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: In free stalls, factors related to the surface and dimensions of the stall affect how the cows rest and comfort. The brisket board and the neck rail are the most controversial parts of the free stall in Iran's dairy farms, that can affect the stability of the stall and its lifespan, while improper use of these structures has led to significant discomfort for cows, causing substantial issues including lameness and hock, knee and withers lesions.OBJECTIVES: This study aims to investigate Brisket boards and neck rails usage and measures in freestall barns and assess its possible impact on some comfort indices in dairy cows.METHODS: Nine dairy farms with over 100 milk cows and freestall barns were selected using the Dairy Farmers of Canada protocols by a convenience sampling method. Horizontal, vertical, and diagonal distances of the neck rail, the presence or absence of brisket boards, and the brisket board height from the bedding were measured. The locomotion score based on a five-point scale as well as hygiene, knees, hocks, and withers scores were recorded. The correlation was evaluated using the Spearman correlation test and Pearson’s correlation test.RESULTS: In 68.3 % of the freestall barns, the brisket boards were at the bedding level or were not used at all; however, the mean brisket board height (11.2±10.8) was not significantly different from the standard height value of 10 cm (P>0.05). The vertical distance of the neck rail (120.4±10.4 cm) was significantly different from the standard values. The median of withers and locomotion scores were consistent among all farms. At the farm level, the median knee, hygiene, and hock scores did not show a significant correlation with the mean of neck rail measures and brisket board height (P>0.05). Also, the median locomotion score did not show a significant correlation with the mean horizontal distance of the neck rail at the individual freestall barn level (P>0.05). However, a significant correlation between the mean of knee scores and vertical distance of the neck rail at the farm level, and between the mean of locomotion score and horizontal distance of the neck rail at the individual freestall barn level were reported.CONCLUSIONS: An increase in the mean vertical distance of the neck rail is associated with an increase in the median knee scores, while an increase in the mean horizontal distance in each barn was associated with an increase in the median locomotion score, indicating the potential impact of these measurements on cow comfort. However, further research using a larger sample size is needed.

BACKGROUND: Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis (CLA), a chronic and very common disease in sheep and goats, which can lead to severe economic losses in the livestock industry.OBJECTIVES: This study aims to investigate the prevalence of CLA in sheep in Kurdistan province of Iran using phenotypic and molecular methods, and assess the antibiotic resistance of isolated Corynebacterium pseudotuberculosis.METHODS: In this study, from September to March 2022, 270 samples of skin abscesses were collected from sheep in livestock farms of Kurdistan province. Immediately, using the cold chain system, the samples were transferred to the microbiology laboratory of the Faculty of Medicine at Kurdistan University of Medical Sciences. Identification of isolates was done using biochemical tests and polymerase chain reaction (PCR) method. The antibiotic resistance of the isolates was examined using the Kirby-Bauer disk diffusion method.RESULTS: Based on biochemical tests, out of 270 samples, 82 suspected to have Corynebacterium pseudotuberculosis. Out 82 samples, the presence of bacteria was confirmed in 76 samples by the PCR. The antibiotic sensitivity test showed that the isolates had high sensitivity to doxycycline and ceftriaxone and high resistance to streptomycin and kanamycin.CONCLUSIONS: The CLA has a high prevalence in sheep in Kurdistan province. According to high resistance rate of Corynebacterium pseudotuberculosis to streptomycin and kanamycin, it recommended to avoid treatment of CLA cases with these antibiotics.

BACKGROUND: Based on the historical and recently published documents, it has been demonstrated that Mirabilis jalapa as a herbal medicine may be used as remedies for various health problems included wound healing purposes.OBJECTIVES: The present study aimed to investigate the effect of ethanolic extract of Laleh abbasi green leaf on healing open wounds induced by skin puncture in the back of rats.METHODS: Collecting and drying Laleh abbasi leaves, leaf extracting through Soxhlet procedure, analyzing the extract via gc / ms method, and preparing eucerin-based extract ointment were done according to recommended routine procedures. Herein, we recruited 40 male rats that were randomly divided into five groups of eight, namely the control, phenytoin treatment, eucerin, 5% eucerin extract, and 7.5% eucerin extract ointment treatment groups. Skin puncture and application of ointments on the induced wounds were carried out. Subsequently, tissue samples were taken on days 3, 7, 10, and 14, followed by which histological slides were prepared and stained with H&E and Masson’s trichrome staining methods. In vitro mycological studies were conducted using opportunistic fungi, including Candida, Mucor, and Aspergillus species.RESULTS: Based on the macroscopic evaluations of the wound healing process and microscopic assessments of tissue samples stained with Harris H&E and Masson’s trichrome methods, the groups treated with eucerin-based ointments containing ethanolic extract of Laleh abbasi leaf had statistically significant positive wound healing responses compared to the other groups. However, the 7.5% ointment group showed statistically better responses compared to the 5% ointment group. The data obtained in the present preliminary experiment on rat model indicated that the extract could facilitate the wound healing process in terms of healing parameters, such as accelerating epithelium repair, creating a favorable inflammatory reaction, angiogenesis, fibroplasia, and collagen precipitation. The antifungal effects of ethanolic extract of Laleh abbasi leaves on Aspergillus fumigatos, fusarium, Candida albicans and Candida cruzei were demonstrated in vitro using saboro dextrose agar medium.CONCLUSIONS: According to the findings of this experimental study and the findings of other researchers, it can be concluded that ethanolic extract from Laleh abbasi leaves is of healing and antifungal properties.

BACKGROUND: Mycobacterium avium subspecies paratuberculosis is the cause of a common disease in dairy herds. Early diagnosis of paratuberculosis infection can improve Johne’s disease control programs.OBJECTIVES: This study aimed to compare the sensitivity, and specificity to methods; blood serum ELISA and stool Nested-PCR for the detection of Mycobacterium avium subspecies paratuberculosis infection in dairy cattle.METHODS: A commercial ELISA kit was used to perform the absorbed ELISA test, which was conducted after exposing serum samples to Mycobacterium phlei antigens to limit cross-reactions. Nested-PCR test was performed using nucleotide sequences related to specific MAP gene fragments, i.e. IS900.RESULTS: As a result of the ELISA antibodies kit, out of the total 2203 serum samples, 112 samples were positive (5.08 %) and 2091 samples were negative (94.92 %). The results of Nested-PCR tests of rectal feces showed that out of 59 cows with the positive results in serum ELISA, 47 (79.66 %) samples were positive and 12 (20.34 %) samples were negative. Moreover, out of 31 cattle with a negative result on the ELISA test, 15 (48.38%) and 16 cattle (51.62 %) had positive and negative results, respectively, on the nested PCR tests of the feces samples.CONCLUSIONS: Due to the low sensitivity of PCR compared to ELISA, the positive and negative predictive values, and the accuracy of ELISA test, as well as the high cost and time-consuming nature of PCR and the need for more and more complex facilities than ELISA, the authors concluded that ELISA is a more suitable method for screening and epidemiological studies than PCR.

BACKGROUND: Horses are economically and emotionally valuable animals in various activities, especially sports. Thus, paying attention to their limb's health and conformation is vital. One of the most common diseases in the limbs of horses is laminitis. Horses with this condition suffer from lameness because it affects laminar tissue. In addition to clinical signs, radiographic criteria are essential for identifying this disease.OBJECTIVES: It is predicted that examining the effectiveness of quantitative radiographic criteria of the hoof can be helpful in the diagnosis of laminitis. Therefore, in this study, five quantitatively effective factors were investigated before and after hoof trimming to determine the changes in the radiographic diagnosis of laminitis.METHODS: A total of 11 clinically healthy horses were used in the current study. Using Marco DICOM Viewer software, lateral and dorsopalmar radiographs from the hoofs of both forelimbs were evaluated for the diagnosis of laminitis using effective quantitative criteria. Using SPSS version 24, paired T-tests were used to analyze quantitative data. P≤0.05 was considered significant.RESULTS: According to the results of this study, there were no significant differences between the right and left forelimbs after hoof trimming. On the other hand, significant differences were observed in the four following criteria: dorsal thickness between the dorsal surface of the third phalanx and the dorsal surface of the hoof, the angle between the dorsal surface of the third phalanx and the dorsal surface of the hoof, sole thickness, and the ratio of the third phalanx dorsal surface thickness to its maximum length in each forelimb before and after hoof trimming.CONCLUSIONS: During the radiographic examination, the hoof should be positioned in a standard way to diagnose laminitis accurately. However, if the hoof is not trimmed or not trimmed properly, it can interfere with laminitis diagnosis.

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 use of plant compounds and their derivatives, such as extracts and essential oils, for combating infectious agents has attracted a great deal of scientific attention. One of the active antimicrobial compounds with plant origin is carvacrol.OBJECTIVES: The present study aimed to evaluate the antibacterial activity of carvacrol alone and in combination with the antibiotic cefixime against Escherichia coli O157:H7.METHODS: The antibacterial properties of carvacrol and cefexime were evaluated by determining the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and disk diffusion method. The checkerboard assay was used to evaluate the interaction between the carvacrol and cefexime and to determine the fractional inhibitory concentration.RESULTS: The results showed that the MIC and MBC of carvacrol and cefexime against E.coli O157:H7 were 250, 250 μg/ml (MIC, MBC) and 128, 128 μg/ml (MIC, MBC), respectively. In the checkerboard test, carvacrol was found to have a synergistic interaction with antibiotic cefixime against E. coli O157:H7 (FIC index=0.5).CONCLUSIONS: Due to the significant antibacterial activity of carvacrol, the present study introduced this agent as a new antibacterial drug with a natural origin. In addition, since carvacrol significantly increased the antibacterial potential of cefixime (synergistic properties), it could be considered as an effective compound for increasing the antibacterial power of cefixime antibiotic.

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: 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.