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JoMMID Journal

Pasteur Institute of Iran, Medical Microbiology and Infectious Diseases, Faculty Member

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Journal of Medical Microbiology and Infectious Diseases (JoMMID) is an international open-access peer-reviewed journal in English that publishes original research papers, review papers, and clinical studies related to infectious diseases of bacterial, viral and parasitic origin. The journal also accepts papers describing research on epidemiology of infection, diagnosis and treatment, pathogenesis, antibiotics and resistance, and immunology.
JoMMID accepts original articles, review articles, short communications reporting clinical and basic research studies, case studies and clinical trials of significant scientific and ethical standards in the following areas:
- Infectious diseases and public health;
- Emerging and reemerging infectious diseases;
- Laboratory diagnosis of human and animal infections;
- Antimicrobial resistance and treatment protocols;
- Molecular epidemiology of pathogenic agents;
- Host-pathogen interactions and susceptibility factors;
- Immune responses, deficiencies and vaccine candidates;
- Microbial pathogenesis;
- Vector-borne Diseases.

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Papers by JoMMID Journal

Ophidascaris sp. in an African Rock Python (Python sebae) in Ghana: A Case Report

by JoMMID Journal and L. Bimi

Journal of Medical Microbiology and Infectious Diseases, 2021

This study reports an Ophidascaris sp. infection in the gastrointestinal tract of a python snake ... more This study reports an Ophidascaris sp. infection in the gastrointestinal tract of a python snake found at a construction site in Labone, a suburb of Accra and the capital city of Ghana. Examination of the rectal contents of the snake by the zinc sulfate centrifugal floatation method revealed helminths eggs. Besides, examining various sections of the alimentary canal revealed adult worms of the genus Ophidascaris in the anterior half of the small intestine just below the stomach's pyloric sphincter. Also, milking one of the female worms onto a microscope slide and examining the exudate by microscopy following staining with Lugol's iodine dye revealed eggs. Our findings may alert the possibility of zoonotic transmission of the parasite from pythons to humans, especially in the suburban/rural areas where people consume snake meat.

Frequency of cbrA, cbrB, ndvB, and phoBR Genes in Relation to Biofilm Formation in Pseudomonas aeruginosa Clinical Isolates

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: After Staphylococcus aureus and Escherichia coli, Pseudomonas aeruginosa is the thi... more Introduction: After Staphylococcus aureus and Escherichia coli, Pseudomonas aeruginosa is the third cause of hospital-acquired infection (HAI). This bacteria's ability to colonize in different environments, especially in hospitals and biofilm formation, has added to its impact as an HAI. The molecular mechanism of biofilm formation is not well understood, but several genes contribute to this phenomenon. This study investigates the frequency of cbrA, cbrB, phoBR, and ndvB genes in biofilm-forming P. aeruginosa isolates. Methods: Fifty P. aeruginosa clinical isolates were collected from various sources such as urine, ulcer, blood, secretions, and trachea in Milad Hospital, Tehran, from 2017 to 2018. Biofilm formation in the isolates was assessed by the microtiter plate assay, and the frequency of cbrA, cbrB, phoBR, and ndvB genes was investigated by PCR. Results: Among the 50 isolates, 44% were strong biofilm former, 34% moderate biofilm former, 12% weak biofilm former, and 10% did not form biofilms. PCR revealed a frequency of 94% for the cbrA gene, 78% for cbrB, 96% for ndvB, and 48% for phoBR. The coexistence of all four genes was 68% in strong biofilm former isolates, 41% in moderate biofilm former isolates, 37% in weak biofilm former, and zero in the isolates that formed no biofilm. Conclusion: The high frequency of ndvB and cbrA genes and the coexistence of ndvB and cbrB suggest the contribution of these genes in the biofilm formation of P. aeruginosa.

In-silico Immunomodelling of SARS-CoV-2

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense si... more Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense single-strand RNA virus belonging to the Coronaviridae family, responsible for coronavirus infectious disease 2019 (COVID-19) with the rapid transmission. This study aimed to characterize and compare SARS-CoV-2 and SARS-CoV major viral proteins and predict antigen proteasomal cleavage patterns, MHC class I processing and presentation, and B T-cell and anti-inflammatory epitopes. Methods: The amino acid sequences of spike surface (S) glycoprotein, membrane (M) glycoprotein, envelop (E) protein, and nucleocapsid (N) phosphoprotein was obtained from NCBI. The sequences were aligned by MEGA 7.0 and modeled by SWISS-MODEL. The proteasomal cleavage pattern, MHC class I processing, and T-cell epitopes were predicted via IEDB analysis and EPISOFT. The B-cell epitopes were predicted by BepiPred 2.0. Also, the prediction of anti-inflammatory epitopes was performed by AntiInflam. Results: Two major antigen proteins, S glycoprotein and M glycoprotein of SARS-CoV-2, respectively, showed 26.57% and 20.59% less efficiency in proteasomal cleavage and presentation to MHC class I, comparing SARS-CoV. There were fewer B-cell predicted epitopes in SARS-CoV-2, comparing SARS-CoV. The anti-inflammatory properties of SARS-CoV-2 S glycoprotein and N protein were higher than SARS-CoV. Conclusion: It seems that the evolution of SARS-CoV-2 is on the way to reducing antigen-presenting to MHC class I and escaping cellular immunity. Moreover, the predicted hotspot epitopes potentially can be used to induce adaptive cellular immunity against SARS-CoV-2. Besides, SARS-CoV-2 appears to be less immunopathogenic than SARS-CoV due to its higher anti-inflammatory proteins.

Comparison of PCR with Serology for Detecting Acute Hepatitis A Virus Infection

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Early detection of acute Hepatitis A virus infection (HAV) allows adopting proper t... more Introduction: Early detection of acute Hepatitis A virus infection (HAV) allows adopting proper treatment measures, rapid recovery, and avoiding side effects. This study compares PCR assay with serology for diagnosing acute HAV infection. Methods: Twenty samples from patients presenting clinical symptoms of acute hepatitis were tested for anti-HAV IgM antibodies. Genomic RNA was extracted from IgM-positive samples, cDNA was synthesized and examined for genomic HAV using a specific HAV real-time detection kit and a nested PCR. Results: Among 20 sera, 14 were positive for anti-HAV IgM antibodies. The specific real-time PCR and nested PCR showed agreement, and both detected HAV genetic material in 3 out of 14 samples. Conclusion: High levels of anti-HAV IgM antibodies do not necessarily indicate acute HAV infection in people presenting clinical symptoms of the disease. Measuring IgM antibody levels alongside molecular detection of virus genome by DNA-based methods assay can lead to an accurate, timely, and reliable diagnosis of active HAV infection.

The Effects of Dehydrozingerone on Growth, Biofilm Formation, and Ergosterol Biosynthesis of Candida albicans

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Candida albicans can cause various diseases, which might lead to various cases of l... more Introduction: Candida albicans can cause various diseases, which might lead to various cases of life-threatening diseases. Biofilm is a specific feature of C. albicans formed on mucosal surfaces and medical devices. Moreover, biofilm protects Candida cells from antifungals and makes the treatment challenging. Here, we studied the effects of dehydrozingerone on C. albicans growth, ergosterol biosynthesis, biofilm formation, and the expression of an essential gene involved in yeast-hypha transition. Methods: C. albicans cells were treated with serial two-fold concentrations of dehydrozingerone (0.125-2 mg/ml) for 48 h at 35 °C. The weights of the fungal cells were estimated as a sign of fungal growth. Biofilm formation was evaluated by a tetrazolium salt (XTT) reduction assay. The expression of the HWP1 gene was assayed by real-time PCR. Results: Dehydrozingerone inhibited C. albicans growth in the range of 3.57% to 84.28%, dose-dependently. The ergosterol content of yeast cells was reduced by 50% in the highest concentration. The biofilm formation was also inhibited by more than 50% at the highest concentration. The expression of the HWP1 gene was suppressed by dehydrozingerone at different concentrations. Conclusion: Our results indicate that dehydrozingerone displayed effective activity against growth, biofilm formation, and ergosterol biosynthesis in C. albicans in vitro.

Prevalence and Antibiotic Susceptibility Profile of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates in Diabetes Patients with Foot Ulcers

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: methicillin-resistant Staphylococcus aureus (MRSA) infection is common among diabet... more Introduction: methicillin-resistant Staphylococcus aureus (MRSA) infection is common among diabetes patients with foot ulcers. This study aimed to determine MRSA isolates prevalence and antibiotic susceptibility profile in diabetic foot ulcers (DFU) patients. Methods: A total of 204 patients with diabetic foot ulcers admitted to a tertiary hospital in Abia State, Nigeria, were included in the assay. Specimens were obtained by scraping the ulcer base or the deep portion of the wound edge using a sterile curette and were promptly sent to the laboratory for culture, identification, and antibiotic susceptibility test. Results: The MRSA prevalence in DFU patients was 22.1% (n=45). Male patients with DFU were more infected with MRSA (n= 26, 12.7%) than females (n=19, 9.3 %), but the difference was not statistically significant (P < 0.14). The age group 41-60 years had the highest prevalence (n=27, 13.2%), statistically significant (P < 0.02). Farmers had the highest prevalence of 9.8% (n=20) while the least (0.5%) was seen in housewives (n=1) with no statistical significance (P < 0.07). The antibiotics sensitivity pattern of MRSA showed 100% sensitivity to vancomycin and chloramphenicol but 100% resistance to penicillin, ceftriaxone, and oxacillin. The multidrug-resistant index was all > 0.2. Conclusions: The prevalence of MRSA in DFU patients in a tertiary hospital in Abia State was very high, with an alarming rate of drug-resistant bacteria due to the possibility of misuse and abuse of antibiotics among the populace, which requires collaborations from all stakeholders to prevent drug resistance in the study settings.

Expression of a Novel HIV-1 Gag-Pol-Env-Nef-Rev Multi-Epitope Construct in Escherichia coli

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Recombinant subunit vaccines have been explored against various human pathogens, ho... more Introduction: Recombinant subunit vaccines have been explored against various human pathogens, however, developing an effective therapeutic toward human immunodeficiency virus (HIV) infection has been challenging. So far, several recombinant HIV-1 antigens have been produced and examined for activation of desired immune responses. This study aimed to express an HIV-1 multiepitope protein as an antigen candidate to develop a vaccine. Methods: In this study, the codon-optimized encoding sequence of the designed multi-epitope construct (Gag-Pol-Env-Nef-Rev) was synthesized and subcloned into the pET-24a (+) expression vector. Then, expression of the target antigen was evaluated in E. coli BL21 (DE3) and Rosetta strains under different conditions (temperature, optical density/ OD600, isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration, and time). Finally, the expression of the Gag-Pol-Env-Nef-Rev multi-epitope protein was confirmed using SDS-PAGE and western blot analysis. Results: The highly conserved and immunodominant T-cell epitopes of HIV-1 Gag, Pol, Env, Nef, and Rev proteins were used to prepare a novel Gag-Pol-Env-Nef-Rev multi-epitope construct. The gag-pol-env-nef-rev gene was successfully sub-cloned in pET-24a (+) vector and subsequently expressed in BL21 (DE3) E. coli strain under optimized conditions (1 mM IPTG, 16 h post-induction, OD 600= 0.6, and 37ºC). A clear band of ~ 35 kDa was detected by western blotting using an anti-His antibody, indicating the successful expression of our target multi-epitope protein. Conclusion: Expression of the recombinant HIV-1 multi-epitope protein was optimized in a bacterial system. The expressed protein will be purified to use as a multi-epitope protein vaccine candidate in the future.

Ghana is Free from the Guinea Worm after a 33-Year Eradication Program

Journal of Medical Microbiology and Infectious Diseases, 2021

For several generations, people from some parts of Ghana have suffered in the hands of a yard-lon... more For several generations, people from some parts of Ghana have suffered in the hands of a yard-long "spaghetti-thin" worm, known as Dracunculus medinensis, which infects humans and leads to a disease known as Dracunculiasis, literally meaning "afflictions with little dragons." The disease, also commonly known as Guinea Worm Disease, Dracuntiasis, or Dracunculosis, is a 3000-year-old known parasitic infection that rarely made headlines before the mid-1980s. Guinea Worm Disease, a plague so ancient that it is believed to be the "fiery serpent torturing the Israelites in the desert," as described in the Old Testament. This paper reviews local and global efforts and strategies at eradicating the disease in Ghana and further diagnoses the problems that hindered the early realization of the desired results of these strategies. This article did not evaluate Ghana's performance in the program. It is equally arduous to unearth all the reasons contributing to the somewhat uneasy road to eradication over three decades of efforts. This review analyzes time-trends, program documents, technical and non-technical reports, and research documents that reveal that Ghana's program ended a decade of disappointing stagnation following the disruptive ethnic conflicts in the early 1990s in its most disease-endemic areas. Despite substantial reductions in the number of guinea worm cases during the mid-1980s, efforts to break the transmission chain in Ghana remained a daunting task. The efforts required continued international and political commitment, active surveillance, strengthening of interventions, and honesty of documenters at all levels.

Transient Gene Expression: an Approach for Recombinant Vaccine Production

Journal of Medical Microbiology and Infectious Diseases, 2021

The production of recombinant vaccines in green plants is an attractive and promising topic in ge... more The production of recombinant vaccines in green plants is an attractive and promising topic in genetic engineering. However, the stable transformation of green plants is a time-consuming, costly, and labor-intensive practice. Moreover, public concerns about genetically modified plants put another limitation on the development and release of transgenic plant-based recombinant vaccines. These shortcomings were addressed by developing transient gene expression systems that allow researchers to investigate candidate recombinant vaccines quickly without tedious work and high costs. A comprehensive literature review was used to gather relevant information. This approach has received much attention in various recombinant vaccine production platforms, including mammalian cell culture, insect cell culture, yeast expression systems, and, more importantly, in plant hosts. Due to their simplicity and efficiency, transient gene expression systems are now widely used to validate gene constructs and transgene expression within plant tissues. This paper describes the concept of transient gene expression and discusses the significant advantages of this approach for producing recombinant vaccines. Notably, the major types of transient gene expression viz. agroinfiltration, viral-based systems, and application of naked plasmid in plant cell culture are introduced, and some examples illustrate the pros and cons of each system. Our literature review also discusses some practical notes on the successful application of this system to provide a more comprehensive image of transient gene expression applicability in green plants. As a whole, this review contributes to the existing literature by shedding more light on various aspects of transient gene expression that have not been addressed thoroughly yet.

Protection and Immune Responses Elicited by rSAG1-PLGA Nanoparticles in C57BL/6 Against Toxoplasma gondii

by JoMMID Journal and Mojgan Allahyari

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: This study aimed to evaluate rSAG1-PLGA efficacy as a particulate vaccine in confer... more Introduction: This study aimed to evaluate rSAG1-PLGA efficacy as a particulate vaccine in conferring protection against Toxoplasma gondii infection in C57BL/6 mice. In light of our previous studies, we studied mice genotype role in eliciting immune responses by rSAG1-PLGA nanoparticles in this study. Methods: Poly (DL-lactide-co-glycolide) (PLGA) nanoparticles loaded by rSAG1 as a subunit vaccine were prepared, and C57BL/6 mice were subcutaneously immunized twice at a 3-week interval by rSAG1-PLGA, soluble rSAG1, blank PLGA, and one group kept unvaccinated. The characteristics of PLGA nanoparticles, the amounts of produced IFN-γ, IL-10, specific anti-ToxoplasmaIgGs, and the conferred protection against infection by T. gondii RH tachyzoite were assessed. Results: rSAG1-PLGA nanoparticles shared a z-average of about 450nm with negative Zeta potential. Compared with the negative control group, the mice vaccinated with rSAG1-PLGA nanoparticles produced significantly higher amounts of IFN-γ, specific anti-T. gondii IgG antibodies and higher titer of IgG2a, which resulted in longer survival times. Conclusion: The efficiency of rSAG1-PLGA nanoparticles in inducing humoral and cellular responses and consequently partial protection against acute toxoplasmosis in C57BL/6 was confirmed.

Contamination of Raw Herbs with Parasitic Protozoa and Helminths in Shushtar City, Southwestern Iran

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Intestinal parasites are among the most prevalent foodborne diseases worldwide, and... more Introduction: Intestinal parasites are among the most prevalent foodborne diseases worldwide, and raw vegetables and herbs are among the primary sources of human infection by these parasites. This study aimed to investigate the prevalence of parasitic contamination of fresh herbs in Shushtar, Khuzestan Province, Southwest of Iran. Methods: In this study, 129 herb samples from various farms were collected and washed with water. The washing waters were centrifuged, and the resulting sediments were examined by formol-ether concentration and Sheather's sugar flotation procedure, as well as a wet smear and Ziehl-Neelsen staining. Results: Among the 129 samples, 73.6% (n=95) showed contamination with at least one parasite, including trophozoite like amoebae (52.6%), followed by Giardia lamblia (14.7%), Cryptosporidium spp. (2.1%), Blastocystis sp. (21%), free-living nematodes larvae (3.1%), Trichostrongilid nematodes (1.05%), Ascaris lumericoids eggs (2.1%), Hymenolepis spp. (2.1%) and Taeniid eggs (1.05%). Conclusion: A high prevalence rate of parasitic contaminations of herbs in Shushtar necessitates proper washing of herbs and vegetables by consumers to prevent parasitic infections.

Evaluation of Multiplicity of Infection (MOI) and Harvesting Time on the Production of CVS-11 Strain of Rabies Virus in BSR Cell Line

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Rabies is a zoonotic fatal viral disease caused by the rabies virus of the genus Ly... more Introduction: Rabies is a zoonotic fatal viral disease caused by the rabies virus of the genus Lyssavirus, and the family Rhabdoviridae. Challenge virus standard (CVS-11) strain of rabies virus is a key element in rabies reference laboratories, as many gold-standard tests depend on a suitable titer of this strain for interpretation of results. The present study investigated the optimal CVS-11 production in BSR cells (a clone of BHK-21). Methods: We analyzed the kinetic growth of BSR cells in a T-flask and inoculated BSR cells with different MOI of CVS-11 strain of rabies virus, and harvested the produced virus at different time points. Results: Our data showed that BSR cells had a doubling time of around 24-30 h, and at least 95% of cells kept their viability three days after culture. The virus reached the highest titer when the cells were infected at an MOI of 0.1 in DMEM medium, equal to 1.5 × 107 fluorescent focus units (FFU)/ml. Time-course analysis of CVS-11 titer showed that the highest titer was achieved around 72 h post-infection. All tests were performed in triplicate. Conclusion: Since producing the virus in mammalian cell culture is an expensive and complicated method, optimizing the virus production process may be an excellent strategy to lower the cost, save the laboratory resources and maximize productivity.

Comparison of HBV-DNA Levels with Biochemical and Microbiological Parameters for Chronic Hepatitis Evaluation, Bursa, Turkey

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: HBV-DNA levels are used to diagnose chronic hepatitis B (CHB) disease, determine th... more Introduction: HBV-DNA levels are used to diagnose chronic hepatitis B (CHB) disease, determine the infection phase, decide on the treatment, and determine the disease course. We aimed to compare the microbiological and biochemical parameters of patients followed up with chronic hepatitis B pre-diagnosis in our hospital according to their HBV-DNA levels. Methods: HBV-DNA levels were analyzed with the Real-Time PCR method in blood samples of 500 pre-diagnosed CHB patients between February-June 2018, retrospectively. The biochemical parameters of the patients were measured by an automatic biochemical immunoassay analyzer (Beckman Coulter DXI 800, USA), and the microbiological parameters of patients were determined by an automatic analyzer (Roche Cobas 6000, Germany). The differences between the values of biochemical and microbiological parameters of patients were determined according to HBV-DNA. Results: Mean corpuscular volume (MCV) was higher in patients with hepatitis B virus-DNA (HBV-DNA)>20000 IU/mL than patients with negative HBV-DNA, HBV-DNA<2000 IU/mL, and HBV-DNA 2000-20000 IU/mL (P<0.05, P<0.01, P<0.01). Gamma-glutamyl transpeptidase (GGT) was lower in patients with HBV-DNA ranging from 2000-20000 IU/mL than HBV-DNA negative patients, HBV-DNA<2000 IU/mL and HBV-DNA>20000 IU/mL (P<0.05). Albumin was lower in patients with HBV-DNA>20000 IU/mL than patients with HBV-DNA 2000-20000 IU/mL and HBV-DNA<2000 IU/mL (P<0.01). Hepatitis B surface antigen (HBsAg) levels were higher in patients with HBV-DNA 2000-20000 IU/mL than HBV-DNA negative patients, and patients with HBV-DNA<2000 IU/mL and HBV-DNA>20000 IU/mL (P<0.05, P<0.05, P<0.05). Albumin was individually correlated with the HBV-DNA by 2.9%, negatively (P<0.01). Conclusion: MCV, GGT, albumin, HBsAg levels might be used as indicators to diagnose CHB disease, establish the infection phase, decide the treatment, and determine the course of the disease together with HBV-DNA levels.

The Effect of Silibinin on the Expression of TLR7, ISG15, and SOCS1 in Peripheral Blood Mononuclear Cells of Hepatitis C Infected Patients in Comparison with Interferon-α

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Silibinin (silibinin A) is the most active silymarin component, which acts both as ... more Introduction: Silibinin (silibinin A) is the most active silymarin component, which acts both as a hepatoprotective [1] and an antiviral agent. The present study investigated the silibinin effect on IFN-related innate immune genes in PBMCs from HCV-infected patients. Method: 22 chronic HCV patients, including 10 IFN responders and 12 non-responders, were included. Their isolated PBMCs were treated for 6 hours in the presence of silibinin, IFN-α, or their combination. The transcription level of TLR7, ISG15, and SOCS1 genes was compared using real-time PCR. Result: Our result showed that IFN-α induced a significant up-regulation of TLR7 and ISG15 in PBMCs of both responder and non-responder groups. Nevertheless, the SOCS1 gene was not significantly changed in the non-responder group (P=0.32). The combination of IFNα- and silibinin showed a similar pattern to IFN-α alone. By itself, silibinin did not leave a significant change on the expression level of the studied genes. Conclusion: The results indicated that silibinin did not enhance or suppress the expression level of TLR7, ISG15, and SOCS1 genes. Therefore, it has been suggested that its anti-inflammatory role might be devoid of IFN pathways.

Comparison of Antioxidant Activity of Dracocephalum polychaetum Bornm and Nepeta cataria L. and Their Effect on Probiotic Bacteria in a Simulated Gastrointestinal Environment

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Dracocephalum polychaetum Bornm and Nepeta cataria L. are two plants from the Lamia... more Introduction: Dracocephalum polychaetum Bornm and Nepeta cataria L. are two plants from the Lamiaceae family with antibacterial, antifungal, and antiviral properties. This study evaluated the phenolic compounds, antioxidant activity, and effect of aqueous extracts on the survival of Lactobacillus acidophilus and Bifidobacterium animalis in a simulated gastrointestinal environment. Method: The aerial parts of plants were collected at the vegetative growth stage from the Hanza-Kuh's highlands in the Bahr Asman region of Jiroft city, Iran, in spring 2018. The total phenolic content of plants and antioxidant activity were measured using Folin–Ciocalteau and DPPH (2, 2-diphenyl-1-picrylhydrazyl) methods, respectively. For investigating the survival of probiotic bacteria in a simulated gastrointestinal environment, bacterial suspension was inserted into tubes containing 0, 100, 250, 500, and 1000 ppm of extracts and then incubated in a simulated gastrointestinal environment. The probiotic bacteria were counted using an MRS agar medium at various incubation times. Results: The results showed that the amount of total phenolic compounds in the D. polychaetum Bornm extract (44.55 mg/g dry matter) was higher than that of N. cataria L. (18.37 mg/g dry matter). With increased extracts concentrations, the percentage of DPPH-free radicals increased, and D. polychaetum Bornm extract in all concentrations showed higher DPPH free radical inhibitory content compared to the N. cataria L. extract. The viability results in the same gastrointestinal environment showed that samples containing N. cataria extract had a more remarkable survival rate than the controls and D. polychaetum Bornm extract. Conclusion: Using less than 500 ppm of D. polychaetum Bornm and N. cataria L. aqueous extracts can increase probiotic bacteria growth and viability.

Transmission of COVID-19 between Animals and Humans: A challenge for the Scientists

Journal of Medical Microbiology and Infectious Diseases, 2021

In recent decades, some 30 new human pathogens have been identified, of which 75% were spillovers... more In recent decades, some 30 new human pathogens have been identified, of which 75% were spillovers from animals. In late 2019, human infections with a new coronavirus from an unknown origin emerged in China and later spread worldwide. The zoonotic source of severe acute respiratory syndrome coronavirus 2 remains unknown, and there is only some limited information about the close association between the first human cases of COVID-19 and visiting animal markets. Now, bats and pangolins are suspected as natural hosts, and large cats, raccoon dogs, dogs, minks, ferrets, and pangolins as intermediate hosts. There is not enough evidence to prove that animals can transmit COVID-19 infection to humans, but there are some data about the transmission of SARS-CoV-2 between humans and some animal species.

Effect of Host Species on Hatchability of Fasciola hepatica and Fasciola gigantica Eggs from Sheep and Cattle

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Successful development of free-living stages of parasitic helminths depends on larv... more Introduction: Successful development of free-living stages of parasitic helminths depends on larva ability to survive, develop, and hatch. In this study, we aimed to study the host role in the hatching process of Fasciola species. Methods: Fasciola hepatica and Fasciola gigantica eggs were collected from adult worms that originated from naturally infected sheep and cattle livers and were incubated at 26±1°C for 15 days. The percentage of hatched and developed eggs were obtained for each isolate under a light microscope. A polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) was applied to identify the F. hepatica and F. gigantica species. Results: Our findings showed no significant differences in the development rates of F. gigantica and F. hepatica eggs in sheep (69.32% and 72.71%) and cattle (73.56% and74.69%). However, the rates of hatched eggs of F. gigantica and F. hepatica originated from cattle (69.19% and 62.36%) were almost twice the rates in sheep (31.69% and 32.59%), indicating a significant difference. Conclusion: This study demonstrated that host species significantly affect the hatching of Fasciola eggs as the hatching rates of F. gigantica and F. hepatica originated from cattle were higher than those taken from sheep did not affect their larval development. Thus, in addition to environmental factors, the hatching phenomenon is influenced by host species.

Phenotypic Detection of Beta-lactamases among Proteus mirabilis, Enterobacter cloacae, and Citrobacter freundii Isolates from Urinary Samples in Gorgan, Northeast Iran

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: The data on members of the genera Proteus, Pseudomonas, Enterobacter, Citrobacter, ... more Introduction: The data on members of the genera Proteus, Pseudomonas, Enterobacter, Citrobacter, and Staphylococcus as the etiologic agents of urinary tract infections (UTIs) is not much. This study investigated the frequency of various beta-lactamases in urine isolates of Proteus mirabilis, Enterobacter cloacae, and Citrobacter freundii in Gorgan, Golestan province. Methods: A total of 632 urine samples were collected from hospitalized patients in a teaching hospital. The samples were cultured on blood agar and Eosin Methylene blue agar and incubated overnight at 37°C. The cultures with a ≥105 CFU/mL bacterial count were defined as positive for UTI. Bacteria identification was performed using standard biochemical methods and the API20E enteric identification system. The antibiotic resistance pattern was determined by the Kirby-Bauer disk diffusion method, and a phenotypic confirmatory test was used for detecting ESBL, MBL, and AmpC beta-lactamases producers. Results: Out of 632 samples, 317 (50.1%) were positive for UTIs, and 27 (8.5%), 21 (6.6%), and 12 (3.7%) were positive for Enterobacter cloacae, Citrobacter freundii, and Proteus mirabilis isolates, respectively. All the isolates were sensitive to piperacillin-tazobactam and colistin. The prevalence of ESBL and AmpC beta-lactamases in P. mirabilis isolates was higher than the other isolates, but No MBL producers were detected. Conclusions: In this study, the high frequency of ESBL and AmpC beta-lactamases in P. mirabilis isolates may suggest an increasing trend in resistance to cephalosporins and monobactams, which could have a significant impact on the management and treatment of UTI caused by this organism. Therefore, continuous monitoring is required to control the spread of β-lactamase-producing isolates in different geographical areas.

A Fast and Straightforward Method for the Purification of Anti- Immunoglobulin G (IgG) for Coombs Wright Assay

by JoMMID Journal and Rahman Shokri

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Immunoglobulin G is the most abundant immunoglobulin in human blood, comprising app... more Introduction: Immunoglobulin G is the most abundant immunoglobulin in human blood, comprising approximately 75% of serum antibodies. Human IgG is a glycoprotein and can be an antigen for other mammals. Antibody-sensitized red blood cells (RBCs) and complement components can react with anti-human globulins resulting in their agglutination. This study aimed to prepare an antiserum against human IgG for deploying in Coombs Wright and cross-match for rare blood groups. Methods: After isolation of serum from healthy blood donors, serum proteins were precipitated using ammonium sulfate. Consequently, tangential flow filtration and ion-exchange chromatography were applied to purify IgG. SDS-PAGE and Bradford protein content assay was conducted to evaluate the quality and the concentration of the purified IgG. Rabbits were weekly injected with different amounts of the protein four times. Then, sera were obtained from the immunized mice, and total IgG was purified. Finally, the Coombs Wright test was performed on samples from brucellosis patients to validate purified IgG antibody quality. Results: Electrophoresis and Bradford assay results showed that the purified protein had considerable high purity and quantity. Protein bands of reducing and the non-reducing SDS-PAGE showed high purity of the protein along with a protein yield of 2.2 mg/L. Coombs Wright tests using the rabbit anti-human serum had a comparable result with available commercial anti-human immunoglobulin. Conclusion: The results indicated that our method for the purification of IgG was suitable for anti-human globulin preparation. This antibody can also be used in clinical diagnostic tests such as Coombs Wright, cross-match, and blood types evaluation with weak Rh or Du antigens.

Investigation of Chemical Composition, Antibacterial and Antioxidant Activity of Thymus daenensis and Thymus eriocalyx Essential Oils against Human Pathogenic Bacteria

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Plant essential oils can be used as alternative agents for the treatment of antibio... more Introduction: Plant essential oils can be used as alternative agents for the treatment of antibiotic-resistant pathogenic bacteria. This study aimed to investigate the chemical composition, antibacterial, and antioxidant activity of Thymus eriocalyx and Thymus daenensis essential oils against Gram-positive and Gram-negative human pathogenic bacteria. Methods: The aerial part of Thymus eriocalyx and Thymus daenensis in the full flowering stage were collected from West Azerbaijan Province (Urmia), Iran, in 2014. We obtained the essential oils using a Clevenger device. The disc diffusion method was used to determine the antibacterial activity of the essential oils against 15 Gram-positive and Gram-negative PTCC and ATCC bacterial standards. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were measured by microdilution broth method in 96-well plate and free radical scavenging activity by 2,2-diphenyl-1-picrylhydrazyl. Chemical analysis of the essential oils performed out by gas chromatography connected to Mass spectrometry. Results: Eleven (94.06%) and seven (90.76%) compounds were identified in T. eriocalyx and T. daenensis essential oils, respectively. The major components of T. eriocalyx essential oil were thymol (37.8%) and α-terpineol (14.91%), and of T. daenensis were thymol (52.36%) and carvacrol (16.72%). T. eriocalyx essential oil showed the highest activity against B. cereus with MIC of 0.93 μg mL- and MBC of 1.87 μg mL-. The most potent radical scavenging activity was also obtained for T. daenensis essential oil. Conclusion: Essential oil components of T. eriocalyx and T. daenensis may have the potential to be used as antimicrobial agents against antibiotic-resistant pathogenic bacteria.

Ophidascaris sp. in an African Rock Python (Python sebae) in Ghana: A Case Report

by JoMMID Journal and L. Bimi

Journal of Medical Microbiology and Infectious Diseases, 2021

This study reports an Ophidascaris sp. infection in the gastrointestinal tract of a python snake ... more This study reports an Ophidascaris sp. infection in the gastrointestinal tract of a python snake found at a construction site in Labone, a suburb of Accra and the capital city of Ghana. Examination of the rectal contents of the snake by the zinc sulfate centrifugal floatation method revealed helminths eggs. Besides, examining various sections of the alimentary canal revealed adult worms of the genus Ophidascaris in the anterior half of the small intestine just below the stomach's pyloric sphincter. Also, milking one of the female worms onto a microscope slide and examining the exudate by microscopy following staining with Lugol's iodine dye revealed eggs. Our findings may alert the possibility of zoonotic transmission of the parasite from pythons to humans, especially in the suburban/rural areas where people consume snake meat.

Frequency of cbrA, cbrB, ndvB, and phoBR Genes in Relation to Biofilm Formation in Pseudomonas aeruginosa Clinical Isolates

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: After Staphylococcus aureus and Escherichia coli, Pseudomonas aeruginosa is the thi... more Introduction: After Staphylococcus aureus and Escherichia coli, Pseudomonas aeruginosa is the third cause of hospital-acquired infection (HAI). This bacteria's ability to colonize in different environments, especially in hospitals and biofilm formation, has added to its impact as an HAI. The molecular mechanism of biofilm formation is not well understood, but several genes contribute to this phenomenon. This study investigates the frequency of cbrA, cbrB, phoBR, and ndvB genes in biofilm-forming P. aeruginosa isolates. Methods: Fifty P. aeruginosa clinical isolates were collected from various sources such as urine, ulcer, blood, secretions, and trachea in Milad Hospital, Tehran, from 2017 to 2018. Biofilm formation in the isolates was assessed by the microtiter plate assay, and the frequency of cbrA, cbrB, phoBR, and ndvB genes was investigated by PCR. Results: Among the 50 isolates, 44% were strong biofilm former, 34% moderate biofilm former, 12% weak biofilm former, and 10% did not form biofilms. PCR revealed a frequency of 94% for the cbrA gene, 78% for cbrB, 96% for ndvB, and 48% for phoBR. The coexistence of all four genes was 68% in strong biofilm former isolates, 41% in moderate biofilm former isolates, 37% in weak biofilm former, and zero in the isolates that formed no biofilm. Conclusion: The high frequency of ndvB and cbrA genes and the coexistence of ndvB and cbrB suggest the contribution of these genes in the biofilm formation of P. aeruginosa.

In-silico Immunomodelling of SARS-CoV-2

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense si... more Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense single-strand RNA virus belonging to the Coronaviridae family, responsible for coronavirus infectious disease 2019 (COVID-19) with the rapid transmission. This study aimed to characterize and compare SARS-CoV-2 and SARS-CoV major viral proteins and predict antigen proteasomal cleavage patterns, MHC class I processing and presentation, and B T-cell and anti-inflammatory epitopes. Methods: The amino acid sequences of spike surface (S) glycoprotein, membrane (M) glycoprotein, envelop (E) protein, and nucleocapsid (N) phosphoprotein was obtained from NCBI. The sequences were aligned by MEGA 7.0 and modeled by SWISS-MODEL. The proteasomal cleavage pattern, MHC class I processing, and T-cell epitopes were predicted via IEDB analysis and EPISOFT. The B-cell epitopes were predicted by BepiPred 2.0. Also, the prediction of anti-inflammatory epitopes was performed by AntiInflam. Results: Two major antigen proteins, S glycoprotein and M glycoprotein of SARS-CoV-2, respectively, showed 26.57% and 20.59% less efficiency in proteasomal cleavage and presentation to MHC class I, comparing SARS-CoV. There were fewer B-cell predicted epitopes in SARS-CoV-2, comparing SARS-CoV. The anti-inflammatory properties of SARS-CoV-2 S glycoprotein and N protein were higher than SARS-CoV. Conclusion: It seems that the evolution of SARS-CoV-2 is on the way to reducing antigen-presenting to MHC class I and escaping cellular immunity. Moreover, the predicted hotspot epitopes potentially can be used to induce adaptive cellular immunity against SARS-CoV-2. Besides, SARS-CoV-2 appears to be less immunopathogenic than SARS-CoV due to its higher anti-inflammatory proteins.

Comparison of PCR with Serology for Detecting Acute Hepatitis A Virus Infection

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Early detection of acute Hepatitis A virus infection (HAV) allows adopting proper t... more Introduction: Early detection of acute Hepatitis A virus infection (HAV) allows adopting proper treatment measures, rapid recovery, and avoiding side effects. This study compares PCR assay with serology for diagnosing acute HAV infection. Methods: Twenty samples from patients presenting clinical symptoms of acute hepatitis were tested for anti-HAV IgM antibodies. Genomic RNA was extracted from IgM-positive samples, cDNA was synthesized and examined for genomic HAV using a specific HAV real-time detection kit and a nested PCR. Results: Among 20 sera, 14 were positive for anti-HAV IgM antibodies. The specific real-time PCR and nested PCR showed agreement, and both detected HAV genetic material in 3 out of 14 samples. Conclusion: High levels of anti-HAV IgM antibodies do not necessarily indicate acute HAV infection in people presenting clinical symptoms of the disease. Measuring IgM antibody levels alongside molecular detection of virus genome by DNA-based methods assay can lead to an accurate, timely, and reliable diagnosis of active HAV infection.

The Effects of Dehydrozingerone on Growth, Biofilm Formation, and Ergosterol Biosynthesis of Candida albicans

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Candida albicans can cause various diseases, which might lead to various cases of l... more Introduction: Candida albicans can cause various diseases, which might lead to various cases of life-threatening diseases. Biofilm is a specific feature of C. albicans formed on mucosal surfaces and medical devices. Moreover, biofilm protects Candida cells from antifungals and makes the treatment challenging. Here, we studied the effects of dehydrozingerone on C. albicans growth, ergosterol biosynthesis, biofilm formation, and the expression of an essential gene involved in yeast-hypha transition. Methods: C. albicans cells were treated with serial two-fold concentrations of dehydrozingerone (0.125-2 mg/ml) for 48 h at 35 °C. The weights of the fungal cells were estimated as a sign of fungal growth. Biofilm formation was evaluated by a tetrazolium salt (XTT) reduction assay. The expression of the HWP1 gene was assayed by real-time PCR. Results: Dehydrozingerone inhibited C. albicans growth in the range of 3.57% to 84.28%, dose-dependently. The ergosterol content of yeast cells was reduced by 50% in the highest concentration. The biofilm formation was also inhibited by more than 50% at the highest concentration. The expression of the HWP1 gene was suppressed by dehydrozingerone at different concentrations. Conclusion: Our results indicate that dehydrozingerone displayed effective activity against growth, biofilm formation, and ergosterol biosynthesis in C. albicans in vitro.

Prevalence and Antibiotic Susceptibility Profile of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates in Diabetes Patients with Foot Ulcers

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: methicillin-resistant Staphylococcus aureus (MRSA) infection is common among diabet... more Introduction: methicillin-resistant Staphylococcus aureus (MRSA) infection is common among diabetes patients with foot ulcers. This study aimed to determine MRSA isolates prevalence and antibiotic susceptibility profile in diabetic foot ulcers (DFU) patients. Methods: A total of 204 patients with diabetic foot ulcers admitted to a tertiary hospital in Abia State, Nigeria, were included in the assay. Specimens were obtained by scraping the ulcer base or the deep portion of the wound edge using a sterile curette and were promptly sent to the laboratory for culture, identification, and antibiotic susceptibility test. Results: The MRSA prevalence in DFU patients was 22.1% (n=45). Male patients with DFU were more infected with MRSA (n= 26, 12.7%) than females (n=19, 9.3 %), but the difference was not statistically significant (P < 0.14). The age group 41-60 years had the highest prevalence (n=27, 13.2%), statistically significant (P < 0.02). Farmers had the highest prevalence of 9.8% (n=20) while the least (0.5%) was seen in housewives (n=1) with no statistical significance (P < 0.07). The antibiotics sensitivity pattern of MRSA showed 100% sensitivity to vancomycin and chloramphenicol but 100% resistance to penicillin, ceftriaxone, and oxacillin. The multidrug-resistant index was all > 0.2. Conclusions: The prevalence of MRSA in DFU patients in a tertiary hospital in Abia State was very high, with an alarming rate of drug-resistant bacteria due to the possibility of misuse and abuse of antibiotics among the populace, which requires collaborations from all stakeholders to prevent drug resistance in the study settings.

Expression of a Novel HIV-1 Gag-Pol-Env-Nef-Rev Multi-Epitope Construct in Escherichia coli

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Recombinant subunit vaccines have been explored against various human pathogens, ho... more Introduction: Recombinant subunit vaccines have been explored against various human pathogens, however, developing an effective therapeutic toward human immunodeficiency virus (HIV) infection has been challenging. So far, several recombinant HIV-1 antigens have been produced and examined for activation of desired immune responses. This study aimed to express an HIV-1 multiepitope protein as an antigen candidate to develop a vaccine. Methods: In this study, the codon-optimized encoding sequence of the designed multi-epitope construct (Gag-Pol-Env-Nef-Rev) was synthesized and subcloned into the pET-24a (+) expression vector. Then, expression of the target antigen was evaluated in E. coli BL21 (DE3) and Rosetta strains under different conditions (temperature, optical density/ OD600, isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration, and time). Finally, the expression of the Gag-Pol-Env-Nef-Rev multi-epitope protein was confirmed using SDS-PAGE and western blot analysis. Results: The highly conserved and immunodominant T-cell epitopes of HIV-1 Gag, Pol, Env, Nef, and Rev proteins were used to prepare a novel Gag-Pol-Env-Nef-Rev multi-epitope construct. The gag-pol-env-nef-rev gene was successfully sub-cloned in pET-24a (+) vector and subsequently expressed in BL21 (DE3) E. coli strain under optimized conditions (1 mM IPTG, 16 h post-induction, OD 600= 0.6, and 37ºC). A clear band of ~ 35 kDa was detected by western blotting using an anti-His antibody, indicating the successful expression of our target multi-epitope protein. Conclusion: Expression of the recombinant HIV-1 multi-epitope protein was optimized in a bacterial system. The expressed protein will be purified to use as a multi-epitope protein vaccine candidate in the future.

Ghana is Free from the Guinea Worm after a 33-Year Eradication Program

Journal of Medical Microbiology and Infectious Diseases, 2021

For several generations, people from some parts of Ghana have suffered in the hands of a yard-lon... more For several generations, people from some parts of Ghana have suffered in the hands of a yard-long "spaghetti-thin" worm, known as Dracunculus medinensis, which infects humans and leads to a disease known as Dracunculiasis, literally meaning "afflictions with little dragons." The disease, also commonly known as Guinea Worm Disease, Dracuntiasis, or Dracunculosis, is a 3000-year-old known parasitic infection that rarely made headlines before the mid-1980s. Guinea Worm Disease, a plague so ancient that it is believed to be the "fiery serpent torturing the Israelites in the desert," as described in the Old Testament. This paper reviews local and global efforts and strategies at eradicating the disease in Ghana and further diagnoses the problems that hindered the early realization of the desired results of these strategies. This article did not evaluate Ghana's performance in the program. It is equally arduous to unearth all the reasons contributing to the somewhat uneasy road to eradication over three decades of efforts. This review analyzes time-trends, program documents, technical and non-technical reports, and research documents that reveal that Ghana's program ended a decade of disappointing stagnation following the disruptive ethnic conflicts in the early 1990s in its most disease-endemic areas. Despite substantial reductions in the number of guinea worm cases during the mid-1980s, efforts to break the transmission chain in Ghana remained a daunting task. The efforts required continued international and political commitment, active surveillance, strengthening of interventions, and honesty of documenters at all levels.

Transient Gene Expression: an Approach for Recombinant Vaccine Production

Journal of Medical Microbiology and Infectious Diseases, 2021

The production of recombinant vaccines in green plants is an attractive and promising topic in ge... more The production of recombinant vaccines in green plants is an attractive and promising topic in genetic engineering. However, the stable transformation of green plants is a time-consuming, costly, and labor-intensive practice. Moreover, public concerns about genetically modified plants put another limitation on the development and release of transgenic plant-based recombinant vaccines. These shortcomings were addressed by developing transient gene expression systems that allow researchers to investigate candidate recombinant vaccines quickly without tedious work and high costs. A comprehensive literature review was used to gather relevant information. This approach has received much attention in various recombinant vaccine production platforms, including mammalian cell culture, insect cell culture, yeast expression systems, and, more importantly, in plant hosts. Due to their simplicity and efficiency, transient gene expression systems are now widely used to validate gene constructs and transgene expression within plant tissues. This paper describes the concept of transient gene expression and discusses the significant advantages of this approach for producing recombinant vaccines. Notably, the major types of transient gene expression viz. agroinfiltration, viral-based systems, and application of naked plasmid in plant cell culture are introduced, and some examples illustrate the pros and cons of each system. Our literature review also discusses some practical notes on the successful application of this system to provide a more comprehensive image of transient gene expression applicability in green plants. As a whole, this review contributes to the existing literature by shedding more light on various aspects of transient gene expression that have not been addressed thoroughly yet.

Protection and Immune Responses Elicited by rSAG1-PLGA Nanoparticles in C57BL/6 Against Toxoplasma gondii

by JoMMID Journal and Mojgan Allahyari

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: This study aimed to evaluate rSAG1-PLGA efficacy as a particulate vaccine in confer... more Introduction: This study aimed to evaluate rSAG1-PLGA efficacy as a particulate vaccine in conferring protection against Toxoplasma gondii infection in C57BL/6 mice. In light of our previous studies, we studied mice genotype role in eliciting immune responses by rSAG1-PLGA nanoparticles in this study. Methods: Poly (DL-lactide-co-glycolide) (PLGA) nanoparticles loaded by rSAG1 as a subunit vaccine were prepared, and C57BL/6 mice were subcutaneously immunized twice at a 3-week interval by rSAG1-PLGA, soluble rSAG1, blank PLGA, and one group kept unvaccinated. The characteristics of PLGA nanoparticles, the amounts of produced IFN-γ, IL-10, specific anti-ToxoplasmaIgGs, and the conferred protection against infection by T. gondii RH tachyzoite were assessed. Results: rSAG1-PLGA nanoparticles shared a z-average of about 450nm with negative Zeta potential. Compared with the negative control group, the mice vaccinated with rSAG1-PLGA nanoparticles produced significantly higher amounts of IFN-γ, specific anti-T. gondii IgG antibodies and higher titer of IgG2a, which resulted in longer survival times. Conclusion: The efficiency of rSAG1-PLGA nanoparticles in inducing humoral and cellular responses and consequently partial protection against acute toxoplasmosis in C57BL/6 was confirmed.

Contamination of Raw Herbs with Parasitic Protozoa and Helminths in Shushtar City, Southwestern Iran

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Intestinal parasites are among the most prevalent foodborne diseases worldwide, and... more Introduction: Intestinal parasites are among the most prevalent foodborne diseases worldwide, and raw vegetables and herbs are among the primary sources of human infection by these parasites. This study aimed to investigate the prevalence of parasitic contamination of fresh herbs in Shushtar, Khuzestan Province, Southwest of Iran. Methods: In this study, 129 herb samples from various farms were collected and washed with water. The washing waters were centrifuged, and the resulting sediments were examined by formol-ether concentration and Sheather's sugar flotation procedure, as well as a wet smear and Ziehl-Neelsen staining. Results: Among the 129 samples, 73.6% (n=95) showed contamination with at least one parasite, including trophozoite like amoebae (52.6%), followed by Giardia lamblia (14.7%), Cryptosporidium spp. (2.1%), Blastocystis sp. (21%), free-living nematodes larvae (3.1%), Trichostrongilid nematodes (1.05%), Ascaris lumericoids eggs (2.1%), Hymenolepis spp. (2.1%) and Taeniid eggs (1.05%). Conclusion: A high prevalence rate of parasitic contaminations of herbs in Shushtar necessitates proper washing of herbs and vegetables by consumers to prevent parasitic infections.

Evaluation of Multiplicity of Infection (MOI) and Harvesting Time on the Production of CVS-11 Strain of Rabies Virus in BSR Cell Line

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Rabies is a zoonotic fatal viral disease caused by the rabies virus of the genus Ly... more Introduction: Rabies is a zoonotic fatal viral disease caused by the rabies virus of the genus Lyssavirus, and the family Rhabdoviridae. Challenge virus standard (CVS-11) strain of rabies virus is a key element in rabies reference laboratories, as many gold-standard tests depend on a suitable titer of this strain for interpretation of results. The present study investigated the optimal CVS-11 production in BSR cells (a clone of BHK-21). Methods: We analyzed the kinetic growth of BSR cells in a T-flask and inoculated BSR cells with different MOI of CVS-11 strain of rabies virus, and harvested the produced virus at different time points. Results: Our data showed that BSR cells had a doubling time of around 24-30 h, and at least 95% of cells kept their viability three days after culture. The virus reached the highest titer when the cells were infected at an MOI of 0.1 in DMEM medium, equal to 1.5 × 107 fluorescent focus units (FFU)/ml. Time-course analysis of CVS-11 titer showed that the highest titer was achieved around 72 h post-infection. All tests were performed in triplicate. Conclusion: Since producing the virus in mammalian cell culture is an expensive and complicated method, optimizing the virus production process may be an excellent strategy to lower the cost, save the laboratory resources and maximize productivity.

Comparison of HBV-DNA Levels with Biochemical and Microbiological Parameters for Chronic Hepatitis Evaluation, Bursa, Turkey

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: HBV-DNA levels are used to diagnose chronic hepatitis B (CHB) disease, determine th... more Introduction: HBV-DNA levels are used to diagnose chronic hepatitis B (CHB) disease, determine the infection phase, decide on the treatment, and determine the disease course. We aimed to compare the microbiological and biochemical parameters of patients followed up with chronic hepatitis B pre-diagnosis in our hospital according to their HBV-DNA levels. Methods: HBV-DNA levels were analyzed with the Real-Time PCR method in blood samples of 500 pre-diagnosed CHB patients between February-June 2018, retrospectively. The biochemical parameters of the patients were measured by an automatic biochemical immunoassay analyzer (Beckman Coulter DXI 800, USA), and the microbiological parameters of patients were determined by an automatic analyzer (Roche Cobas 6000, Germany). The differences between the values of biochemical and microbiological parameters of patients were determined according to HBV-DNA. Results: Mean corpuscular volume (MCV) was higher in patients with hepatitis B virus-DNA (HBV-DNA)>20000 IU/mL than patients with negative HBV-DNA, HBV-DNA<2000 IU/mL, and HBV-DNA 2000-20000 IU/mL (P<0.05, P<0.01, P<0.01). Gamma-glutamyl transpeptidase (GGT) was lower in patients with HBV-DNA ranging from 2000-20000 IU/mL than HBV-DNA negative patients, HBV-DNA<2000 IU/mL and HBV-DNA>20000 IU/mL (P<0.05). Albumin was lower in patients with HBV-DNA>20000 IU/mL than patients with HBV-DNA 2000-20000 IU/mL and HBV-DNA<2000 IU/mL (P<0.01). Hepatitis B surface antigen (HBsAg) levels were higher in patients with HBV-DNA 2000-20000 IU/mL than HBV-DNA negative patients, and patients with HBV-DNA<2000 IU/mL and HBV-DNA>20000 IU/mL (P<0.05, P<0.05, P<0.05). Albumin was individually correlated with the HBV-DNA by 2.9%, negatively (P<0.01). Conclusion: MCV, GGT, albumin, HBsAg levels might be used as indicators to diagnose CHB disease, establish the infection phase, decide the treatment, and determine the course of the disease together with HBV-DNA levels.

The Effect of Silibinin on the Expression of TLR7, ISG15, and SOCS1 in Peripheral Blood Mononuclear Cells of Hepatitis C Infected Patients in Comparison with Interferon-α

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Silibinin (silibinin A) is the most active silymarin component, which acts both as ... more Introduction: Silibinin (silibinin A) is the most active silymarin component, which acts both as a hepatoprotective [1] and an antiviral agent. The present study investigated the silibinin effect on IFN-related innate immune genes in PBMCs from HCV-infected patients. Method: 22 chronic HCV patients, including 10 IFN responders and 12 non-responders, were included. Their isolated PBMCs were treated for 6 hours in the presence of silibinin, IFN-α, or their combination. The transcription level of TLR7, ISG15, and SOCS1 genes was compared using real-time PCR. Result: Our result showed that IFN-α induced a significant up-regulation of TLR7 and ISG15 in PBMCs of both responder and non-responder groups. Nevertheless, the SOCS1 gene was not significantly changed in the non-responder group (P=0.32). The combination of IFNα- and silibinin showed a similar pattern to IFN-α alone. By itself, silibinin did not leave a significant change on the expression level of the studied genes. Conclusion: The results indicated that silibinin did not enhance or suppress the expression level of TLR7, ISG15, and SOCS1 genes. Therefore, it has been suggested that its anti-inflammatory role might be devoid of IFN pathways.

Comparison of Antioxidant Activity of Dracocephalum polychaetum Bornm and Nepeta cataria L. and Their Effect on Probiotic Bacteria in a Simulated Gastrointestinal Environment

Journal of Medical Microbiology and Infectious Diseases, 2021

Introduction: Dracocephalum polychaetum Bornm and Nepeta cataria L. are two plants from the Lamia... more Introduction: Dracocephalum polychaetum Bornm and Nepeta cataria L. are two plants from the Lamiaceae family with antibacterial, antifungal, and antiviral properties. This study evaluated the phenolic compounds, antioxidant activity, and effect of aqueous extracts on the survival of Lactobacillus acidophilus and Bifidobacterium animalis in a simulated gastrointestinal environment. Method: The aerial parts of plants were collected at the vegetative growth stage from the Hanza-Kuh's highlands in the Bahr Asman region of Jiroft city, Iran, in spring 2018. The total phenolic content of plants and antioxidant activity were measured using Folin–Ciocalteau and DPPH (2, 2-diphenyl-1-picrylhydrazyl) methods, respectively. For investigating the survival of probiotic bacteria in a simulated gastrointestinal environment, bacterial suspension was inserted into tubes containing 0, 100, 250, 500, and 1000 ppm of extracts and then incubated in a simulated gastrointestinal environment. The probiotic bacteria were counted using an MRS agar medium at various incubation times. Results: The results showed that the amount of total phenolic compounds in the D. polychaetum Bornm extract (44.55 mg/g dry matter) was higher than that of N. cataria L. (18.37 mg/g dry matter). With increased extracts concentrations, the percentage of DPPH-free radicals increased, and D. polychaetum Bornm extract in all concentrations showed higher DPPH free radical inhibitory content compared to the N. cataria L. extract. The viability results in the same gastrointestinal environment showed that samples containing N. cataria extract had a more remarkable survival rate than the controls and D. polychaetum Bornm extract. Conclusion: Using less than 500 ppm of D. polychaetum Bornm and N. cataria L. aqueous extracts can increase probiotic bacteria growth and viability.

Transmission of COVID-19 between Animals and Humans: A challenge for the Scientists

Journal of Medical Microbiology and Infectious Diseases, 2021

In recent decades, some 30 new human pathogens have been identified, of which 75% were spillovers... more In recent decades, some 30 new human pathogens have been identified, of which 75% were spillovers from animals. In late 2019, human infections with a new coronavirus from an unknown origin emerged in China and later spread worldwide. The zoonotic source of severe acute respiratory syndrome coronavirus 2 remains unknown, and there is only some limited information about the close association between the first human cases of COVID-19 and visiting animal markets. Now, bats and pangolins are suspected as natural hosts, and large cats, raccoon dogs, dogs, minks, ferrets, and pangolins as intermediate hosts. There is not enough evidence to prove that animals can transmit COVID-19 infection to humans, but there are some data about the transmission of SARS-CoV-2 between humans and some animal species.

Effect of Host Species on Hatchability of Fasciola hepatica and Fasciola gigantica Eggs from Sheep and Cattle

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Successful development of free-living stages of parasitic helminths depends on larv... more Introduction: Successful development of free-living stages of parasitic helminths depends on larva ability to survive, develop, and hatch. In this study, we aimed to study the host role in the hatching process of Fasciola species. Methods: Fasciola hepatica and Fasciola gigantica eggs were collected from adult worms that originated from naturally infected sheep and cattle livers and were incubated at 26±1°C for 15 days. The percentage of hatched and developed eggs were obtained for each isolate under a light microscope. A polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) was applied to identify the F. hepatica and F. gigantica species. Results: Our findings showed no significant differences in the development rates of F. gigantica and F. hepatica eggs in sheep (69.32% and 72.71%) and cattle (73.56% and74.69%). However, the rates of hatched eggs of F. gigantica and F. hepatica originated from cattle (69.19% and 62.36%) were almost twice the rates in sheep (31.69% and 32.59%), indicating a significant difference. Conclusion: This study demonstrated that host species significantly affect the hatching of Fasciola eggs as the hatching rates of F. gigantica and F. hepatica originated from cattle were higher than those taken from sheep did not affect their larval development. Thus, in addition to environmental factors, the hatching phenomenon is influenced by host species.

Phenotypic Detection of Beta-lactamases among Proteus mirabilis, Enterobacter cloacae, and Citrobacter freundii Isolates from Urinary Samples in Gorgan, Northeast Iran

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: The data on members of the genera Proteus, Pseudomonas, Enterobacter, Citrobacter, ... more Introduction: The data on members of the genera Proteus, Pseudomonas, Enterobacter, Citrobacter, and Staphylococcus as the etiologic agents of urinary tract infections (UTIs) is not much. This study investigated the frequency of various beta-lactamases in urine isolates of Proteus mirabilis, Enterobacter cloacae, and Citrobacter freundii in Gorgan, Golestan province. Methods: A total of 632 urine samples were collected from hospitalized patients in a teaching hospital. The samples were cultured on blood agar and Eosin Methylene blue agar and incubated overnight at 37°C. The cultures with a ≥105 CFU/mL bacterial count were defined as positive for UTI. Bacteria identification was performed using standard biochemical methods and the API20E enteric identification system. The antibiotic resistance pattern was determined by the Kirby-Bauer disk diffusion method, and a phenotypic confirmatory test was used for detecting ESBL, MBL, and AmpC beta-lactamases producers. Results: Out of 632 samples, 317 (50.1%) were positive for UTIs, and 27 (8.5%), 21 (6.6%), and 12 (3.7%) were positive for Enterobacter cloacae, Citrobacter freundii, and Proteus mirabilis isolates, respectively. All the isolates were sensitive to piperacillin-tazobactam and colistin. The prevalence of ESBL and AmpC beta-lactamases in P. mirabilis isolates was higher than the other isolates, but No MBL producers were detected. Conclusions: In this study, the high frequency of ESBL and AmpC beta-lactamases in P. mirabilis isolates may suggest an increasing trend in resistance to cephalosporins and monobactams, which could have a significant impact on the management and treatment of UTI caused by this organism. Therefore, continuous monitoring is required to control the spread of β-lactamase-producing isolates in different geographical areas.

A Fast and Straightforward Method for the Purification of Anti- Immunoglobulin G (IgG) for Coombs Wright Assay

by JoMMID Journal and Rahman Shokri

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Immunoglobulin G is the most abundant immunoglobulin in human blood, comprising app... more Introduction: Immunoglobulin G is the most abundant immunoglobulin in human blood, comprising approximately 75% of serum antibodies. Human IgG is a glycoprotein and can be an antigen for other mammals. Antibody-sensitized red blood cells (RBCs) and complement components can react with anti-human globulins resulting in their agglutination. This study aimed to prepare an antiserum against human IgG for deploying in Coombs Wright and cross-match for rare blood groups. Methods: After isolation of serum from healthy blood donors, serum proteins were precipitated using ammonium sulfate. Consequently, tangential flow filtration and ion-exchange chromatography were applied to purify IgG. SDS-PAGE and Bradford protein content assay was conducted to evaluate the quality and the concentration of the purified IgG. Rabbits were weekly injected with different amounts of the protein four times. Then, sera were obtained from the immunized mice, and total IgG was purified. Finally, the Coombs Wright test was performed on samples from brucellosis patients to validate purified IgG antibody quality. Results: Electrophoresis and Bradford assay results showed that the purified protein had considerable high purity and quantity. Protein bands of reducing and the non-reducing SDS-PAGE showed high purity of the protein along with a protein yield of 2.2 mg/L. Coombs Wright tests using the rabbit anti-human serum had a comparable result with available commercial anti-human immunoglobulin. Conclusion: The results indicated that our method for the purification of IgG was suitable for anti-human globulin preparation. This antibody can also be used in clinical diagnostic tests such as Coombs Wright, cross-match, and blood types evaluation with weak Rh or Du antigens.

Investigation of Chemical Composition, Antibacterial and Antioxidant Activity of Thymus daenensis and Thymus eriocalyx Essential Oils against Human Pathogenic Bacteria

Journal of Medical Microbiology and Infectious Diseases, 2020

Introduction: Plant essential oils can be used as alternative agents for the treatment of antibio... more Introduction: Plant essential oils can be used as alternative agents for the treatment of antibiotic-resistant pathogenic bacteria. This study aimed to investigate the chemical composition, antibacterial, and antioxidant activity of Thymus eriocalyx and Thymus daenensis essential oils against Gram-positive and Gram-negative human pathogenic bacteria. Methods: The aerial part of Thymus eriocalyx and Thymus daenensis in the full flowering stage were collected from West Azerbaijan Province (Urmia), Iran, in 2014. We obtained the essential oils using a Clevenger device. The disc diffusion method was used to determine the antibacterial activity of the essential oils against 15 Gram-positive and Gram-negative PTCC and ATCC bacterial standards. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were measured by microdilution broth method in 96-well plate and free radical scavenging activity by 2,2-diphenyl-1-picrylhydrazyl. Chemical analysis of the essential oils performed out by gas chromatography connected to Mass spectrometry. Results: Eleven (94.06%) and seven (90.76%) compounds were identified in T. eriocalyx and T. daenensis essential oils, respectively. The major components of T. eriocalyx essential oil were thymol (37.8%) and α-terpineol (14.91%), and of T. daenensis were thymol (52.36%) and carvacrol (16.72%). T. eriocalyx essential oil showed the highest activity against B. cereus with MIC of 0.93 μg mL- and MBC of 1.87 μg mL-. The most potent radical scavenging activity was also obtained for T. daenensis essential oil. Conclusion: Essential oil components of T. eriocalyx and T. daenensis may have the potential to be used as antimicrobial agents against antibiotic-resistant pathogenic bacteria.