BACKGROUND Honey bee venom contains complex compounds such as polypeptides, enzymes, and amines. One of the important components of bee venom is the phospholipase A2 enzyme, which is considered an important honey bee venom allergen and is also used to treat some diseases. This enzyme is found in other insects, arachnids, snakes, and mammalian cells, and its function is the hydrolysis of the second ester bond of glycerophospholipids and the release of fatty acids and lysophospholipids. Although transient transfection can produce recombinant proteins, stable cells are more suitable for high-scale production with economic efficiency.OBJECTIVES: The present study created a stable cell line to produce recombinant phospholipase A2 from honey bee (Apis mellifera) venom.METHODS: Plasmid cloning DNA vector containing phospholipase A2 gene was prepared by Macrogen Company. The recombinant plasmid was transferred to Chinese hamster ovary cells by heat shock method, and gene expression was carried out in a HamsF12 culture medium containing neomycin antibiotic. After increasing polyclonal strains containing plasmid, monoclonal clones were selected by limiting dilution. Then, monoclonal clones were propagated, the soup of the selected cells was collected and concentrated, and the protein expression was checked by sodium dodecyl-sulfate polyacrylamide gel electrophoresis test.RESULTS: The results of electrophoresis, which was performed to confirm the expression of the phospholipase A2 gene in the cell soup, showed a band with a molecular weight of 20 kilodaltons, which confirms the creation of a stable cell line for the production of recombinant phospholipase A2 honey bee venom.CONCLUSIONS: After the transient transfection of the plasmid containing this gene, several cells undergo recombination due to having repair mechanisms and putting the desired gene along with the antibiotic resistance gene in their genome. These cells can be selected and propagated by adding antibiotics to the culture medium.

BACKGROUND: Toxocara canis is a zoonotic disease that commonly infects canids. Mammals and birds are sometimes infected with this disease as paratenic hosts. It can also cause accidental infection in humans. The increase in the number of stray dogs, the expansion of urban gardens, and the proximity of dogs to humans increase the risk of human infection with Toxocara canis.OBJECTIVES: This study aims to determine the prevalence of Toxocara canis infection in dogs and foxes in Zanjan province, Iran.METHODS: A total of 484 fecal samples of stray dogs (n=355), rescue dogs (n=49), guard dogs (n=50), and foxes (n=30) in Zanjan were randomly collected from June 2021 to February 2022. The microscopic examination was done following formalin-ethyl acetate sedimentation procedures. Finally, the PCR method was used to confirm the presence of Toxocara canis in positive samples.RESULTS: Microscopic study revealed that, out of 484 samples, 21 (4.3%) were positive for Toxocara/ Toxascaris eggs. Between these positive samples of dogs and foxes, only 6 samples from dog feces were confirmed as a Toxocara canis infection by the PCR method.CONCLUSIONS: There is an increase in the prevalence of Toxocara canis infection in stray dogs in Zanjan, Iran. Given the presence of dogs in parks and residential areas, there is a risk of human infection with Toxocara canis, emphasizing the importance of adhering to treatment and prevention protocols in dealing with stray dogs.

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: Excretory urography is a method of imaging the kidneys, ureters, and urinary bladder which uses contrast medium containing the iodine compounds.OBJECTIVES: This study aimed to evaluate the anatomical structures of urinary tract in the nephrogram, pilogram, and cystogram phases, and determine the exact standard for the size of kidneys, ureters and urinary bladder in guinea pigs to be used to interpret the results, and clinical decisions.METHODS: This study was carried out on 10 guinea pigs with a mean age of 12±1.33 months and average weight of 1.12±0.18 kg. Before to the administration of contrast medium, each guinea pig was fast and Dimethicone 20 mg/kg was given orally. At the time of administration of contrast agent, each animal was sedated by using Ketamine 30 mg/kg and diazepam 5 mg/kg cocktail, and then 1500 mgI/kg of meglumine compound 60 % was injected subcutaneously over the shoulder area. Ventrodorsales and lateral abdominal X-rays were taken, thereafter every 5 minutes up to 60 minutes to complete the pylogram phase. In lateral radiographs of each guinea pig, the length of the body of the second lumbar vertebra was measured to be used as an indicator in determining the standard size of the kidneys.RESULTS: Based on the results of this study, the average length, width, and thickness of the right kidney compared to the length of the second lumbar vertebra were 2.19, 1.64, and 1.33 cm, and in the left kidney of 2.09, 1.53, and 1.41 cm and this average in right and left ureter was 6.41 and 6.22 cm, respectively.CONCLUSIONS: The exact standards can be used in the interpretation of results, and clinical decisions to determine the normal and abnormal size of kidneys, ureters and bladder in the guinea pigs.

BACKGROUND: The protozoan Haemoproteus belongs to the Phylum Apicomplexa, Class Sporozoa, and Order Haemosporina. Avian haemosporidian are protozoan parasites that use birds as hosts around the world. Many species of wild and domestic doves are natural hosts of different species of Haemoproteus. Blood-sucking arthropods are the main vectors of these blood parasites.OBJECTIVES: The aim of this study was the microscopic and molecular investigation of the protozoan Haemoproteus columbae in the blood of infected pigeons in Torkaman County, Iran.METHODS: Blood samples and tubes containing ethylenediaminetetraacetic acid (EDTA) anticoagulant were collected from 96 domestic pigeons randomly from 14 pigeon lofts and different parts of Torkaman County.Pigeons were also inspected for infection with the host-vector Pseudolynchia canariensis. In the next step, blood smears were stained with Giemsa and examined microscopically. Also, blood tubes containing EDTA were tested by PCR method on the cytochrome b gene.RESULTS: Microscopic and molecular examination of peripheral blood showed that 62 (64.58 %) and 73 (76.04 %) of the investigated pigeons were contaminated, respectively. Of the 62 infected pigeons infected with the Haemoproteus, 28 pigeons (66.66 %) were male, and 34 (62.96 %) were female. Also, the infestation with Pseudolynchia canariensis was observed in 4 (28.57 %) pigeon lofts.CONCLUSIONS: The preliminary investigation shows the high rate of Haemoproteus infection in pigeons in Torkaman County. Further studies to determine the prevalence and accurate identification of the species infecting pigeons in this region require PCR testing and sequencing of infected blood samples.

BACKGROUND: Protection from reproductive damage is essential in chemotherapy medicines for cancer patients.OBJECTIVES: This study aims to examine the effect of curcumin on the structure of the ovary of mice after treatment with goserelin and cyclophosphamide.METHODS: One hundred and ten BALB/C mice with 3 regular consecutive periods of the estrous cycle were divided into 11 groups of 10 each. No medicine was used in the control group. The treatment groups were as follows: 1) cyclophosphamide, 2 to 5) cyclophosphamide with curcumin with a dose of 100, 200, 300, and 400 mg/kg, respectively, 6) goserelin, 7 to 10) goserelin together with curcumin with a dose 100, 200, 300, 400 mg/kg, respectively. The luteinizing hormone (LH) and follicle-stimulating hormone (FSH) of serums were evaluated using ELISA. Morphologic and morphometric of ovaries were assessed.RESULTS: The total number of follicles, primary, secondary, periantral, and antral follicles, in the goserelin and cyclophosphamide group, was significantly reduced compared with the control group (P<0.05). Cyclophosphamide and goserelin with different doses of curcumin showed a significant increase in the total number of follicles, primary, periantral, and antral follicles compared to the group treated with cyclophosphamide and goserelin alone (P<0.05). Curcumin (200, 300, and 400 mg/kg) and cyclophosphamide, compared to the cyclophosphamide group, significantly increased the quality of zona pellucida (P<0.05). Cyclophosphamide and goserelin caused a significant decrease in FSH and LH (P<0.05). Cyclophosphamide with different doses of curcumin showed a significant increase in LH compared to the group treated with cyclophosphamide alone (P<0.05). Goserelin with a 400 mg/kg curcumin dose significantly increased LH compared to goserelin alone (P<0.05). The amount of FSH in the cyclophosphamide groups with curcumin increased compared considerably to cyclophosphamide alone (P<0.05). The groups of goserelin with curcumin showed a significant increase in FSH compared to those of goserelin alone (P<0.05).CONCLUSIONS: Curcumin can protect the reproductive system of mice from the damage caused by the administration of cyclophosphamide and goserelin.

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: Gallibacterium anatis is a recently defined genus, which is a member of the Pasteurellaceae family. This advantageous pathogen is frequently found as part of the normal microflora of the upper respiratory tract and lower genital tract of the healthy poultry. Provided with appropriate conditions, it leads to various diseases, such as salpingitis, peritonitis, and loss of egg production with mortality in layer flocks. According to previous studies, multiple antibiotic resistance has been observed among G. anatis isolates, which can impose high costs on layer flocks. Due to the lack of the pathognomonic symptoms in the conflicts caused by this bacterium,  not enough comprehensive research has been conducted to date on the condition of this disease in Iran.OBJECTIVES: This study aimed to investigate the infection rates of this bacterium via PCR.METHODS: 295 tracheal swabs were collected from 10-layer flocks. Subsequently, the suspected colonies were isolated and identified with morphological features, differential cultivation, and PCR.RESULTS: 43.72 % of the samples were positive.CONCLUSIONS: The results of this study indicated that laying farms in Iran were infected with Gallibacterium anatis; thus, certain measures should be taken to control the factors reducing the production of layer flocks.

BACKGROUND: The widespread use of electromagnetic waves in communication has led to the consideration of their biological effects. According to valid scientific findings, these waves can entail changes in the cell function, there by causing or accelerating some diseases.OBJECTIVES: Since the amount of liver enzymes in the blood can be a sign of liver damage, this study was performed to evaluate these enzymes.METHODS: This experiment was done on 120 six-month-old pigeons. After adaptation to laboratory settings, they were divided into six random groups of distance from BTS and daily exposure time. G1: 50 cm/30 minutes, G2:100 cm/30 minutes, G3: 150 cm/30 minutes, G4: 50 cm/60 minutes, G5:100 cm/60 minutes, and G6:150 cm/60 minutes.Each group was exposed to waves for one month daily and blood tests were measured to determine the amount of liver enzymes before and one month after exposure; the results were processed by SPSS software.RESULTS: The findings of this study revealed a statistically significant relationship between BTS waves and liver damage after the increase in the level of liver enzymes in all six groups. Therefore, it is safe to assume that electromagnetic waves, depending on the frequency, energy, duration and distance, can have destructive effects on body tissues, including the liver. The mean changes of ALP and ALT were the highest and the lowest, respectively. Also, the most changes in liver enzymes occurred in the first and fourth groups, which had the shortest distance to the wave source.CONCLUSIONS: Increasing the exposure time and reducing the distance to the wave source changed the amount of liver enzymes. This study showed that "distance to the source of wave propagation" was more important than "exposure time".

BACKGROUND: Corynebacterium pseudotuberculosis and Trueperella pyogenes are two important pyogenic bacteria that cause many annual economic losses worldwide. Currently, antibiotic resistance of these bacteria is on the rise. Early detection of infection with these bacteria is important for controlling the infections caused by these two bacteria.OBJECTIVES: This study aimed to investigate the contamination of cattle cutaneous abscesses with  Corynebacterium pseudotuberculosis and Trueperella pyogenes in five large cattle farms around Tehran and propose an accurate method for a rapid detection of these two bacteria.METHODS: Out of 60 cows involved in cutaneous abscesses in the summer of 2018, sterile sampling was performed to diagnose the bacterial agent that caused the abscess. Bacteriological examination of the samples was performed using standard biochemical reactions and polymerase chain reaction using specific primers.RESULTS: Of the 60 samples studied, 25 % (15.60) were isolated as Corynebacterium pseudotuberculosis and 20 % (12.60) as Trueperella pyogenes. In 55 % of the samples (33.60), both bacteria were present simultaneously. All the samples detected using biochemical reactions were confirmed by the polymerase chain reaction.CONCLUSIONS: Corynebacterium pseudotuberculosis and Trueperella pyogenes are the main causes of cutaneous abscess in cattle farms around Tehran. Because the accurate diagnosis of the cause of abscesses is very important for effective treatment, polymerase chain reaction, based on 16S-23S rDNA and 16S rRNA, can be used to rapidly and accurately detect these bacteria in the early stages of the infection.