نشریه میکروب شناسی پزشکی ایران
سال هفدهم شماره 4 (امرداد و شهریور 1402)

 Human cytomegalovirus (HCMV) has the potential for oncogenicity and has been associated with many malignancies, including prostate cancer. This study aimed to investigate the genomic prevalence of HCMV in prostate cancer tumors.

 In this case-control study, 31 patients with prostate cancer were enrolled along with 31 patients with benign prostate cancer as the control group. The presence of the HCMV genome was assessed using real-time polymerase chain reaction (PCR). The nested-PCR test was also used to detect the UL55 virus gene. Purified PCR products were sequenced using the Sanger method by a commercial service provider (Pishgam Co.), and the results were analized using Chromase software.

 The overall mean (±SD) prostate-specific antigen (PSA) value was 9 (±7.8) ng/Ml in patients with different stages of prostate cancer, and 9 (±3) ng/Ml in the control group (P=0.09). The prevalence of HCMV DNA was 19.3% (n=6) in the case group, and 6.5% (n=2) in the control group (P=0.14). The highest prevalence was observed in stage IIA (33.3%), followed by stages III (25%), and IIB (12.5%) (P=0.016). There were close phylogenetic relationships between Iranian HCMV isolates and several reference genotypes – the closest genetic relationship was observed with the KR 534207 genotype.

 The prevalence of HCMV in the prostate cancer group was markedly higher than that of the control group, although this difference was not significant. However, the higher prevalence of HCMV in prostate cancer patients compared to the control group is indicative of the possible role of HCMV in the development of prostate cancer.

 Fungal rhinosinusitis (FRS) sets in following interactivity of fungi with Sino nasal tract when immune system is subdued. Due to varied clinical presentation of FRS with inconclusive evidence of imaging, mycological evaluation helps to identify and speciate the fungi .FRS can be diagnosed with biopsy tissue and/or deep nasal swabs. This study was undertaken to know risk factors, outcome along with utility of nasal samples with emphasis on deep nasal swabs in diagnosing fungal rhinosinusitis.

 A retrospective observational study was carried out on patients with suspected fungal rhinosinusitis (invasive and non-invasive). Both COVID 19 positive and negative patients by RT-PCR were included. Brief clinical history, associated comorbidities, risk factors and outcome were noted in these cases.  Nasal swabs or tissue sample from endoscopic biopsy obtained from patients were subjected to Gram stain, KOH mount and culture on Sabouraud dextrose agar followed by slide culture for identification.

 Of the 41 cases studied for FRS, 25 were males and 16 females. Risk factors like Post COVID-19 infection was seen in 27(65.8%), diabetes in 31(75.6%) and use of steroid during COVID-19 treatment in 16(39.02%) cases. Deep nasal swabs were received in 29(70.7%) and tissue obtained after debridement in 12(29.3%) cases. Rhizopus spp was the most common fungi isolated followed by Aspergillus spp.

 Both COVID and non-COVID patients were diagnosed with invasive/ non-invasive fungal rhinosinusitis from deep nasal swabs/tissue in the present study. Diabetes was an important comorbidity in non-COVID patients with FRS. Deep nasal swabs aided early diagnosis for both invasive and non-invasive FRS.

 This research aims to study the qualitative melissopalynology analysis, Glucose oxydase (GOX) activity, and the antibacterial effect of honey samples from different botanical and geographical origins.

 Five natural honey samples were collected from local beekeepers. The antibacterial effect was carried out towards pathogenic bacteria including Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. The antibacterial activity of honey samples was evaluated using six dilutions (80%, 40%, 20%, 10%, 5%, and 2.5%). The bacterial susceptibility to honey was evaluated by the agar well diffusion and broth dilution assays. GOX activity was determined using hydrogen peroxide as a standard with peroxydase and o-dianisidine.

 The results showed that two honey samples could be classified as monofloral (H1: Hedysarum coronarium, H4: Ziziphus sp), whereas honey samples H2, H3, and H5 are multifloral honey. All honey exhibit a good bactericidal effect against pathogenic bacteria. Statistical analysis showed that there was a correlation between GOX level and the antibacterial of honey samples.

 It can be concluded that honey is an important natural product, which has an antibacterial effect towards pathogenic bacteria. Therefore, honey should be used as an alternative therapy for the treatment of wound infected by multidrug resistant bacteria. Further research should be conducted to evaluate the mechanism and the bioactive compounds in honey.

 Helicobacter pylori is found in the stomach and the oral cavity, which plays a significant role in oral diseases and recurrent gastric infections. This study aimed to detect the presence of oral H. pylori and investigate its potential association with dental caries and erosion.

 Saliva and plaque samples were collected from two groups: a study group of 40 H. pylori-infected patients, confirmed by immunological tests, and a control group included 40 subjects who were given negative results after laboratory examination. Molecular detection of H. pylori was performed using the polymerase chain reaction (PCR). Additionally, clinical assessments of dental caries and erosions were conducted.

 The study group showed a higher prevalence of H. pylori in saliva (62.5%) compared to dental plaque (45.0%). Among the study group, 70% tested positive for H. pylori by PCR, while 30% tested negative. Dental caries experience (DMFS) was slightly higher in the study group compared to the control group, and significant differences in (DS) and (DMFT) (p<0.05). The prevalence of dental erosion was also higher in the study group compared to the control group.

 The presence of H. pylori in the oral cavity represents an important risk factor for dental caries and erosion.

 The treatment of bacterial infections especially of the family Staphylococcaceae is a major health burden that has led to economic losses, high morbidity and mortality rates globally. This study aimed at isolation and characterization of coagulase positive, methicillin and multi-drug resistant Staphylococci isolated from wounds of patients.

 Forty-five wound swabs were collected using sterile swab sticks. Isolation was done by streaking technique on mannitol salt agar. The isolates obtained were screened for methicillin and multi-drug resistance using disk diffusion method. Biochemical and molecular characteristics using 16S rDNA gene were used to identify the selected isolates.

 Four Staphylococci isolates resistant to methicillin (0.00 mm to 9.0 mm) were selected out of thirty-one. Reactions to catalase and coagulase productions were positive. Three out of the four isolates were identified using their 16S rDNA genes and they were found to be closely related to other family members of Staphylococcaceae at GenBank. The fourth isolate was identified using its biochemical characteristics as Staphylococcus sp. HFS4. The three identified isolates were M. sciuri HFS1 (ON340756), M. sciuri HFS3 (ON340770) and S. haemolyticus HFS2 (ON35435). The four isolates were resistant to more than five antibiotics that cut across all the classes of antibiotics. The multiple antimicrobial indices were between 0.5 and 0.8.

 There is need for regular antimicrobial resistance surveillance within the hospital environment, earlier detection and correct prescription of potent antimicrobials will check the spread of Staphylococci infections and their virulent genes.

 Dengue virus infection remains a health problem. Dengue Virus Non-Structural protein 1 (NS1) increases the release of proinflammatory cytokines that induce intestinal zonulin expression. As a result, the ZO-1 protein translocates to the cytoplasm, which increases enterocyte permeability. This study aimed to investigate the effects of dengue NS1 on intestinal zonulin and ZO-1 expression, intestinal weight, and serum LPS.

 This experiment used 18 ddY mice with a pre-posttest control group design. Mice were randomly divided into control (C), PBS (T1), and PBS+NS1 (T2) groups. Mice in the T1 and 2 groups were intravenously injected with 500 µL PBS and 50 µg NS1, respectively. Blood samples were collected before and after the three-day treatment. The intestines were weighted directly and were then embedded with formalin for immunostaining. Serum LPS were determined using ELISA. Data were analyzed using paired t-test and ANOVA.

 The T2 group had the highest zonulin expression (histoscore=8) compared to the T1 (histoscores=7.33) and C (histoscore=6.33) groups but were not significant (p=0.135). Intestinal ZO-1 expression did not increase in the T groups compared to the C group (p=0.368). The intestine weight in the C group was significantly lower than the T group (p=0.001). After treatment, serum LPS levels in the T2 group were higher (0.34 pg/mL) than before treatment (0.12 pg/mL; p=0.118), but not in the T1 group (p=0.384). Meanwhile, there was a significant decrease in serum LPS levels in the C group (p=0.046).

 Injection of 50 µg dengue virus NS1 has a minor effect on intestinal permeability in ddY mice.

 Lactobacilli are known for their probiotic activities and potential to combat pathogens. This study aimed to investigate the antimicrobial activity of Lactobacillus strains isolated from various sources against pathogenic bacteria, with the objective of identifying potential bacteriocin producers.

 A total of 140 samples were collected from different sources. Samples were cultured on selective media such as MRS and SL agar for isolation of Lactobacillus spp under anaerobic conditions. The suspected bacteria appeared Gram positive, diploid bacilli or coccobacilli. The isolates were subjected to various biochemical tests such as catalase, oxidase, also hydrolysis test was done by growing on MRS containing 1% calcium carbonate. The antimicrobial activity of bacteriocin was screened by using three method including (Agar-plug diffusion method, Filter paper disc method and agar-wells diffusion method). Molecular identification of the best bacteriocin producer was achieved using 16S rRNA sequencing.

 The study found that L. helveticus DF isolated from feces exhibited the highest bacteriocin production efficiency. Concentrated bacteriocin from L. helveticus DF demonstrated potent antibacterial activity against multi-drug resistant bacteria, with 80 AU/ml against Pseudomonas aeruginosa and 40 AU/ml against Enterococcus faecalis.

 The study highlights the potential of Lactobacillus bacteriocins as effective antibacterial agents, offering a promising alternative to combat antibiotic resistance. Further research is warranted to elucidate the mechanisms and assess the safety and efficacy of bacteriocins for their use as bio-therapeutic agents.

 Blastocystis spp. is an anaerobic, single-celled intestinal parasite with global prevalence that is found in the lower digestive tract of humans and many animals, such as numerous types of vertebrates. To date, 17 genotypes have been identified. Nine of these genotypes are common in humans. Organism is found in both healthy and compromised individuals, hence its pathogenicity is controversial. The aim of this study was to investigate the frequency of Blastocystis spp. genotypes in Bandar Abbas, Iran during 2022.

 Totally, 378 stool samples were collected from hospitals and health centers of Bandar Abbas, Hormozgan province, Iran. Stool samples examined by direct and formalin ethyl acetate (concentration) methods. Positive samples for Blastocystis spp. isolated and DNA extraction was done. All extracted DNA specimens analyzed by nanoDrop to ensure about quantity and quality. The PCR technique was used to identify parasite genotypes and all samples were sequenced for genotype confirmation.

 Finally 14 samples detected as positive microscopically and 8 samples confirmed by PCR. Identified isolated genotypes were 4 samples ST1 (50%), 3 ST3 (37.5%), and one ST2 (12.5%). All negative and positive samples analyzed 3 times to confirm accuracy. 

 Blastocystis sp. is the most common protozoan parasite of the human intestine, which despite the studies, there are many uncertainties about its pathogenicity.  The current study was designed and carried out during the Covid-19 pandemic and lower positive patients may be related to severe hygiene considerations and consumption of various antibiotics amongst the population.

 Nowadays, non-albicans Candida are common in human pathogens, and some of these cases were found in milk. Therefore, as well as the lack of accurate estimates of its global prevalence and severity, the present study aims to assess the demographic features of non-albicans Candida (NAC) spp. and determine the species distribution of NAC. It was also evaluating the in vitro Azole susceptibility of NAC species and identified the erg11 gene and erg11 expression in fluconazole-resistant isolates of NAC spp., in Iran.

 In the present study, non-albicans Candida, including Candida glabrata, Candia krusei, Candida parapsilosis, and Candida tropicalis, were isolated and identified from 14 farms (raw milk) and human patients using culture methods, Real-Time PCR and sequencing. The resistance and susceptibility of the samples to azole were examined and erg11 expression was evaluated by RT-qPCR. The results were analyzed by REST Software to compare the levels of erg11 gene expression involved in drug resistance of NAC.

 74 and 52 NAC strains were isolated in 262 collected milk samples and human samples. Based on ITS sequencing, 0.76% were identified as C. glabrata, 2.29% C. tropicalis, 4.19% C. parapsilosis, and 19.8% C. krusei. The expression of erg11 gene in the NAC was increased in samples isolated from humans compared to samples isolated from livestock (P>0.05), while no significant difference was found in the case of Candida glabrata isolated from both sources (P<0.05). All NAC isolates were sensitive to flucytosine.

 NAC isolates from cows' milk have antifungal resistance genes while they had not taken any antifungal drugs. The resistance gene is transferred from antifungal agents in crop protection medications. Clinical isolates also had increased resistance to antifungal activity. Also, using Azole antibiotics can increase resistance gene level activity. This phenomenon should be considered for treatment program protocols.

 Human papillomavirus (HPV) is a widespread sexually transmitted infection (STI) with significant health implications. This study aimed to assess the reactivity of TGF, IL10, and MCP-1 with Leukotriene B4 in women diagnosed with cervical HPV infections. Additionally, we considered the histological grade and clinical stage of the patients.

 In this observational study, a total of 50 cervical infection samples from women diagnosed with HPV and 50 samples from healthy controls were collected between January 2nd and August 1st, 2022, from attendees of Al-Elwea Hospital for Delivery and Children. IgM and IgG antibodies against HPV antigens were detected in patient sera. Polymerase chain reaction (PCR) was employed to amplify the IL-6 gene sequences of lengths 249 and 431 base pairs. Interleukin levels were quantified using the enzyme-linked immunosorbent assay (ELISA) technique. HPV infection diagnosis was confirmed using cervical cytology (Pap smear) and HPV DNA testing before study inclusion.

 The average age of the patients was [35.02±10.73]. The findings revealed that among papilloma cases, 30 (85.7%) exhibited normal levels of both IgM and IgG, while 5 cases (14.3%) showed normal IgM levels and elevated IgG levels. Additionally, 10 cases (66.7%) of papilloma demonstrated increased IgM levels and normal IgG levels, in contrast to 5 cases (33.3%) with elevated levels of both IgM and IgG. Importantly, a positive correlation was observed between IgM and IgG levels and the concentrations of TGF, MCP-1, IL-10, and LTB-4 (r=0.896**, 0.678**, 0.692**, 0.894**), (r=0.894**, 0.546**, 0.570**, 0.618**), respectively.

 The study revealed a significant positive correlation between IgM and IgG levels and the concentrations of TGF, MCP-1, IL-10, and LTB-4, indicating potential immunological interactions in HPV infection. High Ct values for both patients and controls, along with HR-HPV gene expression, suggest a proportional relationship between gene concentration and Ct value.

 Escherichia coli  (E.coli) is an essential bacterial indicator in the control of pharmaceutical quality and other similar fields. There are some biosensors designed for its detection based on electrochemical transduction methods. A biosensor with reduced graphene oxide was modified. Traditional methods are time-consuming and high prices, so in this study a new biosensor with modification of reduced graphene oxide (rGO) as a kind of carbon composition on glassy carbon electrode (GCE) with Au Nanoparticles (Au NPs) decoration was used for E. coli detection.

 Reduced graphene oxide (rGO)  modified on glassy carbon electrode (GCE) and chronoamperometric and reduction methods were used for Au NPs decoration and it was completed with polyclonal E. coli antibody and 0.5 W/V% Bovine Serum Albumin (BSA) solution. Morphology and structure of rGO and Au NPs in GCE/rGO/Au NPs/ E. coli polyclonal antibody/ BSA biosensor were verified by SEM (Scanning Electron Microscope) during modification steps. E. coli detection in dissimilar samples was performed by Square-Wave Voltammetry (SWV) and Cyclic Voltammetry (CV) techniques which were set in 0.1M phosphate buffer solution (PBS) (pH 7.4) mixed with 0.5mM acetaminophen. In comparison with the biosensor, the classical method of detection was performed with serial dilutions of E. coli ATCC 8739 (1×101–1×108 CFU/ml) which was cultured on media. 

 Despite the two methods used to stabilize Au NPs, scanning electron microscope (SEM) images showed that the current difference did not increase due to gold particles. Its modification had not significant change in current and it was not a successful experiment for E.coli detection.

 Compared with the plate method, biosensor couldn’t substitute with conventional methods for E. coli detection.

 Immune response to BNT162b2 COVID-19 vaccine among people is not the same. Genetic polymorphism in inhibitory immune checkpoints may have an important role in immune response after vaccination; therefore, we aimed to demonstrate the role of CTLA-4 rs733618 gene polymorphism and the amount of circulating CTLA-4 in individuals vaccinated with the BNT162b2 vaccine following the booster dose.

 This research is a cross-sectional study performed on 180 healthy adults (above 18 years old) vaccinated with the BNT162b2 COVID-19 vaccine 21-30 days after the second dose at the community dwelling from December 2021 to April 2022. After DNA extraction from the sample’s blood, Allele-specific polymerase chain reaction (ASPCR) was developed to detect single nucleotide polymorphisms of CTLA-4 rs733618. The levels of IgG in the serum, which are directed towards the spike protein-1 and soluble CTLA-4, were quantified using an Enzyme-linked Immunosorbent Assay (ELISA).

 The current study showed no significant association in CTLA-4 rs733618 genotype distribution and immune response to the BNT162b2 vaccine, Furthermore, there was a highly significant difference between CTLA-4 serum levels and CTLA-4 rs733618 genotype frequency.

 CTLA-4 -1722 T/C rs733618 is not significantly related to immunological response to the BNT162b2 vaccine. The level of s-CTLA-4 production may be affected by CTLA-4rs733618 polymorphism. rs733618 (T/C) and rs733618(C/C) genotypes significantly related with high level of s-CTLA-4, while rs733618 T/T genotype related with low level of s-CTLA-4.

 Antimicrobial resistant (AMR) is a serious global threat and World Health Organization (WHO) declared AMR is the one among 10 public health threat facing the humanity. Increase in Carbapenem resistant is the major problem which hinders the patient treatment. Various genes are responsible for carbapenem resistant but the gene bla-NDM has the ability to inactivate various carbapenem antibiotics namely meropenem and imipenem etc. Therefore, this study was aimed to observe the prevalence of bla-NDM gene in the carbapenem resistant Klebsiella spp isolates.

 The isolates of multidrug resistant Klebsiella spp were collected from the central diagnostic laboratory, Sree Balaji Medical College and Hospital, India after obtaining the ethical clearance. The isolates were screened for the carbapenem resistant using Rapidec Carba NP method. The presence of bla-NDM was confirmed using conventional PCR technique.

 Most of the isolates showed resistant to ampicillin, gentamycin, cefazolin, imipenem and meropenem. 45% of the isolates of K.pneumonia showed resistant to carbapenem group of antibiotics and 20% of the K.oxytoca showed carbapenem resistant. Among the carbapenem resistant 23.75% of the K.pneumonia showed the presence of bla-NDM gene and 6.6% of K.oxytoca showed the presence of bla-NDM gene. Continuous increasing in the carbapenem drug resistant organism is worrisome and the judicious usage of antibiotics should be practised.

 Rheumatic Fever (RF) is a non-suppurative systemic inflammatory disease with a "delayed autoimmune" caused by Group A streptococcal infections. Inadequate RF treatment causes recurrent RF which is a predisposing factor for Infective Endocarditis.

 A 39-year-old woman was admitted to the hospital with chief complaints of intermittent fever since one month ago, shortness of breath during strenuous activities, weight loss, and easy fatigue. Heart examination palpable thrill, grade 3/6 systolic murmur at apex. Laboratory results were Hb 9.4 g/dl, ESR I: 61 mm, ESR II: 159 mm, ASTO 2563 IU/ml, and CRP 8.9 mg/l. Results of Transthoracic echocardiography revealed vegetation on the posterior mitral leaflet measuring 2.09 x 0.7 and blood culture identified Streptococcus alactolyticus.

 Patients diagnosed with recurrent rheumatic fever and infective endocarditis received empirical antibiotic therapy for 2 weeks and corticosteroids, after follow-up the patient experienced clinical improvement and echocardiography showed reduced vegetation size.