The Larval Stages of Echinostoma spp. in Freshwater Snails as the First and Second Intermediate Hosts in Gilan and Mazandaran Provinces, Northern Iran.

Background: Identification of the larval stages of Echinostoma spp. in freshwater snails is an essential guide to continue monitoring the possibility of their transmission and the potential of echinostomiasis in areas where trematodes are the primary agent of parasitic diseases. The aim of this study was investigate Echinostoma using morphological and molecular techniques. Methods: The study was conducted in Gilan and Mazandaran Provinces, northern Iran, from April 2019 to October 2021. Overall, 5300 freshwater snails were randomly collected and were identified using external shell morphology. Meanwhile, snails infected with trematodes were studied via shedding and dissecting methods. Larvae stages of Echinostoma were identified and the genomic DNA of the samples was extracted. The PCR amplification of the ITSI gene was carried out for 17 isolates and products were sequenced. Seven sequences were deposited in GenBank. Results: Totally, 3.5% of snails containing three species (Stagnicola sp., Radix sp. and Planorbis sp.) were infected with two types of cercaria, E. revolutum with 37 and Echinostoma sp. with 45 spines in the collar. Moreover, 35% of the snails were infected with Echinostoma spp. metacercaria. Phylogenetic analysis illustrated that isolates were included in two ITSI haplogroups. Conclusion: Results showed the potential hazard of a zoonotic parasite as Echinostoma in northern Iran. The potential of disease environmental relationship investigation and resource control optimization is necessary for effective disease prevention and health management.

Gelatin Zymography of Major Proteases in Exosomes of Leishmania major Promastigotes.

Background: Enzymatic digestion of extra cellular matrix proteins by proteinases of Leishmania promastigotes is a complex process. Hence, studies on functional proteomics of these enzymes can help select these enzymes as possible vaccine candidates or selecting candidates for chemotherapy and immunotherapy. Several proteolytic enzymes are involved in virulence of Leishmania spp. These enzymes are mostly serine, cysteine and metalloproteases. We aimed to detect proteases in Leishmania promastigote exosomes. Methods: Serine, cysteine and metalloproteases were investigated in exosomes and lysate of L. major promastigote using gelatin zymography. The study was carried out in the Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, in 2021. Results: Zymography findings of metalloproteinases showed transparent bands, including a 63-kDa glycoprotein (GP63). This glycoprotein is a major surface metalloproteinase. In addition, transparent bands belonged to serin proteases and cathepsin were demonstrated in gels associated to Leishmania promastigote lysate and exosomes. Conclusion: Several metalloproteases, serin proteases and cathepsins were shown in promastigote lysate and exosomes of L. major, which could purified and used as fractions for immunodiagnostic.

Isolation and characterization of bacteriophages from wastewater sources on Enterococcus spp. isolated from clinical samples.

BACKGROUND AND OBJECTIVES: In recent decades, enterococcal resistance to antimicrobials has greatly increased. Furthermore, these chemicals include several side effects on the patients. Since no reports are available of the bacteriophages' effects on eukaryotic cells, they can be good solutions for multidrug-resistant bacterial problems. Therefore, the major aim of this study was to isolate bacteriophages from wastewaters on clinical antibiotic-resistant enterococci. MATERIALS AND METHODS: Clinical bacteria were isolated, then enterococcal isolates were identified using different methods. The antibiotic resistance scheme of the enterococcal isolates was assessed. The bacterial isolates were exposed to wastewater samples containing potential bacteriophages. Technically, isolated bacteriophages were studied by electron microscopy. RESULTS: Isolated bacteria were verified as Enterococcus faecium. Results showed that bacteriophages could easily be isolated from wastewater sources. The isolated bacteriophages were effective on E. faecium as well as Streptococcus dysgalactiae. Furthermore, these bacteriophages were challenged with five other bacteria (ATCC) with no visible effects. In general, the isolated bacteriophages belonged to the Myoviridae, Siphoviridae, and Inoviridae families. CONCLUSION: Further studies on bacteriophages and their efficacy on enterococcal strains could increase the treatment possibility of enterococcal infections. Due to these bacteriophages' effects on Streptococcus strains, bacteriophages may be used to treat streptococcal infections as well.

Designing of RNA Molecule Translating for Activitable Melittin as Selective Targeting of Leishmania Infected Cells.

BACKGROUND: Leishmaniasis is characterized by strong inflammatory responses with high levels of inflammatory cytokines that induce microRNA 21 and matrix metalloproteinases. Melittin has inhibitory effects on proliferation of various cells via induction of apoptosis. Melittin can be integrated in cell membranes and induce apoptosis. Thus, designation of biomolecules for the selective destroy of the infected cells is a treatment option. One approach is the precise engineering of constructs for the selective expression of melittin in the infected cells. METHODS: For this aim we designed a construct composing melittin nucleotide sequence and nucleotide sequence coding for polyanionic peptide function inhibitory element to further guarantee the selective function of melittin in inflamed tissues and infected cells, were included in a construct as melittin inhibitor via matrix metalloproteinase degradable linker. RESULTS: Reverse complementary sequences were designed so melittin sequences for the selective targeting of Leishmania could be expressed in infected cells using cell microRNA machinery. CONCLUSION: Translation machinery in infected cells with increased miR-21 could translate melittin, MMP linker and polyanionic inhibitor through a non-canonical pathway. Then, the MMP linker is degraded and selective killing of Leishmania infected cells would happen.

Optimization of antimicrobial efficiency of silver nanoparticles against three oral microorganisms in irreversible hydrocolloid impressions.

BACKGROUND AND OBJECTIVES: Silver nanoparticles (Ag-NPs) are potent antimicrobial agents, which have recently been used in dentistry. The aim of the current study was to optimize antimicrobial activity of Ag-NPs used in preparing irreversible hydrocolloid impressions against three microorganisms of Escherichia coli, Streptococcus mutans and Candida albicans. MATERIALS AND METHODS: After assessing antimicrobial activity of the compound using disk diffusion method, three parameters of concentration of Ag-NPs (250-1000 ppm), ratio of hydrocolloid impression material powder to water (0.30-0.50) and time of mixing (20.0-60.0 s), affecting antimicrobial activity of irreversible hydrocolloid impression materials against the three microorganisms, were optimized. This combined process was successfully modeled and optimized using Box-Behnken design with response surface methodology (RSM). Decreases in colony number of E. coli, S. mutans and C. albicans were proposed as responses. RESULTS: Qualitative antimicrobial assessments respectively showed average zone of inhibition (ZOI) of 3.7 mm for E. coli, 3.5 mm for S. mutans and 4 mm for C. albicans. For all responses, when the mixing duration and powder-to-water ratio increased, the circumstances (mixing duration of 59.38 s, powder-to-water ratio of 0.4 and Ag-NP concentration of 992 response) increased. Results showed that in optimum ppm, the proportion of decreases in colony numbers was maximum (89.03% for E. coli, 87.08% for S. mutans and 74.54% for C. albicans). Regression analysis illustrated a good fit of the experimental data to the predicted model as high correlation coefficients validated that the predicted model was well fitted with data. Values of R2Adj with R2Pred were associated to the accuracy of this model in all responses. CONCLUSION: Disinfection efficiency dramatically increased with increasing of Ag-NP concentration, powder-to-water ratio and mixing time.

Virulence determinants and biofilm formation of Acinetobacter baumannii isolated from hospitalized patients.

INTRODUCTION: Acinetobacter baumannii are nosocomial bacteria that are responsible for outbreaks and severe infections in hospitalized patients globally. The major target of this study was the characterization of virulence determinants and biofilm formation of A. baumannii isolates from hospitalized patients. METHODS: In total, 100 A. baumannii were collected from three hospitals in Tehran, Iran, 2017-2018. The isolates were assessed using phenotypic and genotypic methods and then screened for virulence factor encoding genes such as plcN and lasB using conventional polymerase chain reaction. Furthermore, bacterial biofilm formation, motility and hemolytic and proteolytic activities were assessed. RESULTS: Of 100 A. baumannii isolates, 20 isolates included plcN and four isolates included lasB using PCR assay. Overall, 21 isolates were negative for biofilm formation while 45, 20 and 14 of the total isolates were reported as weak, moderate and strong biofilm producers, respectively. All isolates were positive for bap genes using PCR. Moreover, 35 isolates were motile on Luria-Bertani media, 47 isolates were α-hemolytic on Brucella blood agar media and all isolates displayed proteolytic activity. CONCLUSIONS: Healthcare-associated infections with A. baumannii are a major concern, importantly due to their potency to acquire virulence factor genes. Therefore, shedding light in the discovery of new antimicrobial and/or therapeutic agents against virulent A. baumannii strains seem to be necessary.

A case report of typical leishmaniasis in dog.

Leishmania spp. are vector-borne flagellates transferred by sand flies. They cause cutaneous, mucocutaneous and visceral infections in mammals, especially in humans and dogs. A mature male boxer with ulcerative nodules around his eyes and snout was referred to Small Animal Hospital, Faculty of Veterinary Medicine, University of Tehran. Multiple cutaneous lesions were seen in physical examination. Mild leukocytosis, neutrophilia, left shift, lymphopenia, and thrombocytopenia were reported by the laboratory. Diagnosis was confirmed by the observation of amastigotes in blood samples and inside tissue macrophages. The infection was treated using pentavalent antimonial drug for four weeks.

The effect of melanocortin (Mc3 and Mc4) antagonists on serotonin-induced food and water intake of broiler cockerels.

The current study was designed to examine the effects of intracerebroventricular injections of SHU9119 [a nonselective melanocortin receptor (McR) antagonist] and MCL0020 (a selective McR antagonist) on the serotonin-induced eating and drinking responses of broiler cockerels deprived of food for 24 h (FD24). For Experiment 1, the chickens were intracerebroventricularly injected with 2.5, 5, and 10 µg serotonin. In Experiment 2, the chickens received 2 nmol SHU9119 before being injected with 10 µg serotonin. For Experiment 3, the chickens were given 10 µg serotonin after receiving 2 nmol MCL0020, and the level of food and water intake was determined 3 h post-injection. Results of this study showed that serotonin decreased food intake but increased water intake among the FD24 broiler cockerels and that these effects occurred in a dose-dependent manner. The inhibitory effect of serotonin on food intake was significantly attenuated by pretreatment with SHU9119 and MCL0020. However, the stimulatory effect of serotonin on water intake was not altered by this pretreatment. These results suggest that serotonin hypophagia and hyperdipsia were mediated by different mechanisms in the central nervous system, and that serotonin required downstream activation of McRs to promote hypophagia but not hyperdipsia in the FD24 chickens.

Antimicrobial Activity of Artemisia absinthium Against Surgical Wounds Infected by Staphylococcus aureus in a Rat Model.

The wound infection is one of the frequent complications in patients undergoing surgical operations. Staphylococcus aureus is the most common cause of surgical wounds. Artemisia absinthium has been shown to bear strong antimicrobial activity, especially against Gram-positive pathogens. This study was designed to investigate the antimicrobial effects of A. absinthium against surgical wounds infected by S. aureus in a rat model. Twenty male Sprague-Dawley rats were divided randomly into two equal groups of treated and control rats. A circular incision was created on the dorsal inter-scapular region of each rat. After skin wounding, rats were inoculated locally with 1 × 10(4) CFU of S. aureus at sites of skin wounds. The extract was applied topically twice a day throughout the experiment. Animals of the control group were left untreated. Results have revealed that topical application of A. absinthium extract on the infected wound sites produced significant antibacterial activity against S. aureus.

A survey on hydatidosis in livestock in Northern Iran based on data collected from slaughterhouses from 2004 to 2008.

Data have been collected from slaughterhouses in three provinces across the Northern Iran (Gilan, Mazandaran and Golestan) from March 2004 to March 2008. These data were collected to evaluate the prevalence of hydatidosis in slaughtered cattle, sheep and goats. During the study, 3,347,797 animals were slaughtered. These included 621,686 cattle, 1,719,725 sheep and 1,006,386 goats. The prevalence of infection in cattle, sheep and goats was 12%, 14.6% and 10.1%, respectively. The association of condemnation rates with seasons was not proven statistically.

Antimicrobial and heavy metal resistance in commensal enterococci isolated from pigs.

Antibiotic resistance in animal isolates of enterococci is of public health concern because of the risk of transfer of antibiotic resistance isolates or resistance determinants to consumers via the food chain. In this study, phenotypic and genotypic resistance in 192 pig isolates of enterococci to ampicillin, avilamycin, avoparcin, bacitracin, flavophospholipol, gentamicin, narasin, tetracycline, tiamulin, tylosin, vancomycin, virginiamycin, copper and zinc were investigated by susceptibility test and molecular methods. Resistance rates varied between the species but all isolates were susceptible to ampicillin, avilamycin, avoparcin, gentamicin and narasin but resistant to tetracycline and tylosin and intermediately resistant to copper. Only Enterococcus gallinarum and Enterococcus casseliflavus were resistant to vancomycin and virginiamycin resistance was present in less than half the Enterococcus faecium isolates. Zinc resistance was largely confined to Enterococcus faecalis but bacitracin resistance was uncommon in E. faecalis in comparison with the other species. Tiamulin resistance was common in all species except E. casseliflavus. Resistance to flavophospholipol was detected in most E. faecium isolates and in a high proportion of E. gallinarum, E. casseliflavus and E. hirae/durans but was only found in one isolate of E. faecalis. No tetO, rplC, rplD, vanA, vanB, vatA and vatD genes were found. The presence of ermB, tetL, tetM, tcrB, aac6-aph2, tetK, tetS, vanC1, vanC2, lsaA, lsaB and vatE varied between the species and largely corresponded to the susceptibility phenotype. The findings show that resistance to antibiotics of high clinical significance for nosocomial Enterococcus infections is absent, whereas antimicrobial resistance was detected for some other antibiotics including bacitracin, flavophospholipol, tetracycline, tiamulin, tylosin and virginiamycin.

Whole-genome sequencing and gene mapping of a newly isolated lytic enterococcal bacteriophage EFRM31.

Bacteriophages contribute greatly to bacterial evolution. There has been limited investigation of enterococcal bacteriophages, and only two enterococcal bacteriophages have been sequenced completely. In this study, a novel enterococcal bacteriophage, EFRM31, was isolated from a piggery effluent sample and then characterized. The complete bacteriophage genome was determined by shotgun sequencing. EFRM31 belongs to the family Siphoviridae (order Caudovirales) and has a circular double-stranded DNA genome. The putative EFRM31 genome consists of 16945 nucleotides with a low GC content (34.5%) and does not contain CpG islands. The EFRM31 genome contains 82 putative open reading frames, including 17 with identities to genes required for the assembly of a head-tail bacteriophage and 6 hypothetical proteins of unknown function. In general, the sequencing results from EFRM31 revealed considerable similarity to another enterococcal bacteriophage, EFAP-1. This identity and the order of shared genes suggest a close relationship or a common ancestor for these two bacteriophages.