ACTA Microbiologica Hellenica (Volume 65, Issue 1)

George Pappas1, Georgia Vrioni2

1Institute of Continuing Medical Education of Ioannina, Greece.

2Department of Microbiology, Medical School, National and Kapodistrian University of Athens, Greece.

Ioannis Daniil, Aggeliki Georgaki, Vaia Papadouli, Nektaria Rekleiti, Vasiliki Mamali,Olympia Zarkotou, Kate Themeli-Digalaki

Microbiology Department of “Tzaneio” General Hospital of Piraeus

Influenza is an acute viral infection of the respiratory system, which is mainly caused by influenza viruses type A and B (ΙΝΑ, ΙΝΒ) in humans, with high transmissibility. Seasonal influenza outbreaks cause significant mortality and morbidity each year. The aim of the present study was to record the incidence of seasonal influenza cases in General Hospital of Piraeus “Tzaneio” and their epidemiological study. From the 14th December 2018 to the 29th April 2019, a total of 194 influenza suspected patients (100 males and 94 females, mean age ± S.D., 42.6 ± 29.2 y) were examined for influenza viruses. Direct nasal or nasopharynx smears were tested for influenza viruses using either immunochromatographic assay (ΙCG-CORIS BioConcept, Belgium) or molecular method (Alere-i, Abbott, US) or both. 78/194 (40.2%) patients were found to bepositive for INA/ΙΝB either by ΙCG assay (28/101 patients, 27.7%) or by Alere-i (50/115 patients, 43.5%). The majority (77/78 patients) were positive for INA, while only one patient was positivefor INB. Moreover, 31/78 (39.7%) influenza cases occurred in the age group of 15-64 y, followedby the age group of ≥ 65 y with 26 (33.3%) cases. The peak of influenza outbreak (18 cases) was observed during the 4th week of 2019 (21 – 27/1/2019). Mortality among patients diagnosed with influenza was 5.1% (4 of 78 infected patients died). Finally, three of 4 deaths were observed in patients hospitalized or need to be treated in ICU. In conclusion, the cases of influenza recorded in our hospital were caused mainly by the ΙΝΑ virus, as observed throughout the country, culminating during the last 10 days of January 2019. The combination of the clinical assessment about severity of illness and the using of appropriate diagnostic method with high sensitivity and specificity, depending on the patient’s profile (such as age, antiviral treatment, severity of the disease, and risk factors) contributes to timely and reliable diagnosisof influenza, in order to its quickly control and optimal treatment of patient.

Keywords: Influenza, Epidemiology, Molecular Diagnostics, Immunoassay

Ioannis Daniil1, Efstathios Laskos1, Stefanos Charpantidis1, Triantafyllia Pitsia1, Maria Balachouti2,Georgia Vrioni3,4, Dikaia Neamonitou1, Michail Dimoutsos1

1Department of Microbiology and Biochemistry, Psychiatric Hospital of Attica “Dromokaition”, Haidari, Greece

2Committee for Healthcare-associated Infections, Psychiatric Hospital of Attica “Dromokaition”, Haidari,

Greece

3Sexually Transmitted Diseases and AIDS Laboratory, Hospital for Skin and Venereal Diseases “Andreas Syngros”,

Athens, Greece

4Department of Microbiology, Medical School, National and Kapodistrian University of Athens, Greece.

Syphilis still remains a major healthconcern worldwide because of the possibility of seriousmedical and psychological consequences. Neurosyphilis can present with psychiatric symptomsalone that can mimic any other mental illness. Furthermore, psychiatric patients are atan increased risk of sexually transmitted diseases (STDs) like syphilis, human immunodeficiencyvirus (HIV) infection, hepatitis B and hepatitis C. Co-infections of these STDs are indeeddangerous causing severe and difficult-to-treat complications. The aim of this study is to investigatethe prevalence and epidemiological characteristics of syphilis and its co-infectionscaused by HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV) among hospitalized patientswith mental illness. During a 4-year period (2015-2018) 7,669 hospitalized patients of PsychiatricHospital of Attica “Dromokaition” in Greece were examined serologically for the presenceof antibody to Treponemapallidum(TP), HIV Ag/Ab, HbsAg and Anti-HCV using the chemiluminescentmicroparticleimmunoassay method (CMIA, Architect i1000SR, Abbott). Serologicconfirmation of syphilisperformed in the Laboratory ofHospitalfor SkinandVenereal Diseases“Andreas Sygros”, using two treponemal tests (ELISA and TPPA) and one non-treponemalmethod (VDRL), while the molecular confirmation of HIV positive samples performed in theNational HIV/AIDS Reference Laboratory of Hellenic National Public Health Organization(EODY/KEELPNO). Statistical analysis was performed using IBM SPSS Statistics v. 22.0. 65/7,669(0.85%) psychiatric patients were confirmed having syphilis and male patients were more frequentlyinfected. 15/65(23.0%) patients with syphilis were coinfectedwith viral STDs (13 patientswith HCV and 2 patients with HIV and none with HBV infections). The mean age ofpatients with co-infections was lower than the patients having a single syphilis (49±11.2y and55±14.1 y, respectively). Intravenous drug users (IDUs) (p<0.001; OR: 43.083 [95% CI=8.403 –220.898] and sex-workers (p=0.044; OR: 5.697 [95% CI=1.111 – 29.210] were significantly relatedto the co-infections. In conclusion, syphilis is not uncommon in our psychiatric population.HCV or HIV co-infection prevalence rates were high among patients with syphilis, usuallyaffected younger patients, IDUs and sex workers. None HBV co-infection occurred in this study,possibly due to the ongoing HBV vaccination program. Keywords: syphilis, co-infection, psychiatric patients,hepatitis, HIV

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Roxana Mansour Ghanaiea, Leila Azimia, SeyedehMahsanHoseini‐Alfatemia, Ahmad MoeinKarimib,GoliAngotic, IrajSedighid, Reza Daliranie, Ahmad Shamsizadehf, AbdollahKarimia

aPediatric Infections Research Center, Research Institute for Children’s Health, ShahidBeheshti Universityof Medical Sciences, Tehran, Iran

bIslamic Azad University, Tehran Medical Science Branch, Tehran, Iran

cKhatamolanbia Hospital, Mianeh, Iran

dDepartment of Pediatric, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.

eMofid Children Hospital, ShahidBeheshti University of Medical Sciences, Tehran, Iran

fInfectious and Tropical Diseases Research Center, Health Research Institute, Jundishapur University of Medical Sciences, Ahvaz, Iran

Brucellosis or Malta fever is a life-threatening systemic human infection and one of the mostcommon zoonotic diseases worldwide, causing considerable public health and economic problems.

The Determination of antibiotic susceptibility profile of Brucellaspp. in each region orcountry is clinically important and has essential role in epidemiological studies and infectioncontrol policy planning. The present study aimed to determine the occurrence of the mostcommon antimicrobial resistance genes among Brucella spp. isolated from Iranian patients.

This cross-sectional multicenter study was conducted on non-repetitive blood specimen frompatients suspected to have brucellosis for a period of two years from January 2015 to January2017 referred to hospitals in Tehran, Ahvaz, Hamadan, Qazvin and Mianeh in Iran. The presenceof mutations in gyrA, parC, rpoB, and aadA1 genes within Brucellaisolates were investigated byPCR method. Ninety-one blood samples were included in this study. Presence of the Brucellaspp. was confirmedin 52 (55%) samples molecularly. PCR screening for the presence of antibiotic resistancemechanisms showed that 26 (50%), 4 (7.7%) and 2 (3.8%) of isolates were positive for aadA1,parC and gyrA genes, respectively. None of the isolates contained rpoB genes. Concurrent presence

ofparC/aadA1genes was found in 3 (5.8%) of isolates.

In conclusion, this is the first study to investigate the frequency of antibiotic resistance mutationsin Brucella isolates in Iran, detecting antibiotic resistance-inducing mutations to commonlyused antibiotics.

Keywords: Brucellosis, Brucella, Antibiotic resistance, Mutation

Abdelhafid Boubendir1, Mohammed Mostakim2

1Laboratory of Natural Sciences and Materials, University Centre AbdelhafidBoussouf, Mila. RP. 26. Mila, 43000, Algeria

2Higher Institute of Nursing Profession and Techniques of Health, Errachidia. BP: 57 or BP: 24. Errachidia,52000, Morocco

Carbapenem antibiotics are considered last resort options in the treatment of infections causedby multidrug-resistant Enterobacteriaceae producing Extended Spectrum β-Lactamases (ESBLs). The emergence of Klebsiellapneumoniae OXA-48 in particular is steadily expanding and represents a major public health concern. The aim of the present study is to analyze the variability

and phylogeny of K. pneumoniae OXA-48 amino acids structures from different geographic areas of the world.

The data on K. pneumoniae OXA-48 amino acids structures were collected during the month of May 2019 from the Protein Data Bank (PDB). The alignment of protein sequences was carried using the Clustal Omega program available on the UniProt database. The phylogenetic analysis and dendrogram were realized using MEGA software version 6.

Among 58 structures, 8 representative OXA-48 variants were selected for the study. The alignment demonstrated that the conserved motifs were in general well conserved except for the two mutations S70G and S70A remarked respectively in the two chains 5HAQ and 5HAP from the United States. However, the OXA-181 and OXA-245 variants displayed mutations far away

from the active sites. In comparison with OXA-48, OXA-181 variant showed four substitutions at Thr104Ala, Asn110Asp, Glu168Gln, and Ser171Ala; while OXA-245 had a single amino acid substitution Glu125Tyr. The phylogenetic analysis revealed three distinct clusters; the first one consists of four OXA-48 structures (Canada, Norway, United States and Italy) and one OXA-245

(Norway), the second includes two OXA-48 structures from the United States, while the thirdcluster is formed by an individual OXA-181 from Norway.

The results of this study confirm a similar evolutionary trend in the structure of K. pneumonia OXA-48 variants worldwide. The current data on K. pneumoniae OXA-48 amino acids structures is limited to restricted geographic areas, and need broadening to provide the actual state of molecular changes and antimicrobial resistance evolution.

Keywords: Klebsiellapneumoniae, OXA-48, antibiotic resistance, phylogeny

Iryasti Yudistia1, Tri YudaniMardining Raras2, Tri Wahju Astuti3

1Master Program in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Malang, Indonesia.

2Department of Biochemistry and Molecular Biology, Faculty of Medicine, Brawijaya University, Malang, Indonesia.

3Department of Pulmonology.

Beneficial bacteria such as Lactobacillus spp. may exhibit promising therapeutic potential due to their inhibitory effects against antibiotic-resistant microorganisms. The present study aimed to evaluate the potency of Lactobacillus helveticus C2 isolated from kefir against multidrug resistant

Klebsiellapneumoniae. The ability of L. helveticus C2 to inhibit MDR K. pneumoniaeadhesion to intestinal enterocytes was observed in vitro. Intestinal colonization by MDR K.pneumoniae in the presence of L. helveticus C2 was further evaluated using a mice model. L.helveticus C2 exhibited strong antibacterial activity against MDR K. pneumoniae. Direct antagonism between L. helveticus C2 and MDR K. pneumoniae cells could not be ascertained in vitro.

However, the adhesion of MDR K. pneumoniae to the small intestinal enterocytes of mice was inhibited by pili and outer membrane protein of L. helveticus C2. Moreover, L. helveticus C2demonstrated inhibitory activity against MDR K. pneumoniae colonization by reducing the number of MDR K. pneumoniae colonies in BALB/c mice as induced by pellets and cell-free supernatant of L. helveticus C2. L. helveticus C2 is a probiotic strain of significant therapeutic or preventive potential due to its strong antibacterial activity, as well as the ability to inhibit the adhesion and colonization of MDR K. pneumoniae.

Keywords: Anti-bacterial, Anti-adhesion, Anti-colonization, MDRKlebsiellapneumoniae, Lactobacillus helveticus C2