evaluated. Saponin has adverse effects on a pregnant mother and her fetal through complex mechanisms.OBJECTIVES: This study aims to investigate the toxic effects of saponin encapsulated by ferritin nanoparticles (Nanosaponin) on fetal lung development in female mice with Streptococcus pneumonia.METHODS: Extraction of crude saponin from licorice roots was done by the powder heating and using 20 % ethanol and diethyl ether. Then, n-butanol and 5 % sodium chloride solution were added to the aqueous layer. The mice were divided into five groups of 10 including control, pneumonia, pneumonia treated with ferritin, pneumonia treated with saponin, and pneumonia treated with nanosaponin. Then, two groups of untreated pregnant pneumonia and pregnant pneumonia treated with nanosaponin were separated. In this study, tumor necrosis factor alpha (IFN-γ), heparin-binding epidermal growth factor-like growth factor (HB-EGF), Methyl-CpG binding domain 2 (MBD-2), and kruppel like factor 2 (KLF-2) genes were evaluated in the maternal lungs, and TNF-α level was evaluated using ELISA method in the fetal lungs. Maternal tissue, uterus tissue, and fetal lung were examined by hematoxylin and eosin staining, and maternal body tissue was examined by trichromason staining and Oxidative stress parameters were investigated.RESULTS: Nano saponin decreased the expression of IFN-γ (P<0.05) (P<0.01), MBD-2 (P<0.05) and HB-EGF  genes, KLF-2 protein level and TNF-α levels in fetal lung.The thickness of uterine myometrium and blood flow of endometrium increased the number of live embryos and the rate of successful pregnancy. In addition, Nano saponin effectively improved the oxidative stress response in the fetus without any harmful effect on the mother's liver tissue.CONCLUSIONS: The nanosaponin has no toxic effects on the sudy organs of the mother and fetus. It has therapeutic effects on the fetal lung development process after the infection of mother with Streptococcus pneumonia.

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: Body condition and its relationship with inflammatory indicators in the transition period of ewes can be used as a key to prevent the occurrence of metabolic complications in this period.OBJECTIVES: This study aims to determine the levels of inflammatory markers and their relationship with body condition and energy status in the transitional period in Makuei ewes.METHODS: This study was performed on 45 female peri-parturient Makuie ewes aged 3-5 years with 2-4 breeding lambs. Blood samples from the jugular vein were prepared in three periods, 21 days before delivery, baseline (time zero), and 21 days after delivery.RESULTS: The mean glucose and cholesterol concentrations were not significantly different between the groups with low, moderate, and high body conditions. Non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) concentrations were significantly higher in groups with lower and higher body condition scores (BCS) than in the normal group. There was a significant positive correlation between energy-related indices (NEFA, BHB) and the BCS of the pregnant and lactating ewes. The concentration of fibrinogen, sialic acid, and blood ceruloplasmin increased in the first three weeks and decreased after delivery. These indices significantly increased in relatively obese and lean groups than in the normal group during the study. The correlation of BHB and NEFA with sialic acid, ceruloplasmin, and fibrinogen was also reported in the study groups in the pre- and post-partum periods.CONCLUSIONS: Ewes with normal BCS (2.75-3.25) have a good energy status. Low levels of NEFA in ewes indicate that the mobility of fats is low, and the inflammation process is lower in the transition period of these animals. Furthermore, low BCS can be a predisposing factor for inflammation in ewes during the pre-partum period. This effect may be due to the increased metabolic requirements and compromised immune function associated with negative energy balance in the transition period of ewes.

BACKGROUND: Cadmium is a heavy metal harmful to animals and humans. Exposure to it causes inflammation, apoptosis, or necrosis in numerous tissues, including the adrenal.OBJECTIVES: The present research investigates the effect of cadmium toxicity on the expression of genes involved in inflammation and fibrosis. Inflammation increases the rate of parenchymal cell death, and fibrosis will only fill the place of dead cells without being able to perform the function of the primary parenchyma.METHODS: In this research, cadmium chloride with a concentration of 20 mg/kg was added to the diet of ten mice in two groups of five. On the 30th day of the study, the adrenal glands were quickly sent to the laboratory. The expression of NF-kB/MAPK, hematoxylin, eosin tissue staining, and immunohistochemistry (CD163) were performed.RESULTS: The inflammation mentioned in others’ research can also be associated with the activation of the nuclear factor kappa (NF-kB) pathway. NF-κB gene products initiate mitogen-activated protein kinase (MAPK) and p38 pathways. Previous studies indicate that MAPK induces necrosis or apoptosis in tissues. In histopathology, dense and possibly pyknosis nuclei are more common in the cadmium group. The higher expression of the CD163 molecule in the cadmium group reveals the beginning of the fibrosis process after chronic inflammation.CONCLUSIONS: This report provides more basic data to investigate the mechanism of adrenal damage in cadmium poisoning. Cadmium causes the death of cells by affecting the inflammatory pathways. Additionally, the stimulation of the fibrosis process causes greater irreparable damage to the damaged tissue of the adrenal gland.

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: Numerous studies have investigated the positive effect of chitosan and nano-chitosan loaded clinoptilolite on growth performance, digestive enzyme activity, and intestinal histomorphology in different fish species. However, there are no data evaluating the potential effect of the composites on the fish stomach.OBJECTIVES: In the current study, the effects of chitosan and nano-chitosan loaded clinoptilolite on histological features and pepsin activity in the rainbow trout stomach were considered.METHODS: Chitosan and nano-chitosan loaded clinoptilolite were synthesized, and then two hundred and forty fish (~27.75 g) fish were distributed in eight groups each in three replicates. Ten days after adaptation, the fish were fed with eight diets, including control diet (CTR), clinpotilolite (T1), chitosan composites (T2, T3, T4), and nano-chitosan composites (T5, T6, T7) for 70 days. Afterward, all fish in each tank were anesthetized in clove extract (50 μl/l), and tissue samples were obtained for pepsin activity (n= 5) and histological assay (n = 5).RESULTS: The groups administrated with nanochitosan composites showed the highest pepsin activity (P˂0.05). Additionally, histological examinations exhibited a higher epithelial height, increased mucosal density, and oxynticopeptic cells hypertrophy in fish fed composites compared to the CTR group (P˂0.05). Meanwhile, nanochitosan composite administration could cause higher reaction of secreted granules to periodic acid–Schiff (PAS) staining.CONCLUSIONS: The findings demonstrated the potential application of chitosan and nano-chitosan loaded clinoptilolite composites for improvements in the histomorphology and pepsin activity of the rainbow trout stomach, resulting in higher growth performance.

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: Milk and dairy products are important sources of food-borne pathogens. Non-pasteurized dairy products are popular due to home production, beliefs about their higher nutritional value, high accessibility, and taste.OBJECTIVES: This study aims to investigate the consumption pattern of local dairy products in women in Qom, Iran, in 2022, and determine the affecting factors.METHODS: In this cross-sectional study conducted in 2022, 319 women in Qom were selected using a stratified random sampling method. Their demographic information (age, educational level, employment status, and income) and consumption of local dairy products were surveyed. In addition, a questionnaire based on the theory of planned behavior (TPB) with 32 items and 4 subscales (attitude towards nutrition, subjective norms, behavioral intention, and nutritional behavior) was completed. The data was analyzed in SPSS software using ANOVA, and Chi-square test.RESULTS: Overall, the consumption rate of local milk was 82.3 %; yogurt, 85.1 %; cheese, 57.3%; cream, 53.7 %; butter, 42.3 %; and curd, 33.9 %. Regarding the daily consumption rate, the highest consumption rate was related to milk (13.9 %) and yogurt (11.8 %), and the lowest consumption was related to curd (3.1%) and cream (5.1 %). The type of dairy consumed was significantly related to behavioral intention and nutritional attitude (P<0.05). There was a significant difference in the type of consumed dairy in terms of the husband's occupation (P=0.001), but there was no significant difference in terms of educational level, marital status, employment status, and relationship with the villagers (P>0.05).CONCLUSIONS: The prevalence of local dairy products consumption, especially milk and yogurt, is high in women living in Qom. Their behavioral intention to consume healthy dairy products is at good level, but they do not have proper nutritional attitude and nutritional behavior. Therefore, the risk of developing common zoonotic diseases, including brucellosis and crimean-Congo hemorrhagic fever is high in Qom.

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.