Phenotypic and Molecular Characterization of Extended-Spectrum beta-Lactamase Produced by Escherichia coli, and Klebsiella pneumoniae Isolates in an Educational Hospital

Background: Extended-spectrum beta-lactamases (ESBLs) are a group of enzymes that hydrolyze antibiotics, including those containing new cephalosporins, and they are found in a significant percentage of Escherichia coli and Klebsiella pneumoniae strains. With the widespread use of antibiotics, difficulties with infection therapy caused by drug resistant organisms, especially those that have acquired resistance to beta-lactams, such as broad-spectrum cephalosporins, have amplified the above-mentioned organisms. Objectives: This study was conducted to characterize ESBLs among E. coli and K. pneumonia isolates by molecular and phenotypic methods. Materials and Methods: Different strains of E. coli and K. pneumonia were collected from patients with urinary tract infections. The ESBL phenotype was determined by a double disk diffusion test (DDDT). In addition, polymerase chain reaction (PCR) analysis specific for beta-lactamase genes of the TEM and SHV family was carried out. The PCR products were run on agarose and examined for DNA bands. Results: A total of 245 E. coli and 55 K. pneumonia strains were isolated from different samples. In total, 128 of the 300 isolates were confirmed as potential ESBLs producers as follows: 107 (43.67%) E. coli and 21 (38.18%) K. pneumonia. ESBLs genes were found in 24 isolates (18.75%): 21 E. coli and 3 K. pneumonia isolates. The TEM gene was present in 13 (12.14%) E. coli strains, but it was not detected in K. pneumonia. In addition, the SHV gene was present in 8 (7.47%) E. coli and 3 (14.28%) K. pneumonia isolates. Five (4.67%) of the E. coli isolates harbored both TEM and SHV genes. All isolates (100%) were susceptible to imipenem. The lowest rates of resistance to other antibiotics were observed for; piperacillin-tazobactam (6.25%), amikacin (12.5%) and gentamicin (14.84%). The rates of resistance to other antibiotics were as follow: nitrofurantoin (16.4%), nalidixic acid (23.43), co-trimoxazole (25%), cefepime (32%), ciprofloxacin (55.46%), ampicillin (69.53%), ceftazidime (100%), and cefotaxime (100%). Conclusions: The results of this study indicate the widespread prevalence of ESBLs and multiple antibiotic resistance in E. coli and K. pneumoniae. Therefore, beta-lactam antibiotics and beta-lactamase inhibitors or carbapenems should be prescribed based on an antibacterial susceptibility test

The effect of verapamil on in vitro susceptibility of promastigote and amastigote stages of Leishmania tropica to meglumine antimoniate

Pentavalent antimonials are the standard treatment for cutaneous leishmaniasis (CL) with low efficacy and resistance is emerging. CL is increased significantly in respect to incidence rate and expanding to new foci. In the present study, the effect of verapamil on in vitro susceptibility of promastigote and amastigote stages of Leishmania tropica to meglumine antimoniate (MA, Glucantime) was evaluated using colorimetric assay (MTT) and in a macrophage model, respectively. Verapamil, as a calcium channel blocker, affects drug uptake by preventing of drug efflux from the cells. In promastigote form, several concentrations of MA with or without verapamil showed significant decrease (P<0.05) in optical density. The overall mean IC50 value with combination of MA plus verapamil (IC50=116.03 μg/ml) was significantly less than MA (IC50=225.14 μg/ml) alone (P<0.05) for promastigote stage. Similarly, the amastigote stage was more susceptible to treatment with MA plus verapamil to that of MA alone (P<0.05). Analysis of overall effect of different concentrations of MA alone, compared with combination of MA plus verapamil by mean infection rate of amastigotes in each macrophage showed a significant difference (P<0.05).These findings indicated some degree of synergistic effects between MA and verapamil on in vitro susceptibility of L. tropica to MA. Further works are required to evaluate this synergistic effect on animal model or volunteer human subjects

Indoor environment assessment of special wards of educational hospitals for the detection of fungal contamination sources: A multi-center study (2019-2021)

Background and Purpose: The hospital environment was reported as a real habitat for different microorganisms, especially mold fungi. On the other hand, these opportunistic fungi were considered hospital-acquired mold infections in patients with weak immune status. Therefore, this multi-center study aimed to evaluate 23 hospitals in 18 provinces of Iran for fungal contamination sources.Materials and Methods: In total, 43 opened Petri plates and 213 surface samples were collected throughout different wards of 23 hospitals. All collected samples were inoculated into Sabouraud Dextrose Agar containing Chloramphenicol (SC), and the plates were then incubated at 27-30ºC for 7-14 days.Results: A total of 210 fungal colonies from equipment (162, 77.1%) and air (48,22.9%) were identified. The most predominant isolated genus was Aspergillus (47.5%),followed by Rhizopus (14.2%), Mucor (11.7%), and Cladosporium (9.2%). Aspergillus(39.5%), Cladosporium (16.6%), as well as Penicillium and Sterile hyphae (10.4% each), were the most isolates from the air samples. Moreover, intensive care units (38.5%) and operating rooms (21.9%) had the highest number of isolated fungal colonies. Out of 256 collected samples from equipment and air, 163 (63.7%) were positive for fungal growth.The rate of fungal contamination in instrument and air samples was 128/213 (60.1%) and 35/43 (81.2%), respectively. Among the isolated species of Aspergillus, A. flavus complex (38/96, 39.6%), A. niger complex (31/96, 32.3%), and A. fumigatus complex (15/96, 15.6%) were the commonest species.Conclusion: According to our findings, in addition to air, equipment and instrument should be considered among the significant sources of fungal contamination in the indoor environment of hospitals. Airborne fungi, Hospital, Indoor air, Equipment, Sources of fungal contamination in the indoor environment of hospitals

Genska varijabilnost citrinin-toksikogenih sojeva Penicillium citrinum kao izvor opasnosti za zdravlje profesionalno izloženih populacija u sjevernom Iranu

We evaluated the ability of randomly amplified polymorphic DNA (RAPD) to type citrinin-producing Penicillium citrinum (P. citrinum) strains recovered from the forest’s air in northern Iran. A total of 12 P. citrinum strains (P1-P12) were characterised by citrinin production and random amplification of polymorphic DNA (RAPD) technique. All the strains produced citrinin with levels ranging from 1.5 μg mL-1 to 39.6 μg mL-1 (average value: 12.68 μg mL-1). Of 11 primers tested, eight primers produced polymorphic amplification patterns. These primers generated a total of 105 reproducible RAPD bands, averaging to 13.1 bands per primer. Dendrogram for each primer indicating the distance of the strains to each other was constructed. RAPD results showed that the collected strains constituted four different clusters. The first cluster included two isolates (P1 and P3). The second cluster included seven isolates (P2, P4, P5, P6, P7, P8, and P10). The third and fourth clusters included one isolate (P9) and two isolates (P11 and P12), respectively. We concluded that RAPD analysis might be used in providing genotypic characters for toxigenic P. citrinum strains typing in epidemiological investigations and public health related risk assessment.Polymorphic DNA) za tipiziranje sojeva Penicillium citrinum (P. citrinum) koji tvore citrinin, izoliranih iz šumskog zraka u području sjevernog Irana. Na osnovi tvorbe citrinina i primjenom RAPD-tehnike karakterizirano je ukupno 12 sojeva P. citrinum (P1-P12). Svi sojevi tvorili su citrinin u koncentracijama od 1,5 μg mL-1 do 39,6 μg mL-1 (srednja vrijednost: 12,68 μg mL-1). Od 11 testiranih početnica njih 8 dalo je polimorfne uzorke umnožavanja. Te početnice generirale su ukupno 105 reproducibilnih RAPD-vrpca, prosječno 13,1 vrpcu po početnici. Na osnovi podataka za svaku je početnicu konstruiran dendrogram iz kojega su vidljive međusobne udaljenosti sojeva. Rezultati dobiveni primjenom RAPD-tehnike pokazuju da se prikupljeni sojevi dijele u četiri različita klastera. U prvom su klasteru dva izolata (P1 i P3). U drugom je klasteru sedam izolata (P2, P4, P5, P6, P7, P8 i P10). Treći klaster sadržava jedan izolat (P9), a četvrti njih dva (P11 i P12). Zaključili smo da se RAPD-analiza može primijeniti u istraživanju genotipskih značajki toksinogenih sojeva P. citrinum, epidemiološkim istraživanjima i procjeni rizika povezanog s javnim zdravstvom

Three cases of brain hydatidosis in North Khorasan, Iran

Abstract Cystic hydatidosis is a serious public health problem in Iran. Although cysts can develop in almost all organs and the brain cysts are very rare. Here, we present 3 confirmed cases of brain hydatidosis and the patients who underwent successful surgery. Pathological examinations demonstrated the presence of cystic hydatidosis

Review of Development of Live Vaccines against Leishmaniasis

Leishmaniasis is a serious public health problem in both tropical and temperate regions, caused by protozoan parasites of the genus Leishmania. Cutaneous leishmaniasis is the most common form of leishmaniasis worldwide. After recovery from the initial infection in most of the patients, a long-lasting natural immunity will be established. In individuals with HIV infection or in immune deficient patients, the more dangerous forms can occur. Despite many attempts, there is no efficient vaccine for leishmaniasis. The main concern for live-attenuated vaccines is the possibility of returning to the virulent form. Therefore, the safety is an important point in designing a successful vaccine. Nonvirulent parasites as vaccine candidates are achievable through gamma-irradiation, long-term culture, random mutations induced by chemical agents, and temperature-sensitive mutations. The type of change(s) in such parasites is not known well and drawbacks such as reversion to virulent forms was soon realized. Leishmania tarentolae with capacity of adaptation to mammalian system has a potential to be used as nonpathogenic vector in vaccine programs. Due to its nonpathogenic intrinsic property, it does not have the ability to replace with the pathogen form. Moreover, the main problems are associated with the production of live vaccines, including lyophilization, storage, standards, and quality control that must be considered. In this review, we focused on the importance of different approaches concerning the development of a live vaccine against leishmaniasis

Cats and <i>Toxoplasma gondii</i>: A systematic review and meta-analysis in Iran

Toxoplasma gondii is a cosmopolitan zoonotic intracellular coccidian of the phylum Apicomplexa infecting warm-blooded animals and human beings. This protozoan causes a significant public health problem in humans and imposes considerable economic losses and damages to husbandry industries. The final host, cats, accounts for all of these significant burdens. Hence the present study was designed to analyse and review the overall prevalence rate of T. gondii infection in cats in Iran for the first time. In the present study data collection (published and unpublished papers, abstracts of proceedings of national parasitology congresses and dissertations) was systematically undertaken on electronic databases including PubMed, Google Scholar, Ebsco, Science Direct, Scopus, Magiran, Irandoc, IranMedex and Scientific Information Database. A total of 21 studies from 1975 to 2013 reporting prevalence of Toxoplasma infection in cats from different areas in Iran met the eligibility criteria. The pooled proportion of toxoplasmosis using the random-effect model amongst cats was estimated at 33.6% (95% confidence interval [CI] 22.05–46.41). The prevalence rate of cat toxoplasmosis in various regions of Iran ranged from 1.2% to 89.2%. Firstly, this study establishes a crude prevalence rate of T. gondii infection in cats. Secondly, it discusses the role of significant risk factors including sex, age and being either household or stray cats, in the epidemiology of the disease. Furthermore, the current study determines gaps and drawbacks in the prior studies that are useful to keep in mind to assist in designing more accurate investigations in future

Antileishmanial Activity of Lavandula angustifolia and Rosmarinus Officinalis Essential Oils and Nano-emulsions on Leishmania major (MRHO/IR/75/ER)

Background: The aim of present study was to evaluate antileishmanial effects of Lavandula angustifolia (L. angustifolia) and Rosmarinus officinalis (R. officinalis) medicinal plants essential oils and nano-emulsions on Leishmania major (L. major). Methods: The present study was performed in Leishmaniasis Reference Lab at Mazandaran University of Medical Sciences, Iran during 2016-2017. The IC50 values were calculated in both the promastigote and amastigote stages in J774 macrophage in comparison with meglumine antimoniate (MA) as positive control. In addition, cytotoxicity effects of essential oils and nano-emulsions prepared from both plants against macrophages were evaluated. Results: Both essential oil and nano-emulsion of Lavander and Rosemary showed anti-leishmania activity on promastigote with IC50=0.11 μl/mL, IC50=0.26 μl/mL, and IC50=0.08 μl/mL respectively. Moreover, during amastigote assay, Lavander and Rosemary essential oils and nano-emulsion were effective at least in concentration of 0.12 μl/mL and 0.06 µl/mL (P=0.0001) respectively, on mean infected macrophages (MIR) and amastigotes in macrophages (P=0.0001). Additionally, cytotoxicity assay against macrophage revealed no toxicity on the host cells at IC50 concentrations. Conclusion: The nano-emulsions of both plants were more effective than essential oil in both MIR and amastigote. However, in comparison with MA, the Lavander essential oil is more effective in reducing MIR. Rosemary nano-emulsion reduced MIR significantly more than MA in concentration of 0.25 μl/mL (P<0.001). Further investigations are recommended to evaluate the effect of these medicinal plants in murine models