فصلنامه پاتوبیولوژی مقایسه ای
سال هفدهم شماره 68 (تابستان 1399)

In this study, detection and recognition of Staphylococcus aureus enterotoxin was performed by designing a Dot ELISA sensitive kit. In the present study, nitrocellulose membrane (NCM) with 0.22 micron pores was applied as the bed, and 1:500 diluted specific antibody of Staphylococcus aureus enterotoxin was coated on the NCM. Next, The HRP-tagged antibody was added with dilution of 1/2000, and the substrate was added gradually and rewashed. In this experiment, the samples were manually contaminated and then compared with the ELISA sandwich method as a gold standard. The kit is capable of detecting 50 ng/ml and higher of toxin A, and if the amount of toxin is less, it will report it negatively. In this test, of which 40 samples were collected and 4 positive cases were reported, it was revealed that about 10% of the collected samples were contaminated. Nowadays, using the Dot ELISA kit as a quick method with relative sensitivity and properties, it is possible to improve the repeatability of detecting Staphylococcus enterotoxin A in creamy candies.

Cancer is the second factor of death in world on the basis of WHO reports on 2018. Application of fungal polysaccharides is one of the cheaper, less dangerous, less side effects, and newer in the treatment and prophylaxis of cancer. Object of this research is isolation and identification of endemic fusarium fungus, its glucan extraction, determination of chemical characteristics by HPLCand FT-IR and anticancer effects on cell lines of LCL, Hela.In this study fusarium fungus was isolated from soil of karaj district and identified on the basis of microscopic and macroscopic and genetical characteristics as a fusarium genus .Then fungus was grown into selective broth medium for obtaining of the most biomass for extraction of glucan .Glucan of fungus was extracted by boiling water method.Different amounts of extracted glucan were treated to the cell cultures of lcl and Hela and its cytotoxicity effects were surveyed.Results showed that glucan polysaccharide had anticancer effects against cell lines of LCL and no anticancer effects against Hela cell lines. Cytotoxicity effects of glucan showed by colorimetry MTT method.

Epidermophyton floccosum is one of the worldwide anthropophilic dermatophytes that invade keratinized structures such as hair, skin, and nail, causing dermatophytosis by secreting important proteases such as subtilisin. This study aimed to evaluate the presence of SUB3 and SUB6 encoding serine proteases in the isolate, which received from the fungi culture collection of Tehran University of Medical Sciences, Faculty of Public Health. Special primers designed according to the highly conserved regions of similar genes in other dermatophytes. Genomic DNA and designed primers used in PCR, then PCR products sequenced with ABI PRISM®3730XL automated Sanger sequencer, and presence of 2 new subtilisin genes (SUB3 and SUB6) were confirmed and recorded in NCBI(with the accession numbers MN206114, MN177931 respectively). The coding sequence of SUB3 found to contain 861 nucleotides, which encodes a polypeptide with 287 amino acids. The coding sequence of SUB6 found to contain 699 nucleotides that encode a polypeptide with 233amino acids. Comparing the sequences with Gene Bank database information, revealed significant homology with other dermatophytes. Achieving a better understanding of the molecular characteristics of virulence genes may help develop effective therapies and prevention strategies.

The current study was conducted to investigate the effects of clove (Syzygium aromaticum L.) essential oil on hepatiorenal histopathology and biofunction, protein profile and immunity in broiler chickens. A 42-day fully randomized trial was conducted using 180 male broilers (Ross 308) divided into 4 main groups (15 birds in each) with three replicates, supplemented with the different levels of herbal essential oil as follow: I) Control (basal diet only); II) basal diet + 250 ppm oil; III) basal diet + 350 ppm oil; Ⅳ) basal diet + 450 ppm oil. The following indices were evaluated: relative weights of liver and kidney, renal injury biomarkers including urea and creatinine, hepatic biomarkers including the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT), protein profile including total protein, albumin and globulin and finally immunoglobulins A, G and M. Furthermore, histologic sections of liver and kidney were studied using a semi quantitative method. The obtained results indicated that clove could increase the levels of proteins and immunoglobulins dose dependently. However, the highest dose (450 ppm) could significantly alter hepatic and renal histopathology and biomarkers. Moreover, pathologic damages were more severe in liver compared to kidney. In conclusion, clove essential oil exerts beneficial effects on protein profile and immunity in broilers, however, it may damage liver and kidney in higher doses. Therefore, optimal and safe doses of the plant should be used.

Theileria are important hemoprotozoan parasites of cattle, transmitted by ixodid ticks leading to diseases which range from mild in apparent reactions to highly fatal diseases. This study aimed to determine the variety of Theileria species among cattle of Shahreza city, the south part of Isfahan Province. A total of 253 blood samples were randomly collected via the ear vein and jugular vein from healthy cattle for microscopy and molecular examination, respectively. The extracted DNA from blood cells was amplified by Theileria-all primers, which amplify an approximately 400bp DNA fragment from the region of the 18S rRNA gene from various members of the genus Theileria. All cattle positive samples were further analysed for the presence of Theileria annulata and Theileria orientalis by specific semi-nested PCR. T. annulata and T. orientalis were identified by specific semi-nested PCR in 54.9% and 47% of cattle blood samples, respectively. Chi-square (x2) tests were used to compare molecular prevalence values relative to Season, Farm, Type, Hygiene, Vectors, Age, and Milk Yield. T. orientalis prevalence was significantly higher in cattle in summer as compared to those in other seasons (p < 0.05). The prevalence rates of T. orientalis in different seasons, hygiene levels, and vectors were significantly different (p < 0.05). T. orientalis prevalence was higher in spring, farms with tick vectors and normal level of Hygiene. The examination of 50 microscopic fields showed 14.39% sensitivity and 99.12% specificity compared to molecular examination. The Kappa coefficient between molecular and microscopy (50 fields) techniques indicated a poor level of agreement (Kappa= 0.124). This study is the first molecular detection of Theileria species from cattle in the south of Isfahan province, Iran.

Pecten is a fully vascularized and pigmented structure of the eye in avian species which is probably involved in retinal nourishment. The structural features of pecten can be somewhat different in different bird species. With the aim of investigating the histological and anatomical structure of pecten in domestic Iranian breed of chickens, 8 chickens (4 hens and 4roosters) were purchased from Chicken Breeding Institute of Mazandaran and after slaughter, the eye balls were removed and fixed in 10% buffered formalin for anatomical and histological evaluations. It was observed that the pecten structure was consisted of 20 lamellae with different heights in both eyes of both sexes. The basal width of lamellae was 0.51±6235 micrometer in roosters and 0.47±6207 micrometer in hens. The heights of lamellae were different in each pecten oculi. Blood sinuses were present at the attachment site of pecten oculi to retina. In conclusion, the number of lamellae of pecten oculi in domestic Iranian breed of chickens is more similar to quails and turkeys while the size of their pecten oculi is close to galah parrots and domestic chickens with histological features similar to other species.

Due to universal vaccination program, the prevalence of many dangerous diseases among infants and adults has been significantly reduced. In order to obtain appropriate strategies for regulating immune responses that lead to increased vaccine efficacy, here we investigated the pattern of immune response following diphtheria-tetanus vaccine administration in mice model. Diphtheria (D) and tetanus (T) monovalent and divalent (DT) vaccines have been separately formulated in the presence and absence of adjuvant (aluminum phosphate gel). Each vaccine has been injected for 5 times in Balb/c mice.  Two weeks after the last injection, the spleen of the mice was removed and the spleen cells were exposed to the selected antigens and the levels of Interleukin 4 (IL-4) and IFN- ɤ (Interferon gamma) were measured in cell culture supernatant. According to our results, the levels of IL-4 was significantly increased in mice receiving adjuvant vaccines compared with the non-adjuvant and control groups, while this increase was not significant for IFN-Ƴ. This results mentioned that the orientation of immune responses is more towards T helper 2 responses (Th2), which could be related to the nature of the antigen, the vaccine formulation strategy and the percentage of antigen uptake in the alum gel.