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Effects of Cadmium Poisoning on the Expression of NF-kB/MAPK and CD163 Genes in Rat Adrenal Gland
2023
Gholami, Atieh | Marjani, Zahra | Hesaraki, Saeed
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.
Показать больше [+] Меньше [-]Comparison to Methods; Serum Antibody ELISA and Fecal Nested-PCR to Diagnose Mycobacterium avium Subspecies Paratuberculosis Subspecies Infection in Cattle
2023
Kolivand, Ali | Haji Hajikolaei, Mohammad Rahim | Nouri, Mohammad | Khosravi, Mohammad | Gharibi, Dariush
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.
Показать больше [+] Меньше [-]Radiographic Evaluation of Effective Quantitative Criteria in Diagnosis of Laminitis before and after Trimming in Healthy Horses
2023
Soroori, Sarang | Tavakoli, Amir | Akbarein, Hesameddin | Bonyadi, Mojtaba | Shateri Amiri, Banafshe
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.
Показать больше [+] Меньше [-]A Comparison Survey on Native and Denaturation Conditions for Solubilization and Purification of Toxocara canis C-type Lectin Recombinant Protein
2023
Malekzadeh, Parmida | Hosseini, Seyed Hossein | Jalousian, Fateme | Akrami, Mohammad | Amini Nia, Narges
BACKGROUND: Toxocara canis C-type lectin (T. canis-CTL) is the main protein part of the secretory-excretory product secreted by Toxocara canis infective larvae. T. canis-CTL can stimulate immune response-mediated regulatory T lymphocytes, increase the FOXP3+ cells population, and reduce severe inflammatory responses. T. canis-CTL is a promising candidate for immune modulation in some autoimmune diseases, deserving further investigation.OBJECTIVES: The current research aimed to purify the recombinant T. canis-CTL under denaturation and native condition to increase exploration and maintain biological activity.METHODS: The expression vector, pET32a was constructed with the partial sequence 660bp of T. canis-CTL and expressed in E. coli BL21 (DE3). T. canis-CTL protein expression was induced by IPTG (1 mM) at 37°C after 6 h. In this study, different buffers were used for cell explosion and recombinant protein solubilization, including lysis buffer with urea (8 M, pH=8) and lysozyme enzyme as well as lysis buffer with Imidazole (0.01 M) and lysozyme enzyme, and previous buffers in addition to sonication. The effect of these buffers was evaluated in bacterial cells explosion, using Gram-staining and microscopic examination. Recombinant T. canis-CTL protein was extracted and purified under denaturation and native condition using Ni-NTA affinity chromatography by agarose and sepharose resin. A New Zealand male rabbit was immunized with the recombinant protein to evaluate the bioactivity of the protein. RESULTS: Lysis buffer with urea (8 M, pH=8) and lysozyme enzyme, in addition to sonication, provided acceptable results, and an additional amount of recombinant T. canis-CTL protein was secreted in the buffer. Protein purification under denaturation conditions with Ni-NTA agarose affinity chromatography also provides further recombinant protein. Most of the induction of recombinant T. canis-CTL with 41 KDa molecular weight was collected 6 h after induction at 37°C. Dot-blot results illustrate the brown dot, which showed a 1:500 titer of specific IgG polyclonal antibody has developed in the sera of rabbits immunized with T. canis-CTL recombinant protein.CONCLUSIONS: The denaturation condition did not affect the biological activity of the T. canis-CTL recombinant protein and can recover a further amount of recombinant proteins.
Показать больше [+] Меньше [-]Measuring Mercury Level in Edible Tissue of Farmed Whiteleg Shrimp (Litopenaeus vannamei) in Bushehr Province of Iran Using Cold Vapor Method
2023
Sadeghi Limanjoob, Reza | Zargar, Ashkan | Taheri Mirghaed, Ali
BACKGROUND: Many toxic elements enter human food in different ways by various industries which can put people's lives in danger. Heavy metals can be rarely removed from the body after absorption and deposition in tissues, which can lead to diseases and complications in the body. Mercury is one of the heavy metals that can posion people after consumption of contaminated seafood. The measurement of pollutants such as mercury that present in aquatic animals and environment is one of challenges for humans.OBJECTIVES: This study aims to measure the amount of mercury accumulation in the edible tissue of whiteleg shrimp (Litopenaeus vannamei) found in farms of Bushehr Province in Iran.METHODS: In this research, 70 whiteleg shrimps were collected during four sampling stages in July, August, September and October for two consecutive years. Total mercury level was measured by the cold vapor method.RESULTS: The level of mercury was 0-0.009 mg/kg of body weight, while the recommended limit for mercury is 0.1 mg/kg according to the WHO. The microscopic study on tissue sections did not show any histopathological changes.CONCLUSIONS: The mercury level in the edible tissue of whiteleg shrimps in Bushehr province is much lower than the recommded level and does not pose any danger to residents and consumers.
Показать больше [+] Меньше [-]The Antioxidant and Anti-Inflammatory Effects of Ischemia Post-Conditioning on the Injury Induced by Ischemia/Reperfusion in the Rat Liver
2023
Ghasemi Pour Afshar, Neda | Arab, Hossein Ali | Vatannejad, Akram | Ashabi, Ghorbangol | Golabchifar, Ali Akbar
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.
Показать больше [+] Меньше [-]Effect of Gamma Irradiated Saffron Petal Ethanolic Extract on Viability of Lacticaseibacillus paracasei M4PM99 and Assessing their Antioxidant Properties in Probiotic Yogurt
2023
Ghorbanzadeh, Pouria | Parsaeimehr, Mahnoosh | Heidarieh, Marzieh | Jebellijavan, Ashkan
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.
Показать больше [+] Меньше [-]Consumption Pattern of Local Dairy Products and the Related Factors in Female Residents of Qom Province, Iran in 2022: A Community-Based Cross-Sectional Study
2023
Derakhshani, Maryam | Keshvari-Shoja, Zeinab | Aligol, Mohammad | Ahangari, Roghayyeh | Mohammadbeigi, Abolfazl
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.
Показать больше [+] Меньше [-]Antibiotic Resistance in Pathogenic Bacteria, the Causative Agents of Bacterial Diseases in Farmed Rainbow Trout (Onchorhynchus mykiss) in Iran
2023
Soltani, Mahdi | Rakhshanimehr, Kambiz | Mirzargar, Seyed Saeed | Zargar, Ashkan | Shohreh, Poulin | Asadi, Sepideh
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.
Показать больше [+] Меньше [-]Evaluating the Effect of Culture Supernatant of Pseudomonas aeruginosa on Removing the Inhibitory Effect of Heparin in Real-Time PCR Test
2023
Ashrafi, Aysan | Staji, Hamid | Keramati, Keyvan
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.
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