Journal of Medical Bacteriology

Frequency of Shiga Toxin Associated Genes in Escherichia coli Isolated from Salivary Abomasum Disease in Kid Goats

Saba Almasi Chegeni — 2024-11-16

Background: Salivary abomasum disease (SAD) is a devastating disease causing significant mortality in Iranian goat and sheep herds. Understanding the causative agents is essential for developing effective preventive measures. This study investigated the potential role of Shiga toxin-producing Escherichia coli (STEC) in SAD pathogenesis.

Methods: We isolated E. coli from kid goats aged 3-30 days experiencing a sudden, acute illness characterized by gait imbalance, and death within 48 hours during the kidding season. Following isolation, we employed multiplex PCR to identify the presence of Shiga toxin genes (Stx1 and Stx2) associated with virulence in STEC strains.

Results: E. coli was isolated from 30 (75%) out of 40 animals. Notably, 7 (23.3%) isolates harbored the Stx2 gene, while only one isolate (3.3%) possessed the Stx1 gene.

Conclusion: These findings suggest a potential role for STEC, particularly strains carrying the Stx2 gene, in the development of SAD and multiple abomasal hemorrhages, in kid goats. The presence of Shiga toxin genes in a significant proportion of E. coli isolates highlights the importance of further research to elucidate their contribution to SAD pathogenesis and inform the development of targeted interventions.

Diagnosis of Chlamydia abortus by Isolation in Cell Culture and Real Time PCR in Aborted Sheep and Goats

Hossein Esmaeili — 2024-11-16

Background: Ovine enzootic abortion (OEA) caused by Chlamyidia abortus is one of the most important abortive disease in small ruminants. Diagnosis of Ovine enzootic abortion depends on the isolation and detection of the agent or its nucleic acid. The aim of the present study was to detect Chlamydia abortus using both isolation method and real-time PCR in Brucella free flocks in Iran.

Methods: Twenty-eight vaginal and conjunctival swab samples which were Chlamydia abortus seropositive, were selected from ewes and does with recent abortion. Then the samples were tested by real-time PCR and positive molecular samples were inoculated into McCoy cells.

Results: Using real-time PCR, 18 samples (64.3%) were positive and 7 (25%) of them were isolated in cell culture.

Conclusion: The present results indicate that Chlamydia can play a relatively significant role in the abortion in does and ewes in Iran. Although the isolation of Chlamydia abortus have 100% specificity, because of low sensitivity, time consuming, cost and high probability of contamination, it is not suitable for routine laboratory diagnosis. Therefore, applying real-time PCR which have high sensitivity and specificity is recommended.

Impact of Nano-Pomegranate Seed Oil on the Expression of TLR2 and TLR4 Genes in A549 Cells Sensitized with Alternaria alternata Cellular Extract

Seyed Mehdi Joghataei — 2024-11-16

Background: Allergies impact nearly 30% of the world's population, with fungi being remarkable contributors to allergic sensitivity. Exposure to fungi allergens can trigger allergic reactions and severe asthma has been linked to hypersensitivity to fungi such as Aspergillus and Alternaria spp. Alternaria alternata, a common allergen in respiratory allergic diseases, releases proteases that initiate Th2 responses, causing inflammatory cytokine production. Given the anti-inflammatory properties of Pomegranate Seed Oil (PSO) and the potential of Nano-emulsions (NEs) to enhance drug delivery, this study investigates the impact of PSO-loaded NEs on TLR2 and TLR4 gene expression in A549 cells sensitized with A. alternata extract (ALT).

Methods: A. alternata (ATCC 6663) was cultured and processed to obtain a cytosolic extract, with protein content measured using the Bradford method. Using a Soxhlet apparatus, PSO was extracted from cleaned, dried seeds and analyzed by gas chromatography. Alginate nanospheres containing PSO were prepared through a modified water-in-oil emulsification method and characterized for particle size, polydispersity index, and zeta potential. A549 cells were cultured and treated with various ALT, PSO, and PSO-loaded NEs combinations. Following treatment, RNA was extracted, and real-time RT- PCR was conducted to analyze TLR2 and TLR4 gene expression.

Results: All treatment groups showed an increase in TLR2 gene expression compared to the control, with the ALT combined with PSO (P+ALT) causing the highest increase at 4.82-fold. Free PSO (P) and free NEs (NP) resulted in 3.92-fold and 2.93-fold increases, respectively, while the ALT and PSO- loaded NEs (NP+ALT) led to 2.26-fold and 2.50-fold increases. For TLR4 gene expression, the ALT treatment increased expression by 2.29-fold, but treatments containing PSO (P, P+ALT, NP+ALT) reduced TLR4 expression, with P+ALT and NP+ALT causing 0.45-fold and 0.61-fold decreases.

Conclusion: The study confirms that herbal extracts like PSO selectively upregulate TLR2 and downregulate TLR4, suggesting targeted therapeutic potential in inflammation and immune modulation. PSO-loaded NEs demonstrated superior anti-inflammatory effects, supporting their development for treating inflammatory diseases and warranting further research into their molecular mechanisms and therapeutic applications.

Exploring the Phytochemical Profile and Antimicrobial Potential of Leaf Extracts from Megaphrynium macrostachyum

Salisu Danjuma Ibrahim — 2024-11-16

Background: Medicinal plants offer a promising reservoir of bioactive compounds, placing them as a compelling avenue for novel drug exploration. In recent times, the emphasis on harnessing natural products sourced from medicinal plants has escalated due to their diminished adverse effects, economic viability, and efficacy against a broad spectrum of antibiotic-resistant microorganisms. The aim of this investigation was to scrutinize the phytochemical constitution and antimicrobial efficacy inherent to Megaphrynium macrostachyum leaves.

Methods: Three distinct solvents – ethanol, water-ethanol, and water – were employed to extract the diverse range of phytochemicals housed within the leaves. Subsequently, the extracted compounds were subjected to assessment for their antimicrobial potential against both bacteria and fungi, which were isolated from various samples. This evaluation was executed employing the agar well diffusion method.

Results: The qualitative analysis of phytochemical components unveiled the substantial presence of alkaloids, terpenoids, phenols, tannins, flavonoids, while a relatively lower occurrence of steroids was observed across the different leaf extracts. Further quantitative analysis showed that the most potent extract exhibited elevated phenolic content (2.400 mg/ml), closely trailed by flavonoids (1.995 mg/ml) and saponins (1.909 mg/ml). This study furnishes compelling proof of the efficacy encompassed within Megaphrynium macrostachyum leaves, particularly concerning their proficiency against both fungi and bacteria.

Conclusion: As such, it adds momentum to the accumulating body of knowledge concerning the
exploitation of natural medicinal plants, paving the way for enhanced therapeutic interventions.

Comparison of RT.PCR and ELISA Methods in the Diagnosis of Human Cytomegalovirus in Kidney Transplant Patients

Fatemeh Bali Chalandar — 2024-11-16

Background: In kidney transplant recipients prone to infections like cytomegalovirus (CMV), a vital need for an accurate diagnostic method is evident. This study at Khorshid Laboratory in Tehran rigorously compares real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) for CMV detection in transplant patients.

Methods: In January to March 1400, 70 kidney transplant recipients were assessed for CMV DNA using RT-PCR and concurrent ELISA tests, with statistical analysis aided by SPSS.

Results: In kidney transplant patients (average age: 49.40 ± 13.64 years), 4.3% tested positive for CMV via PCR. Strong correlation between serological and molecular methods. IgG and IgM antibody detection both showed high sensitivity and specificity, advocating for efficient CMV diagnosis at Khorshid Laboratory, Tehran.

Conclusion: This study emphasizes the need for a quick and efficient CMV diagnostic approach in kidney transplant patients. The strong correlation between molecular and serological methods favors using the faster RT-PCR method, crucial for timely management, especially with the increasing age- related CMV incidence. The findings strongly recommend RT-PCR integration for enhanced sensitivity in kidney transplant patients at Khorshid Laboratory, Tehran.

Effects of Topical Polymyxin Neomycin Hydrocortison in Patients with Non-Allergic Rhinitis, a Randomized Clinical Trial

Sara Mollazadeh — 2024-11-16

Background: Bacterial infection is involved in the pathogenesis of inflammatory non-allergic rhinitis
(NAR), and a broad-spectrum antibiotic should reduce the severity of the disease by eliminating the
mucosal biofilm. The purpose of this study is to assess the efficacy of a topical mixture of polymyxin,
neomycin, and hydrocortisone in patients with NAR.

Methods: This double-blind phase 3 randomized clinical trial was conducted on 90 patients with non- allergic rhinitis (NAR) in Mashhad, Iran. The subjects experienced symptoms for more than two weeks, and the pretrial course of topical corticosteroids was ineffective. Patients were randomly assigned to treatment groups receiving polymyxin NH, hydrocortisone, or saline. The drug was administered as nasal drops three times daily for a treatment course of two weeks, and the containers were identical. The primary outcome was nasal obstruction.

Results: The pretrial comparison showed no significant difference between groups in terms of clinical findings. However, after the trial, nasal obstruction, as the primary outcome, significantly decreased from 20 subjects (66%) to 7 subjects (23%) in the Polymyxin NH group, along with other secondary outcomes, including palatal itching (56% to 23%), sneezing (76% to 40%), mucosal inflammation (100% to 75%), post-nasal drip (PND) (96% to 63%), and concha swelling (96% to 73%). In the other groups, sneezing was the only significant improvement observed in the saline group. Cytology of nasal discharge showed a reduction in nasal neutrophil counts in the Polymyxin NH group (68±16.7% to 46±27.6%) compared to the hydrocortisone and saline-treated groups.

Conclusion: Topical intranasal polymyxin NH is effective in treating patients with non-allergic rhinitis.

The Prevalence of Fluoroquinolone-Resistant E. coli in Animals and Animal Products: A Systematic Review and Meta-Analysis

Hadi Sedigh Ebrahim-Saraie — 2024-11-16

Background: Biological contamination of foods is a serious problem for human health. Animal and animal products may be contaminated by these biological and chemical contaminants. One of the most important causes of foodborne illness in humans is Escherichia coli. Fluoroquinolones can be used as a suitable treatment for enteric infections in food-producing livestock. We aimed to evaluate the current status of resistance of E. coli strains isolated from animals and animal products to fluoroquinolone in Iran.

Methods: A systematic search was conducted using the Web of Science, PubMed, Embase, Google Scholar, and Scopus databases from 2000 to Oct 2020. Nineteen studies were selected based on the inclusion criteria and analysis by Comprehensive Meta-Analysis.

Results: Based on the data analysis, The rates of antibiotic resistance in animal strains were as follows: Flumequine (75.1%), Enrofloxacin (55.2%), Danofloxacin (48.1%), Ciprofloxacin (48.4%), and Norfloxacin (52.9%). Next, the rates of quinolone resistance among E. coli strains isolated from animal products were Norfloxacin (45.5%), Ciprofloxacin (44.5%), and Enrofloxacin (60.9%). Based on the funnel plots and Egger's test, there was no significant publication bias.

Conclusion: We finally concluded that antibiotic resistance in commensal E. coli is related to the overuse of antibiotics in livestock, especially fluoroquinolones.

Possible Link between Gut Microbiota, Diet, and COVID-19 Infection

Paria Fazli — 2024-11-16

Background: Coronavirus disease 2019 (COVID-19) is a concern for world health since it may impact both the upper (nose, throat, sinuses) and lower (trachea, lungs) respiratory tracts. Death (at a rate of 10%), respiratory failure, multi-organ failure, and acute respiratory distress syndrome (ARDS) are among the problems that might arise. Recent years have seen a global spread of zoonotic coronaviruses, which have caused human epidemics such as MERS and SARS. Various clinical symptoms may be seen in this sickness because to the numerous changes in intestinal homeostasis caused by SARS-CoV-2. Because of the beneficial impact that probiotics have on the host immune system, gastrointestinal disorders may now be effectively treated. This article discusses the close relationship between what we eat, the bacteria in our gut, and the risk of contracting the COV-19 virus.

Conclusion: The relationship between gut microbiota, dietary factors, and COVID-19 severity indicates that the microbiome may influence immune regulation. Imbalances in microbial communities and reduced diversity can intensify inflammation, potentially worsening COVID-19 outcomes. Strategies such as probiotics, prebiotics, and dietary changes might provide therapeutic benefits by improving gut health and strengthening immune defenses. However, further research is required to clarify these mechanisms and establish effective interventions.