Identification and genotyping of Echinococcus granulosus from human clinical samples in Guilan province, north of Iran.

Cystic echinococcosis (CE) is a significant health problem in both human and veterinary medicine. It is caused by the tapeworm Echinococcus granulosus (E. granulosus). The objective of this study was to investigate molecular diversity of E. granulosus from the paraffin-embedded human (FFPE) tissue samples using sequencing of mitochondrial genes. Thirty-five FFPE tissue samples were collected from different regions of Guilan province, north of Iran. Demographic data were recorded using a questionnaire. Five sections (1 mm) of the tissue were prepared and deparaffined using xylene and ethanol methods. Molecular analysis was performed using the Nad1 and Cox1 genes using PCR and DNA sequencing. Totally, 25 cases (71.43%) were women and 10 cases (28.57%) were men. The most affected age group was 21-30 yr old. The most of cysts were isolated from the liver (n = 19; 54.29%) and others in the lung (n = 16; 45.71%). The Cox1 and Nad1 genes were successfully amplified in 16 (45.71%) and 12 (34.28%) DNA samples from FFPE tissue. Sequencing analysis revealed that all samples were E. granulosus sensu stricto complex (G1 and G3). In this study, E. granulosus sensu stricto complex G1 and G3 were identified in human hydatid cysts and showed the presence of sheep/dog cycle in human infection. This finding confirmed and completed previous studies on the geospatial distribution of E. granulosus sensu stricto complex G1 and G3 in the southern and coastal areas of the Caspian Sea region.

In vitro anti-Acanthamoeba activity of the commercial chitosan and nano-chitosan against pathogenic Acanthamoeba genotype T4.

Acanthamoeba keratitis (AK) is a rare but serious infection of the eye and can lead to blindness. The effective and safe medical therapy remains unclear for AK until present. Antimicrobial activity and biological characteristic of chitosan encourage screening of it against Acanthamoeba. Thus, in vitro anti-amoebic activities of commercial chitosan and nano-chitosan were tested on pathogenic Acanthamoeba genotype T4, a causative agent of human AK. The Acanthamoeba spp. was isolated from the keratitis patient. The Acanthamoeba genotype T4 was approved using PCR method followed by sequencing technique. Chitosan nanoparticles was prepared using ionic gelation method and characterized by their physicochemical properties. In the present study, the in vitro activity of serial dilutions (12.5, 25, 50, 100, and 200 µL/mL) of commercial chitosan and nano-chitosan were evaluated against Acanthamoeba trophozoites and cysts. The finding of nano-chitosan particle size by DLS was 118 nm with a PDI of about 0.134. Zeta potential value was found to be 42.7 mV. The obtained results showed that the tested chitosan and nano-chitosan presented anti-amoebic activities dependent to time and concentration. The inhibitory effect of the chitosan and nano-chitosan is enhanced by increasing the concentration and incubation time. The inhibitory effect of nano-chitosan on both trophozoites and cyst was more than chitosan. According to the results, nano-chitosan shows the potent activity against Acanthamoeba T4 and could be used for the development of novel and safe therapeutic approaches in the future.

Morphological and Molecular Identification of Emerged Lophomonas blattarum Infection in Mazandaran Province, Northern Iran: First Registry-Based Study.

BACKGROUND: In the last decade, several cases of bronchopulmonary lophomoniasis (BPL) have been recorded. Little information is available about epidemiological aspects on Lophomonas infection among BPL patients. The present study was aimed to investigate the prevalence of Lophomonas spp. infection in patients who were referred to the Iranian National Registry Center for Lophomoniasis (INRCL), using morphological and molecular tests. SUBJECTS AND METHODS: We examined patients enrolled in the INRCL from 2017 to 2019 at the Mazandaran University of Medical Sciences, northern Iran. All bronchoalveolar lavage fluid (BALF) and two nasal discharges of the patients were examined by both microscopic and small-subunit ribosomal RNA (SSU rRNA) PCR methods. To confirm the species of Lophomonas, two positive samples were sequenced. RESULTS: In this study, 321 specimens (including 319 BALF and 2 nasal discharges) were microscopically examined. Lophomonas spp. was found in 45(14%) (n = 44 BAL; n = 1 nasal discharge). The mean age of infected patients was 54.9 ± 17.1 years. The following morphological characteristics were observed in both fresh and Papanicolaou-stained smears to identify Lophomonas spp. All microscopically positive specimens were confirmed with genus-specific PCR technique. The obtained sequences were deposited in Gen Bank under the accession numbers (MN243135-36). The BLAST analysis of our two sequences with the only available sequence in the Gen Bank of the Thailand strain of L. blattarum, showed identity of 99-100% and 98.51%, respectively. CONCLUSION: To the best of our knowledge, this is the first registry-based study regarding lophomoniasis worldwide. According to our study, the conventional PCR test is an available and reliable tool for confirming the Lophomonas parasite in clinical samples. Moreover, the results confirmed that L. blattarum is circulating at least in our region.

First Evidence for Colonizing of Acanthamoeba T4 Genotype in Urinary Tracts of Patients with Recurrent Urinary Tract Infections.

BACKGROUND: Limited evidence about the presence of Acanthamoeba spp. in urine specimens collected from urinary catheters of the patients in the intensive care units persuaded our study. No evidence has been found about colonizing of Acanthamoeba spp., in urinary tracts of patients with recurrent urinary tract infections (UTIs) yet. METHODS: In this study, 50 urine samples were collected from patients presenting with recurrent UTI. The type of bacteria causing UTI was determined by using bacteriological tests. To cultivate Acanthamoeba spp., in a sterile condition, 10 mL of urine was centrifuged and the sediment was cultivated on non-nutrient agar. Genotypes were determined by sequencing the DF3 region of the 18S rRNA gene. RESULTS: The bacteriological test findings on the urine samples of the UTI patients (n = 30) demonstrated that those were found to be positive for Escherichia coli (n = 17), Staphylococcus aureus (n = 6), Pseudomonas aeruginosa (n = 4) and Klebsiella spp. (n = 3) respectively. Moreover, a total of 50 urine samples was examined; 6 (6/50; 12%) and 11 (11/50; 22%) were positive by using culture and the PCR test for Acanthamoeba spp., respectively. Sequencing analysis showed all isolates were T4 genotype. CONCLUSIONS: Our data showed that the high relative prevalence of Acanthamoeba T4 genotype spp., in the urine of recurrent UTI patients. As well as, providing the first evidence for colonizing of the Acanthamoeba in the urinary tracts of patients with recurrent UTIs. These findings, warrant further investigation among those patients to fully appraise the role of Acanthamoeba spp., as possible latent carriers for resistant bacteria and biofilm formation in the future.

Toxoplasmosis among cancer patients undergoing chemotherapy: a population study based on the serological, molecular and epidemiological aspects.

BACKGROUND: Toxoplasmosis is highly prevalent in northern Iran and immunocompromised individuals are more susceptible to this infection. The present study aimed to determine the seroprevalence, parasitism and genetic diversity of Toxoplasma gondii among patients with cancer undergoing chemotherapy in northern Iran. METHODS: A total of 350 serum samples obtained from cancer patients were collected from laboratory centers in northern Iran. Immunodiagnosis and DNA detection were accomplished by ELISA and PCR. Thereafter, multiplex-nested PCR-restriction fragment length polymorphism was used for the genotyping of T. gondii. RESULTS: In general, out of 350 patients, 264 (75.4%) and 9 (2.57%) cases were positive for anti-T. gondii IgG and IgM, respectively. Moreover, 19 (5.43%) samples contained T. gondii DNA. From 19 positive samples, 10 high-quality samples with sharp and non-smear bands were selected to determine the genotypes of T. gondii. Accordingly, the samples were classified as genotype #1 (type II clonal; n=4, 40%), genotype #2 (type III clonal; n=3, 30%), genotype #10 (type I clonal; n=2, 20%) and genotype #27 (type I variant; n=1, 10%). CONCLUSIONS: As evidenced by the results, due to the high prevalence of T. gondii, cancer patients in northern Iran are at serious risk of severe toxoplasmosis and its complications. Therefore, oncologists need to regard this critical health problem as a matter requiring urgent attention.

The Mixed AII and BIII Genotypes of Human Giardia lamblia Isolate Circulating in Mazandaran Province, Northern Iran.

OBJECTIVES: Giardiasis is one of the human parasitic diseases by a flagellate protozoan named Giardia lamblia. This parasite is one of the most common organisms causing diarrhea in humans and a common gastrointestinal parasite in vertebrates. The purpose of this investigation was to determine the molecular epidemiology of Giardia lamblia (G. lamblia) by PCR-RFLP method in the Mazandaran Province and to investigate the possible relationship between parasite genotype and clinical symptoms. METHODS: In this study, 50 stool samples were collected from human cases of G. lamblia. referred to health centers in Sari, Babol, Nowshahr cities. The samples were examined by formalin-ether concentration procedure. DNA extraction was carried out on samples containing adequate Giardia cysts, and then PCR-RFLP was done on glutamate dehydrogenase (gdh) marker. RESULTS: In these patients, the most common signs were abdominal pain (42.9%), and diarrhea (47.6%); and least marked common sign was malaise (4.7%). Of all 21 isolates, 9 samples (42.8%) were found as genotype mixed AII and BIII, 6 samples (28.6%) belonged to genotype BIII and 6 (28.6%) were of genotype AII. Additionally, there was a significant correlation between AII genotype and clinical symptoms (P <0.05). According to the statistical tests, there was a significant relationship between abdominal pain and the intensity of infection (P<0.05). CONCLUSION: However, genotype AII and mixed AII and BIII genotypes were more common, respectively. The relatively high prevalence of mixed infection of these genotypes indicated the potential of zoonotic source of giardiasis among patients in Mazandaran Province.

Medicinal Plants with anti-Acanthamoeba Activity: A Systematic Review.

BACKGROUND: Recently, herbal medicine has received much attention in the literature. Several essential oils or plant extracts have been found to have anti-Acanthamoeba properties against trophozoites and cysts of Acanthamoeba spp. OBJECTIVE: The aim of this systematic review is to introduce anti-Acanthamoeba properties of some essential oils or plant extracts; perhaps the results of this research will be used to prevent and treat infectious diseases. METHODS: All published papers in English and Persian databases were systematically searched for some specific keywords to find articles that have influenced plant compounds on Acanthamoeba up to April 2018. Articles related to the subject were selected and studied. RESULTS: A total of 51 articles including 136 experiments (128 in vitro and 8 in vivo) between 1999 and 2018, met our eligibility criteria. Totally, 110 species of plants belonging to 34 families, mainly Lamiaceae (15 plant species) and Asteraceae (13 plant species) were studied against trophozoites and cysts of Acanthamoba in in vitro and in vivo. Most of the plant's species were Citrus (6 species), Allium (5 species), Peucedanum, Piper, Lippia, and Olive (4 species). The most frequently used parts were leaves, aerial parts, flowers, bark, rhizomes, and seeds. CONCLUSIONS: Recent studies have shown that many natural compounds have high anti-parasitic properties and low toxicity. Our research team hopes that the information provided in present systematic review can improve new experimental and clinical trials and herbal combination therapy. Further studies are needed to understand the molecular mechanisms in the anti-amoebic reactions of plant species and ocular toxicity of extracts in animal models.

Global status of synchronizing Leishmania RNA virus in Leishmania parasites: A systematic review with meta-analysis.

Leishmaniasis is one of the most neglected tropical diseases caused by protozoan parasites belonging to the genus Leishmania. There is much evidence regarding prevalence of Leishmania RNAvirus (LRV) causing Old World leishmaniasis (OWL) and New World leishmaniasis (NWL); however, a combined evidence-based knowledge on this topic is not still available. The purpose of this systematic review and meta-analysis was to address the global status of synchronizing LRV in Leishmania in the available literature. The data were systematically collected from the English electronic databases up to May 2018. Then, the studies were screened based on the inclusion and exclusion criteria. The random-effect model was used by forest plot with 95% confidence interval (CI). Overall, 877 samples from 17 articles were included in this study. Given species of Leishmania, the highest prevalence of LRV belonged to Leishmania (L.) Viannia (V.) guyanensis and L. V. braziliensis. Additionally, the virus was detected also in L. V. amazonensis, L. V. panamanensis, L. V. lainsoni, L. aethiopica, L. major and L. infantum. By random-effect model, the global prevalence of LRV was estimated to be 26.2% (95% CI: 14.4% - 40.1%). The high prevalence of LRV among causative agents of NWLisolated from the metastatic clinical forms suggests potential association of LRV with metastatic clinical forms in New World endemic regions. A comprehensive investigation on experimental and clinical aspects of LRV is needed to fully appraise the role of these viruses in pathogenicity of Leishmania parasites and their drug resistance.

A systematic review and meta-analysis of the genetic characterization of human echinococcosis in Iran, an endemic country.

Human echinococcosis is an infectious disease caused by tapeworms belonging to the species Echinococcus. This parasite has a worldwide distribution and is considered a neglected tropical disease by the World Health Organization. Due to the diversity of Echinococcus spp. hosts, as well as variation in geographical, climatic, and socio-ethnic conditions, the question of the strains or genotypes of Echinococcus spp. that are involved in human infections is important. The aim of this study was to provide a summary of the available data on genotypes of Echinococcus obtained from the Iranian population. Four international databases (PubMed, Scopus, Science Direct, and Web of Science) and 4 Persian databases (Magiran, Scientific Information Database, Iran Medex, and IranDoc) were searched for cross-sectional studies that reported the genotypes of Echinococcus spp. in human echinococcosis cases using molecular methods in Iran through July 2018. The Newcastle-Ottawa Scale was used to assess the quality of the selected studies. A total of 559 cases of human cystic echinococcosis were reported in the 21 included articles. The majority of cases belonged to genotype G1 (89.2%; 95% confidence interval [CI], 80.1 to 95.8), genotype G6 (8.2%; 95% CI, 2.8 to 15.9), and genotype G3 (2.3%; 95% CI, 1.1 to 3.9). Since genotype G1 of Echinococcus appears to be the most prevalent genotype affecting humans in Iran, disease control initiatives aimed at sheep intermediate hosts may be the most beneficial. In addition, educational programs and serological screening in individuals may help reduce the national impact of the disease.

Seroprevalence of Toxoplasma gondii infection in cancer patients: A systematic review and meta-analysis.

Toxoplasmosis, caused by Toxoplasma gondii, is a great public health concern in cancer patients, which can induce serious pathological effects. This systematic review and meta-analysis was performed to evaluate the worldwide seroprevalence rate of T. gondii infection among cancer patients. A search was conducted on five electronic databases that reported data on T. gondii seroprevalence in cancer patients. The searching process resulted in the inclusion of 57 studies. The results showed that T. gondii had the pooled prevalence of 30.8% in cancer patients using a random-effect model (95% CI: 26.3-35.6). Cancer patients had a higher overall prevalence of T. gondii infection, compared to those without cancer. Furthermore, the odds ratio of toxoplasmosis in cancer patients was 3.1 times, compared to that of controls (95% CI: 2.5-3.8, P < 0.0001). Toxoplasmosis had a higher prevalence in females (40%) than in males (33%). Furthermore, the age group of upper 40 years had the highest prevalence infection rate (30%). In addition, a significant association was also observed between toxoplasmosis infection and year (P < 0.001), type of cancer (P < 0.001), country (P < 0.001), gender (P < 0.001), age (P = 0.006) and diagnostic method (P < 0.001) in cancer patients. Considering the high prevalence of T. gondii infection in cancer patients and its serious outcomes, the researchers are suggested to carry out further studies to prevent and control toxoplasmosis among this population.

Effect of Matricaria chamomilla L. flower essential oil on the growth and ultrastructure of Aspergillus niger van Tieghem.

The antifungal activity of Matricaria chamomilla L. flower essential oil was evaluated against Aspergillus niger with the emphasis on the plant's mode of action at the electron microscopy level. A total of 21 compounds were identified in the plant oil using gas chromatography/mass spectrometry (GC/MS) accounting for 92.86% of the oil composition. The main compounds identified were alpha-bisabolol (56.86%), trans-trans-farnesol (15.64%), cis-beta-farnesene (7.12%), guaiazulene (4.24%), alpha-cubebene (2.69%), alpha-bisabolol oxide A (2.19%) and chamazulene (2.18%). In the bioassay, A. niger was cultured on Potato Dextrose Broth medium in 6-well microplates in the presence of serial two fold concentrations of plant oil (15.62 to 1000 microg/mL) for 96 h at 28 degrees C. Based on the results obtained, A. niger growth was inhibited dose dependently with a maximum of approximately 92.50% at the highest oil concentration. A marked retardation in conidial production by the fungus was noticed in relation to the inhibition of hyphal growth. The main changes of hyphae observed by transmission electron microscopy were disruption of cytoplasmic membranes and intracellular organelles, detachment of plasma membrane from the cell wall, cytoplasm depletion, and complete disorganization of hyphal compartments. In scanning electron microscopy, swelling and deformation of hyphal tips, formation of short branches, and collapse of entire hyphae were the major changes observed. Morphological alterations might be due to the effect on cell permeability through direct interaction of M. chamomilla essential oil with the fungal plasma membrane. These findings indicate the potential of M. chamomilla L. essential oil in preventing fungal contamination and subsequent deterioration of stored food and other susceptible materials.